dost r-traifalgar feed seminar

7/21/2019 DOST R-Traifalgar Feed Seminar

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Alternative Feed Ingredient: “Water Hyacinth

Leaf Protein Concentrate as Feed Ingredient in

Shrimp Diet”.

Rex Ferdinand Traifalgar

Nicole Niña Chavez

Jenny Gonzaga

College of Fisheries and Ocean Sciences

University of Philippines in Visayas



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Executive Summary

Water hyacinth plant (Eichhornia crassipes , Mart Solms) is a fast growing and

invasive aquatic plant known to cause flooding of major waterways in the country. The

utility of this bioresource is low in the country but this resource has a high potential to be

developed as feed material to support the needs of the growing aquaculture industry.

The present studies were undertaken to develop a method of producing feed grade

leaf protein concentrate and test this feed material as a replacement for soybean meal

in the diets of cultured shrimp the black tiger shrimp (P.monodon ) and P. vannamei .

Findings of the study indicate that dietary replacement of soybean meal with water

hyacinth protein concentrate in the diet of the white shrimp P. vannamei supported

better growth of this species. Optimum dietary level of water hyacinth protein

concentrate that can replace soybean meal is about 25%. This dietary replacement

dose resulted to an FCR, SGR, PER that is similar to those obtained in groups fed with

diet having full soybean meal content. Results of the trial using P. monodon larvae show

that 75% of the soybean meal protein could be replaced by water hyacinth leaf protein

concentrate in the diet. These results indicate that P. monodon can tolerate a higher

level of this feed material in the diet.

The higher dietary replacement levels of soybean meal with water hyacinth protein

concentrate could be attributed to the balanced and complete amino acid composition of

this feed ingredient. Moreover, levels of heavy metals in the water hyacinth leaf protein

concentrate were found to be minimal and are considered as safe. Collectively these

findings elucidated that water hyacinth leaf protein concentrate could be used as a



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replacement for soybean meal in the practical diet of black tiger shrimp and the white

shrimp.

The Philippine aquaculture industry is a major provider of quality food products and an

important source of revenue and employment of coastal communities. However, the

profitability and sustainability of this industry is dependent on the input costs of

production with feed cost contributing the highest to the overall production costs.

Currently, the industry is dependent on imported feed ingredients including soybean

meal, fishmeal, and fish oil for feed production. Further, the costs of these key feed

components are rising due to the increasing global demands caused by usage

competition among the animal growing industries and is hypothesized to hamper the

growth and expansion of the industry.

. In commercially manufactured feeds for aquaculture, marine protein sources

including fish, shrimp and squid meals are the primary protein sources being included in

the feed at approximately 25% by weight (Tacon & Barg, 1998). Traditionally fishmeal is

the protein source used in aquaculture diets, but the current drop in global fishmeal

production due to overexploitation and rising demands led to the rise in cost making it

uneconomical to use as a sole ingredient for aquaculture feed use. Soybean meal is the

most important plant protein, which is currently used in combination with marine

proteins in feeds for cultured species. Studies evaluating soybean meal as replacement

Introduction



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for marine proteins in Litopenaeus vannamei diets have shown that up to 40% of a

marine protein mix (53% menhaden fishmeal, 34% shrimp waste meal, and 13% squid

meal) could be replaced by solvent-extracted soybean meal, without reducing the

growth of this shrimp (Lim and Dominy, 1990). A 25 to 30% dietary level of soybean

meal as a replacement for 40 to 50% of the marine animal protein in shrimp feeds

appears as to be optimal (Swick et al., 1995). In fish, specifically tilapia, soybean meal

can replace up to 25% of fish meal in diets when given in combination with blood meal.

Substitution in proper proportions does not significantly lower SGR and FCR but when

plant proteins are given solely, growth performance is significantly reduced and

mortalities are common at certain inclusion levels .

