Rathore, Gaurav; Kumar, Gokhlesh; Sood, Neeraj; Kapoor, D; Lakra, W S
Serum immunoglobulins [Ig] of rohu [Labeo rohita] were purified by affinity chromatography using bovine serum albumin as capture ligand. The purified rohu Ig [r-Ig] had a molecular weight [MW] of 880 kDa as determined with gel filtration chromatography. The heavy chain of r-Ig had an MW of 77.8 kDa and that of light chain was 26.4 kDa in SDS-PAGE. Purified r-Ig was used for the production of two anti-rohu Ig monoclonal antibodies [D7 and H4] that belonged to subclass IgG2b and IgG1, respectively. Both the MAbs were specific to heavy chain of r-Ig as seen in Western blotting. Anti-rohu Ig MAb was used as a diagnostic reagent in ELISA and immunocytochemical assays to demonstrate its application for sero-surveillance and for immunological studies in rohu. A competitive ELISA was used to demonstrate the antigenic relatedness of r-Ig with whole serum Ig of other fish species. Cross reactivity of anti-rohu Ig MAb was observed with serum Ig of Catla catla and Cirrihinus mrigala. No reactivity to serum Ig of Ophiocephalus striatus and Clarias gariepinus was seen. Anti-rohu Ig MAb was found to be suitable for the detection of pathogen specific [Edwardsiella tarda] antibodies in serum of immunized rohu by an indirect ELISA. In flow cytometry using D7 MAb, the mean percentage [+/-SE] of Ig positive cells in spleen and blood of rohu were found to be 64.85% [+/-2.34] and 51.84% [+/-2.55] of gated lymphocytes, respectively. Similarly, D7 MAb also stained 52.84% [+/-1.30] and 10.5% of gated lymphocytes in kidney and thymus, respectively. The anti-rohu Ig MAbs also showed specific staining of Ig bearing cells in spleen sections by the indirect immunoperoxidase test.
Rasal, Kiran Dashrath; Chakrapani, Vemulawada; Patra, Swagat Kumar; Mohapatra, Shibani D; Nayak, Swapnarani; Jena, Sasmita; Sundaray, Jitendra Kumar; Jayasankar, Pallipuram; Barman, Hirak Kumar
The myostatin (MSTN) is a known negative growth regulator of skeletal muscle. The mutated myostatin showed a double-muscular phenotype having a positive significance for the farmed animals. Consequently, adequate information is not available in the teleosts, including farmed rohu carp, Labeo rohita. In the absence of experimental evidence, computational algorithms were utilized in predicting the impact of point mutation of rohu myostatin, especially its structural and functional relationships. The four mutations were generated at different positions (p.D76A, p.Q204P, p.C312Y, and p.D313A) of MSTN protein of rohu. The impacts of each mutant were analyzed using SIFT, I-Mutant 2.0, PANTHER, and PROVEAN, wherein two substitutions (p.D76A and p.Q204P) were predicted as deleterious. The comparative structural analysis of each mutant protein with the native was explored using 3D modeling as well as molecular-dynamic simulation techniques. The simulation showed altered dynamic behaviors concerning RMSD and RMSF, for either p.D76A or p.Q204P substitution, when compared with the native counterpart. Interestingly, incorporated two mutations imposed a significant negative impact on protein structure and stability. The present study provided the first-hand information in identifying possible amino acids, where mutations could be incorporated into MSTN gene of rohu carp including other carps for undertaking further in vivo studies.
Rasal, Kiran Dashrath; Chakrapani, Vemulawada; Patra, Swagat Kumar; Mohapatra, Shibani D.; Nayak, Swapnarani; Jena, Sasmita; Sundaray, Jitendra Kumar; Jayasankar, Pallipuram; Barman, Hirak Kumar
The myostatin (MSTN) is a known negative growth regulator of skeletal muscle. The mutated myostatin showed a double-muscular phenotype having a positive significance for the farmed animals. Consequently, adequate information is not available in the teleosts, including farmed rohu carp, Labeo rohita. In the absence of experimental evidence, computational algorithms were utilized in predicting the impact of point mutation of rohu myostatin, especially its structural and functional relationships. The four mutations were generated at different positions (p.D76A, p.Q204P, p.C312Y, and p.D313A) of MSTN protein of rohu. The impacts of each mutant were analyzed using SIFT, I-Mutant 2.0, PANTHER, and PROVEAN, wherein two substitutions (p.D76A and p.Q204P) were predicted as deleterious. The comparative structural analysis of each mutant protein with the native was explored using 3D modeling as well as molecular-dynamic simulation techniques. The simulation showed altered dynamic behaviors concerning RMSD and RMSF, for either p.D76A or p.Q204P substitution, when compared with the native counterpart. Interestingly, incorporated two mutations imposed a significant negative impact on protein structure and stability. The present study provided the first-hand information in identifying possible amino acids, where mutations could be incorporated into MSTN gene of rohu carp including other carps for undertaking further in vivo studies. PMID:27019850
Barman, Hirak Kumar; Mohanta, Ramya; Patra, Swagat Kumar; Chakrapani, Vemulawada; Panda, Rudra Prasanna; Nayak, Swapnarani; Jena, Sasmita; Jayasankar, Pallipuram; Nandanpawar, Priyanka
We previously characterized the β-actin gene promoter of Indian domesticated rohu carp (Labeo rohita) and made a reporter construct via fusion to green fluorescence protein (GFP) cDNA. In this study, the same construct was used to breed transgenic rohu fish. About 20% of the transgenic offspring showed ubiquitous expression of the reporter GFP gene. In a few of the transgenic fish, we documented massive epithelial and/or muscular expression with visible green color under normal light. The expression of GFP mRNA was higher in the muscle tissue of transgenic fish than in that of non-transgenic fish. A highly efficient nucleofection protocol was optimized to transfect proliferating spermatogonial cells of rohu using this reporter construct. The β-actin promoter also drove expressions in HEK293 (derived from human embryonic kidney cells), K562 (human leukemic cells) and SF21 (insect ovarian cells) lines. These findings imply conserved regulatory mechanisms of β-actin gene expression across eukaryotes. Furthermore, the isolated β-actin promoter with consensus regulatory elements has the potential to be used in generating transgenic carp with genes of interest and in basic biology research.
Das, Basanta Kumar; Pradhan, Jyotirmayee; Sahu, Swagatika
The study evaluated the effect of dietary doses of Euglena viridis on the immune response and disease resistance of Labeo rohita fingerlings against infection with the bacterial pathogen Aeromonas hydrophila. L. rohita fingerlings were fed with diet containing 0 (Control), 0.1 g, 0.5 g, 1.0 g Euglena powder kg(-1) dry diet for 90 days. Biochemical (serum total protein, albumin, globulin, albumin:globulin ratio), haematological (WBC, RBC, haemoglobin content) and immunological (superoxide anion production, lysozyme, serum bactericidal activity) parameters of fish were examined after 30, 60 and 90 days of feeding. Fish were challenged with A. hydrophila 90 days post-feeding and mortalities were recorded over 10 days post-infection. The results demonstrate that fish fed with Euglena showed increased levels of superoxide anion production, lysozyme, serum bactericidal activity, serum protein and albumin (P < 0.05) compared with the control group. Following challenge with A. hydrophila less survivability was observed in the control group (56.65%) than the group fed the experimental diets. The group fed 0.5 g Euglena kg(-1) dry diet showed the highest percentage survival (75%). These results indicate that Euglena stimulates the immunity and makes L. rohita more resistant to A. hydrophila infection.
Background Production of carp dominates world aquaculture. More than 1.1 million tonnes of rohu carp, Labeo rohita (Hamilton), were produced in 2010. Aeromonas hydrophila is a bacterial pathogen causing aeromoniasis in rohu, and is a major problem for carp production worldwide. There is a need to better understand the genetic mechanisms affecting resistance to this disease, and to develop tools that can be used with selective breeding to improve resistance. Here we use a 6 K SNP array to genotype 21 full-sibling families of L. rohita that were experimentally challenged intra-peritoneally with a virulent strain of A. hydrophila to scan the genome for quantitative trait loci associated with disease resistance. Results In all, 3193 SNPs were found to be informative and were used to create a linkage map and to scan for QTL affecting resistance to A. hydrophila. The linkage map consisted of 25 linkage groups, corresponding to the number of haploid chromosomes in L. rohita. Male and female linkage maps were similar in terms of order, coverage (1384 and 1393 cM, respectively) and average interval distances (1.32 and 1.35 cM, respectively). Forty-one percent of the SNPs were annotated with gene identity using BLAST (cut off E-score of 0.001). Twenty-one SNPs mapping to ten linkage groups showed significant associations with the traits hours of survival and dead or alive (P <0.05 after Bonferroni correction). Of the SNPs showing significant or suggestive associations with the traits, several were homologous to genes of known immune function or were in close linkage to such genes. Genes of interest included heat shock proteins (70, 60, 105 and “small heat shock proteins”), mucin (5b precursor and 2), lectin (receptor and CD22), tributyltin-binding protein, major histocompatibility loci (I and II), complement protein component c7-1, perforin 1, ubiquitin (ligase, factor e4b isoform 2 and conjugation enzyme e2 c), proteasome subunit, T-cell antigen receptor and
Chandra, Gyan; Saxena, Amita
An experiment was conducted in three different tanks for 45 days (T1, T2 and T3) to investigate the effect of dietary Spirulina level on growth and chemical composition of carcass in rohu, Labeo rohita. A diet having de-oiled rice bran(30%), rice polish (15%), de-oiled mustard oil cake(25%), Soya bean oil cake(10%) and de-oiled groundnut cake(20%) was provided to the control (i.e. 100% formulated feed), whereas in T2 95% formulated feed with 5% Spirulina and in T3 90% formulated feed with 10% Spirulina. Data were analysed with ANOVA (P < 0.05) and found significant. There was significant difference in the average final weight of the fish among treatments with highest in the T3 (11.28g) followed by T2 (8.32g) and T1 (7.51g). The higher SGR was recorded in T3 (2.17) followed by T2 (1.52) and T1 (1.28). There was significant difference in the average carcass composition of the fish among treatments with highest level in T3 followed by T2 and T1.
Dubey, Saurabh; Avadhani, Kiran; Mutalik, Srinivas; Sivadasan, Sangeetha Madambithara; Maiti, Biswajit; Paul, Joydeb; Girisha, Shivani Kallappa; Venugopal, Moleyur Nagarajappa; Mutoloki, Stephen; Evensen, Øystein; Karunasagar, Indrani; Munang'andu, Hetron Mweemba
Aeromonas hydrophila is a Gram-negative bacterium that causes high mortality in different fish species and at different growth stages. Although vaccination has significantly contributed to the decline of disease outbreaks in aquaculture, the use of oral vaccines has lagged behind the injectable vaccines due to lack of proven efficacy, that being from primary immunization or by use of boost protocols. In this study, the outer membrane protein W (OmpW) of A. hydrophila was cloned, purified, and encapsulated in poly d,l-lactide-co-glycolic acid (PLGA) nanoparticles (NPs) for oral vaccination of rohu (Labeo rohita Hamilton). The physical properties of PLGA NPs encapsulating the recombinant OmpW (rOmpW) was characterized as having a diameter of 370-375 nm, encapsulation efficiency of 53% and -19.3 mV zeta potential. In vitro release of rOmpW was estimated at 34% within 48 h of incubation in phosphate-buffered saline. To evaluate the efficacy of the NP-rOmpW oral vaccine, two antigen doses were orally administered in rohu with a high antigen (HiAg) dose that had twice the amount of antigens compared to the low antigen (LoAg) dose. Antibody levels obtained after vaccination showed an antigen dose dependency in which fish from the HiAg group had higher antibody levels than those from the LoAg group. The antibody levels corresponded with post challenge survival proportions (PCSPs) and relative percent survival (RPS) in which the HiAg group had a higher PCSP and RPS than the LoAg group. Likewise, the ability to inhibit A. hydrophila growth on trypticase soy agar (TSA) by sera obtained from the HiAg group was higher than that from the LoAg group. Overall, data presented here shows that OmpW orally administered using PLGA NPs is protective against A. hydrophila infection with the level of protective immunity induced by oral vaccination being antigen dose-dependent. Future studies should seek to optimize the antigen dose and duration of oral immunization in rohu in order to
Dubey, Saurabh; Avadhani, Kiran; Mutalik, Srinivas; Sivadasan, Sangeetha Madambithara; Maiti, Biswajit; Paul, Joydeb; Girisha, Shivani Kallappa; Venugopal, Moleyur Nagarajappa; Mutoloki, Stephen; Evensen, Øystein; Karunasagar, Indrani; Munang’andu, Hetron Mweemba
Aeromonas hydrophila is a Gram-negative bacterium that causes high mortality in different fish species and at different growth stages. Although vaccination has significantly contributed to the decline of disease outbreaks in aquaculture, the use of oral vaccines has lagged behind the injectable vaccines due to lack of proven efficacy, that being from primary immunization or by use of boost protocols. In this study, the outer membrane protein W (OmpW) of A. hydrophila was cloned, purified, and encapsulated in poly d,l-lactide-co-glycolic acid (PLGA) nanoparticles (NPs) for oral vaccination of rohu (Labeo rohita Hamilton). The physical properties of PLGA NPs encapsulating the recombinant OmpW (rOmpW) was characterized as having a diameter of 370–375 nm, encapsulation efficiency of 53% and −19.3 mV zeta potential. In vitro release of rOmpW was estimated at 34% within 48 h of incubation in phosphate-buffered saline. To evaluate the efficacy of the NP-rOmpW oral vaccine, two antigen doses were orally administered in rohu with a high antigen (HiAg) dose that had twice the amount of antigens compared to the low antigen (LoAg) dose. Antibody levels obtained after vaccination showed an antigen dose dependency in which fish from the HiAg group had higher antibody levels than those from the LoAg group. The antibody levels corresponded with post challenge survival proportions (PCSPs) and relative percent survival (RPS) in which the HiAg group had a higher PCSP and RPS than the LoAg group. Likewise, the ability to inhibit A. hydrophila growth on trypticase soy agar (TSA) by sera obtained from the HiAg group was higher than that from the LoAg group. Overall, data presented here shows that OmpW orally administered using PLGA NPs is protective against A. hydrophila infection with the level of protective immunity induced by oral vaccination being antigen dose-dependent. Future studies should seek to optimize the antigen dose and duration of oral immunization in rohu in order to
Dash, P; Patel, S; Dixit, A; Garg, L C; Sahoo, P K
Cytokines are important components of both adaptive and innate immunity, and are required to initiate and regulate immune responses following infection. The ontogeny and tissue specific distribution of four pro-inflammatory cytokines, interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), IL-8 and IL-1β in rohu (Labeo rohita), and their responses by leucocytes from anterior-kidney/head-kidney (HKLs), spleen (SPLs) and peripheral blood (PBLs) following stimulation with concanavalin A (ConA), ConA with phorbol 12-myristate 13-acetate (ConA/PMA) and formalin-killed Aeromonas hydrophila cells (FAH) were studied. In ontogeny study, mRNA levels of IL-6 and IL-1β were evident in unfertilized egg stages of L. rohita whereas IL-8 and TNF-α transcripts were found from 1 to 3 h post-fertilization (hpf) onwards till day 15 post-fertilization, respectively. Basal level of all four cytokines was observed in all twelve tissues (eye, brain, heart, gill, anterior kidney, posterior kidney, spleen, liver, skin, muscle, hindgut and foregut) of L. rohita juveniles. Expression levels of IL-6 and IL-8 were found to be the highest in liver and heart tissues, respectively, while TNF-α transcripts were high in anterior kidney and liver tissues. Transcripts of IL-1β showed high expression in muscle, heart and spleen. Upon in vitro stimulation of leucocytes, there was variable up-regulation of all the four cytokines following different treatments throughout the experimental time period. Induction of cytokines was more pronounced in PBLs stimulated with FAH compared to other stimuli. However, an up-regulated IL-8 expression was evident in all the leucocytes following stimulation with FAH thus indicating IL-8 could be used as an indicator or indirect marker to monitor vaccine status or health status of L. rohita during bacterial infection.
Sahu, Dinesh Kumar; Panda, Soumya Prasad; Meher, Prem Kumar; Das, Paramananda; Routray, Padmanav; Sundaray, Jitendra Kumar; Jayasankar, Pallipuram; Nandi, Samiran
Rohu is a leading candidate species for freshwater aquaculture in South-East Asia. Unlike common carp the monsoon breeding habit of rohu restricts its seed production beyond season indicating strong genetic control over spawning. Genetic information is limited in this regard. The problem is exacerbated by the lack of genomic-resources. We identified 182 reproduction-related genes previously by Sanger-sequencing which were less to address the issue of seasonal spawning behaviour of this important carp. Therefore, the present work was taken up to generate transcriptome profile by mRNAseq. 16GB, 72bp paired end (PE) data was generated from the pooled-RNA of twelve-tissues from pre-spawning rohu using IlluminaGA-II-platform. There were 64.97 million high-quality reads producing 62,283 contigs and 88,612 numbers of transcripts using velvet and oases programs, respectively. Gene ontology annotation identified 940 reproduction-related genes consisting of 184 mainly associated with reproduction, 223 related to hormone-activity and receptor-binding, 178 receptor-activity and 355 embryonic-development related-proteins. The important reproduction-relevant pathways found in KEGG analysis were GnRH-signaling, oocyte-meiosis, steroid-biosynthesis, steroid-hormone biosynthesis, progesterone-mediated oocyte-maturation, retinol-metabolism, neuroactive-ligand-receptor interaction, neurotrophin-signaling and photo-transduction. Twenty nine simple sequence repeat containing sequences were also found out of which 12 repeat loci were polymorphic with mean expected-&-observed heterozygosity of 0.471 and 0.983 respectively. Quantitative RT-PCR analyses of 13-known and 6-unknown transcripts revealed differences in expression level between preparatory and post-spawning phase. These transcriptomic sequences have significantly increased the genetic-&-genomic resources for reproduction-research in Labeo rohita. PMID:26148098
Sahu, Dinesh Kumar; Panda, Soumya Prasad; Meher, Prem Kumar; Das, Paramananda; Routray, Padmanav; Sundaray, Jitendra Kumar; Jayasankar, Pallipuram; Nandi, Samiran
Rohu is a leading candidate species for freshwater aquaculture in South-East Asia. Unlike common carp the monsoon breeding habit of rohu restricts its seed production beyond season indicating strong genetic control over spawning. Genetic information is limited in this regard. The problem is exacerbated by the lack of genomic-resources. We identified 182 reproduction-related genes previously by Sanger-sequencing which were less to address the issue of seasonal spawning behaviour of this important carp. Therefore, the present work was taken up to generate transcriptome profile by mRNAseq. 16 GB, 72 bp paired end (PE) data was generated from the pooled-RNA of twelve-tissues from pre-spawning rohu using IlluminaGA-II-platform. There were 64.97 million high-quality reads producing 62,283 contigs and 88,612 numbers of transcripts using velvet and oases programs, respectively. Gene ontology annotation identified 940 reproduction-related genes consisting of 184 mainly associated with reproduction, 223 related to hormone-activity and receptor-binding, 178 receptor-activity and 355 embryonic-development related-proteins. The important reproduction-relevant pathways found in KEGG analysis were GnRH-signaling, oocyte-meiosis, steroid-biosynthesis, steroid-hormone biosynthesis, progesterone-mediated oocyte-maturation, retinol-metabolism, neuroactive-ligand-receptor interaction, neurotrophin-signaling and photo-transduction. Twenty nine simple sequence repeat containing sequences were also found out of which 12 repeat loci were polymorphic with mean expected-&-observed heterozygosity of 0.471 and 0.983 respectively. Quantitative RT-PCR analyses of 13-known and 6-unknown transcripts revealed differences in expression level between preparatory and post-spawning phase. These transcriptomic sequences have significantly increased the genetic-&-genomic resources for reproduction-research in Labeo rohita.
Ramachandran, S; Bairagi, A; Ray, A K
Eight isonitrogenous (35% crude protein approximately) and isocaloric (4.0 kcalg(-1) approximately) diets were formulated incorporating raw and fermented grass pea (Lathyrus sativus) seed meal at 10%, 20%, 30% and 40% levels by weight into a fish meal based diet and fed to rohu, Labeo rohita, fingerlings for 80 days and fish performance was studied. A particular bacterial strain (Bacillus sp.) isolated from the intestine of adult common carp (Cyprinus carpio) reared in the wild having significant amylolytic, cellulolytic, lipolytic and proteolytic activities were used for fermentation of seed meal for 15 days at 37 degrees C. Fermentation of grass pea seed meal was effective in significantly reducing the crude fibre content and anti-nutritional factors, such as tannins, phytic acid and the neurotoxin, beta-ODAP and enhancing the available free amino acids and fatty acids. In terms of growth response, feed conversion ratio and protein efficiency ratio, 30% fermented grass pea seed meal incorporated diet resulted in significantly (P < 0.05) better performance of rohu fingerlings. In general, growth and feed utilization efficiencies of fish fed diets containing fermented seed meal were superior to those fed diets containing raw seed meal. The apparent protein digestibility (APD) values decreased with increasing levels of raw seed meal in the diets. The APD for raw seed meal was lower at all levels of inclusion in comparison to those for the fermented seed meals. The highest deposition of carcass protein was recorded in fish fed the diet containing 40% fermented seed meal. The results indicated that fermented grass pea seed meal can be incorporated in carp diets up to 30% level compared to 10% level of raw seed meal.
Giri, Sib Sankar; Jun, Jin Woo; Sukumaran, Venkatachalam; Park, Se Chang
To explore the feasibility of Musa acuminata (banana) peels as a feed additive, effects of banana peel flour (BPF) on the growth and immune functions of Labeo rohita were evaluated. Diets containing five different concentrations of BPF (0% [basal diet], 1% [B1], 3% [B3], 5% [B5], and 7% [B7]) were fed to the fish (average weight: 15.3 g) for 60 days. The final weight gain and specific growth rate were higher (P < 0.05) in the B5 group. The most significant improvements in immune parameters such as lysozyme, alternative complement pathway, leukocyte phagocytic, superoxide dismutase, and catalase activities were observed in the B5 group. However, the B5 group exhibited the lowest malondialdehyde activity. IgM and glutathione peroxidise activities were significantly elevated in the treatment groups, except in B1, after only 30 days of feeding. Of the examined cytokine-related genes, IL-1β, TNF-α, and HSP70 were upregulated in the head kidney and hepatopancreas, and expressions were generally higher in the B3 and B5 groups. Moreover, B5 group challenged with Aeromonas hydrophila 60 days after feeding exhibited the highest survival rate (70%; P < 0.05). These results suggest that dietary BPF at 5% could promote growth performance and strengthen immunity in L. rohita.
Giri, Sib Sankar; Jun, Jin Woo; Sukumaran, Venkatachalam; Park, Se Chang
To explore the feasibility of Musa acuminata (banana) peels as a feed additive, effects of banana peel flour (BPF) on the growth and immune functions of Labeo rohita were evaluated. Diets containing five different concentrations of BPF (0% [basal diet], 1% [B1], 3% [B3], 5% [B5], and 7% [B7]) were fed to the fish (average weight: 15.3 g) for 60 days. The final weight gain and specific growth rate were higher (P < 0.05) in the B5 group. The most significant improvements in immune parameters such as lysozyme, alternative complement pathway, leukocyte phagocytic, superoxide dismutase, and catalase activities were observed in the B5 group. However, the B5 group exhibited the lowest malondialdehyde activity. IgM and glutathione peroxidise activities were significantly elevated in the treatment groups, except in B1, after only 30 days of feeding. Of the examined cytokine-related genes, IL-1β, TNF-α, and HSP70 were upregulated in the head kidney and hepatopancreas, and expressions were generally higher in the B3 and B5 groups. Moreover, B5 group challenged with Aeromonas hydrophila 60 days after feeding exhibited the highest survival rate (70%; P < 0.05). These results suggest that dietary BPF at 5% could promote growth performance and strengthen immunity in L. rohita. PMID:27294156
Rauta, Pradipta Ranjan; Nayak, Bismita
Advanced vaccine research approaches needs to explore on biodegradable nanoparticles (NPs) based vaccine carrier that can serve as antigen delivery systems as well as immuno-stimulatory action to induce both innate and adaptive immune response in fish. Immunogenicity of PLA and PLGA NPs encapsulating outer membrane protein (Omp) antigen of Aeromonas hydrophila were evaluated through intra-peritoneal injection in fish, Labeo rohita. Antigen loaded PLA-Omp (223.5 ± 13.19 nm) and PLGA-Omp (166.4 ± 21.23 nm) NPs were prepared using double emulsion method by efficiently encapsulating the antigen reaching the encapsulation efficiency 44 ± 4.58% and 59.33 ± 5.13% respectively. Our formulated PLA Omp and PLGA-Omp NPs were in nanometer range (<500 nm) and could be successfully endocyted in the body. Despite low antigen loading in PLA-Omp, it showed considerably slower antigen release in vitro than PLGA-Omp NPs. Other physical properties like zetapotential values and poly dispersity index (PDI) confirmed the stability as well as monodisperse nature of the formulated nanoparticles. The spherical and isolated nature of PLA-Omp and PLGA-Omp NPs were revealed by SEM analysis. Upon immunization of all antigenic formulations (PLA-Omp NP, PLGA-Omp NP, FIA-Omp, PLA NP, PLGA NP, PBS as control), significant higher bacterial agglutination titre and haemolytic activity were observed in case of PLA-Omp and PLGA-Omp immunized groups than rest groups at both 21 days and 42 days. The specific antibody response was significantly increased and persisted up to 42 days of post immunization by PLA-Omp, PLGA-Omp, FIA-Omp. PLA-Omp NPs showed better immune response (higher bacterial agglutination titre, haemolytic activity, specific antibody titre, higher percent survival upon A. hydrophila challenge) than PLGA-Omp in L. rohita confirming its better efficacy. Comparable antibody response of PLA-Omp and PLGA-Omp with FIA-Omp treated groups suggested that PLA and PLGA could be replacement for
Sethuramalingam, T A; Haniffa, M A
Six sets of feeding experiments were carried out using formulated diets containing prawn head waste (PW), chicken intestine waste (CW), banana flower (BF), cauliflower waste (CAU) Dolicos lab lab (DLL) and groundnut leaf (GNL) in four levels of inclusion (15, 30, 45 and 60%) to assess the pattern of distribution and activities of digestive enzymes like cellulase, amylase, maltase, invertase, protease and lipase in the digestive tracts of Labeo rohita fingerlings. A control group of fish was fed with diets containing antibiotics to destroy the digestive tract microflora which may induce digestive functions. In general, the activity of digestive enzymes depended on the amount and type of the ingredients present in the diets ingested by the fish. Test animals showed both endogenous and bacterial cellulase activities which suggests the necessity for including cellulose (plant protein source) as dietary ingredient. Occurrence of higher amount of cellulase in the foregut and amylase in the fore and midgut influenced by DNL and GNL diets revealed the possibility of including less than 40% of the respective ingredients in the diet of rohu. Maltase and invertase were highly influenced by GNL, DLL and BF diets than PW and CW diets. More than 40% inclusion of PW and CW was found to increase protease and lipase secretion in the midgut and hindgut regions. The higher secretion of lipase in the midgut suggested the physiological versatility for lipid digestion in rohu fingerlings.
