Phylogenetic characterization of canine distemper virus from stray dogs in Kathmandu Valley.

Canine distemper is a highly contagious, often fatal disease caused by canine distemper virus (CDV) in domestic dogs and wild carnivores. The virus has caused mass epidemics in both wild and captive carnivores of high conservation value such as tigers, lions and leopards. Hence, understanding and managing CDV outbreaks is particularly important in Nepal, which is home to many species of threatened wild carnivores including tigers, leopards, snow leopards, dholes and wolves, and also contains a large population of stray dogs. Previous studies have suggested that CDV may pose a threat to wild carnivores, but there have not been any studies characterizing the genetic strains of the virus circulating in Nepal's carnivores. We collected invasive and non-invasive biological samples from stray dogs in Kathmandu Valley and genetically characterized the strains of CDV in the dogs to belong to the Asia-5 lineage by using phylogenetic analysis. The same lineage also contained CDV strains sequenced from dogs, civets, red panda and lions in India. Based on our phylogenetic analysis, we think it is likely that CDV is maintained through sylvatic cycle among sympatric carnivores allowing the recurring spillovers and outbreaks. It is crucial to prevent the virus transmission from reservoir hosts to other species, especially threatened populations of large carnivores in Nepal. Hence, we recommend for regular surveillance of CDV targeting wild carnivores in addition to the domestic dogs.

Assessing antimicrobial resistance in Campylobacter jejuni and Campylobacter coli and its association with antimicrobial use in Canadian turkey flocks.

Turkeys are important sources of antimicrobial-resistant Campylobacter. A total of 1063 isolates were obtained from 293 turkey flocks across Canada between 2016 and 2021 to evaluate their antimicrobial resistance (AMR) prevalence, patterns, distribution, and association with antimicrobial use (AMU). A high proportion of C. jejuni and C. coli isolates were resistant to tetracyclines and fluoroquinolones, despite the very low use of these drugs. C. jejuni isolates had a higher probability of being resistant to tetracyclines than C. coli isolates. The chance of C. jejuni isolates being resistant to fluoroquinolones, macrolides, and lincosamides was lower compared to C. coli. Isolates from the western region had a higher probability of being resistant to fluoroquinolones than isolates from Ontario. Isolates from Ontario had higher odds of being resistant to tetracyclines than isolates from Quebec. No associations were noted between the resistance and use of the same antimicrobial, but the use of certain antimicrobial classes may have played a role in the maintenance of resistance in Campylobacter (fluoroquinolone resistance - bacitracin and streptogramin use, tetracycline resistance - flavophospholipids and streptogramins use, macrolide resistance - flavophospholipid use). Low-level multidrug-resistant Campylobacter was observed indicating a stable AMR in turkeys. This study provided insights aiding future AMU and AMR surveillance.

Characteristics of community-acquired carbapenem-resistant Enterobacterales.

BACKGROUND: Community-acquired carbapenem-resistant Enterobacterales (CA-CRE) are an important threat. METHODS: In CRACKLE-2, we defined patients with CA-CRE as admitted from home, without pre-existing conditions, and a positive culture within 48 h of admission. Healthcare-associated CRE (HA-CRE) were those with the lowest likelihood of community acquisition, not admitted from home and cultured >48 h after admission. Specific genetic markers in carbapenemase-producing Klebsiella pneumoniae were evaluated through random forest modelling. RESULTS: CA-CRE and HA-CRE were detected in 83 (10%) and 208 (26%) of 807 patients. No significant differences were observed in bacterial species or strain type distribution. K. pneumoniae (204/291, 70%) was the most common CRE species, of these 184/204 (90%) were carbapenemase producers (CPKP). The top three genetic markers in random forest models were kpi_SA15, fimE, and kpfC. Of these, kpi_SA15 (which encodes a chaperone/usher system) was positively associated (OR 3.14, 95% CI 1.13-8.87, P = 0.026), and kpfC negatively associated (OR 0.21, 95% CI 0.05-0.72, P = 0.015) with CA-CPKP. CONCLUSIONS: Ten percent of CDC-defined CRE were CA. The true proportion of CA-CRE in hospitalized patients is likely lower as patients may have had unrecorded prior healthcare exposure. The kpi_SA15 operon was associated with the CA phenotype.

MILK Symposium review: Improving control of mastitis in dairy animals in Nepal.