However, concerns regarding the safety of the use of genetically modified

soybean and the rising cost of this ingredient due to demand competition by the other

sectors of the animal growing industries are factors that drive the search for a cheaper

and locally produced plant protein ingredient that could replace soybean meal in the

diets of cultured aquatic animals.

The use of locally produced plant-derived feed ingredient as full or partial

substitute for soybean and fishmeal on aquaculture feed could be a potential approach

to lessen the country’s dependence on imported and costly feed inputs. Protein

concentrate meal derived from the photosynthetic proteins of green leaf could be a

potential a substitute for soybean meal in the formulated diets of cultured species.

Concentrated protein derived from aquatic plants has potential as aquaculture feed



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ingredient. Earlier scientific reports indicate that protein concentrate sourced from plant

materials are well utilized by animals including fish. Protein concentrates derived from

plant leaves have been tested as feed material for swine, cow, chicken, and fish (Telek

and Graham, 1983). Moreover, protein concentrates from rice bran, wheat, and corn

have been tested and proven a feasible protein source that could satisfy the protein

requirements of carp, turbot, and salmon. (Kaur and Saxena, 2005, Rónyai and Gál,

2005, Robaina et al ., 1999).

However, previous workers have focused on investigating protein concentrate

from terrestrial plants and works on developing and testing protein materials from

aquatic plants as feed are less investigated. Aquatic plants are known to have a fast

growth rate and could reach a high biomass in a short period of time. Utilization of this

biomass as feed resource is of high potential for aquaculture. The water hyacinth

Eichhornia crassipes is considered an ideal material for the development of a feed-

grade protein concentrate because it has a little or no direct utility value for human use,

grows rapidly in tropical environment, could be harvested in large quantities, and is

considered a nuisance species. In addition, this aquatic plant passes the criteria as an

ideal stock or source of protein concentrate for its raw protein content is above 20% of

the dry weight (Virabalin et.al, 1993). Although this plant contains a good amount of

protein in the leaf but the other component of the biomass like carbohydrate and fibers

limit the optimum utilization of this biomass as feed ingredient for aquatic species. The

production of a protein concentrate however eliminates the fibrous bulk component and

only protein and some carbohydrates could be obtained and is considered as ideal

feed ingredient.



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The project on its first year of implementation has able to developed protein

concentrate from water hyacinth leaves. The protein concentrate collected from fresh

water hyacinth ranges from 5-7 % of the dry biomass with protein content of 28-30%,

making this material as a potential feed ingredient for aquaculture use.

. The present investigation was designed to evaluate the replacement of soybean meal

with water hyacinth leaf protein concentrate in the diets of P. monodon and P.

vannamei. This work also elucidates the biological value of this feed material in the diet

of cultured shrimp.

In the present project activities, basic investigations on the nutritional value of

water hyacinth protein concentrate as a replacement of soybean meal component in the

diet of two species of cultured shrimp were investigated. Feeding trials were done and

biological growth performance, nutrient assimilation and biochemical composition of the

test animals were evaluated and used as indices to assess the feed value of the water

hyacinth leaf protein concentrate.

Experimental Animals & Rearing Facilities

Pathogen free P. vannamei postlarvae,(PL10, ten days after metamorphosis),

weighing 5.0 mg, and P. monodon (PL12, 5.0mg) were obtained from a commercial

Methodology

Experimentation/Feeding Trials



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hatchery in the province of Iloilo and transported to the facilities of the Institute of

Aquaculture Multi-Species Hatchery (University of the Philippines Visayas, Miagao,

Iloilo). Postlarvae were stocked and acclimated in 2-ton fiberglass tank indoors for 7

days. The postlarvae were fed artemia and commercial postlarvae feed three times

daily. Following the acclimation, the two hundred forty postlarvae of each species were

randomly collected and distributed in 24 tanks (30-L volume).Twelve tanks were used to

hold each species constituting the 4 experimental treatments in triplicate following a

completely randomized design (CRD). Tanks were supplied with constant aeration

maintaining oxygen near at saturation levels. Water temperature was at the range of

28°C to 30°C, at a salinity of 32 to 35 psu and the animals were exposed to normal

photoperiod.

