Preliminary Assessment of Growth of FlatfishParalichthys adspersus Juveniles in Different Culture Systems Lili Carrera, Noemí Cota, Angélica Castro, Melissa Montes and Jorge Tam

FIGURE 1. Juvenile culture of flatfishParalichthys adspersus in a static system.

In Peru, aquaculture sector Peruvian cuisine. Additionally, policy is driven by the National in recent years there has been a Aquaculture Development decrease in the volume of Plan 2010-2021 (DS N ° fisheries catches. Some years 001-2010-PRODUCE) that ago flatfish reproduction and guides research to strengthen culture were developed under the value chain of consolidated captive conditions in public and emerging species, and and private institutions, which promote and execute aquaculture conducted experimental research diversification with native and production at a commercial species that have market pilot level to achieve sustained potential and can become production of the species. the basis for major industries. However, there continues to be The National Program problems of quality, quantity and for Aquaculture Science, growth of juveniles to support Technological Development grow-out operations. and Innovation 2013-2021 (RP FIGURE 2. Juvenile culture of flatfish adspersus in a recirculating The Peruvian Marine No. 064-2013-CONCYTEC-P) system at the IMARPE laboratory. Research Institute (IMARPE) established priorities for has been conducting research on aquaculture research and development. One research line is oriented reproduction, larval development and juvenile rearing of P. adspersus to developing a culture technique for the flatfishParalichthys in the laboratory. In addition, and with great effort, Pacific Deep adspersus and transferring the technology to the commercial sector. Frozen SA (PDFSA), a commercial producer and marketer of quality The focus is on improving the techniques for reproduction, larval hydrobiological products has also initiated the culture of marine development and production of quality fingerlings. fish in its aquaculture facility in Huarmey (Ancash, Peru), including Paralichthys adspersus is distributed from Paita, Peru, to commercial production of flatfish. Lota and Juan Fernández Islands in Chile (Acuña and Cid 1995, In this context, the objective of the study reported here was to Chirichigno 1998, Sielfeld et al. 2003). It is a species of commercial compare the effect of different culture systems on growth of juvenile importance for the national market because it is a valuable fish in P. adspersus.

44 SEPTEMBER 2017 • WORLD AQUACULTURE • WWW.WAS.ORG FIGURE 3. Biometric sampling of juvenile Paralichthys adspersus in a recirculating system at the IMARPE laboratory.

FIGURE 4. Juvenile culture of Paralichthys adspersus in recirculating system FIGURE 5. Biometric sampling of Paralichthys adspersus in a recirculating at commercial pilot level at Pacific Deep Frozen S.A. aquaculture system at commercial pilot level at Pacific Deep Frozen S.A.

Broodstock Management The management and protocol of larval and post-larval feeding are The Fish Culture Laboratory of the Alexander von Humboldt summarized in Table 1. Aquaculture Research Center of IMARPE has a stock of P. adspersus At PDFSA, larvae were also grown in static systems in 1000-L breeders in two recirculating aquaculture systems (RAS) (Carrera fiberglass tanks, with seawater (35 UPS) filtered and sterilized by UV et al. 2013). Gonadal maturation was induced through application radiation. Photoperiod was controlled with an automatic timer and set of a regular photoperiod (12 light hours and 12 dark hours) and a to 12 h light and 12 h dark. The management and protocol of larval and temperature of 16.7 ± 0.8 C. For spawning, a mature female was post-larval feeding are summarized in Table 2. selected with an average oocyte diameter > 500 μm and two males with sperm motility greater than 50 percent. A synthetic analogue of Juvenile Culture gonadotropin-releasing hormone (buserelin acetate, Conceptase®) Juveniles were reared in three types of systems: 1) a static system was given to the female by intraperitoneal injection at 0.1 mL/kg (SS) from 110 to 194 DPH (~ 1 kg/m2), 2) a RAS in the Fish Culture and, in the case of the two males, 0.1 mL/kg fish by intramuscular Laboratory of IMARPE (RAS-L) from 103 to 194 DPH (~ 2 kg/ injection. Broodstock management and spawning technique carried m2) and 3) a RAS at a commercial pilot level in the Culture Center out at PDFSA was similar to that at IMARPE, also in RAS. of PDFSA (RAS-C) from 105 to 196 DPH, where juveniles were maintained at a higher density (~ 4 kg/m2). In all systems, water filtered Larval Development through 1 μm mesh and sterilized by UV radiation was used. At IMARPE, larvae were cultured in static systems from 1 day At IMARPE, in the SS, juveniles were held in eight fiberglass post hatch (DPH) in 150-L fiberglass tanks with seawater (35 UPS) tanks of 500-L capacity (Fig. 1). A daily water exchange of 80 to 100 filtered and sterilized by ultraviolet (UV) radiation, with constant percent was applied and biometric samples were collected every fifteen aeration and illumination between 905 and 1245 lux. Continuous light days. In the RAS-L, individuals were kept in two 700-L fiberglass was applied for the first ten days of culture and then a photoperiod of tanks and the components of the system included a biofilter, UV light 12 h light and 12 h dark was applied until the end of the experiment. (CONTINUED ON PAGE 46)

WWW.WAS.ORG • WORLD AQUACULTURE • SEPTEMBER 2017 45 TABLE 1. Larval and post-larval culture protocol of the flatfishParalichthys adspersus under laboratory conditions at IMARPE.

