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Department of Food Science, National Taiwan Ocean University Keelung, Taiwan 202 ROC
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ABSTRACT |
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 TOP ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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KEY WORDS: • pantothenic acid • shrimp • Penaeus monodon
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INTRODUCTION |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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, Soliman and Wilson 1992
), channel catfish (Wilson et al. 1983
),
common carp (Ogino 1967
) and salmon (National Research Council 1993
), respectively.
For crustaceans, only limited qualitative data concerning vitamin
requirements are available. Kanazawa (1985)
reported the
requirements for larval Penaeus japonicus of various
vitamins, including PA. Liu et al. (1995)
also reported
that Penaeus chinensis required dietary PA for optimum
growth. However, quantitative information on the requirements of PA for
penaeid shrimp is lacking. To confirm estimates of aquatic species' of
dietary requirement water-soluble nutrients, in general, the
quantification of nutrient is important. The purpose of this study was
to define the adequate dietary PA requirement for juvenile grass
shrimp, Penaeus monodon, using growth indices supported by
measurement of tissue PA and coenzyme A concentrations.
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MATERIALS AND METHODS |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Experimental diet formulation is given in Table 1.
The formulation is similar to that of Shiau and Hwang (1994)
, which has been shown to be adequate for P.
monodon. Vitamin-free casein (Sigma Chemical, St. Louis, MO), fish
oil (Scott and Bowne, London, U.K.), and corn starch (Sigma Chemical)
were used as dietary protein, lipid, and carbohydrate sources,
respectively. A mixture of amino acids [including glycine, L-alanine,
L-glutamate, and betaine (Sigma Chemical)] was included in the diets
as an attractant (Shiau and Jan 1992
). The vitamin
mixture was similar to that used by Shiau and Yu (1998)
,
except that it did not contain PA. Pantothenic acid (Sigma Chemical)
was added to the test diets, at the expense of small amounts of
cellulose, to provide concentrations of 0, 20, 40, 60, 120, 240, and
480 mg PA/kg diet. The PA concentrations of the seven diets were
determined by high pressure liquid chromatograph (Wyse et al. 1985
) to be 0.02 (unsupplemented control), 17.3 (20 mg/kg),
34.9 (40 mg/kg), 51.7 (60 mg/kg), 102.6 (120 mg/kg), 209.8 (240 mg/kg),
and 419.7 (480 mg/kg). The diets were prepared and stored as previously
described (Shiau and Yu 1998
).
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Juvenile P. monodon were supplied by the Tungkang Marine Laboratory (Tangkang, Pingtung, Taiwan). Upon arrival, they were acclimated to laboratory conditions for 2 wk in a plastic tank [74 cm (w) x 95 cm (1) x 45 cm (h)] and fed a commercial diet (grass shrimp no. 2 feed, Yung-Hsien, Taipei, Taiwan). The laboratory conditions during the acclimation period were similar to those at the initiation of the experiment. The proximate composition (g/100 g) of the commercial diet was as follows: moisture, 9.36; crude protein (N x 6.25), 37.30; lipid, 5.03; and ash, 12.53. At the beginning of the experiment, 21 aquariums (60 x 60 x 45 cm3) were each stocked with 22 shrimp with an average weight of 0.88 ± 0.01 g. Each experimental diet was fed to three groups of shrimp. Each aquarium received continuous aeration. In each aquarium, impurities from uneaten feed and fecal pellets in the water were removed by siphoning every day, and 75% of the water was exchanged at 2, 4, and 6 wk to maintain water quality. Dissolved oxygen concentration was monitored weekly and maintained at 7.5 mg O2/L throughout the experimental period. Water temperature ranged from 25 to 29°C, pH from 6.3 to 6.5, and salinity from 19 to 21 g/kg. A photoperiod of 12 h light, 12 h dark (0800–2000 h) was used. Shrimp were fed their respective diets at a rate of 8 g/100 g body weight. This daily ration was subdivided into two equal feedings given at 0830 and 1730 h. Shrimp were weighed biweekly, and the daily ration was adjusted accordingly. Any dead shrimp were not replaced during the experiment. The duration of the study was 8 wk.
At the end of the feeding trial, the shrimp were weighed, and three
were selected randomly from each aquarium and killed. Hepatopancreata
were quickly removed and pooled. Coenzyme A (CoA) and its thiol-esters
were extracted from the tissues (Lopaschuk et al. 1986
)
and measured according to Michal and Bergmeyer (1985)
.
The remaining hepatopancreata were used for PA (Wyse et al. 1985
) and lipid (Folch et al. 1957
)
determinations. Growth (as measured by the percentage of body weight
gain), feed efficiency (FE) and the protein efficiency ratio (PER) were
calculated as described previously (Shiau and Chou 1991
,
Shiau and Liu 1994
).
Statistical analysis.
