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Dr. Bhoj R Singh
Indian Veterinary Research Institute, India
Antibiotic and herbal drug resistance patterns of bacteria and emergence of ESBL and NDM strains as cause of the neonatal diarrhea
Neonatal diarrhea is major cause of mortality and economic loss at most livestock farms. For management of diarrhea antibiotic treatment is not a preferred therapeutic line but antibiotics are often a choice if diarrhea lasts for more than three days. However, at the face of emergence of antimicrobial drug resistance in bacteria it is important to look for pattern antibiotic sensitivity and alternate medicine. In the study, 199 bacterial strains (belonging to 21 genera of enteric pathogens) isolated (2012-2015) from referred (having persistent diarrhea) cases of neonatal diarrhea in calves (78), foals (14), buffalo calves (8), goat kids (7), piglets (74), human (7), birds (9) and pups (2) were tested for their sensitivity to 25 antibiotics [to determine multiple drug resistance, extended spectrum B-lactamase (ESBL), carbapenemase and metallo-B-lactamase and New Delhi metallo-B-lactamase (NDM)] and also for sensitivity to 0.5 µL discs of Zanthoxylem rhetsa essential oil (ZREO), thyme essential oil (TEO), ajowan essential oil (AEO), patchouli essential oil (PEO), cinnamon essential oil (CEO), holy basil essential oil (HBEO), carvacrol and cinnamaldehyde (Cin-CHO). The ability to produce ESBL and NDM was also confirmed using specific polymerase chain reaction (PCR) and sequencing of the PCR products. None of the 25 conventional antimicrobials was effective on all the bacterial isolates, the most effective being tigecycline (83.7%), chloramphenicol (81.2%), moxalactam (81.1%), gentamicin (74%), colistin (71.4%), and nitrofurantoin (68.5%), and the least effective ones were erythromycin (1.3%), ampicillin (17.1%), amoxicillin (26.8%) and nalidixic acid (41.3%). More than 60 isolates had resistance to three or more commonly use antibiotics (MDR), while 38.2%, 29.6% and 12.1% strains were positive for ESBL, carbapenemase and NDM production. Among the alternate antimicrobial herbal products tested, PEO was the least effective inhibiting only 6.6% strains followed in increasing antimicrobial activity by ZREO (21.3%), TEO (87.1%), carvacrol (91.9%), HBEO (92.5%), , CEO (96%), cinnamaldehyde (98.2%) and ajowan essential oil (100%). Only 7.5% strains had multiple herbal antimicrobial resistance (MHADR). No significant difference in sensitivity of bacteria associated with diarrhea in different species of hosts to most of the antimicrobials was evident except the higher sensitivity of isolates from foals than those from calves to ampicillin (p, 0.02), tetracycline (p<0.01), cotrimoxazole (p<0.01), ciprofloxacin (p,0.01), amoxicillin (p<0.01), and aztreonam (p, 0.045), while isolates from piglets than of calves were more often sensitive to nalidixic acid (p<0.01) and cefepime (p, 0.01) but trend was opposite for moxalactam (p, 0.02). The ESBL producing strains were more often detected in cases of calf diarrhea (p, 0.03) than those of other animals. The details of trends of emerging antimicrobial drug resistance may be discussed in light of antimicrobial drug uses in animals in the manuscript. Keywords: Enteropathogens, Escherichia coli, Edwardsiella tarda, Aeromonas, Salmonella, MDR, MHADR, NDM, ESBL, Carbapenemase.

He is a microbiologist, graduated in Veterinary Sciences and earned Ph.D. in Microbiology and Public Health is working as molecular epidemiologist having with more than 25 years of teaching and research experience.He has guided nine students for their dissertations for Master in Veterinary Sciences (Microbiology, Bacteriology, Epidemiology) and three students for Ph.D. (Bacteriology), presently three students are working with him, one for MVSc. and two for Ph.D. He published three books on different aspects of Microbiology, about 170 research papers and 9 reviews in journals of international repute. His major contributions in recent past are on culturomes (cultivable microbial diversity) studies in foods and animals of North Eastern India and proved vertical transfer of microbes in house lizards. He also developed disease models for understanding host pathogen interaction including germinating seed model for Salmonellacytotoxins, mouse model for edwardsiellosis and guinea pig model for equine salmonellosis. Besides, he headed the projects for development of genetically defined vaccine for control of salmonellosis in equids, toxoid vaccines for control of salmonellosis and klebsiellosis, genus specific ELISA for diagnosis of salmonellosis and klebsiellosis. He contributed for monitoring and control of veterinary vaccine quality in India. He also established diagnostic facilities for edwardsiellosis, bordetellosis, glanders, and tuberculosis at IVRI, Izatnagar and diagnosis of Canine Parvo virus infections at GB Pant University of Agric. &Technology, Pantnagar.

Dr. Singh’s specific interests are onhost pathogen interactions;usingconventional microbiological, immunological and pathological methods, signature tag mutagenesis, and site directed mutagenesis. He developed several biomodels for studies on animal pathogens. He developed epidemiological tools for molecular typing, pathotypingand toxinotypingof Salmonella. His group is working on host pathogen interaction having focus on detection of pathogens’ signatures in host system including environment using host serum and environmental samples.Besides, targetting research on synergy among antibiotics and herbal antimicrobials to identify newer antimicrobial therapy approaches and molecules. His team explored epidemiology of emergence of antimicrobial drug resistance in bacteria of veterinary clinical significance. He designed Ph.D. courses for Epidemiology, including Molecular Epidemiology as chairman of the faculty of epidemiology. As faculty of Bacteriology,he developed courses on Bacterial Genetics, Antibiotics and Antimicrobial Chemotherapy, Microbiology of Vegetable Foods, Bacterial Toxins, and Enterobacteriaceae for post graduate students at IVRI, Izatnagar.He has good experience of working at BSL-III, ABSL-III and BSL-III+ laboratories as scientist and administrator, at CCS-NIAH, Baghpat (as director), IVRI, Izatnagar and IAH, Compton UK (as scientist).

He is two time recipient of Dr. CM Singh best paper awards, and Award of Honour for research at Indian Veterinary Research Institute, best research article in Hindi (2015, 2016, RajbhashaSmarika, ICAR, New Delhi); consistent reviewer of research proposals for Department of Science and Technology and Department of Biotechnology, New Delhi; for National Center of Science and Technology Evaluation, Ministry of Education and Science, Astana, Republic of Kazakhstan since last several years. He is an International faculty on Enteric Diseases atUniversity of Sassari, Italy.He established microbiology laboratories at National Research Centre on Equines, Bikaner and at ICAR Research Complex for NEH Region, Nagaland, and a Molecular Epidemiology Laboratory at Indian Veterinary Research Institute, Izatnagar. He worked in different capacities starting from field veterinary doctor, Head Division of Animal Sciences, Head Division of Epidemiology, acting Director of CCS National Institute of Animal Health, Baghpat, India. He startedfunctioning at National Institute of Animal Health in 2014as vaccine quality monitoring facility after several years of its non-functional existence.

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