Zutshi, Bela; Prasad, S G Raghu; Nagaraja, R
Blood is an indicator of physiological condition of an animal. Therefore, a field study was conducted to investigate the hematological parameters of wild population of rohu, Labeo rohita (Ham). The following aspects were evaluated in blood: hemoglobin content, red blood cell (RBC) and white blood cell (WBC) count, packed cell volume (PCV), and mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) values, and in plasma: cholesterol, protein, and glucose levels. For this purpose, rohu fish of varying sizes and weights were sampled from Hebbal (receiving a storm water drain) and Chowkalli lake (received domestic sewage and industrial effluents from various sources and was more polluted than Hebbal lake). It revealed noticeable differences in hemoglobin content, RBC and WBC count, and PCV and MCHC values. Severe anemia can be marked by a significant decrease in RBC count (p < 0.5), hemoglobin content, and PCV and MCHC values, whereas an increase in leukocyte count and MCV values were observed in fish from Chowkalli lake. Fish from lake B had fewer RBC and low concentration of serum protein and cholesterol. Serum concentration of glucose showed initial higher levels and then low concentration (900-1,500 g) in fish from lake B when compared to lake A. The variation in values of different parameters can be attributed to exposure of fish to various types of pollutants present mainly in the Chowkalli lake which receives heavy metals, synthetic detergents, petroleum products, and other acid and alkali substances from the nearby local industries. Other observations of these fish include dark body color and aggressive nature of fish.
Mishra, Prabhakar; R S, Suresh Kumar; Jerobin, Jayakumar; Thomas, John; Mukherjee, Amitava; Chandrasekaran, Natarajan
Presence of several biochemical constituents in neem makes it an efficient antimicrobial agent for pathogenic diseases. The current investigation was aimed to assess the therapeutic potential of neem nanoemulsion as a control measure for Pseudomonas aeruginosa infection in freshwater fish Labeo rohita. The median lethal concentration (LC50) for the neem oil and neem nanoemulsion was 73.9 and 160.3 mg/L, respectively. The biomarker enzymes of treated fish tissues showed a significant difference in the level of glutathione reductase, catalase, and lipid peroxidation in neem oil-treated samples than in neem nanoemulsion-treated samples at P<0.05. The results were corroborative with histopathology and ultrastructural analysis. The bacterial infection of P. aeruginosa treated using neem nanoemulsion was more effective in both in vitro and in vivo methods. Present findings suggest that neem-based nanoemulsion has negligible toxicity to Rohu fishes. This makes neem-based nanoemulsion as an efficient therapeutic agent against P. aeruginosa infection, leading to its possible usage in the aquaculture industry.
Ismail, Muhammad; Khan, Qaiser Mahmood; Ali, Rahat; Ali, Tayyaba; Mobeen, Ameena
Chlorpyrifos is a widely used insecticide of organophosphate group, which causes severe toxicological effects in non target aquatic organisms especially in fish. In the present study the genotoxic effects of sublethal concentrations of chlorpyrifos were observed in the erythrocytes and gill cells of Labeo rohita (commonly known as rohu) using the Alkaline Single-Cell Gel Electrophoresis (Comet) assay. Effects of chlorpyrifos on the behavior of the fish were also investigated. The 96 h LC50 value of chlorpyrifos, estimated by Trimmed Spearman-Karber (TSK) in static bioassay, was found to be 442.8 µg/L. On the basis of LC50 value, the fish were exposed to three sublethal concentrations of chlorpyrifos (SL-I ∼221.4 µg/L, SL- II ∼110.7 µg/L and SL-III ∼73.8 µg/L) for 96 h. Blood and gill samples were collected at every 24 h and were subjected to the Comet assay. The observed DNA damage was concentration dependent and time dependent and those levels of DNA damage in between the tested concentrations and times were significantly different (p < 0.01). It was also found that the gill cells are more sensitive to chlorpyrifos, though; it revealed more DNA damage as compared to the erythrocytes of fish. Fish exposed to different concentrations of chlorpyrifos showed different neurotoxic behavioral responses. It was concluded that chlorpyrifos is a genotoxic and neurotoxic insecticide causing DNA damage and neurotoxic effects in Labeo rohita.
Giri, Sib Sankar; Sen, Shib Sankar; Jun, Jin Woo; Sukumaran, Venkatachalam; Park, Se Chang
The present study investigated the protective effects of leucine against lipopolysaccharide (LPS)-induced inflammatory responses in Labeo rohita (rohu) in vivo and in vitro. Primary hepatocytes, isolated from the hepatopancreas, were exposed to different concentrations of LPS for 24 h to induce an inflammatory response, and the protective effects of leucine against LPS-induced inflammation were studied. Finally, we investigated the efficiency of dietary leucine supplementation in attenuating an immune challenge induced by LPS in vivo. Exposure of cells to 10-25 μg mL(-1) of LPS for 24 h resulted in a significant production of nitric oxide and release of lactate dehydrogenase to the medium, whereas cell viability and protein content were reduced (p < 0.05). LPS exposure (10 μg mL(-1)) increased mRNA levels of the pro-inflammatory cytokines TNF-α, IL-1β and IL-8 in vitro (p < 0.05). However, pretreatment with leucine prevented the LPS-induced upregulation of TNF-α, IL-1β and IL-8 mRNAs by downregulating TLR4, MyD88, NF-κBp65, and MAPKp38 mRNA expression. Interestingly, mRNA expression of the anti-inflammatory cytokine, IL-10, which was increased by LPS treatment, was further enhanced (p < 0.05) by leucine pretreatment. The enhanced expression of IL-10 might inhibit the production of other pro-inflammatory cytokines. It was found that leucine pretreatment attenuated the excessive activation of LPS-induced TLR4-MyD88 signaling as manifested by lower level of TLR4, MyD88, MAPKp38, NF-κBp65 and increased level of IκB-α protein in leucine pre-treatment group. In vivo experiments demonstrated that leucine pre-supplementation could protect fish against LPS-induced inflammation through an attenuation of TLR4-MyD88 signaling pathway. Taken together, we propose that leucine pre-supplementation decreases LPS-induced immune damage in rohu by enhancing the expression of IL-10 and by regulating the TLR4-MyD88 signaling pathways.
Mitra, Gopa; Mukhopadhyay, P K; Ayyappan, S
The effect of supplementation of ascorbic acid through enriched zooplankton [10%, 20% and 30% ascorbyl palmitate (AP) inclusion in diet of zooplankton] on different digestive enzyme activities during ontogeny of Labeo rohita larvae was studied from 4 day to 15 day post hatch. Ascorbic acid (AA) content in different groups of unenriched (8.6+/-0.71) and enriched zooplankton were, 750+/-29.3, 1409.1+/-45.5, 2009.21+/-199.2 mug/g respectively on dry matter basis with differences (P<0.05) between the treatments. A difference (P<0.05) was found in tissue AA level in different dietary groups. Low amylase, protease, lipase and alkaline phosphatase activities were present in rohu larvae from the mouth opening stage which showed increasing trend with the age of larvae and increasing dietary AA content. A clear dose-dependent modulation of digestive enzyme activities in response to 10%, 20% and 30% AP enriched zooplankton feeding was evidenced from positive correlations between dietary AA content with magnitude of elevation of enzyme activity in different groups. There were 57, 55, 29.2 and 2 fold increases in amylase activity; 7.35, 7.02, 4.43 and 2.73 fold increases in protease activity; 45.636, 41.50, 19.83 and 13.69 fold increases in lipase activity and 6, 5, 3, and 2 fold increases in alkaline phosphatase activity observed in the 15th day post hatch larvae fed 20%, 30%, 10%AP enriched and normal zooplankton respectively, than 4-day post hatch larvae of the respective groups. Enzyme activities were also positively correlated with specific growth rates of wet weight of rohu larvae at the 15th day post hatch. Increased AA might have played an important role in advancing morphological transformation of the digestive tract, protecting gastric mucosa and accelerating growth by the process of tissue formation, which necessitated the requirement of more nutrient thereby, increasing digestive enzyme activity. The regulatory role of AA in the modulation of different digestive
Population structure and genetic diversity of Indian Major Carp, Labeo rohita (Hamilton, 1822) from three phylo-geographically isolated riverine ecosystems of India as revealed by mtDNA cytochrome b region sequences.
Behera, Bijay Kumar; Baisvar, Vishwamitra Singh; Kunal, Swaraj Priyaranjan; Meena, Dharmendra Kumar; Panda, Debarata; Pakrashi, Sudip; Paria, Prasenjit; Das, Pronob; Bhakta, Dibakar; Debnath, Dipesh; Roy, Suvra; Suresh, V R; Jena, J K
The population structure and genetic diversity of Rohu (Labeo rohita Hamilton, 1822) was studied by analysis of the partial sequences of mitochondrial DNA cytochrome b region. We examined 133 samples collected from six locations in three geographically isolated rivers of India. Analysis of 11 haplotypes showed low haplotype diversity (0.00150), nucleotide diversity (π) (0.02884) and low heterogeneity value (0.00374). Analysis of molecular variance (AMOVA) revealed the genetic diversity of L. rohita within population is very high than between the populations. The Fst scores (-0.07479 to 0.07022) were the indication of low genetic structure of L. rohita populations of three rivers of India. Conspicuously, Farakka-Bharuch population pair Fst score of 0.0000, although the sampling sites are from different rivers. The phylogenetic reconstruction of unique haplotypes revealed sharing of a single central haplotype (Hap_1) by all the six populations with a point mutations ranging from 1-25 nucleotides.
Karan, Sweta; Dash, Pujarini; Kaushik, Himani; Sahoo, Pramoda K.; Garg, Lalit C.
Interleukin-10, an important regulator of both the innate and adaptive immune systems, is a multifunctional major cytokine. Though it is one of the major cytokines, IL-10 from the Indian major carp, Labeo rohita, has not yet been characterized. In the present study, we report large scale production and purification of biologically active recombinant IL-10 of L. rohita (rLrIL-10) using a heterologous expression system and its biophysical and functional characterization. High yield (~70 mg/L) of soluble rLrIL-10 was obtained at shake flask level. The rLrIL-10 was found to exist as a dimer. Far-UV CD spectroscopy showed presence of predominantly alpha helices. The tertiary structure of the purified rLrIL-10 was verified by fluorescence spectroscopy. Two-dimensional gel analysis revealed the presence of six isoforms of the rLrIL-10. The rLrIL-10 was biologically active and its administration significantly reduced serum proinflammatory cytokines, namely, interleukin 1β, TNFα, and IL-8, and augmented the NKEF transcript levels in spleen of L. rohita. Anti-inflammatory role of the rLrIL-10 was further established by inhibition of phagocytosis using NBT reduction assay in vitro. The data indicate that the dimeric alpha helical structure and function of IL-10 of L. rohita as a key regulator of anti-inflammatory response have remained conserved during evolution. PMID:27689097
Tiwari, Sudhanshu; Tiwari, Richa; Singh, Ajay
Laboratory evaluations were made to assess the toxicological and biochemical effect of cypermethrin on fingerlings of common edible freshwater culture carp (Labeo rohita). There was a significant negative (P < 0.05) correlation observed between effective doses of cypermethrin and exposure periods; that is, LC50 values decreased from 0.323 μg/L (6 h) to > 0.278 μg/L (12 h), > 0.240 μg/L (18 h) and >0.205 μg/L (24 h). Exposure to sublethal doses of cypermethrin for 24 h and 96 h exposure period caused significant (P < 0.05) time- and dose-dependent alterations in total protein, total free amino acids, nucleic acids, glycogen, pyruvate, and lactate level and in the activity of enzyme protease, alanine aminotransferase, aspartate aminotransferase, acid phosphatases, alkaline phosphatases, acetylcholinesterase, and cytochrome oxidase in liver and muscle tissues of fish. Thus, cypermethrin has potent piscicidal activity against fingerlings of fish Labeo rohita and adversely affects their behavioural patterns, shifting aerobic pathway of fish respiration towards anaerobic pathway and also inhibiting energy production by suppressing ATP synthesis. PMID:22666103
Maharana, Jitendra; Swain, Banikalyan; Sahoo, Bikash R; Dikhit, Manas R; Basu, Madhubanti; Mahapatra, Abhijit S; Jayasankar, Pallipuram; Samanta, Mrinal
In lower eukaryotes-like fish, innate immunity contributed by various pattern recognition receptor (PRR) plays an essential role in protection against diseases. Nucleotide-binding and oligomerization domain (NOD)-2 is a cytoplasmic PRR that recognizes MDP (muramyl dipeptide) of the Gram positive and Gram negative bacteria as ligand and activates signalling to induce innate immunity. Hypothesizing a similar NOD2 signalling pathway of higher eukaryotes, the peripheral blood leucocytes (PBLs) of rohu (Labeo rohita) was stimulated with MDP. The data of quantitative real-time PCR (qRT-PCR) revealed MDP-mediated inductive expression of NOD2 and its down-stream molecule RICK/RIP2 (receptor-interacting serine-threonine protein kinase-2). This observation suggested the existence of MDP-binding sites in rohu NOD2 (rNOD2). To investigate it, 3D model of ligand-binding leucine-rich repeat (LRR) region of rNOD2 (rNOD2-LRR) was constructed following ab initio and threading approaches in I-TASSER web server. Structural refinement of the model was performed by energy minimization, and MD (molecular dynamics) simulation was performed in GROMACS (Groningen Machine for Chemical Simulations). The refined model of rNOD2-LRR was validated through SAVES, ProSA, ProQ, WHAT IF and MolProbity servers, and molecular docking with MDP was carried out in GOLD 4.1. The result of docking identified LRR3-7 comprising Lys820, Phe821, Asn822, Arg847, Gly849, Trp877, Trp901 and Trp931 as MDP-binding critical amino acids in rNOD2. This is the first study in fish to provide an insight into the 3D structure of NOD2-LRR region and its important motifs that are expected to be engaged in MDP binding and innate immunity.
Akhtar, M S; Pal, A K; Sahu, N P; Ciji, A; Meena, D K; Das, P
Two experiments were conducted to elucidate the possible effects of dietary L-tryptophan (TRP) in Labeo rohita based on growth performance and physio-biochemical responses. In the experiment I, a 60-day feeding trial was carried out to elucidate the effects of dietary TRP enrichment on growth performance and physio-biochemical responses. In the experiment II, the TRP pre-fed L. rohita, from experiment I, was exposed to temperature and salinity stress to evaluate stress-mitigating efficacy of TRP. In L. rohita, dietary supplementation of TRP showed significant effect on weight gain percentage and feed conversion ratio but not on blood glucose. A significant increase in RNA content and RNA/DNA ratio upon TRP supplementation was observed and was positively correlated with growth performance. The results of experiment II indicated that weight gain percentage, serum T3 and T4 levels were significantly reduced in groups that were exposed to temperature and salinity stress and fed diets without TRP supplementation. However, dietary supplementation of TRP significantly augmented weight gain percentage in stress-exposed groups. Tryptophan supplementation helped in bringing back T3 and T4 levels comparable with control. A significant increase in superoxide dismutase, catalase, Adenosine triphosphatase, blood glucose and serum cortisol was observed in temperature- and salinity-exposed groups fed without TRP-supplemented diets. However, TRP supplementation was found to be effective in restoring the above parameters. The results of these experiments suggest that dietary TRP supplementation augments growth, lowers energy demand and helps in mitigating thermal and salinity stress in L. rohita.
Alexander, C; Sahu, N P; Pal, A K; Akhtar, M S; Saravanan, S; Xavier, B; Munilkumar, S
A 60-day experiment was conducted to delineate the effect of three dietary levels of gelatinized carbohydrate (GC) on growth, nutrient-utilization and body composition of Labeo rohita fingerlings (avg. wt 6.5 ± 0.3 g) reared at two water temperatures (ambient-AT (26 ± 0.8 °C) and 32 °C). Two hundred and sixteen fingerlings were randomly distributed into six treatments in triplicates. Three semi-purified isonitrogenous diets were prepared with graded levels of GC viz. D(1) : 40%, D(2) : 50% and D(3) : 58%. Growth rate, feed efficiency and protein efficiency ratio were significantly (p < 0.05) higher in 50% GC and 32 °C reared groups than their AT counterparts. Hepato Somatic Index was higher in AT reared groups compared to 32 °C reared counterparts. Apparent digestibility co-efficient of carbohydrate was significantly (p < 0.05) higher at 32 °C reared groups but decreased with increasing carbohydrate (GC) levels. Fish reared at 32 °C showed significantly (p < 0.05) higher amylase, protease and hexokinase activities while glucose-6-phosphate dehydrogenase and glucose-6-phosphatse were significantly (p < 0.05) higher at ambient temperatures. The results obtained in present study indicated that L. rohita could utilize higher level (50%) of dietary carbohydrate at 32 °C.
Samanta, Mrinal; Swain, Banikalyan; Basu, Madhubanti; Mahapatra, Girishbala; Sahoo, Bikash R; Paichha, Mahismita; Lenka, Saswati S; Jayasankar, Pallipuram
Toll-like receptors (TLRs) are a class of innate immune receptors that sense pathogens or their molecular signatures and activate signaling cascades to induce a quick and non-specific immune response in the host. Among various types of TLRs, TLR22 is exclusively present in teleosts and amphibians and is expected to play the distinctive role in innate immunity. This report describes molecular cloning, three-dimensional (3D) modeling, and expression analysis of TLR22 in rohu (Labeo rohita), the most commercially important freshwater fish species in the Indian subcontinent. The open reading frame (ORF) of rohu TLR22 (LrTLR22) comprised of 2,838 nucleotides (nt), encoding 946 amino acid (aa) residues with the molecular mass of ∼ 107.6 kDa. The secondary structure of deduced LrTLR22 exhibited the presence of signal peptide (1-22 aa), 18 leucine-rich repeat (LRR) regions (79-736 aa), and TIR domain (792-935 aa). The 3D model of LrTLR22-LRR regions together elucidated the horse-shoe-shaped structure having parallel β-strands at the concave surface and few α-helices at the convex surface. The TIR domain structure revealed alternate presence of five α-helices and β-sheets. Phylogenetically, LrTLR22 was closely related to common carp and exhibited significant similarity (92.2 %) and identity (86.1 %) in their amino acids. In rohu, TLR22 was constitutively expressed in all embryonic developmental stages, and tissue-specific analysis illustrated its expression in all examined tissues, highest was in liver and lowest in brain. In vivo modulation of TLR22 gene expression was analyzed by quantitative real-time PCR (qRT-PCR) assay following stimulation with lipopolysaccharide (LPS), synthetic double stranded RNA (polyinosinic-polycytidylic acid), and bacterial (Aeromonas hydrophila) RNA. Among these ligands, bacterial RNA most significantly (p < 0.05) induced TLR22 gene expression in most of the tested tissues. In A. hydrophila infection, induction of TLR22 gene expression
Ullah, Farhad; Bari, Fathul; Raziq, Sumayya; Riaz, Mehreen; Khan, Khalid; Nishan, Umar; Sthanadar, Iram Alam; Shaheen, Baseerat; Shaheen, Mussarrat; Ahmad, Habib
We examined and compared heavy metals bioaccumulation in Cyprinus carpio and Labeo rohita netted from Sardaryab, a tributary of River Kabul. By using atomic absorption spectrometry we assessed different organs including livers, gills, and muscles. Metals studied were chromium, iron, zinc, lead, and copper. Livers of both species showed higher concentrations of metals while muscles showed the least amount. Chromium and iron were the highly concentrated metals in the gills and livers of both species. A quantity of 0.154 ± 0.011, 0.199 ± 0.0079, and 0.024 ± 0.008 μg/g of chromium was found in the gills, livers, and muscles of Cyprinus carpio, respectively. Similarly, the gills, liver, and muscles of Labeo rohita contained 0.133 ± 0.008, 0.165 ± 0.01, and 0.019 ± 0.006 μg/g of Cr, respectively. Iron was highest in carp in the range of 0.086 ± 0.01 in gills and 0.067 ± 0.011 μg/g in muscles, comparatively. All the studied metals were found within the US recommended daily dietary allowances (RDA) limits; hence no immediate risk in their consumption for human was found. The data showed that Cyprinus carpio being omnivorous and bottom feeder stored higher concentrations of metals as compared to Labeo rohita.
The Bacillus licheniformis strain isolated from the intestine of Labeo rohita by an enrichment technique showed capability of utilizing dimethoate as the sole source of carbon. The bacterium rapidly utilized dimethoate beyond 0.6 mg/mL and showed prolific growth in a mineral salts medium containing 0.45 mg/mL dimethoate. The isolated B licheniformis exhibited high level of tolerance of dimethoate (3.5 mg/mL) in nutrient broth, while its cured mutant did not tolerate dimethoate beyond 0.45 mg/mL and it was unable to utilize dimethoate. The wild B licheniformis strain transferred dimethoate degradation property to E coli C600 (Nar, F−) strain. The transconjugant harbored a plasmid of the same molecular size (approximately 54 kb) as that of the donor plasmid; the cured strain was plasmid less. Thus a single plasmid of approximately 54 kb was involved in dimethoate degradation. Genes encoding resistance to antibiotic and heavy metal were also located on the plasmid. PMID:16192686
Jindal, Rajinder; Verma, Sakshi
Cadmium chloride (CdCl2) induced genotoxicity and cytotoxicity has been assessed in the peripheral blood erythrocytes of freshwater fish Labeo rohita exposed to 0.37 and 0.62mg/L of CdCl2 in water for 100 days. The blood samples of the fish were collected at different intervals (days 1, 3, 5, 10, 15, 30, 60 and 100) of exposure period to analyze DNA damage using comet assay and the occurrence of micronuclei and other cellular anomalies. The results of comet assay showed a significant increase in the mean percentage of tail DNA at both the concentrations. Exposure to CdCl2 also induced micronuclei in addition to many nuclear abnormalities such as nuclear bud, binucleates, lobed, notched and vacuolated nuclei. Cytoplasmic abnormalities like echinocytes, acanthocytes, notched, microcytes and cells with vacuolated cytoplasm were also observed. The metal exposed groups showed significant variation in the frequency of cellular abnormalities as well as the extent of DNA damage in comparison to controls. These frequencies increased significantly (p<0.05) in concentration dependent manner, peaking on 10th day while a decreasing trend was observed after 15 days of the exposure period.
Upadhyay, Jayti; Jauhari, R K; Pemola Devi, N
On examining 144 specimens of Labeo rohita (Ham.) for parasites occurrence; as many as 34 fish were found positive to harbour ciliophorans (Trichodina sp. and Ichthyophthirius multifilis Fouquet, 1876), monogeneans (Haplocleidus vachi Tripathi, 1959 and Dactylogyrus glossogobii Jain, 1960), trematode (Allocreadium mahaseri Pandey, 1939), nematode (Camallanus (Zeylanema) anabantis Pearse, 1933) and acanthocephalan (Sachalinorhynchus sp). Highest prevalence was shown by platyhelminthes (18.75%) followed by ciliophorans (10.41%), nematode (4.16%) and acanthocephalan (2.77%) in succession. Maximum mean intensity and abundance has been shown by C. (Zeylanema) anabantis Pearse, 1933 while the monogenean H. vachi was on the 2nd rank. On the other hand, the least intensity and abundance was shown by Ichthyophthirius multifilis. The prevalence was recorded more in monsoon (52.77%) followed by post-monsoon (22.22%), summer (19.44%) and winter (8.33%) in succession. Further, the unit-wise intensity of parasites was recorded more in males than the females.
Dube, Praveen N; Hosetti, Basaling B
Present study concerns the effect of sodium cyanide on the Indian major carp, Labeo rohita. Fishes were exposed to lethal (0.32 mg/L) and sublethal (0.064 mg/L) concentrations of sodium cyanide. The effect of intoxication was studied on Na(+)K(+)ATPase, Mg(+2)ATPase and Ca(2+)ATPase in various physiological tissues (gill, liver, and muscle) at the end of 1, 2, 3 and 4 days of lethal and 5, 10 and 15 days of sublethal exposure periods. Sodium cyanide induced significant inhibitory effects on the ATPase activity of the fish. Inhibition of the ATPase blocked the active transport system of the gill epithelial as well as chloride cells, and thus altered the osmo-regulatory mechanism of the fish. The value of the measured responses as an indicator of stress caused by water contamination discussed. The results confirm that ATPase levels significantly decreased in treated fish, indicating that ATPases could be used as sensitive and useful biomarkers for cyanide pollution.
