FISH MEAL REPLACEMENT IN PRACTICAL DIETS FOR PACIFIC WHITE
SHRIMP (Litopenaeus vannamei) REARED IN GREEN WATER SYSTEMS
Except where reference is made to the work of others, the work described in this
thesis is my own or was done in collaboration with my advisory
committee. This thesis not include proprietary
or classified information
______________________
Elkin A. Amaya
Certificate of Approval:
___________________________ ___________________________
William Daniels D. Allen Davis, Chair
Associate Professor Associate Professor
Fisheries and Allied Aquacultures Fisheries and Allied Aquacultures
___________________________ ___________________________
David B. Rouse Stephen L. McFarland
Professor Acting Dean
Fisheries and Allied Aquacultures Graduate School
FISH MEAL REPLACEMENT IN PRACTICAL DIETS FOR PACIFIC WHITE
SHRIMP (Litopenaeus vannamei) REARED IN GREEN WATER SYSTEMS
Elkin A. Amaya
A Thesis
Submitted to
the Graduate Faculty of
Auburn University
in Partial Fulfillment of the
Requirements for the
Degree of
Master of Science
Auburn, Alabama
Aug 7th, 2006
iii
FISH MEAL REPLACEMENT IN PRACTICAL DIETS FOR PACIFIC WHITE
SHRIMP (Litopenaeus vannamei) REARED IN GREEN WATER SYSTEMS
Elkin A. Amaya
Permission is granted to Auburn University to make copies of this thesis at its
discretion, upon request of individuals or institutions and at their
expense. The author reserves all publication rights.
________________________
Signature of Author
________________________
Date of Graduation
iv
VITA
Elkin A. Amaya, son of Nestor Amaya and Martha Rojas, and brother of Edwin,
Lady and Jonathan, was born on February 24th, 1978, in Bogota, Colombia. He was
conferred his Bachelor of Science degree in Animal Science at the ?Universidad Nacional
de Colombia? in Bogota, Colombia in 2002. After working for two years in the areas of
animal production with governmental agencies and animal nutrition with the feed
industry, in 2004 he joined Auburn University to pursue a Master of Science degree in
Aquaculture at the Department of Fisheries and Allied Aquacultures.
v
THESIS ABSTRACT
FISH MEAL REPLACEMENT IN PRACTICAL DIETS FOR PACIFIC WHITE
SHRIMP (Litopenaeus vannamei) REARED IN GREEN WATER SYSTEMS
Elkin A. Amaya
Master of Science, Aug 7th, 2006
(B.S., Universidad Nacional de Colombia, Bogota, Colombia, 2002)
80 Typed Pages
Directed by D. Allen Davis
A series of experimental studies were conducted with Pacific white shrimp,
Litopenaeus vannamei, to evaluate the use of plant protein sources as replacement
ingredients for fish meal in commercially manufactured diets for shrimp reared in green
water environments. In the first study, juvenile (mean weight + S.D., 0.031 + 0.0005g)
shrimp were stocked in 16 0.1-ha low-water-exchange ponds and reared over an 18-week
culture period. Four commercially extruded diets formulated to contain 35% crude
protein and 8% lipids were evaluated. These diets included 16% poultry by-product meal
and varying levels of fish meal (9, 6, 3, and 0%), which was replaced by a combination of
soybean meal and corn gluten meal to replace the protein originating from fish meal. At
vi
the conclusion of the culture period, there were no significant differences (P>0.05) on any
of the production parameters evaluated among the test diets, whereas feeding costs were
significantly (P<0.05) reduced as more plant proteins were included in the diets. Mean
gross production, final weight, FCR and survival were evaluated at the end of the 18-
week culture period, with final values ranging from 5,363 - 6,548 kg/ha, 18.4 - 20.7 g,
1.12 - 1.38 and 84.0 - 94.0 %, respectively. This study demonstrated that fish meal can be
completely substituted by alternative vegetable protein sources in commercially
manufactured shrimp feeds, without negatively compromising the productive and
economic performance of Litopenaeus vannamei reared in ponds. In the second study,
juvenile (mean weight + S.D. 0.74 g ? 0.03, n=30) shrimp were stocked in an outdoor
covered recirculating system composed of 24 800-L tanks with 30 shrimp per tank and
four replicates per treatment. Experimental treatments included four diets with varying
levels of fish meal in the diet (9%, 6%, 3% and 0%), a plant protein-based diet, and a
commercial reference feed. Feeds were commercially extruded and offered as sinking,
extruded pellets designed to contain 35% crude protein and 8% lipids. Production
parameters at the end of the study demonstrated no significant differences (P> 0.05)
among any of the treatments evaluated. Mean net production, final weight, weight gain
(%), FCR and survival were evaluated at the end of the 81-day culture period. Final
values for these parameters ranged from 564.4 - 639.0 g/m3, 17.4 - 19.5 g, 2,249 - 2,465
%, 1.07 - 1.20 and 83.3 - 89.2 %, respectively. These results indicate that fish meal can be
replaced with solvent extracted soybean meal when diets contain 16% poultry by-product
meal. In addition, plant protein sources such as soybean meal, corn gluten meal and corn
vii
fermented solubles can be used without affecting shrimp performance in diets with no
animal protein sources. Feed costs per unit of production indicate that feed costs can be
reduced if fish meal is replaced with alternative protein sources. Findings from these two
studies provide evidence that shrimp reared in green water systems can successfully use
alternative plant based diets with no fish meal. In addition, it indicates that shrimp have
the capacity to effectively use a plant protein based diet without negatively compromising
shrimp performance. The use of alternative shrimp diets is therefore recommended as a
way to reduce pressure and dependance on fish meal and other animal protein sources,
with the subsequent reduction on feed production costs as cheaper, high quality plant
ingredients are included. Provided that these reduced feed costs are effectively transferred
from the feed companies to the shrimp producers, shrimp producers could gain from
better profit margins as cheaper feeds are used. An additional reason for using alternative
plant-based diets is that they open an opportunity for new markets that would be willing
to pay a higher price, for shrimp fed and produced under more ecologically and
sustainable conditions that do not represent a threat to the environment or the human
health.
viii
ACKNOWLEDGMENTS
I would like to thank my advisor Dr. Allen Davis for his support and guidance
throughout my Auburn experience, same as my committee members Dr. William Daniels
and Dr. David Rouse for their guidance. I am especially grateful to my parents Nestor
and Martha and my siblings Edwin Lili and Jonathan for their constant company. I am
also greatly thankful to Evi and her company and support throughout this time in Auburn.
This study was funded by the American Soybean Association (ASA, MASEA
Grant) and the Department of Fisheries and Allied Aquacultures at Auburn University.
Special thanks to all the students of the Nutrition and Technology Laboratory of the
Department of Fisheries and Allied Aquacultures of Auburn University for their help and
support during the development of this study. I also want to thank the Alabama
Department of Conservation and Natural Resources, Marine Resources Division, for
allowing the use of their facilities during the experimental part of this study and for their
physical and logistic support.
ix
Style manual of journal used: Aquaculture
Computer software used: Word Perfect 12, Microsoft Power Point, Microsoft Excel XP,
and SAS v. 9.1
x
TABLE OF CONTENTS
LIST OF TABLES .................................................................................................. xi
LIST OF FIGURES ............................................................................................. xiii
CHAPTERS
I. INTRODUCTION ...................................................................................... 1
II. REPLACEMENT OF FISH MEAL IN PRACTICAL DIETS FOR THE
PACIFIC WHITE SHRIMP (Litopenaeus vannamei) REARED UNDER
POND CONDITIONS.
Abstract.......................................................................................................
Introduction.................................................................................................
Methods.......................................................................................................
Results and Discussion................................................................................
Conclusion...................................................................................................
Aknowledgements.......................................................................................
References...................................................................................................
8
10
12
20
32
33
34
III. ALTERNATIVE DIETS FOR THE PACIFIC WHITE SHRIMP
Litopenaeus vannamei
Abstract.......................................................................................................
Introduction.................................................................................................
Materials and Methods................................................................................
Results.........................................................................................................
Discussion...................................................................................................
Conclusion............................................................................................... ...
Aknowledgements.................................................................................... ..
References...................................................................................................
37
39
41
47
51
54
54
55
IV. SUMMARY AND CONCLUSIONS.......................................................... 59
V. LITERATURE CITED................................................................................ 64
xi
LIST OF TABLES
II. 1 Feeding rates as percentage biomass and feed type utilized through a
19-day nursery period for Litopenaeus vannamei post-larvae. Feed
inputs were based on mean shrimp weight and an assumed 78%
survival.................................................................................................. 14
2 Ingredient composition (g/100 g as is) of practical diets for
Litopenaeus vannamei, used for the replacement of fish meal (FM)
with plant protein sources. Diets were commercially manufactured by
Rangen? Inc.(Angleton, TX) using extrusion processing.................... 17
3 Nutritional composition of practical diets for Litopenaeus vannamei,
used for the replacement of fish meal (FM).......................................... 18
4 Water quality parameters observed over a 19-day nursery period for
Litopenaeus vannamei, stocked at a density of 53 post-larvae/L in a
water recirculating system composed of six, 4000-L nursery tanks...... 21
5 Mean production parameters of Litopenaeus vannamei nursed for 19
days and stocked at a density of 53 post-larvae/L (mean weight +
S.D., 1.48mg + 0.39) in a water recirculating system composed of
six, 4000-L nursery tanks...................................................................... 22
6 Summary of water quality parameters observed over an 18-week
growing period for Litopenaeus vannamei fed four practical diets
with varying levels of fish meal (FM) and cultured in 0.1-ha ponds.
Values are mean ? standard deviation of daily and weekly
determinations. Values in parenthesis represent minimum and
maximum readings of the whole data set.............................................. 24
7 Mean productive parameters of Litopenaeus vannamei cultured in
0.1-ha ponds, at the end of an 18-week culture period and fed four
practical diets with varying levels of fish meal (FM) and plant protein
sources................................................................................................... 26
8 Mean production (kg/ha) of Litopenaeus vannamei fed four practical
diets with varying levels of fish meal (FM) and plant protein sources.
Production is divided in the number of head-on shrimp per lb (454g).
Prices are based on Gulf fresh shrimp prices between the 13 and 19
of October, 2005.................................................................................... 27
xii
9 Mean economic parameters of Litopenaeus vannamei reared in ponds
at the end of an 18-week culture period and fed four practical diets
with varying levels of fish meal (FM) and plant protein sources.
Estimates are based on shrimp production per hectare.......................... 31
III. 1 Ingredient composition of practical diets for Litopenaeus vannamei
used to evaluate the replacement of animal proteins by plant protein
sources (values expressed on an as fed basis, g/100 g). Diets were
commercially manufactured by Ranger? Inc.(Angleton, TX) using
extrusion processing.............................................................................. 44
2 Nutritional composition of practical diets for Litopenaeus vannamei
used to evaluate the replacement of animal proteins with plant
protein sources....................................................................................... 45
3 Summary of water quality parameters observed over an 81-day
experimental period for Litopenaeus vannamei fed practical diets
with varying levels of animal and plant protein sources and cultured
in an outdoor semi-close recirculating culture system. Values are
mean ? standard deviation of daily and weekly determinations.
Values in parenthesis represent minimum and maximum readings
throughout the study.............................................................................. 48
4 Mean production parameters at the end of an 81-day culture period
for Litopenaeus vannamei reared in an outdoor semi-closed
recirculating culture system and fed practical diets with varying
levels of fish meal (FM), a plant based diet and a commercial
reference diet......................................................................................... 49
5 Mean economic parameters of Litopenaeus vannamei cultured in an
outdoor semi-closed recirculating system over an 81-day
experimental period and fed practical diets with varying levels of fish
meal (FM), and a plant protein based diet. Estimates are based on
production per cubic meter of culture system....................................... 50
xiii
LIST OF FIGURES
I. 1 Daily feed inputs (kg/ha/day) for Litopenaeus vannamei raised in
ponds at a density of 35 shrimp/m2 over an 18-week growing period
and fed four practical diets with varying levels of fish meal and plant
protein sources....................................................................................... 25
2 Population percentages of Litopenaeus vannamei, divided on the size
of head-on shrimp count per lb. Shrimp was fed four practical diets
with varying levels of fish meal and raised in ponds for an 18-week
period at a density of 35 shrimp/m2....................................................... 28
1
CHAPTER I
INTRODUCTION
The commercial production of farmed shrimp has been expanding steadily.
According to FAO (2005), 1,804,932 mt (metric tons) of shrimp were cultured in 2003,
representing an increase of around 640,000 mt over the production of 2000. The
contribution of aquaculture to world shrimp production reached 32.5% in the period
between 2001 and 2003 compared to an average of 27.5% during the 1990?s. Most of this
increase was the result of the development of Pacific white shrimp (Litopenaeus
vannamei) aquaculture in Asian countries, such as China and Thailand, and to the
increased production from Brazil in the western hemisphere. Combined, these three
countries produced around 510,000 mt out of the 723,858 mt of the Litopenaeus
vannamei cultured in 2003 (FAO, 2005). This increased production has been
accompanied by a decrease in shrimp value either due to depressed economies and/or
abundant supplies. Expansion of this industry is expected to continue and shrimp market
prices are likely to continue to fall, therefore, reducing the profitability of the industry.
According to Josupeit (2004), shrimp trade has not grown in value in recent years and
since 2000 all unit values of shrimp have gone down with record low prices in 2004.
