oa The Bacteriophage Efficiency and Antibiotics Susceptibility against Escherichia Coli and Staphylococcus Aureus

Introduction

This study was held on the in vitro tests for the bacteriophages and their efficiency comparing with the antibiotics susceptibility in destroying bacteria. Because the development of the resistance to chemotherapeutic agents is becoming an increasing problem. Antimicrobial phage therapy trials have demonstratedphage infection of the increasing incidence resistantbacteria against most or all known antibiotics especially (hospital-acquired) infections.

Rational

This had the potential to facilitate more rational thinking about the phages as antimicrobial therapy for the increasing incidence of bacteria resistant against most antibiotics.

Objectives

Isolating and Identifying the bacteriophage, examining the bacteriophage efficiency against bacteria and comparing them with the antibiotics susceptibility.

Methods

Bacteria isolation and identification: Escherichiacoli and Staphylococcus aureus were isolated from Soba Stabilization Station and subjected to test against bacteriophages isolated from the same location. Susceptibility of isolated bacteria: Toward antibiotics and bacteriophages was determining. One of the virus stocks was Chosen and dispended in the serial dilution the virus sample was mixed with a dense bacterial culture and melted with soft agar and then spread over the surface of a base agar plate and used to infectbacteria. The plaques produced were then counted according to the number adjusted for the dilution to investigate bacteriophage specificity toward thespecific bacteria. Protein profiles of the bacteria and their correspondings phages were done by Sodium dodocyl sulphate polyacrylamid gelelectrophoresis (SDS-PAGE). The Microsoft Excelprogram was used for the statistical analysis, and thebioinformatics programmes UN – SCAN – IT version5 and ImageJ 136b were used for the proteinmolecular mass weight analysis.

Results

Escherichia coli: E. coli showed sensitivity towards: Ciprofloxacin, Pefloxacin, Ofloxacine, Tetracycline, Amikacin, Gentamicin, Piperacillin and Ceftizoxime, the largest inhibition zone was shown with Ciprofloxacin as 29 mm diameter. E. coli wasresistant to Chloramphenicol, Cefotaxime, Co-Trimoxazole and Ampicillin. Staphylococcus aureus: he S. aureus was sensitiveto Lincomycin, Cloxacillin, Ciprofloxacin, Tetracycline, Ofloxacine, Ampicillin/ Sulbactam and Cephalexin and the largest inhibition zone was shown with Lincomycin as 42 mm diameter. S. aureus showed resistant towards: Roxythromycin, Gentamicin, Pefloxacin, Cefotaxime, and Co – Trimoxazole. In broth media the affection of the bacteriophage interactions with bacteria showed increasing of the bacteriophages and decreasing of bacteria due to culture clearance, where the readings of the turbidity for the first and second infection showed statistical significant of E. coli phages samples’ transmission due to place of samples collections; from the anaerobic and facultative ponds P>0.05, facultative and maturation P < 0.05 and anaerobic and maturation P>0.05. Whilst, the S. aureus phages samples’ transmission from the anaerobic and facultative P < 0.05, facultative and maturation P < 0.05 and anaerobic and maturation P>0.05. On solid media the affection of the bacteriophage was recognised by the phage plaque formation on bacterial cultures. The antibiotics susceptibility against the bacteria showed statistical significant P < 0.05 for E. coliand P < 0.05 for S. aureus. The phage proteins were separated by Sodium Dodecyl Poly Acrylamide Gel Electrophoresis technique, the protein profiles of E. coli bacteriophage showed three major bands with molecular weight mass of 47, 34 and 16 kilo Dalton and 34 and 20 kDa for S. aureus phage, the band of 35 kDa was the common shared peak between the phage and the bacterial host due to the bacteriophage lytic cycle. The efficiency of isolated phage against E. coli and S. aureus showed remarkable inhibition of growth of the bacteria at both solid and liquid media this might bedue to physio-chemical changes and difference inmotility of these two bacteria. The mechanical action of bacteriophage on selected bacterial species dependon their receptors that adsorb them to their hosts, there lation between the bacteria and their corresponding phages.

Conclusion

The study showed approximately similar results for the mechanical action of the bacteriophage on the selected bacteria species and the mode of action of antibiotics. It is preferably to manipulate bacterial infections by the Bacteriophage therapy in case of the antibiotic resistant bacteria.