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1.
Genomics ; 2020 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-32531445

RESUMO

The Queen loach (Botia dario), an ornamental fish species having export potential, belongs to family Cobitidae of order Cypriniformes. The dull colouration in captive condition as compared to nature is a drawback in ornamental fisheries. We report the first comparative transcriptomic analysis of Cultured (CBD) and Natural (NBD) B. dario using bioinformatics tools. Total 26 and 7 key genes for melanin and carotenoid colouration were found, respectively. KEGG pathway annotations of the genes were carried out, to annotate and describe their relevance for pigmentation. The qPCR validation of genes confirmed their expression pattern in the skin and muscle. Differential expression of, slc7a11, asip1, mc1r, dct, tyrp1a, tyr, bcdo2, csf1r, plin2, gsta2, star3 and stard5 in the skin and muscle tissues revealed the reasons for wild versus cultured colour variation. The molecular data was further supported by low yellowness and redness values of CBD skin and muscle in a colorimeter.

2.
Gene ; 754: 144860, 2020 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-32531457

RESUMO

Arunachal Pradesh, the largest state of North-East India covers almost 60.93% of the Eastern Himalayan hotspot. Fish diversity and species identification is utmost important for fisheries management. But, in some cases morphological characteristics based identification is difficult for a non-specialist to perform. In view of the above, the present study emphasized on the assessment of DNA barcoding, phylogenetics and genetic diversity of fish species in the Ranganadi River, Arunachal Pradesh, India. India. Arunachal Pradesh, the largest state of North-East India covers almost 60.93% of the Eastern Himalayan hotspot. Altogether 114 specimens, representing 22 species, belonging to 3 orders and 5 families were successfully barcoded and found to be 98-100% identical from both GenBank and BOLD databases. Out of these 22 fish species, it was found that one species assessed was Endangered, three species as Near Threatened and one species as Vulnerable. A Neighbour Joining (NJ) tree was constructed using Rstudio for the purpose of a phylogenetic analysis of the identified species. The barcoding gap analysis using K2P, P-distance and Jukes-Cantor was done to detect the presence of cryptic species and barcoding success. The nucleotide base composition and genetic distance analysis were also performed, using MEGA 6.0. DNA Sequence Polymorphism v6.12.03 analysis revealed the nucleotide diversity (p) and haplotype diversity (Hd). The Hd for the whole dataset was found to be 0.975, which showed high genetic diversity in the Ranganadi River. Both morphological key identifying characters and molecular data corroborated the phylogenetic analysis. This COI barcode library, generated in the present study, not only helped in species identification and molecular study, but also in cryptic species identification.

3.
Zootaxa ; 4648(3): zootaxa.4648.3.6, 2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31716938

RESUMO

Mystus prabini, new species, is described from the Sinkin and the Dibang River in the Lower Dibang Valley District of Arunachal Pradesh, India. The new species differs from all South-Asian congeners except M. bleekeri, M. cavasius, M. zeylanicus,  M. falcarius, M. seengtee, M. cineraceus, M. ngasep, M. rufescens and M. ankutta in having a long adipose fin that reaches anteriorly (vs. distinctly does not reach) the base of the last dorsal-fin ray. The new species can be distinguished from the named nine species in having (vs. lacking) a narrow black mid-lateral stripe extending from the anterior region of tympanic spot to the rounded black spot at the caudal-fin base. The analysis of mitochondrial cytochrome c oxidase subunit I (COI) gene sequence shows that the K2P value between Mystus prabini and all other Mystus species ranges from 8.6-22.1%. Mystus prabini is closest genetically to M. bleekeri and M. albolineatus, from which species it has a genetic distance of 8.6% and 13.9%, respectively. The genetic distance (K2P) between the new species and M. dibrugarensis is 21.1%.


