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The Effect of Salinity on Osmoregulation and Development of the Juvenile Fat Snook, Centropomus Parallelus (POEY)

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The Effect of Salinity on Osmoregulation and Development of the Juvenile Fat Snook, Centropomus Parallelus (POEY) The effect of salinity on osmoregulation and development of the juvenile fat snook, Centropomus parallelus (POEY) Sterzelecki, FC.a*, Rodrigues, E.b, Fanta, E.a and Ribeiro, CAO.a aSetor de Ciências Biológicas, Departamento de Biologia Celular, Universidade Federal do Paraná, Centro Politécnico, CEP 81531-990, Curitiba, PR, Brazil bDepartamento de Bioquímica, Universidade de Taubaté, CEP 12030-180, Taubaté, SP, Brazil *e-mail: [email protected] Received: February 3, 2012 – Accepted: September 3, 2012 – Distributed: August 31, 2013 (With 2 figures) Abstract Eurihaline fish support waters with different salt concentration. However, numerous studies have shown that salinity can affect fish development. Thus, the effect of salinity change from 20 to 5 and 35 on survival, weight, length, gill chloride cell ultrastructure and gill Na+,K+ ATPase activity was evaluated in Centropomus parallelus following short-term (6, 24 and 96 hours) and long-term exposure (30 and 60 days). Salinity did not affect C. parallelus survival, final weight and length. The quantity of chloride cells increased visibly at salinities of 5 and 35, with the cells exhibit- ing the typical features of uptake and secretory cells, respectively. Na+,K+ ATPase activity in the gill of the C. parallelus was significantly greater at a salinity of 5 than at a salinity of 20 or 35 after 96 hours, but not after 30 or 60 days. These results indicate that salinity change from high to low salt water induces gill chloride cell and Na+,K+ ATPase activity adaptations after short-term exposure. However, after long-term exposure at salinity 5, gill Na+,K+ ATPase activity is no more necessary at high levels. The increase in salinity to 35 does not induce significant change in gills. Juveniles of C. parallelus may thus be capable of acclimating to salinities of 5 to 35 for 60 days without signifi- cant effects on development. Keywords: Centropomus parallelus, chloride cell, osmoregulation, Na+,K+ ATPase. O efeito da salinidade sobre a osmoregulação e o desenvolvimento do juvenil de robalo-peva, Centropomus parallelus (POEY) Resumo Peixes eurihalinos suportam águas com diferentes concentrações de sal. Contudo, muitos estudos têm mostrado que a salinidade pode afetar o desenvolvimento do peixe. Portanto, o efeito da mudança de salinidade de 20 para5e35na taxa de sobrevivência, peso, comprimento, morfologia das células de cloreto branquiais e atividade da Na+,K+ ATPase foram avaliadas no Centropomus parallelus após curto (6, 24 e 96 horas) e longo tempo de exposição (30 e 60 dias). A salinidade não afetou a sobrevivência, o peso e comprimento final do robalo-peva. A quantidade de células de cloreto aumentou visivelmente nas salinidades 5 e 35, exibindo morfologias típicas de células que absorvem e secretam sal, respectivamente. A atividade da Na+,K+ ATPase nas brânquias do C. parallelus foi significativamente maior na salinidade 5 do que nas salinidades 20 ou 35 após 96 horas, mas nãoapós 30 e 60 dias. Esses resultados indicam que a mudança de alta para baixa salinidade provoca adaptações nas células de cloreto e na atividade da Na+,K+ ATPase branquial em curto prazo. Contudo, após longa exposição na salinidade 5, a alta atividade da Na+,K+ ATPase branquial não é mais necessária. O aumento de salinidade para 35 não induz mudanças significativas nas brânquias. Portanto, juvenis de C. parallelus possuem a capacidade de aclimatação nas salinidades de5a35semefeitos significativos no desenvolvimento após 60 dias. Palavras-chave: Centropomus parallelus, célula de cloreto, osmoregulação, Na+,K+ ATPase. satisfactory growth rate and suitability for human con- 1. Introduction sumption. Although technology for the mass production The fat snook, Centropomus parallelus, is important of juvenile C. parallelus is currently available, Cerqueira for commercial and recreational fisheries as well as for and Tsuzuki (2009) appointed that further research is aquaculture. This fish has been considered to be a good needed to reach the cost-effective growth-out technol- candidate for cultivation due to its resistance to diseases, ogy. Braz. J. Biol., 2013, vol. 73, no. 3, p. 609-615 609 Sterzelecki, FC. et al. The fat snook is a species of the genus Centropomus confirmed using a refractometer (± 1). Over an acclima- that lives in and around estuarine waters located in south- tion period of nine days, low levels of total-ammonia- eastern Florida to the southern coast of Brazil (Rivas, nitrogen (TAN), nitrite and nitrate (0.3 to 0.6 mg/L, 1986). Although juveniles of C. parallelus are naturally 0.1 mg/L and 5-10 mg/L, respectively) were maintained found in a wide salinity environment, it has been shown using a recirculation system with biological and mechan- that