For the fish trials, Nile tilapia (Tilapia nilotica ) and Milkfish (Chanos chanos ) fry

were obtained from the hatchery of Southeast Asian Fisheries Development Center-

Aquaculture Department (SEAFDEC-AQD), Tigbauan, Iloilo. The fry were transported to

the university hatchery facilities and were acclimatized to the laboratory conditions for

two weeks. The animals were maintained fed the control diet at 30% body weight, four

times daily (0800, 1100, 1400, and 1700 h).

Feeding trials were conducted in a recirculating culture system with 12 (95L)

tanks equipped with a with a biological filter, sedimentation tank, and sediment filter (5

µm). Continuous aeration was provided and water quality parameters were monitored

regularly. Water temperature and pH were measured twice daily (0800 and 1400 h)

using a laboratory mercury thermometer and hand-held digital pH meter, respectively.



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Normal photoperiod was used and levels of dissolved oxygen, total ammonia nitrogen,

and nitrite were measured using test kits (Advance Pharma Co., Ltd., Bangkok,

Thailand). Optimum levels of water temperature (27.11 ± 0.15oC), pH (8.87 ± 0.06),

dissolved oxygen (8.50 ± 0.27 ppm), total ammonia nitrogen (0.11 ± 0.03 ppm), and

nitrite (0.18 ± 0.05 ppm) were observed and maintained all throughout experimental

period.

Experimental Diets & Growth Trials

SHRIMP EXPERIMENT (P.monodon & P.vannamei )

Four experimental diets (Table 1) were formulated to evaluate the effects of

increasing protein substitution of soybean meal with water hyacinth leaf protein

concentrate (WHLPC) at 0%, 25%, 50%, and 75% designated as D1, D2, D3, and D4,

respectively. The control diet (D1) contained soybean meal, fishmeal and squid meal as

the main protein sources. All four diets were formulated to be isonitrogenous (40%

crude protein) and isolipidic (8% crude lipid). The feeding experiment was run in 12

aquaria that constitute the four dietary treatments in triplicate. Pelleted diets were

prepared by mixing of the dry ingredients with fish oil/soybean oil and added with

cooked starch forming dough. The formed dough was pelleted by passing through a

meat grinder twice and the resulting strands were oven-dried at 60°C overnight. After

drying, strands were broken, sieved to a convenient size, and stored at 4°C until use. At

the beginning of the experiment, 250-350µm crumbles were used, increasing its size up



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to 800-1000µm at the end of the experiment. This formulated diet was used for both

P.monodon and P.vannamei feeding trials.



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TABLE 1. Composition of experimental diets (g/100 g dry weight) used in feeding

trials with P. monodon and P. vannamei .

% Dietary protein replacement of soybean meal with WHLPC

D1 (0%) D2 (25%) D3 (50%) D4 (75%)

INGREDIENT

Soybean meal 30 22 14 8

WHLPC 0 18 35 48

Fish meal 20 20 20 20

Squid meal 14 14 14 14

Fish oil/soybean oil 5 4 3.5 3

Starch 20 14 8.98 2.48

Vitamin premix 2 2 2 2

Mineral premix 0.5 0.5 0.5 0.5

Lecithin 0.5 0.5 0.5 0.5

BHT 0.02 0.02 0.02 0.02

Lignobond 1.5 1.5 1.5 1.5

Alpha-cellulose 6.48 3.48 0 0

TOTAL 100 100 100 100

a Analyzed values from Oversea Feeds Corporation Quality Control Laboratory, Balud, San Fernando, Cebu;1D1, D2, D3, D4,

experimental diets in which soybean meal was replaced by WHLPC at 0, 25, 50, and 75% dietary protein basis; 2

Vitamin premix.