DPH Water FEEDING Bottom Cleaning exchange(%) Microalgae Rotifers Artemia sp. Microdiet siphoning of the tank Larval Phase 1 - 5 0 x x 6 50 x x 7 - 10 0 x x 11 10 x x 12 - 14 15 x x 15 - 20 20 x x x x 21 - 23 50 x x x x x 24 - 27 100 x x x x x 28 - 40 100 x x x Post-larval Phase 40 - 50 100 x x x x 50 - 60 100 x x x

TABLE 2. Larval and post-larval culture in Paralichthys adspersus at PDFSA.

DPH Water FEEDING Bottom Cleaning exchange(%) Microalgae Rotifers Artemia sp. Microdiet siphoning of the tank Larval Phase 1-3 0 x x 4-10 10 x x 11-14 25 x x 15 25 x x 16-19 30 x x 20-23 40 x x x x 24-26 50 x x x x x 27 70 x x x x 28-35 100 x x x x Post-larval Phase 35-50 100 x x x x 50-60 100 x x x

water sterilizer, heat pump and water pump (Fig. 2). Sampling was C and was relatively stable in all systems, with the greatest variation performed monthly (Fig. 3) and the daily water exchange rate was in the static system (Fig. 6). Average temperatures were 20.5 ± 1.0 C, 10-20 percent. 21.1 ± 1.1 C and 19.8 ± 0.7 C in SS, RAS-L and RAS-C, respectively. Feed was supplied to apparent satiety or ad libitum with extruded Thiese temperatures are greater than those (14.9-17.3 C) used by pellets1 from 0.84 to 2.1 mm particle size. Feed was provided four Silva (2001), whose reached the commercial size of 1 kg times per day, between 2 and 4 percent of total biomass. In addition, in 3.5 years. Wasielesky et al. (1998) observed that juveniles of P. physicochemical parameters were monitored daily. orbignyanus accepted feed at a temperature range of 10-27 C, with an At PDFSA, commercial juvenile culture was carried out in optimum temperature range in the pre-fattening period of 18-21 C. It is six fiberglass tanks of 1500 L coupled to two RAS (Fig. 4). These important to determine species-specific optimal temperature ranges at systems consisted of a sedimentation tank, sand filter, biological filter, each stage of the life cycle. degasser and UV sterilizer. For maintenance of the system, water Mean values of dissolved oxygen concentration were 7.7 ± 0.6 was replaced at 15 to 20 percent/d, with biweekly sampling during mg/L, 6.2 ± 0.6 mg/L and 5.7 ± 1.1 mg/L in SS, RASL and RAS-C, the culture period (Fig. 5). Extruded pellets of 1-2 mm particle size respectively. Average values of pH were 7.7 ± 0.3 and 7.3 ± 0.7, in the (Otohime1, Aquaxcel®2 and Nicovita3) was supplied ad libitum three SS and RAS-L, respectively; this parameter was not measured in the to six times, depending on consumption. RAS-C. Water temperature in all systems was maintained between 19-22 In all culture systems, total length (cm) and weight (g) were

