Information regarding molecular characteristics of leptin protein in different animal species in-cluding fish is scarce. With the aim of characterizing the native leptin protein of Indian major carprohu (Labeo rohita)...Information regarding molecular characteristics of leptin protein in different animal species in-cluding fish is scarce. With the aim of characterizing the native leptin protein of Indian major carprohu (Labeo rohita), at molecular level, the present study was designed to isolate rohu leptin from its hepatocytes (the prime source of leptin in fish) and immunobiochemical characterization of the same, subsequently. In the present study, chemical treatment and ultra-sonication technique was used for isolating leptin from rohu liver tissue. Purification of the protein was attempted using affinity column chromatography. The molecular, biophysical and serological characterization of rohu leptin was carried out by 2D-gel electrophoreis, SDS-PAGE, MALDI-TOF Mass spectroscopy and Western blot. The SDS-PAGE and 2D gel analysis revealed that rohu native leptin possesses molecular mass of 16 kDa. Western blot analysis showed that the fish hepatocytes possessed the sero-reactive leptin protein of 16 kDa. MALDI-TOF mass spectroscopy and peptide analysis showed the molecular mass of rohu leptin as 16283.38 Da. The serodiagnostic potential of native leptin of rohu was revealed for the first time while assessing its serological responses by ELISA using anti-leptin antibodies.展开更多
Unwarranted accumulation of halogenated compounds in the rivers and streams has in recent years emerged due to the widespread use agricultural pesticides. The presence of these halogenated compounds in the water does ...Unwarranted accumulation of halogenated compounds in the rivers and streams has in recent years emerged due to the widespread use agricultural pesticides. The presence of these halogenated compounds in the water does not only suppress the immune system of fish but adversely induce serious morbidity and mortality among cultured stocks. Importantly, gradual accumulation of these compounds in the system of cultured and wild freshwater fish species cultured in ponds and floating net-cages in dams and rivers, respectively, poses some risks to humans, the end users. In this study, we attempted to isolate bacteria from the gut of pond-reared rohu (Labeo rohita) in Myanmar, screened the isolated bacteria for dehalogenase gene using molecular technique and tested the ability of these bacteria to degrade halogenated compounds in vitro. The eight bacterial strains studied were identified as Enterobacter mori strain MK- 121001, Enterobacter cloacae strains MK121003, MK-121004, MK121010, Ralstonia solanacearum strain 121002, Acinetobacter baumannii strain MK121007, Chromobacterium violaceum strain MK121009 and Pantoea vagans strain 121011. Only three bacterial strains (MK121002, MK121007 and MK121009) were capable of degrading 2,2-dichloropropionic acid (2,2-DCP) as the sole carbon source up to a final substrate concentration of 20 mM. Their mean growth doubling time ranging from 6-23 hours with the maximum of chloride ion released of 85%. PCR amplifica- tion with oligonucleotide primers designed from group I dehalogenase revealed the presence of deha- logenase genes in all isolates suggesting dehalogenase gene in strains 121001, 121003, 121004, 121010 and 121011 were silenced. In contrast, group II dehalogenase primers did not show any PCR amplification. These results suggest that MK121002, MK121007 and MK121009 only encode a group I dehalogenase and its non-stereoselectivity is in agreement with previoulsly described group I haloacid dehalogenase. The partial gene sequences were blasted but no significant sequence identity was observed. Therefore, it suggest the 2-haloacid dehalogenase of MK121002, MK12-1007 and MK121009 might be a novel group I 2-haloacid dehalogenase. The results indicated a broad distribution of dehalogenation genes in many micro- bial genomes that harbor dehalogenase(s) due to the exposure of the microorganisms to the naturally occurring or man-made halogenated compounds in the environmental systems. So far, microorganisms capable of producing dehalogenases were mainly isolated from soil and scarcely from aquatic animals and their environments. To the authors’ knowledge, this is the first report on the isolation of dehalogenase-producing bacteria from the gut of pond-reared freshwater fish, Labeo rohita, in Myanmar.展开更多
