MOLECULAR IDENTIFICATION AND EFFECTS OF CAPSICUM FRUTESCENS EXTRACTS AND ANTIBIOTICS ON BACTERIA IN DOMESTIC FOOD WASTES IN AKURE

Abstract

This project was designed to determine the molecular identification and effects of Capsicum frutescens and antibiotics on bacteria in domestic food wastes (DFW) in Akure. Seventeen bacteria previously isolated from DFW in Akure were provided by the Department of Microbiology, Federal University of Technology, Akure. After purification forty four isolates were obtained. The DNA molecules of the forty four bacterial isolates were extracted using a standard DNA extraction protocol, amplified by Polymerase Chain Reaction and sequenced in a automated ABI Prism DNA sequencer. The bacteria were subjected to antibacterial test using unfiltered extract (UFE) and filtered extract (FE) prepared by grinding and filtering long pepper fruits in sterile distilled water in a ratio 1:2 (w/v) in mortal, pestle and Whatman paper by Agar Well Diffusion Methods. The antibiotics sensitivity profile of the bacteria was determined using standard antibiotics disc employing Disc Diffusion method with Muller Hilton agar. Molecular identification techniques revealed seven Proteus mirabilis, eight Proteus vulgaris, six Baciillus cereus, three Pseudochrobactrum saccharolyticum, one Bacillus subtilis, one Bacillus pumilus, Three Alcaligenes faecalis, one Bacillus anthracis, one Staphylococcus sciuri, one Bacillus thermoleovorans, one Serratia marcescens, three Bacillus licheniformis, three Myroides odoratimimus, one Sphingobacterium mizutaii, one Acinetobacter iwoffii, one Enterobacter cloacae, one Bacillus thuringiensis, one Bacillus weihenstephanensis . Relative to the probable names of the bacteria only two of the bacteria Bacillus subtilis and Bacillus cereus with isolate codes WD3A and WD10A respectively retained their names. The phylogenetic tree showed strong genetic relatedness between Proteus mirabilis and Proteus vulgaris, Sphingobacterium mizutaii and Pseudochrobactrum saccharolyticum. There was genetic dissimilarity between Acinetobacter iwoffii and Myroides odoratimimus, Bacillus cereus and Bacillus pumilus. The zones of inhibition exhibited by UFE and FE were 3.33±0.33 mm to 6.00±0.58 mm and 1.33±0.58 mm to 3.33±0.33 mm. Hence, widest and least inhibitions were observed against Proteus mirabilis, Bacillus licheniformis, Myroids odoratimimus and Proteus vulgaris. Among the commercial Gram positive antibiotics rocephin, erythromycin, ciproflaxin, streptomycin and pefloxacin caused considerable inhibition of the bacteria with zones of inhibition ranging from 3.50±0.50 mm to 12.00±1.00 mm. Zones of inhibition obtained with Gram negative antibiotics were 3.05±0.05 mm to 6.50±0.50 mm with chloramphenicol, gentamycin, sparfloxacin, pefloxacin and ciproflaxacin which showed the widest and smallest zones of inhibition.. This research has revealed correct molecular identities of bacteria associated with DFW in Akure metropolis, Long pepper fruits (Capsicum frutescens var baccatum) inhibitory effect at 0.5 mg/ml and that rocephin, pefloxacin, gentamycin, ciproflaxin and chloramphenicol, erythromycin, streptomycin as the most effective agents against the surviva and growth of these bacteria. Domestic food wastes should be properly disposed off. In order to identify bacteria correctly and to the strain level, molecular technique should be employed since it is the most reliable.