Effects of anesthesia with the essential oil of Ocimum gratissimum L. in parameters of fish stress / Efeitos da anestesia com o óleo essencial de Ocimum gratissimum L. em parâmetros de estresse de peixes

RESUMO: The effects of anesthesia with the essential oil of Ocimum gratissimum (EOO) in parameters of stress after handling were investigated in silver catfish (Rhamdia quelen). EOO was obtained from the aerial parts by hydrodistillation. Juveniles were anesthetized with 70 or 300 mg L-1 EOO and submitted to air exposure for 1 minute. The fishes were sampled immediately or transferred to anesthetic-free aquaria until sampling. In the first experiment, juveniles had their blood collected at 0, 1, 4, and 8 h after handling to assay plasma cortisol and blood glucose levels. The unanesthetized animals were restrained manually for blood collection. In the second experiment, water samples of the recovery aquaria were collected to evaluate net ion fluxes at 0 - 4 h and 4 - 8 h. Water and ethanol controls were also performed under the same conditions. The results showed that the cortisol levels did not differ among the treatments. Hyperglycemia was verified in fish exposed to 70 and 300 mg L-1 EOO at 1 h and 4 h after handling. After 8 h, cortisol and glucose concentrations were lower or similar than those from immediately after handling for all treatments. EOO anesthesia prevented Na+ efflux observed in the control groups in both flux periods. There were net Cl- and K+ effluxes at 0 - 4 h and influxes at 4 - 8 h after handling in most treatments, and these fluxes did not differ among the treatments. The results suggest that EOO did not impair stress recovery and did not act as an additional handling stressor in silver catfish.

RESUMO: Os efeitos da anestesia com o óleo essencial de Ocimum gratissimum (EOO) em parâmetros de estresse após manuseio foram investigados em jundiás (Rhamdia quelen). EOO foi obtido a partir das partes aéreas por hidrodestilação. Os juvenis foram anestesiados com 70 ou 300 mg L-1 de EOO e expostos ao ar por 1 minuto. Os peixes foram amostrados imediatamente ou transferidos para aquários sem anestésico até amostragem. No primeiro experimento, os juvenis tiveram seu sangue coletado em 0, 1, 4, e 8 h após manuseio para avaliar os níveis de cortisol e glicemia. Os animais não anestesiados foram contidos manualmente para coleta sanguínea. No segundo experimento, amostras de água foram coletadas do aquário de recuperação dos animais para avaliação do fluxo iônico entre 0 - 4 h e 4 - 8 h. Grupos controles em água e etanol também foram realizados sobre as mesmas condições. Os resultados demonstraram que os níveis de cortisol não diferiram entre os tratamentos. Hiperglicemia foi detectada em peixes expostos a 70 e 300 mg L-1 de EOO em 1 h e 4 h após o manuseio. Após 8 h, os teores de cortisol e glicose foram menores ou similares aqueles imediatamente após o manuseio para todos os tratamentos. A anestesia com EOO preveniu o efluxo de Na+ observado para os grupos controle em ambos os períodos avaliados. Ocorreram efluxos de Cl- and K+ entre 0 - 4 h e influxos entre 4 - 8 h após o manuseio para a maioria dos tratamentos, e estes eventos não diferiram entre os tratamentos. Os resultados sugerem que o EOO não prejudica a recuperação do animal frente ao evento estressor ou atua como estressor adicional ao manuseio em jundiás.

Increased immunoglobulin production in silver catfish (Rhamdia quelen) exposed to agrichemicals.

Fish vaccination has been increasingly exploited as a tool to control pathogen infection. The production of immunoglobulin following vaccination might be affected by several factors such as management procedures, water temperature, and the presence of xenobiotics. In the present study, we aimed to investigate the kinetics of immunoglobulin production in silver catfish (Rhamdia quelen) inoculated with inactivated Aeromonas hydrophila and kept at two different water temperatures (17.4±0.46 or 21.3±0.36C). The effect of a second antigen inoculation and exposure of fish to sublethal concentrations of the herbicides atrazine and glyphosate at 10% of the lethal concentration (LC50-96h) on specific serum antibodies were also investigated. Antibodies to A. hydrophila were detected as early as 7 days post-inoculation and increased steadily up to 35 days. The kinetics of antibody production were similar in fish kept at 17.4±0.46 and 21.3±0.36C, and reinoculation of antigen at 21 days after priming failed to increase specific antibody levels. Intriguingly, we found that, in fish exposed to atrazine and glyphosate, the secretion of specific antibodies was higher than in non-exposed inoculated fish. These findings are important for the design of vaccines and vaccination strategies in Neotropical fish species. However, because atrazine and glyphosate are widespread contaminants of soil and water, their immune-stimulating effect could be harmful, in that fish living in herbicide-contaminated water might have increased concentrations of nonspecific antibodies that could mediate tissue injury.