Facing this scenario, there has been a general interest in finding alternatives to generate
2
an added value to the shrimp industry and to re-evaluate and improve the shrimp
production systems in terms of yields, production costs and feed efficiencies.
When evaluating ways to reduce production costs, one consideration is to
minimize the use of expensive marine animal ingredients in the feeds. Of particular
concern is fish meal, for which high demand and limited supply makes it a costly
ingredient. The interest for exploring alternative protein sources to fish meal in marine
shrimp diets was initially led by the often variable nature of the price, supply and quality
of the fish meal (Colvin and Brand, 1977; Divakaran et al., 2000). Recently, however,
growing concerns about the use of fish to feed fish, and the increasing demand and
limited fish meal supply have heightened this interest. Different authors have also
claimed that a better shrimp value could be achieved in alternative markets, such as
organic markets, which among other characteristics, require limitations of specific
ingredients such as those of animal origin (Davis et al., 2004; Josupeit, 2004).
Fish and marine animal meals are used in aquatic feeds because they are excellent
sources of essential nutrients such as protein and indispensable amino acids. Additionally,
they contain essential fatty acids, cholesterol, vitamins, minerals, attractants and
unidentified growth factors (Swick et al., 1995; Samocha et al., 2004). Because of these
characteristics, fish meal is also the primary and most expensive ingredient in commercial
shrimp feeds, with commercial formulations commonly including between 25% to 50%
of the total diet. (Tacon and Barg, 1998; Dersjant-Li, 2002).
Protein ingredients that can be utilized to partially or completely substitute marine
animal meals include terrestrial animal and plant by-products readily available on world
3
markets. (Samocha et al., 2004). Animal sources are primarily rendered by-products, such
as blood meal, feather meal, meat and bone meal and poultry by-product meal, which
usually contain 45?65% crude protein and are often good sources of indispensable amino
acids. The nutritional composition of these meals, however, is highly variable depending
on both the quality of raw ingredients and the type of processing. Additionally, due to
events of BSE (mad cow disease) and contaminants such as PCBs, there is an increasing
public concern in the use of potentially contaminated by-products in animal feeds
(Dersjant-Li, 2002; Samocha et al., 2004).
Because of their low price, relatively consistent nutrient composition and supply,
plant proteins, such as oil seed cakes, are often economically and nutritionally valuable
sources of protein. However, due to potential problems associated with insufficient levels
of indispensable amino acids (e.g., lysine and methionine), anti-nutritional factors and
poor palatability, their commercial use has been often limited. Among plant protein
sources, soybean meal has received considerable attention as a replacement for fish meal
in aquatic feeds because of its balanced amino acid profile, consistent composition,
ample availability and price (Lim et al., 1998; Hardy, 1999; Samocha et al., 2004).
Divakaran et al. (2000) showed that the apparent digestibility of protein in soybean meal
is at least 89% in Litopenaeus vannamei.
Common concerns regarding the use of soybean meal are related to the presence
of anti-nutritional factors, such as trypsin inhibitors, which interfere with protein and lipid
digestion. Other anti-nutritional factors, such as antigens, lectins, saponins, and
oligosaccharides negatively affect the palatability and nutrient absorption of feeds
4
containing soybean meal (Dersjant-Li, 2002). According to Swick et al. (1995) the dietary
limitation on soybean meal for shrimp is mostly due to negative effects on the physical
stability of the pellets caused by fiber and soluble sugars. Additionally, most of the
phosphorus in soybean meal is bound to phytic acid and only 30-40% of the total
phosphorus content is considered to be available for Litopenaeus vannamei. (Akiyama et
al. 1991; Hertrampf and Piedad-Pascual, 2000). According to Akiyama (1988),
phosphorous is the most critical mineral when formulating fish feeds that contain a high
level of soy protein. When compared to fish meal, soybean meal is also characterized by
its lower composition of essential amino acids mainly methionine, lysine and threonine
(NRC, 1993), and by its lack of n-3 marine fatty acids (eicopentaenoic acid, EPA and
docosohexaenoic acid, DHA), which are essential for the growth and survival of marine
shrimp (Fox et al., 2004)
Studies evaluating soybean meal as the only replacement of marine animal
proteins in diets for Litopenaeus vannamei have shown a moderate level of success. Lim
and Dominy (1990) reported that up to 40% of a marine protein mix (53% menhaden fish
meal, 34% shrimp waste meal and 13% squid meal) could be replaced by solvent-
extracted soybean meal, without reducing the growth of Litopenaeus vannamei. At this
substitution level, soybean meal represented 28% of the diet, similar to the 10% to 20%
that Akiyama et al. (1991) reported as the percentage typically included in commercial
shrimp feeds. A 25-30% dietary level of soybean meal as a replacement for 40-50% of the
marine animal protein in shrimp feeds appears optimal (Swick et al., 1995). According to
Samocha et al. (2004), a promising alternative to using terrestrial plant or animal proteins
5
independently would be to use a mixture of complementary ingredients to increase
nutrient utilization and facilitate feed processing. Under this strategy, it is possible to
obtain a more balanced nutrient profile in the feeds (i.e. essential amino acids, fatty acids)
than when limited ingredients are used.
Recent studies of Davis and Arnold (2000) and Samocha et al. (2004) have
provided evidence that co-extruded soybean and poultry by-product meal are adequate
ingredients to substitute fish meal in practical diets for Litopenaeus vannamei. Davis and
Arnold (2000) reported that up to 80% of the fish meal in diets for Litopenaeus vannamei
can be substituted either by co-extruded soybean poultry by-product meal containing egg
supplement or poultry by product meal without any apparent effect on shrimp survival or
growth. Samocha et al. (2004) did not find significant differences in the weight gain,
survival and feed efficiency values (FE) of Litopenaeus vannamei juveniles fed practical
diets with 32% protein, where up to 100% of the fish meal was replaced with co-extruded
soybean poultry by-product meal with egg supplement. The authors suggested that the
favorable response of the shrimp to the diets was probably due to the high quality of the
ingredients in terms of nutrient profile, digestibility and lack of palatability problems.
They also reported that there were no indications of the feed being rejected. Furthermore,
they recognized that co-extrusion was a valuable practice when processing the
ingredients.
Based on these findings, it is clear that by using the single ingredient approach,
the complete fish meal replacement with soybean meal is difficult to achieve. Yet, it also
appears that fish meal and marine oil sources can be removed from shrimp diets if
6
suitable alternative sources of protein and lipids are provided to meet the essential amino
acid and fatty acid requirements of shrimp (Davis et al., 2004). Under this scenario and
considering recent advancements in additives and ingredients and feed processing
technologies, it appears that soybean meal use can be further increased in shrimp feeds.
According to Dersjant-Li (2002), properly processed plant ingredients containing high
protein content with high digestibility of crude protein and low anti-nutritional
components are potential protein sources for the replacement of fish meal in fish and
shrimp diets. Today, for example, extrusion is widely used in manufacturing shrimp
feeds, having the advantage of inactivating and/or destroying endogenous heat-sensitive
anti-nutritional factors commonly found in soybean meal and gelatinizing starch granules
(Carver et al., 1989). Amino acid micro-encapsulation is another technology that may
increase the use of synthetic amino acids to correct nutritional imbalances in shrimp
feeds. According to Swick et al. (1995) micro-encapsulation prevents the rapid elevation
of amino acid concentration in shrimp tissue and reduces the solubility problems of non-
encapsulated sources.
Nutritional studies evaluating soybean meal have often been conducted in
controlled, laboratory conditions to mitigate the effects of variable water quality
variations found in shrimp ponds and remove external food sources (D'Abramo and
Castell, 1997). The practical application of data from these studies is limited because
laboratory conditions are different from the shrimp pond environment (Tacon, 1996).
Growth enhancement of shrimp in ponds has commonly been attributed to their
assimilation of micro-algae and microbial-detrital aggregates present in the water (Moss
7
and Pruder, 1995). Furthermore, according to Moss et al. (2001), the culture environment
may also significantly impact the digestive enzyme activity in shrimp, providing the
shrimp with additional digestive capabilities, which may contribute to the growth-
enhancing effect of shrimp in green water systems.
With increasing interest on the applicability of research results under commercial
shrimp production conditions, the primary objective of this study was to evaluate the
replacement of fish meal by using a combination of plant and animal protein sources in
commercially-manufactured diets for L vannamei reared under production conditions in
green water systems. To achieve this objective, two separate studies were carried out. In
the first study, four diets with a fixed level of poultry by-product meal and varying levels
of fish meal and plant protein sources were evaluated in shrimp reared in a pond
production system with minimal water exchange. In the second study, shrimp reared in a
green water recirculating system were used to evaluate the previous four diets in addition
to a plant-based diet with no animal protein sources and a commercial reference diet. It is
expected that because of the commercial and practical scale of this research, findings
from this study will further increase the knowledge about the nutrition and feeding of
shrimp fed diets with high levels of plant protein sources, and will provide the feed and
shrimp industries with additional understanding of marine shrimp capacity to use these
kind of ingredients in the feeds.
8
CHAPTER II
REPLACEMENT OF FISH MEAL IN PRACTICAL DIETS FOR THE PACIFIC
WHITE SHRIMP (Litopenaeus vannamei) REARED UNDER POND CONDITIONS.
Abstract
Increasing economical and ecological concerns regarding the use of fish meal in
diets for marine shrimp have led to the development of replacement strategies where
soybean meal has received ample attention. Most studies evaluating these strategies have
been carried out under laboratory conditions which greatly differ from production
conditions in ponds. This study evaluated a fish meal replacement strategy using
vegetable protein sources in practical feeds for marine shrimp reared in ponds. Juvenile
Pacific white shrimp (Litopenaeus vannamei) (mean weight + S.D., 0.031 + 0.0005g)
were stocked into 16 0.1-ha low-water-exchange ponds and reared over an 18-week
period. Four commercially extruded diets formulated to contain 35% crude protein and
8% lipids were evaluated. These diets included varying levels of fish meal (9, 6, 3, and
0%) which was replaced by a combination of soybean meal (32.48, 34.82, 37.17 and
39.52% respectively) and corn gluten meal (0.00, 1.67, 3.17, and 4.84% respectively) to
replace the protein originating from fish meal. At the conclusion of the experimental
period, there were no significant differences (P>0.05) in shrimp production among the
9
test diets. Mean final production, final weight, FCR and survival were evaluated at the
end of the 18-week culture period, with final values ranging from 5,363 - 6,548 kg/ha,
18.4 - 20.7 g, 1.38 - 1.12 and 84.0 - 94.0 %, respectively. Although not significant, the
economic analysis numerically showed a general increase in the partial gross returns of
shrimp production, as higher levels of plant proteins sources were included in diets fed to
marine shrimp. Results from this study demonstrate that fish meal can be replaced using
alternative vegetable protein sources in practical shrimp feeds without compromising
productive and economic performance of Litopenaeus vannamei reared in ponds.
Keywords: Litopenaeus vannamei, Soybean meal, Fish meal, Plant proteins.
10
Introduction
The commercial production of farmed shrimp has been expanding steadily.
According to FAO (2005), 1,804,932 mt (metric tons) of shrimp were cultured in 2003,
representing an increase of around 640,000 mt over the production of 2000. This
increased production has been accompanied by a decrease in shrimp price, either by
depressed markets or overproduction. With shrimp aquaculture expected to continue to
increase in coming years, shrimp prices are likely to continue to fall as production
exceeds demand, therefore challenging the profitability of this industry.
An important issue considered to reduce shrimp production costs and increase
producers profitability, is the use of feeds with low levels of fish meal and high levels of
cheaper, high quality plant protein sources. Commercial shrimp formulations commonly
include between 25% to 50% of fish meal, representing the primary and most expensive
protein ingredient. (Dersjant-Li, 2002; Tacon and Barg, 1998). Fish meal is preferred
among other protein sources because it is an excellent source of essential nutrients such
as protein and indispensable amino, essential fatty acids, cholesterol, vitamins, minerals,
attractants and unidentified growth factors (Swick et al., 1995; Samocha et al., 2004).
However, limited availability and high demand make of fish meal a high cost ingredient.
Because of their low price and consistent nutrient composition and supply, plant
proteins, such as oil seeds, are often economically and nutritionally valuable alternatives
to fish meal. Among plant protein sources, soybean meal has received considerable
attention in the replacement of fish meal in aquatic animal feeds because of its balanced
amino acid profile, consistent composition, worldwide availability and lower price
11
(Colvin and Brand, 1977; Lim and Dominy,1990; Akiyama et al. (1991; Swick et al.,
1995; Lim et al., 1998; Hardy, 1999; Divakaran et al., 2000; Samocha et al., 2004).
However, when compared to fish meal, soybean meal is characterized by a lower
composition of essential amino acids, mainly methionine, lysine and threonine (NRC,
1993), and by its lack of the n-3 marine fatty acids EPA (eicopentaenoic acid) and DHA
(docosohexaenoic acid), which are essential for the growth and survival of marine shrimp
(Fox et al., 2004). Additionally, only 30-40% of the total phosphorus content is
considered to be available for Litopenaeus vannamei (Hertrampf and Piedad-Pascual,
2000; Akiyama et al., 1991). If the replacement strategy considers shifts in essential
nutrients, it also appears that fish meal can be removed from shrimp formulations if
suitable alternative sources of protein and lipids are provided to meet the nutritional
requirements of the animal (Davis et al., 2004). The use of complementary ingredients is
a practice used to obtain a more balanced nutrient profile in the feeds (i.e. essential amino
acids, fatty acids) and to increase nutrient utilization and facilitate feed processing
(Mendoza et al., 2001; Hernandez et al., 2004; Samocha et al. 2004). Davis and Arnold
(2000), reported that up to 80% of the fish meal in diets for Litopenaeus vannamei can be
substituted by co-extruded soybean poultry by-product meal containing egg supplement
or poultry by-product meal without any apparent effect on survival, growth and feed
palatability. Samocha et al. (2004) did not find significant differences in weight gain,
survival and feed efficiency of Litopenaeus vannamei fed 32% CP (crude protein)
practical diets, where up to 100% of fish meal was replaced with co-extruded soybean
poultry by-product meal with egg supplement.
