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Growth, survival and macronutrient composition of Penaeus monodon Fabricius larvae fed with Chaetoceros calcitrans and Tetraselmis chuii
Aquaculture, 29 (1982) 253-260 Elsevier Scientific Publishing Company,
253 Amsterdam
- Printed
in The Netherlands
GROWTH, SURVIVAL AND MACRONUTRIENT COMPOSITION OF PENAEUS MONODON FABRICIUS LARVAE FED WITH CHAETOCEROS CALCITRANS AND TETRASELMIS CHUII
EMILIA
TOBIAS-QUINITIO
Aquaculture Department, Iloilo (The Philippines) (Accepted
12 December
and CESAR
T. VILLEGAS
Southeast Asian Fisheries Development
Center, P.O. Box 256,
1981)
ABSTRACT Tobias-Quinitio, E. and Villegas, C.T., 1982. Growth, survival and macronutrient composition of Penaeus monodon Fabricius larvae fed with Chaetoceros calcitrans and Tetraselmis chuii. Aquaculture, 29: 253-260. Penaeus monodon larvae were reared from zoea, (Z,) to mysis, (M,) using two different algal feeds, Chaetoceros calcitrans and Tetraselmis chuii. Artemia nauplii were added to both treatments at mysis, . Mean survival and growth rates in both treatments were different at 5% level of significance on the second and third day of culture, but did not differ during the mysis stage and the end of the g-day culture period. Z, and M, larvae fed with C. calcitrans had a lower crude protein but a higher lipid content than T. chuii-fed larvae. Differences in carbohydrate content were noticed in M, larvae. The implication of the findings are discussed. INTRODUCTION
Various species of phytoplankton have been used as food in the culture of penaeid larvae (Simon, 1978). Griffith et al. (1973) showed that the growth of P. setiferus and P. aztecus larvae was more rapid with Tetraselmis than Skeletonema. The use of the diatom, Chaetoceros grucilis as exclusive food for P. styEirostris and P. uannamei zoea (Z) to mysis (M) was effective in obtaining a survival rate of 84.8% (Simon, 1978). Other genera of phytoplankton such as Isochrysis, Cylindrotheca (Aquacop, 1977), Phaeodactylum (Heinen, 1976) and Nitzschia (Hirata et al., 1975) have been used in penaeid culture. Some increased survival and growth rates of the larvae, while others gave good growth but poor survival, and vice versa. These differences may be due to size or chemical composition of algal cells. There is a continuous need to evaluate the effects of the algal food on the survival, growth and chemical composition of the prawn in order to establish good working criteria for the production of P. monodon postlarvae. This study investigates the effects of two species of phytoplankton, the diatom Chaetoceros calcitruns and the flagellate Tetruselmis chuii, on the growth and survival
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0 1982
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Scientific
Publishing
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of P. monodon; compares the nutritive values of C. calcitruns and T. chuii, and the effects of both on the chemical composition of whole larvae at the zoea and mysis stages. MATERIALS
AND METHODS
P. monodon nauplii used in each experiment were obtained from eggs hatched from a single female. Healthy nauplii were stocked in conical fiberglass tanks containing 270 1 of filtered seawater provided with aeration at a density of 90-100/l. The larvae were reared from the Z1 to M3 stage for a total of 8 days using either Tetraselmis chuii or Chaetoceros calcitrans. The treatments were arranged with six replicates for each algal food and placed at random. Three experimental runs were conducted. T. chuii and C. calcitruns were cultured in “F” medium (Guillard and Ryther, 1962) two days prior to feeding. Cultures were harvested during the exponential growth phase and fed to the larvae. Water samples in the larval tanks were examined three times daily under the microscope with a hemacytometer to determine cell density. A