RESULTS
4 RESULTS
Majority of shrimp (Penaeus monodon) samples (healthy and moribund shrimps)
analyzed in this study were collected during the disease outbreaks Total numbers of
samples analyzed by PCR were 135 of which 67 were post larvae and rest were adult or
juvenile shrimp Of the 135 shrimp samples analyzed 49 (67) were positive for WSSV
(37 in single step PCR (Fig-1) and 30 in nested PCR (Fig-2)) MBV was detected in 29
(40) of the samples of which 21 were found positive in single step PCR (Fig-3) and 19
were positive in nested PCR (Fig-4) HPV was detected in 34 (47) of the samples (23
were found positive in first step PCR (Fig-5) and 24 in nested PCR (Fig-6) Table-41)
Presence of multiple viral infections was observed in many samples (Table-42)
Twenty-one samples (15) showed the presence of triple viral infections (WSSV MBV
and HPV) Dual viral infection with WSSV and HPV were seen in 17 (125) samples
Dual viral infection with WSSV and MBV were found in 6 and with MBV and HPV in 2
samples
41 Loose Shell Syndrome (LSS)
The LSS affected shrimps show hard or leathery shell but not soft and shrunken
tail meat with gap between the shell and the muscle tissue (causing the ldquoloose shellrdquo)
They also go off feed and may have swollen hind gut with whitish fluid The moribund
shrimp exhibited bacterial fungal and algal parasite infections on the exterior (Plate-1)
The infected shrimps move to the pond margins and finally die Studies conducted so far
have not conclusively determined the etiological agent for this syndrome Usually LSS
was noticed during the fourth month of the culture period In many of the farms the
water colour was dark-greenish (Plate-2) and in some farms LSS was also noticed in
normal bluish-grey colour ponds In some farms the pond bottom was degraded and
turned to black colour with rotten smell Affected shrimps were very lethargic and
reduced preening activity Some shrimps were affected so much that Zoothamnium sp
was densely covered the whole of the carapace and shrimp even lost its normal reflexive
activity when disturbed Some shrimps were discoloured to slight yellowish or greenish
due to algal growth
M 1 2 3 4 5
Fig 1 Agarose gel electrophoresis of WSSV one step PCR products Lane M 100bp
DNA ladder plus Lane 1 Positive control Lane 2-3 Samples positive for WSSV Lane
4 Sample negative for WSSV and Lane 5 Negative control
M 1 2 3 4 5
Fig 2 Agarose gel electrophoresis of WSSV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for WSSV Lane 4
Sample negative for WSSV and Lane 5 Negative control
M 1 2 3 4 5
Fig 3 Agarose gel electrophoresis of MBV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for MBV Lane 4
Sample negative for MBV and Lane 5 Negative control
M 1 2 3 4 5
Fig 4 Agarose gel electrophoresis of MBV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for MBV Lane 4
Sample negative for MBV and Lane 5 Negative control
M 1 2 3 4 5
Fig 5 Agarose gel electrophoresis of HPV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4
Sample negative for HPV and Lane 5 Negative control
1 2 3 4 5 6 M
Fig 6 Agarose gel electrophoresis of HPV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4-5
Sample negative for HPV and Lane 6 Negative control
Table-41 Results of shrimp samples from farms of South West and South East coast analyzed by PCR for WSSV MBV and HPV
WSSV MBV HPV No of Samples analyzed
I step II step I step II step I step II step
135 37(27) 30(22) 21(15) 19(14) 23(17) 24(17) Total 67(49) 40(29) 47(34)
Table-42 Presence of viruses in shrimp samples showing multiple viral infections
No of samples analyzed
WSSV+MBV+HPV WSSV+MBV WSSV+HPV MBV+HPV
135 21(~16) 6(4) 17(125) 2(1)
Plate-1 Loose shell syndrome shrimp
Plate-2 Loose shell syndrome affected pond Note the greenish colour of the pond water
The histopathological studies revealed extensive degenerative changes in
hepatopancreas including sloughing of the hepatopancreatic tubules from the basement
membrane (Plate-5) Necrotized areas showing deep basophilia karyorhexis and
pyknosis (Plate-6) were evident in hepatopancreas of most of the animals examined
Deeply basophilic inclusion-like bodies (Plate-7 and Plate-8) were observed in the cells
of tubular epithelium as well as in the inter-tubular space In some hepatopancreatic
tubules areas of focal haemocytic infiltration (Plate-9) with melanized nodular formation
with granulation tissues (Plate-10) were observed Atrophied tubules were characterized
by the absence of storage vacuoles However midgut traversing through hepatopancreas
appeared normal with intact epithelial cells to its basement membrane (Plate-11)
Histological sections also showed moderate to severe necrotic changes in the gills when
compared to normal gills (Plate-12) The changes were characterized by the necrosis of
pillar cells and detachment of cuticular layer from the underlying epithelium (Plate-13)
Many attached epicommensals were also observed in the gill filaments (Plate-13)
Integument appeared normal except the attachment of epicommensals in outer and inner
regions of carapace (Plate-15) in contrast to normal healthy shrimp (Plate-14) Moderate
degeneration of body muscles and heart tissue was also pronounced in all the animals
examined
Out of eleven LSS samples analyzed (Table-43) one sample showed multiple
viral infection with WSSV MBV HPV and LSNV (Fig-7 and Fig-8) Triple viral
infection was noticed in 2 samples one with WSSV MBV and HPV and another with
WSSV MBV and LSNV Five samples showed dual infection with WSSV and MBV and
only three were positive for HPV alone One sample was not infected with any of the
viruses tested except LSNV None of the samples tested for necrotizing hepatopancretitis
(NHP) bacterium by PCR was positive
Ten shrimp samples were collected from ponds showing the symptoms of loose
shell syndrome and subjected for bacteriology From each shrimp hepatopancreas and