Β-carotene, 3.0 M.I.U. kg-1; cholecalciferol, 0.6 M.I.U. kg

-1; thiamin, 3.60 g kg

-1; riboflavin, 7.20 g kg

-1; pyridoxine, 6.60 g kg

-1;

cyanocobalamine, 0.02 g kg-1; α-tocopherol, 16.50 g kg

-1; menadione, 2.40 g kg

-1; niacin, 14.40 g kg

-1; pantothenic acid, 4.00 g kg

-1;

biotin, 0.02 g kg-1; folic acid, 1.20 g kg

-1; inositol, 30.00 g kg

-1; stay C, 100.00 g kg

-1.;

3Mineral premix. P, 12.0%; Ca, 12.0%; Mg,

1.5%; Fe, 0.15%; Zn, 0.42%; Cu, 0.21%; K, 7.50%; Co, 0.011%; Mn, 0.160%; Se, 0.001%; Mo, 0.0005%; al, 0.0025%; I, 0.04%.

Proximate Analysis of Shrimp Dieta

D1 (0%) D2 (25%) D3 (50%) D4 (75%)

Crude Protein 40.86 40.85 41.75 41.12

Crude Fat 9.11 8.74 8.89 8.83

Ash 10.20 10.66 11.13 12.27

Dry Matter 94.41 94.14 93.44 92.6



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Animals were fed three times daily (0900, 1300, and 1700) following a fixed

feeding regimen which was adjusted every ten days during sampling. Experimental

diets were given to the treatment groups for 70 days of culture for P. vanna mei and 60

days of culture for P. mo nodon. Uneaten feed, feces, molts, and dead shrimp in each

tank were collected and 30% of the water was replenished daily prior to feeding.

Normal photoperiod was observed and water temperature, salinity, dissolved oxygen

(DO), and pH were monitored daily. Total ammonia nitrogen (TAN), ammonia, and

nitrite-nitrogen were monitored and measured weekly using test kits (Aqua Am and

Aqua Nite). To assess growth rate, shrimp were bulk weighed per tank every ten days

throughout the trial. At the end of the feeding trial, fish samples from each tank were

pooled, weighed, and processed for proximate body composition analysis.

Biological performance of the test animals as a response to the test diets were

evaluated following these biometrics:

• Percent Weight Gain (% wt gain) = ((final body weight – initial body weight) /

initial body weight) x 100

• Specific Growth Rate (SGR) = ((ln average final weight – ln average initial

weight)/number of days) x 100

• Feed Conversion Efficiency (FCE) = weight gain (g) / dry feed intake (g)

• Protein Efficiency Ratio (PER) = weight gain (g) / protein intake (g)

• Survival (%) = (Final number of shrimp/ Initial number of shrimp) x 100

• Protein Retention = (Protein gain in fish (g) / Protein intake in food (g)) x 100

• Lipid Retention = (Lipid gain in fish (g) / Lipid intake in food (g)) x 100



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General Biochemical Analyses

Dietary protein sources including soybean meal and WHLPC were subjected to

proximate composition analysis prior to the formulation of the experimental diets to

ensure balanced nutrient levels in the test diets (Table 3). Samples were analyzed for

moisture by oven drying at 105 °C until dried. Total nitrogen (N) was determined by the

Kjeldhal method (Kjeltec system, Tecator, Sweden) and CP content calculated as

N*6.25. Lipid was determined using hexane extraction by the Soxhlet method.

TABLE 3. Proximate composition of soybean meal & water hyacinth leaf protein

concentrate.

Crude

Protein

Crude

Fat

Crude

Fiber

Moisture Ash NFE HC

insoubes

!H"PC 20#34 3#37 5#04 8#79 15#48 46#98 1#15

$o%bean

&ea

44#79 0#60 3#66 10#60 7#12 33#23 0#08

Ash analysis was determined by furnace ashing at 550-600°C. Crude fiber (CF)

content was determined according to AOAC (1990). In addition, WHLPC was analyzed

for presence of heavy metals (Table 4) by Flame Atomic Absorption Spectrophotometric

Method. The amino acid profile of the water hyacinth protein concentrate was also



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analyzed using HPLC amino acid analyzer (Table 5). The comparison of essential

amino acid composition of P.vannamei and water hyacinth is presented in Table 6.

