Sample proposal on blue crabs

Diet development for blue swimmer crab, Portunus pelagicus juveniles, with
emphasis on lipids nutirition
PhD Candidate:
Supervisor:

1.0 Background, Aims and Significance of the Project
The blue swimmer crab, Portunus pelagicus also known as sand crab, is important in commercial
and recreational fisheries in Australia (Lestang et al., 2003). Catches for this species have risen
substantially from 200 tonnes in 1987/88 to 740 tonnes and valued $2.2 million in 1997/98 (Kangas,
2000). Increases in the catches of commercial fishers and the high participation of recreational
fishers (Williams, 1982; Kangas, 2000) reflect the value of this species to consumers and increasing
market demands. Hatchery culture of this species is relatively successful with good survival rates
(Josileen and Menon, 2005; Romano and Zeng, 2006), providing a solid basis for the fast expansion
of a farming industry for this species. However, although blue swimmer crabs farming is promising,
so far published information on culturing this species intensively is very limited. As the emerging of
aquaculture industry of the species puts a strong demands on feed used to grow them (Watanabe,
2002), a comprehensive and quantitative understanding of the nutritional requirements of the species
are critical to increase production and to facilitate further expansion of the industry.

One of the major concerns in crustacean nutrition is lipids nutrition. There is ample evidence from
previous studies showing that dietary lipids imbalance severely reduced growth rates, molting
frequency and survivals. Much previous work has been done on crustacean lipids requirement was
done on Penaied prawns, and so far there is so far no published information available on lipids
nutrition for blue swimmer crabs. There is clear evidence that lipids requirements of crustaceans are
species-specific. Therefore, systematic and quantitative studies on lipid requirements for the blue
swimmer crabs are necessary.

Lipids consist of varying classes and constituents, which includes phospholipids, fatty acids, sterols
and caratenoids (L.Gonzalez-Felix et al., 2002). Each of these components needs to be supplied in
1

optimum amount and balanced with other constituents. Phospholipids, normally supplemented in the
form of lecithin in formulated feed, and cholesterol, are bio-membranes components that are
important in maintaining cellular functions and structure (Gurr and Harwood, 1991). It is believed
that lecithin facilitates transport of cholesterol and has significant interactions when supplied
together in the diets (Gong et al., 2000). However, few studies have focused on their interactions. As
a cell component and a metabolic precursor of steroid, brain, vitamin D and molting hormones
(New, 1976; Gong et al., 2000; Hernandez et al., 2004), dietary cholesterol is also essential for the
growth of crustaceans.

Sargent (1999) pointed out that, essentiality of highly unsaturated fatty acids (HUFA),
ecosapentaenoic fatty acids, (EPA; 20:5n-3), docosahexaenoic fatty acids (DHA; 22:6n-3) and
arachidonic acids (ARA; 20:4n-6), and competitive interactions between them, are fundamentals to
lipids nutrition study. Crustaceans need supplemental dietary HUFA because of their inability to
elongate and desaturate polyunsaturated fatty acids (PUFA) to HUFA (L.Gonzalez-Felix et al.,
2002). Much research has been done on n-3 HUFA requirements in crustacean; however, less
information is available on the important of optimal ratio of EPA to DHA. Because both DHA and
EPA play major roles in survival and growth of crustaceans, it is possible that n-3 HUFA
requirements are not only a function of total amount but also the proportions of EPA and DHA.

Furthermore, less attention of the past dietary HUFA investigation in crustaceans has been given to
arachidonic acid as essential fatty acids (EFA). Eicosanoids that are produced from ARA are more
biologically active than those produced from EPA (Sargent et al., 1999) and it plays important roles
in crustaceans molting (Sheen and Wu, 1999). Although it has been reported, removing ARA from
P. monodon diets resulted in no notable response in growth rate (Merican and Shim, 1996),
conservation of ARA during starvation signifies some important functions of ARA (Wen et al.,
2006). Lack of research on ARA requirements in crustaceans is believed a major attributor for the
uncertainty of nutritional values of ARA as a major HUFA for crustaceans and further investigation
is needed.

