Oxidative stress and histopathologic biomarkers of exposure to bisphenol-A in the freshwater fish, Ctenopharyngodon idella

ABSTRACT Bisphenol-A (BPA) belongs to the family of endocrine disrupting chemicals (EDCs) and it is used in the production of polycarbonate plastic and epoxy resins. The reproductive toxicity of BPA is well documented but it also exerts its toxic effects through multiple pathways especially by inducing a state of oxidative stress and causing damage to the vital organs. In the present study, histopathologic and oxidative damage caused by BPA in liver and kidneys of fresh water cyprinid, Ctenopharyngodon idella was evaluated. LC50 of BPA for Ctenopharyngodon idella was determined by probit regression analysis. Fish were exposed to a sublethal concentration of BPA i.e. 3.2 ppm (1/2 LC50) for 14 days. Histologic studies revealed that BPA caused degenerative changes in liver and kidneys and exposure of sublethal concentration of BPA caused oxidative damage in both organs. Lipid peroxidation significantly increased in liver and kidneys of treated group. Catalase activity and reduced glutathione content significantly decreased in the group exposed to BPA compared to control and glutathione-S-transferase activity increased significantly in both organs exposed to the sublethal concentration of BPA. From this study it is concluded that BPA caused toxic effects in fish species by changing oxidative balance and damaging the vital organs.

Study on comparative histology of retinas in Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans, Gekko japonicus, and Columba livia / Estudio histológico comparado de la retina en Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans, Gekko japonicus, y Columba livia

The objective of this study was to explore the relationship between the retinal structure and its life adaptation to the environment of Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans, Gekko japonicus and Columba livia . Measuring retinal thickness of each layer, the nuclei layer, and the diameter of each nuclear layer of the five animals, the statistical data analysis shows that: the nuclei layers of five animals are all 4, and their structures can be divided to 10 layers when observing with optical microscope. The retinal thickness of Ctenopharyngodon idella was 190.49 mm, Cynops orientalis was 173.07 µm, and the Bufo bufo gargarizans was 195.06 µm, Gekko japonicus was 224.32 µm and Columba livia was 174.10 µm. The number of retinal inner nuclear layers of Bufo bufo gargarizans and Gekko japonicus and Columba livia are more than their outer nuclear layers, on the contrary, retinal inner nuclear layers of Ctenopharyngodon idella and Cynops orientalis are less than their outer nuclear layers. The rod and cone layer of retina of Cynops orientalis were more advanced, but their nerve fiber layer (NFL) degraded highly, revealing a strong photosensitivity but a low visual sensitivity; to Columba livia , their NFL of retina are highly developed, so as their vision. The different structures and functions of the retina of Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans, Gekko japonicus and Columba livia correspond with their behavioral characteristics and the living environment's change from aquatic to amphibious to land.

El objetivo de este estudio fue explorar la relación entre las estructuras de la retina y su adaptación al medioambiente en Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans,Gekko japonicus y Columba livia . La medición del espesor de cada capa de la retina, la capa nuclear y su diámetro en los cinco animales, mostró a través del análisis estadístico que las capas nucleares en todos ellos fueron 4, y sus estructuras se pueden dividir en 10 capas cuando se observan con el microscopio óptico. El espesor de la retina de Ctenopharyngodon idella fue 190,49 µm, de Cynops orientalis fue 173,07 µm, de Bufo bufo gargarizans fue 195,06 µm, de Gekko japonicus fue 224,32 µm y de Columba livia fue 174,10 µm. El número de capas nucleares internas de la retina de Bufo gargarizans, Gekko japonicus y Columba livia fue mayor que sus capas nucleares externas, mientras que las capas nucleares internas de Ctenopharyngodon idella y Cynops orientalis fueron menos que las capas nucleares externas. La capa de conos y bastones de la retina de Cynops orientalis fue más desarrollada, pero su capa de fibras nerviosas presentó una elevada degeneración, lo que muestra una gran fotosensibilidad, pero una sensibilidad visual baja. En Columba livia, la capa de fibras nerviosas de la retina estuvo muy desarrollada, y de esta manera, su visión. El grado de desarrollo de las diferentes estructuras y funciones de la retina de Ctenopharyngodon idella, Cynops orientalis, Bufo bufo gargarizans, Gekko japonicus y Columba livia está relacionada con sus características de comportamiento y el cambio de las condiciones de las vidas acuática y anfibia en la tierra.

Purification and characterization of 6-phosphogluconate dehydrogenase (6-PGD) from grass carp (Ctenopharyngodon idella) hepatopancreas.