Khare, Ankur; Dange, Swati
The evaluation of metal's toxicity in freshwater is one of the imperative areas of research and there is an emergent concern on the development of techniques for detecting toxic effects in aquatic animals. Oxidative stress biomarkers are very useful in assessing the health of aquatic life and more in depth studies are necessary to establish an exact cause effect relationship. Therefore, to study the effectiveness of this approach, a laboratory study was conducted in the fish Labeo rohita as a function of hexavalent chromium and the toxicity indices using a battery of oxidative stress biomarkers such as catalase (CAT), superoxide dismutase (SOD), and glutathione reductase (GR) in the liver, muscle, gills, and brain have been studied along with biometric parameters, behavioral changes, and Cr bioaccumulation. A significant increased HSI was observed in contrast to CF which reduced significantly. SOD, CAT, and GR activity increased significantly in all the tissues of treated fishes. The bioaccumulation of Cr was highest in liver followed by gills, muscle, and brain. This study highlights the significance of using a set of integrated biomarker and advocate to include these parameters in National Water Quality Monitoring Program in areas potentially polluted with metals to assess the health of the ecosystem. PMID:25302308
Mondal, S; Mondal, A; Mandal, N; Mondal, B; Mukhopadhyay, S S; Dey, A; Singh, S
The chemically treated Labeo rohita scale is used for synthesizing hydroxyapatite (HAp) biomaterials. Thermogravimetric and differential thermal analyses of fish scale materials reveal the different phase changes with temperature and find out the suitable calcination temperatures. The composition and structures of wet ball-milled calcined HAp powders are characterized by Fourier transform infrared spectroscopy, X-ray diffraction, field emission scanning electron microscopy, transmission electron microscopy, energy dispersive X-ray analysis (EDX). The EDX as well as chemical analysis of fish scale-derived apatite materials confirms that the Ca/P ratio is 1.71. The compressive stress, hardness and porosity have been evaluated on sintered HAp biomaterials. The cell attachment on HAp surfaces, cytotoxicity evaluation and MTT assay, which are carried out in RAW macrophage-like cell line media demonstrate good biocompatibility. The histological analysis also supports the bioaffinity of processed HAp biomaterials in Wistar rat model for investigating the contact reaction and stability at the artificial or natural prosthesis interface.
Yadav, Manoj K; Pradhan, Pravata K; Sood, Neeraj; Chaudhary, Dharmendra K; Verma, Dev K; Debnath, Chandan; Sahoo, Lopamudra; Chauhan, U K; Punia, Peyush; Jena, Joy K
The fish pathogenic oomycete Aphanomyces invadans is the causative agent of epizootic ulcerative syndrome (EUS), a fish disease of international significance and reportable to the World Organisation for Animal Health. In spite of the current and potential impact of A. invadans infection on fisheries and aquaculture sectors of the world, very little is known about the host-A. invadans interactions. In the present study, following experimental infection with A. invadans in one of the Indian major carps, Labeo rohita, sequential changes in various innate immune parameters were monitored. The results indicated that at early stages of infection, no significant changes in any of the studied innate immune parameters were observed. However, at the advanced stages of infection from 6 to 12 days post infection (dpi), the respiratory burst and alternate complement activity were significantly higher whereas lysozyme, antiproteases and α-2 macroglobulin values were significantly lower than the control group and also from the infected group at earlier stages of infection. Since, the possibility of vaccination of fish against A. invadans appears remote due to difficulties in eliciting a specific antibody response, the information generated in the present study could be useful for developing strategies for improving resistance to A. invadans infection by stimulating the innate immunity through immunomodulation.
Rajkumar, K. S.; Kanipandian, N.; Thirumurugan, R.
The increasing use of nano based-products induces the potential hazards from their manufacture, transportation, waste disposal and management processes. In this report, we emphasized the acute toxicity of silver nanoparticles (AgNPs) using freshwater fish Labeo rohita as an aquatic animal model. The AgNPs were synthesized using chemical reduction method and the formation of AgNPs was monitored by UV-Visible spectroscopy analysis. The functional groups, crystaline nature and morphological characterizations were carried out by fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and high resolution transmission electron microscopy (HRTEM) analysis. UV-Vis range was observed at 420 nm and XRD pattern showed that the particles are crystalline nature. HRTEM analysis revealed that the morphology of particles was spherical and size ranges between 50 and 100 nm. This investigation was extended to determine the potential acute toxicity, L. rohita was treated orally with the lethal concentration (LC50) of AgNPs. The antioxidative responses were studied in the three major tissues such as gill, liver and muscle of L. rohita. The results of this investigation showed that increasing the concentration of AgNPs led to bioaccumulation of AgNPs in the major tissues. The haematological parameters showed significant alterations in the treated fish. The histological changes caused by chemically synthesized AgNPs demonstrated the damages in the tissues, primary lamella and blood vessels of L. rohita. The histological study also displayed the formation of vacuolation in liver and muscle when compared with untreated tissues (control) of L. rohita.
Sahu, Swagatika; Das, Basanta Kumar; Pradhan, Jyotirmayee; Mohapatra, B C; Mishra, B K; Sarangi, Niranjan
The study evaluated the efficacy of dietary doses of Mangifera indica (mango) kernel on the immune response and disease resistance of Labeo rohita fingerlings against the bacterial pathogen Aeromonas hydrophila infections in. L. rohita fingerlings fed diet containing 0 (Control), 1g, 5 g, 10 g mango kernel kg(-1) dry diet for 60 days. Biochemical (serum total protein, albumin, globulin, albumin:globulin ratio, blood glucose), haematological (WBC, RBC, haemoglobin content) and immunological (superoxide anion production, lysozyme, serum bactericidal activity) parameters of fish were examined at 20, 40 and 60 days of feeding. Fish were challenged with A. hydrophila 60 days post feeding and mortalities were recorded over 10 days post-infection. The results demonstrate that fish fed with mango kernel showed enhanced superoxide anion production, lysozyme, serum bactericidal, serum protein, albumin (P<0.05) compared with the control group. The mortality (%) was recorded up to 10 th day post-challenge. Less survivability was observed in control group (50%) up to day 10 after infection. The survivability was higher in experimental diets. The group fed 5 g kernel kg(-1) dry diet showed highest percentage survival (98%). These results indicate that mango kernel stimulates the immunity and makes L. rohita more resistant to A. hydrophila infection.
Palaniappan, PL. RM.; Renju, V. B.
Heavy metal pollution is a major environmental problem in the modern world due to increasing human activities. Zinc is an essential element involved in a wide variety of cellular processes. However, it becomes toxic when elevated concentrations are introduced into the environment. The goal of the present study is to investigate the effect of zinc exposure on the biochemical contents of the muscle tissues of freshwater species Labeo rohita using Fourier transform infrared (FT-IR) spectroscopy. Since the muscle constitutes the greatest mass of the fish that is consumed, the present study has paid particular attention to muscle component. The result reveals that the zinc exposure causes significant changes in the biochemical contents of the L. rohita muscle tissues. In addition, it causes an alteration in the protein secondary structures by decreasing the α-helix and increasing the β-sheet contents of muscle tissues. Further, it has been observed that the administration of chelating agent D-penicillamine improves the protein and lipid contents in the muscle tissues compared to zinc exposed tissues. This result shows that D-penicillamine is the effective chelator of zinc in reducing the body burden of L. rohita fingerlings.
Palaniappan, PL. RM.; Vijayasundaram, V.
Arsenic compounds are ubiquitous and widespread in the environment as a result of natural or anthropogenic occurrence. Fish are the major source of protein for human consumption. They are also a source of contamination, because of the amounts of heavy elements they can contain, some of which are highly toxic. Fish bones are high in calcium, which is an essential mineral for normal body function. It consists of water, organic material, and mineral matter. Chelating agents have been used clinically as antidotes for acute and chronic metal intoxications. In the present study, an attempt is made to investigate the bio-accumulation of arsenic and its effect on the biochemical and mineral contents of Labeo rohita bones using, Fourier transform infrared (FT-IR) spectroscopy. The results of the present study indicate that arsenic exposure induces significant reduction on the biochemical and mineral contents of the L. rohita bones. Further, the DMSA treatment significantly improves these levels. This shows that DMSA is an effective chelator for arsenic toxicity. Quantitative curve-fitting analyses of amide I band have proved useful in studying the nature and the extent of protein conformational changes. A decrease in α-helical and random coil structures and an increase in β-sheet structures have been observed due to arsenic exposure. In conclusion, the present study shows that the FT-IR spectroscopy coupled with second derivative and curve-fitting analysis gives useful information about the biochemical and mineral contents of the L. rohita bones.
Mir, Javaid Iqbal; Sarkar, Uttam Kumar; Gusain, Om Prakash; Dwivedi, Arvind Kumar; Joukrushna, Jena
Labeo rohita is a member of the Indian major carp species and originally an inhabitant of the Ganga river network in India. It is among the top ten aquaculture species of the world. Since there is a lack of information on the growth pattern of the wild populations of this species, this study aimed at evaluating the pattern of age and growth, to support the development of effective management plans. A total of 1082 samples of L. rohita were obtained from May 2009 to July 2012 in six drainages of the Ganga basin. Scales of the fish were used to determine the age and growth by analyzing the annual rings growth. Out of six populations, a maximum 8+ age classes were recorded from two rivers (Betwa and Sharda). The back-calculated lengths at 8+ age class ranged from 86.22 cm to 91.66 cm. However, for the rest of rivers up to 7+ age classes were recorded. Among growth parameters, specific rate of length increase (C(l)) and specific rate of weight increase (Cw) showed decreasing trend, and three distinct life stages of L. rohita were recorded based on growth constant data (C(lt)). Analysis of variance (ANOVA) of the back-calculated length data set of six populations indicated a significant difference (p < 0.05). All three populations showed significant variation in length attainment during 1+ and 3+ age groups, while two populations showed significant variation in length attainment during the 2+ and 7+ age classes. Additionally, analysis of age frequency at different length intervals indicated that with increase in age class, number of fish samples was reduced. Since the pattern of life history traits of L. rohita have not been attempted in the recent past; therefore, this study will guide fisheries biologists about the current stock structure of this fish across different spatial scale of the Ganga basin.
Xiao, Wu-Han; Xie, Xing-Ren; Gong, Xiao-Ning
The early stages and capsulogenetic stages of sporogenesis of Myxobolus kwangsiensis were studied in detail by electron microscopy. M. kwangsiensis is an intracellular myxosporean infecting the epithelial cells of kidney tubules of young dace Labeo rohita. The M. kwangsiensis sporogensis presents the typical features of myxosporea. The initial stages are multinucleate primary cells, with one primary cell typically enclosing one secondary cell. The secondary cells are formed directly by cytokinesis inside the primary cells. The primary cell participates in the formation of the secondary cell nucleus. The nuclear division of primary cell is asynchronous by amitosis. The parasitic organisms may degenerate inside the epithelial cells at any stage. The modes of sporoblast formation, the typical features of myxosporea and the cause of its degradation are discussed.
Ashok, A; Rao, D S; Raghuramulu, N
This investigation was directed towards finding the need of vitamin D for fish. The freshwater column feeder fish Labeo rohita (Rora) was used for the study. Early fry stage fish were divided into four experimental groups of 350 each: two groups were kept in natural light, while the other two were maintained under total darkness. One each of the light and dark-grown groups was supplied dietary vitamin D3 [1,650 i.u/kg diet], whereas the other groups were fed a vitamin D-deficient diet for six months. The known vitamin D-related functions and growth parameters were studied in these four experimental groups of fish. The results showed that fish reared on vitamin D-deficient diet and in dark did not have even traces of liver vitamin D, indicating a state of vitamin D deficiency in these fish. No significant differences were observed in percent bone to body weight or dry matter of the vitamin D-deficient/supplemented groups of fish grown in light/dark. Further, it was also observed that there were no significant changes in bone and carcass ash or calcium and phosphorus content in response to vitamin D3 supplementation as compared to the groups which did not receive vitamin D3 (grown both in light and dark). Also, there was little change in several other parameters like carcass protein and lipid, mortality rates, hepatosomatic index, and feed efficiency between the vitamin D-deficient/supplemented groups of fish. Thus, these findings suggest that vitamin D may not be an essential nutrient for Rora (Labeo rohita) as a representative of freshwater fish.
Saravanan, M.; Suganya, R.; Ramesh, M.; Poopal, R. K.; Gopalan, N.; Ponpandian, N.
The wide use of iron oxide nanoparticles (Fe2O3 NPs) in various applications has raised great concerns worldwide. In this work, we measured the potential harmful effects of Fe2O3 NP (<50 nm) at concentrations of 1 and 25 mg/L on haematological, biochemical, and ionoregulatory responses in an Indian major carp, Labeo rohita for a short-term period of 96 h. The results revealed significant ( P < 0.05) decreases in haemoglobin, haematocrit, mean cellular volume, mean cellular haemoglobin, protein, sodium (Na+), potassium (K+), chloride (Cl-) and gill Na+/K+-ATPase levels in both the concentrations. White blood cell, mean cellular haemoglobin concentration and glucose levels were significantly ( P < 0.05) increased in response to both concentrations during the study period. However, no significant changes in red blood cell count and gill Na+/K+-ATPase (25 mg/L) activity were noticed compared to those of the respective control groups. Based on this study, it was found that the Fe2O3 NPs do have prominent effects on freshwater fish L. rohita. Our data suggest that the alterations of these parameters can be used as nonspecific biomarkers to monitor the environmental risks arising from nanoparticles in aquatic ecosystem and also regulate the use, production and release of nanoparticles.
Sharma, Niti; Rather, Mohd Ashraf; Ajima, Malachy N O; Gireesh-Babu, P; Kumar, Kundan; Sharma, Rupam
The increasing application of silver nanoparticles (Ag-NPs) both in industries and in agricultural fields has led to its accumulation in the aquatic ecosystem through water run-off. In the present study, the effects of Ag-NPs in the liver of Labeo rohita, were investigated at genomic and cellular level for seven days at the concentrations of 100, 200, 400 and 800 μg l(-1) by using 18 and 29 nm sizes of Ag-NPs. The Ag-NPs sizes of 18 and 29 nm were synthesized by a chemical method using atomic force microscopy with the zeta potential of -55 mV and-31.4 mV respectively. They were found to be spherical with smooth surfaces. Assessment of genotoxic effects of the particles in the fish using single-cell gel electrophoresis showed DNA damage on exposure to concentrations of 400 and 800 μg l(-1). Histopathological examination of the liver revealed vacuolar degeneration, hepatocytes have undergone total degeneration and high accumulation of Ag-NPs that depicted both time and dose-dependent relationships. Furthermore, the expression study of stress-related genes showed down-regulation, due to the production of free radicals and reactive oxygen species. Ag-NPs can cause both DNA damage and affect the cellular responses of L. rohita.
Dube, Praveen N; Hosetti, B B
The effects of exposure to one-third and one-fifth sublethal concentrations (0.106 and 0.064 mg/L) of sodium cyanide on protein metabolism on freshwater carp, Labeo rohita, was studied. Three functionally different tissues, namely, the liver, muscle, and gills, were studied after 5, 10, and 15 days. Exposures produced marked changes in protein metabolic profile in all tissues studied. These changes were more pronounced in the one-third sublethal concentration, suggesting a cumulative action of toxicant. This investigation revealed that the total, structural, and soluble proteins and urea content in all the three tissues were decreased, whereas free amino acids, ammonia, and enzyme activity (i.e., protease, alanine aminotransferase, and aspartate aminotransferase) exhibited elevated levels at both sublethal concentrations. Variation in protein metabolism in the fish, induced by sodium cyanide, demonstrated its toxic effects on cellular metabolism, thereby leading to impaired protein synthetic machinery. The results of the present study indicate that a mechanism of impaired energy transformation has direct action on the fish, L. rohita, and its impact is clearly evident from the change in the nutritional content of the fish.
Giri, Sib Sankar; Sen, Shib Sankar; Chi, Cheng; Kim, Hyoun Joong; Yun, Saekil; Park, Se Chang; Sukumaran, V.
The present study aimed to investigate the effects of Chlorophytum borivilianum polysaccharide (CBP), as a dietary supplement administered at varying concentrations with feed (basal diet), on various cytokine-related responses in Labeo rohita fingerlings. Immune parameters and immune-related gene expressions were measured at 3rd, 4th, and 5th week after feeding. The results revealed that dietary administration of CBP at 0.2% and 0.4% for 4 weeks significantly upregulated serum lysozyme and phagocytic activity. Complement C3 and respiratory burst activity (RBA) were significantly higher after 4 weeks of CBP feeding. The immune related genes IL-8, IL-1β, TNF-α, and iNOS were downregulated (P < 0.05) in groups with 0.2% and 0.4% CBP supplemented diets at week 4. Expression of anti-inflammatory cytokines (IL-10 and TGF-β) was also downregulated (P < 0.5) after 4 weeks of feeding with 0.2% to 0.8% CBP. However, five weeks of CBP administration had no significant effect on immune gene expression, except TNF-α and IL-8. Fish fed with 0.4% CBP for 4 weeks showed maximum resistance against Aeromonas hydrophila (73.3% survival) compared to control. From these results, we recommend that CBP administration at 0.4% for 4 weeks could effectively improve immune response and disease resistance in L. rohita. PMID:26649325
Bindu, M S; Sobha, V
Impact of three different types of seaweed diets on growth, feed utilization and nutrient digestibility of L. rohita was studied for 120 days. The seaweed diet fed fishes, especially Ulva based diet showed comparatively higher growth and weight increment. Good food conversion ratio, food assimilation efficiency, protein efficiency ratio and better nutrient digestibility were recorded for seaweed diet fed fishes. The results suggests the suitability of utilizing seaweeds, Ulva fasciata, Spyridia insignis and Sargassum wightii as partial substitute for fishmeal in formulated diets of L. rohita.
Ghosh, Debasree; Mandal, Dipak Kumar
Bioaccumulation of mercury and histomorphological changes in the olfactory epithelium of Labeo rohita were investigated after exposing the fish to two sublethal concentrations of HgCl₂ (66 and 132 μg/L) for 15 and 30 days. Mercury deposition increased in the tissue significantly (p < 0.05) with dose- and duration-dependent manner. Severe damage to the olfactory epithelium was evident. When fish exposed to 66 μg/L for 15 days, the histology of olfactory epithelium exhibited that mucous cell proliferation was upregulated and cell size was significantly increased from the control. Similar trends were found in 30 days exposure in both treated groups. Histology showed that mercury induced degeneration of columnar sensory cells, supporting cells and ciliated non-sensory cells and induced basal cell proliferation. Basal cell hyperplasia led to form intraepithelial proliferative lesion, thickening of epithelium, basal lamina disruption and cyst formation. Scanning electron microscopy revealed that mercury exposure at 66 μg/L caused clumping and loss of cilia, erosion in microridges on the supporting cells and proliferation of mucous cell opening. Complete degeneration of ciliated cells and cyst formation was observed in the fish when exposed to 132 μg/L HgCl₂. This result suggests that prolonged exposure to mercury might cause irreversible damage to the olfactory epithelium and impair the olfactory function of fish.
Sumathi, C; Dillibabu, V; Madhuri, Dash-Koney; Priya, D Mohana; Nagalakshmi, C; Sekaran, G
Abstract: This study stresses the key role which can be played by Tannery Fleshing (TF) hydrolyzing probiotic Pontibacter spp. in aqua feed formulation and identifies the probiotic strains in the fish gut capable of enhancing the overall growth and immune responses. Probiotics included are Pontibacter species (Pb) and Bacillus megaterium (BM) wherein Lactobacillus (LB) served as control. Experimental diets includes tannery fleshing (TF1), TF+LB strain (TF2), TF+BM strain (TF3), TF+Pb strain (TF4), Fishmeal+BM(TF5), Fishmeal+Pb and Control fish meal based diet (TF6). Compared with control, total weight gain (TWG), Specific Growth Rate (SGR), Feed Conversion Ratio (FCR) and Protein Efficiency Ratio (PER) in fish fed with diets supplemented with probiotics were significantly increased (p < 0.05). NBT, lysozyme activity, total protein and globulin content were highest in TF4 diet. After challenge with Aeromonas hydrophila, TF4 recorded highest survival and TF1 lowest survival in comparison with the control. Growth and related parameters reveals the effective utilization potential of tannery fleshing probiotic as a feed source. Comparative studies with standard fish meal diets reveals that the fish fed with Pontibacter spp. and Bacillus megaterium included feeds enhanced both assimilating capacity and immunological responses in Labeo rohita.
Vaseem, Huma; Banerjee, T K
A field study was conducted to examine different physicochemical properties of water and various haematological and biochemical parametres of the fish Labeo rohita collected from the Ganga River (National river of India) at Varanasi district, India. The water was found to be greatly contaminated with a number of dissolved metals (Fe, Cr, Zn, Cu, Mn, Ni and Pb) whose concentrations were above the safe limits suggested by Bureau of Indian Standard (BIS 1991) for drinking water (Fe, 1,353.33 %; Cr, 456 %; Mn, 553.33 %; Ni, 4,490 % and Pb, 1,410 %). The metal accumulation in the fish blood was very high (Fe, 2,408 %; Cr, 956.57 %; Zn, 464.90 %; Cu, 310.57 %; Mn, 1,115.48 %) in comparison to the control fish maintained under strict quality control. Lower values of the various haematological parameters (total erythrocytes count, haemoglobin, haematocrit, mean corpuscular volume and O2-carrying capacity) in the river fish in comparison to the control indicate toxic manifestation exerted by the contaminated river water on the fish. The higher level of total leucocytes count further illustrates stressed condition of the river fish. The toxic impact of the Ganga water is also expressed in the fish by the presence of higher levels of cholesterol, glucose, elevated activities of the enzymes aspartate amino transferase and alanine amino transferase, and lowered protein concentration.
Alexander, C; Sahu, N P; Pal, A K; Akhtar, M S
A feeding trial of 70-days was carried out to study the haemato-immunological and stress responses of Labeo rohita fingerlings reared at two water temperatures [ambient (Amb) - 27 °C and 32 °C] fed with graded levels of gelatinized corn carbohydrate (GC). Two hundred and sixteen fingerlings were randomly distributed into six treatment groups in triplicate. Three semi-purified diets were prepared containing 30% crude protein with graded levels of GC 40%, 50% and 58%. The six treatment groups were T(1) (40% GC × Amb), T(2) (40% GC × 32 °C), T(3) (50% GC × Amb), T(4) (50% GC× 32 °C), T(5) (58% GC × Amb) and T(6) (58% GC × 32 °C). The blood glucose level was significantly (p < 0.05) lowered in groups fed with 58% GC level. Neither dietary GC levels nor temperature had a significant (p > 0.05) effect on serum cortisol and superoxide dismutase activity. Lysozyme activity was significantly higher (p < 0.05) in T(1) during pre- and post-challenge period while temperature alone had a significant (p < 0.05) effect on post-challenge Nitroblue Tetrazolium and found higher at 32 °C. A significant effect of GC levels and rearing temperature was recorded on WBC in the pre- and post-challenge period. Highest pre-challenge WBC was observed in T(4) group and in the post-challenge period T(1) group recorded maximum. Water temperature had significant effect on pre-challenge haemoglobin content, highest being at 32 °C (T(2) ). A significant (p < 0.05) effect of rearing temperature and dietary GC level on total serum protein and albumin was also observed. Highest total serum protein and albumin was recorded in T(1) and globulin in T(2) . Percentage survival after challenging with Aeromonas hydrophila was highest in T(1) followed by T(3) group and lowest in T(6) . The results obtained in the present study suggest that L. rohita fingerlings may utilize higher levels of dietary GC at higher temperature (32 °C) but may affect its immunity status.
Palaniappan, PL. RM.; Nishanth, T.; Renju, V. B.
In the present work, an attempt has been made to assess the bioconcentration and distribution of zinc on the selected organs of Labeo rohita and to study the effect of zinc exposure on the biochemical constitutions of gill tissues of L. rohita by using FT-IR Spectroscopy. The concentration pattern in the organs reveals that the liver is the prime site of metal binding and muscle accumulates least metal concentration. The accumulation profile is in the order: liver > gill > kidney > brain > bone > muscle. It has also been observed that the administration of chelating agent D-Penicillamine (DPA) reduces the zinc concentration in all tissues more effectively than the administration of the chelating agent Ethylene Diamine Tetra Acetic acid. The FT-IR spectra reveal that zinc exposure causes significant changes in the biochemical constitutions of the gill tissues. It causes an alteration in the protein secondary structures by decreasing the α-helix and increasing the β-sheet contents. Further, it has been observed that the administration of chelating agent DPA improves the protein and lipid contents in the gill tissues compared to zinc exposed tissues. This result shows that DPA is the effective chelator of zinc in reducing the body burden of L. rohita fingerlings. In conclusion, the findings of the current study suggest that zinc exposure causes significant changes in both lipids and proteins of the gill tissues, and changes the protein profile in favour of β-sheet structure.
Giri, Sib Sankar; Sen, Shib Sankar; Sukumaran, V
The effects of dietary Pseudomonas aeruginosa VSG-2 supplementation on innate immunity and protection against Aeromonas hydrophila infection were evaluated in Labeo rohita. Fish were fed for 60 days with control diet or 3 experimental diets containing P. aeruginosa VSG-2 at 10(5), 10(7), and 10(9) cfu g(-l), respectively. Various innate immune parameters were examined at 30 and 60 days post-feeding. Fish were challenged with A. hydrophila 60 days post-feeding and mortalities were recorded over 10 days post-infection. Dietary supplementation of P. aeruginosa VSG-2 significantly increased serum lysozyme and alternative complement pathway (ACP) activities, phagocytosis, and respiratory burst activity in head kidney macrophages of L. rohita throughout the experimental period. Superoxide dismutase (SOD) activity significantly increased after 60 days in the groups fed diets containing 10(7) and 10(9) cfu g(-1) P aeruginosa. Serum IgM levels were significantly higher in the treatment groups than in the control group after 30 days of feeding; however, the opposite result was observed at 60 days. Moreover, fish fed diets containing 10(7) and 10(9) cfu g(-1)P. aeruginosa had significantly higher post-challenge survival rates against A. hydrophila infection. Further, P. aeruginosa VSG-2 was found to be safe for mammals. These results indicate that dietary P. aeruginosa VSG-2 supplementation at 10(7) cfu g(-1) can effectively improve innate immunity and disease resistance in L. rohita.