12
Studies evaluating soybean meal and other alternative protein sources to fish meal
have often been conducted in controlled laboratory conditions to mitigate the inherent
effects of water quality and natural food sources found in shrimp ponds. Although these
studies have provided valuable information regarding shrimp capacity to utilize these type
of ingredients under controlled conditions, the practical application of data from these
studies is limited. Main concerns are related to differences in the culture environment
(e.g. green water ponds vs clear water tanks), length of the growth cycle, and feed
processing (extruded vs pelleted) (Moss and Pruder, 1995; Tacon, 1996; D'Abramo and
Castell, 1997; Moss et al., 2001). As a normal transition to an effective process of
technology transfer aimed to reach the shrimp and feed industries, the next step would be
to carry out studies under conditions similar to those found commercially. This study
evaluates fish meal replacement using soybean meal as the main protein source in
commercially manufactured feeds for the Pacific white shrimp Litopenaeus vannamei
reared under pond conditions.
Methods
Shrimp source
Pacific white shrimp Litopenaeus vannamei post-larvae (mean weight + S.D.,
1.48mg + 0.39) were obtained from Shrimp Improvement Systems (Plantation Key, FL)
and nursed for 19 days in a recirculation system at the Claude Peteet Mariculture Center
located in Gulf Shores, Alabama, according to the procedures described by Garza et al.
(2004). The nursery system was stocked at a density of 53 post-larvae per liter and the
13
following water quality parameters maintained at adequate levels for the growth of
shrimp post-larvae: temperature, dissolved oxygen, pH, salinity and total ammonia
nitrogen. Feeding schedule for shrimp during the nursery period is presented in Table 1.
At the conclusion of the nursery phase, juvenile shrimp (0.0312g + 0.0005) were stocked
into 16 ponds located in the same facilities, at a density of 35 shrimp per squared meter.
Pond management and water quality
Ponds used for the grow-out phase were 0.1 ha in surface area (46 x 20 m), 1.0 m
average depth, and lined with a 1.52 mm thick, high-density polyethylene. Each pond
bottom was covered with a 25-cm deep layer of sandy-loam soil and equipped with a 20-
cm diameter screened standpipe and a concrete catch basin. Prior to use, pond soils were
dried and tilled to allow oxidation and mineralization of organic matter. Two weeks
before stocking, ponds were filled with brackish water (9-13 ppt) from the Intracostal
Canal between Mobile and Perdido Bay, Alabama. Inlet water was filtered through a 250-
?m nylon filter sock (Micron Domestic Lace Mfg., Inc) in order to prevent the
introduction of predators and minimize the introduction of larval species. Inorganic liquid
fertilizers were mixed and applied one week before stocking at a rate of 1,697 ml of 32-0-
0 and 303 ml of 10-34-0 per pond, therefore, providing 5.73 kg of N and 1.03 kg of
P205/ha. Two weeks after the first fertilization, a second fertilizer application at half the
initial rate was added to ponds with Sechii disk readings greater than 50 cm. Twenty-four
hours before stocking, a 1:15 mixture of motor oil and diesel fuel was applied to the pond
surface, at a rate of 900 ml per pond, to reduce the number of air breathing insects.
14
Table 1. Feeding rates as percentage biomass and feed type utilized through a 19-day
nursery period for Litopenaeus vannamei post-larvae. Feed inputs were based on mean
shrimp weight and an assumed 78% survival.
Day Shrimp weight
(mg)
Feed rate
(%)
Feed type Ratio
(%)
1-3 1.5 25 Artemia / PL redia 40 / 60
4-6 2.5 25 PL redi / Crumble #0b 40 / 60
7 4.1 25 PL redi / Crumble #0 20 / 80
8-9 4.1 25 Artemia / Crumble #0 5 / 95
10-12 5.7 15 Artemia / Crumble #0 15 / 85
13-15 11.9 15 Crumble #0 / Crumble #1b 50 / 50
16 20.1 15 Crumble #0 / Crumble #1 30 / 70
17-18 20.1 15 Crumble #1 100 / 0
19 28.9 15 Crumble #1 /Crumble #3b 50 / 50
Harvest 31.2
a PL Redi-reserve 400-600 microns. 50% Protein, Zeigler Bross, Inc., Gardners, PA, USA
b Rangen 45% protein, Rangen Inc., Buhl, Idaho, USA.
15
During the experimental period, dissolved oxygen, temperature, salinity and pH
concentrations were measured at sunrise (05:00-05:30) and at night (20:00-22:00) using a
YSI 556MPS meter (Yellow Spring Instrument Co., Yellow Springs, OH, USA). Total
ammonium-nitrogen (TAN) and Sechii disk readings were determined on a weekly basis.
Water samples for TAN analysis were taken at 40 cm from the pond surface and measured
with a spectrophotometer (Spectronic Instrument Inc. Rochester, NY, USA) by the
Nesslerization method (APHA 1989).
In order to maintain minimum dissolved oxygen levels of 3 mg/L, each pond was
provided with a base aeration capacity of 30 hp/ha (7.5kW/ha). Paddle wheel aerators of 1-
hp (Little John Aerator, Southern Machine Welding Inc. Quinton, AL) or propeller
aspirators aerators of 1-hp (11.2 Ampers) or 2-hp (20 Ampers) (Aire-O2, Aeration
Industries International, Inc. Minneapolis, Minnesota) were used for this purpose. When
required, additional aeration (up to 30 hp/ha) was used to maintain adequate DO levels.
Dissolved oxygen and temperature stratification within the water column after the 8th week
were managed by running the aerators for about 20 minutes in the late afternoon. Ponds
were managed with a minimal water exchange strategy; therefore, there were no regular
water exchanges. However, because of heavy rains associated with tropical storms, the
water level of the pond had to be reduced by 20%, twice at different times of the crop to
avoid the risk of flooding. Additionally, the week prior to harvest, 30% of the water from
the ponds was replaced, in order to reduce the chance of algae crashes and to encourage
shrimp molting before harvest (Davis, personal communication).
16
Feed formulation strategy and feed management
Shrimp ponds were randomly assigned to one of four dietary treatments, with four
replicates per treatment. The basic guideline for the formulation of the four diets was to
reduce the inclusion of menhaden fish meal (FM) as follows: 9%FM, 6%FM, 3%FM and
0%FM of the total diet, while increasing the inclusion of vegetable protein sources, mainly
solvent extracted soybean meal (Table 2). The dietary treatments were formulated in order
to provide equal protein and lipid levels while maintaining a minimum lysine and
methionine plus cystine content of 5% and 3% of the total protein, respectively (Table 3).
Corn gluten meal was used as a natural source of methionine. Lipid levels were adjusted
by adding menhaden fish oil. Calcium phosphate was added to ensure adequate
phosphorus supply as fish meal was removed. Feeds were produced by Rangen, Inc.
(Angleton TX, USA) under commercial manufacturing conditions and offered twice a day
to the shrimp, as a sinkable, extruded 3 mm pellet.
Feed inputs during the 1st, 2nd, 3rd and 4th weeks of pond culture were set at 10, 15,
30 and 60 kg of feed/ha/day, respectively. For the remainder of the growth trial, feed
inputs were back-calculated, based on an expected weight gain of 1.5 g per week, a feed
conversion of 1.2:1, and a total mortality of 30% (1.66%/wk) over an 18-weeks culture
period. Feeding during the first 19 days after stocking was done using the same
commercial 35% CP Rangen feed that had been used at the end of the nursery phase.
17
Table 2. Ingredient composition (g/100 g as is) of practical diets for Litopenaeus vannamei
used for the replacement of fish meal (FM) with plant protein sources. Diets were
commercially manufactured by Rangen? Inc.(Angleton, TX) using extrusion processing.
Ingredient 9% FM 6% FM 3% FM 0% FM
Soybean meal 32.48 34.82 37.17 39.52
Fish meal - Menhaden 9.00 6.00 3.00 -
Poultry By-Product meal 16.00 16.00 16.00 16.00
Milo 35.47 33.82 32.33 30.68
Corn Gluten meal - 1.67 3.17 4.84
Fish Oil 3.96 4.22 4.47 4.72
Di-calcium Phosphate 1.50 1.88 2.27 2.65
Bentonite 1.00 1.00 1.00 1.00
Mold Inhibitor 0.15 0.15 0.15 0.15
Vitamin Premix 0.34 0.34 0.34 0.34
Mineral Premix 0.08 0.08 0.08 0.08
Stay-C 35% 0.02 0.02 0.02 0.02
18
Table 3. Nutritional composition of practical diets for Litopenaeus vannamei used for the
replacement of fish meal (FM)1.
Ingredient 9% FM 6% FM 3% FM 0% FM
Crude Protein 35.7 35.9 36.2 36.6
Crude Fat 8.4 8.3 8.6 8.4
Crude Fiber 2.4 1.8 2.1 1.9
Ash 8.2 7.9 7.9 8.1
Calcium2 1.3 1.3 1.2 1.1
Total Phosphorus2 1.2 1.2 1.2 1.3
Lysine2 2.0 2.0 1.9 1.8
Met + Cys2 1.1 1.1 1.1 1.1
% Protein from plant sources2,3 54.9 60.4 66.0 71.5
% Protein from animal sources2,4 45.1 39.6 34.0 28.5
1 Analyzed value - New Jersey Feed Laboratory, Inc. Trenton, NJ.
2 Calculated value.
3 Soybean meal, corn gluten meal, milo.
4 Fish meal, poultry by-product meal.
19
Treatment feeds were offered from day 20, when it was estimated that shrimp had
reached 1 g of weight. Maximum feed inputs were set at 83.3 kg of feed/ha/day at the 5th
week. Feed inputs throughout the study are shown in Figure 1. Shrimp growth was
monitored on a weekly basis by determining the average weight in a sample of 70 to 100
animals per pond. Sampling was carried out by capturing shrimp by seine during first two
weeks and cast net (monofilament net, 1.22 m radius and 0.95 cm opening) for the
remaining of the culture period.
Harvest
Harvest took place over a three day period, after 18 weeks of pond culture. Feed
inputs were stopped two days prior to harvesting a given pond. The night before harvest,
two thirds of the water from each pond was drained and aeration was provided using
paddlewheel aerators to keep shrimp alive and minimize erosion on pond bottoms. On
harvest day, shrimp were pumped from the catch basin using a hydraulic fish pump
equipped with a 25-cm diameter suction pipe (Aqualife-Life pump, Magic Valley Heli-arc
and Mfg, Twin Falls, Idaho, USA). The pump was placed in the catch basin and shrimp
were pumped, de-watered and collected in a hauling truck. Shrimp were then transferred to
a laboratory located at the same facilities for washing, cleaning and weighing. During
weighing a random sample of 125 shrimp was collected for individual weight
determinations. Individual weights were used to calculate mean final production, mean
final weight, survival and size distributions. After quantifying the biomass from each
pond, mean final production (final biomass), feed conversion ration (total feed offered /
20
biomass increase), size distribution and survival were determined. Additional economic
considerations about production and feeding costs were also calculated.
Data Analysis
Data were analyzed using an analysis of variance to determine significant (p<0.05)
differences among treatment means. The Student?Neuman?Keuls multiple comparison
test was used to determine significant differences between treatment means (Steel and
Torrie, 1980). All statistical analyses were conducted using SAS (V9.1., SAS Institute,
Cary, NC, USA).
Results and Discussion
Water quality parameters throughout the 19 days nursery phase maintained suitable
levels for the adequate growth and survival of juvenile shrimp (Tables 4 and 5). Prior to
stocking of the nursed shrimp in the ponds, Sechii disk readings were measured, with
values ranging from 70-110 cm. These values were considerably low as compared to
readings at stocking from previous research at the same facilities (Venero, 2006; Zelaya,
2005) where they obtained readings between 25-85 cm. Low readings at stocking were the
result of only having one week from fertilization to stocking, as compared to four weeks in
the other studies. By the 5th week after stocking, it was clear that plankton blooms had
completely developed with Sechii disk readings ranging from 25-35 cm. Weekly
fluctuations in Sechii disk readings were the results of plankton bloom and die-off cycles in
the ponds.
21
Table 4. Water quality parameters observed over a 19-day nursery period for Litopenaeus
vannamei stocked at a density of 53 post-larvae/L in a water recirculating system composed
of six, 4000-L nursery tanks.
Parameter Mean Minimum Maximum Standard
Deviation
CVa
Temperature (?C) 28.0 24.3 30.3 1.38 4.93
Dissolved Oxygen (mg/L) 6.18 4.76 7.82 0.45 7.30
pH 7.50 6.97 7.83 0.21 2.80
Salinity (ppt) 28.0 13.8 31 4.54 16.19
TAN b (mg/L) 0.63 0.00 1.78 0.56 89.50
a CV = Standard deviation / mean * 100.
b Total Ammonium-Nitrogen.