feeding density of 50-100 X lo3 cells/ml was maintained either by draining and adding filtered seawater or by adding cultured phytoplankton. Newly hatched brine shrimp nauplii (San Francisco Bay variety) were fed to the larvae during late M? stage. A level of 2-5 nauplii/ml was maintained. About 30-50% of the total volume of seawater was changed daily, from the fourth to the eight day of culture. Water exchange was made before feeding and after physicochemical parameters, such as water temperature, salinity, pH and ammonia content, had been monitored. Daily estimates of larval population were made by counting the larvae in four aliquot samples. Growth, measured in terms of the developmental stage, was recorded daily by taking two batches of 10 larvae from each treatment. The larvae were examined under the microscope and identified following the classification of Villaluz et al. (1969). Growth was quantified by the growth index of the larvae (Villegas and Kanazawa, 1979): Growth (developmental stage) = A/10. Where A = absolute value X number of larvae. The absolute value is established as follows: Larval stage
Absolute value
Zoea
1 2 3 4 5 6
Mysis
1 2 3 1 2 3
c
For each treatment, larvae from three tanks were harvested at Z3 stage and the remaining three tanks at M3 for proximate analysis. Before analysis,
255
the larvae were drained in a hand net, washed with distilled water twice, oven dried at 105°C and stored in bottles. Crude protein was determined by the semi-micro-Kjeldahl distillation method (Pearson, 1977), and lipids were extracted with chloroform-methanol-water (Bligh and Dyer, 1959). Ash was determined by the method described by Schneider and Flatt (1975). First the method of Maynard (1970) for crude fiber was followed then percentage dry matter of the samples was determined by oven-drying at 105°C to constant weight for 2-5 h. Carbohydrate values were obtained by difference. Duplicate samples were taken for each analysis. The chemical compositions of C. calcitruns and T. chuii were obtained from Benitez et al. (unpublished). The proximate analysis of newly hatched brine shrimp followed the procedures for P. monodon larvae. RESULTS AND DISCUSSION
Survival and growth A sharp drop in the population of larvae fed T. chuii was observed on the second and third days, from the fourth day to the seventh day; the decrease was about 6% (Table I). Survival of larvae fed C. calcitruns decreased gradually during the zoeal stages and dropped abruptly from the fourth to the sixth day. F-test showed significant differences in mean percentage survival between C. calcitrans and T. chuii-fed larvae on the second and third days, but not the succeeding days (Table I). Similar results were obtained on growth (Table II). The differences in survival and growth rates during the second and third TABLE I Mean percentage survival and F values of P. monodon and T. chuii Day
1 2 3 4 5 6 7 8
larvae (Z,-M,)
Treatment Larvae fed C. calcitrans
Larvae fed T. chuii
100.0 94.0 88.9 80.4 68.1 58.7 51.9 47.1
100.0 * 0.00
aTabular F value: F: (0.05) = 7.71 (0.01) = 21.20
* * f f f f + +
0.00 3.92 4.54 6.86 3.15 2.82 2.76 0.31
85.6 78.0 70.8 64.0 58.1 52.9 50.2
f 1.63 + 4.66 f 7.75 ? 6.55 +_6.45 k 3.41 k 3.78
P values 39.65** 8.48* 2.58 0.94 0.03 0.15 2.01
fed C. calcitrans
256 TABLE II Mean growth (absolute values) and F. values of P. monodon C. calcitrans and T. chuii Day
1 2 3 4 5 6 I 8
larvae (Z,-M,)
fed
Treatment Larvae fed C. calcitrans
Larvae fed T. chuii
Fa values
1.0 2.0 2.9 3.0 3.5 4.3 5.4 5.9
1.0 1.7 2.7 3.0 3.9 4.5 5.5 5.9
61.33** 13.25* 2.0 1.92 0.15 0.02 0.15
f 0.00 * 0.00 ? 0.10 ? 0.00 i 0.35 It 0.38 f 0.52 i 0.23
f; f + f f t f i
0.00 0.06 0.00 0.00 0.23 0.42 0.49 0.21
-
aTabular F value: F: (0.05) = 7.71 (0.01) = 21.20