Sukumaran, Venkatachalam; Park, Se Chang; Giri, Sib Sankar
This study evaluated the effects of ginger (Zingiber officinale) as a feeding supplement on the growth, skin mucus immune parameters, and cytokine-related gene expression of Labeo rohita, and its susceptibility to Aeromonas hydrophila infection. Diets containing six different concentrations of dried ginger (0% [basal diet], 0.2% [G2], 0.4% [G4], 0.6% [G6], 0.8% [G8], and 1.0% [G10] were fed to fish (average weight: 12.3 g) for 60 days. Growth parameters were examined at 30 and 60 days post-feeding. Skin mucosal immune responses and gene expression were examined 60 days post-feeding. Results showed that growth parameters such as final weight gain (93.47 ± 1.73 g) and specific growth rate (3.41 ± 0.14) were significantly higher in G8 than in the control. Among the skin mucosal immune parameters examined, lysozyme (46.5 ± 3.8 U mg(-1)), immunoglobulin level (8.9 ± 0.4 unit-mg mL(-1)), protein level (44.3 ± 2.2 mg mL(-1)) were significantly higher in G8. However, alkaline phosphatase activity (171.6 ± 10.2 IU L(-1)) was high (P < 0.05) in the G10 group. Skin mucus of G8 exhibited significantly higher inhibition zones when tested against pathogenic bacterial strains. For cytokine-related genes, anti-oxidant genes (zinc/copper superoxide dismutase [SOD1], glutathione peroxidase [GPx], anti-inflammatory cytokines (interleukin-10 [IL-10], transforming growth factor-beta [TGF-β]), signalling molecules nuclear factor erythroid 2-related factor 2 [Nrf2], and Inhibitor protein κBα [IκB-α]) were all up-regulated in the head kidney, intestine, and hepatopancreas of fish that were fed experimental diets. In addition, expression abundance was significantly higher in most tissues in G2 and/or G10, than in the control. Conversely, expression of genes encoding pro-inflammatory cytokines (IL-1β, tumour necrosis factor-alpha [TNF-α]), signalling molecules Kelch-like-ECH-associated protein 1 (Keap1), and nuclear factor kappa B p65 (NF-κBp65) were down
Sharma, Arun; Deo, Ashutosh D; Riteshkumar, S Tandel; Chanu, Thongam Ibemcha; Das, Arabinda
The present study evaluated the efficacy of dietary doses of Withania somnifera (Ashwagandha) root powder on immunological parameters and disease resistance against the bacterial pathogen Aeromonas hydrophila infections in Indian major carp, Labeo rohita fingerlings. Fishes were fed with dry diet containing 0 gkg(-1) (control), 1 gkg(-1) (T(1)), 2 gkg(-1) (T(2)) and 3 gkg(-1) (T(3)) W. somnifera root powder for 42 days. Immunological (NBT level, Phagocytic activity, total immunoglobulin and lysozyme activity) parameters of fishes were examined at 0 days, 14 days, 28 days and 42 days of feeding. Fishes were challenged with A. hydrophila 42 days post feeding and mortalities (%) were recorded over 14 days post-infection. The results demonstrate that fishes fed with W. somnifera root showed enhanced NBT level, Phagocytic activity, total Immunoglobulin level and lysozyme activity (p<0.05) compared with the control group. The survivability was higher in experimental diets than the control group. Dietary W. somnifera at the level of 2 gkg(-1) showed significantly (P<0.05) higher protection (RPS 42.85+/-0.65%) against A. hydrophila infection than control. The results suggest that the W. somnifera root powder have a stimulatory effect on immunological parameters and increases disease resistance in L. rohita fingerlings against A. hydrophila infection.
Ramesh, Dharmaraj; Vinothkanna, Annadurai; Rai, Amit Kumar; Vignesh, Venkada Subramanian
Bacillus species isolated from the gut of healthy Labeo rohita (Hamilton) were screened for antibacterial activity against selected fish pathogens. Among the isolates, KADR5 and KADR6 showed antibacterial activity, tolerated low pH and high bile concentrations and were susceptibility to various antibiotics. Based on morphological and biochemical tests and 16S rRNA gene analysis the probiotic strains KADR5 and KADR6 were identified as Bacillus licheniformis and Bacillus pumilus, respectively. The immune stimulatory effect of subcellular components of probiotic Bacillus licheniformis KADR5 and Bacillus pumilus KADR6 in L. rohita against Aeromonas hydrophila infection was studied. Fish were immunized intraperitoneally in case of subcellular components [cell wall proteins (CWPs), extracellular proteins (ECPs), whole cell proteins (WCPs)] and orally in case of live cells (10(8) CFU/g of feed). After 14th day of administration, fishes from each group were challenged intraperitoneally with 0.1 ml of A. hydrophila cell suspension in PBS (10(5) cells ml(-1)). Groups immunized with subcellular components and live cells had significantly lower mortalities of 20-40% and 23-33%, respectively in comparison to control (80% mortality). The non specific immune factors in the cellular components and viable cells of the probiotics increased the expression of lysozyme and respiratory burst. Use of WCPs and CWPs resulted in better protection against A. hydrophila in L. rohita. Our results clearly reflect the potential of cellular components of the probiotics Bacillus species for the protection of fish against A. hydrophila infection by enhancing the immune response.
Giri, Sib Sankar; Sen, Shib Sankar; Chi, Cheng; Kim, Hyoun Joong; Yun, Saekil; Park, Se Chang; Sukumaran, V
The present investigation evaluated the effects of dietary leucine (Leu) on growth performance, head kidney antioxidant status, and gene expression in Labeo rohita juveniles. Fish were fed with six isonitrogenous diets containing graded levels of Leu at 0.75 (control), 1.7, 3.2, 4.6, 6.3, and 7.6 g Leu kg(-1) of feed for 8 weeks. Compared with the control group, appropriate Leu supplementation significantly enhanced the percent weight gain (PWG), feed intake (FI), and protein efficiency ratio (PER) (P<0.05) but decreased the plasma ammonia content (PAC) (P<0.05). Similarly, optimal Leu supplementation stimulated head kidney glutathione (GSH) content, and superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities as compared to the control group; however, a reverse trend was observed in malondialdehyde (MDA) content. Further, relative gene-expression levels of lysozyme, complement C3, β-microglobulin, immunoglobulin-M, SOD, GPx, nuclear factor erythroid 2-related factor 2 (Nrf2), natural killer-cell enhancing factor β (NKEF-β), and toll-like receptor-22 (TLR22) in the head kidney were enhanced (P<0.05) at leucine levels of 4.6 g kg(-1) of feed. Conversely, the mRNA levels of tumour necrosis factor-α (TNF-α), Kelch-like-ECH-associated protein 1 (Keap1), and interleukin 1β (IL-1β) in head kidney were down-regulated by Leu supplementation. Collectively, our results revealed that appropriate Leu supplementation improved fish growth and antioxidant capacity, and regulated the mRNA levels of related signalling molecules in L. rohita juveniles. Based on the quadratic regression analysis of PWG, PER, and PAC, the optimum dietary leucine requirements of L. rohita juveniles were estimated to be 4.7, 4.5, and 4.8 g kg(-1) of feed.
Kumar, Shivendra; Sahu, N P; Gal, Denes
The present study was conducted to investigate the strategy to mitigate the immunosuppressive and oxidative stress effect of gelatinized starch in fingerling of Labeo rohita. Fingerlings were either maintained at ambient water temperature (26 °C) or exposed to 32 °C for one week and then subjected to 26 °C for four weeks. Both groups were fed with isoproteinous (30% crude protein) diets containing gelatinized (G) or non-gelatinized (NG) starch. After 5 weeks of feeding trial, fingerlings were challenged by Aeromonas hydrophila and survival rate was recorded for the next 7 days. Serum cortisol and glucose content was significantly (p < 0.05) higher in G starch fed group and decreased with the increase in temperature from 26 to 32 °C, which was consistent for next four week after decrease in temperature from 32 to 26 °C. Lower respiratory burst activity and serum total protein and globulin content in G starch fed group at 26 °C significantly (p < 0.05) increased after elevation of temperature from 26 to 32 °C and levelled off to NG starch fed group. Liver superoxide dismutase (SOD) and catalase (CAT) activity of G starch fed group was significantly higher in group reared at 32 °C compared to 26 °C. After challenge, fish fed G starch showed lower survival rate than that of fish fed NG starch. Subsequently, exposure of elevated temperature (32 °C) for one week significantly increased the survival rate of G starch fed group and levelled off to NG starch fed group. The results of this study indicated that dietary G starch may cause metabolic stress of fingerling L. rohita, as might consequently lead to the decrease antioxidant abilities, depressed immunity and reduced resistance to A. hydrophila infection. Subsequently, exposure of elevated temperature (32 °C) for one week mitigate immunosuppressive and oxidative stress effect of dietary G starch.
Sen, Shib Sankar; Sukumaran, V; Giri, Sib Sankar; Park, Se Chang
Psidium guajava L. is a well-known traditional medicinal plant widely used in folk medicine. To explore the anti-inflammatory activity of the flavonoid fraction of guava leaf extract (FGLE), we investigated its ability to suppress the levels of inflammatory mediators elevated by lipopolysaccharide (LPS) in Labeo rohita head-kidney (HK) macrophages. HK macrophages of L. rohita were treated with LPS in the presence or absence of the FGLE. We examined the inhibitory effect of FGLE on LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production. The inhibitory effect of FGLE on nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were investigated by RT-PCR and western blot. The effect of FGLE on proinflammatory cytokines tumour necrosis factor alpha (TNF-α) or interleukin-1β (IL-1β) was also investigated by ELISA and RT-PCR. The phosphorylation of three mitogen activated protein kinases (MAPK) molecules ERK, JNK and p38 was analysed by western blot analysis. FGLE inhibited LPS-induced NO and PGE2 production. It also effectively inhibited TNF-α, IL-1β, IL-10, iNOS, and COX-2 production in a concentration-dependent manner. In addition, FGLE suppressed the mRNA expression levels of TNF-α and IL-1β in LPS-stimulated HK macrophages. RT-PCR and western blot analysis showed that FGLE decreased both the mRNA and protein expression levels of LPS-induced iNOS and COX-2 in HK macrophages. FGLE suppresses the phosphorylation of MAPK molecules in LPS-stimulated HK macrophages. FGLE also significantly inhibited LPS-induced NF-κB transcriptional activity. The molecular mechanism by which FGLE suppresses the expression of inflammatory mediators appears to involve the inhibition of NF-κB activation, through the suppression of LPS-induced IκB-α degradation. Together these results suggest that FGLE contains potential therapeutic agent(s), which regulate NF-κB activation, for the treatment of inflammatory conditions in L. rohita macrophages.
Jena, S D; Behera, M; Dandapat, J; Mohanty, N
Xenobiotics-mediated environmental stress is an important determining factor in the maintenance of fish health as fishes are frequently exposed to such components. Increasing evidences indicate that acute and chronic xenobiotic exposure modulates ROS production, suppresses immune response and increase the incidence of fish diseases. In the present context an attempt has been made to study the in vivo effect of different concentrations of CuSO(4) (0.5, 1.00 or 2.00 ppm) on lipid peroxidation (an index of oxidative stress) and non enzymatic antioxidant status (glutathione and Ascorbic acid), in the muscle of a widely consumed freshwater fish Labeo rohita. From the out come of this study it is concluded that comparatively low dose of copper (0.5 ppm) induce mild oxidative stress in the experimental fish with concomitant elevation of GSH and AsA content of the muscle. However, high concentration of CuSO(4) (2.00 ppm) in the ambient water leads to severe oxidative stress manifested in the form of LPX and morphoanatomical alteration.
Krishna Priya, K; Ramesh, M; Saravanan, M; Ponpandian, N
The fate and effect of nanomaterials in the environment has raised concern about their environmental risk to aquatic organisms. Silica nanoparticles (SiO2-NPs) find its uses in various fields and are inevitably released into the environment. However, the ecotoxicological effects of SiO2-NPs on the freshwater fish remain poorly understood. The aim of this study was to evaluate the effect of different concentrations (1, 5 and 25mgL(-1)) of SiO2-NPs on certain hematological, ionoregulatory and enzymological profiles of a freshwater teleost fish Labeo rohita. Hematological parameters such as hemoglobin (Hb), hematocrit (Hct), red blood cells (RBC), white blood cells (WBC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) values were altered in SiO2-NPs treated groups. Likewise, plasma electrolytes such as plasma sodium (Na(+)), potassium (K(+)) and chloride (Cl(-)) levels and Na(+)/K(+) ATPase activity in gill of SiO2-NPs treated groups were altered in all concentrations throughout the study period (96h). The alterations of these parameters were found to be dependent on dose and exposure period. The results of the present study indicate that the alterations of these parameters may relate to physiological stress system to SiO2-NPs toxicity and also demonstrate that manufactured metal oxide NPs in aquatic environment may affect the health condition of the aquatic organisms.
Patra, Swagat Kumar; Chakrapani, Vemulawada; Panda, Rudra Prasanna; Mohapatra, Chinmayee; Jayasankar, Pallipuram; Barman, Hirak Kumar
Because little is known about the function of Sox2 (Sry-related box-2) in teleosts, the objective of this study was to clone and characterize Sox2 complementary DNA (cDNA) from the testis of Indian major carp, Labeo rohita (rohu). The full-length cDNA contained an open reading frame of 936 nucleotides bearing the typical structural features. Phylogenetically, Sox2 of L rohita was most closely related to freshwater counterparts than marine water. The sequence information of cDNA and genomic DNA together revealed that the Sox2 gene is encoded by an uninterrupted exon. Furthermore, comparative mRNA expression profile in various organs including proliferating spermatogonial stem cells (SSCs) suggested about the participatory role of Sox2 during fish male germ cell development and maintenance of stem cells. In support, we have also provided evidence that Sox2 protein is indeed present in rohu SSCs by Western blot analysis. The evolutionarily conserved high-mobility group box domain indicated its possible involvement in common networking pathways for stem cell maintenance and pluripotency between mammals and nonmammals. Our findings could be the first step toward the use of Sox2 as a potential biomarker for proliferating SSCs and understanding the transcriptional regulatory network involved during male germ cell development and maintenance in fish species.
Patnaik, Siddhi; Mohanty, Mausumee; Bit, Amrita; Sahoo, Lakshman; Das, Sachidananda; Jayasankar, Pallipuram; Das, Paramananda
Activin receptor type IIA (ActRIIA), a transmembrane serine/threonine kinase receptor is an important regulator of physiological traits, viz., reproduction and body growth in vertebrates including teleosts. However, existing knowledge of its role in regulating fish physiology is limited. To address this, we have cloned and characterized the ActRIIA cDNA of Labeo rohita (rohu), an economically important fish species of the Indian subcontinent. Comparative expression profiling of the receptor gene at various reproductive and growth stages supports to its role in promoting oocyte maturation, spermatogenesis and skeletal muscle development via interaction with multiple ligands of transforming growth factor-β (TGF-β) family. The full-length cDNA of rohu ActRIIA was found to be of 1587bp length encoding 528 amino acids. The three-dimensional structure of the intracellular kinase domain of rohu ActRIIA has also been predicted. Phylogenetic relationship studies showed that the gene is evolutionarily conserved across the vertebrate lineage implicating that the functioning of the receptor is more or less similar in vertebrates. Taken together, these findings could be an initial step towards the use of ActRIIA as a potential candidate gene marker for understanding the complex regulatory mechanism of fish reproduction and growth.
Giri, Sib Sankar; Sen, Shib Sankar; Chi, Cheng; Kim, Hyoun Joong; Yun, Saekil; Park, Se Chang; Sukumaran, V
The present study aimed to evaluate the effects of Psidium guajava L. (guava) leaves on the growth and immune response of the fish species Labeo rohita and its susceptibility to Aeromonas hydrophila infection. Diets containing five different concentrations of guava leaves (0% [basal diet], 0.1% [G1], 0.5% [G2], 1% [G3], and 1.5% [G4]) were fed to fish (average weight: 11.1 g) for 60 days. Various growth and immune parameters were examined 60 days post-feeding. Fish were challenged with A. hydrophila at the end of the trial, and mortalities were recorded over 15 days post-infection. We found that growth parameters such as percent weight gain (657.61 ± 9.74) and specific growth rate (3.37 ± 0.021) were significantly higher in G2 group than in the control (P < 0.05). Among the immune parameters examined, lysozyme levels (79.5 ± 5.1 U mL(-1)), leukocyte phagocytic activity (52 ± 4.3%), and alternative complement pathway activity (ACP) (186.1 ± 8.3 U mL(-1)) were significantly high (P < 0.05) in G2 fed group; there was, however, no significant effect of guava leaves at any concentration on plasma IgM level. Of the cytokine-related genes examined, interleukin-1beta (IL-1β) and tumour necrosis factor-alpha (TNF-α) were up-regulated in the head-kidney, intestine, and hepatopancreas of fish fed experimental diets, and expression was significantly higher in G2 and G3 than in the control group. In contrast, gene expression of IL-10, transforming growth factor-beta (TGF-β), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and transcription factor nuclear factor-κB (NF-κB) were down-regulated in the treatment groups. Moreover, fish fed the G2 diet exhibited a significantly higher post-challenge survival rate (66.66%). Collectively, these results suggest that dietary supplementation with guava leaves (at 0.5% concentration) could promote growth performance and strengthen immunity of L. rohita. Guava leaves therefore represent a promising feed additive
Majhi, Rakesh Kumar; Kumar, Ashutosh; Yadav, Manoj; Swain, Nirlipta; Kumari, Shikha; Saha, Ashish; Pradhan, Avinash; Goswami, Luna; Saha, Somdatta; Samanta, Luna; Maity, Apratim; Nayak, Tapas Kumar; Chattopadhyay, Subhasis; Rajakuberan, Chitra; Kumar, Abhishek; Goswami, Chandan
Sperm cells exhibit extremely high sensitivity in response to slight changes in temperature, osmotic pressure and/or presence of various chemical stimuli. In most cases throughout the evolution, these physico-chemical stimuli trigger Ca (2+)-signaling and subsequently alter structure, cellular function, motility and survival of the sperm cells. Few reports have recently demonstrated the presence of Transient Receptor Potential (TRP) channels in the sperm cells from higher eukaryotes, mainly from higher mammals. In this work, we have explored if the sperm cells from lower vertebrates can also have thermo-sensitive TRP channels. In this paper, we demonstrate the endogenous presence of one specific thermo-sensitive ion channel, namely Transient Receptor Potential Vanilloid family member sub type 1 (TRPV1) in the sperm cells collected from fresh water teleost fish, Labeo rohita. By using western blot analysis, fluorescence assisted cell sorting (FACS) and confocal microscopy; we confirm the presence of this non-selective cation channel. Activation of TRPV1 by an endogenous activator NADA significantly increases the quality as well as the duration of fish sperm movement. The sperm cell specific expression of TRPV1 matches well with our in silico sequence analysis. The results demonstrate that TRPV1 gene is conserved in various fishes, ranging from 1-3 in copy number, and it originated by fish-specific duplication events within the last 320 million years (MY). To the best of our knowledge, this is the first report demonstrating the presence of any thermo-sensitive TRP channels in the sperm cells of early vertebrates as well as of aquatic animals, which undergo external fertilization in fresh water. This observation may have implications in the aquaculture, breeding of several fresh water and marine fish species and cryopreservation of fish sperms.
Biswas, Saikat P; Jadhao, Arun G; Bhoyar, Rahul C; Palande, Nikhil V; Sinh, Dharmendra P
Nitric oxide (NO) is a chemically diffusible molecular messenger playing various roles in both vertebrates and invertebrates. Nitric oxide synthase (NOS) is the key enzyme in synthesis of NO. The neuroanatomical distribution pattern of neuronal nitric oxide synthase (nNOS) was studied and developing stages of Labeo rohita such as hatchlings (10-15mm), frys (15-35mm), semi-fingerlings (35-65mm), fingerlings (65-100mm) and adults (350-370mm) were used. In the telencephalon, nitrergic cells were observed in both pallial and subpallial regions along with entopeduncular nucleus suggesting the involvement of NO in the control of sensory functions throughout the development. In the diencephalon, nNOS positive neurons were localized in the nucleus preopticus periventricularis and preopticus parvocellularis throughout development while nucleus preopticus magnocellularis was found immunopositive only in adult specimens who suggest the involvement of NO in the hormonal regulation. nNOS immunoreaction was also noted in suprachaismatic nucleus, habenula, lateral tuberal nucleus, paraventricular organ and anterior division of preglomerular nucleus throughout development. In the mesencephalic region, nNOS immunoreactivity was seen in the optic tectum, torus longitudinalis, nucleus of median longitudinal fascicle and occulomotor nucleus indicate the role of NO in integration of visual inputs and modulates motor control of the eyes and movements. Caudally, in the rhombencephalon, the cerebellum, the nucleus reticularis, the octaval nucleus and the motor nucleus of vagal nerve were nNOS positive during development. nNOS reactive cells and fibers were noted in the spinal motor column, thus suggesting a role of NO in gestation and startle response from early development.
Ciji, A; Sahu, N P; Pal, A K; Akhtar, M S
An experiment was conducted to study the effect of sub-lethal nitrite exposure on sex steroids (testosterone and estradiol), cortisol and thyroid hormones (T3 and T4) of Labeo rohita juveniles. Fishes previously fed with normal or elevated levels of vitamin E (VE) and tryptophan for 60 days were exposed to sub-lethal nitrite for another 45 days with same feeding regime. There were nine treatment groups, viz. VE0TRP0-N, VE0TRP0+N, VE100TRP0-N, VE100TRP0+N, VE100TRP0.75+N, VE100TRP1.5+N, VE150TRP0+N, VE300TRP0+N and VE200TRP1+N. Except the groups VE0TRP0-N and VE100TRP0-N, all other groups were exposed to nitrite. At the end of the 45 days of nitrite exposure, serum samples were assayed for sex steroids, cortisol and thyroid hormones. The serum T3 and T4 levels decreased to the extent of 84.5 and 94.06%, respectively, upon nitrite exposure. Dietary supplementation with additional amounts of VE and tryptophan appears to reduce the decline of the production of T4. The serum testosterone and estradiol decreased 97.31 and 92.86%, respectively, upon nitrite exposure. Supplementation with additional amounts of VE was found to reverse nitrite-induced inhibition of testosterone and estradiol production. Serum cortisol increased upon nitrite exposure and unexposed (VE100-N) group showed lower levels, which were comparable to groups fed with elevated levels of VE. The overall results of the present study revealed that environmental nitrites have a negative impact on steroidogenesis, which can be overcome by dietary supplementation of elevated amounts of VE (minimum of 150 mg VE Kg diet(-1)) and to a lesser extent by tryptophan (only at the level of 1.5% of the diet).
Binukumari, S; Devi, K Anusiya; Vasanthi, J
Pesticides are widely used in modern agriculture to aid in the production of high quality food. However, some pesticides have the potential to cause serious health and environmental damage. Repeated exposure to sub-lethal doses of pesticides can cause physiological and behavioral changes in fish that reduce populations such as abandonment of nests and broods, decreased immunity to disease and increased failure to avoid predators. Monocrotophos is one of the organophosphorus pesticide used in this study. The median lethal concentration (LC50) of Monocrotophos to fish L. rohita for 96h was found to be 45.1ppm. In sublethal concentration (1/10th of LC50 96h value, 4.51ppm) fishes were exposed for 24, 48, 72, 96h and 10, 20 and 30days. Organs of fishes were sacrificed and tested for biochemical analysis. A significant decrease in protein, carbohydrate and lipids were observed throughout the study period when compared to the control. It is essential for assessing the ecological risk of these pesticides.
Prusty, Ashisa K.; PaniPrasad, Kurchetti; Mohanta, Kedar N.
Probiotics play an important role in growth increment, immune enhancement and stress mitigation in fish. Increasing temperature is a major concern in present aquaculture practices as it markedly deteriorates the health condition and reduces the growth in fish. In order to explore the possibilities of using probiotics as a counter measure for temperature associated problems, a 30 days feeding trial was conducted to study the hemato-immunological and apoptosis response of Labeo rohita (8.3±0.4 g) reared at different water temperatures, fed with or without dietary supplementation of a probiotic mixture (PM) consisting of Bacillus subtilis, Lactococcus lactis and Saccharomyces cerevisiae) (1011 cfu kg−1). Three hundred and sixty fish were randomly distributed into eight treatment groups in triplicates, namely, T1(28°C+BF(Basal feed)+PM), T2(31°C+BF+PM), T3(34°C+BF+PM), T4(37°C+BF+PM), T5(28°C+BF), T6(31°C+BF), T7(34°C+BF) and T8(37°C+BF). A significant increase (P<0.01) in weight gain percentage was observed in the probiotic fed fish even when reared at higher water temperature (34–37°C). Respiratory burst assay, blood glucose, erythrocyte count, total serum protein, albumin, alkaline phosphatase and acid phosphatase were significantly higher (P<0.01) in the probiotic fed groups compared to the non-probiotic fed groups. A significant (P<0.01) effect of rearing temperature and dietary probiotic mixture on serum myeloperoxidase activity, HSP70 level and immunoglobulin production was observed. Degree of apoptosis in different tissues was also significantly reduced in probiotic-supplemented groups. Hence, the present results show that a dietary PM could be beneficial in enhancing the immune status of the fish and also help in combating the stress caused to the organism by higher rearing water temperature. PMID:24979660
Yengkokpam, Sona; Debnath, Dipesh; Sahu, N P; Pal, A K; Jain, K K; Baruah, Kartik
Present experiment was conducted to study the effect of dietary protein levels on growth, immunity and anti-oxidative status of Labeo rohita fingerlings during feed deprivation followed by refeeding. Fish (5.44 ± 0.10 g) were deprived of feed for 3 weeks and then re-fed to satiation for 5 weeks with one of the diets containing 25 (25P), 30 (30P), 35 (35P) or 40 (40P) percent crude protein (CP) level. In addition to these groups, a control group (C) was also maintained by feeding to satiation level twice daily with a diet containing 30% CP throughout the experimental period. At the end of 8-weeks' trial, fish were challenged with Aeromonas hydrophila and survival was recorded for the next 7 days. Complete recovery of growth in terms of weight gain percentage was achieved in the fish fed 35 and 40% protein during refeeding. The body indices (condition factor and hepatosomatic index), haematological parameters and serum protein contents at the end of the experimental trial were not significantly different (P > 0.05) among different groups suggesting that the overall health of the fish was not compromised. However, respiratory burst activity and serum lysozyme activity were indicative of a better immune function in the higher protein fed groups (35P and 40P) than the lower protein groups (25P and 30P). Following challenge with Aeromonas hydrophila, survival rate, blood monocyte%, respiratory burst activity, serum lysozyme activity, serum protein and globulin were significantly higher (P < 0.05) in the 35P and 40P groups compared to the other groups. Further, fish fed lower dietary protein were not able to restore the activities of anti-oxidative enzymes (superoxide dismutase and catalase) in the liver. Conclusively, an improved disease resistance capability and immune status was observed in the fish fed a higher dietary protein (35-40%), even out-performing the daily-fed fish.