Assuntos
Peixes-Gato , Animais , Índia , Rios
4.
Mol Biol Rep ; 46(6): 5931-5939, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31401780

RESUMO

The immunity and health status of ornamental fish is an important aspect, as they are kept in a confined environment and various stressful conditions which lead to depletion of overall colourful appearance and mortality. The carotenoids can act as immunity boosters in captive aquarium system and may be supplemented in the feed as aquarium fish have no access to natural carotenoids. The study aimed to assess the role of carotenoid on the immunity of B. dario. Marigold petal meal is an important source of carotenoids and used in experimental diets. Four immunogenes namely IL20, TLR9, TRAIL, and Nramp in B. dario were characterized and also studied for their relative expression in the kidney after feeding the fish with marigold petal meal supplemented diet. The expression pattern of the genes was compared with the fish of nature. The IL20 and Nramp gene were upregulated significantly (p < 0.05) in the fish of nature as compared to the experimental fish at the 60th day of feeding carotenoid-rich diet. But the TLR9 and TRAIL gene was upregulated significantly (p < 0.05) in experimental fish as compared to nature. The haematological parameters of fish after feeding with the experimental enriched diet for 60 days also confirmed the role of carotenoids in immunity.


Assuntos
Carotenoides/metabolismo , Cipriniformes/imunologia , Ração Animal , Animais , Asteraceae , Dieta , Suplementos Nutricionais
5.
Sci Rep ; 9(1): 12585, 2019 08 29.
Artigo em Inglês | MEDLINE | ID: mdl-31467347

RESUMO

The genetic basis and expression patterns of key genes are important aspects of study to understand the colouration. This trait differs between wild and domesticated fish which is a matter of research. Botia dario is an indigenous fish, having ornamental and aesthetic value, which shows faded appearance in terms of colour in domesticated condition than wild. In the present study the carotenoid-fed B. dario were examined through incorporation of marigold petal meal in the diets at the rate of 5, 10 and 15% w/w along with wild fish. The carotenoid content of tissues that is skin, muscle and intestine along with intensity of colouration increased in a dose dependant manner of carotenoid in the diet. Important carotenoid-based colouration genes that is csf1r, BCDO2, SR-B1, MLN64, STAR5, GSTA2 and PLIN2 were characterized in the fish, to find out their role in fish pigmentation. The significant difference (p < 0.05) in the expression of these genes in different tissues, when compared among carotenoid-fed domesticated and wild fish, revealed the mechanism responsible for faded colouration and also revealed the means to enhance colour in the fish.

6.
Gene ; 713: 143967, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31279710

RESUMO

Phylogenetic tree using mitochondrial genes and nuclear genes have long been used for augmenting biological classification and understanding evolutionary processes in different lineage of life. But a basic question still exists for finding the most suitable gene for constructing robust phylogenetic tree. Much of the controversy appears due to monophyletic, paraphyletic and polyphyletic clade making deviations from original taxonomy. In the present study we report the first complete mitochondrial genome (mitogenome) of queen loach, generated through next-generation sequencing methods. The assembled mitogenome is a 16,492 bp circular DNA, comprising of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. Further in this study we also investigated the suitability of different mitochondrial region for phylogenetic analysis in Cyprinidae and loach group. For this genetic tree were constructed on COI, COII, COIII, 16S rRNA, 12S rRNA, Cyt b, ATPase 6, D-loop, ND1, ND2, ND3, ND4, ND5, and ND6 along with complete mitogenome. The complete mitogenome based phylogenetic tree got inclusive support from available classical taxonomy for these groups. On individual gene basis Cyt b, 12S rRNA, ND2 and ND3 also produced perfect clade at family and subfamily level. For rest of the genes polyphyly were observed for the fishes belonging to same family or subfamily which makes their use questionable for phylogenetic tree construction.


Assuntos
Cipriniformes/genética , Genes Mitocondriais , Genoma Mitocondrial , Mitocôndrias/genética , Filogenia , Animais , Análise de Sequência de DNA
7.
J Fish Dis ; 41(10): 1539-1548, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30105863

RESUMO

This study evaluated the effect of sublethal doses of antifungal drug miconazole nitrate (MCZ) on immunological responses and its role as a prophylactic drug against S. parasitica in Labeo rohita fingerlings. Fish were fed with sublethal doses of MCZ, that is, T1-6.30 mgMCZ kgBW-1 , T2-12.61 mgMCZ kgBW-1 and T3-25.22 mgMCZ kgBW-1 , and sampling was done at different time intervals for 240 hr. Immunological parameters viz. lysozyme activity, oxygen radical production and plasma antiprotease activity showed significant enhancement (p < 0.05) in fish fed with T2 and T3 doses. Expression of immune-relevant genes such as TLR-22 and ß2-M showed significantly higher expression at 6 hr and 24 hr of sampling in both liver and head kidney. However, these genes showed a downregulation after 120 hr of sampling in both the tissues. Preventive efficacy study showed that single dose of MCZ provides protection against oomycetes up to the fourth day of infection. Significantly higher mortality was observed in control diet-fed fish as compared to fish fed with MCZ medicated diet. Thus, it can be concluded that the MCZ can act as a potent antifungal agent for preventing oomycetes infection as well as to enhance the immune response.