hindgut were aseptically removed and streaked onto TCBS and TSAS plates Few
isolated colonies were subjected to biochemical analysis and the biochemical results
revealed the presence of V fluvialis V cincinnatiensis V fischeri V harveyii and V
natriegens
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
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1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
4 RESULTS
Majority of shrimp (Penaeus monodon) samples (healthy and moribund shrimps)
analyzed in this study were collected during the disease outbreaks Total numbers of
samples analyzed by PCR were 135 of which 67 were post larvae and rest were adult or
juvenile shrimp Of the 135 shrimp samples analyzed 49 (67) were positive for WSSV
(37 in single step PCR (Fig-1) and 30 in nested PCR (Fig-2)) MBV was detected in 29
(40) of the samples of which 21 were found positive in single step PCR (Fig-3) and 19
were positive in nested PCR (Fig-4) HPV was detected in 34 (47) of the samples (23
were found positive in first step PCR (Fig-5) and 24 in nested PCR (Fig-6) Table-41)
Presence of multiple viral infections was observed in many samples (Table-42)
Twenty-one samples (15) showed the presence of triple viral infections (WSSV MBV
and HPV) Dual viral infection with WSSV and HPV were seen in 17 (125) samples
Dual viral infection with WSSV and MBV were found in 6 and with MBV and HPV in 2
samples
41 Loose Shell Syndrome (LSS)
The LSS affected shrimps show hard or leathery shell but not soft and shrunken
tail meat with gap between the shell and the muscle tissue (causing the ldquoloose shellrdquo)
They also go off feed and may have swollen hind gut with whitish fluid The moribund
shrimp exhibited bacterial fungal and algal parasite infections on the exterior (Plate-1)
The infected shrimps move to the pond margins and finally die Studies conducted so far
have not conclusively determined the etiological agent for this syndrome Usually LSS
was noticed during the fourth month of the culture period In many of the farms the
water colour was dark-greenish (Plate-2) and in some farms LSS was also noticed in
normal bluish-grey colour ponds In some farms the pond bottom was degraded and
turned to black colour with rotten smell Affected shrimps were very lethargic and
reduced preening activity Some shrimps were affected so much that Zoothamnium sp
was densely covered the whole of the carapace and shrimp even lost its normal reflexive
activity when disturbed Some shrimps were discoloured to slight yellowish or greenish
due to algal growth
M 1 2 3 4 5
Fig 1 Agarose gel electrophoresis of WSSV one step PCR products Lane M 100bp
DNA ladder plus Lane 1 Positive control Lane 2-3 Samples positive for WSSV Lane
4 Sample negative for WSSV and Lane 5 Negative control
M 1 2 3 4 5
Fig 2 Agarose gel electrophoresis of WSSV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for WSSV Lane 4
Sample negative for WSSV and Lane 5 Negative control
M 1 2 3 4 5
Fig 3 Agarose gel electrophoresis of MBV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for MBV Lane 4
Sample negative for MBV and Lane 5 Negative control
M 1 2 3 4 5
Fig 4 Agarose gel electrophoresis of MBV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for MBV Lane 4
Sample negative for MBV and Lane 5 Negative control
M 1 2 3 4 5
Fig 5 Agarose gel electrophoresis of HPV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4
Sample negative for HPV and Lane 5 Negative control
1 2 3 4 5 6 M
Fig 6 Agarose gel electrophoresis of HPV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4-5
Sample negative for HPV and Lane 6 Negative control
Table-41 Results of shrimp samples from farms of South West and South East coast analyzed by PCR for WSSV MBV and HPV
WSSV MBV HPV No of Samples analyzed
I step II step I step II step I step II step
135 37(27) 30(22) 21(15) 19(14) 23(17) 24(17) Total 67(49) 40(29) 47(34)
Table-42 Presence of viruses in shrimp samples showing multiple viral infections
No of samples analyzed
WSSV+MBV+HPV WSSV+MBV WSSV+HPV MBV+HPV
135 21(~16) 6(4) 17(125) 2(1)
Plate-1 Loose shell syndrome shrimp
Plate-2 Loose shell syndrome affected pond Note the greenish colour of the pond water
The histopathological studies revealed extensive degenerative changes in
hepatopancreas including sloughing of the hepatopancreatic tubules from the basement
membrane (Plate-5) Necrotized areas showing deep basophilia karyorhexis and
pyknosis (Plate-6) were evident in hepatopancreas of most of the animals examined
Deeply basophilic inclusion-like bodies (Plate-7 and Plate-8) were observed in the cells
of tubular epithelium as well as in the inter-tubular space In some hepatopancreatic
tubules areas of focal haemocytic infiltration (Plate-9) with melanized nodular formation
with granulation tissues (Plate-10) were observed Atrophied tubules were characterized
by the absence of storage vacuoles However midgut traversing through hepatopancreas
appeared normal with intact epithelial cells to its basement membrane (Plate-11)
Histological sections also showed moderate to severe necrotic changes in the gills when
compared to normal gills (Plate-12) The changes were characterized by the necrosis of
pillar cells and detachment of cuticular layer from the underlying epithelium (Plate-13)
Many attached epicommensals were also observed in the gill filaments (Plate-13)
Integument appeared normal except the attachment of epicommensals in outer and inner
regions of carapace (Plate-15) in contrast to normal healthy shrimp (Plate-14) Moderate
degeneration of body muscles and heart tissue was also pronounced in all the animals
examined
Out of eleven LSS samples analyzed (Table-43) one sample showed multiple
viral infection with WSSV MBV HPV and LSNV (Fig-7 and Fig-8) Triple viral
infection was noticed in 2 samples one with WSSV MBV and HPV and another with
