- Email: [email protected]
Culture of the calanoid copepod Pseudodiaptomus euryhalinus (Johnson 1939) with different microalgal diets
Aquaculture 290 (2009) 317–319
Contents lists available at ScienceDirect
Aquaculture j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / a q u a - o n l i n e
Culture of the calanoid copepod Pseudodiaptomus euryhalinus (Johnson 1939) with different microalgal diets A.C. Puello-Cruz a, S. Mezo-Villalobos a, B. González-Rodríguez a, D. Voltolina b,⁎ a b
Centro de Investigaciones en Alimentación y Desarrollo A.C., Unidad Mazatlán, Sinaloa, Mexico Centro de Investigaciones Biológicas del Noroeste, Laboratorio UAS-CIBNOR, P.O. Box 1132, Mazatlán, Sinaloa, Mexico
a r t i c l e
i n f o
Article history: Received 16 June 2008 Received in revised form 14 February 2009 Accepted 16 February 2009 Keywords: Copepod culture Yield Monoalgal diets Mixed diets
a b s t r a c t The copepod Pseudodiaptomus euryhalinus was fed 320 cells μL− 1 of monoalgal cultures of Chaetoceros muelleri, Nannochloropsis oculata, Isochrysis galbana, Tetraselmis suecica, or a commercial frozen concentrate of Tetraselmis sp., and the diet which gave the best production was compared in a second experiment to three mixed diets: C. muelleri:I. galbana supplied in 1:1 and 2:1 cell ratios and C. muelleri:I. galbana:frozen Tetraselmis sp. in 2:2:1 ratio. These gave better results than the cultures of N. oculata, I. galbana, T. suecica and the frozen Tetraselmis concentrate, but the production was similar to that obtained with C. muelleri supplied as a monoalgal diet, showing that the addition of C. muelleri may improve the performance of other monoalgal diets, whereas the addition of other microalgae is unlikely to improve the results obtained when C. muelleri is supplied as a monoalgal diet. © 2009 Elsevier B.V. All rights reserved.
1. Introduction Several studies have shown that copepods have a higher nutritional value than other live food sources for many fish larvae, because their broad size range, high digestibility and high food value allow better growth and development than other live diets (Støttrup and Norsker, 1997; McKinnon et al., 2003; Rippingale and Payne, 2001). In addition, calanoids are planktonic and have a discontinuous swimming behaviour, which is an important visual stimulus for marine fish larvae (Delbare et al., 1996; Schipp, 2006). However, the techniques for copepod mass culture are at their early stage of development and in particular little is known on the diets that might allow large-scale cultures for sustained and reliable biomass production. Most calanoid copepods are filter-feeders, but there is little information on their natural food preference and on the individual food value of the species present in natural phytoplankton. For this reason, the identification of microalgal diets that allow good survival, fast rates of somatic growth and of reproduction as well as high copepod yields is of paramount importance for larval fish culture. There is a wide range of microalgae available for aquaculture, mostly chosen for their specific nutritional qualities and because they may be easily grown in mass cultures. Some of the most widely
⁎ Corresponding author. Tel.: +52 669 981 03 77; fax: +52 669 982 86 56. E-mail address: [email protected] (D. Voltolina). 0044-8486/$ – see front matter © 2009 Elsevier B.V. All rights reserved. doi:10.1016/j.aquaculture.2009.02.016
used in commercial aquaculture are Chaetoceros muelleri Lemmerman, Isochrysis galbana Parke, Tetraselmis suecica (Kylin) Butcher and Nannochloropsis oculata (Droop) Hibberd (Coutteau, 1996; Duerr et al., 1998). However, monospecific diets may cause nutritional deficiencies, because of the inadequate content of one or more essential nutrients. To reduce this risk, several authors have suggested the use of mixed diets, because their combined nutrient contents are more likely to meet the nutritional requirements of the target species (Brown et al., 1989; Smith et al., 1992). In the case of copepods, this was confirmed by Milione et al. (2007), who found that the development rate of the calanoid Acartia sinjiensis was significantly better with a mixture of two microalgae than with monospecific diets. Semicontinuous cultures of Pseudodiaptomus euryhalinus Johnson fed a mixed algal diet (N. oculata, I. galbana, C. muelleri and T. suecica) have been routinely used in our laboratory since several years to feed the early larval stages of the native puffer fish Sphoeroides annulatus Jenyns and of the red snapper Lutjanus guttatus (Steindachner) (García-Ortega, 2009). The aim of this work was to determine if it was possible to simplify the culture procedure using monoalgal diets, or to improve the productivity of our P. euryhalinus cultures with simpler mixed diets. The feeding rates of filter-feeders may be modified by the external particle concentration or by the volume of the food particles available or ingested (Støttrup and Jensen, 1990; Yúfera and Lubián, 1990), but models based on food availability or carbon ingestion are inadequate to explain copepod egg production (Ohman and Runge, 1994; Pond et al., 1996; Mayor et al., 2006). For this reason, in spite of the large
318
A.C. Puello-Cruz et al. / Aquaculture 290 (2009) 317–319
Table 1 Cell volume (μm3), ash-free dry weight and energy content (AFW in pg cell− 1 and J 10− 6 cell− 1) of the four microalgae used in this study.
a
Chaetoceros Isochrysisa Nannochloropsisa Tetraselmisb a b
μm3
AFW pg cell− 1
J cell− 1
140.2 35.3 8.2 329.4
34.8 17.1 6.3 89.2
0.745 0.496 0.134 1.99
Lora-Vilchis (2004). Piña et al. (2006).
differences in size, individual organic biomass and energy contents of the algal strains, the diets were supplied in equal cell numbers.