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Nutrition is one of the core factors in blue swimmer crab cultivation and needs to be explored in
great details for further development of this species. A good formulated diet not only will boost the
productions of cultured species, but will also help maintaining water quality. Particle break downs
from diets has been suggested to cause deterioration of water quality (Meyers et al., 1972).
However, water stability of the diets can be increased by using binders (New, 1976) that helps form
compact and durable pellet (Silva and Anderson). Still, binders may also impair digestibility of feed
and loss of nutritional content due to high leaching rate of water soluble nutrients (Meyers et al.,
1972; New, 1976; Genodepa, 2004). Although some nutrients leaching may be necessary to
stimulate feed intake (Baskerville-Bridges and Kling, 2000), the balance between leaching rate and
amount of nutrients left for culture animals to consume is far more important.

Nutrient levels and water stability are not the only important factor in developing formulated diets.
The quantity of food consumed also has pronounced effects on growth rate, efficiency of food
conversion and chemical composition (Reinitz, 1983) of culture organisms. In addition, this factor is
also water correlated with the size of culture organism and water temperature (Khan et al., 2004).
The significant difference of food ration and temperature is mostly important in region where there
are noticeable changes in temperatures (e.g. winter and summer) where the amount of food can be
adjusted to suit culture organism requirements. As optimization of feeding rate of culture animals
are important to achieve efficient production (Khan et al., 2004), the final stage of this study will
focus on finding the right ration for blue swimmer crabs at different water temperatures.

In summary, the main aims of the proposed PhD project are to quantify the optimum amount of
dietary lipids and its constituents that are needed for the early blue swimmer crab juveniles. The
research outcomes will develop formulated diets for blue swimmer crabs and will have good
potential of being commercialized. A quantitative and qualitative approach will be used to achieve
the following objectives:
1. To investigate variation on lipids composition of the early P. pelagicus juveniles under normal

feeding and various starvation condition;
2. To determine total dietary lipids requirements of P. pelagicus juveniles;

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3. To evaluate potential interactions of lecithin and cholesterol at different dietary levels;
4. To determine n-3 HUFA requirements of P. pelagicus juveniles;
5. To evaluate potential interactions of DHA (22:6n-3) and EPA (20:5n-3) at different dietary
level;
6. To determine n-6 HUFA, arachidonic acid requirements of P. pelagicus juveniles;
7. To optimize binder component for diets fed to P. pelagicus juveniles; and
8. To investigate effects of water temperature and ration on survival and growth of P. pelagicus
juveniles, in individual and communal culture.

2.0 Research Plan
2.1

2.1.1

General Materials and Method

Source of experimental crabs

Juvenile blue swimmer crabs to be used in all experiments will be reared in laboratory. Wild caught
broodstocks will be maintained in 1000 L outdoors tanks until spawning. Berried females will be
transferred indoor and kept in 300 L tank. Upon hatching, larvae will be cultured with rotifers and
Artemia until settlement as fist stage crabs (C1) based on protocols established at JCU (Romano and
Zeng, 2006).

2.1.2

Experimental setups

As cannibalism could substantially affect the results experiments, experiments 1 to 7 will be
conducted by housing individual crabs in 750 ml perforated plastic containers (7.5 cm diameter and
12 cm height). The containers will be labeled for treatments and will be randomly distributed in
water baths for temperature control.

The experiments are expected to be carried out mainly from crab stage one to five (C1 to C5).
Except for Experiment 1 and 8, a minimum number of 20 individually kept crabs will be used for
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each treatment and the treatment will be triplicated. Experimental crabs will be fed every morning
until satiation and any food leftover will be siphoned out before each feeding. Mortality and molting
will be recorded daily.

100% of water changes will be conducted every day. Once per week, DO, pH and ammonia level
will be checked to ensure good water quality. Temperature, salinity and pH will be maintained at
28oC, 30 ppt and 8.5, respectively. Throughout the experiments, photoperiod will be set at 12h ratio
(light:dark).