WSSV MBV and LSNV Five samples showed dual infection with WSSV and MBV and
only three were positive for HPV alone One sample was not infected with any of the
viruses tested except LSNV None of the samples tested for necrotizing hepatopancretitis
(NHP) bacterium by PCR was positive
Ten shrimp samples were collected from ponds showing the symptoms of loose
shell syndrome and subjected for bacteriology From each shrimp hepatopancreas and
hindgut were aseptically removed and streaked onto TCBS and TSAS plates Few
isolated colonies were subjected to biochemical analysis and the biochemical results
revealed the presence of V fluvialis V cincinnatiensis V fischeri V harveyii and V
natriegens
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
M 1 2 3 4 5
Fig 1 Agarose gel electrophoresis of WSSV one step PCR products Lane M 100bp
DNA ladder plus Lane 1 Positive control Lane 2-3 Samples positive for WSSV Lane
4 Sample negative for WSSV and Lane 5 Negative control
M 1 2 3 4 5
Fig 2 Agarose gel electrophoresis of WSSV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for WSSV Lane 4
Sample negative for WSSV and Lane 5 Negative control
M 1 2 3 4 5
Fig 3 Agarose gel electrophoresis of MBV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for MBV Lane 4
Sample negative for MBV and Lane 5 Negative control
M 1 2 3 4 5
Fig 4 Agarose gel electrophoresis of MBV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for MBV Lane 4
Sample negative for MBV and Lane 5 Negative control
M 1 2 3 4 5
Fig 5 Agarose gel electrophoresis of HPV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4
Sample negative for HPV and Lane 5 Negative control
1 2 3 4 5 6 M
Fig 6 Agarose gel electrophoresis of HPV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4-5
Sample negative for HPV and Lane 6 Negative control
Table-41 Results of shrimp samples from farms of South West and South East coast analyzed by PCR for WSSV MBV and HPV
WSSV MBV HPV No of Samples analyzed
I step II step I step II step I step II step
135 37(27) 30(22) 21(15) 19(14) 23(17) 24(17) Total 67(49) 40(29) 47(34)
Table-42 Presence of viruses in shrimp samples showing multiple viral infections
No of samples analyzed
WSSV+MBV+HPV WSSV+MBV WSSV+HPV MBV+HPV
135 21(~16) 6(4) 17(125) 2(1)
Plate-1 Loose shell syndrome shrimp
Plate-2 Loose shell syndrome affected pond Note the greenish colour of the pond water
The histopathological studies revealed extensive degenerative changes in
hepatopancreas including sloughing of the hepatopancreatic tubules from the basement
membrane (Plate-5) Necrotized areas showing deep basophilia karyorhexis and
pyknosis (Plate-6) were evident in hepatopancreas of most of the animals examined
Deeply basophilic inclusion-like bodies (Plate-7 and Plate-8) were observed in the cells
of tubular epithelium as well as in the inter-tubular space In some hepatopancreatic
tubules areas of focal haemocytic infiltration (Plate-9) with melanized nodular formation
with granulation tissues (Plate-10) were observed Atrophied tubules were characterized
by the absence of storage vacuoles However midgut traversing through hepatopancreas
appeared normal with intact epithelial cells to its basement membrane (Plate-11)
Histological sections also showed moderate to severe necrotic changes in the gills when
compared to normal gills (Plate-12) The changes were characterized by the necrosis of
pillar cells and detachment of cuticular layer from the underlying epithelium (Plate-13)
Many attached epicommensals were also observed in the gill filaments (Plate-13)
Integument appeared normal except the attachment of epicommensals in outer and inner
regions of carapace (Plate-15) in contrast to normal healthy shrimp (Plate-14) Moderate
degeneration of body muscles and heart tissue was also pronounced in all the animals
examined
Out of eleven LSS samples analyzed (Table-43) one sample showed multiple
viral infection with WSSV MBV HPV and LSNV (Fig-7 and Fig-8) Triple viral
infection was noticed in 2 samples one with WSSV MBV and HPV and another with
WSSV MBV and LSNV Five samples showed dual infection with WSSV and MBV and
only three were positive for HPV alone One sample was not infected with any of the
viruses tested except LSNV None of the samples tested for necrotizing hepatopancretitis
(NHP) bacterium by PCR was positive
Ten shrimp samples were collected from ponds showing the symptoms of loose
shell syndrome and subjected for bacteriology From each shrimp hepatopancreas and
hindgut were aseptically removed and streaked onto TCBS and TSAS plates Few
isolated colonies were subjected to biochemical analysis and the biochemical results
revealed the presence of V fluvialis V cincinnatiensis V fischeri V harveyii and V
natriegens
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Fig 3 Agarose gel electrophoresis of MBV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for MBV Lane 4
Sample negative for MBV and Lane 5 Negative control
M 1 2 3 4 5
Fig 4 Agarose gel electrophoresis of MBV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for MBV Lane 4
Sample negative for MBV and Lane 5 Negative control
M 1 2 3 4 5
Fig 5 Agarose gel electrophoresis of HPV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4
Sample negative for HPV and Lane 5 Negative control
1 2 3 4 5 6 M
Fig 6 Agarose gel electrophoresis of HPV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4-5
Sample negative for HPV and Lane 6 Negative control
Table-41 Results of shrimp samples from farms of South West and South East coast analyzed by PCR for WSSV MBV and HPV
WSSV MBV HPV No of Samples analyzed
I step II step I step II step I step II step
135 37(27) 30(22) 21(15) 19(14) 23(17) 24(17) Total 67(49) 40(29) 47(34)
Table-42 Presence of viruses in shrimp samples showing multiple viral infections
No of samples analyzed
WSSV+MBV+HPV WSSV+MBV WSSV+HPV MBV+HPV
135 21(~16) 6(4) 17(125) 2(1)
Plate-1 Loose shell syndrome shrimp