2. Materials and methods The 300 L cultures of P. euryhalinus of the Nutrition and Larviculture Laboratory of the Centro de Investigaciones en Alimentación y Desarrollo, Mazatlán Unit, are maintained at 35‰ salinity, 27 ± 1 °C and 12:12 h light:dark photoperiod, food is a 1:1:1:1 mixture in equal cell numbers of N. oculata, I. galbana, C. muelleri and T. suecica supplied daily in the appropriate volumes to maintain a near constant cell concentration of 320 cells μL− 1. Partial harvests (75%) are after two weeks, and the yields obtained with these cultures range between 1.46 and 3.93 organisms mL− 1. The experiments started placing five copulating pairs of P. euryhalinus with no egg sac in 1-L round bottom glass flasks with 500 mL of 5-μm filtered, UV-sterilized seawater (salinity 35 ± 1‰), with gentle aeration and 12:12 h light:dark photoperiod. The flasks were kept in a water bath to maintain temperature at 26 ± 1 °C. The first experiment served to evaluate the monoalgal diets C. muelleri, I. galbana, N. oculata, T. suecica (Table 1), and a commercial frozen concentrate of Tetraselmis sp. (similar in size to T. suecica); in the second experiment, the best treatment of experiment 1 was compared to three mixed diets formulated using C. muelleri, I. galbana and the frozen commercial product. C. muelleri was used as the control treatment, as well as in the mixed diets (1:1 and 2:1 cell number ratio with the second best diet I. galbana). The third was a mixture of C. muelleri + I. galbana + frozen Tetraselmis sp. (2:2:1, respectively). The feeding rates of filter-feeders may be modified by the external particle concentration or by the volume of the food particles available or ingested (Støttrup and Jensen, 1990; Yúfera and Lubián, 1990). However, it has been shown that models based on food availability or carbon ingestion are inadequate to explain copepod egg production (Ohman and Runge, 1994; Pond et al., 1996; Mayor et al., 2006). For this reason, in spite of the large differences in size, individual organic biomass and energy contents of the algal strains (Table 1), the diets were supplied in equal cell numbers. In all cases, each diet was evaluated in five replicate cultures. The algal concentrations were evaluated daily in each culture with a haemacytometer and the appropriate volumes of microalgae culture were added to maintain a stable daily food ration of 320 cells μL− 1.
After addition, the culture volume was corrected to 500 mL. Eighty percent of the seawater was renewed every third day. The experiments lasted 15 days. At the end, all organisms present in each culture were counted, separating ovigerous females, copulating pairs, copepodites and nauplii. After verification of the hypothesis of normality and equal variances, the mean total concentrations were compared using nonparametric or parametric ANOVA tests (first and second experiments) with α = 0.05, separating the different means with the respective Tukey's and Dunn's multiple comparison tests. 3. Results The mean total yield of the cultures fed C. muelleri was significantly higher than those obtained at the end of the first experiment with the other diets. I. galbana gave the second best result followed by the N. oculata and frozen Tetraselmis-fed cultures, which were not significantly different from each other but gave better results than those obtained with T. suecica (Table 2). These results were used to design the mixed diets for the second experiment. C. muelleri was used as the control treatment, as well as in the mixed diets (1:1 and 2:1 cell number ratio with the second best diet I. galbana). The third was a mixture of C. muelleri + I. galbana + frozen Tetraselmis sp. (2:2:1, respectively). The results of this second experiment showed that C. muelleri seems to support practically all the nutritional requirements of P. euryhalinus, since none of the mixtures could improve the nutritional value of monospecific C. muelleri cultures. The mean yields obtained in this experiment ranged from 462.92 ± 152.5 organism L− 1 (C. muelleri) to 296 ± 114.74 L− 1 with the mixture C. muelleri, I. galbana and frozen Tetraselmis sp. (Table 3), and there were no significant differences (P= 0.116) between diets. 4. Discussion The performance of a diet depends on its digestibility, composition and available energy content. Thus, the poor performance of the Nannochloropsis-based diet might be explained by the toughness and poor digestibility of its cell wall (Dhont and Lavens, 1996; Payne and Rippingale, 2000), or by its small size and low weight and energy content (close to 8.2 µm3, 6.3 pg and 0.134 10–6 J cell–1). However, size does not explain the poor results obtained with T. suecica, which are comparable to those found by Cripe (1997) and Piña et al. (2006) with Tetraselmis-fed penaeid shrimp larvae, and to the high degree of deformities and complete lack of development beyond the copepodite stage observed by Knuckey et al. (2005) in Tetraselmis-fed A. sinjiensis nauplii. In this case, digestibility might be the reason for the poor performance of this microalga, in view that production improved when it was supplied as a frozen and probably more digestible diet. Even in this case, this species gave mostly nauplii and copepodites with low adult yields, which could be due to the relatively low food value of this strain, or to the fact that frozen microalgae tend to adhere
Table 2 Production of Pseudodiaptomus euryhalinus with monospecific diets: TE F = Tetraselmis sp. commercial frozen concentrate; TE = Tetraselmis suecica; IS = Isochrysis galbana; CH = Chaetoceros muelleri; NN = Nannochloropsis oculata.
Table 3 Production (organisms L− 1) and stages of development (in %) of Pseudodiaptomus euryhalinus with C. muelleri supplied as a monospecific diet (control) and. with the mixed diets CI 1 = C. muelleri+I. galbana (1:1 ratio), CI 2 = C. muelleri+I. galbana (2:1 ratio) and CIT = C. muelleri+I. galbana + frozen Tetraselmis sp. (2:2:1 ratio).
Diet
TE F
TE
IS
CH
NN
Diet
Control
CI 1
CI 2
CIT
Org. L− 1 CPO CP CD Na
67.7 ± 36.7c – 2% 78% 20%
3 ± 2.9d – 1% 68% 31%
210.4 ± 137.3b 7% 6% 59% 28%
581.6 ± 333.7a 10% 13% 33% 44%
81.7 ± 44.3c 3% 10% 44% 43%
Org. L− 1 CPO CP CD Na
462.9 ± 152.5a 7% 10% 47% 36%
451.6 ± 177.8a 6% 11% 44% 39%
364 ± 167.5a 9% 13% 44% 34%
296 ± 114.7a 13% 17% 38% 32%
Different letters indicate significant difference (one way ANOVA, α = 0.05, a N b N c N d). CPO = copulating adults, ovigerous female; CP = copulating adults; CD = copepodites; Na = nauplii.