2.2

Diet Preparation

Except for experiment 1, formulated food will be prepared and the ingredient of the basal diet as
listed in Table 1 will be used except some necessary modifications in starch and cellulose levels to
maintain isocaloricity of the diets used in the same experiment. Before preparations of the diets,

lipids will be extracted from basal ingredients using hot ethanol to minimize the contribution of
other dietary lipids (Sheen 2000). The feeds will be crumbled and sieved to get desired size
(< 2mm). It will be stored at -20oC to prevent rapid oxidation of lipids and remain the quality of the
diets. To obtain persist and reliable results in growth trial, prior to experiments commence, the diets
will be tested for their stability in water and crabs acceptability.
Table 1: Basal mix of diets (modified from Sheen and Wu 1999; 2000)
Ingredient
Defatted squid meal
Vitamin mixturea
Mineral Mixtureb
Choline chloridec
DCPd
Zein e

Source
Skretting Tasmania
Rabar Pty Ltd
Rabar Pty Ltd
Sigma-Aldrich Pty Ltd
Sigma-Aldrich Pty Ltd
Sigma-Aldrich Pty Ltd
Total

a)

% of dry weight
50%
4
4
1

0.6
3
62.6%

ZZ603 DO 067 DPI, each 1kg contains: copper 1g, cobalt 100mg, magnesium 59.4mg, manganese
5g. iodine 800mg, selenium 20mg, iron 8mg, zinc 20g, aluminium 100mg, chromium 100mg b)
ZZ600 DPI, each 1kg contains: vitamin A 2miu, vitamin D3 0.8miu, vitamin E 40g, vitamin K
2.02g, inositol 50g, vitamin B3 30.40g, vitamin B5 9.18g, vitamin B9 2.56g, vitamin B2 4.48g,
vitamin B12 0.004g, biotin 0.1g, vitamin B6 4g, vitamin B1 3.4g, vitamin C 44.4g, para amino

5


benzoic acid 20g, tixosil 5g, antioxidant 30g c) 98% powder C7527 d) dibasic calcium phosphate
C4131 e) Z3625 from maize

2.3

Proximate and Fatty Acid Analysis of Diets and Crabs

The diets and sample of pooled whole crabs at the beginning and termination of the experiment can
be analyzed for proximate compositions and fatty acids based on the AOAC (1984) methods (Sheen
and Wu, 2003). Crude protein will be determined using Kjedahl method while total lipid content in
the samples will be determined by the chloroform-methanol (2:1, v/v) extraction method (Folch et
al. 1957). Ash and moisture can be measured using muffle furnace and laboratory oven. Fatty acids
analysis can be done based on modified method reported by Sheen and Wu (1999).

2.4

Data Collection

For all experiment carapace length and width, wet weight and survival of crabs will be recorded for
each molt. Samples will also be taken for histology and at the end of experiments for dry weight
measurement. Depending on experimental design, either one-way or two-way ANOVA will be used
for data analysis with Tukey’s test to identify significant differences among treatments.

Experiment 1: Effects of starvation on growth, survival and tissue biochemical and fatty acid
profiles of blue swimmer crabs early juveniles
Rationale
Effects of starvation on utilization of nutrients have been studied in crustaceans to gain more
understandings on their requirements (Barclay et al., 1983; Clifford and Brick, 1983; Dall and
Smith, 1986; Wen et al., 2006). A study conducted on juvenile Chinese mitten crab showed that
PUFA and HUFA were conserved in both hepatopancreas and muscle of the crab at the expense of
saturated and monounsaturated fatty acids (Wen et al., 2006). It has been proved that conservation of
PUFA and HUFA are because of their importance as cell structure. Outcomes of this experiment will
help understanding the preferential of lipid constituent conserved during starvation for blue
swimmer crabs and thus provide useful clues on lipids requirements of this species.