Dairy animals are an important source of income, food, and nutritional security, and improvements in the productivity of dairy animals substantially improve the wellbeing of smallholder dairy farmers. As in other developing countries, dairy animals are key for rural livelihoods in Nepal but often suffer from mastitis-a production disease causing economic losses to farmers, challenges to the dairy processing industry, and possible health hazards to consumers. Studies show that the prevalence of subclinical mastitis in Africa and Asia typically exceeds 50%, threatening animal wellbeing, farmers, dairy processors, and consumers. We conducted a study in Nepal to develop a technology training package to control mastitis in dairy animals. Following identification of knowledge gaps, a technology package consisting of (1) developing good husbandry practices, implementing mastitis detection and control technologies; and (2) training technicians and farmers was implemented. A strategy was subsequently established to provide feedback to farmers in dairy cooperatives on the subclinical mastitis status of their cows. The package was applied in the mid-western region of Nepal. Six months after implementation, we observed a reduction in subclinical mastitis prevalence: from 55% (baseline) to 28% (endline; n = 432) in dairy cows and from 78% to 18% (n = 216) in buffalo. These positive study outcomes strongly suggest that the mastitis technology training package should be scaled across smallholder farmers within and beyond Nepal to control mastitis in dairy animals.

Campylobacter jejuni Strains Associated with Wild Birds and Those Causing Human Disease in Six High-Use Recreational Waterways in New Zealand.

Campylobacter jejuni, a leading cause of gastroenteritis worldwide, has been frequently isolated from recreational rivers and streams in New Zealand, yet the public health significance of this is unknown. This study uses molecular tools to improve our understanding of the epidemiology and sources of Campylobacter in recreational waterways, with a view to preventing human infection. Epidemiological and microbiological data were collected between 2005 and 2009 from six high-use recreational waterways in the Manawatu-Wanganui region of the North Island. Campylobacter spp. and C. jejuni were isolated from 33.2% and 20.4% of 509 samples, respectively. Isolation of Campylobacter was observed in both low and high river flows. After adjusting for the confounding effects of river flow, there was a significantly higher likelihood of isolating Campylobacter in the winter month of June compared to January. A high diversity of C. jejuni multilocus sequence types was seen, with the most commonly isolated being the water rail-associated ST-2381 (19/91 isolates [20.9%]), ST-1225 (8/91 isolates [8.8%]), and ST-45 (6/91 isolates [6.6%]). The ST-2381 was found in all rivers, while the most commonly isolated ST from human cases in New Zealand, the poultry-associated strain ST-474, was isolated only in one river. Although the majority of Campylobacter sequence types identified in river water were strains associated with wild birds that are rarely associated with human disease, poultry and ruminant-associated Campylobacter strains that are found in human infection were also identified and could present a public health risk.IMPORTANCE In 2016, there was a large-scale waterborne outbreak of campylobacteriosis in New Zealand, which was estimated to have affected over 5,000 people. This highlighted the need for a greater understanding of the sources of contamination of both surface and groundwater and risks associated with exposure to both drinking and recreational water. This study reports the prevalence and population structure of Campylobacter jejuni in six recreational waters of the Manawatu-Wanganui region of New Zealand and models the relationship between Campylobacter spp. and ruminant-associated Campylobacter and the parameters "sites," "months," and "river flow." Here, we demonstrate that both low and high river flows, month of the year, and recreational sites could influence the Campylobacter isolation from recreational waters. The presence of genotypes associated with human infection allowed us to describe potential risks associated with recreational waters.

Determination of Serum 25-Hydroxyvitamin D Levels in Patients with Alopecia Areata.

BACKGROUND: Alopecia Areata is a common non scarring alopecia with autoimmune etiology. In several previous studies, an insufficient level of 25 hydroxyvitamin D had been correlated with various autoimmune diseases. The current study aimed to determine the level of serum 25 hydroxyvitamin D in patients with alopecia areata. METHODS: A cross-sectional study was conducted on 35 patients with alopecia areata who visited the outpatient department of Dermatology of KIST Medical College and Teaching Hospital, Nepal. A prior approval of the institute review committee was taken. Convenience sampling technique period for 3 months (March 2022-May 2022) was used to include the participants (informed consent was taken; in minors parental consent was prepared). The serum levels of 25 hydroxyvitamin D was estimated using Abbott Architect i1000 SR analyser. RESULT: In this study, Alopecia Areata was common in the age group of 16-30 years (Mean age: 31.65 ± 11.92 Years). The average duration of the disease in Alopecia Areata patients was close to 1 year. This study indicated a lower concentration of 25 hydroxyvitamin D in half of the studied AA patients (51.4%).The number of patches, Severity of Alopecia Tool scoring, serum 25 hydroxyvitamin D level, and age were found to be statistically insignificant with the disease activity. However, a significant positive correlation was found between the Severity of Alopecia Tool score and the number of patches. CONCLUSIONS: Although low levels of serum 25 hydroxyvitamin D may be seen among subjects with alopecia areata; No statistical correlation was observed. Hence, a larger study of similar nature is needed in the future to elucidate the exact correlation.