Multi-strain probiotics provide the most dependable approaches to improve health,immune response,and disease resistance in farmed fishes.In the present study,we examined the effects of multi-species probiotics on the ...Multi-strain probiotics provide the most dependable approaches to improve health,immune response,and disease resistance in farmed fishes.In the present study,we examined the effects of multi-species probiotics on the survival,growth,immune response,and disease resistance of rohu(Labeo rohita)larvae.Newly hatched larvae from the day of first feeding(average weight of 0.003 g)were reared with multi-species probiotics having a combination of Bacillus subtilis(109 colony forming units(cfu)/mL)and Lactobacillus spp.(Lactobacillus plantarum,Lactobacillus buchneri-1011 cfu/mL)in water containing doses of 0(control-C),0.5 mL/L(treatment 1-T1),and 1.0 mL/L(treatment 2-T2)in triplicates for 90 days.After the experiment,a challenge test was performed to assess the fish's resistance to pathogenic Aeromonas veronii.Significantly higher survival was recorded in larvae of treated groups(87%in T2 and 79%in T1)compared to the control(62%).Significantly higher growth performance(weight gain and specific growth rate—SGR)was shown by the probiotic-treated larval groups compared to the control.Probiotic supplementation resulted in significantly higher counts of total viable colony(TVC)and lactic acid bacteria(LAB)in the intestine.Some immunological parameters(mucosal fold fattening,goblet cell abundance,expansion of lamina propria and enterocytes)of the gut were significantly better in probiotic-treated fish.The liver of treated fish showed irregular shape nuclei turning into regular shape and reducing spaces between the hepatic cells.Probiotic-treated fish had the highest post-challenge survival rate(90%)against A.veronii infection.The erythrocytes of challenged fish treated with probiotics had significantly lower frequencies of various nuclear and cellular abnormalities.These findings suggest that multi-species probiotic supplements could improve the survival,growth,health status,and immune response of rohu in the early stages of its development.展开更多
Background: The homeobox containing transcription factor Nanog plays crucial roles in embryonic development/proliferation and/or maintenance of spermatogonial stem cells(SSCs) via interacting with transcription factor...Background: The homeobox containing transcription factor Nanog plays crucial roles in embryonic development/proliferation and/or maintenance of spermatogonial stem cells(SSCs) via interacting with transcription factors such as Oct4 and Sox2 in mammals. However, knowledge of its exact mechanistic pathways remains unexploited. Very little is known about teleost Nanog. Information on the Nanog gene of farmed rohu carp(Labeo rohita) is lacking. We cloned and characterized the Nanog gene of rohu carp to understand the expression pattern in early developmental stages and also deduced the genomic organization including promoter elements.Results: Rohu Nanog(LrNanog) cDNA comprised an open reading frame of 1,161 nucleotides bearing a structural homeodomain; whereas, the genomic structure contained four exons and three introns suggesting that it is homologous to mammalian counterparts. Phylogenetical y, it was closely related to freshwater counterparts. Protein sequence(386 AA of42.65 kDa) comparison revealed its low similarity with other vertebrate counterparts except that of the conserved homeodomain. Tissue distribution analysis revealed the existence of LrNanog transcripts only in adult gonads. The heightened abundances in the ovary and proliferating spermatogonia suggested its participations in maternal inheritance and male germ cell development. The potentiating abundances from fertilized egg onwards peaking at blastula stage vis-à-vis decreasing levels from gastrula stage onwards demonstrated its role in embryonic stem cell development. We also provided evidence of its presence in SSCs by western blotting analysis. Further, the promoter region was characterized, predicting a basal core promoter and other consensus elements.Conclusion: The molecular characterization of LrNanog and its documented expression profiling at transcript and protein levels are indicative of its functional linkage with embryonic/spermatogonial stem cell maintenance. This is the first report of LrNanog genomic organization including its promoter sequence information with predicted regulatory elements of a large-bodied carp species. This will be useful for elucidating its mechanism expression in future. Nanog could be used as a potential biomarker for proliferating carp SSCs.展开更多
