Integrated histopathological lesion (IHL) analysis - a histopathological tool for the evaluation of UV-B exposure on the gill, liver and intestine of Catla catla.

The present study aimed to analyse the integrated histopathological lesions (IHLs) of the gill, liver and intestine of Catla catla exposed to the different doses of UV-B radiation. Gill exhibited the lesions like hypertrophy, hyperplasia, vacuolation, fusion of the gill filaments, rupture in the gill lamellae, epithelial cell lifting and necrosis. The UV-B-exposed liver of Catla showed the lesions like the degeneration of nucleus, the disarrangement of hepatocytes, sinusoidal vacuolation, epitheliod histiocyst, hepatocellular adenoma, exocrine adenoma, cyst formation and diffused epithelial necrosis (DEN). UV-B-exposed intestine showed the lesions like the distortion of columnar epithelial cells (CECs), distortion in lamina propria (LP), disruption in brush border (BB), vacuolation in LP, the presence of submucosal mass (SM), the degeneration of nucleus, the presence of tactoid bodies, the presence of aschoff nodules and metatypical cell carcinoma. These histopathological alterations can be considered as the main blocking alterations of the growth and absorption as well as the final production of fish which can cause a serious loss in total yield to fish farmers which can interrupt the profitable economical production of fish.

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.

First report of Dactylogyrus formosus from India: a case of co-existence with Paradactylogyrus catlaius causing mass mortality in Indian major carp, Catla catla.

The family Dactylogyridae, known for its role as a frequent pathogen in Cyprinids, was identified in a case of mass mortality involving Catla catla fingerlings (measuring 6.5 ± 2.0 cm and weighing 40.5 ± 10 g) in West Bengal, India. Microscopic examination revealed the presence of two co-existing parasites, Dactylogyrus formosus and Paradactylogyrus catlaius, attached to different sections of the gill filament. Despite their coexistence, these parasites exhibited marked differences in their haptoral hard parts, genital organs, and preferred habitats. Molecular analysis of the Internal Transcribed Spacer 1 and 4 genes indicated more than 90% similarity between the detected parasite and D. formosus previously reported in China. Histopathological observations illustrated the parasites' specific attachment to the distal end of the primary gill lamellae, gradually causing destruction to a maximum number of secondary lamellae. Internally, infiltration of eosinophilic granular cells was observed in gill and kidney blood vessels, while the liver exhibited hepatocytes filled with hemosiderin. The infected fish were treated for 24 h with a safe dose of common salt (5.6 ppt) and albendazole (62 ppm). The survivability rate was significantly higher (p < 0.05) in both treated groups compared to the control. Interestingly, the group treated with common salt showed superior results compared to the albendazole-treated fish. This study presents the sympatric speciation of D. formosus in a new host (C. catla) and explores its host specificity, histopathology, and treatment methods. This case marks the first report of D. formosus causing substantial mortality in cultured Catla in India, alongside the coexistence with Paradactylogyrus catlaius.

Influence of photoperiod variations on the mRNA expression pattern of melatonin bio-synthesizing enzyme genes in the pineal organ and retina: A study in relation to the serum melatonin profile in the tropical carp Catla catla.

Surface-dwelling C. catla were exposed to different photoperiods (8L:16D, 12L:12D, 12D:12L and 16L:8D) and the mRNA level profile of enzymes involved in melatonin synthesis was evaluated in the pineal gland and retina. Furthermore, a comparative analysis of the serum melatonin profile with the mRNA level was also performed. The results indicated diurnal variations in the transcripts of tph1, aanat and hiomt in the pineal organ and retina, and these variations change with the change in lighting regime. The serum melatonin profile showed rhythmicity in the natural photoperiod, but the serum melatonin level increased proportionally with increasing daylength. In short photoperiods, the peak value (though lower than in long photoperiods) of melatonin maintains a longer duration in serum. Moreover, the comparative analysis revealed a similar profile of mRNA of pineal aanat1 and aanat2 with serum melatonin under the same lighting conditions. This indicates that serum melatonin is produced by the pineal gland. Our results specify the importance of day length and the timing of onset or offset of the dark for maintaining the oscillating levels of serum melatonin and mRNA levels of melatonin biosynthesizing enzyme genes in the pineal organ and retina as well. The findings in this study highlight the distinctive pattern of mRNA levels in the pineal organ and retina under different photoperiods. The pineal melatonin biosynthesizing enzyme genes showed a similar pattern with serum melatonin levels while the retinal genes changed dramatically with photoperiod. We also revealed a light-dependent transcriptional regulation of pineal aanat genes in C. catla. Moreover, our results suggest that ALAN and skyglow can influence the levels of serum melatonin and its biosynthesis, resulting in desynchronization of the entire biological clock as well as the overall physiology of the animal.

