Y Zh Xu1, A Anyogu, L I I Ouoba, J P Sutherland. 1. London Metropolitan University, Faculty of Life Science, School of Human Sciences, Microbiology Research Unit, London, UK.
Abstract
AIM: To study genotypic diversity of isolates of Brochothrix thermosphacta recovered from meat, poultry and fish. METHODS AND RESULTS: A total of 27 bacteria isolated from 19 samples of meat, poultry and fish were identified phenotypically and genotypically using PCR amplification of 16S-23S rDNA intergenic transcribed spacer (ITS-PCR), repetitive sequence-based PCR (rep-PCR) and 16S rDNA sequencing. Using ITS-PCR, all bacteria showed the same DNA profile as the reference strains of Br. thermosphacta, allowing typing of the isolates at species level. Using 16S rDNA sequencing, all isolates were identified, at genus and species level, as Br. thermosphacta. Identification as Br. campestris was observed with a lower, but very close, level of similarity. Rep-PCR was more discriminatory than ITS-PCR and allowed differentiation of four subgroups among the isolates. CONCLUSION: Minor genotypic differences among Br. thermosphacta strains from meat, poultry and fish were observed. SIGNIFICANCE AND IMPACT OF THE STUDY: A rudimentary exploration of genotypic differences of Br. thermosphacta from meat, poultry and fish resulted in preliminary confirmation of the suitability of ITS-PCR for typing Br. thermosphacta and confirmed the value of rep-PCR fingerprinting to discriminate between Br. thermosphacta strains.
AIM: To study genotypic diversity of isolates of Brochothrix thermosphacta recovered from meat, poultry and fish. METHODS AND RESULTS: A total of 27 bacteria isolated from 19 samples of meat, poultry and fish were identified phenotypically and genotypically using PCR amplification of 16S-23S rDNA intergenic transcribed spacer (ITS-PCR), repetitive sequence-based PCR (rep-PCR) and 16S rDNA sequencing. Using ITS-PCR, all bacteria showed the same DNA profile as the reference strains of Br. thermosphacta, allowing typing of the isolates at species level. Using 16S rDNA sequencing, all isolates were identified, at genus and species level, as Br. thermosphacta. Identification as Br. campestris was observed with a lower, but very close, level of similarity. Rep-PCR was more discriminatory than ITS-PCR and allowed differentiation of four subgroups among the isolates. CONCLUSION: Minor genotypic differences among Br. thermosphacta strains from meat, poultry and fish were observed. SIGNIFICANCE AND IMPACT OF THE STUDY: A rudimentary exploration of genotypic differences of Br. thermosphacta from meat, poultry and fish resulted in preliminary confirmation of the suitability of ITS-PCR for typing Br. thermosphacta and confirmed the value of rep-PCR fingerprinting to discriminate between Br. thermosphacta strains.