days can probably be attributed to cell size. Cells of C. calcitruns, although bearing setae of considerable length, measure 4-5 /_m, while T. chuii cells are 12-15 pm. Although measurements of the mouths of the early zoeal stage (Z, and Z,) of P. monodon were not made, the-mouths are probably not wide enough to take in cells of T. chuii. Consequently, the larvae could not ingest enough food. Particle size is an important consideration in the rearing of larval forms of animals (Frost, 1972). Jones et al. (1979) found 10 pm to be the optimum size for Z1 stage of P. japonicus fed a microencapsulated diet. Yang (1975) reported that the Z1 stage of P. juponicus was initially capable of ingesting food particles in the range of 3-5 m. At Z,, the mouth of the larvae could take in cells larger than 200 pm. Similar results were obtained in this experiment. There was a trend towards selection of larger particles in later zoea and mysis stages. Macronutrient
composition
of food and larvae
The general composition of T. chuii, C. calcitrans and A. salina nauplii are shown in Table III. The chemical composition of the two algal species applies to the growing cells in the exponential phase of growth. Differences in protein content at Z3 and MJ of P. monodon larvae fed T. chuii and C. calcitruns were significant (1%) using the F-test as shown in Table IV. T. chuii, which contains more protein than C. calcitrans, significantly influenced the protein content of the larvae. However, growth and survival was significantly better in larvae fed C. calcitruns than in larvae fed T. chuii.
257 TABLE
III
Average
percentage
macronutrient
composition
of T. chuii, C. calcitrans and A. salina
Food
Crude protein
Crude lipid
Carbohydratea
C. calcitransb T. chuii A. salina
23.94’ 49.22 55.21
8.69 10.60 12.53
19.01 16.38 18.06
aObtained by difference. bBenitez et al. (unpublished). TABLE
IV
Means and F values of macronutrient and C. calcitrans Macronutrient (% dry weight)
Crude protein at Z, M, Lipid at Z, M, Carbohydrateb at Z, M,
composition
of Z, and M, larvae fed T. chuii
Treatment Larvae fed C. calcitrans
Larvae fed T. chuii
F= values
35.68 f 0 32 49.71 f 0.34
41.65 f 0.57 51.05 f 0.17
247.02** 35.95**
0.25 f 0.03 0.46 f 0.4
0.08 f 0.03 0.35 j- 0.03
52.89** 13.31*
48.28 f 0.61 27.48 2 0.68
47.23 f 0.71 33.50 ? 0.47
3.82 160.47**
aTabular F value: F; (0.05) = 7.71 (0.01) = 21.20. bObtained by difference.
It is possible that the amino acid pattern in C. calcitruns was better than that in T. chuii. It is not only the quantity but also the quality of protein that enhances growth and survival (Kakade, 1974). The addition of A. salina could have improved the amino acid as well as the fatty acid pattern of the food. Generally, two or more sources of protein are better than one because of the possible improvement of amino acid pattern of the protein (Rumsey and Ketola, 1975). Lipid content of C. calcitruns-fed larvae was significantly higher, using the F-test, at Z3 and M3 than those given T. chuii. As discussed earlier, possibly more C. culcitruns than T. chuii were ingested because of the smaller cell size, thus causing the difference in lipid content in the larvae. Similarly in P. setiferus larvae, Ward et al. (1979) observed that major fatty acids increase significantly from zoea to l-2 day postlarvae. New (1976) reported that specific
258
fatty acids have been shown to be more important than total lipid content of the diet of the shrimps. Although the fatty acid contents of the phytoplankton were not analyzed, it is inferred that the fatty acid quality of C. calcitrans was better than that of T. chuii, because of the higher growth and survival in Chaetoceros-fed larvae compared with Te trasetmis-fed larvae. Carbohydrate content at Z3 was slightly higher in C. calcitrans-fed larvae than in T. chuii-fed larvae. Although there was a decrease in carbohydrate content of the larvae at M2 stage in both treatments, significantly higher values were obtained for T. chuii larvae than C. calcitrans larvae. Physicochemical