Studies on bioaccumulation patterns, biochemical and genotoxic effects of copper on freshwater fish, Catla catla: an in vivo analysis

Abstract During present study, the copper (Cu) mediated oxidative stress was measured that induced DNA damage by concentrating in the tissues of fish, Catla catla (14.45±1.24g; 84.68±1.45mm) (Hamilton,1822). Fish fingerlings were retained in 5 groups for 14, 28, 42, 56, 70 and 84 days of the exposure period. They were treated with 2/3, 1/3, 1/4 and 1/5 (T1-T4) of 96h lethal concentration of copper. Controls were run along with all the treatments for the same durations. A significant (p 0.05) dose and time dependent concentration of Cu was observed in the gills, liver, kidney, muscles, and brain of C. catla. Among organs, the liver showed a significantly higher concentration of Cu followed by gills, kidney, brain, and muscles. Copper accumulation in these organs caused a significant variation in the activities of enzymes viz. superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD). The SOD activity varied significantly in response to the exposure time of Cu as 56 > 70 > 42 > 84 > 28 > 14 days while CAT activity exhibited an inverse relationship with the increase in Cu concentration. POD activity showed a significant rise with an increase in Cu exposure duration. Comet assay exhibited significant DNA damage in the peripheral erythrocytes of Cu exposed C. catla. Among four exposure concentrations, 2/3rd of LC50 (T1) caused significantly higher damage to the nuclei compared to control. Increased POD and SOD activity, as well as a decrease in CAT activity in response to Cu, demonstrates the involvement of a protective mechanism against reactive oxygen species (ROS), whereas increased ROS resulted in higher DNA damage. These above-mentioned molecular markers can be efficiently used for the biomonitoring of aquatic environments and conservation of edible fish fauna.

Resumo Durante o presente estudo, o estresse oxidativo mediado pelo cobre (Cu) foi medido que induziu danos ao DNA por concentração nos tecidos de peixes, Catla catla (14,45 ± 1,24g; 84,68 ± 1,45mm) (Hamilton, 1822). Os alevinos foram retidos em 5 grupos por 14, 28, 42, 56, 70 e 84 dias do período de exposição. Eles foram tratados com 2/3, 1/3, 1/4 e 1/5 (T1-T4) de 96h de concentração letal de cobre. Os controles foram executados junto com todos os tratamentos para as mesmas durações. Uma significativa (p 0,05) concentração dependente do tempo e da dose de Cu foi observada nas brânquias, fígado, rim, músculos e cérebro de C. catla. Entre os órgãos, o fígado apresentou uma concentração significativamente maior de cobre, seguido por guelras, rins, cérebro e músculos. O acúmulo de cobre nesses órgãos causou uma variação significativa nas atividades das enzimas viz. superóxido dismutase (SOD), catalase (CAT) e peroxidase (POD). A atividade de SOD variou significativamente em resposta ao tempo de exposição de Cu como 56> 70> 42> 84> 28> 14 dias, enquanto a atividade de CAT exibiu uma relação inversa com o aumento na concentração de Cu. A atividade POD mostrou um aumento significativo com um aumento na duração da exposição ao Cu. O ensaio do cometa exibiu dano significativo ao DNA induzido por Cu nos eritrócitos periféricos de C. catla. Entre as quatro concentrações de exposição, 2/3 do LC50 (T1) causou danos significativamente maiores aos núcleos em comparação com o controle. O aumento da atividade de POD e SOD, bem como uma diminuição na atividade de CAT em resposta ao Cu, demonstra o envolvimento de um mecanismo protetor contra espécies reativas de oxigênio (ROS), enquanto o aumento de ROS resultou em maior dano ao DNA. Esses marcadores moleculares mencionados acima podem ser usados de forma eficiente para o biomonitoramento de ambientes aquáticos e conservação da ictiofauna comestível.