Effects of Illicium verum Hook. f. (Chinese herb) enriched diet on growth performance, immune response and disease resistance in Catla catla [Hamilton] fingerlings against Aeromonashydrophila.

Our present study investigates the effects of Illicium verum enriched diet on growth performance, immune response and disease resistance in Catla catla fingerlings against Aeromonas hydrophila. Experimental groups, i.e., infected C. catla were fed with 5 g/kg, 10 g/kg and 15 g/kg of I. verum diet for a period of 30 days showed a significant increase in their final body weight (FBW) over control. There was a significant increase (p < 0.05) in total weight gain (TWG), specific growth rate (SGR), feed conversion rate (FCR), protein efficiency ratio (PER), condition factor (CF) and survival rate (SR) in 10 g/kg and 15 g/kg groups compared to control; also from 31 to 60 days these groups showed an enhanced (p < 0.05) complement activity, phagocytic activity, respiratory burst activity and lysosome activity. The experimental group fed with 10 g/kg of I. verum diet showed the least cumulative mortality rate (5%) compared to 5 g/kg (30%) and 15 g/kg (15%) experimental groups and in control (10%) and negative control (75%). Our present study highlights that challenged fish fed with the mid dosage of 10 g/kg showed a significant increase in overall growth performance, non susceptibility to diseases and tolerance against A. hydrophila in C. catla compared to other experimental groups including the group given with high dosage of 15 g/kg. Thus our results suggest 10 g/kg as the optimal dosage of I. verum that can be used to enrich the fish diet for a better performance and effective immunostimulation in commercially important C. catla for its sustainable culture and is reported for the first time.

In vivo genotoxic effects of commercial grade cypermethrin on fish peripheral erythrocytes.

The study explicates the genotoxic effects of commercial grade cypermethrin on peripheral erythrocytes of Catla catla, chronically exposed to two environmentally relevant concentrations. The fish was treated with sub-lethal concentrations 0.12 µg/L and 0.41 µg/L (1/10th and /1/3rd of 96 h LC50 ) of cypermethrin for 45 days. DNA damage in the exposed fish was assessed using alkaline comet assay, presence of micronuclei (MN), erythrocyte nuclear and cytoplasmic abnormalities. Exposure to cypermethrin induced a dose-dependent increase in percent DNA damage, micronucleus frequency and erythrocyte abnormalities. Nuclear anomalies such as notched nuclei, lobed nuclei, bridged nuclei, and deformed nuclei; and cytoplasmic anomalies like anisochromasia, vacuolated cytoplasm, lobed cells, and echinocytes were observed. The findings revealed the genotoxic potential of commercial formulations pyrethroid cypermethrin at concentrations found in the environment and their potential deleterious effects on nontarget aquatic organisms.

A hepatic antimicrobial peptide, hepcidin from Indian major carp, Catla catla: molecular identification and functional characterization.

BACKGROUND: Increase of antibiotic resistance in pathogenic microbes necessitated novel molecules for curing infection. Antimicrobial peptides (AMPs) are the gene-encoded evolutionarily conserved small molecules with therapeutic value. AMPs are considered as an alternative drug for conventional antibiotics. Hepcidin, the cysteine-rich antimicrobial peptide, is an important component in innate immune response. In this study, we identified and characterized hepcidin gene from the fish, Catla catla (Indian major carp) and termed it as Cc-Hep. RESULTS: Open reading frame of Cc-Hep consists of 261 base pair that encodes 87 amino acids. Cc-Hep is synthesized as a prepropeptide consisting of 24 amino acid signal peptide, 36 amino acid propeptide, and 26 amino acid mature peptide. Sequence analysis revealed that Cc-Hep shared sequence similarity with hepcidin from Sorsogona tuberculata. Phylogenetic analysis indicated that Cc-Hep was grouped with HAMP2 family. Structure analysis of mature Cc-Hep identified two antiparallel beta sheets stabilized by four disulphide bonds and a random coil. The mature peptide region of Cc-Hep has a charge of + 2, isoelectric value 8.23 and molecular weight 2.73 kDa. CONCLUSION: Functional characterization predicted antibacterial, antioxidant, and anticancer potential of Cc-Hep, which can be explored in aquaculture or human health care.