parameters Ranges and means of the physicochemical parameters were monitored daily during the culture period. There was little variation in water temperature (26.5-2&8”C), salinity (30.5-32.7°/oo) and pH (8.2-8.3) of the rearing medium in both treatments. Levels of NH3-N are given in Table V. High ammonia levels were measured in both treatments on the fourth day of culture. Water exchange was begun on the fourth day after monitoring of physicochemical parameters. The increase in ammonia concentration up to the fourth day was probably due to the accumulation of larval feces in the medium. According to Kinne (1976), ammonia is the chief excretory product of crustaceans. TABLE V Means of NH, -N levels. Values presented are based on daily average of three trials Day
ppm (mg NH,-N/l)
C. calcitrans 1 2 3 4 5 6 7 8
0.015 0.053 0.112 0.340 0.095 0.234 0.084 0.120
+_0.01 + 0.02 + 0.01 f 0.05 f 0.03 2 0.03 + 0.03 t 0.02
T. chuii 1 2 3 4 5 6 7 8
0.012 0.038 0.061 0.289 0.095 0.234 0.070 0.130
+ 2 ? + ? * f f
0.01 0.01 0.02 0.03 0.03 0.03 0.02 0.01
259
Mean ammonia concentration during the fourth day of culture was higher in the C. calcitrans- than in the T. chuii-fed larval tanks. Differences are attributed to the accummulation of feces from a denser population brought about by the higher survival rate. Although high concentrations were recorded from the second to fourth day, this did not affect the survival and growth of the larvae, as shown by the high survival rate obtained in C. calcitruns-fed larvae. Catedral et al. (1977) reported that P. monodon larvae can tolerate up to 10 NH3 -N mg/l. ACKNOWLEDGEMENT
The work reported in this article is based on a thesis submitted by the senior author in partial fulfillment of the requirements for the MS. Fisheries (Aquaculture) degree from the University of the Philippines System. The thesis project was financed by the MNR/BFAR/SEAFDEC/PCARR Graduate Study Grant. Thanks are due to Engr. Rolando Platon for guidance during the experimental phase of the study and J.H. Primavera for encouragement. Contribution
No. 105 of the SEAFDEC,
Aquaculture
Department.
REFERENCES Aquacop, 1977. Elevage larvaire de Peneides en milieu tropical Groupe de travail aquaculture I.C.E.S. 3rd meeting. Working group on mariculture, Brest, France. Actes Colloq. C.N.E.X.O., 4: 179-191. Bligh, E.G. and Dyer, W.J., 1959. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol., 37: 911-917. Benitez, L.V., Millamena, 0. and Penaflorida, R., 1980. The determination of macronutrient composition of natural foods organic mass cultured in SEAFDEC, Aquaculture Department (Unpublished data). Catedral, F., Gerochi, D., Quibuyen, A. and Casalmir, C., 1977. Effect of nitrite, ammonia and temperature on P. monodon. SEAFDEC Aqua. Dept. Q. Res. Rpt. 1 (3): 9-12. Frost, W.B., 1972. Effects of size and concentration of food particles on the feeding behavior of the marine planktonic copepod, Colanus pacificus. Limnol. Oceanogr., 17: 805-815. Griffith, G.W., Kenslow, M.A. and Ross, L.A., 1973. A mass culture method for Tetraselmis sp., a promising food for larval crustaceans. Proc. World Maricult. Sot., 4: 289294. Guillard, R.R.L. and Ryther, R.H., 1962. Studies on marine planktonic diatoms. I. Cyclotella nana Hdstedt and Denutola conferuacea (Cleve) Gran. Can. J. Microbial., 8: 229-239. Heinen, J.M., 1976. An introduction to culture methods for larval and postlarval penaeid shrimp. Proc. World Maricult. Sot. 7: 333-344. Hirata, H., Mori, Y. and Watanabe, M., 1975. Rearing of prawn larvae, Penaeus japonicus, fed soy-cake particles and diatoms. Mar. Biol., 2: 9-l 3. Jones, D.A., Kanazawa, A. and Abdel Rahman, S., 1979. Studies on the presentation of artificial diets for rearing the larvae P. japonicus Bate. Aquaculture, 17: 33-43. Kakade, M.L., 1974. Biochemical basis for the differences in plant protein utilization. J. Agric. Food Chem., 22: 550-555.