The equal letter indicates lack of significant difference (one way ANOVA, α = 0.05). CPO = copulating adults, ovigerous females; CP = copulating adults; CD = copepodites; Na = nauplii.
A.C. Puello-Cruz et al. / Aquaculture 290 (2009) 317–319
to swimming appendages, thus reducing the swimming and feeding activities of the organisms in culture (Paniagua-Chávez and Voltolina, 1995; Isiordia-Pérez and Puello-Cruz, 2007). C. muelleri and I. galbana are considered among the best food sources for filter-feeders, because of their high contents of essential fatty acids that promote high survival and growth (Watanabe et al., 1983; Brown et al., 1989). In the case of copepods, this has been confirmed by the high yields obtained by Støttrup and Jensen (1990) and Payne and Rippingale (2000) with these two microalgae supplied as monospecific diets. However, most authors agree on the high food value of C. muelleri and other single-celled Chaetoceros species for a wide variety of aquatic invertebrates (His and Robert, 1987; Cordero-Esquivel and Voltolina, 1994; Lora-Vilchis et al., 2004b; Suchar and Cigbu, 2006), indicating a better balance of the essential biochemical components of these species. In addition, several aquatic organisms fed with I. galbana have shown poor growth (Lora-Vilchis and Doktor, 2001; Lora-Vilchis et al., 2004a; Piña et al., 2006) and low survival (Helm and Laing, 1987; Cripe, 1997) indicating that, because of species-specific differences in dietary requirements, this species might be a poor source of some essential nutrient. For instance, on a mass to mass basis C. muelleri is a better source of ascorbic acid (Brown and Miller, 1992), as well as of the essential unsaturated fatty acids EPA and ARA (Coutteau, 1996; Piña et al., 2006), whereas I. galbana is richer in DHA. It has been shown that DHA, as well as n−3 fatty acids, may be converted into the essential EPA (Navarro et al., 1999), which may be taken as an indication of a high demand for this nutrient, since there is evidence that it is used as substrate by some copepod species (Veloza et al., 2006). The content of EPA of C. muelleri is close to five times higher than that of I. galbana (Lora-Vilchis et al., 2004a). This would cause a lower demand on the reserves of other fatty acids, which is the likely explanation for the good results we obtained with this diet. Mixed microalgae diets have been suggested for filter-feeders, because monoalgal cultures may not meet their nutritional requirements (Treece, 1984; Brown et al., 1989; Smith et al., 1992; Kleppel and Burkart, 1995). However, this is not a general rule (Yúfera and Lubián, 1990) and it was not the case in this study, because the production we obtained with one of the monospecific cultures, although lower than the 1 copepod mL− 1 mentioned by Payne and Rippingale (2000) for a different copepod species, could not be improved supplying different mixtures with other microalgae species. Acknowledgements Research was funded by the SAGARPA-CONACYT Project 2004C01-196. Gabriela Velasco supplied the microalgae cultures. References Brown, M.R., Miller, K.A., 1992. The ascorbic acid content of eleven species of microalgae used in mariculture. J. Appl. Phycol. 4, 205–215. Brown, M.R., Jeffrey, S.W., Garland, C.D., 1989. Nutritional Aspects of Microalgae Used in Mariculture; a Literature Review. Mar. Lab. Rep., vol. 205. CSIRO. 44 pp. Cordero-Esquivel, B., Voltolina, D., 1994. Growth of Mytilus galloprovincialis Lmk. with four microalgae and two feeding regimes. J. World Aquac. Soc. 25, 471–476. Coutteau, P., 1996. Micro-algae. In: Lavens, P., Sorgeloos, P. (Eds.), Manual on the Production And use of Live Food for Aquaculture. Fish. Tech. Pap., vol. 361. FAO, pp. 7–48. Cripe, G.M., 1997. Spawning and larval survival of the pink shrimp, Penaeus duorarum, in a small culture facility. J. Appl. Aquac. 7, 29–41. Delbare, D., Dhert, P., Lavens, P., 1996. Zooplankton. In: Lavens, P., Sorgeloos, P. (Eds.), Manual on the Production and Use of Live Food for Aquaculture. . Fish. Tech. Pap., vol. 361. FAO, pp. 252–282. Dhont, J., Lavens, P., 1996. Tank production and use of ongrown Artemia. In: Lavens, P., Sorgeloos, P. (Eds.), Manual on the Production and Use of Live Food for Aquaculture. Fish. Tech. Pap., vol. 361. FAO, pp. 164–195. Duerr, E.O., Molnar, A., Sato, V., 1998. Cultured microalgae as aquaculture feeds. J. Mar. Biotechnol. 7, 65–70.