Plate-2 Loose shell syndrome affected pond Note the greenish colour of the pond water
The histopathological studies revealed extensive degenerative changes in
hepatopancreas including sloughing of the hepatopancreatic tubules from the basement
membrane (Plate-5) Necrotized areas showing deep basophilia karyorhexis and
pyknosis (Plate-6) were evident in hepatopancreas of most of the animals examined
Deeply basophilic inclusion-like bodies (Plate-7 and Plate-8) were observed in the cells
of tubular epithelium as well as in the inter-tubular space In some hepatopancreatic
tubules areas of focal haemocytic infiltration (Plate-9) with melanized nodular formation
with granulation tissues (Plate-10) were observed Atrophied tubules were characterized
by the absence of storage vacuoles However midgut traversing through hepatopancreas
appeared normal with intact epithelial cells to its basement membrane (Plate-11)
Histological sections also showed moderate to severe necrotic changes in the gills when
compared to normal gills (Plate-12) The changes were characterized by the necrosis of
pillar cells and detachment of cuticular layer from the underlying epithelium (Plate-13)
Many attached epicommensals were also observed in the gill filaments (Plate-13)
Integument appeared normal except the attachment of epicommensals in outer and inner
regions of carapace (Plate-15) in contrast to normal healthy shrimp (Plate-14) Moderate
degeneration of body muscles and heart tissue was also pronounced in all the animals
examined
Out of eleven LSS samples analyzed (Table-43) one sample showed multiple
viral infection with WSSV MBV HPV and LSNV (Fig-7 and Fig-8) Triple viral
infection was noticed in 2 samples one with WSSV MBV and HPV and another with
WSSV MBV and LSNV Five samples showed dual infection with WSSV and MBV and
only three were positive for HPV alone One sample was not infected with any of the
viruses tested except LSNV None of the samples tested for necrotizing hepatopancretitis
(NHP) bacterium by PCR was positive
Ten shrimp samples were collected from ponds showing the symptoms of loose
shell syndrome and subjected for bacteriology From each shrimp hepatopancreas and
hindgut were aseptically removed and streaked onto TCBS and TSAS plates Few
isolated colonies were subjected to biochemical analysis and the biochemical results
revealed the presence of V fluvialis V cincinnatiensis V fischeri V harveyii and V
natriegens
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
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PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
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1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Fig 5 Agarose gel electrophoresis of HPV one step PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4
Sample negative for HPV and Lane 5 Negative control
1 2 3 4 5 6 M
Fig 6 Agarose gel electrophoresis of HPV nested PCR products Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples positive for HPV Lane 4-5
Sample negative for HPV and Lane 6 Negative control
Table-41 Results of shrimp samples from farms of South West and South East coast analyzed by PCR for WSSV MBV and HPV
WSSV MBV HPV No of Samples analyzed
I step II step I step II step I step II step
135 37(27) 30(22) 21(15) 19(14) 23(17) 24(17) Total 67(49) 40(29) 47(34)
Table-42 Presence of viruses in shrimp samples showing multiple viral infections
No of samples analyzed
WSSV+MBV+HPV WSSV+MBV WSSV+HPV MBV+HPV
135 21(~16) 6(4) 17(125) 2(1)
Plate-1 Loose shell syndrome shrimp
Plate-2 Loose shell syndrome affected pond Note the greenish colour of the pond water
The histopathological studies revealed extensive degenerative changes in
hepatopancreas including sloughing of the hepatopancreatic tubules from the basement
membrane (Plate-5) Necrotized areas showing deep basophilia karyorhexis and
pyknosis (Plate-6) were evident in hepatopancreas of most of the animals examined
Deeply basophilic inclusion-like bodies (Plate-7 and Plate-8) were observed in the cells
of tubular epithelium as well as in the inter-tubular space In some hepatopancreatic
tubules areas of focal haemocytic infiltration (Plate-9) with melanized nodular formation
with granulation tissues (Plate-10) were observed Atrophied tubules were characterized
by the absence of storage vacuoles However midgut traversing through hepatopancreas
appeared normal with intact epithelial cells to its basement membrane (Plate-11)
Histological sections also showed moderate to severe necrotic changes in the gills when
compared to normal gills (Plate-12) The changes were characterized by the necrosis of
pillar cells and detachment of cuticular layer from the underlying epithelium (Plate-13)
Many attached epicommensals were also observed in the gill filaments (Plate-13)
Integument appeared normal except the attachment of epicommensals in outer and inner
regions of carapace (Plate-15) in contrast to normal healthy shrimp (Plate-14) Moderate
degeneration of body muscles and heart tissue was also pronounced in all the animals
examined
Out of eleven LSS samples analyzed (Table-43) one sample showed multiple
viral infection with WSSV MBV HPV and LSNV (Fig-7 and Fig-8) Triple viral
infection was noticed in 2 samples one with WSSV MBV and HPV and another with
WSSV MBV and LSNV Five samples showed dual infection with WSSV and MBV and
only three were positive for HPV alone One sample was not infected with any of the
viruses tested except LSNV None of the samples tested for necrotizing hepatopancretitis
(NHP) bacterium by PCR was positive
Ten shrimp samples were collected from ponds showing the symptoms of loose
shell syndrome and subjected for bacteriology From each shrimp hepatopancreas and
hindgut were aseptically removed and streaked onto TCBS and TSAS plates Few
isolated colonies were subjected to biochemical analysis and the biochemical results