260 Kinne, O., 1976. Cultivation of marine organisms: water quality management and technology. In: 0. Kinne (Editor), Mar. Ecol. Wiley Interscience, New York, 3( 1): 79-300. Maynard, J., 1970. Methods of Food Analysis, Academic Press, New York, NY, 163 pp. New, M.B., 1976. A review of dietary studies with shrimp and prawns. Aquaculture, 9: 101-144. Pearson, D., 1977. The Chemical Analysis of Foods, seventh edition. Chem. Publishing Co., Inc., New York, NY, 575 pp. Rumsey, G.L. and Ketola, H.C., 1975. Amino acid supplementation of casein in diets of Atlantic salmon (Salmo salar) fry and of soybean meal on rainbow trout (Salmo gairdneri) fingerlings. J. Fish. Res. Board Can., 32: 422-426. Schneider, B.H. and Flatt, W.P., 1975. Evaluation of Feeds through Digestibility Experiments. Univ. of Georgia Press, 423 pp. Simon, C.M., 1978. The culture of diatom Chaetoceros gracilis and its use as a food for penaeid protozoeal larvae. Aquaculture, 14: 105-113. Villaluz, D.K., Villaluz, A.C., Ladrera, B., Sheik, M. and Gonzaga, M., 1969. Reproduction larval development and cultivation of sugpo (P. monodon Fabricius). Philipp. J. Sci., 90: 205-236. Villegas, C.T. and Kanazawa, A., 1979. Relationship between diet and growth weight of zoeal and mysis stages of I? japonicus Bate. Fish. Res. J. Philipp., 4: 32-40. Ward, D.G., Middleditch, B.S., Missler, S.R. and Lawrence, A.L., 1979. Fatty acid changes during larval development of Penaeus setiferus. Proc. World Maricult. Sot., 10: 464471. Yang, W.T., 1975. A manual for large-tank culture of penaeid shrimp to the postlarval stages. Miami, FL. Sea Grant. Tech. Bull. No. 31. Univ. of Miami, 68 pp.
253 Amsterdam
- Printed
in The Netherlands
GROWTH, SURVIVAL AND MACRONUTRIENT COMPOSITION OF PENAEUS MONODON FABRICIUS LARVAE FED WITH CHAETOCEROS CALCITRANS AND TETRASELMIS CHUII
EMILIA
TOBIAS-QUINITIO
Aquaculture Department, Iloilo (The Philippines) (Accepted
12 December
and CESAR
T. VILLEGAS
Southeast Asian Fisheries Development
Center, P.O. Box 256,
1981)
ABSTRACT Tobias-Quinitio, E. and Villegas, C.T., 1982. Growth, survival and macronutrient composition of Penaeus monodon Fabricius larvae fed with Chaetoceros calcitrans and Tetraselmis chuii. Aquaculture, 29: 253-260. Penaeus monodon larvae were reared from zoea, (Z,) to mysis, (M,) using two different algal feeds, Chaetoceros calcitrans and Tetraselmis chuii. Artemia nauplii were added to both treatments at mysis, . Mean survival and growth rates in both treatments were different at 5% level of significance on the second and third day of culture, but did not differ during the mysis stage and the end of the g-day culture period. Z, and M, larvae fed with C. calcitrans had a lower crude protein but a higher lipid content than T. chuii-fed larvae. Differences in carbohydrate content were noticed in M, larvae. The implication of the findings are discussed. INTRODUCTION
Various species of phytoplankton have been used as food in the culture of penaeid larvae (Simon, 1978). Griffith et al. (1973) showed that the growth of P. setiferus and P. aztecus larvae was more rapid with Tetraselmis than Skeletonema. The use of the diatom, Chaetoceros grucilis as exclusive food for P. styEirostris and P. uannamei zoea (Z) to mysis (M) was effective in obtaining a survival rate of 84.8% (Simon, 1978). Other genera of phytoplankton such as Isochrysis, Cylindrotheca (Aquacop, 1977), Phaeodactylum (Heinen, 1976) and Nitzschia (Hirata et al., 1975) have been used in penaeid culture. Some increased survival and growth rates of the larvae, while others gave good growth but poor survival, and vice versa. These differences may be due to size or chemical composition of algal cells. There is a continuous need to evaluate the effects of the algal food on the survival, growth and chemical composition of the prawn in order to establish good working criteria for the production of P. monodon postlarvae. This study investigates the effects of two species of phytoplankton, the diatom Chaetoceros calcitruns and the flagellate Tetruselmis chuii, on the growth and survival