22
Table 5. Mean production parameters of Litopenaeus vannamei nursed for 19 days and
stocked at a density of 53 post-larvae/L (mean weight + S.D., 1.48 + 0.39mg) in a water
recirculating system composed of six, 4000-L nursery tanks.
Parameter Mean Minimum Maximum Standard
Deviation
CVa
Final weight (mg/shrimp) 31.2 27.2 37.2 3.6 11.61
Weight Gain (%) 1,950 1,740 2,415 249 12.78
Survival (%) 74.2 61.5 79.6 7.1 9.56
FCRb 1.04 1.00 1.09 0.03 2.85
Final biomass (kg/m3) 1.18 1.13 1.22 0.03 2.41
a Coefficient of variation = Standard deviation / mean * 100.
b Apparent feed conversion ratio = Total weight of feed given / Biomass increase.
23
Water quality parameters throughout the 18-week experimental period maintained
suitable levels for adequate growth and survival of shrimp (Table 6). Analysis of mean
values for water quality parameters showed no statistically significant differences among
treatments. Specific events of quick reduction in DO concentrations were observed during
the second half of the study. These were principally associated to algae die-offs, which
subsequently increased TAN concentrations and oxygen demand. Whenever an algae die-
off was noticed, two aerators were set to operate continuously for as long as it was required
(typically 3-4 days), until a new bloom could develop.
The mean number of dissolved oxygen readings that fell below a critical value of
2.5 mg/L, which is considered to initiate stressful conditions in shrimp (Venero 2006;
Zelaya 2005), are presented in Table 6. Potentially stressful pH values below 6 units were
not identified, whereas pH readings greater than 10 units were identified only four times.
Analysis of extreme dissolved oxygen and pH values demonstrated no statistically
significant differences among any of the treatments evaluated.
Feed inputs were modified from the original feeding program when conditions such
as low dissolved oxygen levels, risk of storm or lack of electric power to run the aerators
were present. Final daily feed inputs throughout the study are shown in Figure 1. Mean
productive parameters of Litopenaeus vannamei fed the four dietary treatments at the end of
the experimental period are presented in Table 7. Mean final productions (kg/ha) divided
into production per count size of head-on shrimp are presented in Table 8. Distribution
percentages of shrimp divided by count size of head-on shrimp for all treatments are
presented in Figure 2.
24
Table 6. Summary of water quality parameters observed over an 18-week growing period
for Litopenaeus vannamei fed four practical diets with varying levels of fish meal (FM) and
cultured in 0.1-ha ponds. Values are mean ? standard deviation of daily and weekly
determinations. Values in parenthesis represent minimum and maximum readings of the
whole data set.1
Parameter 9% FM 6% FM 3% FM 0% FM
DO (mg/l)2
am 4.12 ? 1.64
(0.06, 13.28)
4.09 ? 1.59
(0.13, 11.56)
4.22 ? 1.48
(0.59, 12.83)
4.01 ? 1.58
(0.78, 12.62)
pm 7.25 ? 2.43
(0.97, 16.57)
6.92 ? 2.48
(0.08, 16.80)
7.11 ? 2.42
(0.66, 17.15)
7.31 ? 2.39
(0.82, 16.98)
Readings < 2.5 16.00 ? 7.87 20.00 ? 10.64 11.00 ? 1.82 16.75 ? 6.29
Temperature (?C)
am 29.1 ? 1.6
(24.5, 32.9)
29.2 ? 1.6
(24.5, 33.0)
29.1 ? 1.7
(24.5, 32.7)
29.2 ? 1.6
(24.4, 32.7)
pm 30.6 ? 1.6
(25.3, 34.0)
30.7 ? 1.6
(25.1, 34.4)
30.6 ? 1.6
(25.2, 34.0)
30.8 ? 1.7
(24.7, 34.8)
pH
am 7.61 ? 0.87
(6.16, 9.69)
7.40 ? 0.83
(6.17, 9.62)
7.60 ? 0.87
(6.10, 9.47)
7.41 ? 0.85
(6.12, 10.27)
pm 8.15 ? 0.74
(6.54, 10.04)
8.25 ? 0.77
(6.50, 10.73)
8.41 ? 0.72
(6.87, 9.84)
8.30 ? 0.73
(6.85, 10.02)
Readings > 10 0.25 ? 0.50 0.25 ? 0.50 - 0.50 ? 0.57
Salinity (g/l)
9.79 ? 1.2
(7.4, 13.3)
9.62 ? 1.2
(7.4, 13.8)
9.57 ? 1.2
(7.3, 12.9)
8.93 ? 1.3
(5.6, 11.9)
Turbidity (cm)
43.7 ? 32.6
(14, 110)
46.1 ? 34.2
(10, 110)
43.6 ? 33.2
(14, 110)
44.5 ?35.5
(10, 110)
TAN (mg/l )3
0.77 ? 1.50
(0, 9.33)
1.05 ? 1.68
(0, 7.65)
0.98 ? 1.74
(0, 9.01)
1.15 ?1.86
(0, 9.88)
1 Based on analysis of variance (ANOVA) no significant differences (P > 0.05) were found
among treatment means (n = 4).
2 Dissolved Oxygen.
3 Total Ammonium Nitrogen. Measured once per week.
25
Figure 1. Daily feed inputs (kg/ha/day) for Litopenaeus vannamei raised in ponds at a
density of 35 shrimp/m2 over an 18-week growing period and fed four practical diets with
varying levels of fish meal and plant protein sources.
0
10
20
30
40
50
60
70
80
90
100
0 7 14 21 28 35 42 49 56 63 70 77 84 91 98 105 112 119
Day
Fe
ed
(k
g/h
a./
da
y)


.





26
Table 7. Mean productive parameters of Litopenaeus vannamei cultured in 0.1-ha ponds, at
the end of an 18-week culture period and fed four practical diets with varying levels of fish
meal (FM) and plant protein sources1.
Parameter 9% FM 6% FM 3% FM 0% FM PSE2 P Value
Final weight (g) 19.6 18.4 19.8 20.7 1.39 0.695
Weight gain (%)3 62,603 58,635 62,708 67,219 4,691 0.652
Production (Kg/pond) 584.7 536.3 654.8 634.7 36.1 0.145
Weight gain (g/week) 1.11 1.04 1.13 1.19 0.079 0.695
FCR4 1.24 1.38 1.12 1.14 0.078 0.130
Survival (%) 87.2 84.0 94.0 87.4 5.72 0.661
1 Based on analysis of variance (ANOVA) no significant differences (P > 0.05) were found
among treatment means (n = 4).
2 Pooled standard error of treatment means = mse n
3 Weight gain (%) = 100 x (final weight - initial weight) / initial weight.
4 Apparent feed conversion ratio = Total feed offered / biomass increase.
27
Table 8. Mean production (kg/ha) of Litopenaeus vannamei fed four practical diets with
varying levels of fish meal (FM) and plant protein sources. Production is divided in the
number of head-on shrimp per lb (454g). Prices are based on Gulf fresh shrimp prices
between the 13 and 19 of October, 20051.
Count of head-
on shrimp/Lb
9% FM 6% FM 3% FM 0% FM USD/Lb
< 15 12 - 13 13 2.00
15-20 1,169 311 1,375 1,726 1.80
21-25 1,555 2,360 2,423 3,097 1.65
26-30 2,070 1,781 1,912 1,244 1.60
31-35 854 697 563 254 1.55
36-40 164 118 157 - 1.50
41-50 12 96 92 13 1.40
51-60 11 - - - 1.20
61-70 - - 13 - 1.10
> 70 - - - - 0.95
Total Kg/ha 5,847 5,363 6,548 6,347
1 Reported by the National Marine Fisheries Service, Fisheries Statistics Division, Silver
Spring, MD, USA.
28
Figure 2. Population percentages of Litopenaeus vannamei, divided on the size distribution
of head on shrimp count per lb. Shrimp was fed four practical diets with varying levels of
fish meal and raised in ponds for an 18-week period at a density of 35 shrimp/m2.
0
5
10
15
20
25
30
35
40
45
50
%
<15 16-20 21-25 26-30 31-35 36-40 41-50 51-60 61-70 >70
Shrimp count per lb
9% FM 6% FM
3% FM 0% FM
29
No significant differences were found for any of the production parameters
evaluated. Final weights ranged from 18.4 - 20.7 g and were numerically higher in
treatment 4 (0% FM); whereas mean final production, FCR and survival were higher in
treatment 3 (3% FM) with values ranging for all treatments between 5,363 - 6,548 kg/ha,
1.12 - 1.38 and 84% - 94%, respectively. High mean survival rates suggest appropriate use
of the available food and healthy water quality. These results also indicate that feed inputs
were adequately assimilated, allowing the shrimp to achieve consistent productions and low
feed conversion ratios. Production results obtained in this study are within the production
range of commercial shrimp production in semi-intensive production systems. At the same
facilities and with similar pond management, these production results were similar to those
reported by Venero (2006) and better than those obtained by Zelaya (2005) and Garza
(2004). Commercial shrimp feeds are commonly reported to include fish meal levels
between 25% and 50% of the total diet (Dersjant-Li, 2002; Tacon and Barg, 1998).
However, recent studies have shown that commercial shrimp feeds containing 30-35%
crude protein can include levels as low as 7.5-12.5% fish meal without compromising
shrimp performance (Fox et al., 2004). The inclusion of soybean and corn gluten meals for
the partial and total replacement of fish meal protein in the present study resulted in no
adverse affects on the performance of shrimp. Considering that all the test diets had a fixed
level of poultry by-product meal (16%), it can be concluded that up to 71.5 % of the dietary
protein can be provided by high quality plant protein sources (0% FM feed) in shrimp diets
(Table 3). One of the options proposed to significantly reduce the production costs of the
shrimp industry is to replace fish meal with vegetable protein sources in the formulation,
30
therefore reducing the cost of the feeds. In this study, feed costs were slightly reduced as
more plant proteins were included in the formulation. When evaluated in terms of the total
cost of the feed inputs per treatment, significant differences were found among the
treatments, indicating that under the conditions of this study, feeding cost were effectively
reduced by increasing the level of plant proteins in the diet (Table 9). On the other hand,
there were no significant differences in the partial gross returns obtained when subtracting
the total costs of the feed inputs from the total value of the final production. Thus, at
current ingredient prices and at the substitution levels evaluated in this study, plant protein
substituted for fish meal in a properly formulated and manufactured feed is a cost effective
practice. If production costs are to be reduced, shrimp producers must not only have well
balanced diets but they must also feed them properly. This means that overfeeding must be
minimized and avoid nutrient loading that goes beyond the assimilation capacity of the
culture system. In addition, in cases where the animal has the potential of using natural
productivity, the producer must take advantage of this capacity as a way to improve FCR. It
is generally accepted then, that besides feed inputs, growth and feed utilization by shrimp
reared in minimal water exchange pond systems are influenced by the shrimp ability to
consume the microbial organisms produced within the culture system (Tacon et al., 2002).
Thus, it is clear that the nutrition and feeding of shrimp must be studied under conditions
which mimic as closely as possible those of the intended farm production unit and
environment (Tacon 1996). Likewise, technologies aimed to reach feed and shrimp
producers require the delivery of information produced under conditions comparable to
those used commercially.
31
Table 9. Mean economic parameters of Litopenaeus vannamei reared in ponds at the end of
an 18-week culture period and fed four practical diets with varying levels of fish meal (FM)
and plant protein sources1. Estimates are based on shrimp production per hectare1.
Parameter 9% FM 6% FM 3% FM 0% FM PSE P Value
Final production (Kg/ha) 5,847 5,363 6,548 6,347 360.8 0.145
Production value (USD/ha)2 21,134a 19,131a 23,889a 23,445a 1,562 0.169
Total feed inputs (kg/ha) 7,244a 7,232a 7,244a 7,232a 13.5 0.835
Feed price (USD/kg)3 0.531 0.526 0.52 0.515 - -
Total cost of feed inputs (USD)4 3,849a 3,802b 3,769c 3,723d 7.2 <0.0001
Partial gross returns (USD)5 17,285a 15,328a 20,119a 19,722a 1559 0.159
1 Means (n = 4) not sharing a common supercript within a row are significantly different
(P< 0.05) based on Student Newman-Keuls multiple range test.
2 Calculated based on prices and size distribution from Table 8.
3 Price on May 2005. Subject to change based on varying ingredients price. Prices were
based on those at the time of the research. As fish meal has gone up in price and prices vary
from region to region the cost effectiveness of the feed will change. Hence, conclusion with
regards to cost effectiveness should be made on a case by case basis.
4 Total feed inputs cost = Total Feed Inputs (kg/ha) * Feed price (USD/kg).
5 Partial Gross Returns = Production value (USD) - Total Feed Cost (USD/kg).
32
In recent years, co-extrusion of feed ingredients has been considered as a reasonable
option to improve the nutritional quality of soybean meal when used as a replacement
ingredient in laboratory manufactured shrimp feeds (Davis and Arnold, 2000; Mendoza et
al., 2001; Samocha et al., 2004). Results from this study are in agreement with findings
from these authors, in terms that commercial extrusion may have an important role in
improving the overall nutritional quality of shrimp diets including high levels of plant
protein sources. However, further studies are still required to evaluate the real benefits of
using previously co-extruded products as ingredients of shrimp diets that will be posteriorly
extruded.