Sahoo, Bikash Ranjan; Basu, Madhubanti; Swain, Banikalyan; Dikhit, Manas Ranjan; Jayasankar, Pallipuram; Samanta, Mrinal
Toll-like receptors (TLRs) play key roles in sensing wide array of microbial signatures and induction of innate immunity. TLR2 in fish resembles higher eukaryotes by sensing peptidoglycan (PGN) and lipoteichoic acid (LTA) of bacterial cell wall and zymosan of yeasts. However, in fish TLR2, no study yet describes the ligand binding motifs in the leucine rich repeat regions (LRRs) of the extracellular domain (ECD) and important amino acids in TLR2-TIR (toll/interleukin-1 receptor) domain that could be engaged in transmitting downstream signaling. We predicted these in a commercially important freshwater fish species rohu (Labeo rohita) by constructing 3D models of TLR2-ECD, TLR2-TIR, and MyD88-TIR by comparative modeling followed by 40 ns (nanosecond) molecular dynamics simulation (MDS) for TLR2-ECD and 20 ns MDS for TLR2-TIR and MyD88-TIR. Protein (TLR2-ECD)–ligands (PGN, LTA, and zymosan) docking in rohu by AutoDock4.0, FlexX2.1, and GOLD4.1 anticipated LRR16–19, LRR12–14, and LRR20-CT as the most important ligand binding motifs. Protein (TLR2-TIR)—protein (MyD88-TIR) interaction by HADDOCK and ZDOCK predicted BB loop, αB-helix, αC-helix, and CD loop in TLR2-TIR and BB loop, αB-helix, and CD loop in MyD88-TIR as the critical binding domains. This study provides ligands recognition and downstream signaling. PMID:23956969
Siddiquee, M M R; Rahman, M F; Jahan, N; Jalal, K C A; Amin, S M N; Arshad, A
The fingerlings of indigenous carps such as catla (Catla catla), rohu (Labeo rohita) and mrigal (Cirrhinus mrigala) with exotic carps such as silver carp (Hypophthalmichthys molitrix), bighead carp (Aristichthys nobilis) and mirror carp (Cyprinus carpio) were cultured together in a fish pond at Bangladesh Agricultural University, Mymensingh, in order to determine the food electivity, dietary overlap and food competition among indigenous major carps and exotic carps. Phytoplankton (Chlorophyceae, Cyanophyceae), zooplankton (rotifers) were the dominant groups in the cultured pond. Chlorophyceae was dominant in the diet of rohu. Chlorophyceae and rotifers were the preferred food of catla. Mrigal preferred phytoplankton than zooplankton. Rohu showed positive electivity for zooplankton. Silver carp consumed large quantity of phytoplankton and also preferred rotifers. Chlorophyceae was the dominant food group in the diet of bighead. Mirror carp also preferred plant food organisms dominated by Chlorophyceae. Bighead had positive trends towards phytoplankton. Both mrigal and mirror carp had positive electivity towards phytoplankton. The higher level of dietary overlap occurred between rohu and silver carp followed by between rohu and bighead carp and between catla and silver carp. The lowest level of dietary overlaps occurred between rohu and mirror carp.
Giri, Sib Sankar; Sen, Shib Sankar; Jun, Jin Woo; Sukumaran, V.; Park, Se Chang
Multifarious applications of Bacillus licheniformis VS16-derived biosurfactant were explored. Labeo rohita fingerlings were injected intraperitoneally with 0.1 mL of phosphate-buffered saline (PBS) containing purified biosurfactant at 0 (control), 55 (S55), 110 (S110), 220 (S220), or 330 (S330) μg mL-1 concentrations. Various immunological parameters and the expression of immune-related genes were measured at 7, 14, and 21 days post-administration (dpa). At 21 dpa, fish were challenged with Aeromonas hydrophila and mortality was recorded for 14 days. Immune parameters such as lysozyme levels (39.29 ± 2.14 U mL-1), alternative complement pathway (61.21 ± 2.38 U mL-1), and phagocytic activities (33.37 ± 1.2%) were maximum (P < 0.05) in the S220 group at 14 dpa; but immunoglobulin levels (11.07 ± 0.83 mg mL-1) were highest in the S220 group at 7 dpa, compared to that in controls. Activities of digestive enzymes (amylase, protease, and lipase) were higher (P < 0.05) in the S220 and S330 groups than in the control group. Regarding cytokine gene expression, pro-inflammatory cytokines (TNF-α and IL-1β) were down-regulated (P < 0.05) in the S220 and S330 groups. Expression of IL-10, TGF-β, and IKB-α were up-regulated in the S220 and S330 groups at 14 dpa, with the highest levels in the S220 group. The expression of NF-κB p65 and IKK-β were down-regulated in treatment groups, and were lowest (P < 0.05) in the S220 group. The highest post-challenge survival rate (72.7%) was recorded in S220 group. Further, the potential of this substance to inhibit biofilm formation, and heavy metal removal from vegetables were also evaluated. Biosurfactant was effective in inhibiting biofilm formation up to 54.71 ± 1.27%. Moreover, it efficiently removed cadmium (Cd) from tested vegetables such as carrot, radish, ginger, and potato, with the highest removal efficiency (60.98 ± 1.29%) recorded in ginger contaminated with Cd. Collectively, these results suggest that isolated
Chakrapani, Vemulawada; Patra, Swagat Kumar; Panda, Rudra Prasanna; Rasal, Kiran Dashrath; Jayasankar, Pallipuram; Barman, Hirak Kumar
Recent advances in gene editing techniques have not been exploited in farmed fishes. We established a gene targeting technique, using the CRISPR/Cas9 system in Labeo rohita, a farmed carp (known as rohu). We demonstrated that donor DNA was integrated via homologous recombination (HR) at the site of targeted double-stranded nicks created by CRISPR/Cas9 nuclease. This resulted in the successful disruption of rohu Toll-like receptor 22 (TLR22) gene, involved in innate immunity and exclusively present in teleost fishes and amphibians. The null mutant, thus, generated lacked TLR22 mRNA expression. Altogether, this is the first evidence that the CRISPR/Cas9 system is a highly efficient tool for targeted gene disruption via HR in teleosts for generating model large-bodied farmed fishes.
Hemaprasanth; Singh, Ravinder; Raghavendra, A; Sridhar, N; Raghunath, M R; Eknath, A E
Study was conducted to find out the comparative susceptibility of fingerlings of seven species of carps (Labeo fimbriatus, L. rohita, L. calbasu, Catla catla, Ctenopharyngodon idella, Cyprinus carpio and Hypophthamichthys molitrix) grown under both mono and polyculture to Lernaea cyprinacea infection. Under monoculture, C. carpio, L. rohita and L. calbasu, did not acquire Lernaea infection and were thus considered resistant, whereas C. idella, H. molitrix, C. catla and L. fimbriatus were susceptible. Even challenge with higher infective doses of copepodids under monoculture did not result in infection in the resistant fish species. The resistance of L. rohita and C. carpio to Lernaea infection under monoculture was not sustained when these two fish species were maintained in polyculture along with susceptible fish species. Labeo calbasu, even under polyculture, however, did not acquire Lernaea infection indicating that this fish species is the most resistant and least preferred host for this parasite. Similarly, C. carpio, L. rohita and L. calbasu when grown together in polyculture and exposed to a higher infective dose (120 copepodids/fish) also did not develop the infection. The possible reasons for differences in susceptibility shown by these carp species in monoculture and the loss of resistance by rohu and common carp while in polyculture with susceptible species are discussed. The ability of resistant fish species to prevent establishment of anchor worms on them under monoculture can be utilized to control this parasitic infection commonly encountered in culture ponds.
Majumdar, Ranendra K; Dhar, Bahni; Roy, Deepayan; Saha, Apurba
'Kalia', a popular preparation of Rohu fish, packed in four-layered laminated retort pouch was processed in a steam/air mixture over-pressure retort at 121.1 °C to three different F 0 values of 7, 8 and 9 min. Time-temperature data were collected during heat processing using an Ellab Sterilization Monitoring System. Texture profile such as hardness, springiness, gumminess and chewiness decreased as the F 0 value increased. The L* values decreased whereas a* and b* values increased with increasing F 0 value. Based on the commercial sterility, sensory evaluation, colour and texture profile analysis, F 0 value of 8 min and cook value of 66 min, with a total process time of 41.7 min at 121.1 °C was found satisfactory for the preparation of Rohu fish curry (Kalia) in retort pouches.
Van Steenberge, M; Gajdzik, L; Chilala, A; Snoeks, J; Vreven, E
Labeo rosae, a species with a native range in Southern Africa, was discovered in the Congo basin by re-identification of two museum specimens previously identified as Labeo mesops. The occurrence of this species in the upper Congo implies a range extension of the species of more than 1000 km. Although the species' distribution is mirrored by that of some other Cypriniformes, its occurrence in the Congo might be due to introduction by humans.
Rangacharyulu, P V; Giri, S S; Paul, B N; Yashoda, K P; Rao, R Jagannatha; Mahendrakar, N S; Mohanty, S N; Mukhopadhyay, P K
Fermented silkworm pupae (SWP) silage or untreated fresh SWP pastes were incorporated in carp feed formulations replacing fishmeal. The feed formulations were isonitrogenous (30.2-30.9% protein) and isocaloric (ME = 2905-2935 kcal/kg). Feeding under a polyculture system consisting of 30% each of catla (Catla catla), mrigal (Cirrhinus mrigala) and rohu (Labeo rohita) with 10% silver carps (Hypophthalmychthys molitrix) was carried out in ponds to evaluate the nutritive quality of SWP silage. Survival rate, feed conversion ratio and specific growth rate, respectively, were 84.2%, 2.10 and 2.39 for fermented SWP silage, 65.8%, 2.98 and 2.26 for untreated SWP and 67.5%, 3.16 and 2.20 for fishmeal indicating clearly that the fermented SWP silage was nutritionally superior to untreated SWP or fishmeal. The dietary influence on the proximate composition of whole fish was marginal.
Verma, Dev Kumar; Rathore, Gaurav; Pradhan, Pravata Kumar; Sood, Neeraj; Punia, Peyush
Filamentous bacteria overlaying ulcerated area on the body surface were observed in the wet-mout preparation from a moribund goldfish with saddle back appearance. The causative agent was identified as Flavobacterium columnrae, on the basis of biochemical test, species-specific polymerase chain reaction (PCR) and sequencing of 16S rDNA gene with the universal bacterial primers. Furthermore, the strain (ING-1) attributed to genomovar II in 16S rDNA PCR-restriction fragment length polymorphism (PCR-RFLP) and sequence analysis. In phylogenetic analysis, the strain ING-1, produced typical columnaris disease symptoms in rohu (Labeo rohita) fingerlings within 10 days. This is a new record about molecular detection and identification of Flavobacterium columnare, occurring naturally on a new host Carassius auratusin India.
Molecular cloning and characterization of LrTLR4, analysis of its inductive expression and associated down-stream signaling molecules following lipopolysaccharide stimulation and Gram-negative bacterial infection.
Samanta, Mrinal; Basu, Madhubanti; Swain, Banikalyan; Paichha, Mahismita; Lenka, Saswati S; Das, Surajit; Jayasankar, Pallipuram; Maiti, Nikhil Kumar
Toll-like receptors (TLRs) play key roles in innate immunity from lower to higher vertebrates. Among various TLR types, TLR4 was reported to recognize LPS in higher vertebrates resulting in the activation of down-stream signaling pathway. Except in some teleosts, function of TLR4 in most fish species including rohu (Labeo rohita) a commercially important fish species in the South-East Asian countries remained unknown. To investigate it, full-length cDNA of Labeo rohita TLR4 (LrTLR4) was cloned, and it consisted of 2729 bp, with a single ORF of 2469 bp encoding a polypeptide of 822 aa with a predicted molecular mass of 94.753 kDa. Structurally, LrTLR4 consisted of 25 LRRs (leucine rich repeat regions), one TM (trans-membrane) domain and one TIR (Toll/interleukin-1 receptor) domain, and was similar to higher vertebrate's TLR4. Phylogenetically, LrTLR4 exhibited highest (85%) identity with the common carp TLR4b amino acids sequence, and formed a separate subgroup in the phylogenetic tree. LrTLR4 was widely expressed in all tested organs/tissues, and amidst the tissues highest expression was detected in blood and the lowest in eye. In response to LPS-stimulation, LrTLR4 was induced with the activation of MyD88-dependent and TRIF-dependent signaling pathway resulting in pro-inflammatory cytokines (interleukin 6 and 8) and type I IFN gene expression. Infection of rohu with a Gram-negative fish pathogen (Aeromonas hydrophila), also activated LrTLR4. Together, these findings suggest the important role of TLR4 in LPS sensing and augmentation of innate immunity against Gram-negative bacterial infection in fish.
Dang, Son Thi Thanh; Dalsgaard, Anders
Integrated livestock-fish aquaculture utilizes animal excreta and urine as pond fertilizers to enhance growth of plankton and other microorganisms eaten by the fish. In Vietnam, pigs are commonly integrated with fish and horticulture in household-based VAC systems (vuon = garden; ao = pond; chuong = pigsty), but the level of fecal contamination in the fish produced is unknown. This study was carried out to assess the level Escherichia coli contamination of fish meat and gut contents of grass carp (Ctenopharyngodon idellus), silver carp (Hypophthalmichthys molitrix), and rohu (Labeo rohita) cultured in randomly selected five VAC ponds (with pig manure) and five non-VAC ponds (without pig manure) at sites in periurban Hanoi, Vietnam. Fish muscle tissue samples contained E. coli at <10 or 320 or 820 CFU/g, regardless of the culture system from which they originated. In contrast, the intestinal contents of fish raised in manure-fed ponds contained E. coli at 4.75, 5.25, and 5.07 log CFU/g for silver carp, grass carp, and rohu, respectively, about 100 times higher than the contamination of fish from the control ponds. The results indicate that muscle tissue of fish raised in VAC systems has a low level of fecal contamination despite high levels of E. coli in their gut. Thus, a critical point to control food safety of such fish is the prevention of fecal cross-contamination during degutting and cleaning of the fish at markets and in the home.
Behera, Bijay Kumar; Kumari, Kavita; Baisvar, Vishwamitra Singh; Rout, Ajaya Kumar; Pakrashi, Sudip; Paria, Prasenjet; Jena, J K
In the present study, the complete mitochondrial genome sequence of Labeo gonius is reported using PGM sequencer (Ion Torrent). The complete mitogenome of L. gonius is obtained by the de novo sequences assembly of genomic reads using the Torrent Mapping Alignment Program (TMAP) which is 16 614 bp in length. The mitogenome of L. gonius comprised of 13 protein-coding genes, 22 tRNAs, 2 rRNA genes, and D-loop as control region along with gene order and organization, being similar to most of other fish mitogenomes of NCBI databases. The mitogenome in the present study has 99% similarity to the complete mitogenome sequence of Labeo fimbriatus, as reported earlier. The phylogenetic analysis of Cypriniformes depicted that their mitogenomes are closely related to each other. The complete mitogenome sequence of L. gonius would be helpful in understanding the population genetics, phylogenetics, and evolution of Indian Carps.
Hossain, M Motaleb; Rahman, Mahabubur; Hassan, M Nazmul; Nowsad, A A K M
Post-harvest loss of catla (Catla catla), rohu (Labeo rohita), mrigal (Cirrhinus mrigala), silver carp (Hypophthalmichthys molitrix), grass carp (Ctenopharyngodon idella) and sarpunti (Puntius sarana) in a single distribution chain from harvest in Mymensingh to retail sale Rangpur town were determined, in order to obtain information on quality deterioration and existing handling and icing conditions so that suggestions for improving such practices can be made. Quality defect points of the fish at different steps of distribution channels were determined using a sensory based quality assessment tool. Percent quality loss of fish at each step of distribution was calculated from the number of cases that crossed sensory quality cut-off points. Neither of the fish lost their quality when they were in the farm gate, during transportation and in wholesale markets in Rangpur but most of the fishes lost their quality at the retail fish shops. The quality loss was 8, 12, 8, 6, 10 and 14% in case of C. catla, C. mrigala, L. rohita, H. molitrix, C. idella and P. sarana respectively in the retail markets. Fishes were not properly handled, bamboo baskets wrapped with polythene sheet were used as carrying container and inadequate ice was used during transportation. Retailers were found to be more proactive in the use of ice. However, most of the fishes were deteriorated during retail sale. The losses of farmed fishes could be minimized by adopting good handling practices like using insulated container and adequate icing.
Effects of dietary administration of stinging nettle (Urtica dioica) on the growth performance, biochemical, hematological and immunological parameters in juvenile and adult Victoria Labeo (Labeo victorianus) challenged with Aeromonas hydrophila.
Ngugi, Charles C; Oyoo-Okoth, Elijah; Mugo-Bundi, James; Orina, Paul Sagwe; Chemoiwa, Emily Jepyegon; Aloo, Peninah A
We investigated effects of dietary administration of stinging nettle (Urtica dioica) on growth performance, biochemical, hematological and immunological parameters in juvenile and adult Victoria Labeo (Labeo victorianus) against Aeromonas hydrophila. Fish were divided into 4 groups and fed for 4 and 16 weeks with 0%, 1%, 2% and 5% of U. dioica incorporated into the diet. Use of U. dioica in the diet resulted in improved biochemical, hematological and immunological parameters. Among the biochemical parameters; plasma cortisol, glucose, triglyceride and cholesterol decreased while total protein and albumin in fish increased with increasing dietary inclusion of U. dioica. Among the haematology parameters: red blood cell (RBC), white blood cell (WBC) counts, haematocrit (Htc), mean cell haemoglobin (MCH), mean cell haemoglobin concentration (MCHC) and netrophiles increased with increasing dietary inclusion levels of U. dioica, some depending on the fish age. Serum immunoglobulins, lysozyme activity and respiratory burst were the main immunological parameters in the adult and juvenile L. victorianus measured and they all increased with increasing herbal inclusion of U. dioica in the diet. Dietary incorporation of U. dioica at 5% showed significantly higher relative percentage survival (up to 95%) against A. hydrophila. The current results demonstrate that using U. dioica can stimulate fish immunity and make L. victorianus more resistant to bacterial infection (A. hydrophila).
Wang, Jian; Guo, Huarong; Zhang, Shicui; Yin, Licheng; Guo, Bin; Wang, Shaojie
A full-length cDNA encoding translationally controlled tumor protein of marine flatfish turbot ( Scophthalmus maximus), SmTCTP, was isolated with rapid amplification of cDNA Ends (RACE). SmTCTP consisted of a 5' untranslated region (UTR) of 84 bp, a 3' UTR of 451 bp and an open reading frame (ORF) of 513 bp, encoding a protein of 170 amino acid residues, which contained two signature sequences of TCTP family. The 5'UTR of SmTCTP started with a 5'-terminal oligopyrimidine tract (5'-TOP), a typical feature for translationally controlled mRNAs. The deduced amino acid sequence of SmTCTP was similar to the other known vertebrate TCTPs in a range of 58.8% to 64.1%. The length of fish TCTPs was diverse among species, e.g., TCP of turbot and sea perch ( Lateolabrax japonicus) is 170 aa in length, while that of zebrafish ( Danio rerio) and rohu ( Labeo rohita) is 171 aa in length. Northern blot analysis revealed that SmTCTP has only one type of mRNA. Its expression level in albino skin was slightly higher than that in normal skin. We constructed the pET30a- SmTCTP expression plasmid. The recombinant protein of His-tag SmTCTP was over-expressed in E. coli, purified and identified with peptide mass fingerprinting. These results may pave the way of further investigation of the biological function of TCTP in fish.
Kant, Rajni; Haq, S; Srivastava, H C; Sharma, V P
Mosquito control with the use of insecticides is faced with the challenges of insecticide resistance in disease vectors, community refusal, their high cost, operational difficulties, and environmental concern. In view of this, integrated vector control strategies with the use of larvivorous fishes such as Guppy (Poecilia reticulata) and Gambusia (G. affinis) as biological control agents were used in controlling mosquito breeding in different types of breeding places such as intradomestic containers, various types of wells, rice-fields, pools, ponds and elsewhere in malaria prone rural areas of central Gujarat. Attempts were also made to demonstrate composite fish culture in unused abandoned village ponds by culturing Guppy along with the food fishes such as Rohu (Labeo rohita), Catla (Catla catla) and Mrigal (Cirrhinus mrigala). Income generated from these ponds through sale of fishes was utilized for mosquito control and village development. The technology was later adopted by the villagers themselves and food fish culture was practised in 23 ponds which generated an income of Rs 1,02,50,992 between 1985 and 2008. The number of villages increased from 13 to 23 in 2008 and there was also gradual increase of income from Rs 3,66,245 in 1985-90 to Rs 55,06,127 in 2002-08 block. It is concluded that larvivorous fishes can be useful tool in controlling mosquito breeding in certain situations and their use along with composite fish culture may also generate income to make the programme self-sustainable.
Nahiduzzaman, Md; Hassan, Md Mahbubul; Roy, Pankoz Kumar; Hossain, Md Akhtar; Hossain, Mostafa Ali Reza; Tiersch, Terrence R
A sperm cryopreservation protocol for the Indian major carp, Labeo calbasu, was developed for long-term preservation and artificial fertilization. Milt collected from mature male fish were placed in Alsever's solution (296mOsmolkg(-1)) to immobilize the sperm. Cryoprotectant toxicity was evaluated by motility assessment with dimethyl sulfoxide (DMSO) and methanol at 5, 10 and 15% concentrations. DMSO was more toxic at higher concentrations than methanol, and consequently 15% DMSO was excluded from further study. A one-step cooling protocol (from 5 to 80°C) with two cooling rates (5 and 10°C/min) was carried out in a computer-controlled freezer (FREEZE CONTROL(®) CL-3300; Australia). Based on post-thaw motility, the 10°C/min cooling rate with either 10% DMSO or 10% methanol yielded significantly higher (P=0.011) post-thaw motility than the other rate and cryoprotectant concentrations. Sperm thawed at 40°C for 15s and fresh sperm were used to fertilize freshly collected L. calbasu eggs and significant differences were observed (P=0.001) in percent fertilization between cryopreserved and fresh sperm as well as among different sperm-to-egg ratios (P=0.001). The highest fertilization and hatching rates were observed for thawed sperm at a sperm-to-egg ratio of 4.1×10(5):1. The cryopreservation protocol developed can facilitate hatchery operations and long-term conservation of genetic resources of L. calbasu.
Rajesh, Sandya Chinna; Banerjee, Sayani; Patra, Avijit; Dash, Gadadhar; Abraham, Thangapalam Jawahar
As new pathogenic strains are emerging and threatening aquaculture development, myxosporeans (Myxozoa) are receiving much attention in recent years. Myxosporean taxonomy is traditionally based on morphology of the myxospore stage. Molecular data on Indian myxosporeans are rare. In this report, the 18S rRNA gene sequence of Myxobolus cuttacki infecting gill lamellae of minor carp Labeo bata (Ham.) and its phylogenetic relationship with other myxobolids are described for the first time. The plasmodia of M. cuttacki were 0.5-0.9 mm in size and whitish with a round to oval shape. The mean mature spore size was 16.10×7.05 μm. The 18S rRNA nucleotide sequence with 1703 bp of M. cuttacki (accession number KF465682) clustered phylogenetically with other Myxobolus spp. infecting cyprinid gills with 78-90% homogeneity. The gill lamellae infecting M. catmrigalae (KC933944) and M. orissae (KF448527) of Indian major carp Cirrhinus mrigala from India, exhibited 86% and 81% homogeneity with M. cuttacki, respectively. The infection rate was low to moderate on the gills which can have a negative impact on respiratory and physiological functions and subsequently on fish production. PMID:27843987
Abdel-Ghaffar, Fathy; Morsy, Kareem; Bashtar, Abdel-Rahman; El-Ganainy, Sahar; Gamal, Shams
In the present study, the morphology and morphometric characterization of Thelohanellus niloticus sp. nov., a new myxozoan belonging to genus Thelohanellus Kudo, 1933 (Myxosporea, Bivalvulida) infecting the gills of Labeo niloticus (Osteichthyes, Cyprinidae), were described for the first time from the River Nile at El-Minia Governorate, Egypt. Forty-one out of 78 (52.6 %) of the examined fish were infected. The infection was observed as irregular, milky whitish, cyst-like plasmodia (up to 0.8 mm in diameter) attached to the gill filaments of the host fish. These plasmodia contained tear-shaped myxospores with slightly tapering anterior and rounded posterior ends. Each spore has a single pyriform polar capsule. Spores measured about 23.3 ± 0.3 (20.4-27.1) μm long and 13.4 ± 0.4 (11.5-14.2) μm wide. The polar capsule was 11.7 ± 0.3 (9.2-12.5) μm long and 4.7 ± 0.3 (3.5-6.2) μm wide, containing a polar filament coiled perpendicular to the longitudinal axis of the spore body making eight turns. Occasionally, an oblong, irregular-shaped mass of protoplasm with a slightly oval nucleus (1.4 μm in diameter) and a small iodinophilous vacuole measured 0.85 ± 0.2 μm (0.73-1.2 μm) were observed in the spore. Due to the lack of the second polar capsule characterizing Myxobolus sp., the present parasite is placed within the genus Thelohanellus. Based on morphological differences (compared with other members of Thelohanellus Kudo, 1933) and the host specificity, this species is described as a new one of the genus Thelohanellus recorded for the first time in Egypt.