0044-8486/82/0000-0000/$02.75
0 1982
Elsevier
Scientific
Publishing
Company
254
of P. monodon; compares the nutritive values of C. calcitruns and T. chuii, and the effects of both on the chemical composition of whole larvae at the zoea and mysis stages. MATERIALS
AND METHODS
P. monodon nauplii used in each experiment were obtained from eggs hatched from a single female. Healthy nauplii were stocked in conical fiberglass tanks containing 270 1 of filtered seawater provided with aeration at a density of 90-100/l. The larvae were reared from the Z1 to M3 stage for a total of 8 days using either Tetraselmis chuii or Chaetoceros calcitrans. The treatments were arranged with six replicates for each algal food and placed at random. Three experimental runs were conducted. T. chuii and C. calcitruns were cultured in “F” medium (Guillard and Ryther, 1962) two days prior to feeding. Cultures were harvested during the exponential growth phase and fed to the larvae. Water samples in the larval tanks were examined three times daily under the microscope with a hemacytometer to determine cell density. A feeding density of 50-100 X lo3 cells/ml was maintained either by draining and adding filtered seawater or by adding cultured phytoplankton. Newly hatched brine shrimp nauplii (San Francisco Bay variety) were fed to the larvae during late M? stage. A level of 2-5 nauplii/ml was maintained. About 30-50% of the total volume of seawater was changed daily, from the fourth to the eight day of culture. Water exchange was made before feeding and after physicochemical parameters, such as water temperature, salinity, pH and ammonia content, had been monitored. Daily estimates of larval population were made by counting the larvae in four aliquot samples. Growth, measured in terms of the developmental stage, was recorded daily by taking two batches of 10 larvae from each treatment. The larvae were examined under the microscope and identified following the classification of Villaluz et al. (1969). Growth was quantified by the growth index of the larvae (Villegas and Kanazawa, 1979): Growth (developmental stage) = A/10. Where A = absolute value X number of larvae. The absolute value is established as follows: Larval stage
Absolute value
Zoea
1 2 3 4 5 6
Mysis
1 2 3 1 2 3
c
For each treatment, larvae from three tanks were harvested at Z3 stage and the remaining three tanks at M3 for proximate analysis. Before analysis,
255
the larvae were drained in a hand net, washed with distilled water twice, oven dried at 105°C and stored in bottles. Crude protein was determined by the semi-micro-Kjeldahl distillation method (Pearson, 1977), and lipids were extracted with chloroform-methanol-water (Bligh and Dyer, 1959). Ash was determined by the method described by Schneider and Flatt (1975). First the method of Maynard (1970) for crude fiber was followed then percentage dry matter of the samples was determined by oven-drying at 105°C to constant weight for 2-5 h. Carbohydrate values were obtained by difference. Duplicate samples were taken for each analysis. The chemical compositions of C. calcitruns and T. chuii were obtained from Benitez et al. (unpublished). The proximate analysis of newly hatched brine shrimp followed the procedures for P. monodon larvae. RESULTS AND DISCUSSION