Conclusion
Results from this study demonstrate that in commercially manufactured shrimp
feeds, fish meal can be completely removed from the formulation using alternative
vegetable protein sources in combination with poultry by-product meal without
compromising the production performance and economic returns of Litopenaeus vannamei
reared in pond systems. It is likely that the positive response of shrimp fed the replacement
feeds, was the result of a combination of adequate feed formulation and manufacture,
accurate feed inputs and management of the pond ecosystem and the contribution of natural
food organisms to the total feed intake. Further pond studies evaluating the potential of
Litopenaeus vannamei to utilize feeds without any animal proteins are suggested.
33
Aknowledgements
The authors would like to extend their thanks to those who have taken the time to
critically review this manuscripts as well as those who helped in supporting this research at
the Claude Peteet Mariculture Center and in Auburn University. This research was
supported in part by the United Soybean Board. Mention of a trademark or proprietary
product does not constitute an endorsement of the product by Auburn University and does
not imply its approval to the exclusion of other products that may also be suitable.
34
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Fox, J.M., Lawrence, A.L., Smith, F., 2004. Development of a low-fish meal feed
formulation for commercial production of Litopenaeus vannamei. Avances en
Nutricion Acuicola VII. Memorias del VII Simposium Internacional de Nutricion
Acuicola. 16-19 Noviembre, 2004. Hermosillo, Sonora, Mexico
Garza, A., Rouse, D.B., Davis, D.A., 2004. Influence of nursery period on the growth and
survival of Litopenaeus vannamei under pond production conditions. J. World
Aquac. Soc. 35 (3), 357-365.
Hardy, R.W., 1999. Alternate protein sources. Feed Management 50, 25-28.
Hernandez, C., Sarmiento-Pardo, J., Gonzalez-Rodriguez, B., Abdo de la Parra, I., 2004.
Replacement of fish meal with co-extruded wet tuna viscera and corn meal in diets
for white shrimp (Litopenaeus vannamei, Boone) Aquac. Res. 36, 834-840.
Hertrampf, J.W., Piedad-Pascual, F., 2000. Handbook on Ingredients for Aquaculture
Feeds. Kluwer Academic Publishers (ed.). Dordrecht, The Netherlands. 573 p.
Josupeit, H., 2004. An overview of the world shrimp market. World Shrimp Markets 2004,
26-27 October 2004, Madrid, Spain. http://www.globefish.org/
Lim, C., Dominy, W., 1990. Evaluation of soybean meal as a replacement for marine
animal protein in diets for shrimp (Penaeus vannamei). Aquaculture 87, 53-63.
Lim, C., Klesius, P.H., Dominy, W., 1998. Soyabean products. International Aqua Feeds 3,
17-23.
Mendoza, R., De Dios, A., Vazquez, C., Cruz, E., Ricque, D., Aguilera, C., Montemayor,
J., 2001. Fishmeal replacement with feather-enzymatic hydrolyzates co-extruded
with soya-bean meal in practical diets for the Pacific white shrimp (Litopenaeus
vannamei). Aquac. Nutr. 7, 143-151.
36
Moss, S.M., Divakaran, S., Kim, B.G., 2001. Stimulating effects of pond water on digestive
enzyme activity in the Pacific white shrimp, Litopenaeus vannamei (Boone). Aquac.
Res. 32, 125-131.
Moss, S.M., Pruder, G.D., 1995. Characterization of organic particles associated with rapid
growth in juvenile white shrimp, Penaeus vannamei (Boone), reared under intensive
culture conditions. J. Exp. Mar. Biol. Ecol. 187, 175-191.
NRC, National Research Council. 1993. Nutrient requirements of fish.
Samocha, T., Davis, D.A., Saoud, I.P., DeBault, K., 2004. Substitution of fish meal by co-
extruded soybean poultry by-product meal in practical diets for the Pacific white
shrimp, Litopenaeus vannamei. Aquaculture 231, 197-203.
Steel, R.G.D., Torrie, J.H., 1980. Principles and Procedures of Statistics: A Biometrics
Approach. McGraw-Hill, New York, NY, USA.
Swick, R.A., Akiyama, D.M., Boonyaratpalin, M., Creswell, D.C., 1995. Use of soybean
meal and synthetic methionine in shrimp feed. American Soybean Association,
Technical Bulletin (Vol. AQ43-1995).
Tacon A.G.J., 1996. Nutritional studies in crustaceans and the problems of applying
research findings to practical farming systems. Aquac. Nutr. 1, 165-174.
Tacon, A.G.J., Barg, U.C., 1998. Major challenges to feed development for marine and
diadromous finfish and crustacean species. In: De Silva, S.S. (Ed.), Tropical
Mariculture. Academic Press, San Diego, CA, USA, 171 - 208.
Tacon, A.G.J., Cody, J.J., Conquest, L.D., Divakaran, S., Forster, I.P., Decamp, O.E., 2002.
Effect of culture system on the nutrition and growth performance of Pacific white
shrimp Litopenaeus vannamei (Boone), fed different diets. Aquac. Nutr. 8, 121-137.
Venero, J., 2006. Optimization of dietary nutrient inputs for Pacific white shrimp
Litopenaeus vannamei. Doctoral dissertation. Auburn University, Auburn,
Alabama, USA. .
Zelaya, O., 2005. An evaluation of nursery techniques and feed management during culture
of marine shrimp Litopenaeus vannamei. Doctoral dissertation. Auburn University,
Auburn, Alabama, USA.
37
CHAPTER III
ALTERNATIVE DIETS FOR THE PACIFIC WHITE SHRIMP Litopenaeus vannamei
Abstract
Future use of animal protein sources in aquatic animal feeds is expected to be
considerably reduced as a consequence of increasing economical, environmental and safety
issues. Shrimp research has recently focused on the development of alternative feeds, with
minimal levels of marine protein sources. To determine shrimp capacity to use practical
feeds with varying levels of plant proteins, an 81-day growth trial was conducted using
juvenile (0.74 g) Litopenaeus vannamei stocked into replicated outdoor culture tanks.
Experimental treatments included four diets with varying levels of fish meal in the diet
(9%, 6%, 3% and 0%) in combination with 16% poultry by-product meal, a plant based
feed containing 1% squid meal, and a commercial reference feed. Feeds were commercially
extruded and offered as sinking pellets designed to contain 35% crude protein and 8%
lipids. Mean final weight, percent weight gain, final net yield, feed conversion ratio and
survival were evaluated at the end of the 81-day culture period. Final values for these
parameters ranged from 17.4 - 19.5 g, 2,249 - 2,465 %, 564.4 - 639.0 g/m3, 1.07 - 1.20 and
83.3 - 89.2 %, respectively. Evaluation of production parameters at the end of the study
demonstrated no significant differences (P> 0.05) among any of the experimental
38
treatments. These results indicate that fish meal can be replaced with plant protein sources
in shrimp diets including 16 % poultry by-product meal. Additionally, results demonstrate
the good performance can be obtained by using a combination of plant protein sources
(solvent extracted soybean meal, corn gluten meal and corn fermented solubles) in
combination with 1% squid meal. Although results with the primarily plant based diet are
encouraging, further evaluations are recommended to allow the removal of the remaining
marine ingredients.
Keywords: Litopenaeus vannamei, Soybean meal, Fish meal, Plant proteins.
39
Introduction
Animal origin ingredients such as fish meal, poultry by-product meal and meat and
bone meal are considered among the most suitable protein sources for shrimp feeds. In spite
of their importance, a considerable reduction in the use of these animal origin ingredients is
expected in coming years. Limited availability, variable supply and safety issues are
primary concerns. Given the growing demand by animal production industries for fish meal
and its limited supply, prices are likely to continue to increase, therefore, restraining future
its use as the main protein source in shrimp feeds.
Likewise, emerging environmental and safety issues associated to the use of
potentially contaminated animal by-products in animal feeds and the effect of fish meal
production from natural fish stocks have also been viewed negatively. It has been suggested
that a one way to address all these issues is through the development of all plant-protein
feeds. Such a tactic could also provide an economical opportunity for shrimp producers, as
some segments of the market would pay a higher price for a premium shrimp fed and
produced under environmentally sound conditions (Josupeit, 2004; Davis et al., 2004;
Samocha et al., 2004).
Facing these scenarios, various studies have focused on the development of
alternatives that effectively replace or minimize the inclusion of animal protein sources in
commercial shrimp formulations using plant proteins (Colvin and Brand, 1977; Lim and
Dominy, 1990, Piedad-Pascual et al., 1990; Hardy, 1999; Divakaran et al., 2000; Davis and
Arnold, 2000; Conklin, 2003; Samocha et al., 2004). According to Davis et al. (2004), the
40
use of an all-plant protein feed can be limited due to a variety of factors including,
deficiency or imbalance of essential amino acids, reduced levels of minerals, limited levels
of highly unsaturated fatty acids (HUFA), presence of anti-nutritional factors or toxins and
decreased palatability. In spite of these limitations, and that nutritional information on
shrimp is far from complete, the understanding of primary nutrient requirements for shrimp
is adequate to allow the replacement of animal protein sources with alternative ingredients..
It is clear then, that the complete substitution of fish meal and animal by-product
meals with other protein sources can be effectively achieved only when certain basic
conditions are provided. Of primary concern is the proper formulation and supplementation
of feeds with adequate lipids, phosphorus and amino-acids sources (i.e. lysine and
methionine), to overcome the nutritional imbalances that arise when marine animal meals
are removed from the formulation. (Akiyama,1988; Akiyama, 1991; Akiyama et al., 1991).
Furthermore, proper feed manufacture and feedstuffs processing have been found to
improve the overall nutritional quality and water stability of replacement feeds (Davis and
Arnold, 2000; Hernandez et al., 2004; Samocha et al., 2004). According to Carver et al.
(1989), extrusion has the advantage of inactivating and/or destroying some of the heat-
sensitive anti-nutritional factors found in plant protein sources, such as soybean meal, while
also gelatinizing starch granules.
Just as the nutritional component of the feed is of primary importance in shrimp
nutrition, feeding habits also play an important role when considering the implementation
of feeds with minimal animal origin ingredients. Shrimp, for example, have the capacity to
utilize naturally available foods in green water systems where they are commonly cultured.
41
This means that shrimp growth in ponds is achieved through the simultaneous consumption
of feed and endogenously produced food organisms such as micro-algae and microbial-
detrital aggregates (Moss et al., 1995; Moss and Pruder, 1995 Moss et al, 2001; Forster et
al., 2002; Tacon et al., 2002). Studies evaluating the ability of shrimp to utilize diets with
low levels of animal protein sources have been commonly carried out in clear water
systems, where the length of the culture period is limited and environmental conditions
greatly differ from those found in commercial ponds (D?abramo and Castell, 1997). In
addition, these studies have generally used laboratory made feeds that are appropriate for
laboratory-scale studies, but lack the manufacturing and extrusion advantages provided by
commercial feeds.
There is limited information with regards to shrimp performance when fed extruded
feeds with low levels of animal protein sources and reared in green water systems. Hence,
the aim of this study was to evaluate the productive performance of the Pacific white
shrimp (Litopenaeus vannamei) fed a commercially manufactured feed with varying levels
of animal and vegetable protein sources and reared in an outdoor, semi-closed, recirculating
system in which the shrimp had access to natural food.
Materials and Methods
Shrimp and experimental units
Juvenile Pacific white shrimp (Litopenaeus vannamei) were obtained from 16
shrimp production ponds located at the Claude Peteet Mariculture Center, in Gulf Shores,
42
Alabama. Shrimp (mean weight + SD, 0.743 + 0.031g) were stocked at a density of 37.5
shrimp/m3 (30 shrimp/tank) in an outdoor, semi-closed recirculating system. The system
was composed by 24, circular polyethylene tanks (0.85m height x 1.22 m upper diameter,
1.04 m lower diameter) designed to contain 800-L each and a reservoir tank (800-L) with a
biological filter. Before stocking, the system was filled with brackish green water (9.50 ppt)
from a shrimp production pond. Daily water exchanges were performed during the
experimental period, between 4:00 am and noon, when pond water was pumped into the
central filter at a rate of 8 liters per minute. This exchange rate allowed a 100% water
exchange in the recirculating system every six days. Aeration was provided in the filter and
in each tank by two air stones connected to a common air supply from a 1-hp regenerative
blower.
During the experimental period, dissolved oxygen (DO), temperature, salinity and
pH concentrations were measured in the central filter and two of the system tanks at 06:00
in the morning and at 16:00 in the afternoon. Water quality parameters were measured
using a YSI 556MPS meter (Yellow Spring Instrument Co., Yellow Springs, OH, USA).
Total ammonia-nitrogen (TAN) was determined on a weekly basis, with water samples
taken from the filter and two of the tanks and measured with a spectrophotometer
(Spectronic Instrument Inc. Rochester, NY, USA) using the Nesslerization method (APHA
1989).
43
Feeds and feed management
The six dietary treatments (Table 1) were randomly assigned to shrimp in four
replicate tanks per treatment. The goal for the formulation of the first four diets, was to
reduce the inclusion of menhaden fish meal (FM) as follows: 9%, 6%, 3% and 0% FM of
the total diet, while increasing the inclusion of vegetable protein sources, mainly solvent
extracted soybean meal and corn gluten meal.
Poultry by-product meal (PBM) was fixed at 16% inclusion in the first four diets.