46 SEPTEMBER 2017 • WORLD AQUACULTURE • WWW.WAS.ORG measured to obtain the specific growth al. 2001) and open flow in Senegalese sole rate, condition factor (Innis 1990), (Sánchez et al. 2010). The present research coefficient of variation, calculated evaluated the growth of P. adspersus juve- according to Merino et al. (2007). niles in different culture systems, obtaining a greater growth in recirculating systems Results and Discussion than in a static system. Silva and Oliva The final total length and weight of (2010) obtained an average size of 8.9 cm juveniles in the two recirculating systems and a weight and 9.5 g in juvenile culture was similar to each other and greater than of P. adspersus. Growth of P. adspersus that in the static system (Table 3). The juveniles was less than that of , which specific growth rate of juvenile flatfish reached 9-10 g in 90-100 DPH (Stoss et al. cultured in the RAS-L was greater than 2004). that cultured in RAS-C or SS (Table 3). An important culture variable is den- Other values for specific growth rate (on sity, which is inversely related to growth weight) reported for this species and other FIGURE 6. Water temperature in different culture rate (Ashley 2007, Kiessling et al. 2007) in species of flounder include P. adspersus systems used to culture juvenile Paralichthys adspersus. flatfish species such as Solea solea (Howell 1.7 percent/d 1998, Schram et (Silva and Oliva al. 2006), Scoph- 2010), P. or- thalmus maximus bignyanus 1.8 (Irwin et al. 1999) percent/d (Sam- and P. californicus paio et al. 2001) (Merino et al. and Solea spp. 2007), among oth- 2.1 percent/d ers. In the research (Reig 2001). The reported here, length-weight greater growth exponent of fish rate was observed in all systems in the RAS, where was around 3, fish were cultured indicating iso- at a higher den- metric growth sity (~ 2-4 kg/m2), (Table 3). than in the static In recent FIGURE 7. Growth in length (a) and weight (b) of juvenileParalichthys adspersus in different culture systems. culture system (~ years, flatfish 1 kg/m2). culture has been developing in Peru. Carrera et al. (2013) reported the Density has a direct effect on growth variability in flatfish such as conditioning of broodstock in recirculating aquaculture systems. In turbot, where a more pronounced increase in variation of body weight the second stage of the project, juvenile culture was carried out at an was found at higher densities (Irwin et al. 1999). The coefficient of experimental level at IMARPE laboratories and as a commercial pilot variation on fish size was least in the RAS-L than in the other two at PDFSA. systems. In the RAS-L, frequent size grading was carried out to Different systems have been used to culture flatfish juveniles, such reduce size heterogeneity in culture tanks. as recirculating aquaculture systems in California flounder (Conklin There were small differences in condition factor of fish cultured et al. 2003), semi-recirculation in the Brazilian flounder (Sampaio et in the three systems (Table 3). The condition factor was similar to (CONTINUED ON PAGE 48)

TABLE 3. Biometric information of Paralichthys adspersus juveniles cultured in various culture systems.

SYSTEM Parameter Static RAS-L RAS-C Final total length (cm) 7.7 ± 0.2 9.9 ± 0.2 10.5 ± 0.1 Final weight (g) 6.5 ± 0.5 11.4 ± 0.6 15.6 ± 0.6 SGR length (%/d) 0.7 ± 0.1 0.9 ± 0.2 0.7 ± 0.1 SGR weight (%/d) 1.8 ± 0.4 2.7 ± 0.5 2.0 ± 0.2 K 1.33 ± 0.10 1.26 ± 0.01 1.16 ± 0.03 CV weight (%) 40.7 ± 4.1 29.1 ± 1.6 39.4 ± 1.7 b exponent 2.78 3.08 2.94