The gel-forming ability of rohu (Labeo rohita) mince with and without egg white powder (EW) was investigated. Gel from washed mince (washed gel) was prepared under two setting conditions: kamaboko (40°C) and modo...The gel-forming ability of rohu (Labeo rohita) mince with and without egg white powder (EW) was investigated. Gel from washed mince (washed gel) was prepared under two setting conditions: kamaboko (40°C) and modori (60°C). The gel-forming ability of kamaboko and modori gels was improved by the addition of EW at 2%. The autolytic inhibition of kamaboko gel was obtained in gel added with 2% EW, and 1% EW of modori gel. No marked change was observed in the TCA-soluble peptide content of either gel with the addition of EW above 1%. No effect on the whiteness of both gels was shown after the addition of EW. The addition of EW exhibited smaller cavities and a more compact fibrous network in microstructure.展开更多
Fry rearing is one of the important stage which aims at obtaining high growth and survival for production of fingerlings required for stocking into grow out ponds as well as rehabilitation in natural habitat. This exp...Fry rearing is one of the important stage which aims at obtaining high growth and survival for production of fingerlings required for stocking into grow out ponds as well as rehabilitation in natural habitat. This experiment was conducted with the purpose to test the effect of dietary crude protein level (CP %) of feeds prepared from similar feed ingredients in different ratios on growth performance and survival rate of Rohu fry (Labeorohita). An initial density of 100 fry/m2 was maintained in hapa fixed in the cemented tank. The dietary CP% level of feed tested were 20% CP, 25% CP, 30% CP, and 35% CP fed at 5% body weight. The experiment ran for 53 nursing days. The results showed that there was no significant difference (P > 0.05) in the growth rate (g/day) of fry among treatments. Rather group fed with higher protein level grew comparatively better indicating possibility of increasing need of protein in diets. However, the survival rate (%) of rohu fry was significantly different (P < 0.05) in each tested CP% level of feed. Highest survival (82%) of fry was found in the feed of CP 35% and lowest (56%) in the feed of CP 25%. It was predicted that feed with increasing level of CP % in diet is essential for increasing survival rate.展开更多
The present work reports the length–weight relationships(LWR)and condition factor relationships for Labeo calbasu collected from Upstream(Matyari)Kotri barrage at,River Indus,Pakistan,because stock assessment helps t...The present work reports the length–weight relationships(LWR)and condition factor relationships for Labeo calbasu collected from Upstream(Matyari)Kotri barrage at,River Indus,Pakistan,because stock assessment helps the fisheries managers to conserve the commercially important fish.Morphological characters of fish as well as Length-weight relationship are an important tool for fishery management.The results of LWR(W=aLb),for L calbasu.Representing negative allometric growth pattern.LWRs and condition factor relationships were found significantly correlated.A total of 200 and 190 specimens from upstream and downstream were collected,respectively.The assessed values of length-weight correlation and condition factor were calculated as Kn=39.663(LeCren),and K=11.915(Fulton)for upstream and Kn=44.066 and K=13.872 for downstream.Length-weight was found with a strong correlation of n=2.892,a=0.0235 with r2=0.934 for upstream population then the downstream population.The results of this work would be beneficial for sustainable management as well as fishery managers.展开更多
文摘Information regarding molecular characteristics of leptin protein in different animal species in-cluding fish is scarce. With the aim of characterizing the native leptin protein of Indian major carprohu (Labeo rohita), at molecular level, the present study was designed to isolate rohu leptin from its hepatocytes (the prime source of leptin in fish) and immunobiochemical characterization of the same, subsequently. In the present study, chemical treatment and ultra-sonication technique was used for isolating leptin from rohu liver tissue. Purification of the protein was attempted using affinity column chromatography. The molecular, biophysical and serological characterization of rohu leptin was carried out by 2D-gel electrophoreis, SDS-PAGE, MALDI-TOF Mass spectroscopy and Western blot. The SDS-PAGE and 2D gel analysis revealed that rohu native leptin possesses molecular mass of 16 kDa. Western blot analysis showed that the fish hepatocytes possessed the sero-reactive leptin protein of 16 kDa. MALDI-TOF mass spectroscopy and peptide analysis showed the molecular mass of rohu leptin as 16283.38 Da. The serodiagnostic potential of native leptin of rohu was revealed for the first time while assessing its serological responses by ELISA using anti-leptin antibodies.
