A prevalence study was carried to isolate Salmonella typhimurium from blood (n= 50) and gut samples (n=100) of poultry in Kathmandu valley during early 2016. Salmonella typhimurium bacteria isolated in the selective media were biochemically confirmed based on Bergey’s Manual. Two sets of oligonucleotide primers-the genus specific 16S rRNA and the organism specific invA were employed for molecular level confirmation by the Polymerase Chain Reaction (PCR) assay. The amplified fragments in 1% agarose gel observed at 406bp and 285bp, respectively confirmed the isolates to be Salmonella typhimurium. Of 150 samples tested, Salmonella typhimurium were isolated from 49 samples, among which nine were from blood (18%) and forty from the gut (40%). The present result indicated an alarmingly high level of Salmonella typhimurium, which can result inzoonotic infection in humans owing to increased contact with poultry and consumption of poultry products in the Kathmandu valley.

Background Staphylococci are the most commonly isolated pathogen from clinical specimen. These isolates are posing threat due to increasing trend of antimicrobial resistance particularly methicillin. Macrolide-lincosamide streptogramin B family of antibiotics is commonly used to treat such infections. This study was aimed to detect the prevalence of inducible clindamycin resistance and observation of erm and msr genes among Staphylococci isolated from tertiary care hospital of Nepal.Methods Staphylococci from different clinical specimen were identified and antibiotic susceptibility profile were determined following Kirby Bauer disc diffusion method. The double disc diffusion or D-zone test as outlined in CLSI document M100-S24 was performed to examine inducible clindamycin resistance isolates. Multiplex PCR was performed for detection of erm and msr gene in isolates using specific primers for ermA , ermB, ermC, msrA and msrB genes.Results Of the 60 Staphylococci isolates, 39 (65%) were Staphylococcus aureus ( S. aureus ) and 21 (35%) were coagulase negative Staphylococci (CNS) with 25 (64%) and 15 (71%) representing methicillin resistant S. aureus and CNS respectively. Constitutive and inducible MLS B phenotype was observed among 24 (40%) and 14 (23%) isolates respectively by D test. The most prevalent resistant gene was ermC gene (37%) followed by msr B (12%), erm B (10%) and msr A (10%). None of the isolates were found to possess erm A gene.Conclusions The resistant genes were detected more among CNS than S. aureus. The presence of constitutive and inducible MLS B as well as resistant genes among Staphylococci necessitates detection of such isolates to minimize treatment failure. The presence of resistant characteristic varies with hospital settings, geographical locations, patients’ demography etc. The result from this study may help elucidate the predominant resistant characteristics in clinical Staphylococci isolated from tertiary care hospital of Nepal.

BackgroundAntimicrobial resistance (AMR) among bacterial pathogens is a fast-growing public health concern. AMR in non-typhoidal Salmonella species among food animals is of special concern as this may transmit resistant pathogens to humans during handling or consumption of animal products. In Nepal, the possibility of AMR Salmonella species among food animals is an important area of research, particularly in light of the rapidly growing poultry industry, lack of surveillance, and paucity of studies that have been conducted. MethodsTaking one health approach, a cross-sectional study was carried out in Chitwan district of Nepal between May and October 2017. Various environmental samples viz. farm litter, feed, water, poultry feces, vehicle swabs, farm swabs from 12 broiler poultry farms and various sections of poultry carcasses from 21 slaughter houses were aseptically collected. These were microbiologically assessed for the presence of non-typhoidal Salmonella and their phenotypic and genotypic indicators of antimicrobial resistance. ResultsOverall, Of 62 environmental samples collected, 31(50%) tested positive for Salmonella enterica serovars with environmental swabs (70%, 8/12) being the most culture positive sample types. Similarly, of 159 tissue samples collected from 24 carcasses, 79% (126/159) were culture positive for Salmonella enterica. Nearly 97% (153/157), 11% (17/157), 5% (8/157) and none of isolates showed resistance to tetracycline, ciprofloxacin, azithromycin and meropenem respectively. Some 83% (131/157), 40% (64/157), 8% (13/157) and 0.6% (1/157) of isolates tested positive for tetA, QrnS, mefA and VIM-1 AMR genes corresponding to the above antimicrobials respectively.ConclusionsThis study revealed gross contamination of farms and subsequent poultry meat samples with Salmonella enterica serovars that were resistant to several clinically applicable antimicrobials. This reinforces an urgent need to implement proper biosecurity approaches from farms to slaughter houses and strengthen policies to cease the rampant use of clinically important antimicrobials in poultry.

Abstract Background Antimicrobial resistance (AMR) among bacterial pathogens is a fast-growing public health concern. AMR in non-typhoidal Salmonella serovars (NTS) among food animals is of special concern as this may transmit resistant pathogens to humans during handling or consumption of animal products. In Nepal, the possibility of AMR Salmonella serovars among food animals is an important area of research, particularly in light of the rapidly growing poultry industry, lack of surveillance and proper biosecurity measures; and paucity of relevant data. This study was conducted with the aim to estimate the burden of NTS and associated antimicrobial resistance in the environments of commercial poultry farms and the poultry carcasses in slaughter house. This study also intends to find some basic knowledge of the poultry farmers and their practice relating to the use of antimicrobials, vaccination and biosecurity measures.. Methods Taking one health approach, a cross-sectional study was carried out in Chitwan district of Nepal between May and October 2017. Various environmental samples viz. farm litter, feed, water, poultry faeces, vehicle swabs, farm swabs from 12 broiler poultry farms and various sections of poultry carcasses from 21 slaughter houses were aseptically collected. These were microbiologically assessed for the presence of NTS serovars and their phenotypic and genotypic indicators of antimicrobial resistance. The poultry farmers were also briefly interviewed regarding basic their basic biosecurity related knowledge and practices before collecting the environmental samples.Results Overall, of total environmental samples collected, 50% (31/62) tested positive for NTS serovars with environmental swabs (70%, 8/12) being the most culture positive sample types. Similarly, of 159 tissue samples collected from 24 carcasses, 79% (126/159) were culture positive for NTS serovars. Nearly 97% (153/157) of isolates showed antimicrobial resistance to tetracycline, while 11% (17/157) to ciprofloxacin and 5% (8/157) of isolates were resistant against azithromycin. All 157 isolates were sensitive to meropenem. In terms of AMR genes, tetA (83%, 131/157), QrnS (40%,64/157), mefA (8%, 13/157) and VIM-1 (0.6%, 1/157) were detected in the isolates that corresponded to the AMR to tetracycline, ciprofloxacin, azithromycin and meropenem respectively. In farmers interview, only 42% (5/12) of farmers mentioned of using basic biosecurity measures such as applying lime powder around the farm; 84% (10/12) of farmers reported vaccinating their birds with some vaccine and 75% (9/12) of farmers used various antimicrobials prophylactically such as neomycin (33%, 4/12), colistin (33%, 4/12), furaltadone (33%, 4/12), doxycycline (25%, 3/12), sulfatrimethoprim (25%, 3/12) and tylosin (16%, 2/12).Conclusions This study revealed gross contamination of farm environment and subsequent poultry meat samples with NTS serovars that were resistant to several clinically important antimicrobials. Further, inadequacy of even basic biosecurity measures and frequent prophylactic use of antimicrobials in the commercial poultry farms was observed. This reinforces an urgent need to raise awareness and implement proper biosecurity approaches from farms slaughter houses in order to reduce the burden of NTS contamination of surrounding environment and poultry products. Further, high prevalence AMR among NTS isolates also underscores the need to strengthen the policies to prevent the rampant use of clinically used human antimicrobials in poultry sector.