Studies on bioaccumulation patterns, biochemical and genotoxic effects of copper on freshwater fish, Catla catla: an in vivo analysis / Estudos sobre padrões de bioacumulação, efeitos bioquímicos e genotóxicos do cobre em peixes de água doce, Catla catla: uma análise in vivo

During present study, the copper (Cu) mediated oxidative stress was measured that induced DNA damage by concentrating in the tissues of fish, Catla catla (14.45±1.24g; 84.68±1.45mm) (Hamilton,1822). Fish fingerlings were retained in 5 groups for 14, 28, 42, 56, 70 and 84 days of the exposure period. They were treated with 2/3, 1/3, 1/4 and 1/5 (T1-T4) of 96h lethal concentration of copper. Controls were run along with all the treatments for the same durations. A significant (p < 0.05) dose and time dependent concentration of Cu was observed in the gills, liver, kidney, muscles, and brain of C. catla. Among organs, the liver showed a significantly higher concentration of Cu followed by gills, kidney, brain, and muscles. Copper accumulation in these organs caused a significant variation in the activities of enzymes viz. superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD). The SOD activity varied significantly in response to the exposure time of Cu as 56 > 70 > 42 > 84 > 28 > 14 days while CAT activity exhibited an inverse relationship with the increase in Cu concentration. POD activity showed a significant rise with an increase in Cu exposure duration. Comet assay exhibited significant DNA damage in the peripheral erythrocytes of Cu exposed C. catla. Among four exposure concentrations, 2/3rd of LC50 (T1) caused significantly higher damage to the nuclei compared to control. Increased POD and SOD activity, as well as a decrease in CAT activity in response to Cu, demonstrates the involvement of a protective mechanism against reactive oxygen species (ROS), whereas increased ROS resulted in higher DNA damage. These above-mentioned molecular markers can be efficiently used for the biomonitoring of aquatic environments and conservation of edible fish fauna.

Durante o presente estudo, o estresse oxidativo mediado pelo cobre (Cu) foi medido que induziu danos ao DNA por concentração nos tecidos de peixes, Catla catla (14,45 ± 1,24g; 84,68 ± 1,45mm) (Hamilton, 1822). Os alevinos foram retidos em 5 grupos por 14, 28, 42, 56, 70 e 84 dias do período de exposição. Eles foram tratados com 2/3, 1/3, 1/4 e 1/5 (T1-T4) de 96h de concentração letal de cobre. Os controles foram executados junto com todos os tratamentos para as mesmas durações. Uma significativa (p <0,05) concentração dependente do tempo e da dose de Cu foi observada nas brânquias, fígado, rim, músculos e cérebro de C. catla. Entre os órgãos, o fígado apresentou uma concentração significativamente maior de cobre, seguido por guelras, rins, cérebro e músculos. O acúmulo de cobre nesses órgãos causou uma variação significativa nas atividades das enzimas viz. superóxido dismutase (SOD), catalase (CAT) e peroxidase (POD). A atividade de SOD variou significativamente em resposta ao tempo de exposição de Cu como 56> 70> 42> 84> 28> 14 dias, enquanto a atividade de CAT exibiu uma relação inversa com o aumento na concentração de Cu. A atividade POD mostrou um aumento significativo com um aumento na duração da exposição ao Cu. O ensaio do cometa exibiu dano significativo ao DNA induzido por Cu nos eritrócitos periféricos de C. catla. Entre as quatro concentrações de exposição, 2/3 do LC50 (T1) causou danos significativamente maiores aos núcleos em comparação com o controle. O aumento da atividade de POD e SOD, bem como uma diminuição na atividade de CAT em resposta ao Cu, demonstra o envolvimento de um mecanismo protetor contra espécies reativas de oxigênio (ROS), enquanto o aumento de ROS resultou em maior dano ao DNA. Esses marcadores moleculares mencionados acima podem ser usados ​​de forma eficiente para o biomonitoramento de ambientes aquáticos e conservação da ictiofauna comestível.