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Methodology
A total of 656 juvenile crabs will be used and will be divided to 3 treatments, i.e fed (F), starved (S)
and starvation for 7 days and re-fed (7 S-F). Crabs will be fed with commercial pellet produced by
Ridley® for the tiger shrimp, Penaeus monodon, which contains 43% protein, 6% fat and 3% fiber.
Samples will be taken 3 times for all treatments during the experiment for proximate and fatty acids
analysis as well as histology.
Table 2: Time when sample will be taken
Treatment
3rd day

Fed (F)
Starved (S)
7 Day Starvation and Re-fed (7 S-F)

7th day

▪
▪

▪
▪

10th day

C2

C3

▪

▪

▪

▪

▪

Expected Outcomes
•

Data on survival, development and growth as well proximate and fatty acid profile and
histochemistry of digestive gland will be used to assess preferentially conserved and utilized
of major nutrients and lipids in both fed and those under various starvation conditions.

Experiment 2: Effects of various dietary lipid levels on the survival, development and growth
of the blue swimmer crabs juvenile
Rationale
Levels of lipids in the diets are closely associated with crustaceans survival, development and
growth, thus there are substantial interest in determining the optimal total lipids that a particular
crustacean needs (Sheen and Wu, 1999). For crustaceans, both inadequate and excessive lipid levels
negatively impact their grow-out. Low level of dietary lipids may result in low molting frequency
(Sheen and Wu, 1999) and at high levels, retarded growth (Ackefors et al., 1992). Requirements of
total lipids for crustaceans seem to be species-specific, although generally it is within 5-10% range
(Sheen and Wu, 1999).

Methodology
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Seven diets with lipid levels ranging from 0-12% at 2% increments and 2:1 ratio (fish oil:corn oil)
(Sheen and Wu, 1999) will be formulated and used to feed crab juveniles. For the production of
crabs for the experiment and other experimental design, data collection, please refer to ‘General
materials and method’ (subsection 2.1).
Expected outcomes
•

Established optimal total lipids levels in the diets for the blue swimmer crabs juveniles

Experiment 3: Determining optimum levels and ratios of dietary cholesterol and lecithin and

their effects on survival, growth performance and tissue lipid composition for
blue swimmer crab juveniles
Rationale
Lecithin play an important role in lipid mobilization and is believed that it facilitates transport of
cholesterol (Gong et al., 2000). Cholesterol is a compound of sterols and important as a cell
component and is a metabolic precursor of brain, vitamin D and molting hormones (New, 1976;
Gong et al., 2000; Hernandez et al., 2004). It has been proven that imbalanced cholesterol and
lecithin in the diets negatively impact crustacean growth (Chen and Jenn, 1991). This highlights the
need to investigate balanced supply of cholesterol and lecithin in the blue swimmer crab diets.
Methodology
Three level of cholesterol of 0%, 0.5% and 1.0% will be paired with three levels of lecithin, 2%, 4%
and 6%. Levels of both cholesterol and lecithin were chosen based on literature on range required
for crustaceans. All nine diets will be evaluated based on survival and growth performance of blue
swimmer crabs juvenile. Effects of levels of dietary cholesterol and lecithin on tissue fatty acids
composition will also be investigated.
Expected outcomes
•

The optimal level of lecithin and cholesterol for the blue swimmer crab juveniles will be
determined and used as the basis for formulating diets in following experiment.

•

Understanding the potential compensatory effects of lecithin and cholesterol.

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Experiment 4: Determining the optimal levels of n-3 HUFA for blue swimmer crab juveniles
Rationale

n-3 highly unsaturated fatty acids (n-3 HUFA) that contains eicosapentaenoic acid (EPA, 20:5n-3)
and docosahexaenoic acid (DHA, 22:6n-3), plays important physiological roles as components of
membrane phospholipids and as precursors of biologically active eicosanoids (Bell et al., 1986;
Mourente and Tocher, 1992). Due to their various function and inability of crustaceans to synthesize
their own n-3 HUFA, inclusion of n-3 HUFA in most favorable amount is essential to boost good
survival and growth. Although, the importance of n-3 HUFA to crustaceans species are well
documented, the requirements of n-3 HUFA for blue swimmer crab is unknown.
Methodology
For this experiment, six diets containing levels of n-3 HUFA from 0 to 2.5% with a 0.5% increment
will be formulated. Inclusion of cholesterol and lecithin in all diets will be based on best interaction
levels of these compounds in previous experiments. Fish oil will be used as source of n-3 HUFA and
will be adjusted to desired n-3 HUFA levels that will be tested in this experiment.
Expected Outcomes
•

The optimal level of n-3 HUFA in the diets of blue swimmer crabs juvenile will be
determined.