文摘Unwarranted accumulation of halogenated compounds in the rivers and streams has in recent years emerged due to the widespread use agricultural pesticides. The presence of these halogenated compounds in the water does not only suppress the immune system of fish but adversely induce serious morbidity and mortality among cultured stocks. Importantly, gradual accumulation of these compounds in the system of cultured and wild freshwater fish species cultured in ponds and floating net-cages in dams and rivers, respectively, poses some risks to humans, the end users. In this study, we attempted to isolate bacteria from the gut of pond-reared rohu (Labeo rohita) in Myanmar, screened the isolated bacteria for dehalogenase gene using molecular technique and tested the ability of these bacteria to degrade halogenated compounds in vitro. The eight bacterial strains studied were identified as Enterobacter mori strain MK- 121001, Enterobacter cloacae strains MK121003, MK-121004, MK121010, Ralstonia solanacearum strain 121002, Acinetobacter baumannii strain MK121007, Chromobacterium violaceum strain MK121009 and Pantoea vagans strain 121011. Only three bacterial strains (MK121002, MK121007 and MK121009) were capable of degrading 2,2-dichloropropionic acid (2,2-DCP) as the sole carbon source up to a final substrate concentration of 20 mM. Their mean growth doubling time ranging from 6-23 hours with the maximum of chloride ion released of 85%. PCR amplifica- tion with oligonucleotide primers designed from group I dehalogenase revealed the presence of deha- logenase genes in all isolates suggesting dehalogenase gene in strains 121001, 121003, 121004, 121010 and 121011 were silenced. In contrast, group II dehalogenase primers did not show any PCR amplification. These results suggest that MK121002, MK121007 and MK121009 only encode a group I dehalogenase and its non-stereoselectivity is in agreement with previoulsly described group I haloacid dehalogenase. The partial gene sequences were blasted but no significant sequence identity was observed. Therefore, it suggest the 2-haloacid dehalogenase of MK121002, MK12-1007 and MK121009 might be a novel group I 2-haloacid dehalogenase. The results indicated a broad distribution of dehalogenation genes in many micro- bial genomes that harbor dehalogenase(s) due to the exposure of the microorganisms to the naturally occurring or man-made halogenated compounds in the environmental systems. So far, microorganisms capable of producing dehalogenases were mainly isolated from soil and scarcely from aquatic animals and their environments. To the authors’ knowledge, this is the first report on the isolation of dehalogenase-producing bacteria from the gut of pond-reared freshwater fish, Labeo rohita, in Myanmar.
基金Krishi Gobeshona Foundation,Bangladesh Agricultural Research Council,Dhaka,supported a grant(CGP TF 75-F/20)the corresponding author.Bangladesh Agricultural University Research System(BAURES)is gratefully acknowledged for the financial management of this project(Project No.2020/954/KGF).
文摘Multi-strain probiotics provide the most dependable approaches to improve health,immune response,and disease resistance in farmed fishes.In the present study,we examined the effects of multi-species probiotics on the survival,growth,immune response,and disease resistance of rohu(Labeo rohita)larvae.Newly hatched larvae from the day of first feeding(average weight of 0.003 g)were reared with multi-species probiotics having a combination of Bacillus subtilis(109 colony forming units(cfu)/mL)and Lactobacillus spp.(Lactobacillus plantarum,Lactobacillus buchneri-1011 cfu/mL)in water containing doses of 0(control-C),0.5 mL/L(treatment 1-T1),and 1.0 mL/L(treatment 2-T2)in triplicates for 90 days.After the experiment,a challenge test was performed to assess the fish's resistance to pathogenic Aeromonas veronii.Significantly higher survival was recorded in larvae of treated groups(87%in T2 and 79%in T1)compared to the control(62%).Significantly higher growth performance(weight gain and specific growth rate—SGR)was shown by the probiotic-treated larval groups compared to the control.Probiotic supplementation resulted in significantly higher counts of total viable colony(TVC)and lactic acid bacteria(LAB)in the intestine.Some immunological parameters(mucosal fold fattening,goblet cell abundance,expansion of lamina propria and enterocytes)of the gut were significantly better in probiotic-treated fish.The liver of treated fish showed irregular shape nuclei turning into regular shape and reducing spaces between the hepatic cells.Probiotic-treated fish had the highest post-challenge survival rate(90%)against A.veronii infection.The erythrocytes of challenged fish treated with probiotics had significantly lower frequencies of various nuclear and cellular abnormalities.These findings suggest that multi-species probiotic supplements could improve the survival,growth,health status,and immune response of rohu in the early stages of its development.
