THERAPEUTIC EVALUATION OF SYNERGISTIC ACTIVITY OF PROBIOTICS AND Β LACTAM ANTIBIOTICS AGAINST HUMAN PATHOGENS. Dr. Rana Archana S. Assistant Professor Department of Microbiology Shree M. & N. Virani Science college Rajkot (GUJARAT) INDIA

Frame of the Presentation Introduction Antibiotic sensitivity of pathogens Antibiotic sensitivity of probiotics Antibacterial activity of probiotics Synergistic effect of Probiotics and Antibiotics against pathogens Conclusions References Acknowledgement

Introduction

Probiotics World Health Organization: “live microorganisms which when administered in adequate amounts confer a health benefit on the host”

Probiotics Isolates

Antibiotics “Substance produced by one microorganism and inhibitory to other organisms in very minute amount.” Beta-lactam antibiotics

Β Lactam Antibiotics AntibioticsConcentration(ug) cefotaxime/clavu30-10ug aztoeonam30ug cefepime30ug ceftoiaxone30ug cefoxitin30ug entoapenem30ug imipenem30ug cefotaxime30ug ceftazidime/clavu30ug-10ug ceftazidime30ug

ProbioticsAntibiotics Synergistic effect

Human Pathogens Clinical samples- identified by HI-25 Enterobacteriaceae Identification kit Bhatt Laboratory Pathogens Bh-19 Bh-25 Bh-37 Bh-46 Bh-55 Bh-64

Antibiotic sensitivity of pathogens Antibiotic sensitivity of probioticsAntibacterial activity of probiotics Synergistic effect of Probiotics and Antibiotics against pathogens

Antibiotic sensitivity of Pathogens Methodology:  β lactam antibiotics were used to study antibiotic susceptibility.  Disc diffusion method was performed and Zone of inhibition was measured and compared against control.

Antibiotic sensitivity of Pathogens  Graph-1 shows inhibition of the pathogens by different antibiotics.  As compared to others Bh 25 and Bh 64 shows less inhibition.

Antibiotic sensitivity of Probiotic isolates Methodology:  β lactam antibiotics was used to study antibiotic sensitivity.  Disc diffusion method was performed and Zone of inhibition was measured and compared against control.

Antibiotic sensitivity of Probiotic isolates  Graph 2 indicates that vaginal sample isolates were more antibiotic resistant.  Graph 3 shows that few of the baby stool sample isolates were more sensitive as compare to others.  Graph 4,5 and 6 shows that as compare to yoghurt sample and curd sample isolates, milk sample isolates were more sensitive.

Antibacterial Activity of Probiotic isolates Methodology:  Activated cultures of isolates were inoculated in freshly prepared MRS broth and incubated at 37 o C for 24 hrs.  Supernatant was collected by centrifugation at 3000 rpm for 30 min. to study antibacterial activity against selected pathogens.

Antibacterial Activity of Probiotic isolates  Graph 7 indicates that vaginal sample isolates shows more inhibition towards pathogens  Graph 8 shows that few of the baby stool sample isolates shows more inhibition while few not.  Graph 9,10 and 11 shows that yoghurt sample and curd sample isolates, shows more antibacterial activity as compare to milk sample isolates.

Synergism Study Methodology:  Activated cultures of isolates were inoculated in freshly prepared MRS broth and incubated at 37 o C for 24 hrs.  Supernatant was collected by centrifugation at 3000 rpm for 30 min.  Supernatant with Antibiotic used to study synergistic effect against selected pathogens.

Synergism Study  Graph 12,13,14,15 shows Vg5b, Vg5c, Vg5d and Vg6a- vaginal sample isolates synergism with antibiotics to inhibit pathogens  Graph 16 shows Cub inhibition spectrum with combination of β lactam antibiotics.  Graph 17 shows that yoghurt sample isolates in synergism to inhibit pathogens.  All graphs shows that vaginal sample isolates and curd isolates can be the most effective in preparation of synergism with antibiotics.

Conclusions Antibiotics susceptibility of Probiotics it could be concluded that those Probiotics were resistant to all antibiotics, considered as resistant (good). Antibacterial activity against 6 pathogens shows that highly sensitive includes 36.36%, moderate inhibited were 39.39% and very less sensitive or resistant includes 24.24%.

Synergistic effect of antibiotics with Probiotics on pathogens, some of the strain of Probiotics shows increases their effect as compared to single one.

Construction or development of a highly efficient and effective consortium suitable to function under stress and adverse condition can be administered along with commercial one for effective remedy.

Probiotics Antibiotics So from the above discussion it could be concluded that bacteriocin or like other inhibitory compound might be produced by Probiotics has potent action against pathogens. So we can take as a supportive supplement along with antibiotics.

REFERENCES Gerald W. Tannock., A Special Fondness for Lactobacilli, Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand, and Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Canada, minireviews, applied and environmental Microbiology,June 2004, p. 3189–3194 Vol. 70, No. 6. Sami Nikoskelainen,1,2 Seppo Salminen,1 Ran Bylund,2 Ans Arthur C. Ouwehand1., Characterization of the Properties of Human-and Dairy-Derived, Probiotics for Prevention of Infectious Diseases in Fish Department of Biochemistry and Food Chemistry, University of Turku, Turku,1 and Institute of Parasitology, Department of Biology, A°bo Akademi University, Turku,2 Finland Reddy, B. S. and Rivenson, A Inhibitory effect of Bifidobacterium longum on colon, mammary, and liver carcinogenesis induced by 2-amino-3methylimidazo [4,5]quinoline, a food mutagen. Canc. Res. 53: 3914– Gibson, G.R, Dietary Modulation of the human colonic microbiota: introducing the concept of prebiotics. J. Nutr. 125, pp (1995) Huber, J.T pers. comm.

Acknowledgement I am deeply thankful to Dr. Neepa Pandhi for her Guidance. I also acknowledge Management, Principal and staff of Shree M. & N. Virani Science College & Microbiology Department respectively for facilities and support.