The fifth diet.was a second generation plant protein based feed, formulated using the profile
of the diet previously reported by Davis et al. (2004), but including 1% squid meal as an
attractant and source of essential nutrients. Because this plant protein feed was also
formulated to be used in the low salinity conditions of west Alabama waters, it included an
additional supplementation of 2% of potassium chloride. The sixth diet was a 35% protein
commercial shrimp feed manufactured by Rangen Inc. and served as a high quality
commercial reference. The dietary treatments were formulated in order to provide equal
protein and lipid levels while maintaining a minimum lysine and methionine+cystine
content of 5% and 3% of the total protein, respectively (Table 2). Corn gluten meal was
used as a natural source of methionine and no crystalline amino acids were supplemented.
Corn fermented solubles were used as an additional protein source and only added to the
plant protein based diet.
44
Table 1. Ingredient composition of practical diets for Litopenaeus vannamei used to
evaluate the replacement of animal proteins by plant protein sources (values expressed on
an as fed basis, g/100 g). Diets were commercially manufactured by Ranger? Inc.
(Angleton, TX) using extrusion processing.
Ingredient 9% FM 6% FM 3% FM 0% FM Plant Based
Soybean Meal 32.48 34.82 37.17 39.52 56.46
Fish Meal-Menhaden 9.00 6.00 3.00 - -
Poultry By-product meal 16.00 16.00 16.00 16.00 -
Milo 35.47 33.82 32.33 30.68 14.99
Corn Gluten - 1.67 3.17 4.84 4.83
Fish Oil 3.96 4.22 4.47 4.72 5.76
Di-calcium Phosphate 1.50 1.88 2.27 2.65 3.38
Corn Fermented Solubles - - - - 9.99
Salt-Potassium chloride - - - - 2.00
Squid Meal - - - - 1.00
Bentonite 1.00 1.00 1.00 1.00 1.00
Mold Inhibitor 0.15 0.15 0.15 0.15 0.15
Vitamin Premix 0.34 0.34 0.34 0.34 0.34
Mineral Premix 0.08 0.08 0.08 0.08 0.08
Stay-C (35% Active Vit. C) 0.02 0.02 0.02 0.02 0.02
45
Table 2. Nutritional composition of practical diets for Litopenaeus vannamei used to
evaluate the replacement of animal proteins with plant protein sources1.
Nutrient 9% FM 6% FM 3% FM 0% FM Plant
Based
Reference
Crude Protein 35.7 35.9 36.2 36.6 36.1 37.6
Crude Fat 8.4 8.3 8.6 8.4 8.4 10.6
Crude Fiber 2.4 1.8 2.1 1.9 2.2 1.7
Ash 8.2 7.9 7.9 8.1 9.5 8.9
Calcium2 1.3 1.3 1.2 1.1 0.9 -
Total Phosphorus2 1.2 1.2 1.2 1.3 1.2 -
Lysine2 2.0 2.0 1.9 1.8 2.0 -
Met + Cys2 1.1 1.1 1.1 1.1 1.1 -
% Protein from plant sources2,3 54.9 60.4 66.0 71.5 97.7 -
% Protein from animal sources2,4 45.1 39.6 34.0 28.5 2.3 -
1 Analyzed value - New Jersey Feed Laboratory, Inc. Trenton, NJ.
2 Formulated value.
3 Soybean meal, corn gluten meal, milo, corn fermented solubles.
4 Fish meal, poultry by-product meal.
5 Squid meal.
46
Lipid levels were adjusted by adding menhaden fish oil, which is a rich source of
highly unsaturated fatty acid (HUFA), such as decoeicosanoic acid (DHA) and arachidonic
acid (AA). Calcium phosphate was added to ensure an adequate phosphorus supply. Feeds
were produced by Rangen Inc. (Angleton, TX. USA) under commercial manufacturing
conditions and offered to the shrimp as a sinking extruded pellet.
Daily feed inputs were back-calculated, based upon an expected shrimp weight gain
of 1 g per week and an expected feed conversion ratio of 1.8:1. Feed was provided twice
per day. In order to readjust daily feed inputs, shrimp survival was determined by counting
all shrimp in each tank on a bi-weekly basis. Overall, each shrimp in each tank was offered
a total of 20 g of feed throughout the 81-day experimental period. Feed inputs were stopped
one day before harvesting. Shrimp production was evaluated at the end of the growth trial
considering the following parameters: mean final weight, weight gain (%), weekly weight
gain, final net yield, feed conversion ratio (FCR) and survival. In order to evaluate the
economic feasability of using alternative shrimp diets, the following parameters were also
determined: total value of shrimp production, feed price, cost for total feed inputs, partial
?gross? returns and feed cost per kilogram of shrimp produced.
Statistical Analyses
Data were analyzed by using one way analysis of variance (ANOVA), to determine
if significant (p<0.05) differences existed among treatment means. The Student?Neuman
?Keuls multiple comparison test was used to determine whether significant differences
47
existed between treatment means (Steel and Torrie, 1980). All statistical analyses were
conducted using SAS (V9.1., SAS Institute, Cary, NC, USA).
Results
Water quality parameters throughout the 81-day growth trial were maintained at
suitable levels for adequate growth and survival of shrimp (Table 3). Mean final weight,
weight gain (%), final net yield, FCR and survival were evaluated at the end of the 81-day
culture period. Final values for these parameters ranged from 17.4 - 19.5 g, 2,249 - 2,465
%, 564.4 - 639.0 g/m3, 1.07 - 1.20 and 83.3 - 89.2 %, respectively. Complete data of
productivity and economic parameters of Litopenaeus vannamei fed the six dietary
treatments are presented in Tables 4 and 5. Results indicate that none of the parameters
evaluated were significantly affected by the inclusion of plant protein sources as a
replacement for fish meal and poultry by-product meal for any of the dietary treatments.
Although not statistically significant differences were found among any of the treatments
and productive parameters evaluated, shrimp fed the 0% FM diet numerically showed the
poorest performance for most parameters evaluated, except for survival. Experimental units
offered the plant protein based diet showed the lowest shrimp survival. Final mean weight
of shrimp fed the reference feed (19.5 g) was numerically higher than those of shrimp fed
the other treatments, with the lowest mean value for shrimp fed the 0% FM feed (17.4 g).
Lowest mean weight gain (%) was found in the 0% FM (2,249%) treatment. Mean final net
yield ranged from 0.56 kg/m3 (0% FM) to 0.64 kg/m3 (Reference diet) and was within the
range of yields given for intensive shrimp production systems.
48
Table 3. Summary of water quality parameters observed over an 81-day experimental
period for Litopenaeus vannamei fed practical diets with varying levels of animal and plant
protein sources and cultured in an outdoor semi-closed recirculating culture system. Values
are mean ? standard deviation of daily and weekly determinations. Values in parenthesis
represent minimum and maximum readings throughout the study.
Temperature
(C)
DO1
(mg/l)
pH Salinity
(ppt)
TAN
(mg/l)2
am 27.4 ?1.3
(24.1, 29.7)
6.70 ? 0.56
(3.79, 7.67)
7.30 ? 0.28
(6.24, 8.18)
8.3 ? 0.8
(7.0, 9.5)
0.31 ? 0.35
(0.00, 0.88)
pm 29.6 ? 1.8
(23.9, 32.8)
6.66 ? 0.54
(4.70, 7.85)
7.66 ? 0.25
(7.17, 8.29)
1 Dissolved Oxygen.
2 Total Ammonium Nitrogen. Weekly values.
49
Table 4. Mean production parameters at the end of an 81-day culture period for Litopenaeus
vannamei reared in an outdoor semi-closed recirculating culture system and fed practical
diets with varying levels of fish meal (FM), a plant based diet and a commercial reference
diet1.
Parameter 9% FM 6% FM 3% FM 0% FM Plant Ref. PSE 2 P-value
Initial weight (g) 0.72 0.73 0.75 0.74 0.72 0.78 0.014 0.056
Final mean weight (g) 18.5 18.6 18.8 17.4 18.4 19.5 0.439 0.072
Weight gain (%)3 2,443 2,439 2,391 2,249 2,465 2,401 80.742 0.486
Weekly weight gain (g) 1.53 1.55 1.56 1.44 1.53 1.61 0.038 0.096
Final net yield (g/m3) 586.2 624.0 622.5 564.4 574.0 639.0 18.752 0.055
FCR4 1.13 1.12 1.11 1.20 1.13 1.07 0.030 0.108
Survival (%) 85.0 89.2 88.3 86.7 83.3 87.5 2.766 0.687
1 Based on analysis of variance (ANOVA), no significant differences (P > 0.05) were found
among treatment means (n = 4).
2 Pooled standard error of treatment means = .mse n
3 Weight gain (%) = 100 x (final weight - initial weight) / initial weight.
4 FCR, feed conversion ratio = Feed offered per shrimp / weight gain per shrimp.
50
Table 5. Mean economic parameters of Litopenaeus vannamei cultured in an outdoor semi-
closed recirculating system over an 81-day experimental period and fed practical diets with
varying levels of fish meal (FM) and a plant protein based diet. Estimates are based on
production per cubic meter of culture system1.
Parameter 9% FM 6% FM 3% FM 0% FM Plant P-Value PSE
Shrimp production (g/m3) 586.2 624.0 622.5 564.4 574.0 0.150 19.6
Production value (USD)2 2.12 2.26 2.26 2.05 2.08 0.149 0.07
Total feed inputs (g/m3) 637.5 668.7 662.5 650.0 625.0 0.630 22.1
Feed price (USD/kg)3 0.531 0.526 0.520 0.515 0.532 - -
Total feed inputs cost (USD)4 0.34 0.35 0.34 0.33 0.33 0.778 0.01
Partial gross returns (USD)5 1.79 1.91 1.92 1.71 1.75 0.120 0.06
Feed cost per kg of shrimp
produced (USD)6
0.60 0.59 0.58 0.62 0.60 0.308 0.015
1 Based on analysis of variance (ANOVA) no significant differences (P > 0.05) were found
among treatment means (n = 4). Reference feed excluded from the economic analysis.
2 Production Value = Shrimp production per m3 x $3.63 USD/Kg of shrimp. Shrimp price is
based on Gulf fresh shrimp prices (head-on) between the 13 and 19 of October, 2005, as
reported by the National Marine Fisheries Service, Fisheries Statistics Division, Silver
Spring, MD,USA .
3 Price on May 2005. Subject to change based on varying ingredients price. Prices were
based on those at the time of the research. As fish meal has gone up in price and prices vary
from region to region the cost effectiveness of the feed will change. Hence, conclusion with
regards to cost effectiveness should be made on a case by case basis.
4 Total feed inputs cost = Total Feed Inputs (g/m3) x Feed price (USD/kg).
5 Partial Gross Returns = Production value (USD) - Total Feed Cost (USD/kg).
6 Feed cost per kg of shrimp produced = FCR x Feed price (USD/kg).
51
Feed conversion ratio (FCR) and survival values from this study ranged between
1.11 (3% FM) to 1.20 (0% FM ) and 83.3% (0% FM) to 89.2% (6% FM), respectively. Low
P-values of 0.056 and 0.055 were found for initial weight at stocking and final shrimp
production (g/m3) analyses, respectively, whereas weight gain (%), FCR and survival
analyses resulted in larger P-values of 0.486, 0.108 and 0.687 respectively. No statistical
significant differences were found among treatment means for any of these parameters.
Feed prices showed a slight decrease of nearly sixteen dollars between the 9% FM
feed ($531/mt feed) and the 0% FM feed ($515/mt feed). There was a general trend toward
reduced price as more plant protein sources were included in the formulation. Conversely,
the plant protein diet showed a higher price than all of the other treatments ($532/mt feed).
Higher cost of the plant protein diet was the consequence of extra costs associated with the
purchase of small quantities of select ingredients (e.g. corn fermented solubles), which is
not a typical ingredient at this mill.
Discussion
Results from this study provide important information regarding the potential of
Litopenaeus vannamei to utilize alternative feed formulations under commercial production
conditions. This research was conducted in replicated outdoor tanks using green water from
a production pond and the shrimp were fed commercially extruded diets. Because of these
production characteristics, the rearing conditions used in this study closely resembled those
found by shrimp in commercial semi-intensive production systems. Therefore, shrimp
utilization of the diets used in this study are indicative of the important potential of
52
Litopenaeus vannamei to use similar diets under commercial pond conditions. The good
production results observed in this study confirm that Litopenaeus vannamei can be fed
commercially manufactured feeds including plant proteins as replacements of animal
protein sources, without adverse effects on shrimp performance or production economics.
Shrimp performance was excellent across all treatments with no statistically
significant differences among the diets. Production parameters were similar to those of
shrimp produced using similar management procedures and reared in outdoor recirculating
systems in tanks (Venero, 2006; Davis et al., 2004; Samocha et al., 2004). The numerically
lower growth of shrimp fed the 0% FM feed, may indicate that further optimization of the
diet may be possible. The plant diet which included 1% squid meal and resulted in the
highest shrimp weight gain among all treatments (2,465%) also showed the lowest survival.
This increased growth could have been influenced by the reduced density. Although these
results are not significantly different, further evaluations and refining of plant-based feed
formulations for shrimp are recommended.
Findings from this study are in agreement with those of Davis et al. (2004), who
using a similar rearing system, reported the successful replacement of animal protein
sources with plant proteins in shrimp feeds containing fish oil. These authors also claimed
that both fish meal and marine oil sources could be removed from shrimp feeds if suitable
alternative sources of protein and lipids are provided to meet essential amino acid and fatty
acid requirements of the shrimp.