319
García-Ortega, A., 2009. Nutrition and feeding research in the spotted rose snapper (Lutjanus guttatus) and bulleye puffer (Sphoeroides annulatus), new species for marine aquaculture. Fish Physiol. Biochem. 35, 69–80. Helm, M.M., Laing, I., 1987. Preliminary observations on the nutritional value of “Tahiti Isochrysis” to bivalve larvae. Aquaculture 62, 281–288. His, E., Robert, R., 1987. Croissance des larves de Crassostrea gigas et de Mytilus galloprovincialis en présence d'algues monocellulaires isolée du tractus digestif des veligéres du milieu naturel. Haliotis 16, 383–391. Isiordia-Pérez, E., Puello-Cruz, A., 2007. Evaluación del crecimiento y supervivencia en larvas de camarón blanco Litopenaeus vannamei usando como fuente de alimento microalgas vivas y congeladas. REDVET. Rev. Electrón. Vet., vol. 8(5). www. veterinaria.org/revistas/redvet/n070706.html (downloaded July 2008). Kleppel, G.S., Burkart, C.A., 1995. Egg Production and the Nutritional Environment of Acartia tonsa: the Role of Food Quality in Copepod Nutrition. J. Mar. Sci., 52. ICES, pp. 297–304. Knuckey, R.M., Semmens, G.L., Mayer, R.J., Rimmer, M.A., 2005. Development of an optimal microalgal diet for the culture of the calanoid copepod Acartia sinjiensis: effect of algal species and feed concentration on copepod development. Aquaculture 249, 339–351. Lora-Vilchis M.C. 2004. Evaluación de la calidad dietética de microalgas mediante el estudio del balance energético de Artemia franciscana. Ph. D. Thesis. Centro de Investigaciones Biológicas del Noroeste. La Paz, Mexico. 135 pp. Lora-Vilchis, M.C., Doktor, N., 2001. Evaluation of seven algal diets for spat of the pacific scallop Argopecten ventricosus. J. World Aquac. Soc. 32, 228–235. Lora-Vilchis, M.C., Cordero-Esquivel, B., Voltolina, D., 2004a. Growth of Artemia franciscana fed Isochrysis sp. and Chaetoceros muelleri during its early life stages. Aquac. Res. 35, 1086–1091. Lora-Vilchis, M.C., Ruiz-Velasco Cruz, E., Reynoso-Granados, T., Voltolina, D., 2004b. Evaluation of five microalgae diets for juvenile pen shells Atrina maura (Sowerby, 1835). J. World Aquac. Soc. 35, 232–236. Mayor, D.H.J., Anderson, T.R., Irigoien, X., Harris, R., 2006. Feeding and reproduction of Calanus finmarchicus during non-bloom conditions in the Irminger Sea. J. Plankton Res. 28, 1167–1179. McKinnon, A.D., Duggan, S., Nichols, P.D., Rimmer, M.A., Semmens, G., Robino, B., 2003. The potential of tropical paracalanoid copepods as live feeds in aquaculture. Aquaculture 223, 89–106. Milione, M., Zeng, C., Tropical Crustacean Aquaculture Research Group, 2007. The effects of algal diets on population growth and egg hatching success of the tropical calanoid copepod, Acartia sinjiensis. Aquaculture 273, 656–664. Navarro, J.C., Henderson, R.J., McEvoy, L.A., Bell, M.B., Amat, F., 1999. Lipid conversion during enrichment of Artemia. Aquaculture 174, 155–166. Ohman, M.D., Runge, J.A., 1994. Sustained fecundity when phytoplankton resources are in short supply: omnivory by Calanus finmarchicus in the Gulf of St. Lawrence. Limnol. Oceanogr. 39, 21–36. Paniagua-Chávez, C.G., Voltolina, D., 1995. Fresh and frozen Dunaliella sp. (Chlorophyceae, Volvocales) as feed for Artemia franciscana Kellogg (Crustacea, Branchiopoda). Riv. Ital. Acquac. 30, 19–23. Payne, M.F., Rippingale, R.J., 2000. Evaluation of diets for culture of the calanoid copepod Gladioferens imparipes. Aquaculture 187, 85–96. Piña, P., Voltolina, D., Nieves, M., Robles, M., 2006. Survival, development and growth of the Pacific white shrimp Litopenaeus vannamei protozoea larvae, fed monoalgal and mixed diets. Aquaculture 253, 523–530. Pond, D., Harris, R.P., Head, R.N., Harbour, D., 1996. Environmental and nutritional factors determining seasonal variability in the fecundity and egg viability of Calanus helgolandicus in coastal waters off Plymouth, U.K. Mar. Ecol., Prog. Ser. 143, 45–63. Rippingale, R.J., Payne, M.F., 2001. Intensive cultivation of a calanoid copepod Gladioferens imparipes. A Guide to Procedures. Dept of Environmental Biology. Curtin University of Technology. Schipp, G., 2006. The use of calanoid copepods in semi-intensive tropical marine fish larviculture. In: Cruz-Suárez, L.E., Rique-Marie, D., Tapia-Salazar, M., Nieto-López, M.G., Villareal-Cavazos, D.A., Puello-Cruz, A.C., García-Ortega, A. (Eds.), Avances en Nutrición Acuícola VIII. 8th Int. Symp. Aquatic Nutrition. Univ. Autón. Nuevo León, Monterrey. 84–94 pp. Smith, L.L., Biedenbach, J.M., Lawrence, A.L., 1992. Penaeid larviculture: Galveston method. In: Fast, A.E., Lester, L.J. (Eds.), Marine Shrimp Culture, Principles and Practices. Elsevier Science, Amsterdam, pp. 171–191. Støttrup, J.G., Jensen, J., 1990. Influence of algal diet on feeding and egg-production of the calanoid copepod Acartia tonsa Dana. J. Exp. Mar. Biol. Ecol. 141, 87–105. Støttrup, J.G., Norsker, N.H., 1997. Production and use of copepods in marine fish larviculture. Aquaculture 155, 231–247. Suchar, V.A., Chigbu, P., 2006. The effects of algae species and densities on the population growth of the marine rotifer, Colurella dicentra. J. Exp. Mar. Biol. Ecol. 337, 96–102. Treece, G.D., 1984. Larval rearing technology. In: Chamberlain, G.M., Haby, M.G., Miget, R.J. (Eds.), Texas Shrimp Farming Manual: an Update on Current Technology. Texas Agricultural Extension Service, Texas A & M University, Corpus Christi, pp. 43–64. Veloza, A., Chu, F.-L., Tang, K.W., 2006. Trophic modification of essential fatty acids by heterotrophic protists and its effects on the fatty acid composition of the copepod Acartia tonsa. Mar. Biol. 148, 779–788. Watanabe, T., Katajima, C., Fujita, S., 1983. Nutritional value or live organisms used in Japan for mass propagation of fish: a review. Aquaculture 34, 115–143. Yúfera, M., Lubián, L.M., 1990. Effects of microalgal diet on growth and development of invertebrates in marine aquaculture. In: Akatsuka, I. (Ed.), Introduction to Applied Phycology. Academic Publishing, The Hague. 209–227 pp.