TABLE 4. Heavy metal content of water hyacinth leaf and protein concentrate.

$a&'e () Para&etersa

*

Moisture

* Ash A+e# ,ota

Cad&iu&

&-./-

A+e# ,ota

Co''er

&-./-

A+e# ,ota

"ead

&-./-

!HP"b

8#03 0#08 13#90 0#04 0#20 0#02 5#25 0#14 0#39 0#03

!H"PC

6#10 0#15 11#60 0#11 0#31 0#02 22#09 0#88 0#44 0#06

Mai&u& i&itd

2#00 0#00 25#00 0#00 40 0#00

Analyzed values from Analytical Services Laboratory, Department of Chemistry, College of Arts and Sciences, University of the

Philippines Visayas, Miagao, Iloilo;bWater hyacinth powdered leaves;

cWater hyacinth leaf protein concentrate;

dMaximum levels set

for feedstuff as reported by FIN Fact Sheet.



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TABLE 5. Complete amino acid profile of water hyacinth protein concentrate.

AM(N AC() PF("E

-.100- 'rotein

E$$EN,(A"

Ar-inine 6#6

Histidine 2#2

(soeuine 5#5

"euine 9#6

"%sine 5#1

Phen%aanine 6#0

Methionine 1#3

,hreonine 5#3

,r%'to'han 1#4

aine 7#5

NNE$$EN,(A"

Aanine 6#5

As'arti aid 10#2

C%stine 0#4

%ine 6#5

uta&i aid 7#3

Proine 5#6

,%rosine 2#9

$erine 10#2



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TABLE 6. Comparative essential amino acid profiles of P.vannamei carcass and

water hyacinth protein concentrate.

aBased on the work of Forster et al., 2002;

aActual analyzed value.

Essentia A&ino AidsaEAA 'ro:ie; *

'rotein , P.

vannamei

-.100- 'rotein

bEAA 'ro:ie;

!H"PC

-.100-'rotein

Arginine 6.1 6.6

Histidine 1.6 2.2

Isoleucine 2.7 5.5

Leucine 4.7 9.6

Lysine 4.8 5.1

Methionine & Cystine 2.7 6.0

Phenylalanine & tryptophan 5.4 1.3

Threonine 2.5 5.3

Tryptophan 0.7 1.4

Valine 3.1 7.5



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Analysis of the protein concentrate showed that the feed material contains 20%

crude protein and the overall proximate composition analysis is presented in Table 3 .

This data indicate that this material could be used as a protein source and energy in

animal diets. The amino acid profile of WHLPC shows that this ingredient contains

almost all of the required dietary amino acids for a feed ingredient that are essential for

growth (Table 5). Moreover, the profile of WHLPC amino acid was found to be almost

similar to the P. vannamei carcass essential amino acid profile (Table 6) indicating that

this feed material could be an ideal feed protein for this organism. Analysis of the

presence of heavy metals in both the unprocessed dried leaf and the processed

WHLPC showed that cadmium, copper, and lead are present in both of these materials.

The process of concentrating protein from water hyacinth leaf tends to increase the

these metals in the protein concentrate, but it is worth emphasizing that the levels of

these metals in the protein concentrate are so low that it do not exceed the maximum

levels set for a feed material (Table 4).

Overall water quality parameters observed throughout the feeding trials were found

optimal for the needs of shrimp to attain growth and survival. In both trials using P.

monodon and P.vannamei mortality was minimal with no significant differences in

survival observed among the dietary treatments.

Weight gain of the test shrimp increased at every sampling period. At the end of the

feeding trial, weight gain values of P.vannamei fed diets containing the full soybean

meal were observed to be the highest among the treatments but did not differ

V. Results



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significantly with the treatment with 25% WHLPC replacement. Significantly lower

weight gain was observed in shrimp fed the highest level of replacement at 75%. In

contrast to P. vannamei , the numerical value of the weight gain of P. monodon at the

termination of feeding trial was found to be higher in treatments receiving the diets with

WHLPC as a replacement for soybean meal. Higher numerical value of weight gain was

observed in the highest replacement level of 75% however, statistical analysis indicates

that this value is not different to the other treatments including those that received the

full soybean meal in the diet.

