Bioinformatics of blaTEM-1 and blaSHV Genes of Herbal Drugs Pretreated-Klebsiella pneumoniae
Tombari Pius Monsi *
Department of Medical Microbiology, Faculty of Medical Laboratory Science, Rivers State University, Nkpolu-Oroworukwo, Port Harcourt, Rivers State, Nigeria.
Melford Miller Chinwebudu
Department of Medical Technology, College of Allied Medical Sciences, Cebu Doctors’ University, Mandaue City, Cebu, Philippines.
Samuel Douglas Abbey
Department of Medical Microbiology, Faculty of Medical Laboratory Science, Rivers State University, Nkpolu-Oroworukwo, Port Harcourt, Rivers State, Nigeria.
*Author to whom correspondence should be addressed.
Abstract
Background: Oral intake of herbal medicinal products may ultimately lead to their interaction with the intestinal microbial community in which Klebsiella pneumoniae is a resident.
Aim: This study investigated the effect of some herbal medicinal products consumed on beta-lactamase genes, specifically blaSHV and blaTEM, present in a key gut pathogen, K. pneumoniae.
Methods: The study adopted an experimental approach with two strains of K. pneumoniae. Both strains were treated in four herbal medicinal products (Goko bitters, Goko alcoholic bitter, Ruzu bitters and Beta cleanser). The resistant genes, blaSHV and blaTEM, were amplified using polymerase chain reaction on both plasmid and chromosomal DNA. These genes were also sequenced, and presence of mutations were evaluated.
Results: The PCR amplification revealed the presence of chromosomal blaSHV gene in four herbal drug conditions, with bands at 477 bp, including ATCC and clinical strains treated with Goko alcoholic bitters, Ruzu bitters, and control conditions. The blaTEM gene was detected only in the control condition of the clinical strain, marked by a band at 867 bp. Plasmid DNA analysis further confirmed the presence of blaSHV in clinical strains treated with Goko alcoholic bitters and Goko bitters, while blaTEM-1 was observed in strains treated with Goko alcoholic bitters, Goko bitters, and Ruzu bitters. Sequence alignment of the genes blaSHV and blaTEM-1 revealed various nucleotide substitutions, ranging from single nucleotide polymorphisms (SNPs) to length variations when compared to closely related beta-lactamase genes. Mutation analysis indicated that herbal treatments, particularly Goko bitters and Ruzu bitters, induced the highest mutation rates in both blaSHV (11%) and blaTEM (10%), with notable frameshift and point mutations. Conversely, non-herbal medicine-treated conditions displayed fewer or no mutations.
Conclusion: These findings suggest that herbal medicinal products induce stress in K. pneumoniae through promoting the modification of antibiotic-resistant genes. It may also be responsible for the translocation of resistant genes from chromosome to plasmid and vice versa within the bacterial cell.
Keywords: Plasmid, resistant genes, mutation, herbal medicine, bioinformatics