Chaturvedi, Anshumala; Mohindra, Vindhya; Singh, Rajeev K; Lal, Kuldeep K; Punia, Peyush; Bhaskar, Ranjana; Mandal, Anup; Narain, Lalit; Lakra, W S
We examined population structure of Labeo dero (Hamilton, 1822) from different riverine locations in India using 10 polymorphic allozyme and eight microsatellite loci. For analysis, 591 different tissue samples were obtained from commercial catches covering a wide geographic range. Allozyme variability (An = 1.28-1.43, Ho = 0.029-0.071) was much lower than for microsatellites (An = 4.625-6.125, Ho = 0.538-0.633). Existence of rare alleles was found at three allozyme (MDH-2, GPI and PGDH) and at two microsatellite loci (R-3 and MFW-15). Deviation from Hardy-Weinberg equilibrium (P < 0.05, after the critical probability levels were adjusted for sequential Bonferroni adjustment) could be detected at three loci (EST-1, -2 and XDH) whereas, after correction for null alleles, two microsatellite loci (MFW-1,-15) deviated from HWE in the river Yamuna. Fst for all the samples combined over all allozyme loci was found to be 0.059 suggesting that 5.9% of the total variation was due to genetic differentiation while microsatellite analysis yielded 0.019 which was concordant to mean Rst (0.02). Hierarchical partition of genetic diversity (AMOVA) showed that greater variability (approx. 95%) was due to within population component than between geographical regions. Based on distribution of genetic differentiation detected by both markers, at least five different genetic stocks of L. dero across its natural distribution could be identified. These results are useful for the evaluation and conservation of L. dero in natural water bodies.
Das, Suchismita; Choudhury, Shamim Sultana
The aim of this study was to assess the regional impacts of heavy metals (Mn, Fe, Mg, Ca, Cu, Zn, Cd, Cr, Pb and Ni) on water, sediment and a native, teleost fish species, Labeo angra, inhabiting a flood plain wetland of Barak River in Assam, India. Heavy metal concentrations in the water, sediments and fish were measured; bioaccumulation factor, metal pollution index as well as condition indices were calculated, to assess the pollution load and health status of the fish. Multivariate statistical analysis was used on wetland water and sediment heavy metals to ascertain the possible sources and seasonal variations of the pollutants. Results showed that most heavy metals in the wetland water and sediments exceeded the water (drinking and irrigation) and sediment quality guidelines, respectively. Seasonal variations were observed for geogenic heavy metals, Mn, Fe, Mg and Ca while no seasonal variations were observed for anthropogenic heavy metals, Cu, Cd, Cr, Pb and Ni. Multivariate statistical analysis showed that there was strong correlation between geogenic and anthropogenic heavy metals in water and sediment, both originating from the common anthropogenic sources. Accumulation of most of the metals in all the tissues was above the safe limits as recommended by the Food and Agriculture Organization. High bioaccumulation factors and metal pollution index for these metals in the different tissues revealed that metals were extensively bio-accumulated and bioconcentrated. Condition indices in fish from the wetland suggested metabolic abnormalities.
Mataba, Gordian Rocky; Verhaert, Vera; Blust, Ronny; Bervoets, Lieven
The aim of the present study was to assess the distribution of trace elements in the aquatic ecosystem of the Thigithe river. Samples of surface water, sediment and fish were collected up- and downstream of the North Mara Gold Mine (Tanzania) and following trace elements were analysed: As, Cd, Co, Cr, Cu, Hg, Ni, Pb and Zn. Trace element concentrations in surface water were below or near the detection limit. Regarding the sediments, relative high concentrations of arsenic at all sites and high levels of mercury at a site downstream of the mine where artisanal mining is performed were observed. Trace element concentrations in Ningu fish tissues (Labeo victorianus) were comparable to slightly higher than levels in fishes from unpolluted environments. For none of the measured human health risk by consumption of fish from the Thigithe river is expected when the Tanzanian average amount of 17 g/day is consumed. However, for Hg and As the advised maximum daily consumption of Ningu fish was lower than 100g. As a result fishermen and people living along the shores of the river consuming more fish than the average Tanzanian fish consumption set by the FAO (2005) are possibly at risk.
Kaur, Harpreet; Singh, Ranjeet
A survey of parasites of freshwater fishes in Harike and Ropar Wetlands of Punjab (India) revealed the presence of two new myxosporean species belonging to the genus Myxobolus Butschli 1882. Spores of the first species, M. duodenalis sp. nov. parasitize the inner wall of the duodenum of Wallago attu (Bloch and Schneider) (Cypriniformis: Cyprinidae) vern. mulli are characterized by a broad, pyriform shape with a blunt anterior end and measures 9.0 × 3.20 μm. Spore valves are thin, smooth, and symmetrical with no parietal folds. There are two polar capsules, prominently unequal and pyriform in shape, with a narrow anterior end and a rounded posterior end. The larger polar capsule measures 4.70 × 1.76 μm and the smaller 2.76 × 1.06 μm. The larger polar capsule occupies more than half, while the smaller one occupies one third of the spore body cavity. An intercapsular process is absent. Spores of the second species, Myxobolus patialensis sp. nov. parasitize the caudal fin of Labeo rohita (Ham. 1822) (Cypriniformis: Cyprinidae) vern. rohu are pyriform in valvular view, measuring 11.28 × 6.67 μm. The two shell valves are asymmetrical and contain five parietal folds along the posterior end of the spore. Two anteriorly situated polar capsules, elongated and oval in shape are prominently unequal in size. The larger polar capsule measures 4.8 × 3.1 μm and smaller one 1.70 × 1.51 μm. The larger polar capsule lies obliquely to the spore axis and the smaller one is at the same level but pointing outward anteriolaterally. An intercapsular process is absent. This species is characterized by having a prominent ridge on the shell surface anteriolaterally on the side of the smaller polar capsule.
Hassan, M A; Aftabuddin, Md; Meena, D K; Mishal, P; Gupta, S Das
A 60-day feeding trial was conducted to evaluate the efficacy of distiller's grain soluble (brewery waste) as a prospective ingredient to substitute expensive and high demand feed component, soybean meal for farming Labeo bata in cages installed in tropical reservoir. Two isonitrogenous and isocaloric diets comprising brewery waste (49.2 % CP) as test diet and soybean meal (44.4 % CP) as reference diet were formulated and extruded to obtain 2-mm floating pellets. The efficacy of the diets was tested in terms of survival (%), live weight gain (%), SGR (%/day), FCR, PER and ANPU and recorded 65 ± 0.2, 96 ± 8.1, 1.9 ± 0.1, 2.5 ± 0.02, 1.4 ± 0.1, 20.3 ± 2.0 and 66 ± 0.6, 112 ± 9.8, 2.2 ± 0.1, 2.2 ± 0.2, 1.6 ± 0.1 and 20 ± 2.1, respectively, for soybean and brewer's waste-based formulated feed. The analyses of results revealed that survival, growth parameters and biochemical composition of whole body tissue did not differ significantly (p > 0.05) despite complete replacement of soybean meal by brewery waste. However, the cost estimate of diet revealed marked reduction of feed cost of Rs. 9.2/kg (33.8 %) in the test diet as compared to the reference diet. The study suggests that brewery waste could effectively replace soybean meal without effecting survival and growth of the fish. The finding thus may pave a productive way for reducing environmental pressure of disposal of an agro-industrial waste.
Goswami, Mukunda; Yadav, Kamalendra; Dubey, Akhilesh; Sharma, Bhagwati S; Konwar, Rituraj; Kumar, Ravindra; Nagpure, Naresh S; Lakra, Wazir S
Labeo rohita (rohu) is one of most important of Indian major carps, which is highly ranked among the important freshwater aquaculture species in the world. Heavy metals are major risk factors for aquatic health, which are biomagnified through the food chain. The present study was carried out to investigate the effect of different divalent salts of two heavy metals, such as zinc (ZnCl(2), ZnSO(4) and ZnNO(3)) and cadmium (CdCl(2) and CdSO(4)), in an established fish cell line, RF developed from fin tissue of L. rohita. The RF cell line was used for assessment of heavy metal cytotoxicity through various endpoint assays, including maximum tolerated dose (MTD) determination, 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, neutral red (NR) uptake assay, and Coomassie brilliant blue (CBB) assay. Results revealed that these heavy metal salts were cytotoxic to the RF cell line at varied concentrations. MTD values were found to be 1.563, 3.125, 6.25, 12.5 and 25 mg/L for CdCl(2), CdSO(4), ZnCl(2), Zn(NO(3))(2) and ZnSO(4), respectively. The half-maximal inhibitory concentration values calculated by MTT, NR and CB assay were 53.83 ± 7.02, 58.03 ± 9.12 and 79.20 ± 15.27 for ZnSO(4), 26.44 ± 7.01, 36.60 ± 7.82 and 155.6 ± 14.75 for Zn(NO(3))(2), 20.26 ± 17.95, 16.94 ± 7.05 and 87.54 ± 7.58 for ZnCl(2), 5.166 ± 0.57, 15 ± 1 and 41.80 ± 8.38 for CdSO(4) and 4.966 ± 0.56, 9.56 ± 1.73 and 20.93 ± 4.47 for CdCl(2). This study establishes the RF cell line as an in vitro tool for assessment and monitoring of heavy metal concentration in the aquatic environment.
Mohilal, Naorem; Hemananda, Thounaojam
Survey on Trichodinid ciliophorans from the fresh water fishes of Manipur revealed three known species of the genus Tripartiella from the gills of major carps Labeo rohita (Hamilton); Cirrhinus mrigala (Hamilton); Catla catla and Ciprinus carpio. These are redescribed in this communication.
Dubey, Saurabh; Avadhani, Kiran; Mutalik, Srinivas; Sivadasan, Sangeetha Madambithara; Maiti, Biswajit; Girisha, Shivani Kallappa; Venugopal, Moleyur Nagarajappa; Mutoloki, Stephen; Evensen, Øystein; Karunasagar, Indrani; Mweemba Munang′andu, Hetron
The use of oral vaccination in finfish has lagged behind injectable vaccines for a long time as oral vaccines fall short of injection vaccines in conferring protective immunity. Biodegradable polymeric nanoparticles (NPs) have shown potential to serve as antigen delivery systems for oral vaccines. In this study the recombinant outer membrane protein A (rOmpA) of Edwardsiella tarda was encapsulated in chitosan NPs (NP-rOmpA) and used for oral vaccination of Labeo fimbriatus. The rOmpA purity was 85%, nanodiameter <500 nm, encapsulation efficiency 60.6%, zeta potential +19.05 mV, and there was an in vitro release of 49% of encapsulated antigen within 48 h post incubation in phosphate-buffered saline. Empty NPs and a non-formulated, inactivated whole cell E. tarda (IWC-ET) vaccine were used as controls. Post-vaccination antibody levels were significantly (p = 0.0458) higher in the NP-rOmpA vaccinated fish (Mean OD450 = 2.430) than in fish vaccinated with inactivated whole cell E. tarda (IWC-ET) vaccine (Mean OD450 = 1.735), which corresponded with post-challenge survival proportions (PCSP) of 73.3% and 48.28% for the NP-rOmpA and IWC-ET groups, respectively. Serum samples from NP-rOmpA-vaccinated fish had a higher inhibition rate for E. tarda growth on tryptic soy agar (TSA) than the IWC-ET group. There was no significant difference (p = 0.989) in PCSPs between fish vaccinated with empty NPs and the unvaccinated control fish, while serum from both groups showed no detectable antibodies against E. tarda. Overall, these data show that the NP-rOmpA vaccine produced higher antibody levels and had superior protection over the IWC-ET vaccine, showing that encapsulating OmpA in chitosan NPs confer improved protection against E. tarda mortality in L. fimbriatus. There is a need to elucidate the possible adjuvant effects of chitosan NPs and the immunological mechanisms of protective immunity induced by OMPs administered orally to fish. PMID:27827990
Dubey, Saurabh; Avadhani, Kiran; Mutalik, Srinivas; Sivadasan, Sangeetha Madambithara; Maiti, Biswajit; Girisha, Shivani Kallappa; Venugopal, Moleyur Nagarajappa; Mutoloki, Stephen; Evensen, Øystein; Karunasagar, Indrani; Munang’andu, Hetron Mweemba
The use of oral vaccination in finfish has lagged behind injectable vaccines for a long time as oral vaccines fall short of injection vaccines in conferring protective immunity. Biodegradable polymeric nanoparticles (NPs) have shown potential to serve as antigen delivery systems for oral vaccines. In this study the recombinant outer membrane protein A (rOmpA) of Edwardsiella tarda was encapsulated in chitosan NPs (NP-rOmpA) and used for oral vaccination of Labeo fimbriatus. The rOmpA purity was 85%, nanodiameter <500 nm, encapsulation efficiency 60.6%, zeta potential +19.05 mV, and there was an in vitro release of 49% of encapsulated antigen within 48 h post incubation in phosphate-buffered saline. Empty NPs and a non-formulated, inactivated whole cell E. tarda (IWC-ET) vaccine were used as controls. Post-vaccination antibody levels were significantly (p = 0.0458) higher in the NP-rOmpA vaccinated fish (Mean OD450 = 2.430) than in fish vaccinated with inactivated whole cell E. tarda (IWC-ET) vaccine (Mean OD450 = 1.735), which corresponded with post-challenge survival proportions (PCSP) of 73.3% and 48.28% for the NP-rOmpA and IWC-ET groups, respectively. Serum samples from NP-rOmpA-vaccinated fish had a higher inhibition rate for E. tarda growth on tryptic soy agar (TSA) than the IWC-ET group. There was no significant difference (p = 0.989) in PCSPs between fish vaccinated with empty NPs and the unvaccinated control fish, while serum from both groups showed no detectable antibodies against E. tarda. Overall, these data show that the NP-rOmpA vaccine produced higher antibody levels and had superior protection over the IWC-ET vaccine, showing that encapsulating OmpA in chitosan NPs confer improved protection against E. tarda mortality in L. fimbriatus. There is a need to elucidate the possible adjuvant effects of chitosan NPs and the immunological mechanisms of protective immunity induced by OMPs administered orally to fish.
Padhi, B K; Ghosh, S K; Mandal, R K
We have cloned and characterized two highly reiterated, tandemly repeated, and A+T rich MboI DNA fragments, one in Cirrhina mrigala (Cyprinidae), with a monomer size of 266 bp, and one in Clarias batrachus (Clariidae), with a monomer size of 227 bp. The MboI fragment in C. mrigala is species-specific and absent in other carps, such as Catla catla and Labeo rohita. The MboI fragment in C. batrachus was also present in two other catfishes tested, namely Clarias gariepinus and Heteropneustes fossilis. In C, mrigala x C. catla and C. mrigala x L. rohita hybrids, the C. mrigala specific MboI fragment is inherited uniparentally. In the reciprocal hybrids of C. batrachus x H. fossilis, the satellite ladder contains the bands of both parental species. The MboI satellite of carp may be useful in genetic introgression analysis and that of catfish in distinguishing between gynogenetic progeny and true hybrids.
Venugopal, T; Mathavan, S; Pandian, T J
A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences of Labeo rohita, Cirrhina mrigala and Catla catla have been cloned, characterized and overexpressed in Escherichia coli. These sequences show 96-98% homology to each other and are about 85% homologous to that of common carp. Besides, an attempt has been made for the first time to describe a 3-D model of the fish GH protein.
Jarapala, Sreenivasa Rao; Kandlakunta, Bhaskarachary; Thingnganing, Longvah
The objective of the present study was to investigate trace metal levels of different varieties of fresh water fish using Inductively Coupled Plasma Mass Spectrophotometer after microwave digestion (MD-ICPMS). Fish samples were collected from the outlets of twin cities of Hyderabad and Secunderabad. The trace metal content in different varieties of analyzed fish were ranged from 0.24 to 1.68 mg/kg for Chromium in Cyprinus carpio and Masto symbollon, 0.20 to 7.52 mg/kg for Manganese in Labeo rohita and Masto symbollon, 0.006 to 0.07 mg/kg for Cobalt in Rastrelliger kanagurta and Pampus argenteus, 0.31 to 2.24 mg/kg for Copper in Labeo rohita and Penaeus monodon, 3.25 to 14.56 mg/kg for Zinc in Cyprinus carpio and Macrobrachium rosenbergii, and 0.01 to 2.05 mg/kg for Selenium in Rastrelliger kanagurta and Pampus argenteus, respectively. Proximate composition data for the different fishes were also tabulated. Since the available data for different trace elements for fish is scanty, here an effort is made to present a precise data for the same as estimated on ICP-MS. Results were in accordance with recommended daily intake allowance by WHO/FAO. PMID:24744789
Jarapala, Sreenivasa Rao; Kandlakunta, Bhaskarachary; Thingnganing, Longvah
The objective of the present study was to investigate trace metal levels of different varieties of fresh water fish using Inductively Coupled Plasma Mass Spectrophotometer after microwave digestion (MD-ICPMS). Fish samples were collected from the outlets of twin cities of Hyderabad and Secunderabad. The trace metal content in different varieties of analyzed fish were ranged from 0.24 to 1.68 mg/kg for Chromium in Cyprinus carpio and Masto symbollon, 0.20 to 7.52 mg/kg for Manganese in Labeo rohita and Masto symbollon, 0.006 to 0.07 mg/kg for Cobalt in Rastrelliger kanagurta and Pampus argenteus, 0.31 to 2.24 mg/kg for Copper in Labeo rohita and Penaeus monodon, 3.25 to 14.56 mg/kg for Zinc in Cyprinus carpio and Macrobrachium rosenbergii, and 0.01 to 2.05 mg/kg for Selenium in Rastrelliger kanagurta and Pampus argenteus, respectively. Proximate composition data for the different fishes were also tabulated. Since the available data for different trace elements for fish is scanty, here an effort is made to present a precise data for the same as estimated on ICP-MS. Results were in accordance with recommended daily intake allowance by WHO/FAO.
Studies on Trichodinid and Chilodonellid Ciliophorans (Protozoa: Ciliophora) in the Indian freshwater and estuarine fishes with description of Trichodinella sunderbanensis sp. nov. and Trichodina nandusi sp. nov.
Mitra, Amlan Kumar; Bandyopadhyay, Probir K; Gong, Yingchun
A comprehensive icthyoparasotological survey among estuarine and freshwater fishes of West Bengal, India revealed Trichodinella sunderbanensis sp. nov. and Trichodina acuta Lom, 1961 from an estuarine fish Mystus gulio (Hamilton, 1822) from the Vidyadhari river; Trichodina nandusi sp. nov. from a freshwater fish, Nandus nandus (Hamilton-Buchanan); Chilodonella hexasticha (Kiernik, 1909) Kahl, 1931 from freshwater fishes Labeo rohita (Hamilton, 1822); and Cyprinus carpio Linnaeus, 1758 from the Rupnarayan river. Taxonomic description of all the species based on wet silver nitrate impregnation method along with additional comments based on scanning electron microscopic descriptions of T. nandusi sp. nov. are also provided in this paper. Prevalence rate, morphometric parameters and comparisons with closely related species are also discussed.
Dar, Shoaib Ali; Kaur, Harpreet; Chishti, M Z
In the study, a new species, Myxobolus chushi n. sp. infecting gills of wild specimens of Schizothorax niger (Heckel) inhabiting Wullar Lake in Kashmir Himalayas, (J&K) India has been described based on morphology of the myxospore and using partial 18S rDNA sequencing. Pathological changes in the gills have been studied with the help of histological sections stained with Luna's method. Twenty fish specimens were examined, out of which four had oval, white plasmodia in gills measuring 2.0×0.5mm. The myxospores were spherical to ovoidal in shape with slightly attenuated posterior end, measuring 11.17±0.23 (10.60-11.40)μm in length and 9.14±0.06 (8.80-9.20)μm in width, having a prominent pore at the anterior end. The polar capsules were pyriform in shape, measuring 4.25±0.15 (4.00-4.40)μm in length and 2.38±0.27 (2.00-2.65)μm in width having polar filaments forming coils up to 5 in number. Parietal folds 9 in number present on the posterior part of the shell. The intensity of infection was recorded to be moderate as indicated by gill plasmodial index (GPI=2). The plasmodium was located in the vascular network occupying whole of the gill lamella therefore typed as intralamellar vascular type, LV3. Analysis of 18S small subunit (SSU) rDNA sequence of the isolate demonstrated 90% homogeneity with M. sp. KLT-2014 infecting scales of Labeo rohita from Myanmar and 89% with M. dermiscalis infecting scales of Labeo rohita from India.
Dar, Shoaib Ali; Kaur, Harpreet; Chishti, M Z
The present study was aimed to isolate myxozoan parasites infecting the native carp Schizothorax esocinus (Heckel) and pond reared carp Labeo rohita (Ham.) collected from fresh water ecosystems of Jammu and Kashmir from July 2013 to June 2014. Various organs such as eyes, gills, scales, fins, intestines, visceral organs and pericardial cavity of as many as 100 live specimens were examined. Results revealed that 14% of the examined L. rohita were infected with Myxobolus rocatlae (Ham.) and 6% of S. esocinus (Heckel) were infected with M. kashmirensis sp. nov. Clinical signs revealed mucous laden gills while the parasitological observation under stereozoom binocular microscope revealed multiple minute plasmodia on the gills. On rupturing these plasmodia with sharp needle 100-500 myxospores were liberated. The identification to species level was done on the basis of morphological and morphometric attributes of the myxospores. This is the first record of myxozoan parasites in fishes from the state of Jammu and Kashmir, India. Histological observations of infected gills revealed plasmodia of two types (LV4 intralamellar vascular type) in M. rocatlae and (LV3 intralamellar vascular type) in M. kasmirensis causing necrosis, hyperplasia, hypertrophy and vacuolization of the epithelial and vascular endothelium.
Mondal, Debjit Kumar; Kaviraj, Anilava
Severe deterioration of water quality occurs during jute retting in ponds, canals, floodplain lakes, and other inland water bodies in the rural areas of West Bengal in India. Attempts were made to evaluate changes in the physicochemical parameters of water caused by jute retting, and their impact on the survival of two species of freshwater fish (Labeo rohita and Hypophthalmicthys molitrix) and two species of freshwater invertebrate (Daphnia magna, a Cladocera, and Branchiura sowerbyi, an Oligochaeta). Results showed that jute retting in a pond for 30 days resulted in a sharp increase in the BOD (>1,000 times) and COD (>25 times) of the water, along with a sharp decrease in dissolved oxygen (DO). Free CO(2), total ammonia, and nitrate nitrogen also increased (three to five times) in water as a result of jute retting. Ninety-six-hour static bioassays performed in the laboratory with different dilutions of jute-retting water (JRW) revealed that D. magna and B. sowerbyi were not susceptible to even the raw JRW whereas fingerlings of both species of fish were highly susceptible, L. rohita being more sensitive (96 h LC(50) 37.55% JRW) than H. molitrix (96 h LC(50) 57.54% JRW). Mortality of fish was significantly correlated with the percentage of JRW.
Ghosh, Subarna; Bandyopadhyay, P K
Three new species of Myxobolus are described from the mucous membrane around gill lamellae of carps of West Bengal. The new species have been named as M. muralidharani sp. n., M. nilimae sp. n. from Labeo rohita, and M. harpreetae sp. n. from Labeo bata respectively. Spores of M. muralidharani sp. n. measured 13.84 ± 0.049 (13-14) × 4.95-6 (5.35) µm, with two elongated equal sized polar capsules. Spores of M. nilimae sp. n. measured 15.9-17.1 (16.65) µm × 6.2-7.5 (6.9) µm, with two equal petal shaped polar capsules, 9.0-11 (10.4) µm × 3-3.5 (3.16) µm. In contrast to aforementioned two new species, M. harpreetae sp. n. had two unequal polar capsules, 8.7-9.2 (9.149) µm × 2.9-4.0 (3.45) µm and 7.6-8.3 (7.94) µm × 2.8-3.3 (2.9) µm in each spore, 11.4-13.8 (13.2) µm × 4.9-7.8 (6.51) µm.
Ghosh, Subarna; Bandyopadhyay, P K
Myxobolus Bütschli, 1882 (Myxozoa, Myxosporea, Bivalvulida, Myxobolidae) is an important parasitic protozoan of freshwater fishes reported from almost all over the world. The severity of infection may lead to mortality of the host fish. The present paper deals with the description of three new species of Myxobolus Bütschli, 1882, Myxobolus sonarpurensis sp. nov., Myxobolus elongatum sp. nov. and Myxobolus petalum sp. nov. from a freshwater minor carp Labeo bata (Hamilton, 1822) and major carp Labeo rohita (Hamilton, 1822) respectively from West Bengal, India. Spores of Myxobolus sonarpurensis sp. nov. measures 12.16 µm × 6.74 µm, with two elongated unequal sized polar capsules measuring, 8.84 µm × 3.019 µm and 7.71 µm × 3.12 µm. Spores of Myxobolus elongatum sp. nov. are 16.23 µm × 7.9 µm with two equal polar capsules 8.896 µm × 3.468 µm and 6.51 µm × 3.46 µm. In comparison to aforementioned two new species, Myxoboluspetalum sp. nov. have two equal polar capsules, 7.905 µm × 2.975 µm (2.9 µm) in each spore, 12.07 µm × 5.95 µm.
Al-Yakoob, S.; Bahloul, M. ); Bou-Olayan, A.H.
Complexation of metals by coordinate linkages with appropriate organic molecules in biological tissues is an important process involved in metal accumulation by aquatic organisms. Fish respiratory systems differ from all other systems because damage to gills has immediate impacts on the rest of the fish's body. Veer et al. observed significant correlation between gill-metal concentration and whole-body weight. More nickel is accumulated in gill tissue of the catfish (Clarias batrachus) than in the liver or intestine. More cadmium is accumulated in gill tissue of the fish Heteropneustes fossilis (Bloch) and Channa punctatus (Bloch) than in the liver or kidney. When exposed to lethal and sublethal concentrations of copper, gills of the freshwater fish Labeo rohita (Hamilton) showed the highest degree of copper accumulation. Petroleum and petrochemical industry wastes contribute significantly to metal enrichment of the Arabian Gulf marine environment. Because accumulation of metal ions is significant in gills, levels of Cd, Cr, Cu, Ni and Pb were investigated in gills of fish from potentially impacted areas along the western side of the Arabian Gulf after the 1991 oil-spill. 15 refs., 3 figs., 1 tab.