revealed the presence of V fluvialis V cincinnatiensis V fischeri V harveyii and V
natriegens
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
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PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Table-41 Results of shrimp samples from farms of South West and South East coast analyzed by PCR for WSSV MBV and HPV
WSSV MBV HPV No of Samples analyzed
I step II step I step II step I step II step
135 37(27) 30(22) 21(15) 19(14) 23(17) 24(17) Total 67(49) 40(29) 47(34)
Table-42 Presence of viruses in shrimp samples showing multiple viral infections
No of samples analyzed
WSSV+MBV+HPV WSSV+MBV WSSV+HPV MBV+HPV
135 21(~16) 6(4) 17(125) 2(1)
Plate-1 Loose shell syndrome shrimp
Plate-2 Loose shell syndrome affected pond Note the greenish colour of the pond water
The histopathological studies revealed extensive degenerative changes in
hepatopancreas including sloughing of the hepatopancreatic tubules from the basement
membrane (Plate-5) Necrotized areas showing deep basophilia karyorhexis and
pyknosis (Plate-6) were evident in hepatopancreas of most of the animals examined
Deeply basophilic inclusion-like bodies (Plate-7 and Plate-8) were observed in the cells
of tubular epithelium as well as in the inter-tubular space In some hepatopancreatic
tubules areas of focal haemocytic infiltration (Plate-9) with melanized nodular formation
with granulation tissues (Plate-10) were observed Atrophied tubules were characterized
by the absence of storage vacuoles However midgut traversing through hepatopancreas
appeared normal with intact epithelial cells to its basement membrane (Plate-11)
Histological sections also showed moderate to severe necrotic changes in the gills when
compared to normal gills (Plate-12) The changes were characterized by the necrosis of
pillar cells and detachment of cuticular layer from the underlying epithelium (Plate-13)
Many attached epicommensals were also observed in the gill filaments (Plate-13)
Integument appeared normal except the attachment of epicommensals in outer and inner
regions of carapace (Plate-15) in contrast to normal healthy shrimp (Plate-14) Moderate
degeneration of body muscles and heart tissue was also pronounced in all the animals
examined
Out of eleven LSS samples analyzed (Table-43) one sample showed multiple
viral infection with WSSV MBV HPV and LSNV (Fig-7 and Fig-8) Triple viral
infection was noticed in 2 samples one with WSSV MBV and HPV and another with
WSSV MBV and LSNV Five samples showed dual infection with WSSV and MBV and
only three were positive for HPV alone One sample was not infected with any of the
viruses tested except LSNV None of the samples tested for necrotizing hepatopancretitis
(NHP) bacterium by PCR was positive
Ten shrimp samples were collected from ponds showing the symptoms of loose
shell syndrome and subjected for bacteriology From each shrimp hepatopancreas and
hindgut were aseptically removed and streaked onto TCBS and TSAS plates Few
isolated colonies were subjected to biochemical analysis and the biochemical results
revealed the presence of V fluvialis V cincinnatiensis V fischeri V harveyii and V
natriegens
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-1 Loose shell syndrome shrimp
Plate-2 Loose shell syndrome affected pond Note the greenish colour of the pond water
The histopathological studies revealed extensive degenerative changes in
hepatopancreas including sloughing of the hepatopancreatic tubules from the basement
membrane (Plate-5) Necrotized areas showing deep basophilia karyorhexis and
pyknosis (Plate-6) were evident in hepatopancreas of most of the animals examined
Deeply basophilic inclusion-like bodies (Plate-7 and Plate-8) were observed in the cells
of tubular epithelium as well as in the inter-tubular space In some hepatopancreatic
tubules areas of focal haemocytic infiltration (Plate-9) with melanized nodular formation
with granulation tissues (Plate-10) were observed Atrophied tubules were characterized
by the absence of storage vacuoles However midgut traversing through hepatopancreas
appeared normal with intact epithelial cells to its basement membrane (Plate-11)
Histological sections also showed moderate to severe necrotic changes in the gills when
compared to normal gills (Plate-12) The changes were characterized by the necrosis of
pillar cells and detachment of cuticular layer from the underlying epithelium (Plate-13)
Many attached epicommensals were also observed in the gill filaments (Plate-13)
Integument appeared normal except the attachment of epicommensals in outer and inner
regions of carapace (Plate-15) in contrast to normal healthy shrimp (Plate-14) Moderate
degeneration of body muscles and heart tissue was also pronounced in all the animals
examined
Out of eleven LSS samples analyzed (Table-43) one sample showed multiple
viral infection with WSSV MBV HPV and LSNV (Fig-7 and Fig-8) Triple viral
infection was noticed in 2 samples one with WSSV MBV and HPV and another with
WSSV MBV and LSNV Five samples showed dual infection with WSSV and MBV and
only three were positive for HPV alone One sample was not infected with any of the
viruses tested except LSNV None of the samples tested for necrotizing hepatopancretitis
(NHP) bacterium by PCR was positive
Ten shrimp samples were collected from ponds showing the symptoms of loose
shell syndrome and subjected for bacteriology From each shrimp hepatopancreas and
hindgut were aseptically removed and streaked onto TCBS and TSAS plates Few
isolated colonies were subjected to biochemical analysis and the biochemical results
revealed the presence of V fluvialis V cincinnatiensis V fischeri V harveyii and V
natriegens
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
The histopathological studies revealed extensive degenerative changes in