6-Phosphogluconate dehydrogenase (6-PGD, E.C.: 1.1.1.44) was purified and characterized from the hepatopancreas of grass carp (Ctenopharyngodon idella) for the first time. Grass carp represents the second largest aquaculture industry in the world after silver carp, constituting 14.7% of the world aquaculture production, with an average annual increase of 14% in China, mainly as a source of food. The purification procedure involved a single 2’, 5’-ADP-Sepharose 4B affinity chromatographic step by using different elution buffers. The enzyme was purified 309-fold with a specific activity of 5.259 U/mg protein and yield of 68%. The purity and subunit molecular weights of the 6-PGD were checked on SDS-PAGE and purified enzyme showed a single band on the gel. The subunit molecular mass was 57 kDa, with an optimum pH, temperature and ionic strength at 7.96, 50oC and 100 mM Tris-HCl, respectively. The Km values of 6-PGA and NADP+ were 0.019 and 0.0052 mM, respectively, while Vm of 6-PGA and NADP+ was 0.69 U/ml. Dissociation constants (Ki) for 6-PGA and NADP+ were 2.05 and 0.12 mM, respectively. NADPH inhibited the enzyme in a competitive manner and its Ki value was 0.032 mM. The Cu2+, Zn2+, Cd2+ and Al3+ showed inhibitory effects on the enzyme with IC50 values of 0.293, 0.099, 0.045 and 1.526 mM, respectively. All tested metals inhibited the enzyme in a competitive manner, indicating that these metals might be toxic even at low concentrations for the 6-PGD. As the fish is one of valuable foodstuff of animal sources for human consumption, under certain environmental conditions, metal ions accumulated in fish up to a lethal concentration may be harmful for human health. Therefore, it is impending to reduce the concentration of metal ions in contaminated lakes and rivers for fishery and also for human health.

Teores de proteína bruta em dietas práticas para juvenis de carpa capim / Crude protein levels in practical diets for grass carp juveniles

Avaliou-se a resposta de juvenis de carpa capim (Ctenopharyngodon idella) à inclusão de proteína bruta (PB) em dietas práticas, em um experimento com 120 juvenis e 80 dias de duração. As dietas eram compostas por farinha de carne suína, farelo de soja, milho e farelo de trigo com 22, 30, 36 ou 44 por cento de PB. Dez juvenis (153,0±1,5g) por tanque (850L) foram alimentados duas vezes ao dia (3 por cento da biomassa). Foi observado efeito linear positivo da PB para peso final, ganho em peso, taxa de crescimento específico, rendimento de carcaça e filé, coeficiente de retenção proteica, taxa de eficiência proteica, deposição de gordura e proteína e hematócrito. O mesmo efeito também foi observado para triglicerídeos e colesterol total no sangue, indicando que a proteína da dieta foi utilizada como fonte energética. Conversão alimentar aparente e gordura no peixe inteiro apresentaram efeito quadrático, com pontos de máxima em 40,6 e 37,1 por cento de PB, respectivamente. Pode-se concluir que a porcentagem mínima de PB para a carpa capim na fase de recria foi de 44 por cento e que a variação da proteína da dieta promoveu alterações no metabolismo dos juvenis de carpa capim, refletidos nos parâmetros sanguíneos e de carcaça.

The response of grass carp (Ctenopharyngodon idella) juveniles to crude protein (CP) levels in practical diets was evaluated in an experiment with 120 juveniles and 80 days of feeding. The diets containing porcine meat meal, soybean meal, corn and wheat meal, with 22, 30, 36, or 44 percent CP. Ten juveniles (153.0±1.5 g) per tank (850L) were fed twice daily with pelleted feed (3 percent of biomass). Linear effect of CP on final weight, weight gain, specific growth rate, carcass and fillet yield, coefficient of protein retention, protein efficiency rate, deposition of fat and protein, and hematocrit were observed. It was also observed effect on triglycerides and total cholesterol in blood, indicating that dietary protein was being used as energy source. Feed conversion ratio and fat content in whole fish showed quadratic effect, with maximum points at 40.6 and 37.1 percent CP, respectively. The best CP content for grass carp in growing phase was 44 percent, and the variation of protein content in diet changed the metabolism of grass carp juveniles, reflected in blood and carcass parameters.

Acute toxicity of nuvan®, an organophosphate to freshwater fish Ctenopharyngodon idella and its effect on oxygen consumption.

The acute toxicity of Nuvan to the grass carp Ctenopharyngodon idella was determined using static and continuous flow through system for 24, 48, 72 and 96 hr. The median lethal concentration (LC50) values were 13.1, 10.9, 9.8 and 6.5 mg l-1 respectively in static system and 10.7, 9.5, 8.0 and 7.5 mg l-1 respectively in continuous flow through system. A reduction in oxygen consumption is observed when the fish is exposed to the toxicant and the mortality is due to effect of metabolism of energy synthesis.