Background: Plasmid-mediated resistance to the colistin in poultry is considered as an emerging problem worldwide. While poultry constitutes the major industry in Nepal, there is a paucity of evidence on colistin resistance in Escherichia coli isolates causing natural infections in poultry. This study aimed to explore the prevalence of plasmid-mediated colistin resistance gene, mcr-1 in E. coli isolated from liver samples of dead poultry suspected of E. coli infections. Methods: A total of two hundred and seventy liver samples (227 broilers and 43 layers) from dead poultry suspected of colibacillosis were collected from post-mortem in the Central Veterinary Laboratory (CVL), Kathmandu, between 1 February and 31 July 2019. The specimens were processed to isolate and identify E. coli; an antimicrobial susceptibility test (AST) using disk diffusion method was performed with 12 different antibiotics: Amikacin (30 µg), ampicillin (10 µg), ciprofloxacin (5 µg), chloramphenicol (30 µg), cefoxitin (30 µg), ceftazidime (30 µg), ceftriaxone (30 µg), cotrimoxazole (25 µg), gentamicin (10 µg), imipenem (10 µg), levofloxacin (5 µg) and tetracycline (30 µg). Colistin resistance was determined by agar dilution method and colistin-resistant strains were further screened for plasmid-mediated mcr-1 gene, using conventional polymerase chain reaction (PCR). Results: Out of 270 liver samples, 53.3% (144/270) showed growth of E. coli. The highest number (54%; 109/202) of E. coli isolates was obtained in the liver samples from poultry birds (of both types) aged less than forty days. In AST, 95.1% (137/144) and 82.6% (119/144) of E. coli isolates were resistant against tetracycline and ciprofloxacin, respectively, while 13.2% (19/144) and 25.7% (37/144) isolates were resistant to cefoxitin and imipenem, respectively. In the same assay, 76.4% (110/144) E. coli isolates were multi-drug resistant (MDR). The phenotypic prevalence of colistin resistance was 28.5% (41/144). In the PCR assay, 43.9% (18/41) of colistin-resistant isolates were screened positive for plasmid-mediated mcr-1. Conclusion: The high prevalence of mcr-1 in colistin-resistant E. coli isolates in our study is a cause of concern for the probable coming emergence of colistin resistance in human pathogens, due to horizontal transfer of resistant genes from poultry to human isolates.

The poultry sector contributes four percent to the national GDP of Nepal. However, this sector is under threat with periodic outbreaks of Avian Influenza (AI) subtypes H5 and H9 since 2009. This has been both a public health threat and an economic issue. Since the past few years, outbreaks of AI subtype H9 have caused huge economic losses in major poultry producing areas of Nepal. However, the risk factors associated with these outbreaks have not been assessed. A retrospective case-control study was conducted from April 2018 to May 2019 to understand the risk factors associated with AI subtype H9 outbreaks in Kathmandu valley. Out of 100 farms selected, 50 were "case" farms, confirmed positive to H9 at Central Veterinary Laboratory, Kathmandu, and another 50 farms were "control" farms, matched for farm size and locality within a radius of three km from the case farm. Each farm was visited to collect information using a semi-structured questionnaire. Twelve potential risk factors were included in the questionnaire under the broad categories: birds and farm characteristics, and management and biosecurity status of the farms. Univariable and multivariable logistic regression analysis was conducted and corresponding odds ratios were calculated. Risk factors, associated with AI subtype H9 outbreaks in Kathmandu valley, identified in the final multivariable model were: "farms that have flock size greater than median flock size of study farms (>1500)" (OR = 4.41, 95% CI: 1.53-12.71, p = 0.006), "farms that did not apply rules to wear boots for visitors inside the farms" (OR = 4.32, 95% CI: 1.52-12.29, p = 0.006) and "other commercial farms located within one km periphery" (OR = 10, 95% CI: 1.8-50, p = 0.007). This study showed that outbreaks of AI subtype H9 in Kathmandu valley were associated with a higher population of birds in the farm, poor management practices, and weak biosecurity measures in poultry farms. We suggest improving management practices and increase biosecurity in the farms to reduce incidences of AI subtype H9 outbreaks in Kathmandu valley.