基金provided by grant from Indian Council of Agricultural Research (ICAR) and Department of Biotechnology(DBT),Government of India
文摘Background: The homeobox containing transcription factor Nanog plays crucial roles in embryonic development/proliferation and/or maintenance of spermatogonial stem cells(SSCs) via interacting with transcription factors such as Oct4 and Sox2 in mammals. However, knowledge of its exact mechanistic pathways remains unexploited. Very little is known about teleost Nanog. Information on the Nanog gene of farmed rohu carp(Labeo rohita) is lacking. We cloned and characterized the Nanog gene of rohu carp to understand the expression pattern in early developmental stages and also deduced the genomic organization including promoter elements.Results: Rohu Nanog(LrNanog) cDNA comprised an open reading frame of 1,161 nucleotides bearing a structural homeodomain; whereas, the genomic structure contained four exons and three introns suggesting that it is homologous to mammalian counterparts. Phylogenetical y, it was closely related to freshwater counterparts. Protein sequence(386 AA of42.65 kDa) comparison revealed its low similarity with other vertebrate counterparts except that of the conserved homeodomain. Tissue distribution analysis revealed the existence of LrNanog transcripts only in adult gonads. The heightened abundances in the ovary and proliferating spermatogonia suggested its participations in maternal inheritance and male germ cell development. The potentiating abundances from fertilized egg onwards peaking at blastula stage vis-à-vis decreasing levels from gastrula stage onwards demonstrated its role in embryonic stem cell development. We also provided evidence of its presence in SSCs by western blotting analysis. Further, the promoter region was characterized, predicting a basal core promoter and other consensus elements.Conclusion: The molecular characterization of LrNanog and its documented expression profiling at transcript and protein levels are indicative of its functional linkage with embryonic/spermatogonial stem cell maintenance. This is the first report of LrNanog genomic organization including its promoter sequence information with predicted regulatory elements of a large-bodied carp species. This will be useful for elucidating its mechanism expression in future. Nanog could be used as a potential biomarker for proliferating carp SSCs.
文摘The gel-forming ability of rohu (Labeo rohita) mince with and without egg white powder (EW) was investigated. Gel from washed mince (washed gel) was prepared under two setting conditions: kamaboko (40°C) and modori (60°C). The gel-forming ability of kamaboko and modori gels was improved by the addition of EW at 2%. The autolytic inhibition of kamaboko gel was obtained in gel added with 2% EW, and 1% EW of modori gel. No marked change was observed in the TCA-soluble peptide content of either gel with the addition of EW above 1%. No effect on the whiteness of both gels was shown after the addition of EW. The addition of EW exhibited smaller cavities and a more compact fibrous network in microstructure.
文摘Fry rearing is one of the important stage which aims at obtaining high growth and survival for production of fingerlings required for stocking into grow out ponds as well as rehabilitation in natural habitat. This experiment was conducted with the purpose to test the effect of dietary crude protein level (CP %) of feeds prepared from similar feed ingredients in different ratios on growth performance and survival rate of Rohu fry (Labeorohita). An initial density of 100 fry/m2 was maintained in hapa fixed in the cemented tank. The dietary CP% level of feed tested were 20% CP, 25% CP, 30% CP, and 35% CP fed at 5% body weight. The experiment ran for 53 nursing days. The results showed that there was no significant difference (P > 0.05) in the growth rate (g/day) of fry among treatments. Rather group fed with higher protein level grew comparatively better indicating possibility of increasing need of protein in diets. However, the survival rate (%) of rohu fry was significantly different (P < 0.05) in each tested CP% level of feed. Highest survival (82%) of fry was found in the feed of CP 35% and lowest (56%) in the feed of CP 25%. It was predicted that feed with increasing level of CP % in diet is essential for increasing survival rate.
基金The present work is partial work of M.Phil research scholar.
文摘The present work reports the length–weight relationships(LWR)and condition factor relationships for Labeo calbasu collected from Upstream(Matyari)Kotri barrage at,River Indus,Pakistan,because stock assessment helps the fisheries managers to conserve the commercially important fish.Morphological characters of fish as well as Length-weight relationship are an important tool for fishery management.The results of LWR(W=aLb),for L calbasu.Representing negative allometric growth pattern.LWRs and condition factor relationships were found significantly correlated.A total of 200 and 190 specimens from upstream and downstream were collected,respectively.The assessed values of length-weight correlation and condition factor were calculated as Kn=39.663(LeCren),and K=11.915(Fulton)for upstream and Kn=44.066 and K=13.872 for downstream.Length-weight was found with a strong correlation of n=2.892,a=0.0235 with r2=0.934 for upstream population then the downstream population.The results of this work would be beneficial for sustainable management as well as fishery managers.