Experiment 5: Identifying optimal ratio of EPA (20:5n-3) and DHA (22:6n-2) for blue
swimmer crab juveniles
Rationale
Among the numerous study on n-3 HUFA requirements for crustaceans, relative little attention have
been given to address requirements of EPA and DHA individually (Read, 1981; Merican and Shim,
1996). Of those relative few research, most of it often focused on DHA requirements (Merican and
Shim, 1997; Mourente and Rodriguez, 1997). Suprayudi (2004) suggested that DHA is more
efficacious than EPA in promoting growth. However, the fact that EPA is important in maintaining
survival and involved in producing eicosanoids (3-series prostanoids and 5-series leukotrienes)
(Sargent et al., 1999) indicating that both of the fatty acids are important and balanced proportions of

9

these compound might be crucial. Study on P. monodon illustrated adverse effects on weight gain
when fatty acids ratio was imbalanced (Glencross et al., 2002b). The study showed that best growth
was obtained when same amount of EPA and DHA were supplied.
Methodology
Total level of n-3 HUFA in this experiment will be based on the best result in n-3 HUFA experiment
(Exp 4). However, in formulating the diets, EPA and DHA will be adjusted to match the tested ratio.
Source of EPA and DHA will be from purified blended n-3 HUFA fish oil. All seven diets will be
fed to blue swimmer crab juveniles following protocols described in General Materials and Methods
(subsection 2.1).
Table 3: Percentage of EPA and DHA and their ratio in tested diets
EPA (%)

0

100

50

35

65

25

75

DHA (%)

100

0

50

65

35

75

25

Ratio (EPA:DHA)

0:1

1:0

1:1

1:2

2:1

1:3

3:1

Expected Outcomes
•

Determine the optimum dietary EPA and DHA ratio for blue swimmer crab juveniles.

Experiment 6: Determining the optimal levels of arachidonic acid, ARA (20:4n-6) for blue
swimmer crab juveniles
Rationale
Molting process in crustaceans are regulated by eicosanoids substances derived from ARA
(L.Gonzalez-Felix et al., 2002). When Penaeus esculentus when injected with prostaglandin E2, a
type of eicosanoids, shorter molt cycles and better growth were displayed compared to control group
(Koskela et al., 1992). As a HUFA constituent, ARA also could not be internally synthesized by
crustaceans and need to be provided through the diets. Lack of n-6 fatty acids in the diet have been
proven to have negative effects on growth (Reigh and Stickney, 1989; Glencross and Smith, 1999).
Despite the important of ARA to crustaceans, only limited information on quantitative requirements
of ARA are available.

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Methodology
Six diets will be formulated containing ARA from 0 to 2% with 0.5% increments. All diets will be
formatted to contain optimum level of n-3 HUFA and best ratio of EPA and DHA based on
experiments 4 and 5.
Expected outcomes
•

Optimum level of ARA requirement for blue swimmer crab juveniles.

•

Understanding the effects of n-6 HUFA and n-3 HUFA on blue swimmer crab juveniles.

Experiment 7: Optimizing binder compound in formulated diets fed to blue swimmer crab
juveniles
Rationale
Various studies have assessed the use of various binders in formulated diets for aquaculture animals
(Knauer et al., 1993; Pearce et al., 2002; Genodepa, 2004; Ruscoe et al., 2005). Inclusion of binder
is necessity to ensure water stable pellets (Cuzon et al., 1994). Stability of the feeds in water is
especially important for crustaceans which generally find food by chemoreception rather than sight,
and may not consume it for several hours (D'Abramo, 1997). However, some binding agents were
claimed to impair the digestibility of animal feeds (New, 1976) and may also results in diets that are
not adequately stable and cause excessive nutrient loss and water pollution (Genodepa, 2004).
Methodology
Diets with optimum levels of lipids, based on previous experiment, will be formulated with 4
different types of binders; zein, gelatin, alginate and agar at the same level (3%) (Genodepa, 2004).
The diets will be evaluated by measuring physical parameters of the diets in seawater column (water
stability and leaching rate) and nutritional performance of the diets (survival and growth). Water
stability of the diets will be based on modification of method used by Ruscoe et al. (1995). Dry
matter remaining (DMR) of the diets after interval periods of immersion will be determined. DMR
will be determined using following equation:
DMR (%) = Wo x (1-M) –Wt x100
Wo X (1-M)