Changes in immune genes expression, immune response, digestive enzymes -antioxidant status, and growth of catla (Catla catla) fed with Astragalus polysaccharides against edwardsiellosis disease.

The effect of four level of Astragalus polysaccharides (APs) supplementation diets, (CD: control diet and three experiment diet (E), EA: 100 mg kg-1 APs; EB: 200 mg kg-1 APs; EC: 300 mg kg-1 APs) on growth, changes in haemato-biochemical parameters and metabolic-digestive enzymes, enhancement of antioxidant activity, innate-adaptive immune response, and cytokine gene expression were studied in catla (Catla catla) against Edwardsiella tarda. The healthy and challenged groups fed the CD displayed no mortality, while fish fed EA or EC revealed 10% mortality, but the mortality was only 5% in diet EB. Fish fed diet EB and EC revealed significantly better growth rates and high RBC count during the experimental period. Albumin and globulin levels were significant improved when fish were fed the diet EB and EC from weeks 6-8. The superoxide dismutase (SOD) was significant ameliorated by EB feeding from weeks 4-8. In contrast, serum myeloperoxidase (MPO), catalase (CAT), malondialdehyde (MDA)/lipid peroxidation (LPO), glutathione peroxidase (GPx), respiratory burst activity (RBA), bactericidal action (BCA), serum lysozyme activity (SLA), nitric oxide synthase (NOS), head kidney leukocytes response proliferation (HKLP), hemolytic action (HLA), hydrogen peroxides (H2O2), and immunoglobulin (Ig) were significantly improved from week 6-8. Groups fed the APs enriched diets had significant ameliorated interleukin (IL)-1ß and interferon (IFN)-γ mRNA expression after 6 and 8 weeks of feeding. However, IL-10 and major histocompatibility complex (MHC)-1 mRNA expressions were significant enhanced in catla fed all APs diets on week 8. APs enriched diets revealed significant improved tumor necrosis factor (TNF)-α and TNF receptor-associated factor-6 (TRAF6) mRNA expression on week 4, but toll-like receptor-2 (TLR2) and TLR4 mRNA expression were significant enhanced by diet EB and EC after weeks 6 and 8. Similarly, the lysozyme (Lyz)-C and Lyz-G mRNA levels in the head kidney (HK) increased by APs feeding on weeks 6 and 8, whereas the EB diet, the expression of nucleotide binding oligomerization domain-1 (NOD1) was significantly improved on weeks 6 and 8, but NOD2 mRNA expression was only significant enhanced after 8 weeks of diet EB. By feeding healthy catla and E. tarda challenged fish fed diet EB, resulted in significantly increased growth, haemato-biochemical indices, metabolic-digestive enzymes, antioxidant activities, innate-adaptive immune responses, and cytokine gene expression mainly between 6 and 8 weeks.

Dietary thiamin requirement of fingerling major carp Catla catla (Hamilton).

Dietary thiamin requirement of fingerling Catla catla (3.5 ± 0.15 g) was evaluated by feeding casein-gelatin-based iso-nitrogenous (350 g/kg crude protein) and iso-caloric (16.72 kJ/g GE) diets containing six graded levels of thiamin (0, 0.2, 0.4, 0.8, 1.6 and 3.2 mg/kg dry diet) for 12 weeks. Significantly (p < 0.05) higher weight gain (AWG), best feed conversion ratio (FCR), protein retention efficiency (PRE), RNA/DNA ratio and haematological indices were recorded in fish fed diet containing 0.8 mg/kg thiamin. Dietary thiamin supplementation improved transketolase activity (TKA) and maximum value was recorded in fish fed 0.8 mg/kg thiamin beyond which stagnation in TKA activity was evident. Liver thiamin concentration was found to be maximum in fish fed diet containing 1.6 mg/kg thiamin. A significant (p < 0.05) consistent reduction in the hepatic thiobarbituric acid reactive substances (TBARS) activity was displayed with incremental concentration of thiamin up to 0.8 mg/kg, beyond which a reverse trend was evident. However, a significant (p < 0.05) improvement was noted in superoxide dismutase (SOD) and catalase (CAT) activity with the increasing level of dietary thiamin from 0 to 0.8 mg/kg. Broken-line regression analysis of AWG, FCR, PRE and TKA estimated the requirement in the range of 0.74-0.79 mg/kg dry diet.