Decrease in the prevalence of antimicrobial resistance in Escherichia coli isolates of Canadian turkey flocks driven by the implementation of an antimicrobial stewardship program.

The emergence of antimicrobial-resistant organisms at the human-animal-environment interface has raised global concern prompting governments and various stakeholders to take action. As a part of the stewardship initiative, Canadian turkey producers have implemented an antimicrobial use (AMU) strategy to manage antimicrobial resistance (AMR) in their sector. This study evaluated farm-level AMU and AMR data collected between 2016 and 2021 in major turkey-producing provinces/regions through the Canadian Integrated Program for Antimicrobial Resistance Surveillance to assess the progress of the strategy by characterizing the prevalence of homologous and multidrug resistance (MDR) in Escherichia coli isolated from turkeys. Multivariable mixed-effect logistic regression models assessed temporal and provincial/regional variations in AMR and MDR. Negative binomial regression models examined the temporal and regional variations in the total AMU. The total AMU (measured in mg/kg turkey biomass) significantly decreased in all provinces/regions in 2020 and 2021. Escherichia coli isolates from turkey flocks showed a significant decrease in resistance to gentamicin, sulfisoxazole, and tetracyclines during the six-year study period, consistent with the timing of the AMU reduction strategy. The prevalence of MDR isolates was significantly lower in 2020 and 2021 compared to 2016. Higher prevalence was observed in the Western region compared to Québec and Ontario. Two common AMR patterns were identified: ampicillin-streptomycin-tetracyclines and streptomycin-sulfisoxazole-tetracyclines. These AMR patterns indicate possible cross-resistances (same class), co-selection (unrelated classes) for resistance, or potential carryover of resistance determinants from previous production cycles. The decreasing prevalence of resistance to homologous antimicrobials, MDR, and AMU quantity are suggestive that the turkey sector's AMU strategy is achieving its desired impact. However, antimicrobials previously eliminated for preventive use in turkey flocks and the use of highly important antimicrobials in human medicine suggest that the AMU reduction strategy should be monitored and re-evaluated periodically to mitigate the emergence of MDR bacteria and safeguard animal and public health.

Comparison of antimicrobial resistance among Salmonella enterica serovars isolated from Canadian turkey flocks, 2013 to 2021.

The emergence of antimicrobial resistance (AMR) in Salmonella from turkeys has raised a food safety concern in Canada as certain serovars have been implicated in human salmonellosis outbreaks in recent years. While several studies evaluated AMR in broiler chickens in Canada, there are limited studies that assess AMR in turkey flocks. This study analyzed data collected between 2013 and 2021 by the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) farm turkey surveillance program to determine the prevalence of AMR and differences in resistance patterns among Salmonella serovars recovered from turkey flocks. Salmonella isolates were tested for susceptibility to 14 antimicrobials using a microbroth dilution method. Hierarchical clustering dendrograms were constructed to compare the individual AMR status of Salmonella serovars. Differences in the probability of resistance between Salmonella serovars were determined using generalized estimating equation logistic regression models to account for farm-level clustering. Of the 1,367 Salmonella isolates detected, 55.3% were resistant to at least one antimicrobial and 25.3% were multidrug resistant (MDR) (resistant to ≥3 antimicrobial classes). The Salmonella isolates exhibited high resistance to tetracycline (43.3%), streptomycin (47.2%), and sulfisoxazole (29.1%). The 3 most frequently occurring serovars were S. Uganda (22.9%), S. Hadar (13.5%), and S. Reading (12.0%). Streptomycin-sulfisoxazole-tetracycline (n = 204) was the most frequent MDR pattern identified. Heatmaps showed that S. Reading exhibited coresistance to the quinolone class antimicrobials, ciprofloxacin, and nalidixic acid; S. Heidelberg to gentamicin and sulfisoxazole; and S. Agona to ampicillin and ceftriaxone. Salmonella Hadar isolates had higher odds of resistance to tetracycline (OR: 152.1, 95% CI: 70.6-327.4) while the probability of being resistant to gentamicin and ampicillin was significantly higher in S. Senftenberg than in all the other serovars. Moreover, S. Uganda had the highest odds of being MDR (OR: 4.7, 95% CI: 3.7-6.1). The high resistance observed warrants a reassessment of the drivers for AMR, including AMU strategies and other production factors. Differences in AMR patterns highlight the need to implement serovar-specific mitigation strategies.