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Wo = initial diet weighed, Wt = diet weight after immersion and drying, M = moisture content of diet
as a proportion.
Leaching rate of the diets will be evaluated using methods proposed by Baskerville-Bridges et al.,

(2000). Diets with different binders will be formulated separately by adding free amino acid,
glycine. Glycine via leaching after various intervals immersion will be calculated using ninhydrin
reagent and spectrophotometer. Diets without any addition of binder will be used as a control and
will be tested in the same way.
Table 4: Time when sample will be taken
Diet Immersion Period
Test
Water
stability
Leaching
rate

1 min

15
min

30
min

▪
▪

▪

60
min

2h

3h

6h

12h

18h

24h

▪

▪

▪

▪

▪
▪

▪

▪

▪

Diets with different binders will be fed to crabs based on feeding methods used in previous
experiments. Diets performance will be determined on the basis of survival and growth.
Expected outcomes

•

Water stability of the diets at different time immersion.

•

Leaching rate of diets with different binders.

•

Crabs acceptance toward the diets with different binders.

•

Best binder will be used to formulate diets for next experiment.

Experiment 8: Effects of water temperature and food ration on growth performance of blue
swimmer crab juveniles, in individual and communal culture.
Rationale
Optimization of feeding rate of culture animals is important to achieve efficient production (Khan et
al., 2004) and temperature is among important factors that controlled growth of crustaceans (Wyban
et al., 1995). Temperature directly effects the rate of physiological process (Spanopoulous-

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Hernandeza et al., 2005) and thus elevated metabolic rate of crustaceans at higher temperatures
(Allan et al., 2006). This is parallel with increased in feeding rates at increased water temperature
levels in P. vannamei juveniles (Wyban et al., 1995). Information on food ration and temperature
manipulation will be useful in culturing blue swimmer crabs at different time of the year.

Methodology
Diets in this experiment will be formulated based on previous results. It will be then fed to crabs at 4
different feeding rates, (5%, 10%, 15% and 20% of wet weight) and two different temperatures;
22oC and 30oC as in winter and summer condition. Crabs will be cultured individually and also
communally in twenty four 60 L perforated containers. The containers will contains 20 crabs each
and will be submerged in eight 300 L tanks with three containers placed randomly in each tank.
Crabs will be individually and communally culture to determine growth and survival rate between
two culture conditions. Metabolic rate of the crabs will be determined using a modified method used
by Allan (2006) to verify if there any changes in metabolic rate at tested temperatures. Crab will be
culture individually in 100 ml conical flask sealed with lid and petroleum jelly for 1 h at tested
temperature. Control flask will be prepared in the same way without any crab. The oxygen
concentration between control and tested flask and crab’s wet weight will be measured using DO
oxygen meter and electronic balance after 1 h. Mass specific oxygen consumption rates of the crabs
will be expressed as microliter of oxygen consumed per milligram of wet weight per hour (µl O2 mg
wet weight-1 h-1).
Expected Outcome
•

Best food ration for blue swimmer crabs at different water temperatures.

•

Growth and survival rate differences between individually culture and communally culture
crabs.

•

Metabolic rate changes at different temperature.

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3.0 Timeline

Tasks

06
A

S

O

07
N

D

J

F

M

A

M

08
J

J

A

S

O

N

Writing
Literature
Review
Proposal
Thesis
Research
Exp 1
Data Analysis
Exp 2
Data Analysis
Exp 3
Data Analysis
Exp 4
Exp 5
Exp 6
Data Analysis
Exp 7
Data Analysis
Exp 8

Data Analysis

14

D

J

F

M

A

M

09
J

J

A

S

O

N

D

J

F

M

A

M

J

J


4.0

Budget

1.