Survival and growth A sharp drop in the population of larvae fed T. chuii was observed on the second and third days, from the fourth day to the seventh day; the decrease was about 6% (Table I). Survival of larvae fed C. calcitruns decreased gradually during the zoeal stages and dropped abruptly from the fourth to the sixth day. F-test showed significant differences in mean percentage survival between C. calcitrans and T. chuii-fed larvae on the second and third days, but not the succeeding days (Table I). Similar results were obtained on growth (Table II). The differences in survival and growth rates during the second and third TABLE I Mean percentage survival and F values of P. monodon and T. chuii Day
1 2 3 4 5 6 7 8
larvae (Z,-M,)
Treatment Larvae fed C. calcitrans
Larvae fed T. chuii
100.0 94.0 88.9 80.4 68.1 58.7 51.9 47.1
100.0 * 0.00
aTabular F value: F: (0.05) = 7.71 (0.01) = 21.20
* * f f f f + +
0.00 3.92 4.54 6.86 3.15 2.82 2.76 0.31
85.6 78.0 70.8 64.0 58.1 52.9 50.2
f 1.63 + 4.66 f 7.75 ? 6.55 +_6.45 k 3.41 k 3.78
P values 39.65** 8.48* 2.58 0.94 0.03 0.15 2.01
fed C. calcitrans
256 TABLE II Mean growth (absolute values) and F. values of P. monodon C. calcitrans and T. chuii Day
1 2 3 4 5 6 I 8
larvae (Z,-M,)
fed
Treatment Larvae fed C. calcitrans
Larvae fed T. chuii
Fa values
1.0 2.0 2.9 3.0 3.5 4.3 5.4 5.9
1.0 1.7 2.7 3.0 3.9 4.5 5.5 5.9
61.33** 13.25* 2.0 1.92 0.15 0.02 0.15
f 0.00 * 0.00 ? 0.10 ? 0.00 i 0.35 It 0.38 f 0.52 i 0.23
f; f + f f t f i
0.00 0.06 0.00 0.00 0.23 0.42 0.49 0.21
-
aTabular F value: F: (0.05) = 7.71 (0.01) = 21.20
days can probably be attributed to cell size. Cells of C. calcitruns, although bearing setae of considerable length, measure 4-5 /_m, while T. chuii cells are 12-15 pm. Although measurements of the mouths of the early zoeal stage (Z, and Z,) of P. monodon were not made, the-mouths are probably not wide enough to take in cells of T. chuii. Consequently, the larvae could not ingest enough food. Particle size is an important consideration in the rearing of larval forms of animals (Frost, 1972). Jones et al. (1979) found 10 pm to be the optimum size for Z1 stage of P. japonicus fed a microencapsulated diet. Yang (1975) reported that the Z1 stage of P. juponicus was initially capable of ingesting food particles in the range of 3-5 m. At Z,, the mouth of the larvae could take in cells larger than 200 pm. Similar results were obtained in this experiment. There was a trend towards selection of larger particles in later zoea and mysis stages. Macronutrient
composition
of food and larvae
The general composition of T. chuii, C. calcitrans and A. salina nauplii are shown in Table III. The chemical composition of the two algal species applies to the growing cells in the exponential phase of growth. Differences in protein content at Z3 and MJ of P. monodon larvae fed T. chuii and C. calcitruns were significant (1%) using the F-test as shown in Table IV. T. chuii, which contains more protein than C. calcitrans, significantly influenced the protein content of the larvae. However, growth and survival was significantly better in larvae fed C. calcitruns than in larvae fed T. chuii.
257 TABLE
III
Average
percentage
macronutrient
composition
of T. chuii, C. calcitrans and A. salina
Food
Crude protein
Crude lipid
Carbohydratea
C. calcitransb T. chuii A. salina
23.94’ 49.22 55.21
8.69 10.60 12.53
19.01 16.38 18.06
aObtained by difference. bBenitez et al. (unpublished). TABLE
IV
Means and F values of macronutrient and C. calcitrans Macronutrient (% dry weight)
Crude protein at Z, M, Lipid at Z, M, Carbohydrateb at Z, M,
composition
of Z, and M, larvae fed T. chuii
Treatment Larvae fed C. calcitrans
Larvae fed T. chuii
F= values
35.68 f 0 32 49.71 f 0.34
41.65 f 0.57 51.05 f 0.17
247.02** 35.95**
0.25 f 0.03 0.46 f 0.4
0.08 f 0.03 0.35 j- 0.03
52.89** 13.31*
48.28 f 0.61 27.48 2 0.68
47.23 f 0.71 33.50 ? 0.47
3.82 160.47**
aTabular F value: F; (0.05) = 7.71 (0.01) = 21.20. bObtained by difference.
