Application of a modified membrane-trapping technique to determine fish microflora species from Thai silver BARB (Barbonymus gonionotus)
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Abstract
A combined method using both culture-dependent methods (CDM) and culture-independent methods (CIM) was used to investigate the fish microflora of Thai silver BARB fish. The species among microflora communities from different parts of fresh healthy fish, gills, flesh, and digestive tracts as well as fish pond water were investigated. Using a traditional CDM, a total of 316 isolates from the cultivation method were randomly selected based on their morphological, physiological, and biochemical characteristics. Most of the acid-producing Gram-positive fermentative bacteria were identified as genera of lactic acid bacteria, Enterococcus, Pediococcus, Streptococcus, Lactococcus, Aerococcus, Tetragenococcus, and Thermobacterium. Moreover, genera of Gram-negative and other Gram-positive bacteria were identified as Enterobacter and Staphylococcus. In the case of CIM, 18 bands appeared using polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE). They were identified based on 16S recombinant DNA (rDNA) sequencing analysis as Weissella confusa, Streptococcus lutetiensis, Enterococcus faecium, Enterococcus gallinarum, Lactobacillus fermentum, Pediococcus acidilactici, and Enterococcus casselifavus. Other Gram-positive enteric bacteria species and some yet uncultured bacteria were also identified as Enterobacter cloacae, Enterobacter ludwigii, Enterobacter mori, Enterobacter roggenkampii, Staphylococcus sp., Staphylococcus epidermidis, Bacillus cereus, Oceanobacillus profundus, and an uncultured bacterium. These results indicate that Enterococcus, Lactobacillus, Streptococcus, and Pediococcus are the predominant species of Thai silver BARB microflora.
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