Associations between antimicrobial resistance in fecal Escherichia coli isolates and antimicrobial use in Canadian turkey flocks.

Antimicrobial resistance (AMR) in enteric bacteria continues to be detected in turkey flocks and retail products worldwide, including in Canada. However, studies assessing linkages between on-farm antimicrobial use (AMU) and the development of AMR are lacking. This study aims to identify AMU characteristics that impact the development of AMR in the indicator bacteria Escherichia coli in turkey flocks, building on the Canadian Integrated Program for Antimicrobial Resistance Surveillance methodology for farm-level AMU and AMR data integration. Two analytic approaches were used: (1) multivariable mixed-effects logistic regression models examined associations between AMU (any route, route-specific, and route-disease-specific indication) summarized as the number of defined daily doses in animals using Canadian standards ([nDDDvetCA]/1,000 kg-animal-days at risk) and AMR and (2) multivariable mixed-effects Poisson regression models studied the linkages between AMU and the number of classes to which an E. coli isolate was resistant (nCR E. coli ). A total of 1,317 E. coli isolates from a network of 16 veterinarians and 334 turkey producers across the five major turkey-producing provinces in Canada between 2016 and 2019 were used. Analysis indicated that AMR emerged with the use of related antimicrobials (e.g., tetracycline use-tetracycline resistance), however, the use of unrelated antimicrobial classes was also impacting AMR (e.g., aminoglycosides/streptogramins use-tetracycline resistance). As for studying AMU-nCR E. coli linkages, the most robust association was between the parenteral aminoglycosides use and nCR E. coli , though in-feed uses of four unrelated classes (bacitracin, folate pathway inhibitors, streptogramins, and tetracyclines) appear to be important, indicating that ongoing uses of these classes may slow down the succession from multidrug-resistant to a more susceptible E. coli populations. The analysis of AMU (route and disease-specific)-AMR linkages complemented the above findings, suggesting that treatment of certain diseases (enteric, late-stage septicemic conditions, and colibacillosis) are influential in the development of resistance to certain antimicrobial classes. The highest variances were at the flock level indicating that stewardship actions should focus on flock-level infection prevention practices. This study added new insights to our understanding of AMU-AMR linkages in turkeys and is useful in informing AMU stewardship in the turkey sector. Enhanced surveillance using sequencing technologies are warranted to explain molecular-level determinants of AMR.

Socializing One Health: an innovative strategy to investigate social and behavioral risks of emerging viral threats.

In an effort to strengthen global capacity to prevent, detect, and control infectious diseases in animals and people, the United States Agency for International Development's (USAID) Emerging Pandemic Threats (EPT) PREDICT project funded development of regional, national, and local One Health capacities for early disease detection, rapid response, disease control, and risk reduction. From the outset, the EPT approach was inclusive of social science research methods designed to understand the contexts and behaviors of communities living and working at human-animal-environment interfaces considered high-risk for virus emergence. Using qualitative and quantitative approaches, PREDICT behavioral research aimed to identify and assess a range of socio-cultural behaviors that could be influential in zoonotic disease emergence, amplification, and transmission. This broad approach to behavioral risk characterization enabled us to identify and characterize human activities that could be linked to the transmission dynamics of new and emerging viruses. This paper provides a discussion of implementation of a social science approach within a zoonotic surveillance framework. We conducted in-depth ethnographic interviews and focus groups to better understand the individual- and community-level knowledge, attitudes, and practices that potentially put participants at risk for zoonotic disease transmission from the animals they live and work with, across 6 interface domains. When we asked highly-exposed individuals (ie. bushmeat hunters, wildlife or guano farmers) about the risk they perceived in their occupational activities, most did not perceive it to be risky, whether because it was normalized by years (or generations) of doing such an activity, or due to lack of information about potential risks. Integrating the social sciences allows investigations of the specific human activities that are hypothesized to drive disease emergence, amplification, and transmission, in order to better substantiate behavioral disease drivers, along with the social dimensions of infection and transmission dynamics. Understanding these dynamics is critical to achieving health security--the protection from threats to health-- which requires investments in both collective and individual health security. Involving behavioral sciences into zoonotic disease surveillance allowed us to push toward fuller community integration and engagement and toward dialogue and implementation of recommendations for disease prevention and improved health security.