Contents lists available at ScienceDirect
Aquaculture j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / a q u a - o n l i n e
Culture of the calanoid copepod Pseudodiaptomus euryhalinus (Johnson 1939) with different microalgal diets A.C. Puello-Cruz a, S. Mezo-Villalobos a, B. González-Rodríguez a, D. Voltolina b,⁎ a b
Centro de Investigaciones en Alimentación y Desarrollo A.C., Unidad Mazatlán, Sinaloa, Mexico Centro de Investigaciones Biológicas del Noroeste, Laboratorio UAS-CIBNOR, P.O. Box 1132, Mazatlán, Sinaloa, Mexico
a r t i c l e
i n f o
Article history: Received 16 June 2008 Received in revised form 14 February 2009 Accepted 16 February 2009 Keywords: Copepod culture Yield Monoalgal diets Mixed diets
a b s t r a c t The copepod Pseudodiaptomus euryhalinus was fed 320 cells μL− 1 of monoalgal cultures of Chaetoceros muelleri, Nannochloropsis oculata, Isochrysis galbana, Tetraselmis suecica, or a commercial frozen concentrate of Tetraselmis sp., and the diet which gave the best production was compared in a second experiment to three mixed diets: C. muelleri:I. galbana supplied in 1:1 and 2:1 cell ratios and C. muelleri:I. galbana:frozen Tetraselmis sp. in 2:2:1 ratio. These gave better results than the cultures of N. oculata, I. galbana, T. suecica and the frozen Tetraselmis concentrate, but the production was similar to that obtained with C. muelleri supplied as a monoalgal diet, showing that the addition of C. muelleri may improve the performance of other monoalgal diets, whereas the addition of other microalgae is unlikely to improve the results obtained when C. muelleri is supplied as a monoalgal diet. © 2009 Elsevier B.V. All rights reserved.
1. Introduction Several studies have shown that copepods have a higher nutritional value than other live food sources for many fish larvae, because their broad size range, high digestibility and high food value allow better growth and development than other live diets (Støttrup and Norsker, 1997; McKinnon et al., 2003; Rippingale and Payne, 2001). In addition, calanoids are planktonic and have a discontinuous swimming behaviour, which is an important visual stimulus for marine fish larvae (Delbare et al., 1996; Schipp, 2006). However, the techniques for copepod mass culture are at their early stage of development and in particular little is known on the diets that might allow large-scale cultures for sustained and reliable biomass production. Most calanoid copepods are filter-feeders, but there is little information on their natural food preference and on the individual food value of the species present in natural phytoplankton. For this reason, the identification of microalgal diets that allow good survival, fast rates of somatic growth and of reproduction as well as high copepod yields is of paramount importance for larval fish culture. There is a wide range of microalgae available for aquaculture, mostly chosen for their specific nutritional qualities and because they may be easily grown in mass cultures. Some of the most widely
⁎ Corresponding author. Tel.: +52 669 981 03 77; fax: +52 669 982 86 56. E-mail address: [email protected] (D. Voltolina). 0044-8486/$ – see front matter © 2009 Elsevier B.V. All rights reserved. doi:10.1016/j.aquaculture.2009.02.016
used in commercial aquaculture are Chaetoceros muelleri Lemmerman, Isochrysis galbana Parke, Tetraselmis suecica (Kylin) Butcher and Nannochloropsis oculata (Droop) Hibberd (Coutteau, 1996; Duerr et al., 1998). However, monospecific diets may cause nutritional deficiencies, because of the inadequate content of one or more essential nutrients. To reduce this risk, several authors have suggested the use of mixed diets, because their combined nutrient contents are more likely to meet the nutritional requirements of the target species (Brown et al., 1989; Smith et al., 1992). In the case of copepods, this was confirmed by Milione et al. (2007), who found that the development rate of the calanoid Acartia sinjiensis was significantly better with a mixture of two microalgae than with monospecific diets. Semicontinuous cultures of Pseudodiaptomus euryhalinus Johnson fed a mixed algal diet (N. oculata, I. galbana, C. muelleri and T. suecica) have been routinely used in our laboratory since several years to feed the early larval stages of the native puffer fish Sphoeroides annulatus Jenyns and of the red snapper Lutjanus guttatus (Steindachner) (García-Ortega, 2009). The aim of this work was to determine if it was possible to simplify the culture procedure using monoalgal diets, or to improve the productivity of our P. euryhalinus cultures with simpler mixed diets. The feeding rates of filter-feeders may be modified by the external particle concentration or by the volume of the food particles available or ingested (Støttrup and Jensen, 1990; Yúfera and Lubián, 1990), but models based on food availability or carbon ingestion are inadequate to explain copepod egg production (Ohman and Runge, 1994; Pond et al., 1996; Mayor et al., 2006). For this reason, in spite of the large
318
A.C. Puello-Cruz et al. / Aquaculture 290 (2009) 317–319
Table 1 Cell volume (μm3), ash-free dry weight and energy content (AFW in pg cell− 1 and J 10− 6 cell− 1) of the four microalgae used in this study.
a
Chaetoceros Isochrysisa Nannochloropsisa Tetraselmisb a b
μm3
AFW pg cell− 1
J cell− 1
140.2 35.3 8.2 329.4
34.8 17.1 6.3 89.2
0.745 0.496 0.134 1.99
Lora-Vilchis (2004). Piña et al. (2006).
differences in size, individual organic biomass and energy contents of the algal strains, the diets were supplied in equal cell numbers.
