detection of bovine lymphotropic herpesvirus dna in tissues of a bovine aborted fetus in québec...
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![Page 1: Detection of bovine lymphotropic herpesvirus DNA in tissues of a bovine aborted fetus in Québec Donald Tremblay, Ossama Allam, Richard Drolet and Carl](https://reader035.vdocument.in/reader035/viewer/2022081515/56649d785503460f94a5a155/html5/thumbnails/1.jpg)
Detection of bovine lymphotropic herpesvirus DNA in tissues of a bovine aborted fetus in Québec
Donald Tremblay, Ossama Allam, Richard Drolet and Carl A. Gagnon
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Project premises
In late 2008, the Molecular Biology Diagnostic Laboratory (MBDL) of the Faculty of veterinary medicine of the University of Montreal started a research project to investigate the etiological agents involved in cow abortion in Québec, Canada
-Molecular diagnostic (PCR)
4 principal pathogens:-BVDV-Bovine herpesvirus type 1 (IBR)-Leptospira spp-Neospora caninum
1 more (public health concern): Coxiella burnetti
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Data compilation
-Analyses are paid for. The results are given to the veterinarian as part of the regular diagnostic (24 hours delay after reception).
-The laboratory is sending coolers with identified collecting tubes to the submitting veterinary.
-vaccinal status of the herd-live attenuated, -killed viruses and -unvaccinated
-Historic-Blood for the serological follow-up
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Methodology and results
To date (May 2010)
-96 cases submitted
-86 placentae
-PCRs are done on
-placentae (when available)
-1 pool of all other tissues.
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Methodology and results
Standard qualitative PCR
IBR: polymerase A gene detection
Leptospira spp.:Gravekamp C. et al., 1993
Real-Time PCR (quantitative)
BVD: AgPath-ID BVDV (Life technologies)
Neospora caninum: Ghalmi F. et al., 2008
Coxiella burnetti: Klee S.R. et al., 2006
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Results
BVDV IBR LeptospiraNeosporacaninum
Coxiellaburnetti
5 (5,2) 6 (6,3) 2 (2,1) 9 (9,4) 1 (1,0)
Number of positive detected by PCR for each pathogen (%)
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Results
Since 1 case had BVDV and Leptospira spp.
-74 samples (77,1%) without any positive detection
-abortion rate in the herd?
-IBR: Slight increase since January 2010 (4 cases)
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Results
General bacterial detection by 16S rRNA sequencing:
-detection on placenta was inconclusive (environmental contamination of the samples)
-Very few amplification on individual tissues (contamination)
More target:
-Mycobacterium spp.
-Salmonella spp.
-Toxoplasma
-General Herpesvirus
-General Coronavirus
-Campylobacter spp.
-Listeria monocytogenes
-Chlamydophila spp.
-etc….
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General Herpesvirus
In January 2010
-case of BHV4 submitted to the MBDL
-First tested for IBR
detected with a pan-herpesvirus nested-PCR
Vandevanter et al., 1996
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General Herpesvirus
250
500
750
1000
1236
Exo I Exo II
Exo II’ Exo III
Motif A Motif B Motif C
DFA ILK TGV IYG KG1
- DNA polymerase gene- Final fragment of 210 bps- Degenerate primers- Product is unique for each viral species (with little strain variation)
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Results
- First 26 cases were tested for detection of pan-herpesvirus
- 2 amplifications:
- IBR (already reported case)- Bovine lymphotropic herpesvirus (BLHV)
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Bovine lymphotropic herpesvirus
- Herpesviridae family within the subfamily Gammaherpesvirinae
- First described in 1998 (Cornell University)- Rovnak J. et al 1998- Search of Herpesvirus as co-factor in the bovine
leukemia virus (BLV) pathogenesis- 80% positives in animals tested (n=147)
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- Report case in 2006 in the UK- Cobb et al., The Veterinary Report 2006- Cases of chronic unresponsive endometritis- 33% of a 180-head milking herd was affected with a purulent metritis that did not respond to standard antibiotic therapies
- Investigation in 2008- Banks et al., The Veterinary Journal 2008- small-scale investigation of UK dairy herds reporting recurrent
episodic or chronic non-responsive post-partum metritis- High prevalence in affected herds (but no control unaffected
herds)- No association with calving problems
Bovine lymphotropic herpesvirus
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Results
- Second consecutive abortion, fifth in the last year- Placenta and pool of tissues also positive for
Neospora caninum- Individual tissues of the aborted fetus tested
- Brain and lymph nodes were positive
- Tropism of Herpesvirus for neuronal cells and lymphocytes
- No histopathological lesion suggestive of a viral infection could be found in BLHV nested-PCR positives tissues
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Results
Investigation in the 60 other submitted placenta
- 6 others BLHV amplified
- 1/6 had Leptospira spp. infection
-Total of 7 cases (11,7% of 60 submitted placenta)
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Conclusion
- First description of a Bovine Lymphotropic Herpesvirus in Canada
- To our knowledge, it is the first time that BLHV is described in tissues of an aborted fetus
- It is difficult to ascertain that BLHV has a role to play in the cause of the abortion (presence of recognized pathogens and no significant viral lesions)
- Important to report the presence of BLHV in Canada because it is the first step toward the country improvement of bovine pathogens surveillance.
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Questions?