Emergence of pathogenic bacteria isolates from zea maize extract using 16s rRNA molecular sequencing protocol as a tools for microbial identification and characterization

Abstract

The purpose of this research work is to determine the molecular identity of bacteria isolated from infected Zea maize using the 16s rRNA molecular sequencing protocol. The samples were obtained from Okeagbe, Akoko north-west local government in Ondo state with latitude and longitude of Okeagbe at 7.6450° N, and 5.7603° E respectively. Preparation of infected maize samples was cultured using the serial dilution method.. Confirmatory characterization of bacteria isolates using 16s rRNA (ribosomal RNA) sequencing procedures (purification, amplication, Sequencing, and DNA extraction) inclusive.The result shows the isolation of the bacteria isolates involved the culturing, inoculation, and plating of the isolate on a plated agar, the identification of the bacteria isolate includes the use of Gram staining, biochemical tests, and characterization using Bergey's manual and antibiotics Susceptibility Test. In Gram staining all bacteria isolates were positive except one, in the biochemical test most bacteria isolate was positive for sugar Fermentation and citrate test and all were negative for the Voges Proskauer test. In antibiotics Susceptibility test few were sensitive, most were susceptible to antibiotics used. With the use of the 16S rRNA and procedures (purification and application of product, Sequencing, and Extraction of DNA) the bacteria isolate were identified and characterized. The phylogenetic analysis and molecular identification of 16S rDNA sequencing revealed that Escherichia coli, Samonella enterica and Staphylococcus aureus were found to infect maize. Molecular characterization based on 16S rRNA Gene sequencing confirms the identity of bacteria. The conventional procedure shows the presence of different arrays of microorganisms in the infected maize, microbes identified are Bacillus subtilis, Bacillus anthracis, Micrococcus luteus, Clostridium sporogenes, Microbacterium lacticum, Clostridium sporogenes, Lactobacillus casei and Micrococcus luteus. The phylogenetic analysis and molecular identification of 16s rRNA sequencing revealed that Escherichia coli, Samonella enterica and Staphylococcus aureus were found to infect maize in Band fragment Base pair 1500bp. In conclusion, the hearsay that maize can only be infected by fungi, it was observed that the possibility of being infected with pathogenic bacteria is imminent. The bottom line is, there should be proper surveillance and food safety in our farm, market and food store, to prevent and totally eradicate emergence of pathogenic organism in our food item.