2. Materials and methods The 300 L cultures of P. euryhalinus of the Nutrition and Larviculture Laboratory of the Centro de Investigaciones en Alimentación y Desarrollo, Mazatlán Unit, are maintained at 35‰ salinity, 27 ± 1 °C and 12:12 h light:dark photoperiod, food is a 1:1:1:1 mixture in equal cell numbers of N. oculata, I. galbana, C. muelleri and T. suecica supplied daily in the appropriate volumes to maintain a near constant cell concentration of 320 cells μL− 1. Partial harvests (75%) are after two weeks, and the yields obtained with these cultures range between 1.46 and 3.93 organisms mL− 1. The experiments started placing five copulating pairs of P. euryhalinus with no egg sac in 1-L round bottom glass flasks with 500 mL of 5-μm filtered, UV-sterilized seawater (salinity 35 ± 1‰), with gentle aeration and 12:12 h light:dark photoperiod. The flasks were kept in a water bath to maintain temperature at 26 ± 1 °C. The first experiment served to evaluate the monoalgal diets C. muelleri, I. galbana, N. oculata, T. suecica (Table 1), and a commercial frozen concentrate of Tetraselmis sp. (similar in size to T. suecica); in the second experiment, the best treatment of experiment 1 was compared to three mixed diets formulated using C. muelleri, I. galbana and the frozen commercial product. C. muelleri was used as the control treatment, as well as in the mixed diets (1:1 and 2:1 cell number ratio with the second best diet I. galbana). The third was a mixture of C. muelleri + I. galbana + frozen Tetraselmis sp. (2:2:1, respectively). The feeding rates of filter-feeders may be modified by the external particle concentration or by the volume of the food particles available or ingested (Støttrup and Jensen, 1990; Yúfera and Lubián, 1990). However, it has been shown that models based on food availability or carbon ingestion are inadequate to explain copepod egg production (Ohman and Runge, 1994; Pond et al., 1996; Mayor et al., 2006). For this reason, in spite of the large differences in size, individual organic biomass and energy contents of the algal strains (Table 1), the diets were supplied in equal cell numbers. In all cases, each diet was evaluated in five replicate cultures. The algal concentrations were evaluated daily in each culture with a haemacytometer and the appropriate volumes of microalgae culture were added to maintain a stable daily food ration of 320 cells μL− 1.
After addition, the culture volume was corrected to 500 mL. Eighty percent of the seawater was renewed every third day. The experiments lasted 15 days. At the end, all organisms present in each culture were counted, separating ovigerous females, copulating pairs, copepodites and nauplii. After verification of the hypothesis of normality and equal variances, the mean total concentrations were compared using nonparametric or parametric ANOVA tests (first and second experiments) with α = 0.05, separating the different means with the respective Tukey's and Dunn's multiple comparison tests. 3. Results The mean total yield of the cultures fed C. muelleri was significantly higher than those obtained at the end of the first experiment with the other diets. I. galbana gave the second best result followed by the N. oculata and frozen Tetraselmis-fed cultures, which were not significantly different from each other but gave better results than those obtained with T. suecica (Table 2). These results were used to design the mixed diets for the second experiment. C. muelleri was used as the control treatment, as well as in the mixed diets (1:1 and 2:1 cell number ratio with the second best diet I. galbana). The third was a mixture of C. muelleri + I. galbana + frozen Tetraselmis sp. (2:2:1, respectively). The results of this second experiment showed that C. muelleri seems to support practically all the nutritional requirements of P. euryhalinus, since none of the mixtures could improve the nutritional value of monospecific C. muelleri cultures. The mean yields obtained in this experiment ranged from 462.92 ± 152.5 organism L− 1 (C. muelleri) to 296 ± 114.74 L− 1 with the mixture C. muelleri, I. galbana and frozen Tetraselmis sp. (Table 3), and there were no significant differences (P= 0.116) between diets. 4. Discussion The performance of a diet depends on its digestibility, composition and available energy content. Thus, the poor performance of the Nannochloropsis-based diet might be explained by the toughness and poor digestibility of its cell wall (Dhont and Lavens, 1996; Payne and Rippingale, 2000), or by its small size and low weight and energy content (close to 8.2 µm3, 6.3 pg and 0.134 10–6 J cell–1). However, size does not explain the poor results obtained with T. suecica, which are comparable to those found by Cripe (1997) and Piña et al. (2006) with Tetraselmis-fed penaeid shrimp larvae, and to the high degree of deformities and complete lack of development beyond the copepodite stage observed by Knuckey et al. (2005) in Tetraselmis-fed A. sinjiensis nauplii. In this case, digestibility might be the reason for the poor performance of this microalga, in view that production improved when it was supplied as a frozen and probably more digestible diet. Even in this case, this species gave mostly nauplii and copepodites with low adult yields, which could be due to the relatively low food value of this strain, or to the fact that frozen microalgae tend to adhere
Table 2 Production of Pseudodiaptomus euryhalinus with monospecific diets: TE F = Tetraselmis sp. commercial frozen concentrate; TE = Tetraselmis suecica; IS = Isochrysis galbana; CH = Chaetoceros muelleri; NN = Nannochloropsis oculata.
Table 3 Production (organisms L− 1) and stages of development (in %) of Pseudodiaptomus euryhalinus with C. muelleri supplied as a monospecific diet (control) and. with the mixed diets CI 1 = C. muelleri+I. galbana (1:1 ratio), CI 2 = C. muelleri+I. galbana (2:1 ratio) and CIT = C. muelleri+I. galbana + frozen Tetraselmis sp. (2:2:1 ratio).
Diet
TE F
TE
IS
CH
NN
Diet
Control
CI 1
CI 2
CIT
Org. L− 1 CPO CP CD Na
67.7 ± 36.7c – 2% 78% 20%
3 ± 2.9d – 1% 68% 31%
210.4 ± 137.3b 7% 6% 59% 28%
581.6 ± 333.7a 10% 13% 33% 44%
81.7 ± 44.3c 3% 10% 44% 43%
Org. L− 1 CPO CP CD Na
462.9 ± 152.5a 7% 10% 47% 36%
451.6 ± 177.8a 6% 11% 44% 39%
364 ± 167.5a 9% 13% 44% 34%
296 ± 114.7a 13% 17% 38% 32%
Different letters indicate significant difference (one way ANOVA, α = 0.05, a N b N c N d). CPO = copulating adults, ovigerous female; CP = copulating adults; CD = copepodites; Na = nauplii.
