Abstract
Identifying Bacillus cereus with conventional methods is neither specific nor rapid because of the close relatedness of the B. cereus group, hence the need for molecular methods. Genotypic profiling of B. cereus isolates from food was obtained by Random Amplified Polymorphic DNA-polymerase chain reaction (RAPD-PCR) using OPR13 primer. A dendrogram was drawn with the Numerical Taxonomy System of Statistic (NTSYS) software. Thirty of the isolates were subjected to molecular identification by 16S rDNA sequencing. The thirty sequences were deposited in GenBank for accession number. Phylogenetic relationship of the 16S rDNA sequence obtained was carried out with the Multiple Alignment using Fast Fourier Transform (MAFFT) software version 7.0. The evolutionary tree was drawn using the Molecular Evolutionary Genetics Analysis (MEGA 6) software. The dendrogram generated for the RAPD profile showed that all the strains are closely related, with a similarity coefficient of 70%. The isolates were confirmed with 16S rDNA sequencing as B. cereus. The thirty sequences deposited in GenBank were given accession numbers: KX574760–KX574769, KX610811–KX610820, MT757957-MT757963, and MT772282-MT772284. By comparing the phylogenetic relationship, eleven of the strains did not cluster with the reference strains from the GenBank but form distinct clades, which means they are likely to be of different ancestors. Conventional methods rarely differentiate bacteria of the same species into clade, neither can it describe their ancestral lineage. Therefore, it is important to employ molecular methods in identifying bacteria to give detailed information about them.
Highlights
Bacillus cereus is Gram-positive rod, motile with peritrichous flagella, and ubiquitous
Spores are more resistant to dry heat and radiation than vegetative cells, and resistance to heat is more considerable in reduced water activity food [6]. e B. cereus group is made up of seven members [7] in which B. cereus, B. anthracis, and B. thuringiensis are the most significant and are closely related [8]. erefore, identifying members of the B. cereus group with biochemical or physiological characteristics is difficult because these characteristics complicate their accurate peculiarity [9]
Random Amplified Polymorphic DNA (RAPD), which is a form of polymerase chain reaction (PCR), is used to study the genetic diversity of an individual. e short RAPD primers of 10 nucleotides are used to amplify the random sequences of the targeted DNA with low temperature during the International Journal of Microbiology annealing stage
Summary
Genotypic Profiling of Bacillus cereus Recovered from Some Retail Foods in Ogun State, Nigeria, and Their Phylogenetic Relationship. Identifying Bacillus cereus with conventional methods is neither specific nor rapid because of the close relatedness of the B. cereus group, the need for molecular methods. Genotypic profiling of B. cereus isolates from food was obtained by Random Amplified Polymorphic DNA-polymerase chain reaction (RAPD-PCR) using OPR13 primer. Irty of the isolates were subjected to molecular identification by 16S rDNA sequencing. E thirty sequences were deposited in GenBank for accession number. Phylogenetic relationship of the 16S rDNA sequence obtained was carried out with the Multiple Alignment using Fast Fourier Transform (MAFFT) software version 7.0. E isolates were confirmed with 16S rDNA sequencing as B. cereus. E thirty sequences deposited in GenBank were given accession numbers: KX574760–KX574769, KX610811–KX610820, MT757957-MT757963, and MT772282-MT772284. Conventional methods rarely differentiate bacteria of the same species into clade, neither can it describe their ancestral lineage. erefore, it is important to employ molecular methods in identifying bacteria to give detailed information about them
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