hepatopancreas including sloughing of the hepatopancreatic tubules from the basement
membrane (Plate-5) Necrotized areas showing deep basophilia karyorhexis and
pyknosis (Plate-6) were evident in hepatopancreas of most of the animals examined
Deeply basophilic inclusion-like bodies (Plate-7 and Plate-8) were observed in the cells
of tubular epithelium as well as in the inter-tubular space In some hepatopancreatic
tubules areas of focal haemocytic infiltration (Plate-9) with melanized nodular formation
with granulation tissues (Plate-10) were observed Atrophied tubules were characterized
by the absence of storage vacuoles However midgut traversing through hepatopancreas
appeared normal with intact epithelial cells to its basement membrane (Plate-11)
Histological sections also showed moderate to severe necrotic changes in the gills when
compared to normal gills (Plate-12) The changes were characterized by the necrosis of
pillar cells and detachment of cuticular layer from the underlying epithelium (Plate-13)
Many attached epicommensals were also observed in the gill filaments (Plate-13)
Integument appeared normal except the attachment of epicommensals in outer and inner
regions of carapace (Plate-15) in contrast to normal healthy shrimp (Plate-14) Moderate
degeneration of body muscles and heart tissue was also pronounced in all the animals
examined
Out of eleven LSS samples analyzed (Table-43) one sample showed multiple
viral infection with WSSV MBV HPV and LSNV (Fig-7 and Fig-8) Triple viral
infection was noticed in 2 samples one with WSSV MBV and HPV and another with
WSSV MBV and LSNV Five samples showed dual infection with WSSV and MBV and
only three were positive for HPV alone One sample was not infected with any of the
viruses tested except LSNV None of the samples tested for necrotizing hepatopancretitis
(NHP) bacterium by PCR was positive
Ten shrimp samples were collected from ponds showing the symptoms of loose
shell syndrome and subjected for bacteriology From each shrimp hepatopancreas and
hindgut were aseptically removed and streaked onto TCBS and TSAS plates Few
isolated colonies were subjected to biochemical analysis and the biochemical results
revealed the presence of V fluvialis V cincinnatiensis V fischeri V harveyii and V
natriegens
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-3 Histology of the Hepatopancreas of normal subadult shrimp Note the intact tubules (arrows) and different types of cells (B B-cells RR-cells)HampE400x
Plate-4 Histology of the Hepatopancreas of normal subadult shrimp Note the myoepithelial cells (arrowheads) haemocytes (arrow) EE-cells HampE1000x
E
B
R
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-5 Histology of the Hepatopancreas of LSS shrimp Note the detachment of the tubules from basement membrane (arrows) and other necrotic changes HampE400x
Plate-6 Histology of the Hepatopancreas of LSS shrimp Note the necrotized area showing the picnotic stage (arrow) of the nucleus HampE1000x
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-7 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-8 Histology of the Hepatopancreas of LSS shrimp Note deeply basophilic inclusion-like body (arrow) HampE1000x
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-9 Histology of the Hepatopancreas of LSS shrimp Note the degerative changes and accumulation of hemocytes HampE400x
Plate-10 Histology of the Hepatopancreas of LSS shrimp Note the granuloma formation HampE400x
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-11 Histology of the midgut of LSS shrimp Note the intact normal epithelial cells to its basement membrane HampE1000x
Plate-12 Histology of the gills of normal shrimp HampE400x
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-13 Histology of the gills of LSS shrimp Note the severe necrotic changes Arrow indicates Zoothamnium sp in gillsHampE 400x
Plate-14 Histology of integument of normal shrimp Note the different layers of the integument (arrow) with normal subcuticular epithelium (block arrow) and other underlying tissues HampE 400x
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-15 Histology of the integument of LSS shrimp Note the presence of epicommensals (arrows) in inner region of the carapace HampE400x
Plate-16 Histology of the hindgut of whitegut syndrome shrimp HampE100x
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Table-4 3 Presence of viruses in adult shrimp samples showing Loose shell syndrome
Sample WSSV MBV HPV LSNV NHP KLA-1 + + + + - KLA-2 + + + - - ALA-3 - - + - - ALA-4 - - + - - ALA-5 - - + - - KLA-6 - - - + - KLA-7 + + - + - KLA -8 + - - - - ALA-9 + - - - - ALA-10 + - - - - ALA-11 + - - - -
Total 7 3 5 3 0 Table-4 4 Results of the juvenile adult shrimp samples showing Swollen hind gut or white gut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KWA-1 + - + - KWA-2 - + - - KWA-3 + + - - AWA-4 + + + - AWA-5 + + - -
Total 4 4 2 0
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Fig 7 First step RT-PCR for detection of LSNV in Penaeus monodon M 100 bp DNA
ladder Plus (Gene Ruler TM genetix) Lane 1 Positive control Lane 2 Negative
control Lanes 3-5 Penaeus monodon samples with LSS positive for LSNV Lane 6
Normal adult shrimp sample Lane 7 Normal postlarvae of Penaeus monodon
Fig 8 Nested RT-PCR for detection of LSNV in P monodon Lane M 100bp DNA
ladder plus Lane 1 Positive control Lane 2-3 Samples negative for LSNV Lane 4-6
Sample positive for LSNV and Lane 7 Negative control
M 1 2 3 4 5 6 7
200 bp
M 1 2 3 4 5 6 7
150 bp
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
411 Slow growth syndrome
Three shrimps which were collected and preserved in RNA-friendly fixative
during the harvest of the crop were analyzed for LSNV WSSV MBV and HPV All
these shrimps were small in size (6-8g) when compared to other shrimps (25-30g) of the
same crop pond The culture period was 4-45 months After analyses the shrimps were