The equal letter indicates lack of significant difference (one way ANOVA, α = 0.05). CPO = copulating adults, ovigerous females; CP = copulating adults; CD = copepodites; Na = nauplii.
A.C. Puello-Cruz et al. / Aquaculture 290 (2009) 317–319
to swimming appendages, thus reducing the swimming and feeding activities of the organisms in culture (Paniagua-Chávez and Voltolina, 1995; Isiordia-Pérez and Puello-Cruz, 2007). C. muelleri and I. galbana are considered among the best food sources for filter-feeders, because of their high contents of essential fatty acids that promote high survival and growth (Watanabe et al., 1983; Brown et al., 1989). In the case of copepods, this has been confirmed by the high yields obtained by Støttrup and Jensen (1990) and Payne and Rippingale (2000) with these two microalgae supplied as monospecific diets. However, most authors agree on the high food value of C. muelleri and other single-celled Chaetoceros species for a wide variety of aquatic invertebrates (His and Robert, 1987; Cordero-Esquivel and Voltolina, 1994; Lora-Vilchis et al., 2004b; Suchar and Cigbu, 2006), indicating a better balance of the essential biochemical components of these species. In addition, several aquatic organisms fed with I. galbana have shown poor growth (Lora-Vilchis and Doktor, 2001; Lora-Vilchis et al., 2004a; Piña et al., 2006) and low survival (Helm and Laing, 1987; Cripe, 1997) indicating that, because of species-specific differences in dietary requirements, this species might be a poor source of some essential nutrient. For instance, on a mass to mass basis C. muelleri is a better source of ascorbic acid (Brown and Miller, 1992), as well as of the essential unsaturated fatty acids EPA and ARA (Coutteau, 1996; Piña et al., 2006), whereas I. galbana is richer in DHA. It has been shown that DHA, as well as n−3 fatty acids, may be converted into the essential EPA (Navarro et al., 1999), which may be taken as an indication of a high demand for this nutrient, since there is evidence that it is used as substrate by some copepod species (Veloza et al., 2006). The content of EPA of C. muelleri is close to five times higher than that of I. galbana (Lora-Vilchis et al., 2004a). This would cause a lower demand on the reserves of other fatty acids, which is the likely explanation for the good results we obtained with this diet. Mixed microalgae diets have been suggested for filter-feeders, because monoalgal cultures may not meet their nutritional requirements (Treece, 1984; Brown et al., 1989; Smith et al., 1992; Kleppel and Burkart, 1995). However, this is not a general rule (Yúfera and Lubián, 1990) and it was not the case in this study, because the production we obtained with one of the monospecific cultures, although lower than the 1 copepod mL− 1 mentioned by Payne and Rippingale (2000) for a different copepod species, could not be improved supplying different mixtures with other microalgae species. Acknowledgements Research was funded by the SAGARPA-CONACYT Project 2004C01-196. Gabriela Velasco supplied the microalgae cultures. References Brown, M.R., Miller, K.A., 1992. The ascorbic acid content of eleven species of microalgae used in mariculture. J. Appl. Phycol. 4, 205–215. Brown, M.R., Jeffrey, S.W., Garland, C.D., 1989. Nutritional Aspects of Microalgae Used in Mariculture; a Literature Review. Mar. Lab. Rep., vol. 205. CSIRO. 44 pp. Cordero-Esquivel, B., Voltolina, D., 1994. Growth of Mytilus galloprovincialis Lmk. with four microalgae and two feeding regimes. J. World Aquac. Soc. 25, 471–476. Coutteau, P., 1996. Micro-algae. In: Lavens, P., Sorgeloos, P. (Eds.), Manual on the Production And use of Live Food for Aquaculture. Fish. Tech. Pap., vol. 361. FAO, pp. 7–48. Cripe, G.M., 1997. Spawning and larval survival of the pink shrimp, Penaeus duorarum, in a small culture facility. J. Appl. Aquac. 7, 29–41. Delbare, D., Dhert, P., Lavens, P., 1996. Zooplankton. In: Lavens, P., Sorgeloos, P. (Eds.), Manual on the Production and Use of Live Food for Aquaculture. . Fish. Tech. Pap., vol. 361. FAO, pp. 252–282. Dhont, J., Lavens, P., 1996. Tank production and use of ongrown Artemia. In: Lavens, P., Sorgeloos, P. (Eds.), Manual on the Production and Use of Live Food for Aquaculture. Fish. Tech. Pap., vol. 361. FAO, pp. 164–195. Duerr, E.O., Molnar, A., Sato, V., 1998. Cultured microalgae as aquaculture feeds. J. Mar. Biotechnol. 7, 65–70.