Assuntos
Antiparasitários/administração & dosagem , Cyprinidae/imunologia , Cyprinidae/parasitologia , Doenças dos Peixes/prevenção & controle , Miconazol/administração & dosagem , Saprolegnia/efeitos dos fármacos , Fatores Etários , Ração Animal , Animais , Suplementos Nutricionais , Doenças dos Peixes/mortalidade , Doenças dos Peixes/parasitologia , Infecções/parasitologia , Muramidase/efeitos dos fármacos , Muramidase/metabolismo , Inibidores de Proteases/metabolismo , Espécies Reativas de Oxigênio/metabolismo
8.
J Anat ; 233(2): 177-192, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29806093

RESUMO

Aquaporin-mediated fluid transport in the mammalian efferent duct and epididymis is believed to play a role in sperm maturation and concentration. In fish, such as the marine teleost gilthead seabream (Sparus aurata), the control of fluid homeostasis in the spermatic duct seems also to be crucial for male fertility, but no information exists on the expression and distribution of aquaporins. In this study, reverse transcriptase-polymerase chain reaction and immunoblotting analyses, employing available and newly raised paralog-specific antibodies for seabream aquaporins, indicate that up to nine functional aquaporins, Aqp0a, -1aa, -1ab, -3a, -4a, -7, -8bb, -9b and -10b, are expressed in the spermatic duct. Immunolocalization of the channels in the resting spermatic duct reveals that Aqp0a, -1aa, -4a, -7 and -10b are expressed in the monolayered luminal epithelium, Aqp8b and -9b in smooth muscle fibers, and Aqp1ab and -3a in different interstitial lamina cells. In the epithelial cells, Aqp0a and -1aa are localized in the short apical microvilli, and Aqp4a and -10b show apical and basolateral staining, whereas Aqp7 is solely detected in vesicular compartments. Upon spermiation, an elongation of the epithelial cells sterocilia, as well as the folding of the epithelium, is observed. At this stage, single- and double-immunostaining, using two aquaporin paralogs or the Na+ /K+ -ATPase membrane marker, indicate that Aqp1ab, -3a, -7, -8bb and -9b staining remains unchanged, whereas in epithelial cells Aqp1aa translation is supressed, Aqp4a internalizes, and Aqp0a and -10b accumulate in the apical, lateral and basal plasma membrane. These findings uncover a cell type- and region-specific distribution of multiple aquaporins in the piscine spermatic duct, which shares conserved features of the mammalian system. The data therefore suggest that aquaporins may play different roles in the regulation of fluid homeostasis and sperm maturation in the male reproductive tract of fish.


Assuntos
Aquaporinas/metabolismo , Dourada/metabolismo , Cordão Espermático/metabolismo , Animais , Cílios/fisiologia , Células Epiteliais/fisiologia , Homeostase , Masculino
9.
Vet Immunol Immunopathol ; 188: 48-58, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28615127

RESUMO

This study investigates the effects of dietary lipopolysaccharide (LPS) as an immunostimulant on hematology, innate immunity, immune gene expression and protection against Edwardsiella tarda on Labeo bata. A basal diet supplemented with 0, 50, 100 and 150mg LPS kg-1diet was fed to the four different groups for 30days. The haematological (total erythrocyte count, total leukocyte count, total serum protein, albumin and globulin), innate immune parameters (respiratory burst, serum lysozyme, myeloperoxidase and serum bactericidal activity), immune gene expression (C3, ß-2 microglobulin, lysozyme g, transferrin, IFN-1, IFN-γ) were monitored at 7th, 15th, 30th day and one day post challenge (DPC) with E. tarda. All the studied haematological, innate immune parameters and expression of immune gene increased significantly (p≤0.05) in LPS fed group in comparison with control. However the group fed 100mgkg-1 LPS in feed showed highest activity on 7th day and 1DPC. The group fed 100mgkg-1 LPS also recorded highest relative percent survivability after challenge with E. tarda. Therefore this study suggests that LPS at 100mgkg-1 could be used as an immunostimulant in feed to enhance the protection of bata during periods of increased disease risk.