Tuberculosis (TB) in humans is primarily caused by Mycobacterium tuberculosis (M. tuberculosis), with millions of infections and hundreds of thousands of deaths worldwide. It creates a substantial economic burden on the community. Unlike M. tuberculosis, Mycobacterium bovis infects cattle and causes bovine TB, also known as zoonotic TB. People can contract zoonotic TB after consumption of unpasteurized dairy products, handling the sick animals, and via occupational exposures. The association between the zoonotic TB in humans and cattle is not well known in Nepal. The study examined the associated risk factors, including exposure to infected cattle, that contribute to TB's development in human beings in Nepal. The study consists of human and animal subjects. Firstly, a retrospective matched case-control study was conducted at the National Tuberculosis Center (NTC), Bhaktapur, Nepal. A total of 290 people (equal numbers of TB cases and control subjects) were interviewed to obtain information on socio-demographic, behavioral, and occupational risks, including the history of cattle related exposures. Secondly, a cross-sectional study was performed among the cattle owned by the TB-confirmed patients. Comparative tuberculin skin test, rapid antibody test, and ELISA were used in parallel to detect M. bovis infection in cattle. The risk factors for the development of TB in humans were smokers (OR = 4.6, 95% CI: 2.1–10.0, p < 0.001), previous history of TB (OR = 7.9, 95% CI: 3.0–20.6, p < 0.001) and history of cattle exposures (OR = 3.9, 95% CI: 2.1–7.4, p = 0.001). Out of 123 cattle sampled, 12 cattle (9.76%, 95% CI: 5.37–16.76, p < 0.0001) were positive by the tuberculin test, 46 (37.4%, 95% CI: 28.97–46.62, p = 0.007) were tested positive by the rapid test, and 7 (5.7%, 95% CI: 2.52–11.80, p < 0.0001) by ELISA test. The inter-test agreement between the tuberculin and ELISA was very strong (κ = 0.72, 95% CI: 0.48–0.95, p < 0.01). This study indicates that exposure to infected cattle and socio-demographic risk factors can contribute to the development of TB in human beings.

Despite the rapidly growing poultry industry throughout Nepal, the periodic outbreaks of diseases and infections in poultry birds led to huge production loss. The aim of this study was to identify the top ten poultry diseases in Nepal and an analysis of their seasonal distributions. A cross-sectional study was performed to describe the distributions of major poultry diseases diagnosed from April 2018 to April 2019 at Central Veterinary Laboratory, Nepal. Out of 2358 observations recorded at the CVL registry at that period, only 2271 observations qualified for the final analysis. Among 2271, removing the missing values, only 1915 observations were used to describe bird characteristics such as median age and mean flock sizes. Descriptive analysis and graphical representation was performed in R studio (Version 1.0.143) and MS excel 2010 respectively. The top ten diseases identified with highest to lowest incidence were: colibacillosis 26% (584/2271), mycotoxicosis 13% (301/2271), ascites 10% (232/2271), complicated chronic respiratory disease (cCRD) 9% (196/2271), infectious bursal disease (IBD) 7% (155/2271), Newcastle disease (ND) 7% (148/2271), avian influenza (AI) 3% (76/2271), salmonellosis 2% (40/2271), infectious bronchitis 1% (33/2271), coccidiosis 1% (25/2271) and non-specific diseases accounts for 21% (481/2271). Cases of colibacillosis were predominant all year round. Mycotoxicosis was seen mostly during pre-monsoon and monsoon season. Ascites and IBD were common during spring and winter seasons. cCRD was most common during summer and winter months. AI kept changing the disease outbreak pattern but it was most common during spring and summer. The number of cases of Salmonella, IB and coccidiosis were not sufficient to provide the seasonal trend. Identification of common poultry disease and their seasonal distributions is useful in taking preventive measures such as vaccination and good management practices to minimize their incidence in the future.