319
García-Ortega, A., 2009. Nutrition and feeding research in the spotted rose snapper (Lutjanus guttatus) and bulleye puffer (Sphoeroides annulatus), new species for marine aquaculture. Fish Physiol. Biochem. 35, 69–80. Helm, M.M., Laing, I., 1987. Preliminary observations on the nutritional value of “Tahiti Isochrysis” to bivalve larvae. Aquaculture 62, 281–288. His, E., Robert, R., 1987. Croissance des larves de Crassostrea gigas et de Mytilus galloprovincialis en présence d'algues monocellulaires isolée du tractus digestif des veligéres du milieu naturel. Haliotis 16, 383–391. Isiordia-Pérez, E., Puello-Cruz, A., 2007. Evaluación del crecimiento y supervivencia en larvas de camarón blanco Litopenaeus vannamei usando como fuente de alimento microalgas vivas y congeladas. REDVET. Rev. Electrón. Vet., vol. 8(5). www. veterinaria.org/revistas/redvet/n070706.html (downloaded July 2008). Kleppel, G.S., Burkart, C.A., 1995. Egg Production and the Nutritional Environment of Acartia tonsa: the Role of Food Quality in Copepod Nutrition. J. Mar. Sci., 52. ICES, pp. 297–304. Knuckey, R.M., Semmens, G.L., Mayer, R.J., Rimmer, M.A., 2005. Development of an optimal microalgal diet for the culture of the calanoid copepod Acartia sinjiensis: effect of algal species and feed concentration on copepod development. Aquaculture 249, 339–351. Lora-Vilchis M.C. 2004. Evaluación de la calidad dietética de microalgas mediante el estudio del balance energético de Artemia franciscana. Ph. D. Thesis. Centro de Investigaciones Biológicas del Noroeste. La Paz, Mexico. 135 pp. Lora-Vilchis, M.C., Doktor, N., 2001. Evaluation of seven algal diets for spat of the pacific scallop Argopecten ventricosus. J. World Aquac. Soc. 32, 228–235. Lora-Vilchis, M.C., Cordero-Esquivel, B., Voltolina, D., 2004a. Growth of Artemia franciscana fed Isochrysis sp. and Chaetoceros muelleri during its early life stages. Aquac. Res. 35, 1086–1091. Lora-Vilchis, M.C., Ruiz-Velasco Cruz, E., Reynoso-Granados, T., Voltolina, D., 2004b. Evaluation of five microalgae diets for juvenile pen shells Atrina maura (Sowerby, 1835). J. World Aquac. Soc. 35, 232–236. Mayor, D.H.J., Anderson, T.R., Irigoien, X., Harris, R., 2006. Feeding and reproduction of Calanus finmarchicus during non-bloom conditions in the Irminger Sea. J. Plankton Res. 28, 1167–1179. McKinnon, A.D., Duggan, S., Nichols, P.D., Rimmer, M.A., Semmens, G., Robino, B., 2003. The potential of tropical paracalanoid copepods as live feeds in aquaculture. Aquaculture 223, 89–106. Milione, M., Zeng, C., Tropical Crustacean Aquaculture Research Group, 2007. The effects of algal diets on population growth and egg hatching success of the tropical calanoid copepod, Acartia sinjiensis. Aquaculture 273, 656–664. Navarro, J.C., Henderson, R.J., McEvoy, L.A., Bell, M.B., Amat, F., 1999. Lipid conversion during enrichment of Artemia. Aquaculture 174, 155–166. Ohman, M.D., Runge, J.A., 1994. Sustained fecundity when phytoplankton resources are in short supply: omnivory by Calanus finmarchicus in the Gulf of St. Lawrence. Limnol. Oceanogr. 39, 21–36. Paniagua-Chávez, C.G., Voltolina, D., 1995. Fresh and frozen Dunaliella sp. (Chlorophyceae, Volvocales) as feed for Artemia franciscana Kellogg (Crustacea, Branchiopoda). Riv. Ital. Acquac. 30, 19–23. Payne, M.F., Rippingale, R.J., 2000. Evaluation of diets for culture of the calanoid copepod Gladioferens imparipes. Aquaculture 187, 85–96. Piña, P., Voltolina, D., Nieves, M., Robles, M., 2006. Survival, development and growth of the Pacific white shrimp Litopenaeus vannamei protozoea larvae, fed monoalgal and mixed diets. Aquaculture 253, 523–530. Pond, D., Harris, R.P., Head, R.N., Harbour, D., 1996. Environmental and nutritional factors determining seasonal variability in the fecundity and egg viability of Calanus helgolandicus in coastal waters off Plymouth, U.K. Mar. Ecol., Prog. Ser. 143, 45–63. Rippingale, R.J., Payne, M.F., 2001. Intensive cultivation of a calanoid copepod Gladioferens imparipes. A Guide to Procedures. Dept of Environmental Biology. Curtin University of Technology. Schipp, G., 2006. The use of calanoid copepods in semi-intensive tropical marine fish larviculture. In: Cruz-Suárez, L.E., Rique-Marie, D., Tapia-Salazar, M., Nieto-López, M.G., Villareal-Cavazos, D.A., Puello-Cruz, A.C., García-Ortega, A. (Eds.), Avances en Nutrición Acuícola VIII. 8th Int. Symp. Aquatic Nutrition. Univ. Autón. Nuevo León, Monterrey. 84–94 pp. Smith, L.L., Biedenbach, J.M., Lawrence, A.L., 1992. Penaeid larviculture: Galveston method. In: Fast, A.E., Lester, L.J. (Eds.), Marine Shrimp Culture, Principles and Practices. Elsevier Science, Amsterdam, pp. 171–191. Støttrup, J.G., Jensen, J., 1990. Influence of algal diet on feeding and egg-production of the calanoid copepod Acartia tonsa Dana. J. Exp. Mar. Biol. Ecol. 141, 87–105. Støttrup, J.G., Norsker, N.H., 1997. Production and use of copepods in marine fish larviculture. Aquaculture 155, 231–247. Suchar, V.A., Chigbu, P., 2006. The effects of algae species and densities on the population growth of the marine rotifer, Colurella dicentra. J. Exp. Mar. Biol. Ecol. 337, 96–102. Treece, G.D., 1984. Larval rearing technology. In: Chamberlain, G.M., Haby, M.G., Miget, R.J. (Eds.), Texas Shrimp Farming Manual: an Update on Current Technology. Texas Agricultural Extension Service, Texas A & M University, Corpus Christi, pp. 43–64. Veloza, A., Chu, F.-L., Tang, K.W., 2006. Trophic modification of essential fatty acids by heterotrophic protists and its effects on the fatty acid composition of the copepod Acartia tonsa. Mar. Biol. 148, 779–788. Watanabe, T., Katajima, C., Fujita, S., 1983. Nutritional value or live organisms used in Japan for mass propagation of fish: a review. Aquaculture 34, 115–143. Yúfera, M., Lubián, L.M., 1990. Effects of microalgal diet on growth and development of invertebrates in marine aquaculture. In: Akatsuka, I. (Ed.), Introduction to Applied Phycology. Academic Publishing, The Hague. 209–227 pp.