Yasmeen, Kousar; Mirza, Muhammad Aslam; Khan, Namra A; Kausar, Nazish; Rehman, Atta-Ur; Hanif, Muddasir
Fish is a vital food for humans and many animals. We report an environmental monitoring study to assess the trace metals in fish species caught from Arabian Sea and commercially available in the coastal city Karachi, Pakistan. Heavy metals such as copper, iron, lead and cadmium were determined in the skin, fillet and heart of the fish species Pampus argenteus, Epinephelus chlorostigma, Rachycentron canadum, Scomberomorus commerson, Johnius belangerii, Labeo rohita, Lutjanus argentimaculatus, Trachinotus blochii, Pomadsys olivaceum and Acanthopagrus berda by the atomic absorption spectrophotometer. The concentration (mg kg(-1), dry weight) range was: Cd (0.00-0.041), Cu (0.006-0.189), Fe (0.413-4.952) and Pb (0.00-0.569). Cadmium, copper and iron levels were below the tolerable limits whereas concentration of lead in the skins of S. commerson, E. chlorostigma, J. belangerii, A. berda; L. argentimaculatus, fillets of J. belangerii, E. chlorostigma and in the heart of J. belangerii exceeded the recommended limits. Therefore fish skin should be discouraged as food for humans or animals. The results indicate that a number of fish species have higher concentration of heavy metals dangerous for human health. Since the fish P. olivaceum (Dhotar) has the lowest level of trace metals therefore we recommend it for breeding and human consumption.
Panda, Somerita; Ghosh, Subarna; Bandyopadhyay, Probir Kumar
In India, more than 104 species of Myxobolus have so far been reported infecting freshwater as well as marine fishes. The study focuses on the description of a new myxosporean species, Myxobolus sp.n PKB 2014 from the gill lamellae of an Indian major carp Labeo rohita. The species have been described on the basis of morphological characterization of the spores, tissue architecture and 18S rDNA sequence data. The plasmodia of Myxobolus sp.n PKB 2014 were round in shape measuring, 50 to 70 μm in diameter and spores were ellipsoidal in frontal view measures 14.7 ± 0.51 μm. The 18S rRNA nucleotide sequence with 806 bp of Myxobolus sp.n PKB 2014 (Accession number KJ652226) clustered phylogenitically with other Myxobolus spp. infecting cyprinid gills with 90-99% similarity. According to the phylogenetic study we concluded that M. wulli was the closest relative having 99% similarity with the species under description but the sequence was distinct in each species which additionally exhibited different morphological features. The infection rate was low to moderate. After through comparison it can be concluded that the species being described here is new to science which is designated as Myxobolus sp.n. PKB 2014. PMID:27844012
Sinha, Tanur; Ahmaruzzaman, M
The present work reports the utilization of a common household waste material (fish scales of Labeo rohita) for the synthesis of copper nanoparticles. The method so developed was found to be green, environment-friendly, and economic. The fish scale extracts were acting as a stabilizing and reducing agents. This method avoids the use of external reducing and stabilizing agents, templates, and solvents. The compositional abundance of gelatin may be envisaged for the effective reductive as well as stabilizing potency. The mechanisms for the formation of nanoparticles have also been presented. The synthesized copper nanoparticles formed were predominantly spherical in nature with an average size of nanoparticles in the range of 25-37 nm. The copper nanoparticles showed characteristic Bragg's reflection planes of fcc which was supported by both selected area electron diffraction and X-ray diffraction pattern and showed surface plasmon resonance at 580 nm. Moreover, the energy dispersive spectroscopy pattern also revealed the presence of only elemental copper in the copper nanoparticles. The prepared nanoparticles were used for the remediation of a carcinogenic and noxious textile dye, Methylene blue, from aqueous solution. Approximately, 96 % degradation of Methylene blue dye was observed within 135 min using copper nanoparticles. The probable mechanism for the degradation of the dye has been presented, and the degraded intermediates have been identified using the liquid chromatography-mass spectroscopy technique. The high efficiency of nanoparticles as photocatalysts has opened a promising application for the removal of hazardous dye from industrial effluents contributing indirectly to environmental cleanup process.
Adhikari, Subhendu; Sahu, Bharat Chandra; Dey, Lambodar
Experiments were conducted for the study of nutrient budget in farmers' ponds (0.4-0.6 ha) in Orissa, India, at stocking densities of 0.30-0.38/m(2) for scampi (Macrobrachium rosenbergii) and 0.60-0.70/m(2) for Indian major carps (Catla catla, Labeo rohita and Cirrhinus mrigala). The average initial body weights of scampi and the major carps were 0.02 and 8-10 g, respectively. The culture period was 272-292 days. Feed was the main input. The FCR (feed conversion ratio) varied from 1.78 to 1.83. Feed and cow dung were applied to these ponds as organic inputs. At harvest time, the average weight of scampi and carps varied from 73 to 92 g and from 718 to 820 g, respectively. Among all the inputs, feed alone accounted for 97.60% total nitrogen (N), 97.90% total phosphorus (P) and 94.72% total organic carbon (OC), respectively. The harvest of scampi and carps accounted for recovery of 52.45% N, 19.43% P and 18.12% OC, respectively. N, P and OC accumulated in sediment were 38.31, 71.40 and 69.50%, respectively. The median nutrient loads in the fish pond effluents were equivalent to 0.58 kg of inorganic N, 0.135 kg of P and 8.83 kg of total OC per ton of Indian major carps and scampi production.
Tiwari, Sudhanshu; Singh, Ajay
Active compound oleandrin extracted from Nerium indicum (Lal Kaner) leaf has potent piscicidal activity. The piscicidal activity of oleandrin on freshwater fish C. punctatus was both time and dose dependent. Exposure to sub-lethal doses of oleandrin for 24hr and 96hr to fish caused significant alteration in the level of total protein, total free amino acid, nucleic acid, glycogen, pyruvate, lactate and enzyme protease, phosphatases, alanine aminotransferase, aspartate aminotransferase and acetylcholinesterase activity in liver and muscle tissues. The alterations in all the above biochemical parameters were also significantly time and dose dependent. The results show a significant recovery in all the above biochemical parameters, in both liver and muscle tissues of fish after the 7th day of the withdrawal of treatment. Toxicity persistence test of oleandrin on juvenile Labeo rohita shows that fish seed of common culturing carp can be released into rearing ponds after three days of oleandrin treatment. It supports the view that the oleandrin is safer and may be useful substitute of other piscicides for removing the unwanted freshwater fishes from aquaculture ponds.
Sood, Neeraj; Chaudhary, Dharmendra K; Rathore, Gaurav; Singh, Akhilesh; Lakra, W S
Snakehead Channa striata is an important freshwater food fish in many Southeast Asian countries. Three monoclonal antibodies (C9, C10 and D10) were developed against purified serum immunoglobulins of Channa striata (Cs-Ig) and characterized. C9 and D10 MAbs were specific to heavy chain, while C10 MAb detected only unreduced Cs-Ig in western blotting. In competitive ELISA, C9 and C10 MAbs were specific to C. striata Ig and showed no cross reactivity with serum Ig of other fish species i.e. Channa punctatus, Channa marulius, Clarias batrachus and Labeo rohita. D10 MAb showed reactivity to serum Ig of C. striata and C. marulius. In FACS analysis of gated lymphocytes, the percentage of Ig+ cells detected by C9 MAb was 18.2%, 27.7% and 10.3% in blood, spleen and kidney, respectively (n=3, body weight 500-600 g). However, only a few cells (0.5%) were found to be Ig+ in thymus (n=5). C9 MAb was also successfully employed to demonstrate Ig+ cells in blood smears and formalin fixed sections of spleen and kidney. These findings suggest that the spleen plays an important role in humoral immunity as compared to head kidney. Further, these MAbs can be useful immunological tool in monitoring health status of cultured C. striata.
Banerjee, Goutam; Nandi, Ankita; Ray, Arun Kumar
In the present investigation, probiotic potential (antagonistic activity, enzyme production, hemolytic activity, biosafety, antibiotic sensitivity and bile tolerance level) of Bacillus subtilis LR1 was evaluated. Bacteriocin produced by the bacterial strain B. subtilis LR1 isolated from the gastrointestinal tract of Labeo rohita was purified and characterized. The molecular weight of the purified bacteriocin was ~50 kDa in 12 % Native PAGE and showed inhibitory activity against four fish pathogens such as Bacillus mycoides, Aeromonas salmonicida, Pseudomonas fluorescens and Aeromonas hydrophila. The purified bacteriocin was maximally active at temperature 40 °C and pH 7.0, while none of the tested surfactants affect the bacteriocin activity. Extracellular enzyme activity of the selected bacterial strain was also evaluated. Amylase activity was estimated to be highest (38.23 ± 1.15 µg of maltose liberated mg(-1) protein ml(-1) of culture filtrate) followed by cellulase and protease activity. The selected bacterium was sensitive to most of the antibiotics used in this experiment, can tolerate 0.25 % bile salt and non-hemolytic in nature. Finally, the efficiency of the proposed probiotic candidate was evaluated in in vivo condition. It was detected that the bacterial strain can effectively reduce bacterial pathogenicity in Indian major carps.
Adhikari, S; Ghosh, L; Rai, S P; Ayyappan, S
The concentrations of lead, cadmium, chromium, copper, and zinc were investigated in the sewage-fed pond water, sediment, and the various organs of Labeo rohita, Catla catla, Cirrhinus mrigala, Oreochromis mossambicus, and Cyprinus carpio cultured in sewage-fed ponds, Kolkata, India. Among the metals, cadmium, lead, and zinc were detected in water and, except lead, were below the water quality guideline levels for the protection of freshwater aquatic life proposed by CEQG (Canadian Environmental Quality Guidelines) and AENV (Alberta Environment). Therefore, lead could pose danger to aquatic organisms. All the five metals were detected in the sediment and, except cadmium and lead, were below the sediment quality guideline levels for aquatic life proposed by EPA (Environmental Protection Agency). Therefore, these two metals could be toxic to aquatic organisms. Significant (P > 0.05) differences were observed among the five fish species for all these metals accumulation. Also, significant (P > 0.05) differences were noticed among these metals accumulation in fish organs. Cadmium showed the least bioaccumulation, while zinc showed the highest bioaccumulation in all the fish species. Though the metal concentration in the different fish tissues was variable, the highest concentration was found in kidney and the lowest in the muscle. Concentrations of these metals in the muscle tissue of all the fish species were well below the consumption safety tolerance in fish set by WHO/FAO, and thus, so far as these metals are concerned, these sewage-fed cultured fishes are safe and suitable for human consumption.
Gupta, Bina; Kumar, Rahul; Rani, Manviri; Agarwal, Tripti
This paper deals with the dynamics of chromium, nickel, copper and lead among the different components namely water, surface sediments, submerged and free floating macrophytes and fish of the twin manmade lakes, Upper and Lower lakes, of Bhopal (M.P., India). Some basic parameters of water and sediment have also been studied. The basin of the lake system is densely populated and the water is used for various purposes including drinking. Ni and Pb along with nitrate in both lakes are significantly higher than the drinking water quality criteria of USEPA. The concentration of the metals in the sediments is noticeably higher than that present in the adjoining rock, particularly Ni and Pb. There is a significant uptake of metals by the macrophytes (Eichhornia crassipes and Hydrilla verticillata) and fish (Labeo rohita and Oreochromis niloticus) mainly in summer. The fish of Lower lake (O. niloticus) is unfit for human consumption. The data have been statistically treated. Principle component analysis and cluster analysis were performed to define the origin of metals and to assess the relationship among the sites. Overall the Lower lake is more polluted than the Upper lake. In aggregate, the lake system is under an environmental stress due to certain practices.
Verma, Dev Kumar; Rathore, Gaurav
Abstract Yellow pigmented, filamentous, Gram-negative bacteria belonging to genus Flavobacterium are commonly associated with infections in stressed fish. In this study, inter-species diversity of Flavobacterium was studied in apparently healthy freshwater farmed fishes. For this, ninety one yellow pigmented bacteria were isolated from skin and gill samples (n = 38) of three farmed fish species i.e. Labeo rohita, Catla catla and Cyprinus carpio. Among them, only twelve bacterial isolates (13.18%) were identified as Flavobacterium spp. on the basis of morphological, biochemical tests, partial 16S rDNA gene sequencing and phylogenetic analysis. On the basis of 16S rDNA gene sequencing, all the 12 isolates were 97.6-100% similar to six different formally described species of genus Flavobacterium. The 16S rDNA based phylogenetic analysis grouped these strains into six different clades. Of the 12 isolates, six strains (Fl9S1-6) grouped with F. suncheonense, two strains (Fl6I2, Fl6I3) with F. indicum and the rest four strains (Fl1A1, Fl2G1, Fl3H1 and Fl10T1) clustered with F. aquaticum, F. granuli, F. hercynium and F. terrae, respectively. None of these species except, F. hercynium were previously reported from fish. All the isolated Flavobacterium species possessed the ability of adhesion and biofilm formation to colonize the external surface of healthy fish. The present study is the first record of tropical freshwater farmed fishes as hosts to five environmentally associated species of the Flavobacterium. PMID:26691454
Farah-Ayuni, F.; Muse, A. O.; Samat, A.; Shukor, M. N.
Hard plates covering fish body or scales, do not only serve as external protection, but are also important as one of the tools for species identification. This study was designed to recognise morphological characters of fish scales that can be used for identification and to comprehend the inter-specific variations of scales from different species by using selected indices from scale measurements; L1/1, L2/L, L1/L2, and W/L. Sixteen species of common freshwater fishes were sampled from a few localities in Peninsular Malaysia. Examinations were based on photographs to determine the scale morphologies. Basic distinguishable characteristics that were identified are type of scale, the overall shape, the position and shape of the focus, distribution of radii, and the circuli appearance. It was found that scale type and radii arrangement at the anterior field are two significant morphological features that could distinguish between Cyprinidae and other families. Main characters used to identify scales at species level are radii cover, structure of inter radial tongue, and radii arrangement. Meanwhile, multiple comparison tests to reveal inter-specific variations suggested that there are significant differences among 16 species of common freshwater fishes studied. Two indices were found to be suitable to highlight the significant differences (P<0.05) for all species which are index L1/L2 and index W/L. For all indices, species that showed significantly different with most of other species are Osteochilus vittatus, Pristolepis fasciata, Rasbora sumatrana, and Labeo rohita. The scales of Osteochilus vittatus showed significant values with most of other species in more than one index; index L1/L and index L2/L. The distinctive morphological character that distinguished it from other Cyprinidae species is having clearly formed serrations in the middle of the inter-radial tongue at the anterior region.
Singh, Arvind R.; Ferosekhan, S.; Kothari, Dushyant C.; Pal, Asim Kumar; Jadhao, Sanjay Balkrishna
Exogenous proteolytic enzyme supplementation is required in certain disease conditions in humans and animals and due to compelling reasons on use of more plant protein ingredients and profitability in animal feed industry. However, limitations on their utility in diet are imposed by their pH specificity, thermolabile nature, inhibition due to a variety of factors and the possibility of intestinal damage. For enhancing the efficacy and safety of exogenous trypsin, an efficient chitosan (0.04%) nanoencapsulation-based controlled delivery system was developed. An experiment was conducted for 45 days to evaluate nanoencapsulated trypsin (0.01% and 0.02%) along with 0.02% bare trypsin and 0.4% chitosan nanoparticles against a control diet on productive efficiency (growth rate, feed conversion and protein efficiency ratio), organo-somatic indices, nutrient digestibility, tissue enzyme activities, hematic parameters and intestinal histology of the fish Labeo rohita. All the synthesized nanoparticles were of desired characteristics. Enhanced fish productive efficiency using nanoencapsulated trypsin over its bare form was noticed, which corresponded with enhanced (P<0.01) nutrient digestibility, activity of intestinal protease, liver and muscle tissue transaminases (alanine and aspartate) and dehydrogenases (lactate and malate), serum blood urea nitrogen and serum protein profile. Intestinal tissues of fish fed with 0.02% bare trypsin showed broadened, marked foamy cells with lipid vacuoles. However, villi were healthier in appearance with improved morphological features in fish fed with nanoencapsulated trypsin than with bare trypsin, and the villi were longer in fish fed with 0.01% nanoencapsulated trypsin than with 0.02% nanoencapsulated trypsin. The result of this premier experiment shows that nanoencapsulated trypsin mimics zymogen-like proteolytic activity via controlled release, and hence the use of 0.01% nanoencapsulated trypsin (in chitosan nanoparticles) over bare
Verma, Dev Kumar; Rathore, Gaurav
Yellow pigmented, filamentous, Gram-negative bacteria belonging to genus Flavobacterium are commonly associated with infections in stressed fish. In this study, inter-species diversity of Flavobacterium was studied in apparently healthy freshwater farmed fishes. For this, ninety one yellow pigmented bacteria were isolated from skin and gill samples (n = 38) of three farmed fish species i.e. Labeo rohita, Catla catla and Cyprinus carpio. Among them, only twelve bacterial isolates (13.18%) were identified as Flavobacterium spp. on the basis of morphological, biochemical tests, partial 16S rDNA gene sequencing and phylogenetic analysis. On the basis of 16S rDNA gene sequencing, all the 12 isolates were 97.6-100% similar to six different formally described species of genus Flavobacterium. The 16S rDNA based phylogenetic analysis grouped these strains into six different clades. Of the 12 isolates, six strains (Fl9S1-6) grouped with F. suncheonense, two strains (Fl6I2, Fl6I3) with F. indicum and the rest four strains (Fl1A1, Fl2G1, Fl3H1 and Fl10T1) clustered with F. aquaticum, F. granuli, F. hercynium and F. terrae, respectively. None of these species except, F. hercynium were previously reported from fish. All the isolated Flavobacterium species possessed the ability of adhesion and biofilm formation to colonize the external surface of healthy fish. The present study is the first record of tropical freshwater farmed fishes as hosts to five environmentally associated species of the Flavobacterium.
Sinha, Tanur; Ahmaruzzaman, M
Green and facile synthetic methods have gained marvellous fame for the production of polyhedral, anisotropic and spherical gold, and gold-silver bimetallic nanostructures. The useful pivotal characteristics of a green procedure are the usage of environment benign solvent medium, reducing and stabilising agents, and shorter reaction time. We describe here a novel, and greener method for the production of gold and gold-silver core shell nanostructures using aqueous fish scales extract of the Labeo rohita. The effect of various reaction parameters, such as temperature and concentration for the synthesis of the nanostructures were studied. Results indicated that triangular and decahedron gold nanostructures were formed at a lower temperature (40°C) and concentration (10%). While, icosahedral and spherical gold nanostructures were produced at a comparatively higher temperature (100°C) and concentration (40%). The study also revealed that the core-shell bimetallic nanostructures with different morphologies (spherical and oval-shape) were formed at different ratios of chloroaurate and silver nitrate solution. Thus, the present study indicated a simple shape controlled synthesis of gold and gold silver core-shell nanostructures. The synthesised gold nanotriangles were coated over the glass substrate and found to be highly efficient in absorbing infra-red radiations for potential architectural applications. Therefore, the study demonstrated the facile usage of gold nanotriangles for optical coatings. The present strategy depicted the dual functional ability of the fish scale extract as reducing and stabilising agents. This strategy also eliminates the usage of hazardous chemicals, toxic solvents and harsh reducing and stabilizing agents.
Sood, Neeraj; Chaudhary, Dharmendra K; Singh, Akhilesh; Rathore, Gaurav
Serum immunoglobulins of Clarias batrachus (Cb-Ig) were purified by affinity chromatography using bovine serum albumin as capture ligand. Under reducing conditions in SDS-PAGE, Cb-Ig was composed of a heavy (H) chain (68.7 kDa) and two light (L) chains (27.4 and 26.3 kDa). Purified Cb-Ig was used to produce a monoclonal antibody (MAb) designated E4 MAb that belonged to IgG1 subclass. In Western blotting, this MAb showed binding to H chain of purified Cb-Ig and putative H chains in reduced sera of C. batrachus, Clarias gariepinus and Heteropneustes fossilis. However, no binding was observed with serum protein of Labeo rohita and Channa striata. Cross-reactivity of anti-Cb-Ig MAb was observed with serum of C. batrachus, C. gariepinus and H. fossilis in competitive ELISA. In immunoblotting of non-reduced Cb-Ig with E4 MAb, four bands assumed to be tetrameric, trimeric, dimeric and monomeric form were observed. In flow cytometric analysis of the gated lymphocytes, the number of surface Ig-positive (Ig+) cells in blood, spleen, kidney and thymus of C. batrachus was determined to be 50.1 ± 3.1, 55.1 ± 3.36, 42.4 ± 4.81 and 5.1 ± 0.89%, respectively, using E4 MAb. Ig+ cells were also demonstrated in formalin-fixed paraffin embedded tissue sections of spleen, kidney, thymus and smears of blood mononuclear cells in indirect immunoperoxidase test. The developed MAb was employed to detect pathogen-specific immunoglobulins in the sera of C. batrachus immunized with killed Edwardsiella tarda, by an indirect ELISA. This monoclonal antibody can be useful tool in immunological research and assays.
Masih, Prachi; Luhariya, Rupesh K; Das, Rakhi; Gupta, Arti; Mohindra, Vindhya; Singh, Rajeev K; Srivastava, Rohit; Chauhan, U K; Jena, J K; Lal, Kuldeep K
This study is aimed to identify polymorphic microsatellite markers and establish their potential for population genetics studies in three carp (family cyprinidae; subfamily cyprininae) species, Labeo rohita, Catla catla and Cirrhinus mrigala through use of cyprinid primers. These species have high commercial value and knowledge of genetic variation is important for management of farmed and wild populations. We tested 108 microsatellite primers from 11 species belonging to three different cyprinid subfamilies, Cyprininae, Barbinae and Leuciscinae out of which 63 primers (58.33%) successfully amplified orthologous loci in three focal species. Forty-two loci generated from 29 primers were polymorphic in these three carp species. Sequencing of amplified product confirmed the presence of SSRs in these 42 loci and orthologous nature of the loci. To validate potential of these 42 polymorphic loci in determining the genetic variation, we analyzed 486 samples of three focal species collected from Indus, Ganges and Brahmaputra river systems. Results indicated significant genetic variation, with mean number of alleles per locus ranging from 6.80 to 14.40 and observed heterozygosity ranging from 0.50 to 0.74 in the three focal species. Highly significant (P < 0.00001) allelic homogeneity values revealed that the identified loci can be efficiently used in population genetics analysis of these carp species. Further, thirty-two loci from 19 primers were useful for genotyping in more than one species. The data from the present study was compiled with cross-species amplification data from previous results on eight species of subfamily cyprininae to compare cross-transferability of microsatellite loci. It was revealed that out of 226 heterologous loci amplified, 152 loci that originated from 77 loci exhibited polymorphism and 45 primers were of multispecies utility, common for 2-7 species.
Sinha, Tanur; Ahmaruzzaman, M.
Green and facile synthetic methods have gained marvellous fame for the production of polyhedral, anisotropic and spherical gold, and gold-silver bimetallic nanostructures. The useful pivotal characteristics of a green procedure are the usage of environment benign solvent medium, reducing and stabilising agents, and shorter reaction time. We describe here a novel, and greener method for the production of gold and gold-silver core shell nanostructures using aqueous fish scales extract of the Labeo rohita. The effect of various reaction parameters, such as temperature and concentration for the synthesis of the nanostructures were studied. Results indicated that triangular and decahedron gold nanostructures were formed at a lower temperature (40 °C) and concentration (10%). While, icosahedral and spherical gold nanostructures were produced at a comparatively higher temperature (100 °C) and concentration (40%). The study also revealed that the core-shell bimetallic nanostructures with different morphologies (spherical and oval-shape) were formed at different ratios of chloroaurate and silver nitrate solution. Thus, the present study indicated a simple shape controlled synthesis of gold and gold silver core-shell nanostructures. The synthesised gold nanotriangles were coated over the glass substrate and found to be highly efficient in absorbing infra-red radiations for potential architectural applications. Therefore, the study demonstrated the facile usage of gold nanotriangles for optical coatings. The present strategy depicted the dual functional ability of the fish scale extract as reducing and stabilising agents. This strategy also eliminates the usage of hazardous chemicals, toxic solvents and harsh reducing and stabilizing agents.