Each experimental diet was fed to three groups of shrimp. Growth data
were means of three groups of shrimp, with 22 shrimp per group
(n = 3). Hepatopancreatic PA, CoA, and lipid
concentrations were means of the three groups of shrimp, with three
shrimp randomly selected from each group and pooled (n = 3). Results were analyzed by one-way ANOVA. When the ANOVA identified
differences among groups, multiple comparisons among means were made
with Duncan's new multiple range test. Statistical significance was
determined by setting the aggregate type I error at 5%
(P < 0.05) for each set of comparisons. Dietary PA
requirements for juvenile P. monodon were estimated by the
broken-line method (Robbins 1986
).
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RESULTS |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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).Patterns of difference in PER were similar to those of the FE ratio.
The survival percentage was significantly higher in shrimp fed diets
supplemented with PA than in shrimp fed the unsupplemented, control
diet.
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60 mg PA/kg. The
differences among the three groups were significant.
Broken-line analysis was used for estimating the adequate requirements
of dietary PA (as shown in Fig. 1
).Based on the weight-gain percentage, the regression equations were
Y = 0.71X + 56.84, Y = -0.03X + 159.6. The
equations for hepatopancreatic CoA were Y = 0.72X + 140.70,
Y = 0.06X + 214.7. And the equations for PA were Y =
0.3575X + 19.65, Y = 0.00135X + 54.55. Because the
breakpoints at 138.86 ± 6.53 (weight gain), 112.14 ± 5.11
(CoA), and 101.42 ± 5.24 (PA) gave the least mean square error,
the adequate amount of dietary PA for juvenile P. monodon
was estimated to be 101-139 mg/kg diet.
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DISCUSSION |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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). In the present study, mortality
was exceptionally high in shrimp fed the PA-deficient diet compared to
shrimp fed the PA-supplemented diets.
Coenzyme A and phosphopantetheine are the recognized conenzyme
forms of the water-soluble vitamin PA. They are involved in >100
different reactions in intermediary metabolism, including some of the
most fundamental in carbohydrate, amino acid, and lipid metabolism
(Robisshaw and Neely 1985
). As the universal carriers of
acyl groups, they are particularly important in fatty acid metabolism.
In fatty acid synthesis and degradation, pantothenate coenzymes carry
the acids as acyl groups through repetitive synthetic or degradative
cycles. Fatty acids must also be activated by CoA before they can be
synthesized into triglycerides. This may explain higher the
hepatopancreatic lipid concentration in shrimp fed diets with lower
levels of PA (
40 mg/kg, Table 2
).
Weight gain is often used for estimating nutrient requirements in
animals. The altered CoA levels found in the present study suggest that
this variable may permit a satisfactory evaluation of PA status of the
shrimp. The broken-line analyses of weight gain, hepatopancreatic CoA
concentration, and PA concentration in P. monodon (Fig. 1)
suggest that hepatopancreatic CoA concentration and PA concentration
can be used to estimate the PA requirements of shrimp.
The PA requirement for growth was reported in a few aquatic animals.
For fish, a requirement of 10 mg/kg, 30 mg/kg, 30-50 mg/kg, and 40-50
mg/kg were reported for tilapia (Soliman and Wilson 1992
, Roem et al. 1991
), channel catfish
(Wilson et al. 1983
), common carp (Orgino
1967
), and salmon (NRC 1993
), respectively.
There was only one study in the literature that relates to crustaceans.
Liu et al. (1995)
reported that 100 mg PA/kg is
sufficient for Penaeus chinensis. However, no formal
estimates of dietary requirements were provided in that study because
only two dietary PA levels (50 and 100 mg/kg) were employed in that
study. Our results indicates that the PA requirement for P.
monodon is higher than that for fish. Aquatic crustaceans, unlike
fish, are slow feeders, and food particles usually remain suspended in
the water for extended periods before being consumed. Manipulation of
food particles by shrimp during feeding may further increase leaching
of nutrients. Therefore, a higher PA requirement for crustaceans than
for fish seems reasonable.
Leaching is always a concern in a nutrition study of aquatic species.
The experimental diets used in the present study were relatively stable
in terms of the stability of water-soluble nutrients because leaching
from a similar diet was minimal after 2 min in water (Bordner et al. 1986
). Our experiment was conducted under natural
conditions, thus the value obtained in the present study is relevant.
Such higher requirements for P. monodon than for fish, for
water soluble vitamins such as thiamin (Chen et al. 1991
), riboflavin (Chen and Hwang 1992
), biotin
(Shiau and Chin 1999
), and ascorbic acid (Shiau and Jan 1992
), were also demonstrated.
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FOOTNOTES |
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1 Supported by a grant from the National Science
Council of the Republic of China, grant number NSC 87-2313-B-019-012.
2 The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 USC section 1734 solely to indicate this fact.
4 Abbreviation used: CoA, coenzyme A; FE, feed
efficiency; PA, pantothenic acid; PER, protein efficiency ratio.
Manuscript received September 16, 1998. Initial review completed October 28, 1998. Revision accepted December 1, 1998.
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REFERENCES |
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TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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