Assuntos
Adjuvantes Imunológicos/farmacologia , Carpas/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Imunidade/imunologia
10.
Fish Shellfish Immunol ; 66: 35-42, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28476667

RESUMO

The present study evaluated the paraprobiotic effect of heat-inactivated Bacillus amyloliquefaciens FPTB16 on immunological response and immune relevant gene expression in catla (Catla catla). Heat inactivation of viable cells of B. amyloliquefaciens was done at 60 °C for 2 h. For preparation of paraprobiotic supplemented diet, the heat-inactivated bacteria were added to the basal diet (control) at three different inclusion levels i.e., 107, 108 and 109 cells g-1 diet. Fish (25.98 ± 2.57 g) were fed with these diets and various immune responses and immune relevant gene expressions were measured after 4 weeks of feeding. Biochemical parameters were also measured along with the immunological responses. Immunological parameters viz. oxygen radical production, serum lysozyme activity and total serum protein content showed significant enhancement (p < 0.05) in fish fed with 108 and 109 cells g-1 diet. Significant enhancement in myeloperoxidase activity was observed in all the dietary groups compared to control. Alkaline phosphatase activity showed significant enhancement (p < 0.05) in fish fed with 107 and 108 cells g-1 diet. Biochemical parameters viz. GPT, GOT and glucose content did not show any significant difference in any of the dietary groups. Immune relevant genes viz. IL-1ß, TNF-α, C3 and iNOS showed significantly higher expression in either liver or head-kidney tissues in most of the cases. However, IFN-γ expression showed a down-regulation pattern in both the tissues. In conclusion, the dietary supplementation of heat-inactivated B. amyloliquefaciens enhanced the immunity of catla, particularly at 108 cells g-1 diet. The results collectively suggest the paraprobiotic applicability of B. amyloliquefaciens in aquaculture.


Assuntos
Bacillus amyloliquefaciens/química , Cyprinidae/genética , Cyprinidae/imunologia , Regulação da Expressão Gênica , Imunidade Inata , Probióticos/administração & dosagem , Ração Animal/análise , Animais , Cyprinidae/metabolismo , Dieta/veterinária , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Distribuição Aleatória , Reação em Cadeia da Polimerase em Tempo Real/veterinária
11.
Artigo em Inglês | MEDLINE | ID: mdl-28095297

RESUMO

Studies in teleosts suggest that progestins have crucial functions during early spermatogenesis. However, the role of the different progestin receptors in these mechanisms is poorly understood. In this work, we investigated the expression pattern and hormonal regulation of the classical nuclear progestin receptor (Pgr) in the gilthead seabream at three different stages of spermatogenesis: the resting (postspawning) phase, onset of spermatogenesis, and spermiation. Immunolocalization experiments using a seabream specific Pgr antibody revealed that the receptor was expressed in Sertoli and Leydig cells, and also in a subset of spermatogonia type A, throughout spermatogenesis. Short-term treatment of testis explants with 17ß-estradiol (E2) increased pgr mRNA expression at all stages, while the progestin 17α,20ß-dihydroxy-4-pregnen-3-one (17,20ßP) had the opposite effect. At the resting stage, Sertoli cell Pgr expression was positively correlated with the occurrence of proliferating spermatogonia type A in the tubules, and both processes were incremented in vitro by E2 likely through the estrogen receptor alpha (Era) expressed in Sertoli and Leydig cells. In contrast, treatment with 17,20ßP downregulated Pgr expression in somatic cells. The androgen 11-ketotestosterone (11-KT) upregulated pgr expression in Leydig cells and promoted the proliferation of mostly spermatogonia type B, but only during spermiation. No relationship between the changes in the cell type-specific expression of the Pgr with the entry into meiosis of germ cells was found. These data suggest a differential steroid regulation of Pgr expression during seabream spermatogenesis and the potential interplay of the E2/Era and 17,20ßP/Pgr pathways for the maintenance of spermatogonial renewal rather than entry into meiosis.