It is possible that the amino acid pattern in C. calcitruns was better than that in T. chuii. It is not only the quantity but also the quality of protein that enhances growth and survival (Kakade, 1974). The addition of A. salina could have improved the amino acid as well as the fatty acid pattern of the food. Generally, two or more sources of protein are better than one because of the possible improvement of amino acid pattern of the protein (Rumsey and Ketola, 1975). Lipid content of C. calcitruns-fed larvae was significantly higher, using the F-test, at Z3 and M3 than those given T. chuii. As discussed earlier, possibly more C. culcitruns than T. chuii were ingested because of the smaller cell size, thus causing the difference in lipid content in the larvae. Similarly in P. setiferus larvae, Ward et al. (1979) observed that major fatty acids increase significantly from zoea to l-2 day postlarvae. New (1976) reported that specific
258
fatty acids have been shown to be more important than total lipid content of the diet of the shrimps. Although the fatty acid contents of the phytoplankton were not analyzed, it is inferred that the fatty acid quality of C. calcitrans was better than that of T. chuii, because of the higher growth and survival in Chaetoceros-fed larvae compared with Te trasetmis-fed larvae. Carbohydrate content at Z3 was slightly higher in C. calcitrans-fed larvae than in T. chuii-fed larvae. Although there was a decrease in carbohydrate content of the larvae at M2 stage in both treatments, significantly higher values were obtained for T. chuii larvae than C. calcitrans larvae. Physicochemical parameters Ranges and means of the physicochemical parameters were monitored daily during the culture period. There was little variation in water temperature (26.5-2&8”C), salinity (30.5-32.7°/oo) and pH (8.2-8.3) of the rearing medium in both treatments. Levels of NH3-N are given in Table V. High ammonia levels were measured in both treatments on the fourth day of culture. Water exchange was begun on the fourth day after monitoring of physicochemical parameters. The increase in ammonia concentration up to the fourth day was probably due to the accumulation of larval feces in the medium. According to Kinne (1976), ammonia is the chief excretory product of crustaceans. TABLE V Means of NH, -N levels. Values presented are based on daily average of three trials Day
ppm (mg NH,-N/l)
C. calcitrans 1 2 3 4 5 6 7 8
0.015 0.053 0.112 0.340 0.095 0.234 0.084 0.120
+_0.01 + 0.02 + 0.01 f 0.05 f 0.03 2 0.03 + 0.03 t 0.02
T. chuii 1 2 3 4 5 6 7 8
0.012 0.038 0.061 0.289 0.095 0.234 0.070 0.130
+ 2 ? + ? * f f
0.01 0.01 0.02 0.03 0.03 0.03 0.02 0.01
259
Mean ammonia concentration during the fourth day of culture was higher in the C. calcitrans- than in the T. chuii-fed larval tanks. Differences are attributed to the accummulation of feces from a denser population brought about by the higher survival rate. Although high concentrations were recorded from the second to fourth day, this did not affect the survival and growth of the larvae, as shown by the high survival rate obtained in C. calcitruns-fed larvae. Catedral et al. (1977) reported that P. monodon larvae can tolerate up to 10 NH3 -N mg/l. ACKNOWLEDGEMENT
The work reported in this article is based on a thesis submitted by the senior author in partial fulfillment of the requirements for the MS. Fisheries (Aquaculture) degree from the University of the Philippines System. The thesis project was financed by the MNR/BFAR/SEAFDEC/PCARR Graduate Study Grant. Thanks are due to Engr. Rolando Platon for guidance during the experimental phase of the study and J.H. Primavera for encouragement. Contribution
No. 105 of the SEAFDEC,
Aquaculture
Department.
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