positive for only LSNV This was confirmed by sequencing the PCR products BLAST
(Altschul et al 1997) analysis of the sequences obtained in this study revealed 98
homology with the sequences of LSNV reported by Sritunyalucksana et al (2006) The
nucleotide sequence from this study was deposited in GenBank Accession Number
EF593037 (Fig-9) This is the first report of LSNV from India To characterize the new
virus (LSNV) closely related virusrsquos (Mushroom bacilliform virus) coat protein was
selected from the GenBank and three sets of primers were developed With these new
sets of primers RT-PCR was done using LSNV RNA as template After analysis there
were nonspecific bands in the gel Even after doing gradient PCR with annealing
temperatures ranging from 40ordm-60 ordmC we did not get any specific bands of 500bp 300bp
or 288bp which were expected from the new set of primers after amplification
42 Swollen Hindgut or White gut syndrome in adult shrimp
The shrimps having white gut syndrome were having bulged hindgut which can
be seen very clearly from outside The color of the hindgut was white or creamish when
compared to normal which was translucent Affected shrimps were of smaller in size and
there was no feed in the gut They were also found lethargic unlike healthy shrimps from
the same pond Pond bottom was turned to black in some of the farms where white gut
shrimps were collected But in some farms pond bottom was clean with normal color
Usually white gut syndrome was noticed during third month of the culture period Out of
five white gut syndrome-affected samples analyzed triple viral infection was seen in only
one sample whereas three samples showed dual viral infection (WSSV and HPV in one
and WSSV and MBV in two samples Table-44) MBV alone was noticed in only one
sample
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Fig 9 Nucleotide sequence of Laem-Singh virus
My NCBI[Sign In] [Register]
PubMed Nucleotide Protein Genome Structure PMC Taxonomy OMIM Books
Search CoreNucleotide for Go
You need JavaScript to work with this page
1 EF593037 Reports Penaeus monodon R[gi148710407] Links
bull Features bull Sequence
LOCUS EF593037 194 bp mRNA linear VRL 12-JUN-2007 DEFINITION Penaeus monodon RNA virus nonfunctional RNA-dependent RNA polymerase (RdRP) mRNA partial sequence ACCESSION EF593037 VERSION EF5930371 GI148710407 KEYWORDS SOURCE Penaeus monodon RNA virus ORGANISM Penaeus monodon RNA virus Viruses unclassified viruses REFERENCE 1 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Detection of Laem-Singh Virus in cultured Penaeus monodon from India JOURNAL Dis Aquat Org (2007) In press REFERENCE 2 (bases 1 to 194) AUTHORS PrakashaBK KarunasagarI and KarunasagarI TITLE Direct Submission JOURNAL Submitted (07-MAY-2007) Fishery Microbiology College of Fisheries Matsyanagar Mangalore Karnataka 575 002 India FEATURES LocationQualifiers source 1194 organism=Penaeus monodon RNA virus mol_type=mRNA isolation_source=hepatopancreas db_xref=taxon343515 country=India gene lt1gt194 gene=RdRP misc_feature lt1142 gene=RdRP note=nonfunctional RNA-dependent RNA polymerase due to mutation similar to GenBank Accession Number DQ127905 ORIGIN 1 ttgccttctc ccgagtggtc aggtttacgt gcaagagttc tcaggcttca tgaagtcagg
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
61 cattgtgata ctatctccac caattctcat gcccagatca tgctgcatat gcttgcttgc 121 aagcgctcgg gtgagcccgt gactcctata ttggcctgcg gtgatgacac tattcaagca 181 gctacctcag ccgg
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Bacteriological analysis of six shrimp samples collected from ponds showing the
symptoms of swollen hind gut or white gut revealed the presence of V fluvialis V
proteolyticus V carchariae V logei V metschnikovii and Bacillus sp When apparently
normal shrimp from the same culture pond was subjected to bacteriology only V
fluvialis was found to be present
The histopathological studies showed mild vacuolar degeneration of the tegmental
gland (Plate-16 17 18 19) associated with the hind gut when compared to normal shrimp
but the cuticular epithelium appeared normal (Plate-16 17 18) in the moribund shrimp
However hepatopancreas showed extensive degeneration wherein the complete tubular
architecture was lost in most of the tubules (Plate-20) especially the inner part of the
capsule
43 Swollen hindgut (SHG) syndrome in postlarvae
Of the 67 batches of post larval samples analyzed 14 were showing the symptoms
of swollen hind gut Post larvae with SHG showed enlargement and distention of the
hindgut folds and its junction with the midgut (Plate-22) although in some cases swelling
also occurred in the midgut of the sixth abdominal segment in contrast to healthy PL
(Plate-21) The abnormality caused cessation of the rhythmic movements of the hindgutndash
midgut junction resulting to failure of affected post-larvae to excrete fecal pellets
Hind gut appeared normal histologically in all the animals examined (Plate-25)
Some region of the midgut epithelium anterior to the junction of hindgut showed
degenerative changes (Plate-23 24) However severe necrotic changes in any of the cells
associated with the gut were not discernible in any of the animals examined Presence of
obvious bacterial colonies or parasitic infection was also not observed in the hind gut
region (Plate-25) Nevertheless the significant pathology observed was in
hepatopancreas where severely necrotized tubules were evident with occasional deeply
basophilic inclusion-like bodies (Plate-26) The degenerated tubule lumen also showed
aggregation of bacteria-like particles along with the tissue debris in few specimens
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-17 Histology of the hindgut of whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrow) HampE400x
Plate-18 Histology of hindgut in Whitegut syndrome shrimp Note normal chitin lining (arrows) and mild degeneration of the tegmental gland (block arrows) HampE 400x