Background The risks of fish-borne zoonotic trematodes (FZT) to human health constitute an important problem in Vietnam. The infection of humans with these trematodes, such as small liver trematodes (Clonorchis sinensis and Opisthorchis viverrini), intestinal trematodes (Heterophyidae) and others is often thought to be linked to fish culture in areas where the habit of eating raw fish is common. Juvenile fish produced in nurseries are often heavily infected with FZT and since fishes are sold to aquaculture facilities for growth, control of FZT in these fishes should be given priority. Controlling the first intermediate host (i.e., freshwater gastropods), would be an attractive approach, if feasible. The black carp, Mylopharyngodon piceus, is a well-known predator of freshwater snails and is already used successfully for biological control of snails in various parts of the world including Vietnam. Here we report the first trials using it for biological control of intermediate host snails in nursery ponds stocked with 1-week old fry (10–12 mm in length) of Indian carp, Labeo rohita. Methods Semi-field and field experiments were set up to test the effect of black carp on snail populations. In the semi-field experiment a known quantity of snails was initially introduced into a pond which was subsequently stocked with black carp. In the field trial in nursery ponds, density of snails was estimated prior to a nursing cycle and at the end of the cycle (after 9 weeks). Results The results showed that black carp affect the density of snail populations in both semi-field and field conditions. The standing crop of snails in nursery ponds, however, was too high for 2 specimens to greatly reduce snail density within the relatively short nursing cycle. Conclusions We conclude that the black carp can be used in nursery ponds in Northern Vietnam for snail control. Juvenile black carp weighing 100 - 200g should be used because this size primarily prey on intermediate hosts of FZT
Objective To determine the proportion of imported frozen fish contaminated with Salmonella among retail food stores and supermarkets in the Eastern Province of Saudi Arabia. Methods A total of 223 frozen freshwater fish purchased from different supermarkets and grocery stores were analyzed for the presence of foodborne pathogen Salmonella. The isolation of Salmonella was determined and confirmed by using the methods of US Food and Drug Administration's Bacteriological Analytical Manual, CHROMagar Salmonella plus, biochemical tests and API 20E strips. Antimicrobial susceptibilities of Salmonella isolates were determined by the disk diffusion method on Muller-Hinton agar, as described by Kirby-Bauer, in accordance with the guidelines of the Clinical and Laboratory Standards Institute. Results Out of the total 223 fish samples (20 of catfish, 18 of carfu, 20 of mirgal, 25 of milkfish, 35 of mackerel, 75 of tilapia, and 30 of rohu), 89 (39.9%) were tested positive for Salmonella. The prevalence of positive samples were reported for the freshwater fish of pangas (60.0%, n=12), carfu (27.7%, n=5), mirgal (35.0%, n=7), milkfish (52.0%, n=13), mackerel (31.4 %, n=11), tilapia imported from Thailand (64.0%, n=16), tilapia imported from India (28.0%, n=14), rohu imported from Thailand (26.6%, n=4) and rohu imported from Myanmar (46.6%, n=7). A total of 140 isolates of Salmonella spp. were yielded from at least seven different types of frozen freshwater fish imported from 5 different countries and were tested for their susceptibility to 16 selected antimicrobial agents. The highest antibiotic resistance was observed to tetracycline (90.71%) followed by ampicillin (70%) and amoxicillin-clavulanic acid (45%). Conclusions The obtained results of this study shows that these raw retail imported frozen freshwater fish are contaminated with potentially pathogenic Salmonella spp. And the study recommend and suggest that there is a need for adequate consumer measures. PMID:25182443
Gornung, Ekaterina; Colangelo, Paolo; Annesi, Flavia
This paper describes a study of the 5S ribosomal RNA genes (5S rDNA) in a group of 6 species belonging to 4 genera of Mugilidae. In these 6 species, the relatively short 5S rDNA repeat units, generated by PCR and ranging in size from 219 to 257 bp, show a high level of intragenomic homogeneity of both coding and spacer regions (NTS-I). Phylogenetic reconstructions based on this data set highlight the greater phylogenetic and genetic diversity of Mugil cephalus and Oedalechilus labeo compared with the genera Liza and Chelon. Comparative sequence analysis revealed significant conservation of the short 5S rDNA repeat units across Chelon and Liza. Moreover, a second size class of 5S rDNA repeat units, ranging from roughly 800 to 1100 bp, was produced in the Liza and Chelon samples. Only short 5S rDNA repeat units were found in M. cephalus and O. labeo. The sequences of the long 5S rDNA repeat units, obtained in Chelon labrosus and Liza ramada, differ owing to the presence of 2 large insertion/deletions (indels) in the spacers (NTS-II) and show considerable sequence identity with NTS-I spacers. Interspecific sequence variation of NTS-II spacers, excluding the indels, is low. Southern-blot hybridization patterns suggest an intermixed arrangement of short and long repeat units within a single chromosome locus.
F Telli Karakoc A Hewer D H Phillips A F Gaines And G Yuregir
The activities of enzymes associated with xenobiotic metabolism and or oxidative processes, and the levels of aromatic DNA adducts, have been determined in the livers of grey mullet Oedalechilus labeo and Lisa ramada living in two eastern Mediterranean harbours. Glutathione peroxidase GSH P activity was 2.5 times higher 9 IU g-1 liver and glutathione reductase GSSG R activity was twice as high 2.5 IU g-1 liver in fish from the more polluted harbour at Mersin than in the harbour near Erdemli. Superoxide dismutase SOD activity was 25 lower 4.3 IU g-1 liver in the more polluted harbour. The concentrations of glutathione and malondialdehyde varied both with species and environment by a factor of 2.5-3. DNA adducts in liver were determined by 32P postlabelling. In Oedalechilus labeo in the more polluted harbour, adduct levels were 258 21 adducts per 108 nucleotides mean SE; two groups of Lisa ramada were distinguished having 261 48 and 30 6 adducts per 108 nucleotides, respectively. The average adduct level in a group of mullet of mixed species in the less polluted harbour was 3.3 2.3 adducts per 108 nucleotides. The results illuminate the ability of mullet to live in contaminated marine environments, and show that enzyme activities and liver DNA adduct levels can serve as indicators of marine pollution.
Bhattacharyya, Subarna; Chaudhuri, Punarbasu; Dutta, Siddartha; Santra, Subhas Chandra
Total mercury levels were quantified in Tilapia mossambicus, Cirrhinus mrigela and Labio rohita, captured from East Calcutta Wetlands and Titagarh sewage fed aquaculture ponds. The bioconcentration factor of collected fish was assessed. Total mercury level ranged from 0.073 to 0.94 microg/g in both pre and post monsoon season. T. mossambicus in both season and C. mrigela at pre monsoon, cross the Indian recommended maximum limit (0.50 microg/g wet weight) for food consumption and according to World Health Organization guidelines all fish were not recommended for pregnant women and individuals under 15 years ages. A significant correlation was observed between mercury content of aquaculture pond water and fish muscle tissue. Total mercury concentration in experimental sites were higher than the control area (Wilcoxon Ranked-Sum test p > 0.05), which suggested the connection between mercury bioaccumulation and sewage fed aquaculture.
Rossi, Anna Rita; Ungaro, Andrea; De Innocentiis, Sabina; Crosetti, Donatella; Sola, Luciana
The family Mugilidae (Pisces, Mugiliformes) includes species which are present in all tropical and temperate regions. Six species, Chelon labrosus, Mugil cephalus, Liza aurata, L. ramada, L. saliens, Oedalechilus labeo, are commonly found in the Mediterranean. These species have been widely studied through morphological, biochemical, and molecular markers. However, their phylogenetic relationships, and therefore the assumed monophyly of Liza species, still remain unclear: To further investigate this topic, gene-enzyme systems and sequences of the partial 16S rRNA mitochondrial gene were analyzed in Italian samples of all six Mediterranean species. The phylogenetic reconstructions indicated M. cephalus as being the most divergent species and the existence of a main cluster including all the Mediterranean species of Liza and C. labrosus. The parametric bootstrap approach adopted to test alternative phylogenetic hypotheses indicated that the Mediterranean species of Liza do not form a monophyletic group exclusive of Chelon.
Callicó Fortunato, Roberta; Benedito Durà, Vicent; Volpedo, Alejandra
In the Northeastern Atlantic and Mediterranean Sea there are 8 species of the Mugilidae family: Mugil cephalus, Liza aurata, Liza ramada, Oedalechilus labeo, Chelon labrosus, Liza saliens, Liza carinata and Liza haematocheila. The identification of mugilids is very important for local fisheries management and regulations, but it is difficult using gross morphological characters. This work aims to contribute to the identification of mullets present in the Northeastern Atlantic Ocean and Mediterranean Sea using saccular otolith features of each species. Specimens of C. labrosus, L. aurata, L. ramada, L. saliens and M. cephalus were obtained from Delta del Ebro (40°38'N-0°44'E) in artisanal catches. For L. carinata and O. labeo photographs extracted from AFORO online database were used. L. haematocheila was not studied for lack of otolith samples. A general pattern of the saccular otoliths for this family was identified: the shape of the otoliths are rectangular to oblong with irregular margins; they present a heterosulcoid, ostial sulcus acusticus, with an open funnel-like ostium to the anterior margin and a closed, tubular cauda, ending towards the posterior ventral corner, always larger than the ostium. In the present study, the mugilid species could be recognized using their saccular otolith morphology. Here we give the first key to identify Northeastern Atlantic and Mediterranean mullets. The distinctive features between the species were the position and centrality of the sulcus, the curvature of the cauda, the presence of areal depositions and plateaus, and the type of anterior and posterior regions. These features could be used not only to reinforce the identification keys through morphological and meristic characters of the species, but also to identify the species consumed by piscivores, being the otoliths the only identifiable remains of the individuals.
Alam, M. M.; Khan, M. A.; Hussain, M. A.; Moumita, D.; Mazlan, A. G.; Simon, K. D.
Silver carp, Hypopthalmichthys molitrix is one of the most economically valuable fish species in Bangladesh. However, its production is often hindered by parasite-induced mortality. The present study reports the intensity of parasitic infestation in 216 specimens of H. molitrix collected from different fish markets in Rajshahi City, Bangladesh. Nine different parasite species (Trichodina pediculatus, Dactylogyrus vastator, Ichthyophthirius multifilis, Gyrodactylus elegans, Lernaea sp., Apiosoma sp., Myxobolus rohitae, Camallanus ophiocephali, and Pallisentis ophiocephali) were recovered from the gill, skin, stomach, and intestine of host fish. The highest level of infection was observed for host skin, while lower levels were observed for host gill, stomach, and intestine. The results also revealed that the intensity of parasite infection in different organs of H. molitrix varied with the season. In particular, the highest levels of infection were recorded during the winter period (November–February), when fish are most susceptible to parasites. The findings of the study will help in the management and conservation of H. molitrix. PMID:23225858
Alam, M M; Khan, M A; Hussain, M A; Moumita, D; Mazlan, A G; Simon, K D
Silver carp, Hypopthalmichthys molitrix is one of the most economically valuable fish species in Bangladesh. However, its production is often hindered by parasite-induced mortality. The present study reports the intensity of parasitic infestation in 216 specimens of H. molitrix collected from different fish markets in Rajshahi City, Bangladesh. Nine different parasite species (Trichodina pediculatus, Dactylogyrus vastator, Ichthyophthirius multifilis, Gyrodactylus elegans, Lernaea sp., Apiosoma sp., Myxobolus rohitae, Camallanus ophiocephali, and Pallisentis ophiocephali) were recovered from the gill, skin, stomach, and intestine of host fish. The highest level of infection was observed for host skin, while lower levels were observed for host gill, stomach, and intestine. The results also revealed that the intensity of parasite infection in different organs of H. molitrix varied with the season. In particular, the highest levels of infection were recorded during the winter period (November-February), when fish are most susceptible to parasites. The findings of the study will help in the management and conservation of H. molitrix.
Campbell, L M; Osano, O; Hecky, R E; Dixon, D G
Total mercury (THg) concentrations were measured for various fish species from Lakes Turkana, Naivasha and Baringo in the rift valley of Kenya. The highest THg concentration (636 ng g(-1) wet weight) was measured for a piscivorous tigerfish Hydrocynus forskahlii from Lake Turkana. THg concentrations for the Perciformes species, the Nile perch Lates niloticus from Lake Turkana and the largemouth bass Micropterus salmoides from Lake Naivasha ranged between 4 and 95 ng g(-1). The tilapiine species in all lakes, including the Nile tilapia Oreochromis niloticus, had consistently low THg concentrations ranging between 2 and 25 ng g(-1). In Lake Naivasha, the crayfish species, Procambrus clarkii, had THg concentrations similar to those for the tilapiine species from the same lake, which is consistent with their shared detritivore diet. THg concentrations in all fish species were usually consistent with their known trophic position, with highest concentrations in piscivores and declining in omnivores, insectivores and detritivores. One exception is the detritivore Labeo cylindricus from Lake Baringo, which had surprisingly elevated THg concentrations (mean=75 ng g(-1)), which was similar to those for the top trophic species (Clarias and Protopterus) in the same lake. Except for two Hydrocynus forskahlii individuals from Lake Turkana, which had THg concentrations near or above the international marketing limit of 500 ng g(-1), THg concentrations in the fish were generally below those of World Health Organization's recommended limit of 200 ng g(-1) for at-risk groups.
Ahmad, Habib; Yousafzai, Ali Muhammad; Siraj, Muhammad; Ahmad, Rashid; Ahmad, Israr; Nadeem, Muhammad Shahid; Ahmad, Waqar; Akbar, Nazia; Muhammad, Khushi
The contamination of aquatic systems with heavy metals is affecting the fish population and hence results in a decline of productivity rate. River Kabul is a transcountry river originating at Paghman province in Afghanistan and inters in Khyber Pakhtunkhwa province of Pakistan and it is the major source of irrigation and more than 54 fish species have been reported in the river. Present study aimed at the estimation of heavy metals load in the fish living in River Kabul. Heavy metals including chromium, nickel, copper, zinc, cadmium, and lead were determined through atomic absorption spectrophotometer after tissue digestion by adopting standard procedures. Concentrations of these metals were recorded in muscles and liver of five native fish species, namely, Wallago attu, Aorichthys seenghala, Cyprinus carpio, Labeo dyocheilus, and Ompok bimaculatus. The concentrations of chromium, nickel, copper, zinc, and lead were higher in both of the tissues, whereas the concentration of cadmium was comparatively low. However, the concentration of metals was exceeding the RDA (Recommended Dietary Allowance of USA) limits. Hence, continuous fish consumption may create health problems for the consumers. The results of the present study are alarming and suggest implementing environmental laws and initiation of a biomonitoring program of the river.
Haniffa, Mohammed Abdul Kader; Sheela, Paul Asir Jeya; Kavitha, Kumaresan; Jais, Abdul Manan Mat
Murrel namely Channa striatus or haruan contains all essential elements to maintain good health and to recover the lost energy after prolonged illness. The fatty acid composition (% of total fatty acid) indicated the abundant presence of C16:0 fatty acid as 30% and the other major fatty acids were C22:6 (15%), C20:4 (19%), C18:1 (12%) and C18:0 (15%). Haruan contains arachidonic acid (C20:4) as 19.0%, a precursor for prostaglandin and thromboxane biosyntheses. Both fatty and amino acids are important components for wound healing processes. Both the fillet and mucus extracts of haruan were found to exhibit a concentration dependent antinociceptive activity. In vitro antioxidant activity was higher in Channa roe protein hydrolysate than in Labeo roe protein hydrolysate in both DPPH radical scavenging and ferric reducing power. Protein content of roe concentrates (RPC) was found to be 90.2% (Channa) and 82.5% (Lates). Water absorption, oil absorption, foam capacity, stability and emulsifying capacity were found to be higher in Channa RPC than in Lates RPC. Characterization of protein hydrolysates from muscle and myofibrillar samples of haruan showed different kinetic and proteolytic activities. The skin extract of haruan influences the serotonergic receptor system thus they can function as an anti-depressant. Thus, haruan is the best example for food as medicine.
Ahmad, Habib; Yousafzai, Ali Muhammad; Siraj, Muhammad; Ahmad, Rashid; Ahmad, Israr; Nadeem, Muhammad Shahid; Ahmad, Waqar; Akbar, Nazia; Muhammad, Khushi
The contamination of aquatic systems with heavy metals is affecting the fish population and hence results in a decline of productivity rate. River Kabul is a transcountry river originating at Paghman province in Afghanistan and inters in Khyber Pakhtunkhwa province of Pakistan and it is the major source of irrigation and more than 54 fish species have been reported in the river. Present study aimed at the estimation of heavy metals load in the fish living in River Kabul. Heavy metals including chromium, nickel, copper, zinc, cadmium, and lead were determined through atomic absorption spectrophotometer after tissue digestion by adopting standard procedures. Concentrations of these metals were recorded in muscles and liver of five native fish species, namely, Wallago attu, Aorichthys seenghala, Cyprinus carpio, Labeo dyocheilus, and Ompok bimaculatus. The concentrations of chromium, nickel, copper, zinc, and lead were higher in both of the tissues, whereas the concentration of cadmium was comparatively low. However, the concentration of metals was exceeding the RDA (Recommended Dietary Allowance of USA) limits. Hence, continuous fish consumption may create health problems for the consumers. The results of the present study are alarming and suggest implementing environmental laws and initiation of a biomonitoring program of the river. PMID:26339622
Patra, Avijit; Mondal, Anjan; Banerjee, Sayani; Adikesavalu, Harresh; Joardar, Siddhartha Narayan; Abraham, Thangapalam Jawahar
The present study characterized Argulus spp. infecting the cultured carps using 18S rRNA gene sequences, estimated the genetic similarity among Argulus spp. and established their phylogenetic relationship. Of the 320 fish samples screened, 34 fish (10.6%) had Argulus infection. The parasitic frequency index (PFI) was observed to be high (20%) in Hypophthalmichthys molitrix and Labeo bata. The frequency of infection was high in September (PFI: 17%) and October (PFI: 12.9%). The 18S rRNA sequences of five A. bengalensis (KF583878, KF192316, KM016968, KM016969, and KM016970) and one A. siamensis (KF583879) of this study showed genetic heterogeneity and exhibited 77-99% homology among the 18S rRNA gene sequences of Argulus spp. of NCBI GenBank database. Among the Indian Argulus spp. the sequence homology was 87–100%. Evolutionary pair-wise distances between Indian Argulus spp. and other Argulus spp. ranged from 0 to 20.20%. In the phylogenetic tree, all the crustaceans were clustered together as a separate clade with two distinct lineages. The lineage-1 comprised exclusive of Branchiura (Argulus spp.). All Argulus bengalensis clustered together and A. siamensis (KF583879) was closely related to Argulus sp. JN558648. The results of the present study provided baseline data for future work on population structure analysis of Indian Argulus species. PMID:28097169
Yang, Lei; Sado, Tetsuya; Vincent Hirt, M; Pasco-Viel, Emmanuel; Arunachalam, M; Li, Junbing; Wang, Xuzhen; Freyhof, Jörg; Saitoh, Kenji; Simons, Andrew M; Miya, Masaki; He, Shunping; Mayden, Richard L
Cyprininae is the largest subfamily (>1300 species) of the family Cyprinidae and contains more polyploid species (∼400) than any other group of fishes. We examined the phylogenetic relationships of the Cyprininae based on extensive taxon, geographical, and genomic sampling of the taxa, using both mitochondrial and nuclear genes to address the phylogenetic challenges posed by polyploidy. Four datasets were analyzed in this study: two mitochondrial gene datasets (465 and 791 taxa, 5604bp), a mitogenome dataset (85 taxa, 14,771bp), and a cloned nuclear RAG1 dataset (97 taxa, 1497bp). Based on resulting trees, the subfamily Cyprininae was subdivided into 11 tribes: Probarbini (new; Probarbus+Catlocarpio), Labeonini Bleeker, 1859 (Labeo & allies), Torini Karaman, 1971 (Tor, Labeobarbus & allies), Smiliogastrini Bleeker, 1863 (Puntius, Enteromius & allies), Poropuntiini (Poropuntius & allies), Cyprinini Rafinesque, 1815 (Cyprinus & allies), Acrossocheilini (new; Acrossocheilus & allies), Spinibarbini (new; Spinibarbus), Schizothoracini McClelland, 1842 (Schizothorax & allies), Schizopygopsini Mirza, 1991 (Schizopygopsis & allies), and Barbini Bleeker, 1859 (Barbus & allies). Phylogenetic relationships within each tribe were discussed. Two or three distinct RAG1 lineages were identified for each of the following tribes Torini, Cyprinini, Spinibarbini, and Barbini, indicating their hybrid origin. The hexaploid African Labeobarbus & allies and Western Asian Capoeta are likely derived from two independent hybridization events between their respective maternal tetraploid ancestors and Cyprinion.
Otero, Olga; Pinton, Aurélie; Mackaye, Hassan Taisso; Likius, Andossa; Vignaud, Patrick; Brunet, Michel
This is the first extensive study of a freshwater fish fauna from a Pliocene site in Central Africa, based on fossils collected at the KL2 site in the fossiliferous area of Kolle (Lower Pliocene, Chad). A relatively high fish diversity is revealed, confirming the presence of 19 taxa: Polypteriformes, Polypteridae ( Polypterus sp.); Osteoglossiformes, Osteoglossidae ( Heterotis sp.), Mormyriformes, Gymnarchidae ( Gymnarchus sp. cf. niloticus); Cypriniformes, Cyprinidae ( Labeo sp.); Characiformes, Alestidae ( Hydrocynus; Alestinae type Alestes/ Brycinus; Sindacharax sp. cf. deserti, Sindacharax sp.), Distichodontidae ( Distichodus sp.); Siluriformes, Ariidae (cf. Calarius), ?Bagridae (cf. Bagrus), Claroteidae (cf. Clarotes), Mochokidae ( Synodontis sp.), Clariidae ( Clarias sp. or Heterobranchus sp.); Perciformes family indet. ( Semlikiichthys sp. cf. darsao), Latidae ( Lates sp. cf. niloticus), Cichlidae indet., and Perciformes indet.; Tetraodontiformes Tetraodontidae ( Tetraodon sp.). The aquatic environment corresponding to the fossil fish assemblage might be a floodplain crossed by well-oxygenated open waters. Compared with a contemporaneous East African region, the mid-Pliocene Chadian fish diversity reveals a certain endemicity, while connections between the Niger and the Chadian basin are suspected because of the presence of a freshwater ariid fish in Kolle.
Rahman, M F; Jalal, K C A; Jahan, Nasrin; Kamaruzzaman, B Y; Ara, R; Arshad, A
Coordination among the different stakeholders at policy planning, implementation and target beneficiary level, particularly among the agencies responsible for development and management of water resources, agriculture and fisheries, is essential for overall sustainable development. Stocking of larger fingerlings at suitable stocking densities of endemic (rohu, catla, mrigal) and exotic (silver carp, bighead carp, common carp/mirror carp) species should be stocked at varying proportion. Floodplain fish production depends only on the natural fertility of the water bodies. Technological interventions should include the installation of low cost bamboo fencing at water inlet and outlet points and setting of ring culverts for maintaining suitable levels of water for fish culture without hampering the production of rice and other crops in the intervention areas, selective stocking with native and exotic carps, restricted fishing for certain period of time and guarding. It is expected to exert positive influences in enhancing the standing crop and biodiversity of non-stocked species of fishes in the intervention seasonal floodplain. Entry of fish larvae, hatchlings and young fry of wild non-stocked fishes into the seasonal floodplains because of large fence spacing (approximately 1.0 cm), could restrict fishing for certain period, undisturbed habitat and guarding could contribute to higher productivity and enhancement of fish biodiversity in the seasonal floodplains. Proper motivation and effective cooperation of the beneficiaries are extremely important to culture fish in the seasonal floodplains under community based management system. Institutional support and constant vigilance from the Department of Fisheries (DoF) and local administrations are indispensable to ensure the sustainability of fish culture initiatives in the seasonal floodplains. Active participation and involvement of the local community people in all stages of fish culture operation beginning from
Gownaris, Natasha J.; Pikitch, Ellen K.; Ojwang, William O.; Michener, Robert; Kaufman, Les
Background and Trophic Diversity Study Lake Turkana is an understudied desert lake shared by Kenya and Ethiopia. This system is at the precipice of large-scale changes in ecological function due to climate change and economic development along its major inflowing river, the Omo River. To anticipate response by the fish community to these changes, we quantified trophic diversity for seven ecological disparate species (Alestes baremose, Hydrocynus forskalli, Labeo horie, Lates niloticus, Oreochromis niloticus, Synodontis schall, and Tilapia zillii) using stable isotopes. Based on their marked morphological differentiation, we postulated that dietary niches of these species would be similar in size but show little overlap. The degree of trophic diversity varied greatly among the species studied, refuting our hypothesis regarding dietary niche size. Oreochromis niloticus and L. niloticus had the highest trophic diversity and significantly larger dietary niches than T. zillii, A. baremose and H. forskalli. Low overlap among the dietary niches of the seven species, with the exception of the synodontid catfish S. schall, is consistent with our second hypothesis. Predicting Species’ Vulnerability Breeding vulnerability was highest among those species with the lowest trophic diversity. We predict that in suffering two strikes against them, A. baremose, H. forskalli, T. zillii, and L. horie will be most affected by the highly altered Lake Turkana ecosystem and that O. niloticus, L. niloticus and S. schall will be least affected. Low vulnerability among O. niloticus and L. niloticus is promising for the future of the lake’s fishery, but the third most important fishery species (L. horie) will be highly vulnerable to impending ecosystem change. T. zillii should be treated as separate from O. niloticus in the fishery given higher sensitivity and a different ecological role. We see potential for expansion of the fishery for S. schall but don’t recommend the development of
Wiktelius, S; Edwards, C A
maximum and mean residues in the various groups of terrestrial and aquatic invertebrates and vertebrates. Overall, residues in the fauna were the greatest for DDT, followed in turn by those of dieldrin, HCH, endosulfan, and endrin, with small amounts of aldrin and toxaphene being found in some animals. There were relatively few reports of OCL residues in terrestrial invertebrates and virtually none in aquatic invertebrates. Only a few reports demonstrated OCL residues in terrestrial vertebrates, although high levels of DDT, dieldrin, and HCH were found in crocodile eggs and large residues of dieldrin occurred in bats, squirrels, and monkeys. Considerable OCL residues were reported in a few species of fish, especially Barbus, Clarias, Hydrocynus, Labeo, Sarotherodon, Epiplatys, and Synodontis. These residues were at levels that could have caused chronic toxicity or behavioral changes. The calculated maximum and mean OCL residues in the various elements of the African fauna until 1995 were compared with those calculated for corresponding faunal groups in Europe and the U.S. from their development and introduction up to 1973. The OCL residues reported in African fauna between 1971 and 1975 tended to be significantly higher overall than those published for Europe and the U.S. In particular, residues of DDT and dieldrin in African birds and their eggs were greater than those that had been incriminated as causing significant eggshell thinning and reproductive failure in European and U.S. aquatic and terrestrial birds up to 1973. Additionally, high DDT and dieldrin residues were reported from some species of African fish at levels that could potentially affect their reproduction, have chronic toxic and behavioral effects, and even drastically affect populations. Holistic case studies on the use of OCLs to control tsetse flies and desert locusts were discussed. OCL levels in trophic levels of fauna associated with Lake Kariba (between Zambia and Zimbabwe) were summarized. (AB