WWW.WAS.ORG • WORLD AQUACULTURE • SEPTEMBER 2017 47 that of juvenile P. californicus (Merino et al. 2007), with K between Applied Aquaculture 14(3-4):143-154. 1.06 and 1.10, and a lower K at higher densities. The higher values ​​of FAO (Food and Agriculture Organization of the United Nations). 2016. condition factor in the static system could be related to the lowest crop El estado mundial de la pesca y la acuicultura 2016. Contribución a density used in this system (~ 1 kg/m2). A similar inverse relationship la seguridad alimentaria y la nutrición para todos. Rome, Italy between K and crop density was observed by Hachero-Cruzado et al. Hachero-Cruzado, I., M. Herrera, A. Amo and M. Cordero. 2010. (2010) in Scopththalmus rhombus, where a K of 1.56 was obtained at Influence of stocking density on tissue lipid composition in flatfish 1 kg/m2 and a K of 1.47 at 15 kg/m2. Scophthalmus rhombus. Aquaculture Europe Abstract Book The research reported here will serve as the basis for future 10:555-556. research about optimal growing conditions for P. adspersus. Howell, B. 1998. The effect of stocking density on growth and size variation in cultured turbot, Scophthalmus maximus, and sole, Solea Acknowledgment solea. International Council for the Exploration of the Sea CM The research reported here was financed by the National Innovation 1998/L: 10. Program for Competitiveness and Productivity INNÓVATE-PERÚ Innis, D. 1990. Juvenil California halibut, Paralichthys californicus, in the development of Project Agreement No. 236-FINCyT-IA 2013 growth in relation to thermal effluent. Fish Bulletin 174:153-165. “Production of seed of the Paralichthys adspersus in captivity: II Irwin, S., J. O’Halloran and R. FitzGerald. 1999. Stocking density, Improvement of the techniques of larviculture.” Also, appreciation growth and growth variation in juvenile turbot, Scophthalmus is extended to Pacific Deep Frozen S.A. for being part of the project, maximus (Rafinesque). Aquaculture 178(1):77-88. with the participation of engineers Jaime Pauro and Juan Gutiérrez. Kiessling, A., H. Yavuzcan, B. Damsgard, A. Roque, J. Turnbull, N. Ducan, S. Kafri and H. Van de Vis. 2007. Cost action 867-Wellfish: Notes Welfare of fish in European aquaculture. CD Abstracts. Proceedings Lili Carrera, Noemí Cota, Angélica Castro, Melissa Montes, of the Aquaculture Europe Congress 24-27:609-610. Laboratorio de Cultivo de Peces, Dirección General de Merino G., R. Piedrahita and D. Conklin. 2007. The effect of fish Investigaciones en Acuicultura; E-mail: [email protected] stocking density on the growth of California halibut (Paralichthys Jorge Tam, Laboratorio de Modelado Oceanográfico, Ecosistémico californicus) juveniles. Aquaculture 267:176-186. y del Cambio Climático, Dirección General de Oceanográfica y Reig, L. 2001. Utilización de un aroma comercial en pienso para Cambio Climático, Instituto del Mar del Perú, Callao 1, Callao, lenguado: Evaluación de su eficacia en el comportamiento y en Perú. el crecimiento. Tesis para optar al Título de Doctor en Xarxa. 1 Otohime ™ brand (Marubeni Nisshin Feed CO., LTD. Sakura Universitat Politecnica de Catalunya. Departament d’Enginyeria Muromachi Building 4F, 4-5-1 Nihonbashimuromachi, Chuo-ku, Hidraulica, Maritima i Ambiental. Tokyo, 103-0022 - Japan) Sampaio, L.A., A. Bianchini and V.R. Cerqueira. 2001. Growth of 2 Aquaexcel® brand (Agribrands Purina Perú S. A., Panamericana juvenile Brazilian flounder, Paralichthys orbignyanus, cultured at Norte Km. 17.5 - Independencia Lima, Perú). different salinities. Journal of Applied Aquaculture 11(1-2):67-75. 3 Nicovita brand (Alicorp S. A. A., Av. Argentina N° 4793 - Callao, Sánchez, P., P.P. Ambrosio and R. Flos. 2010. Stocking density and sex Perú). influence individual growth of Senegalese sole Solea( senegalensis). Aquaculture 300:93-101. References Schram, E., J.W. Van der Heul, A. Kamstra and M.C.J. Verdegem. Acuña, E. and L. Cid. 1995. On the ecology of two sympatric 2006. Stocking density-dependent growth of Dover sole (Solea flounder of the genus Paralichthys in the Bay of Coquimbo, Chile. solea). Aquaculture 252(2):339-347. Netherlands Journal of Sea Research 34(1-2):1-11. Siefeld, W., M. Vargas and I. Kong. 2003. Primer registro de Etropus Ashley, P. 2007. Fish welfare: Current issues in aquaculture. Applied edenes (Jordán 1889), Bothus constellatus (Jordán y Goss Behaviour Science 104:199-235. 1889), Achirus klunzingeri (Steindachner 1880) y Symphurus Carrera, L., N. Cota, M. Montes, E. Mateo, V. Sierralta, T. Castro, A. elongatus (Gunther 1868) (Piscis, Pleuronectiformes) en Chile, Perea, C. Santos and C. Espinoza. 2013. Broodstock management con comentarios sobre la distribución de los lenguados chilenos. of the fine flounder Paralichthys adspersus (Steindachner, 1867) Investigaciones Marinas Valparaíso 31:51-65. using recirculating aquaculture systems. Latin American Journal of Silva, A. 2001. Advances in the culture research of small-eye flounder, Aquatic Research 41(1):89. Paralichthys microps, and Chilean flounder, P. adspersus, in Chile. Chirichigno, N. 1998. Clave para identificar los peces marinos del Journal of Applied Aquaculture 11:147-164. Perú. Instituto del Mar del Perú, Publicación Especial. Callao. 496 Silva, A. and M. Oliva. 2010. Revisión sobre aspectos biológicos y pp. de cultivo del lenguado chileno (Paralichthys adspersus). Latin Comisión de la Pesca Continental y la Acuicultura en América American Journal of Aquatic Research 38(3):377-386. Latina y el Caribe (COPESCAALC). 2016. Panorama de la Pesca Stoss, J., K. Hamre and H. Ottera. 2004. Weaning and nursery. Pages Continental y la Acuicultura en América Latina y el Caribe, 337-362 In: E. Moksness, E. Kjorsvik and Y. Olsen, editors. Culture Décima Cuarta Reunión. Lima. 11 pp. of Coldwater Marine Fish. Blackwell Publishing, Des Moines, IA Conklin, D.E., R.H. Piedrahita, G.E. Merino, J.B. Muguet, D.E. Wasielesky, Jr., W., A. Bianchini and K. Miranda Filho. 1998. Bush, E. Gisbert and M. Cervantes-Trujano. 2003. Development Tolerancia a la temperatura de juveniles de lenguado Paralichthys of California halibut, Paralichthys californicus, culture. Journal of orbignyanus. Frente Marítimo 17 (Sec. A): 55-60.

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