Assuntos
Núcleo Celular/metabolismo , Estradiol/metabolismo , Receptores de Progesterona/agonistas , Dourada/fisiologia , Espermatogênese , Espermatogônias/metabolismo , Regulação para Cima , Transporte Ativo do Núcleo Celular , Animais , Aquicultura , Autorrenovação Celular , Regulação para Baixo , Receptor alfa de Estrogênio/agonistas , Receptor alfa de Estrogênio/metabolismo , Proteínas de Peixes/agonistas , Proteínas de Peixes/antagonistas & inibidores , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Hidroxiprogesteronas/metabolismo , Células Intersticiais do Testículo/citologia , Células Intersticiais do Testículo/metabolismo , Masculino , Receptores de Progesterona/antagonistas & inibidores , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismo , Células de Sertoli/citologia , Células de Sertoli/metabolismo , Espermatogônias/citologia , Testosterona/análogos & derivados , Testosterona/metabolismo , Técnicas de Cultura de Tecidos/veterinária
12.
Mitochondrial DNA A DNA Mapp Seq Anal ; 28(6): 920-926, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-27608325

RESUMO

The giant river catfish Sperata seenghala has huge demand in South Asian countries due to its low number of intramuscular bones and nutritive value. However, the culture practises for this fish have not been standardized and the current demand for this fish is being met by capture fisheries only. Unregulated and indiscriminate fishing would lead overexploitation of fish stocks subsequently stock depletion. Genetic diversity between populations would give insight about population structure and demography. In the present study, S. seenghala stocks from three rivers, namely Ganga, Brahmaputra and Mahanadi were characterized using cytochrome b gene and D-loop region. Moderate to high haplotype diversity and low nucleotide diversity values were observed in all populations. Analysis of molecular variance and pairwise FST values showed significant genetic differentiation among populations. Patterns of diversity, haplotype networks and mismatch distribution strongly suggest a historical influence on the genetic structure of S. seenghala populations. S. seenghala stocks from these three rivers are genetically distinct units and management measures should be formulated separately for each population.


Assuntos
Peixes-Gato/genética , Variação Genética , Genética Populacional , Genoma Mitocondrial/genética , Animais , Haplótipos , Índia , Análise de Sequência de DNA
13.
Food Sci Technol Int ; 22(3): 266-74, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26199223

RESUMO

The gel properties of sutchi catfish surimi prepared by conventional washing and alkaline-saline washing method were studied for four washing cycles. Decrease (p < 0.05) in myoglobin content was found in alkaline washing process compared to conventional washing at each washing cycle. The highest hardness, breaking force and deformation was observed in gels prepared from alkaline-saline washing method. Whiteness in conventional washed surimi gels increased non-significantly (p < 0.05) compared to alkaline-saline-washed surimi gels. Protein bands on Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that the protein extractability was more in alkaline-saline washing and disappearance of bands in conventional washing method was observed between 116 and 45 kDa indicating less yield of protein.


Assuntos
Manipulação de Alimentos , Géis , Carne/análise , Animais , Peixes-Gato , Géis/química , Concentração de Íons de Hidrogênio , Água
14.
Mol Biol Rep ; 41(5): 2979-87, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24449367

RESUMO

Genes coding for type-I interferon (I-IFN) has been cloned from Labeo rohita, a commercially important and widely cultured fish in India and South East Asia. In the present study, full-length gene of I-IFN was amplified and sequenced. The sequence analysis revealed that I-IFN consists of 1,786 bp genomic sequence with four introns and five exons and an ORF of 546 bp encoding for a putative protein of 181 amino acids. The mature protein has a molecular weight of 18.97 kDa and consists of 158 amino acids and a signal peptide of 23 amino acids at the N terminus. The sequence carries I-IFN signature motif, one glycosylation site, two conserved cystine amino acids and other conserved amino acids. The sequence showed highest similarity to that of Cyprinus carpio (84%). In silico analysis of the rohu I-IFN protein was done using various bioinformatic tools. The constitutive expression of I-IFN gene was found to be more in spleen compared to gill and kidney in real time PCR assay. Expression of I-IFN increased about 20-fold in cultured kidney cell 2 h after induction with poly I:C and showed maximum expression at 8 h post-induction.


Assuntos
Peixes/genética , Expressão Gênica , Interferon Tipo I/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Modelos Moleculares , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Conformação Proteica , Alinhamento de Sequência , Análise de Sequência de DNA
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