Induced breeding of Catla catla carried out at low temperature in FRP carp hatchery of Arunachal Pradesh, India

Aim: The present study was undertaken to find out the possibilities of induced breeding of Catla catla, at low water temperature using fibre reinforced (FRP) tanks in a carp hatchery. Methodology: Brood fish of C. catla was maintained in 0.1 ha pond with proper feeding and pond management. Six induced breeding programmes: 3 in April (pre-monsoon) and 3 in June (monsoon) of 2017 were conducted with C. catla using synthetic hormone, Ovasis intra-peritoneally. The relationship between temperature variation and latency period, effective spawning period, fecundity, hatching start time, hatching duration and spawn production with its recovery was observed. Results: The latency period (time gap between injection and first egg release) was very high and ranged between 653.3 and 721.6 min for breeding in the pre-monsoon and ranged between 446.6 and 480 min for monsoon period. In pre-monsoon, the effective spawning period between 86.6 and 116.6 min and in monsoon between 53.3 and 73.3 min; both varied significantly (p˂0.05). In pre-monsoon period, the average water temperature during larval incubation in hatching pool ranged between 21.58 and 22.58ºC, whereas during monsoon period it ranged between 30.1 and 30.4ºC. The spawn production was 0.13 to 0.22; and 0.47 to 0.65 (lakh kg-1 b. wt. of female) in pre-monsoon and monsoon period, respectively, and showed a significant difference (p˂0.05). Interpretation: This report on breeding of C. catla in the temperature range of 18.6-24.7oC indicates the possibility of spawn production of Indian Major Carp in the north east hilly region states of India

Histological studies on ontogenic development of lymphoid organs in Indian major carp, Catla catla

Aim: Immunostimulation, before the fish is able to mount an effective immune response, can induce tolerance. Therefore, present study was carried out to investigate the structural development of lymphoid organs, namely thymus, kidney and spleen of Indian major carp, Catla (Catla catla), which will help in deciding the appropriate age, when immunocompetence is established. Methodology: In this study, ontogeny of the lymphoid organs of catla was studied from hatching up to 54 days post hatch (dph). For this, whole fish were fixed, processed and embedded in paraffin wax. Further, serial sections were cut, stained and observed under microscope. Results: The thymus anlage appeared in the dorso-anterior part of the gill cavity at 1 dph. At 5 dph, an undifferentiated mass of stem cells was observed and by 8 dph, it appeared lymphoid. At 11 dph, clear dark and light zones of cells were observed. At 26 dph, Hassall’s corpuscles were observed and no further changes were observed except increase in number of lymphoid cells till end of the sampling period. Kidney was observed as an undifferentiated mass of stem cells at 1 dph with few renal tubules. At 5 dph, undifferentiated stem cells increased in number, and by 8 dph, few erythropoietic cells were observed. At 11 dph, lymphoid cells were present and by 17 dph, renal tubules started degenerating. At 26 dph, anterior kidney completely lacked excretory tissue and appeared lymphoid. Spleen was observed at 3 dph as an undifferentiated mass of stem cells, and at 8 dph, erythrocytes were seen. At 11 dph, spleen was heavily packed with erythrocytes, whereas at 20 dph, the number of erythrocytes decreased and lymphocytes were observed. Fully developed spleen was observed at 33 dph with presence of red and white pulp.

Physiological responses of Catla catla larvae fed with Achyranthes aspera seed enriched diet and exposed to UV-B radiation.