TABLE 8. Proximate composition of shrimp (P.vannamei ) carcass (g/100g wet weight)

before and after the feeding trial.

Replacement levels

Initial 0% 25% 50% 75%

Protein 6.79±0.21 18.61±2.01 17.87±0.57 17.72±0.82 17.38±1.57

Lipid 4.11±0.48 7.72±1.47 8.42±1.84 6.19±1.58 5.61±0.78

Ash 3.41±0.24 14.34±0.81 12.57±0.14 13.27±0.29 11.51±5.89

Moisture 85.7±7.13 76.53±2.04 75.97±0.57 77.41±0.14 72.5±5.76

In trial with P.vannamei growth indices including specific growth rate( SGR),

protein efficiency ratio (PER), and the ability to convert eaten feeds to biomass , feed

conversion efficiency (FCE) was observed to be similar in treatments maintained with



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the 25% WHLPC replacement and the control treatment that received the full soybean

meal diet. In this trial, it was generally observed that higher WHLPC replacement levels

beyond the 25% elicited a significant reduction in shrimp growth performance in terms

of SGR, FCE, and PER. Similar pattern of growth responses were also observed in the

trial with P.monodon . In contrast to P. vannamei no significant decline in biological

performance in terms of feed conversion and growth rate as the level of WHLPC

replacement increases. Biological growth indices were similar in dietary treatments with

75% replacement level and those that received the full soybean meal diet as the control

group.

Dietary replacement level of soybean meal with WHLPC in P. vannamei has no

significant effects on the biochemical composition of the shrimp carcass. No significant

differences in terms of total lipid and protein contents were found among treatments

(Table 8). However, the ability of P. vannamei to convert feed protein and lipid into

tissue biomass as measured in terms of protein and lipid retention were significantly

decreased as the soybean replacement level increases. Highest protein retention was in

shrimp fed the control diet without (0%) WHLPC replacement but was not significantly

different from shrimp fed diet containing 25% WHLPC. In addition, the lipid retention

showed no significant difference between the control and the treatment with 25%

WHLPC replacement. Both protein and lipid retentions were reduced at increasing

WHLPC replacement in the diet (Table 11).



TABLE 11. Growth performance and nutrient utilization of P. vannamei fed diets with increas

WHLPC. Values are means ± SEM of three replicates. Numbers within the same column with d

significantly different (P < 0.05).

!H"PC )ietar%

Protein

e'ae&ent

A<!1

* !ei-ht ain2

$ur+i+a ate3

$4

PE5

FCE6

0% 1.91± 0.27a

38167.27± 5339.34a

81.67 ± 7.26

8.47 ± 0.20a

1.84 ± 0.12a

71.17 ± 4.65a

25% 1.46 ± 0.12ab

29104.73± 2362.87ab

88.33 ± 4.41 8.10 ± 0.11ab

2.09 ± 0.11a 80.78 ± 4.18

a

50% 1.11 ± 0.11c 22165.56± 2250.99

c 88.33 ± 7.26 7.71 ± 0.14 1.72 ± 0.02

a 67.38 ± 0.81

a

75% 0.68 ± 0.09c 13480± 1706.23

c 100 ± 0.00 6.99 ± 0.19

c 1.37 ± 0.01

b 52.40 ± 0.40

b



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Dietary soybean meal replacement with WHLPC in the diets of P. monodon

showed that the replacement levels has no significant influence on the biochemical

deposition of nutrients in the shrimp carcass as measured by protein and lipid retentions

(Table 14). In contrast to P. vannamei wherein protein and lipid retentions tends to

decline as the replacement levels increases , in P. monodon protein and lipid retentions

at the highest level of replacement at 75% were found not different from the control

group receiving the full soybean meal diets.