Broodstock food………………………………………………

$ 1,200

2.

Equipment…………………………………………………….

$ 1,000

3.

Chemical………………………………………………………

$ 200

4.

Diet ingredients……………………………………………….

$ 2,000

5.

Analysis……………………………………………………….

$ 6,400

Estimated Cost of Project……………………………………………..

$10,800

5.0
1.

Justification of Budget
Broodstock food: The price of mussel, squid and prawns need for feeding crabs for one month
is estimated at $ 100, for total 3 years of study, $ 3,600. The amount will be shared with two

other post graduate student working on crab species, $ 3,600/3 = $ 1 200

2.

Equipment: The amount is needed for plastic containers, mesh, heater for crabs culture and
vial, polysterene box for samples collecting and storage.

3.

Chemical: Antibiotics, chlorine, alcohol and formalin will be used mainly for sterilizing
laboratory and keeping the crabs from any parasite infections.

4.

Diet ingredients: High quality diets will be bought from Sigma-Aldrich Company Pty Ltd,
Rabat Pty Ltd, Skretting Tasmania and health store.

5.

Analysis: One analysis cost $ 100, a mean of 8 analysis per experiment will be needed for each
8 experiment, $ 100*8*8 = $ 6 400

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6.0

Publications Plan

Based on experiments that will be done in this study, 8 potential publications can be developed and

published during 3 years of study:
1. Nutritional requirements and starvation resistance in juvenile blue swimmer crabs, Portunus
pelagicus
2. Effects of dietary lipid on growth performance and body lipid composition of blue swimmer
crabs (Portunus pelagicus)
3. Combine effects of dietary lecithin and cholesterol on the growth, survival and body lipid of
marine crabs, Portunus pelagicus at juvenile stage
4. Essential fatty acids in the diet of blue swimmer crabs juvenile (Portunus pelagicus): effects of
n-3 HUFA
5. Essential fatty acids in the diet of blue swimmer crabs juvenile (Portunus pelagicus) : effects
of EPA/DHA ratio
6. Essential fatty acids in the diet of blue swimmer crabs juvenile (Portunus pelagicus) : effects
of n-6 HUFA, Arachidonic acid (20:4n-6)
7. Effects of binder on formulated diets fed to blue swimmer crabs Portunus pelagicus
8. The effects of water temperature and food ration on growth and survival of blue swimmer
crabs juvenile (Portunus pelagicus)
References:
Ackefors, H., J. D. Castell, et al. (1992). "Standard experimental diets for crustacean nutrition
research. II. Growth and survival of juvenile crayfish Astacus astacus fed diets containing various
amounts of protein, carbohydrate and lipid." Aquaculture 104: 341-356.
Allan, E. L., P. W. Froneman, et al. (2006). "Effects of temperature and salinity on the standard
metabolic rate (SMR) of the caridean shrimp Palaemon peringueyi." Journal of Experimental
Marine Biology and Ecology 337: 103-108.

16


Barclay, M. C., W. Dall, et al. (1983). "Changes in lipid and protein during starvation and the
moulting cycle in the tiger prawn, Penaeus esculentus Haswell." Journal of Experimental Marine
Biology and Ecology 68(3): 229-244.

Baskerville-Bridges, B. and L. J. Kling (2000). "Development and evaluation of micropartculate
diets for early weaning of Atlantic cod Gadus mohua larvae." Aquaculture Nutrition 6: 171-182.
Bell, M. V., R. J. Henderson, et al. (1986). "The role of polyunsaturated fatty acids in fish."
Comparative Biochemistry and Physiology 4: 711-719.
Chen, H. Y. and J. S. Jenn (1991). "Combined effects of dietary phosphatidylcholine and cholesterol
on the growth, survival an body composition of marine shrimp Penaeus penicillatus." Aquaculture
96: 167-178.
Clifford, H. C. and R. W. Brick (1983). "Nutritional physiology of the freshwater shrimp
macrobrachium rosenbergii (de man)-I. Substrate metabolism in fasting juvenile shrimp."
Comparative Biochemistry and Physiology Part A: Physiology 74(3): 561-568.
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