The antioxidant and growth stimulating properties of seeds of Achyranthes aspera were evaluated on UV-B irradiated Catla catla (catla) larvae. Catla larvae (initial weight: 1.2 ± 0.01 mg) were fed with four different diets — D1, D2 and D3 containing 0.1, 0.25 and 0.5% seeds of A. aspera and D4, control diet for 35 days. The larvae were then exposed to UV-B radiation (80 µW/cm2) on every alternate day for 20 days. Survival, growth, tissue glutamic oxaloacetic transminase (GOT), tissue glutamate pyruvate transminase (GPT), thiobarbituric acid reactive substances (TBARS) and nitric oxide synthase (NOS) were studied in larvae on day-21 of irradiation. Significantly (P < 0.05) higher survival and average weight were found in D3 diet fed fish compared to the other groups. Survival rate was 8-16% higher in seed enriched diet fed groups, compared to the control one. Higher levels of GOT and GPT found in control diet fed larvae showed the degree of tissue damage due to UV-B exposure. Significantly (P < 0.05) lower level of GPT in D3 indicated the UV-B protective effect of the seed of A. Aspera (earlier, the presence of ecdysterone, essential fatty acids and amino acids, polyphenolic compounds, steroids, etc. has been reported from seed). TBARS which indicated the level of tissue lipid peroxidation were significantly (P < 0.05) higher in control group, compared to the other feeding schemes. NOS level was significantly (P < 0.05) higher in D2 and D3, compared to the D1 and control groups. In conclusion, supplementation of A. aspera seed (0.5%) improved the physiological condition (in terms of reduce lipid oxidation and better immune system) and gave bioprotection to catla larvae challenged with UV-B stress.

Immune response of catla catla fed with an oral immunostimulant plumbago rosea and postchallenged with aeromonas hydrophila.

The present study was designed to evaluate the immunostimulant potential of Plumbago rosea in fish catla, post challenged with Aeromonas hydrophila. Fishes were divided into 2 groups one received control diet and another received Plumbago rosea incorporated diet for 14 days. The haematological parameters and serum protein were estimated between control and experiment. There is An increase in TEC (P<0.05) and TLC (P<0.05) in the immunostimulant administered (IS)diet was observed. Increase in lymphocyte count was noted in immunostimulant incorporated diet. Remarkable increase in Hb from 6.2 to 8g% and serum protein level from 0.4to 0.5g% was observed in experimental fishes The immunostimulant administered Catla catla when challenged with Aeromonas hydrophila showed a decrease in TEC and an increase in TLC. Increase in lymphocytes was also noted.

Studies on the immouostimulant activity of Coriandrum sativum and resistance to Aeromonas hydrophila in Catla catla.

The present study was designed to evaluate the immunostimulant potential of Coriandrum sativum is fish Catla catla, post challenged with A.hydrophila. Fish were divided into 2 groups one received control diet, another received Coriandrum Sativum incorporated diet for 14 days. The haematological parameters and serum protein were analyzed between control and experiment. There was a significant increase in TEC (P<0.01) and TLC (P <0.01) in the immunostimulant (IS) administered diet. Remarkable increase in Hb content from as 6.9.to 7.39% and serum protein level from 0.56 to 0.58g% was noted. The immunostimulant administered post challenged Catla catla showed a decrease in TEC at 103 cfu/ml and a drastic increase in TLC(<0.01) in both 103 and 105 cfu/ml Hb content on IS incorporated post challenged Catla catla showed an increase from 8.7 – 11.5g%.

Genetic variation of wild and hatchery populations of the catla Indian major carp (Catla catla Hamilton 1822: Cypriniformes, Cyprinidae) revealed by RAPD markers

Genetic variation is a key component for improving a stock through selective breeding programs. Randomly amplified polymorphic DNA (RAPD) markers were used to assess genetic variation in three wild population of the catla carp (Catla catla Hamilton 1822) in the Halda, Jamuna and Padma rivers and one hatchery population in Bangladesh. Five decamer random primers were used to amplify RAPD markers from 30 fish from each population. Thirty of the 55 scorable bands were polymorphic, indicating some degree of genetic variation in all the populations. The proportion of polymorphic loci and gene diversity values reflected a relatively higher level of genetic variation in the Halda population. Sixteen of the 30 polymorphic loci showed a significant (p < 0.05, p < 0.01, p < 0.001) departure from homogeneity and the F ST values in the different populations indicated some degree of genetic differentiation in the population pairs. Estimated genetic distances between populations were directly correlated with geographical distances. The unweighted pair group method with averages (UPGMA) dendrogram showed two clusters, the Halda population forming one cluster and the other populations the second cluster. Genetic variation of C. catla is a useful trait for developing a good management strategy for maintaining genetic quality of the species.