Purification and characterization of bioactive compounds extracted from Suaeda maritima leaf and its impact on pathogenicity of Pseudomonas aeruginosa in Catla catla fingerlings.

Incidence of various dreadful microbial infections and the development of antibiotic resistance by infection causative microbes are the main reasons for reducing aquaculture productivity. Hence, there is an immense need for the discovery of alternative and efficient treatment for quick recovery of diseased fishes. In the present study, Suaeda maritima leaf extracts (hexane, diethyl ether, ethanol, and water) were screened for in vitro and in vivo antibacterial and antioxidant activities. Out of all the four extracts, ethanolic extract showed highest antibacterial activity against S. aureus (4.9 ± 1.3 mm), B. subtilis (1.6 ± 0.3 mm), K. pneumoniae (4.2 ± 1.8 mm), and P. aeruginosa (4.1 ± 1.2 mm). Similarly, antioxidant activity was also higher for ethanolic extract (500 µg/mL) based on DPPH radical scavenging ability (71.6 ± 1.4%) and reducing potential (149 µg/mL) assays. Further, ethanolic extract was purified consecutively via column chromatography and preparative TLC where an active fraction was selected based on highest antibacterial (10.1 ± 1.4 mm) and antioxidant properties (82.3 ± 2.8%). Active fraction was loaded onto mass spectroscopy and identified the presence of four active constituents such as 1,2,9,10-tetramethoxy-6-methyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinolin-3-yl) methanol; 3',7-Dimethoxy-3-hydroxyflavone; Saponin and (19R)9acetyl19hydroxy10,14dimethyl20oxopentacyclo[11.8.0.0 < 2,10 > .0 < 4,9 > .0 < 14,19 >]henicos-17-yl-acetate. Besides, in vivo studies were conducted on Catla catla fingerlings infected with P. aeruginosa under laboratory conditions. The fingerlings were segregated into 5 groups, among which group 4 and 5 were treated with crude and purified extracts. Both the extracts were efficient in treating infected fingerlings and recorded 100% survival rate which is even better than group-3 treated with a synthetic antibiotic (77%). Hence, S. maritima leaf extract can be considered as a possible alternative medicine in aquaculture.

Impact of cypermethrin in nephrocytes of freshwater fish Catla catla.

The kidney of Catla catla, chronically exposed to sub-lethal concentrations (0.24 µg/L and 0.41 µg/L) of cypermethrin revealed a significant elevation in the activity of antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione-s-transferase (GST) and reduced glutathione (GSH) after 15 days, followed by a decline of up to 45 days. Lipid peroxidation (LPO) remained elevated throughout the exposure duration. Histology presented proliferated haematopoietic tissue, tubular and glomerular degeneration. The maximum increase in the mean degree of tissue change (DTC) was observed on the 45th day of treatment. Ultra-structure study depicted cytoplasmic vacuolation, fragmented RER, the proliferation of lysosomes, mitochondrial degeneration, and degenerative changes in the epithelial lining of renal tubules. Principal component analysis (PCA) of various biomarkers generated two components PCI (SOD, GST, GSH, LPO and DTC) and PCII (CAT). These findings suggest that long term exposure to cypermethrin can lead to various pathological alterations in the fish kidney which in turn might interfere with normal renal excretory mechanism.

NF-κB signaling induces inductive expression of the downstream molecules and IgD gene in the freshwater carp, Catla catla.

Toll-like receptors (TLRs) in innate immune system act as primary sensors in detecting the microbial components and activate their signaling cascades to induce NF-κB (nuclear factor NF-κB) towards the augmentation of immunoglobulin (Ig) synthesis. To gain insights into the efficacy of NF-κB pathway in immunoglobulin D (IgD) synthesis in the Indian Major Carp Catla catla, cloning and sequencing of TLR-signaling downstream molecules [TRAF3 (TNF receptor-associated factor 3), NEMO (nuclear factor-kappa B essential modulator), NF-κB and BAFF (B cell activating factor)] were performed by infecting the fish with pathogens. mRNA expression analysis of the downstream molecules and IgD showed significant up-regulation of these genes in kidney (P ≤ 0.001) as compared to spleen (P ≤ 0.05). To ascertain the role of NF-κB pathway in IgD synthesis, the primary cell culture of kidney and spleen in monolayer cell suspension was treated with NF-κB inhibitor (BAY 11-7082) and down-regulation of BAFF, NEMO, NF-κB, and IgD gene was observed. These results highlight the importance of NF-κB signaling pathway in augmenting the IgD gene expression in the freshwater carp, Catla catla.