Besides the nutritional component of the feed, part of the success of the replacement
of animal proteins by alternative sources is supported on the shrimp capacity to utilize
53
natural productivity. Different authors have indicated that the enhanced growth and
performance of animals reared under ?green-water? culture conditions is due to their ability
to obtain additional nutrients from natural food organisms present in green water systems
and/or pond ecosystems (Leber and Pruder 1988; Moss 1995; Tacon 1996; Moriarty 1997).
Shrimp benefit from a wide range of organisms within the green water culture environment,
including bacteria, phytoplankton, protists, rotifers and nematodes (Moss and Pruder, 1995;
Schuur, 2003; Decamp et al., 2003). For example, among the natural food organisms some
flagellates and ciliates, have been reported to be rich in highly unsaturated fatty acids (i.e.
16-25% of the total) which have a good growth promoting effect on juvenile Litopenaeus
vannamei (Lim et al., 1997). The importance of this endogenous community in intensive
shrimp production systems, was reported by Gomez-Jimenez et al. (2005), who did not find
differences in weight gain, final weight or survival of Litopenaeus vannamei offered
commercial diets with varying levels of protein (25%, 30%, 35% or 40%CP) in a zero
water exchange system.
Even when some productive advantages are acquired with the use of these kind of
systems, to capitalize on potential markets for shrimp grown under organic or
environmentally sustainable conditions, one must also utilize organic ingredients from
more sustainable sources (Davis et al., 2004). Since one of the premises of sustainable
aquaculture is to minimize the use of resources of limited availability, further studies
evaluating the replacement of the fish oil from these diets are recommended.
54
Conclusion
This study demonstrates that fish meal can be removed from commercially
manufactured shrimp diets including 16% poultry by-product meal using vegetable protein
sources, with no adverse effect on the productive performance of Litopenaeus vannamei
reared in green water environments. Furthermore this study provided important evidence
that animal protein sources can be completely removed from shrimp feeds without
negatively affecting shrimp growth. Shrimp feeds in this study included varying levels of
soybean meal, corn gluten meal and corn fermented solubles as alternative protein sources
to fish meal and poultry by-product meal. Although the all-plant diet produced good shrimp
performance, this diet may be marginal and further studies are recommended to evaluate
potential limiting nutrients as well as the efficacy of squid meal. Additional studies with
plant based diets at a larger scale under commercial pond conditions are also suggested.
Aknowledgements
The authors would like to extend their thanks to those who have taken the time to
critically review this manuscripts as well as those who helped support this research at the
Claude Peteet Mariculture Center and at Auburn University. This research was supported in
part by the United Soybean Board. Mention of a trademark or proprietary product does not
constitute an endorsement of the product by Auburn University and does not imply its
approval to the exclusion of other products that may also be suitable.
55
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and R.KH. Tan) Sept. 19-25. American Soybean Association, Singapore. pp. 80-90.
APHA (American Public Health Association), American Water Works Association, and
Water Pollution Control Association. 1989. Standard Methods for the Examination
of Water and Waste Water, 17th edition. American Public Health Association,
Washington, D.C., USA.
Carver, L.A., Akiyama, D.M., Dominy, W.G., 1989. Processing of wet shrimp heads and
squid viscera with soy meal by a dry extrusion process. American Soybean
Association Technical Bulletin. 3, 777 Craig Road, St. Louis, MO, USA. AQ16, 89-
4.
Colvin, L.V., Brand, C.W., 1977. The protein requirement of penaeid shrimp at various
life-cycle stages in controlled environment systems. Proc. World Maric. Soc. 8,
821-840.
Conklin, D.E., 2003. Use of Soybean Meal in the Diets of Marine Shrimp. Department of
Animal Science, University of California, Davis, United Soybean Board and
American Soybean Association. Technical review paper AQ 144-2003.
D'Abramo, L.R., Castell, J.D., 1997. Research methodology. In: Crustacean Nutrition,
Advances in World Aquaculture (ed. by L.R. D'Abramo D.E. Conklin and D.M.
Akiyama), pp. 3-25. World Aquaculture Society, Louisiana.
Davis, D.A., Arnold. C.R., 2000. Replacement of fish meal in practical diets for the Pacific
white shrimp, Litopenaeus vannamei. Aquaculture185, 291-298.
Davis, D.A.,Samocha, T.M., Bullis, R.A., Patnaik, S., Browdy, C., Stokes, A. and Atwood,
H., 2004. Practical Diets for Litopenaeus vannamei, (Boone. 1931): Working
Towards Organic and/or All Plant Production Diets. Avances en Nutricion Acuicola
VII. Memorias del VII Simposium Internacional de Nutricion Acuicola. 16-19
Noviembre, 2004. Hermosillo, Sonora, Mexico
56
Decamp, O., Cody, J., Conquest, L., Delanoy, G., Tacon, A.G.J., 2003. Effect of salinity on
natural community and production of Litopenaeus vannamei, (Boone), within
experimental zero-water exchange culture systems. Aquac. Res. 34, 345-355.
Dersjant-Li, Y., 2002. The use of soy protein in aquafeeds. Avances en Nutricion Acuicola
VI. Memorias del VI Simposium Internacional de Nutricion Acuicola. 3-6 de
Septiembre del2002. Cancun, Quintana Roo, Mexico.
Divakaran, S., Velasco, M., Beyer, E., Forster, I., Tacon, A.G.J., 2000. Soybean meal
apparent digestibility for Litopenaeus vannamei, including a critique of
methodology. Avances en Nutricion Acuicola V. Memorias del V Simposium
Internacional de Nutricion Acuicola. 19-22 Noviembre, 2000. Merida, Yucatan,
Mexico.
Forster, I.P., Dominy, W., Tacon, A.G.J., 2002. The use of concentrates and other soy
products in shrimp feeds. Avances en Nutricion Acuicola VI. Memorias del VI
Simposium Internacional de Nutricion Acuicola. 3-6 de Septiembre del 2002.
Cancun, Quintana Roo, Mexico.
Fox. J.M., Lawrence, A.L., Smith, F., 2004. Development of a low-fish meal feed
formulation for commercial production of Litopenaeus vannamei. Avances en
Nutricion Acuicola VII. Memorias del VII Simposium Internacional de Nutricion
Acuicola. 16-19 Noviembre, 2004. Hermosillo, Sonora, Mexico
Gomez-Jimenez, S., Gonzalez-Felix, M.L., Perez-Velazquez, M., Trujillo-Villalba, D.A.,
Esquerra-Brauer, I.R., Barraza-Guardado, R., 2005. Effect of dietary protein level
on growth, survival and ammonia efflux rate of Litopenaeus vannamei (Boone)
raised in a zero water exchange culture system. Aquac. Res. 36, 834-840.
Gonzalez-Rodriguez, B., Abdo de la Parra, I., 2004 Replacement of fish meal with co-
extruded wet tuna viscera and corn meal in diets for white shrimp (Litopenaeus
vannamei, Boone). Aquac. Res. 35, 1153-1157.
Hardy, R.W., 1999. Alternate protein sources. Feed Management 50, 25-28.
Hernandez, C., Sarmiento-Pardo, J., Gonzalez-Rodriguez, B., Abdo de la Parra, I., 2004.
Replacement of fish meal with co-extruded wet tuna viscera and corn meal in diets
for white shrimp (Litopenaeus vannamei, Boone) Aquac. Res. 36, 834-840.
Hertrampf, J.W., Piedad-Pascual, F., 2000. Handbook on Ingredients for Aquaculture
Feeds. Kluwer Academic Publishers (ed.). Dordrecht, The Netherlands. 573 p.
Josupeit, H., 2004. An overview of the world shrimp market. World Shrimp Markets 2004,
26-27 October 2004, Madrid, Spain. http://www.globefish.org/
57
Leber, K.M., Pruder, G.D., 1988. Using experimental microcosms in shrimp research: the
growth-enhancing effect of shrimp pond water. J. World Aquac. Soc. 19, 197-203.
Lim, C., Dominy, W., 1990. Evaluation of soybean meal as a replacement for marine
animal protein in diets for shrimp (Penaeus vannamei). Aquaculture 87, 53-63.
Lim, C., Ako, H., Brown, C.L., Hahn, K. 1997. Growth response and fatty acid composition
of juvenile Pennaeus vannamei fed different sources of dietary lipid. Aquaculture
151, 143-153.
Lim, C., Klesius, P.H., Dominy, W., 1998. Soyabean products. International Aqua Feeds 3,
17-23.
Mendoza, R., De Dios, A., Vazquez, C., Cruz, E., Ricque, D., Aguilera, C., Montemayor,
J., 2001. Fishmeal replacement with feather-enzymatic hydrolyzates co-extruded
with soya-bean meal in practical diets for the Pacific white shrimp (Litopenaeus
vannamei). Aquac. Nutr. 7, 143-151
Moriarty, D.J.W., 1997. The role of microorganisms in aquaculture ponds. Aquaculture
151, 333-349.
Moss, S.M., Divakaran, S., Kim, B.G., 2001. Stimulating effects of pond water on digestive
enzyme activity in the Pacific white shrimp, Litopenaeus vannamei (Boone). Aquac.
Res. 32, 125-131
Moss, S.M., Pruder, G.D., 1995. Characterization of organic particles associated with rapid
growth in juvenile white shrimp, Penaeus vannamei Boone, reared under intensive
culture conditions. J. Exp. Mar. Biol. Ecol. 187, 175-191.
Piedad-Pascual, F., Cruz, E.M., Sumalangcay, A., 1990. Supplemental feeding on Penaeus
monodon juveniles with diets containing various levels of defatted soybean meal.
Aquaculture, 89: 183-191.
Samocha, T., Davis, D.A., Saoud, I.P., DeBault, K., 2004. Substitution of fish meal by co-
extruded soybean poultry by-product meal in practical diets for the Pacific white
shrimp, Litopenaeus vannamei. Aquaculture 231, 197-203.
Schuur A.M., 2003. Evaluation of biosecurity applications for intensive shrimp farming.
Aquac. Eng. 28, 3-19.
Steel, R.G.D., Torrie, J.H., 1980. Principles and Procedures of Statistics: A Biometrics
Approach. McGraw-Hill, New York, NY, USA.
58
Swick, R.A., Akiyama, D.M., Boonyaratpalin, M., Creswell, D.C., 1995. Use of soybean
meal and synthetic methionine in shrimp feed. American Soybean Association,
Technical Bulletin (Vol. AQ43-1995).
Tacon A.G.J., 1996. Nutritional studies in crustaceans and the problems of applying
research findings to practical farming systems. Aquac. Nutr. 1, 165-174.
Tacon, A.G.J., Barg, U.C., 1998. Major challenges to feed development for marine and
diadromous finfish and crustacean species. In: De Silva, S.S. (Ed.), Tropical
Mariculture. Academic Press, San Diego, CA, USA, 171 - 208.
Tacon, A.G.J., Cody, J.J., Conquest, L.D., Divakaran, S., Forster, I.P., Decamp, O.E., 2002.
Effect of culture system on the nutrition and growth performance of Pacific white
shrimp Litopenaeus vannamei (Boone) fed different diets. Aquac. Nutr. 8, 121-137.
Venero, J., 2006. Optimization of dietary nutrient inputs for Pacific white shrimp
Litopenaeus vannamei. Doctoral dissertation. Auburn University, Auburn,
Alabama, USA.
59
IV. SUMMARY AND CONCLUSIONS
The primary objective of this study was to evaluate the replacement of fish meal and
other animal proteins by using alternative plant protein sources in commercially
manufactured diets for Litopenaeus vannamei reared under green water conditions.
Currently there is pressure to minimize the use of animal protein sources, mainly fish meal,
in shrimp feeds. Limited fish meal supply and high demand for this ingredient by other
animal productions make fish meal one of the primary ingredient costs in shrimp
formulations. Regardless of this phenomena, commercial formulations have been
commonly reported to contain between 25 - 50 % of fish meal of the total diet. With shrimp
market prices not growing in value in recent years and feed representing the main cost
associated with the production of shrimp, the evaluation of nutritional strategies focusing
on replacing fish meal with cheaper protein sources seems a reasonable option to contribute
to the reduction of shrimp production costs
The use of plant protein sources as alternative ingredients to fish meal have
important ecological and economical implications. For example, reduced fish meal levels in
shrimp diets would help minimize pressure on limited fish meal resources with the added
advantage of helping to reduce the cost of the feeds as higher levels of cheaper plant protein
sources are incorporated into the diets. In addition, alternative feeds (e.g. using organically-
grown ingredients) will allow producers to reach higher value markets where consumers
60
may be willing to pay a premium price for shrimp fed diets with no animal origin
ingredients and produced using technologies and practices that represent a minimal threat to
the environment or the human health. Shrimp do not have a specific requirement for
ingredients of animal origin in their diet, rather, shrimp need to be supplied a basic level of
nutrients to effectively fulfill their biological demands for adequate maintenance, growth
and/or reproduction. Consequently, the key to the effective use of plant-based feeds by
shrimp is to have an adequate knowledge of the nutritional requirements of the animal and
a good understanding of limiting factors associated with the use of plant ingredients in
practical diets. Besides the nutritional component, adequate ingredient processing and feed
manufacturing also play a key role to further increase the use of plant protein sources in
shrimp feeds. Properly processed, plant ingredients are characterized by having a low
content of anti-nutritional factors and high digestibility and protein content, making them
more suitable as a replacement for fish meal.