Stress in Salminus brasiliensis fingerlings due to different densities and times of transportation / Estresse em alevinos de dourado (Salminus brasiliensis) em diferentes densidades e tempos de transporte

The stress in dorado fingerlings (Salminus brasiliensis) caused by transportation at densities of 5, 10, and 15g/l after 4, 8, and 12h was evaluated by the concentration of tissue cortisol measured by ELISA. The conditions of transportation were simulated on an orbital table shaker with horizontal movements, inside 15 litres plastic bags filled with 4 litres of water and pure oxygen. Cortisol concentrations increased in all densities after 4h of transportation converging to a common concentration at the end of the tested times. Electrical conductivity of water increased with density and transportation time. The transportation caused stress on fish, but the increase on density and in times of transportation did not cause mortality to fingerlings. The transportation of S. brasiliensis fingerlings can be done without mortality or apparent injuries to animals until the maximum analyzed density of 15g/l and up to 12h.

O estresse em alevinos de dourado (Salminus brasiliensis) causado pelo transporte nas densidades 5, 10 e 15g/l após 4, 8 e 12h, foi avaliado por meio dos níveis de cortisol tecidual quantificado por ELISA. O transporte foi simulado em uma mesa agitadora com movimento orbital, em sacos plásticos de 15 litros, contendo 4 litros de água e oxigênio puro. As concentrações de cortisol aumentaram em todas as densidades após 4h de transporte e convergiram para uma mesma concentração ao final dos tempos de transporte. A condutividade elétrica da água aumentou com a densidade e com os tempos de transporte. O transporte causou estresse aos peixes, mas o aumento da densidade e do tempo de transporte não causou mortalidade nos alevinos. O transporte de alevinos de S. brasiliensis pode ser realizado até a densidade máxima avaliada de 15g/l e o tempo máximo de 12h sem que haja mortalidade ou danos aparentes aos animais.

Embryonic and larval development of Jundiá (Rhamdia quelen, Quoy & Gaimard, 1824, Pisces, Teleostei), a South American Catfish

The jundiá (Rhamdia quelen, Quoy & Gaimard) is an endemic South American fish species. Because this species supports cold winters and grows faster during warm months, it has begun to be viewed as an ideal species for fish production in southern South America. In the present study, jundiá oocytes used were obtained by extrusion from females after hormone injection. Soon after hydration, the eggs were transferred to 50 L conic glass incubators, with constant and controlled water influx. Samples of fertilized eggs were transferred to Petri dishes and, examined under a stereoscopic microscope, were spherical, demersal, and non-adhesive with defined perivitelline space and resistant chorion. Cleavage stages occurred during the first 3.5 h. After hatching, larvae were transferred to 200 L glass fiber incubators. First signs of embryo movement were observed 21 h after fertilization; larval eclosion occurred 30.5 h after fertilization. Present findings may provide a basis for studies aimed at determining the complete ontogeny of jundiá and may be useful in eco-toxicological studies.

O jundiá (Rhamdia quelen, Quoy & Gaimard) é uma espécie endêmica da América do Sul. Por ser adaptada ao frio do inverno e ter um crescimento rápido durante os meses quentes, o jundiá é uma espécie adequada para aqüicultura no sul da América do Sul. Muitos aspectos da fisiologia reprodutiva, larvicultura, hematologia, fisiologia da resposta ao estresse, têm sido recentemente estudados. Os ovócitos utilizados neste estudo foram obtidos pela extrusão de fêmeas após indução hormonal. Logo após a hidratação, foram transferidos para incubadoras cônicas de vidro com capacidade para 50 L, com fluxo de água constante e controlado. Amostras de ovos fertilizados foram colocadas em placas de Petri e examinadas através de estereomicroscópio. Os ovos eram esféricos, demersais e não-adesivos, com espaço perivitelino definido e córion resistente. Os estágios de clivagem ocorreram durante as 3,5 primeiras horas. Após a eclosão, as larvas foram transferidas para incubadoras de fibra de vidro de 200 l. Os primeiros sinais de movimento embrionário foram observados 21 h após a fertilização, e a eclosão das larvas ocorreu 30,5 h após a fertilização. Estes resultados podem servir como base para muitos estudos, objetivando o conhecimento da ontogenia completa do jundiá, e para aplicação em estudos ecotoxicológicos.

Embryonic and larval development of jundiá (Rhamdia quelen, Quoy and Gaimard, 1824, Pisces, TeleosteI), a South American catfish.

The jundiá (Rhamdia quelen, Quoy and Gaimard) is an endemic South American fish species. Because this species supports cold winters and grows faster during warm months, it has begun to be viewed as an ideal species for fish production in southern South America. In the present study, jundiá oocytes used were obtained by extrusion from females after hormone injection. Soon after hydration, the eggs were transferred to 50 L conic glass incubators, with constant and controlled water influx. Samples of fertilized eggs were transferred to Petri dishes and, examined under a stereoscopic microscope, were spherical, demersal, and non-adhesive with defined perivitelline space and resistant chorion. Cleavage stages occurred during the first 3.5 h. After hatching, larvae were transferred to 200 L glass fiber incubators. First signs of embryo movement were observed 21 h after fertilization; larval eclosion occurred 30.5 h after fertilization. Present findings may provide a basis for studies aimed at determining the complete ontogeny of jundiá and may be useful in eco-toxicological studies.