Influence of IP-injected ZnO-nanoparticles in Catla catla fish: hematological and serological profile.

This study reports an effort to synthesize biocompatible zinc oxide nanoparticles using sol-gel method and its influence on hematological and serological profile of Catla catla fish. Hexagonal wurtzite structure and crystallite size of ZnO-NPs was identified by using XRD in the range of 19 to 20 nm. Moreover, the irregular and non-uniform surface of these NPs was found using SEM. The different stretched and vibrational mode (ZnO, OH, CO, and H-O-H) was identified by using FTIR analysis. UV-visible spectroscopy confirmed absorbance of the blue shift in the range 340 nm. Bioassay of ZnO-NPs on Catla catla was performed and nano ZnO was given through intraperitoneal injections at 0, 25, 50, 75, and 100 µg/g body weight doses. Analysis of fish blood samples indicated an increase in WBCs and leukocytes while the differential effect on monocytes. On the other hand in response to varying ZnO concentrations, an increase in RBCs, hemoglobin, and HCT was evident. Serum analysis revealed an increase in urea concentration while a reduction in creatinine, ALT, and AST. In an overall assessment, nano-ZnO supplementation at 25 to 100 µg/g body weight differentially affected hematological and serum biochemical profile of thaila fish. Graphical abstract.

Ontogenetic and tissue-specific expression of gonadotropin-inhibitory hormone (GnIH) and its receptors in Catla catla.

Gonadotropin-inhibitory hormone (GnIH) is an RFamide peptide, and its role in reproduction is well studied from fish to mammals, but very few reports are available about the function of GnIH during larval development. In this study, we examined the GnIH and GnIH receptors (GnIHRs) expression from embryogenesis to adult stage and tissue-specific expression in adult Catla catla using quantitative real-time (qRT) PCR. The qRT PCR analysis of GnIH mRNA during ontogenetic development showed the increasing trend from early developmental stages to the adult stage with the highest expression in 24 months fish. However, the expression of two GnIH receptors, GnIHR1 and GnIHR2 also increased from larval stages to the adults with a peak at 17 days post-hatching, while GnIHR3 showed the higher mRNA expression during embryogenesis and then decreasing gradually. Tissue distribution analysis of GnIH showed the highest mRNA expression of GnIH in the brain, followed by gonads of both the sexes. GnIHR1 and GnIHR2 were also highly expressed in the brain and gonads of both the sexes, while GnIHR3 showed the highest expression in gonads of both the sexes without any expression in the brain. These results suggest that the brain is the primary site of action for GnIH, GnIHR1 and GnIHR2, while gonads for GnIHR3.

Assessment of cypermethrin induced hepatic toxicity in Catla catla: A multiple biomarker approach.

The study was designed to evaluate chronic toxicity of pyrethroid pesticide cypermethrin through biochemical, histopathological, ultrastructural and molecular biomarkers in liver of freshwater carp Catla catla. The fish were exposed to two sub-lethal concentrations (0.21 µg/L and 0.41 µg/L) for a period of 45 days. Compared to the control, a significant (p < 0.05) increase in the activity of enzymatic antioxidants catalase (CAT), superoxide dismutase (SOD) and glutathione-S-transferase (GST), and glutathione content (GSH) was registered after initial 15 days of exposure to the toxicant, followed by decline on 30th and 45th day. Whereas, MDA level remained elevated throughout the exposure duration at both the tested concentrations. Light microscopy revealed changes like sinusoidal dilation, vacuolation, pycnosis, karyolysis, nuclear pleomorphism, lymphocyte infiltration in liver of the exposed fish, with highest mean degree of tissue change (DTC) value of 58.6 ± 3.19 on 45th day. Ultrastructurally, cytopasmic vacuolation, reduced glycogen granules, dilated RER, deformed nuclear membrane, swollen & distorted mitochondria and augmentation in lipid bodies were the prominently observed cytopathological alterations. These anomalies increased in time-concentration dependent manner, being most severe after 45 days at higher concentration. The gene expression levels of Gadd-45α and Bcl-2 depicted altered patterns. Gadd-45α exhibited significant upregulation by 45th day, while Bcl-2 demonstrated initial upregulation, with subsequent downregulation on 30th and 45th day. Principal component analysis (PCA) generated two components, PC1 (SOD, GSH, MDA and DTC) and PC2 (CAT and GST). The findings suggest that cypermethrin inflicts marked hepatototoxic effects on Catla catla even at sub-lethal concentrations.

