RESUMO
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.
Assuntos
Doenças dos Peixes/microbiologia , Infecções por Flavobacteriaceae/veterinária , Flavobacterium/isolamento & purificação , Animais , DNA Bacteriano/genética , DNA Ribossômico/genética , Peixes/classificação , Peixes/microbiologia , Infecções por Flavobacteriaceae/microbiologia , Flavobacterium/classificação , Flavobacterium/genética , Flavobacterium/fisiologia , Água Doce/microbiologia , Índia , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genéticaRESUMO
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.
Assuntos
Animais/classificação , Animais/genética , Animais/isolamento & purificação , Animais/microbiologia , Animais/fisiologia , Animais/veterinária , DNA Bacteriano/classificação , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , DNA Bacteriano/microbiologia , DNA Bacteriano/fisiologia , DNA Bacteriano/veterinária , DNA Ribossômico/classificação , DNA Ribossômico/genética , DNA Ribossômico/isolamento & purificação , DNA Ribossômico/microbiologia , DNA Ribossômico/fisiologia , DNA Ribossômico/veterinária , Doenças dos Peixes/classificação , Doenças dos Peixes/genética , Doenças dos Peixes/isolamento & purificação , Doenças dos Peixes/microbiologia , Doenças dos Peixes/fisiologia , Doenças dos Peixes/veterinária , Peixes/classificação , Peixes/genética , Peixes/isolamento & purificação , Peixes/microbiologia , Peixes/fisiologia , Peixes/veterinária , Infecções por Flavobacteriaceae/classificação , Infecções por Flavobacteriaceae/genética , Infecções por Flavobacteriaceae/isolamento & purificação , Infecções por Flavobacteriaceae/microbiologia , Infecções por Flavobacteriaceae/fisiologia , Infecções por Flavobacteriaceae/veterinária , Flavobacterium/classificação , Flavobacterium/genética , Flavobacterium/isolamento & purificação , Flavobacterium/microbiologia , Flavobacterium/fisiologia , Flavobacterium/veterinária , Água Doce/classificação , Água Doce/genética , Água Doce/isolamento & purificação , Água Doce/microbiologia , Água Doce/fisiologia , Água Doce/veterinária , Índia/classificação , Índia/genética , Índia/isolamento & purificação , Índia/microbiologia , Índia/fisiologia , Índia/veterinária , Dados de Sequência Molecular/classificação , Dados de Sequência Molecular/genética , Dados de Sequência Molecular/isolamento & purificação , Dados de Sequência Molecular/microbiologia , Dados de Sequência Molecular/fisiologia , Dados de Sequência Molecular/veterinária , Filogenia/classificação , Filogenia/genética , Filogenia/isolamento & purificação , Filogenia/microbiologia , Filogenia/fisiologia , Filogenia/veterinária , /classificação , /genética , /isolamento & purificação , /microbiologia , /fisiologia , /veterináriaRESUMO
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.(AU)
Assuntos
Fibra de Lã/classificação , Animais/genética , Animais/isolamento & purificaçãoRESUMO
Two new cell lines (CCF and CCH) were established from fin and heart tissues of common carp, Cyprinus carpio. The cells were optimally maintained in Leibovitz-15 medium supplemented with 10% fetal bovine serum (FBS) and 10 ng/ml of basic fibroblastic growth factor (bFGF). The effects of temperature, concentration of FBS and bFGF on the growth of CCF and CCH cells were examined. The temperature ranged from 24 to 32°C for good growth of the cells. The growth rate of cells was higher in medium containing 10% FBS and the addition of bFGF to the medium significantly increased the growth rate. The CCF cells were found to be epithelial, while the CCH cells were fibroblastic in nature. The cytogenetic analysis of the cell lines revealed a diploid number of 100 chromosomes in C. carpio. The viability of CCF and CCH cell lines were 70 and 72%, respectively, after six months of storage in liquid nitrogen (-196° C). Molecular characterization of the cell lines using 16S rRNA and Cytochrome Oxidase Subunit I (COI) revealed the origin of the cell lines. These new cell lines will be useful for isolation of fish viruses and other in vitro biotechnological studies.
Assuntos
Nadadeiras de Animais/citologia , Carpas , Técnicas de Cultura de Células/métodos , Linhagem Celular/citologia , Miocárdio/citologia , Animais , Carpas/virologia , Bovinos , Sobrevivência Celular , Criopreservação , Cariotipagem , RNA Ribossômico 16S/genética , Temperatura , Fatores de TempoRESUMO
Two new cell lines (CCF and CCH) were established from fin and heart tissues of common carp, Cyprinus carpio. The cells were optimally maintained in Leibovitz-15 medium supplemented with 10 percent fetal bovine serum (FBS) and 10 ng/ml of basic fibroblastic growth factor (bFGF). The effects of temperature, concentration of FBS and bFGF on the growth of CCF and CCH cells were examined. The temperature ranged from 24 to 32 °C for good growth of the cells. The growth rate of cells was higher in medium containing 10 percent FBS and the addition of bFGF to the medium significantly increased the growth rate. The CCF cells were found to be epithelial, while the CCH cells were fibroblastic in nature. The cytogenetic analysis of the cell lines revealed a diploid number of 100 chromosomes in C. carpio. The viability of CCF and CCH cell lines were 70 and 72 percent, respectively, after six months of storage in liquid nitrogen (-196 ° C). Molecular characterization of the cell lines using 16S rRNA and Cytochrome Oxidase Subunit I (COI) revealed the origin of the cell lines. These new cell lines will be useful for isolation of fish viruses and other in vitro biotechnological studies.