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TABLE 14. Biological response and nutrient retentions of P. monodon fed diets with

WHLPC as soybean meal replacement. Values within similar rows with different letter

superscripts are significantly different.

Soybean meal replacement levels (g/100g)

Biological

Indices

0 25 50 75

Survival (%) 77.78±1.9 85.56±5.09 93.333±6.67 86.667±5.77

Weight Gain

(%)

1881.76±485 1779.68±117 2423.53±503 1725.00±274

SGR 3.95±0.34 3.91±0.08 4.28±0.28 3.86±0.19

PER 0.38±0.06 0.35±0.10 0.48±0.09 0.35±0.05

FCE 20.93±5.50 18.92±12.8 21.84±10.37 18.73±12.19

Lipid Retention 0.38±0.1 0.36±0.02 0.74±0.15 0.60±0.09

Protein

Retention

7.1±1.8 6.04±0.37 9.04±1.8 6.62±1.09



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The present report, to our knowledge is the first time that water hyacinth leaf

protein concentrate (WHLPC) is used as a dietary protein source for the cultured

shrimp, P. vannamei and P. monodon . The feed material was observed to be

acceptable for both shrimp species used in the present study and increasing levels of

WHLPC, as a replacement for soybean meal, did not affect the overall survival of the

test shrimp species suggesting absence of toxic substances or contaminants that has

the potential to cause acute toxicological effects. Moreover the detected levels of heavy

metals that include, Lead, Cadmium and Copper in the WHLPC is at the allowable level

for a feed material further supporting the high potential of this material as a feed

ingredient.

Results of the feeding trials with P. vannamei indicate that 25% soybean protein

replacement was found optimum and is comparable to the diet with full soybean meal in

promoting normal biological performance in terms of weight gain, specific growth rate,

feed efficiency, protein efficiency ratio, and lipid and protein retentions. In P. monodon

trial about 75% of soybean protein can be replaced with WHLPC without negative

influence on the overall growth performance of this species.

Liang and Lovell (1971) were the first to test the use of water hyacinth as feed

ingredient in rations of channel catfish. Hyacinth protein extract was produced and

incorporated in increasing amounts in the diets with 40% as the highest inclusion level.

Optimum replacement level of water hyacinth protein extract that promoted biological

growth response similar to the control diet was at 5-10% of the diet. Results of the

Discussion



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present experiment indicate a higher replacement level of WHLPC in the diet that is

25% in terms of protein and is about 18% of the diet by weight. The differences in

values obtained in catfish and in L. vannamei in the present study could be due to the

difference in the metabolism of both fish and shrimp. In addition, L. vannamei is

considered to utilize a good amount of plant protein in the diet and catfish is a carnivore

that requires high amount of animal protein in the diet. In contrast, the high tolerance of

P. monodon to higher level of WHLPC in the diet may indicate that all the necessary

requirement of this shrimp in terms of energy and amino acid may have been satisfied

with the feed containing the highest content of WHLPC. The difference in response

between the two species of shrimp to replacement levels with WHLPC could possibly be

due to the differences in feeding behavior and metabolism of these species. However,

these aspects need further elucidation.

Previous studies in the inclusion of plant protein materials in the diets of aquatic

animals generally show that higher inclusion levels led to a significant decrease in

overall growth performance. Jackson et al.(1982) reported that tilapia fed Leucaena leaf

meal contributing 25% and 50% of the total protein in the diet exhibited depressed

growth. Findings from Ng and Wee (1989) also showed that the optimum replacement

levels of treated cassava leaf meal dietary protein without adversely affecting tilapia

growth and protein utilization was observed to be about 20%. Furthermore, Xie and

Jokumsen (1997) reported that increasing levels (0-51%) of potato protein concentrate

(PPC) in the diet of rainbow trout significantly decreased growth parameters with

specific growth rate decreasing linearly with increasing levels of dietary PPC. These



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earlier reports concur with the results of the present study in that overall growth indices

decline beyond 25% dietary replacement of the plant protein sources.