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-19 Histology of hindgut in Whitegut syndrome shrimp Note mild degeneration of the tegmental gland (block arrow)HampE 1000x
Plate-20 Histology of the hepatopancreas of whitegut syndrome shrimp Note the severe necrosis of the tubules HampE100x
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-21 Sixth abdominal segment of normal postlarvae showing normal hindgut (arrow) 100x
Plate-22 Sixth abdominal segment of the postlarvae showing the swollen hindgut Arrow indicates the swollen hindgut 100X
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-23 Histology of Swollen hindgut syndrome in postlarvae HampE 100x
Plate-24 Histology of midgut region of Swollen hindgut syndrome in postlarvae Note the degenerative changes in midgut region anterior to the hindgut-midgut junction HampE 400x
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Plate-25 Histology of Swollen hindgut syndrome in postlarvae Note normal hind-gut with chitin lining (arrow) and absence of bacterial or parasitic infection HampE 1000x
Plate-26 Histology of Hepatopancreas in Swollen hindgut syndrome in post larvae Note the severely necrotized tubules (arrow) with deeply basophilic inclusion like-body (arrowheads) HampE 100x
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Out of 14 batches of post larvae (showing SHG) analyzed nine were positive for
WSSV and HPV each and only two for MBV (Table-45) Triple viral infections were not
noticed in any of the samples analyzed Dual viral infection with WSSV and HPV was
noticed in six post larval samples whereas dual viral infections with WSSV and MBV
were seen in only two samples Five samples showed infection with HPV alone
Bacteriological analysis of post larval samples showed the presence of V
alginolyticus
44 White Tail Disease (WTD) in fresh water prawn
Freshwater prawns (Macrobrachium rosenbergii) Artemia nauplii Acetes sp
were analyzed for Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus
(XSV) by RT-PCR Four berried prawns which were collected from Nethravati estuary
were analyzed by taking few eggs and pleopods separately and subjected to RT-PCR and
found negative for MrNV and XSV Two Artemia nauplii and Artemia flakes were also
found to be negative Six juvenile samples were collected from a farm in Hasson district
were also found negative for both MrNV and XSV Some prawn larvae which were
showing necrosis of tail muscle and abdomen were also found negative for both the
viruses Even some Acetes sp which were collected at the same farm in Nellore also gave
negative result Water sample collected from prawn farm in Hasson was also negative
45 Prevalence of WSSV MBV and HPV in post larvae Prevalence of viruses in post larval samples from West Coast of India was
analyzed by PCR A total of 505 samples were screened for WSSV MBV and HPV
(Table-46) Of these 25 (49) were positive for WSSV 83 (1643) were positive for
MBV and 55 (1089) for HPV In addition to single viral infection dual and triple viral
infections were observed Six samples showed infection with two viruses (WSSV and
MBV) Simultaneous infections with MBV and HPV were found in 11 samples The co-
occurrence of WSSV and HPV was also noticed in 9 samples (178) One sample was
found infected with WSSV MBV and HPV
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Table-4 5 Results of the post larval samples showing Swollen hindgut analyzed for WSSV MBV HPV and LSNV
Sample WSSV MBV HPV LSNV KSP-1 - - + - ASP-2 + - + - ASP-3 - - + - ASP-4 + - + - ASP-5 + - + - ASP-6 + - + - ASP-7 + - + - ASP-8 + - + - ASP-9 + - - - KSP-10 - - - - KSP-11 - - - - KSP-12 + + - - KSP-13 + + - - ASP-14 - - + - ASP-15 - - + - ASP-16 - - - - ASP-17 - + + - ASP-18 - - + -
Total 9 3 12 0
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Table-46 Results of prevalence of viruses in post larvae
Year 2004 2005 2006
Viruses
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
No of samples positive
Prevalence ()
WSSV 4 289 6 363 15 742 MBV 29 2101 27 1636 27 1336 HPV 28 2028 18 109 9 445 WSSV + MBV 1 07 0 0 5 247 MBV + HPV 4 289 7 424 0 0 WSSV + HPV 7 507 1 06 1 049 WSSV+MBV+HPV 0 0 1 06 0 0 No of samples analyzed
138 165 202
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively
Year-wise analysis of the samples showed that in the year 2004 out of 138
samples 4 (289) were positive for WSSV 29 (2101) for MBV and 28 (2028) for
HPV One (07) sample showed dual viral infection with WSSV and MBV
Simultaneous occurrence of MBV and HPV was observed in 4 (289) samples and
WSSV and HPV in 7 (507) samples
In the year 2005 out of 165 samples analyzed six (363) were positive for
WSSV 27 (1636) for MBV and 18 (109) for HPV Seven (424) samples showed
the simultaneous presence of MBV and HPV and one (06) was found to have WSSV
and HPV Triple viral infection (WSSV MBV amp HPV) was noticed in one sample In the
year 2006 of 202 samples analyzed 15 (742) were positive for WSSV 27 (1336)
for MBV and 9 (445) for HPV alone Five (247) samples showed dual viral
infection with WSSV and MBV only one sample for WSSV and HPV but none for MBV
and HPV
The year-wise analysis reveals that there is an increasing trend in the level of
WSSV infection year after year but decreasing trend in the level of individual infections
of MBV and HPV However there is fluctuation in the dual and triple viral infections
year after year Dual viral infection with WSSV and MBV was 07 in 2004 but
decreased to zero in the year 2005 and again increased to 247 in the year 2006 In the
case of dual viral infection with MBV and HPV there was an increase in the rate of
infection from 289 in 2004 to 424 in 2005 but decreased to zero in the year 2006
However there is decreasing trend in the case of dual viral infections with WSSV and
HPV from 507 in 2004 to 06 and 049 in 2005 and 2006 respectively