The poultry industry has transformed into one of the vibrant economic activities in Nepal, contributing to almost 4% of the national GDP. We conducted a cross-sectional survey among the poultry farmers of Kathmandu valley and Chitwan district to assess their knowledge of antimicrobial resistance (AMR); their practice of antimicrobial usage and the biosecurity status of their poultry farms. The survey was done to understand the level of knowledge gap among the farmers regarding successful poultry farming with strict biosecurity protocol and minimal use of antibiotics. A semi-structured questionnaire was prepared to interview a total of 112 farmers, with 56 respondents from each study sites. Only nineteen percent (11/56) of the farmers from the Kathmandu valley and 16% (9/56) of that of Chitwan district knew about the antimicrobial resistance (AMR). Doxycycline (25.9%, 29/112) was the most used antibiotic followed by Tylosin (21.5%, 24/112), Colistin (18.75%, 21/112), Ciprofloxacin (13.4%, 15/112) and Neomycin (12.5%, 14/112) in both districts. Majority of the surveyed respondents lacked the proper awareness regarding the importance of considerate use of antimicrobial drugs, effectiveness of implementing proper biosecurity protocols in the farms, and the increasing challenge of AMR in the present world scenario. The findings and results of this survey identify the critical gaps in the knowledge of the farmers concerning the day-to-day operations in their poultry business. Immediate strategic actions and road-maps are required to solve the burgeoning problem of AMR considering the imprudent use of antibiotics in the poultry sector. Int. J. Appl. Sci. Biotechnol. Vol 10(1): 50-59.

Background Brucellosis is a zoonotic disease caused by Brucella spp. In Nepal, the presence of brucellosis in small ruminants, namely sheep and goats, has impacted farmers’ livelihood and the food safety of consumers. A cross-sectional study was conducted in Rupandehi district of Nepal during January to March 2020 to investigate the seroepidemiology and associated risk factors of brucellosis in the sheep and goat population. Altogether, 19 sheep and 60 goat farms in the district were visited. Owners were interviewed to get information on animals, including their management and movement patterns. Three hundred fifty-seven samples (80 sheep and 277 goat samples) were collected proportionately based on farm sizes. Each serum sample was tested with Rose Bengal Test and ELISA to estimate the seropositivity of brucellosis. Logistic regression was carried out to calculate corresponding odds ratios of each variable associated with detection of brucellosis. Results At the farm level, 31.6% (6/19; 95% CI: 12, 54%) of sheep farms and 3.3% (2/60, 95% CI: 0.9, 11.4%) of goat farms were seropositive to brucellosis. Out of 80 sheep serum samples, 12 (15%; 95% CI: 8.79–24.41%) and out of 277 goat serum samples, three (1.1%; 95% CI: 0.37–3.14%) were seropositive to brucellosis. Age greater than 1.5 years (OR = 5.56, 95% CI: 1.39, 29.38; p = 0.02) and herd size of greater than 100 (OR = 4.74, 95% CI: 1.23, 20.32, p = 0.03) were identified as significant risk factors for seropositivity of brucellosis in the sheep population. While in the goat population, none of the variables was identified as a significant risk factor. Conclusion The study provides evidence that the older sheep and the sheep from the large herds were at higher risk of brucellosis. A control program should be put in place immediately in the sheep population because they may transmit infections to other livestock as they were regularly moved for grazing and selling purposes. Also, strict biosecurity measures should be implemented among pastoralists to prevent brucellosis transmission in them. We suggest further one health-based study to reveal the transmission dynamics of brucellosis between animals and humans.