Vulnerabilities for Exposure to Emerging Infectious Disease at Urban Settlements in Nepal.

In Nepal, rapid urbanization and rural-to-urban migration especially due to internal civil conflict have catalyzed the development of temporary settlements, often along rivers on undeveloped land. This study conducted surveillance for viruses in small mammals and assessed potential risks for virus transmission to people in urban settlements along rivers in Kathmandu, Nepal. We collected samples from 411 small mammals (100 rodents and 311 shrews) at four riverside settlement sites and detected six viruses from four virus families including Thottapalayam virus; a strain of murine coronavirus; two new paramyxoviruses; and two new rhabdoviruses. Additionally, we conducted surveys of 264 residents to characterize animal-human contact. Forty-eight percent of individuals reported contact with wildlife, primarily with rodents and shrews (91%). Our findings confirm that rodents and shrews should be considered a health threat for residents of temporary settlements, and that assessment of disease transmission risk coupled with targeted surveillance for emerging pathogens could lead to improved disease control and health security for urban populations. Additionally, interventions focused on disease prevention should consider the unique urban ecology and social dynamics in temporary settlements, along with the importance of community engagement for identifying solutions that address specific multi-dimensional challenges that life on the urban river margins presents.

Local and global genetic diversity of protozoan parasites: Spatial distribution of Cryptosporidium and Giardia genotypes.

Cryptosporidiosis and giardiasis are recognized as significant enteric diseases due to their long-term health effects in humans and their economic impact in agriculture and medical care. Molecular analysis is essential to identify species and genotypes causing these infectious diseases and provides a potential tool for monitoring. This study uses information on species and genetic variants to gain insights into the geographical distribution and spatial patterns of Cryptosporidium and Giardia parasites. Here, we describe the population heterogeneity of genotypic groups within Cryptosporidium and Giardia present in New Zealand using gp60 and gdh markers to compare the observed variation with other countries around the globe. Four species of Cryptosporidium (C. hominis, C. parvum, C. cuniculus and C. erinacei) and one species of Giardia (G. intestinalis) were identified. These species have been reported worldwide and there are not unique Cryptosporidium gp60 subtype families and Giardia gdh assemblages in New Zealand, most likely due to high gene flow of historical and current human activity (travel and trade) and persistence of large host population sizes. The global analysis revealed that genetic variants of these pathogens are widely distributed. However, genetic variation is underestimated by data biases (e.g. neglected submission of sequences to genetic databases) and low sampling. New genotypes are likely to be discovered as sampling efforts increase according to accumulation prediction analyses, especially for C. parvum. Our study highlights the need for greater sampling and archiving of genotypes globally to allow comparative analyses that help understand the population dynamics of these protozoan parasites. Overall our study represents a comprehensive overview for exploring local and global protozoan genotype diversity and advances our understanding of the importance for surveillance and potential risk associated with these infectious diseases.

Infections with multiple Cryptosporidium species and new genetic variants in young dairy calves on a farm located within a drinking water catchment area in New Zealand.

Several Cryptosporidium species are known to infect cattle. However, the occurrence of mixed infections with more than one species and the impact of this phenomenon on animal and human health are poorly understood. Therefore, to detect the presence of mixed Cryptosporidium infections, 15 immunofluorescence-positive specimens obtained from 6-week-old calves' faeces (n=60) on one dairy farm were subjected to PCR-sequencing at multiple loci. DNA sequences of three Cryptosporidium species: C. parvum (15/15), C. bovis (3/15) and C. andersoni (1/15), and two new genetic variants were identified. There was evidence of mixed infections in five specimens. C. parvum, C. bovis and C. andersoni sequences were detected together in one specimen, C. parvum and C. bovis in two specimens, and C. parvum and C. parvum-like variants in the remaining two specimens. Sequencing of gp60 amplicons identified the IIaA19G4R1 (8/15) and IIaA18G3R1 (4/15) C. parvum subgenotypes. This study provides evidence of endemic mixed infections with the three main Cryptosporidium species of cattle and new genetic variants, in calves at the transition age of six weeks. The results add to the body of evidence describing Cryptosporidium isolates as genetically heterogeneous populations, and highlight the need for iterative genotyping to explore their genetic makeup.