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Genet Mol Res ; 13(4): 8982-96, 2014 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-25366789

RESUMO

Biofilm formation of pathogen bacterium is currently one of the most widely studied topics; however, little is known regarding pathogen bacteria biofilms in aquaculture. Aeromonas hydrophila is a representative species of the genus Aeromonas, which has been recognized as a common pathogen, is associated with many diseases in aquatic animals, and causes significant mortality. The objectives of this study are i) to confirm that A. hydrophila can form biofilms on abiotic substrates and construct a biofilm growth curve for this bacterium; ii) to identify the genes that play crucial roles in A. hydrophila biofilm formation. The biofilm growth curve of A. hydrophila was constructed using a crystal violet assay, which showed that biofilm formation for this bacterium is a dynamic process. Next, a mutant library of pathogenic A. hydrophila B11 was constructed using the mini-Tn10 transposon mutagenesis system. A total of 861 mutants were screened, and 5 mutants were stably deficient in biofilm formation. Molecular analysis of the mutant B112 revealed that the open reading frame that encodes the protein MshQ was disrupted. Comparison of biological characteristics including growth, motility, and adhesion between the mutant B112 and the wild-type strain B11 suggested that MshQ is necessary for mannose-sensitive hemagglutinin pilus biosynthesis of A. hydrophila, and that these pili play crucial roles in A.hydrophila adherence to a solid surface during the early stages of biofilm formation.


Assuntos
Aeromonas hydrophila/fisiologia , Proteínas de Bactérias/fisiologia , Biofilmes/crescimento & desenvolvimento , Fímbrias Bacterianas/fisiologia , Aeromonas hydrophila/genética , Aeromonas hydrophila/metabolismo , Sequência de Aminoácidos , Aderência Bacteriana/genética , Aderência Bacteriana/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Elementos de DNA Transponíveis/genética , Fímbrias Bacterianas/genética , Fímbrias Bacterianas/metabolismo , Microscopia Eletrônica de Transmissão , Dados de Sequência Molecular , Mutagênese Insercional , Mutação , Fases de Leitura Aberta/genética , Homologia de Sequência de Aminoácidos , Fatores de Tempo
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