salinity does affect fish development (Boeuf and ical filters. Dissolved oxygen was maintained between Payan, 2001). In previous experiments, salinity specifi- 4-5 mg/L and pH at 8-8.2. The snooks were kept at cally affected C. parallelus food intake, growth (Rocha et 26 ± 2 °C , under natural photoperiod and fed once a day al., 2005) and digestive enzyme activity (Tsuzuki et al., to satiation with adult of frozen Artemia. After feeding, 2007b). Nevertheless, the relationship between the os- all waste at the bottom of the tank was siphoned. moregulatory system and development at different salini- Following the acclimation period, the salinity was ties has yet to be explored in C. parallelus. Several progressively increased and decreased in two aquariums studies have associated the rapid growth of some species to 35 and 5, respectively. The third group was cultivated with a reduction in osmoregulatory costs (Boeuf and at a salinity of 20, although the same volume of water was Payan, 2001; Sampaio and Bianchini, 2002; Laiz-Car- exchanged as in the other groups to maintain consistency. rión et al., 2005). Additional chemical properties of the water were main- The environmental salinity different from the inter- tained at levels similar to those achieved during the accli- nal osmotic concentration of teleost fish provokes loss or mation period by monitoring with Sera® Kit and gain of salt and water. To maintain the ionic concentra- changing weekly 20% of water. tion of body fluids, the osmoregulatory organs work to- gether (McCormick, 2001). The gills are probably the 2.2. Fish measurements and sample collection organs that consume the most energy during osmo- From experiment I, 54 to 58 fish in each aquarium regulation (Sangiao-Alvarellos et al., 2003). More spe- started the trial. After acute (6, 24 and 96 hours) and + + cifically, the gill Na ,K ATPase, located mainly in chronic period (30 and 60 days) of salinity change, 3 to chloride cells, requires energy input and plays an impor- 10 fish were sampled. From the experiment II, 51 to 57 tant role in osmoregulation in both hyperosmotic fish in each aquarium started the trial and 2 to 10 fish (Jobling, 1995; Evans et al., 2005) and hyposmotic (Ev- were sampled at the same time point as before. For both ans, 2008; Parks et al., 2008) environments. The activity experiments, sampled fish were anesthetised with benzo- of this ATPase spends at least half of the energy con- caine (50 mg/L), weighed, measured and sacrificed to + + sumed during Na /H exchange in ionocytes (Kirschner, evaluate Na+,K+ ATPase activity and gill chloride cells. + + 2004). For this reason, Na ,K ATPase is considered to After 96(t0) hours, 30 (t30) and 60 (t60) days all the fish be a good biomarker of osmoregulation in teleosts. Un- were measured and weighed to biometric comparison derstanding the activity of this pump at different salt con- (see Table I). The sampling variation occurred due to the centrations may help to clarify the appropriate salinity for sacrifice to biochemical analysis, mortality and cannibal- farm juvenile fat snooks. The goal of the present study ism. was to assess the relationship between Na+,K+ ATPase activity, chloride cell ultrastructure, survival, final 2.3. Na+,K+ ATPase activity weight and length in C. parallelus following acclimation The branchial arches from sacrificed fish, with the to different salinities. exception of the second arch, were frozen immediately in liquid nitrogen and stored at -80 °C until the biochemical 2. Materials and Methods assays were performed. A method obtained from McCormick (1993) was used to evaluate Na+,K+ ATPase 2.1. Experimental design activity. The gills were homogenised in 100 mLofSEI The experiment was conducted in February (experi- buffer (pH 7.5; 250 mM sucrose; 10 mM Na2EDTA and ment I) and repeated under the same conditions in May, 50 mM imidazole) and 50 mL of SEID (0.1 g sodium 2007 (experiment II). A total of 328 juvenile of C. deoxycholic acid in 33.3 mL of SEI). Following ho- parallelus were obtained from the Marine Fish Culture mogenisation, the gills were centrifuged at 5,000 g for Laboratory (LAPMAR) at the Universidade Federal de 2 min. The insoluble material was separated, and 10 mL Santa Catarina, Brazil. Both trial fish derived from the of supernatant in duplicate was added into microplate same broodstock, but from a different spawning time. wells to 200 mL of the reaction mixture containing They were studied 65 days after hatching, weighing 50 mM imidazole (pH 7.5), 189 mM NaCl, 10.5 mM 0.14 ± 0.05 g and 0.13 ± 0.04 g (mean ± SD) in experi- MgCl2, 2.86 U/mL LDH, 3.57 U/mL PK, 2 mM PEP, ment I and experiment II, respectively. They were trans- 150 mM NADH and 0.5 mM ATP. In addition, other two ported to the Universidade Federal do Paraná and sepa- wells of microplate were filled with the same mixture rated into three 100-L glass aquariums filled with water plus 0.5 mM ouabain. NADH oxidation levels at 28 °C at a salinity of 20, at a stocking density of 0.6 fish/L.
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