The overall lipid and protein retentions of P. monodon was not affected by high

level of WHLPC as a replacement for dietary soybean meal, suggesting that this feed

ingredient could support the required amino acid to attain optimum growth of this

species. On the other hand, the lipid and protein contents of P. vannamei exhibited

decreasing trend at the higher dietary soybean meal replacement levels. Shrimp fed

with the control diet (0% replacement) has higher protein and lipid content that is similar

with that of the treatment with 25% replacement level. Shrimp fed with the highest

content of WHLPC resulted to have lower carcass protein and fat contents. High levels

of dietary plant proteins in diets tend to increase daily fat gain and reduce nitrogen

retention but results of current experiment on nutrient retention showed that a

decreasing trend in protein and lipid retentions are observed with increasing levels of

WHLPC in the diet. In rainbow trout limitation of rice protein concentrate inclusion in the

diet is primarily due to imbalanced amino acid profile (Palmegiano et al.2006). The

imbalanced amino acid content in the diet usually results to a decrease carcass protein

with high fat content. This is in contrast to the findings of the present study suggesting

that the decrease in carcass protein and lipid contents could not be due to the amino

acid limitations. Amino acid analysis of the protein concentrate indicates a complete

amino acid profile and is closely similar to the amino acid profile of shrimp carcass

protein. The amino acid profile of the WHLPC is generally considered ideal as a feed

protein for L. vannamei .



25

Heavy metal content of the WHLPC indicates that this feed ingredient contained safe

levels of copper, lead, and cadmium. The growth inhibition at higher replacement level

in P. vannamei is very unlikely to be caused by the presence of these metals and or

caused by digestion problem. In-vitro digestibility shows that the WHLPC is more

digestible than the Danish fishmeal and soybean meal. The dominant protein molecule

in leaf protein concentrate is a protein related to photosynthesis known as the

carboxydismutase, ribulose 1, 5-biphosphate carboxylase or, more simply, fraction-I

protein. This protein is highly conserved in green plants and known to be highly

digestible with proteases. The high digestibility of this feed material may possibly be

related to inhibition of growth performance at higher inclusion levels observed in the

present study with P.vannamei . Amino acid influx in the absorbing intestinal enterocytes

is a condition known to limit the availability and absorption of low molecular weight

peptides and amino acids in feed. It is tempting to speculate that the high digestibility of

this feed ingredient can result to amino acid influx, limiting amino acid absorption thus

reducing the overall growth performance of the shrimp. Further, the low growth

performance of P. vannamei fed diets containing higher levels of WHLPC as protein

replacement could also be due to some anti-nutritional factors found in most plant-

based ingredients. Anti-nutrient is the major factor suggested to limit the inclusion level

of most plant protein sources in the diet of aquatic animals (Makkar 1993). Xie and

Jokumsen (1997) reported that the presence of heat-stable, glucoalkaloid solanine in

potato protein concentrate depressed growth in rainbow trout. Similar to the findings of

the present study, decreasing growth performance including weight gain, SGR, PER,

feed efficiency, and protein retention were observed. In addition, Jackson et al. (1982)



26

showed that condensed tannins limit the replacement level of copra in the diet of tilapia

fingerlings even at such low levels of replacement as 25%. The presence of anti-nutrient

compounds was not examined in WHLPC in the present study and needs a thorough

investigation in the future.

Collectively the present results indicate that Water Hyacinth Leaf Protein

Concentrate can replace 25% dietary protein of soybean meal in formulated diets for P.

vannamei and 75% in the diets of P. monodon. These replacement levels do not affect

the overall growth performance but affects the carcass composition of these cultured

species. Utilization of WHLPC in aquafeeds as an alternative protein source to soybean

meal may help to reduce the feed cost in aquaculture making it more sustainable and

competitive as an economic venture.



27

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dost r-traifalgar feed seminar

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