Based on these principles for the use of alternative shrimp diets and considering that
the practical application of studies conducted in controlled, clear water conditions is
limited, two studies were carried out in semi-commercial shrimp culture conditions to
evaluate Litopenaeus vannamei potential to use alternative plant based diets in production
systems with access to natural food organisms. The first study evaluated the performance of
shrimp fed four practical diets with varying levels of fish meal (9, 6, 3, 0 %) and plant
protein sources (soybean meal and corn gluten meal). All of the feeds evaluated in this
study included poultry by-product meal at a fixed level of 16% of the diet. In this study, a
minimal water exchange ponds production system was used to raise shrimp to commercial
61
size over an 18-week experimental period. In the second study, the same four diets from the
first study were evaluated in addition to a practical plant protein-based diet with no poultry
by-product meal, and an additional commercial reference diet. In this study, shrimp were
reared to commercial size in a green water semi-closed recirculating system, over an 81-day
culture period.
Findings from these two studies demonstrated that in commercially manufactured
shrimp diets including poultry by-product meal, fish meal can be completely replaced using
alternative protein sources, such as soybean meal and corn gluten meal, without negatively
compromising the productive performance and economic returns of Litopenaeus vannamei
reared in practical shrimp production systems. Results from these studies also provided
evidence that the cost of commercial shrimp diets can be reduced as increasing levels of
plant sources are included in the diets as replacement ingredients to fish meal.
In addition to these findings, the second study also provided interesting information
about the important potential of Litopenaeus vannamei to effectively use alternative plant-
based diets. In this study, shrimp fed the plant-based diet showed no significant differences
in their productive performance as compared to shrimp fed the diets including fish meal
and/or poultry by-product meal and shrimp fed the commercial reference diet. These are
encouraging findings; however, further work refining and improving the plant based diet
must be carried out in order to remove squid meal, and ensure the nutritional adequacy of
this diet.
62
It is likely that the positive response of shrimp fed the replacement feeds was the
result of a combination of different elements, such as adequate feed formulation and
manufacture, accurate feed inputs and management of the pond and tanks ecosystem and
the contribution of natural food organisms to the total nutrient intake. Further studies
evaluating the potential of Litopenaeus vannamei to utilize plant-based diets without any
poultry by-product meal and squid meal are suggested.
Results from this study demonstrate that Litopenaeus vannamei reared in green
water environments can use plant protein sources as replacement ingredients to fish meal in
commercially manufactured shrimp diets without negatively affecting production while
reducing feed costs. Based on the findings from this study, it is recommended to use plant
proteins, such as solvent extracted soybean meal and corn gluten meal as effective
alternative ingredients to fish meal and poultry by-product meal in shrimp feeds. These
kind of ingredients can be successfully used without affecting animal performance in
commercial shrimp production.
The most important implications from these findings are associated to the fact that
current fish meal inclusion levels can be considerably minimized, up to a 0% of inclusion,
using alternative plant protein ingredients in commercial shrimp diets. Increasing use of
cheaper plant proteins will allow for a important reduction on the cost of the feeds as costly
marine ingredients are removed from the formulations, or at least used at more efficient
levels. In addition, the optimization on the use of fish meal in shrimp feeds will also result
in a reduction in the demand for this limited ingredient while contributing to reduce the
pressure on wild fish stocks, used for the production of fish meal. Furthermore, the
63
possibility of reaching higher value markets that may help in improving profitability for
shrimp producers will also be favored by the use of organic feeds not including animal
origin ingredients. Although the performance of shrimp fed the plant-based diet in this
study was good, there may be limitations of specific nutrients as well as palatability
problems that should be further evaluated.
64
V. LITERATURE CITED
Akiyama, D.M., 1988. Soybean meal utilization by marine shrimp. AOCS world congress
on vegetable protein utilization in human food and animal feedstuffs, Singapore,
October 2-7. American Soybean Association, Singapore.
Akiyama, D.M., Dominy, W.G., Lawrence, A.L., 1991. Penaeid shrimp nutrition for the
commercial feed industry revised. In: Proceedings of the Aquaculture Feed
Processing and Nutrition Workshop. Thailand and Indonesia,(ed. by D.M. Akiyama
and R.KH. Tan) Sept. 19-25. American Soybean Association, Singapore. pp. 80-90.
APHA (American Public Health Association), American Water Works Association, and
Water Pollution Control Association. 1989. Standard Methods for the Examination
of Water and Waste Water, 17th edition. American Public Health Association,
Washington, D.C., USA.
Carver, L.A., Akiyama, D.M., Dominy, W.G., 1989. Processing of wet shrimp heads and
squid viscera with soy meal by a dry extrusion process. American Soybean
Association Technical Bulletin. 3, 777 Craig Road, St. Louis, MO, USA. AQ16, 89-
4.
Colvin, L.V., Brand, C.W., 1977. The protein requirement of penaeid shrimp at various
life-cycle stages in controlled environment systems. Proc. World Maric. Soc. 8,
821-840.
Conklin, D.E., 2003. Use of Soybean Meal in the Diets of Marine Shrimp. Department of
Animal Science, University of California, Davis, United Soybean Board and
American Soybean Association. Technical review paper AQ 144-2003.
D'Abramo, L.R., Castell, J.D., 1997. Research methodology. In: Crustacean Nutrition,
Advances in World Aquaculture (ed. by L.R. D'Abramo D.E. Conklin and D.M.
Akiyama), pp. 3-25. World Aquaculture Society, Louisiana.
Davis, D.A., Arnold, C.R., 2000. Replacement of fish meal in practical diets for the Pacific
white shrimp, Litopenaeus vannamei. Aquaculture185, 291-298.
65
Davis, D.A.,Samocha, T.M., Bullis, R.A., Patnaik, S., Browdy, C., Stokes, A. and Atwood,
H., 2004. Practical Diets for Litopenaeus vannamei (Boone. 1931): Working
Towards Organic and/or All Plant Production Diets. Avances en Nutricion Acuicola
VII. Memorias del VII Simposium Internacional de Nutricion Acuicola. 16-19
Noviembre, 2004. Hermosillo, Sonora, Mexico
Decamp, O., Cody, J., Conquest, L., Delanoy, G., Tacon, A.G.J., 2003. Effect of salinity on
natural community and production of Litopenaeus vannamei (Boone), within
experimental zero-water exchange culture systems. Aquac. Res. 34, 345-355.
Dersjant-Li, Y., 2002. The use of soy protein in aquafeeds. Avances en Nutricion Acuicola
VI. Memorias del VI Simposium Internacional de Nutricion Acuicola. 3 al 6 de
Septiembre del 2002. Cancun, Quintana Roo, Mexico.
Divakaran, S., Velasco, M., Beyer, E., Forster, I., Tacon, A.G.J., 2000. Soybean meal
apparent digestibility for Litopenaeus vannamei, including a critique of
methodology. Avances en Nutricion Acuicola V. Memorias del V Simposium
Internacional de Nutricion Acuicola. 19-22 Noviembre, 2000. Merida, Yucatan,
Mexico.
Food and Agriculture Organization FAO, 2005. FAO Fisheries Department, Fishery
Information, Data and Statistics Unit. FISHSTAT Plus: Universal software for
dtatistical time series. Version 2.3, 2000.
Forster, I.P., Dominy, W., Tacon, A.G.J., 2002. The use of concentrates and other soy
products in shrimp feeds. Avances en Nutricion Acuicola VI. Memorias del VI
Simposium Internacional de Nutricion Acuicola. 3-6 de Septiembre del 2002.
Cancun, Quintana Roo, Mexico.
Fox. J.M., Lawrence, A.L., Smith, F., 2004. Development of a low-fish meal feed
formulation for commercial production of Litopenaeus vannamei. Avances en
Nutricion Acuicola VII. Memorias del VII Simposium Internacional de Nutricion
Acuicola. 16-19 Noviembre, 2004. Hermosillo, Sonora, Mexico
Garza, A., Rouse, D.B., Davis, D.A., 2004. Influence of nursery period on the growth and
survival of Litopenaeus vannamei under pond production conditions. J. World
Aquac. Soc. 35 (3), 357-365.
Gomez-Jimenez, S., Gonzalez-Felix, M.L., Perez-Velazquez, M., Trujillo-Villalba, D.A.,
Esquerra-Brauer, I.R., Barraza-Guardado, R., 2005. Effect of dietary protein level
on growth, survival and ammonia efflux rate of Litopenaeus vannamei (Boone),
raised in a zero water exchange culture system. Aquac. Res. 36, 834-840.
66
Gonzalez-Rodriguez, B., Abdo de la Parra, I., 2004 Replacement of fish meal with co-
extruded wet tuna viscera and corn meal in diets for white shrimp (Litopenaeus
vannamei, Boone). Aquac. Res. 35, 1153-1157.
Hardy, R.W., 1999. Alternate protein sources. Feed Management 50, 25-28.
Hernandez, C., Sarmiento-Pardo, J., Gonzalez-Rodriguez, B., Abdo de la Parra, I., 2004.
Replacement of fish meal with co-extruded wet tuna viscera and corn meal in diets
for white shrimp (Litopenaeus vannamei, Boone) Aquac. Res. 36, 834-840.
Hertrampf, J.W., Piedad-Pascual, F., 2000. Handbook on Ingredients for Aquaculture
Feeds. Kluwer Academic Publishers (ed.). Dordrecht, The Netherlands. 573 p.
Josupeit, H., 2004. An overview of the world shrimp market. World Shrimp Markets 2004,
26-27 October 2004, Madrid, Spain. http://www.globefish.org/
Leber, K.M., Pruder, G.D., 1988. Using experimental microcosms in shrimp research: the
growth-enhancing effect of shrimp pond water. J. World Aquac. Soc. 19, 197-203.
Lim, C., Dominy, W., 1990. Evaluation of soybean meal as a replacement for marine
animal protein in diets for shrimp (Penaeus vannamei). Aquaculture 87, 53-63.
Lim, C., Ako, H., Brown, C.L., Hahn, K. 1997. Growth response and fatty acid composition
of juvenile Penaeus vannamei fed different sources of dietary lipid. Aquaculture
151, 143-153.
Lim, C., Klesius, P.H., Dominy, W., 1998. Soyabean products. International Aqua Feeds 3,
17-23.
Mendoza, R., De Dios, A., Vazquez, C., Cruz, E., Ricque, D., Aguilera, C., Montemayor,
J., 2001. Fishmeal replacement with feather-enzymatic hydrolyzates co-extruded
with soya-bean meal in practical diets for the Pacific white shrimp (Litopenaeus
vannamei). Aquac. Nutr. 7, 143-151
Moriarty, D.J.W., 1997. The role of microorganisms in aquaculture ponds. Aquaculture
151, 333-349.
Moss, S.M., Divakaran, S., Kim, B.G., 2001. Stimulating effects of pond water on digestive
enzyme activity in the Pacific white shrimp, Litopenaeus vannamei (Boone). Aquac.
Res. 32, 125-131
67
Moss, S.M., Pruder, G.D., 1995. Characterization of organic particles associated with rapid
growth in juvenile white shrimp, Penaeus vannamei (Boone), reared under intensive
culture conditions. J. Exp. Mar. Biol. Ecol. 187, 175-191.
NRC, National Research Council. 1993. Nutrient requirements of fish.
Piedad-Pascual, F., Cruz, E.M., Sumalangcay, A., 1990. Supplemental feeding on Penaeus
monodon juveniles with diets containing various levels of defatted soybean meal.
Aquaculture, 89: 183-191.
Samocha, T., Davis, D.A., Saoud, I.P., DeBault, K., 2004. Substitution of fish meal by co-
extruded soybean poultry by-product meal in practical diets for the Pacific white
shrimp, Litopenaeus vannamei. Aquaculture 231, 197-203.
Schuur A.M., 2003. Evaluation of biosecurity applications for intensive shrimp farming.
Aquac. Eng. 28, 3-19.
Steel, R.G.D., Torrie, J.H., 1980. Principles and Procedures of Statistics: A Biometrics
Approach. McGraw-Hill, New York, NY, USA.
Swick, R.A., Akiyama, D.M., Boonyaratpalin, M., Creswell, D.C., 1995. Use of soybean
meal and synthetic methionine in shrimp feed. American Soybean Association,
Technical Bulletin (Vol. AQ43-1995).
Tacon A.G.J., 1996. Nutritional studies in crustaceans and the problems of applying
research findings to practical farming systems. Aquac. Nutr. 1, 165-174.
Tacon, A.G.J., Barg, U.C., 1998. Major challenges to feed development for marine and
diadromous finfish and crustacean species. In: De Silva, S.S. (Ed.), Tropical
Mariculture. Academic Press, San Diego, CA, USA, 171 - 208.
Tacon, A.G.J., Cody, J.J., Conquest, L.D., Divakaran, S., Forster, I.P., Decamp, O.E., 2002.
Effect of culture system on the nutrition and growth performance of Pacific white
shrimp Litopenaeus vannamei (Boone), fed different diets. Aquac. Nutr. 8, 121-137.
Venero, J., 2006. Optimization of dietary nutrient inputs for Pacific white shrimp
Litopenaeus vannamei. Doctoral dissertation. Auburn University, Auburn,
Alabama, USA. .
Zelaya, O., 2005. An evaluation of nursery techniques and feed management during culture
of marine shrimp Litopenaeus vannamei. Doctoral dissertation. Auburn University,
Auburn, Alabama, USA.