AbstractPoultry sector contributes to four percent in national GDP of Nepal. However, this sector is under threat with periodic outbreaks of Avian Influenza (AI) subtypes H5 and H9 since 2009. This has been both a both public health threat and an economic issue. Since last three years, outbreaks of AI subtype H9 has caused huge economic losses in major poultry producing areas of Nepal. However, the risk factors associated with these outbreaks have not been assessed. A retrospective case-control study was conducted from April 2018 to May 2019 in Kathmandu Valley to understand the risk factors associated with AI subtype H9 outbreaks. Out of 100 farms selected, 50 were “case” farms, confirmed positive to H9 at Central Veterinary Laboratory, Kathmandu, and other 50 farms were “control” farms, matched for farm size and locality within a radius of three km from the case farm. Each farm was visited to collect information using semi-structured questionnaire. Nineteen potential risk factors were included in the questionnaire under the broad categories: birds and farm characteristics, management aspects and biosecurity status of the farms. Univariable and multivariable logistic regression analysis were conducted to calculate corresponding odds ratios. Identified risk factors associated with AI subtype H9 outbreaks in Kathmandu valley were: “Birds of age 31-40 days” (OR= 11.31, 95% CI: 1.31-98.02, p=0.028), “Older farms operating for >5 years” (OR= 10.9, 95% CI: 1.76-66.93, p=0.01), “Commercial layers farms” (OR=36.0, 95% CI: 0.97-1332.40, p=0.052), “Used stream water to water birds (OR= 5.7, 95% CI: 1.10-30.13, p=0.039)”, “Farms without practice of fumigation after each batch of poultry (OR= 4, 95% CI: 1.44-13.13, p=0.009)., “Farm with previous history of AI (OR= 13.8, 95% CI: 1.34-143.63, p = 0.028), “Did not applied farm boots (OR= 2.58, 95% CI: 0.98-6.80, p= 0.055), “Visitors allowed to enter the farms (OR= 2.5, 95% CI: 1.011-6.17, p = 0.047) and “No foot bath at entry of farms (OR= 3.3, 95% CI: 1.29-8.38, p = 0.013). This study depicts that outbreaks of AI subtype H9 in Kathmandu valley was related to poor management practices and biosecurity in the poultry farms. We suggest improving management practices and increase biosecurity in the farms to reduce incidences of AI subtype H9 outbreaks in Kathmandu valley.

Nepal has an agrarian-based economy where the livestock sector contributes about 12 percent to the national GDP. Rupandehi district is one of the largest dairy pocket areas in the nation, but the disease, such as leptospirosis, posed a negative impact on their production and productivity. Despite the economic importance of cattle leptospirosis, the disease is concerned for the people’s occupational risk and food safety. Although leptospirosis is a priority zoonosis in Nepal, the effective implementation of the control program lacks both in animal and human health. A cross-sectional study was conducted to estimate the prevalence and identify associated risk factors of cattle leptospirosis from March 2019 to April 2020. Altogether, 206 cattle herds located in all the sixteen local levels of the district were visited. The owners were interviewed to record the cattle management system’s information and their knowledge of the zoonotic diseases. A total of 383 cattle serum samples were collected, and an indirect antibody ELISA was performed to estimate the infection status of leptospirosis in the cattle herds. All the data were analyzed by Open epi and R software for the descriptive and analytical study. Bivariate and multivariable logistic regression was applied to assess the potential herd level and animal level risk factors. Out of seven potential herd-level risk factors, the purchased cattle herds (OR: 7.2, 95% CI: 1.24-136.5, p=0.025) and cattle with herd size >10 (OR: 14.92, 95% CI: 2.61-283.38, p=0.025) were identified as significant risk factors for leptospirosis. At the animal level, the cattle taken for grazing in pastureland accessed by the community dog was a significant risk factor (OR: 4.16, 95% CI: 1.13-14.06, p=0.034). Surprisingly, none of the farmers had heard about leptospirosis before. The outcome of this research could be an epidemiological insight in control of priority zoonosis to protect the livestock economy and reduce their negative impact on public health.