Effect of chilled storage on quality characteristics of battered and breaded snack product from large sized Catla (Catla catla).

This study was performed to determine quality changes of battered and breaded pre-fried snack product developed from large sized (5.5-10.5 kg) Catla (Catla catla) fish stored at chilled temperature (- 2 to - 4 °C). Quality changes were assessed based on biochemical, microbiological and sensory quality evaluation at 2 days interval for a period of 18 days. Results of biochemical analysis showed increase in total volatile base-nitrogen (TVB-N) values from initial value of 4.15-13.74 mg/100 g, free fatty acid (FFA) increased from 0.95 to 2.98% in terms of oleic acid, peroxide value (PV) increased from 2.6 to 3.9 meq of O2/kg of fat and pH increased from 6.5 to 6.79 during 18 days chilled storage. Linearity (r2) of TVB-N, FFA, PV and pH values during storage were found to be 0.98, 0.99, 0.97 and 0.99 respectively. In microbiological quality evaluation total plate count (Log. Values) increased from 2.17 to 4.65, while Escherichia coli, Staphylococcus aureus, Salmonella and spore count were not detected during storage period. Results of sensory analysis reported a negative correlation with storage period concluding the shelf life of battered and breaded fish cutlet from Catla fish to be 12 days at chilled storage conditions.

Molecular characterization of gonadotropin-inhibitory hormone (GnIH) gene and effect of intramuscular injection of GnIH peptide on the reproductive axis in Catla catla.

Gonadotropin-inhibitory hormone (GnIH) plays an important role in reproduction by inhibiting the expression of gonadotropins in birds and mammals, but in fishes, it is ambiguous. In this study, we cloned 606 bp long cDNA of GnIH from Catla catla brain (cGnIH). The encoded preproGnIH peptide generated three putative peptides (cGnIH-I, -II, -III) of different size. Phylogenetic analysis of GnIH showed clustering of different peptide sequence with its orthologs in separate clades. The real-time PCR analysis showed the expression of cGnIH in brain, gonads, intestine, stomach, heart, gill and liver with the highest expression in the brain and gonads of both sexes. The basal GnIH mRNA expression was higher in spawning and spent phase of the male brain and spawning phase of the female brain. In testis, the expression was highest in spent phase, while in ovary the expression did not change significantly during reproductive phases. The in vivo experiment of cGnIH-III peptide exhibited the higher expression of HPG axis genes, lhb, fshb, cgnrh, kiss2 and kiss1r and serum hormone level of LH and FSH as soon as 3 h after the intramuscular delivery. These results suggest that the GnIH is positively involved in regulation of reproduction in HPG axis of C. catla.

Molecular approach for identification of Catla catla using mitochondrial CO1 from Pakistan.

DNA barcoding is a rapid, precise, and effective way of species identification. A short and standard target gene marker is used to create sequence profile of identified species. Specific tag or marker is used, which is derived from mitochondrial COI for identification. Effectiveness of this method axes the degree of divergence among species. Identification is necessary for their representation. In the present work, Catla catla was used to study by using Cytochrome C Oxidase 1.The genetic distances were computed, and Neighbor Joining tree was constructed based on the Kimura 2 Parameter method. GenBank and BOLD revealed definitive identity matches. Conspecific and congeneric K2P nucleotide divergence was estimated. Evolutionary tree was analyzed clearly by relating their species to phylogenetic tree, as same as species were bunched under same tree node, while species were differently clustered under distinct nodes. These findings conclude that the gene sequence may serve as a milestone for identification and phylogenetic history of related species at molecular level.

Complete mitochondrial genome sequence of Catla catla (Hamilton, 1822) from the Halda river of Bangladesh.

Catla (Catla catla) is one of the fastest-growing major carp found in South Asia as well as Bangladesh. Catla catla is the second most popular indigenous carp species in the freshwater aquaculture industry of Bangladesh due to its relatively good taste and high market price. In this study, we disclosed the complete mitochondrial genome sequence of Bangladeshi Catla fish from Halda river located in Chittagong. The circular mitogenome of Catla catla is 16,597 bp in length and nucleotide composition is AT-based (72%), contains 37 genes including 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and a D-loop (control region).