Survey and Phylogenetic Analysis of Rodents and Important Rodent-Borne Zoonotic Pathogens in Gedu, Bhutan.

Rodents are well-known reservoirs and vectors of many emerging and re-emerging infectious diseases, but little is known about their role in zoonotic disease transmission in Bhutan. In this study, a cross-sectional investigation of zoonotic disease pathogens in rodents was performed in Chukha district, Bhutan, where a high incidence of scrub typhus and cases of acute undifferentiated febrile illness had been reported in people during the preceding 4-6 months. Twelve rodents were trapped alive using wire-mesh traps. Following euthanasia, liver and kidney tissues were removed and tested using PCR for Orientia tsutsugamushi and other bacterial and rickettsial pathogens causing bartonellosis, borreliosis, human monocytic ehrlichiosis, human granulocytic anaplasmosis, leptospirosis, and rickettsiosis. A phylogenetic analysis was performed on all rodent species captured and pathogens detected. Four out of the 12 rodents (33.3%) tested positive by PCR for zoonotic pathogens. Anaplasma phagocytophilum, Bartonella grahamii, and B. queenslandensis were identified for the first time in Bhutan. Leptospira interrogans was also detected for the first time from rodents in Bhutan. The findings demonstrate the presence of these zoonotic pathogens in rodents in Bhutan, which may pose a risk of disease transmission to humans.

Determinants of clinical leptospirosis in Nepal.

Leptospirosis is an important zoonotic disease in Nepal; however, there is a lack of information on sources of leptospirosis infection for people and associated risk factors. We implemented a case-control study nested within a cross-sectional survey to investigate zoonotic risks of human leptospirosis among adult, febrile patients seeking treatment in healthcare centres in Kaski District, Nepal, from April to October 2013. The study population was 239 febrile patients; the data consisted of paired blood samples; questionnaires on clinical symptoms; occupational, environmental and animal exposures; and a blood sample from animals in the household. In total, 63 cattle, 92 buffalo, 181 goats, 20 dogs and 14 rodents from 119 households were blood sampled. Serology was performed on the samples using the microscopic agglutination test (MAT) for a panel of 20 serovars with a starting dilution of 1:50. Clinical leptospirosis was defined as a titre of 1:400 or stronger, or a 4-fold or greater rise between acute and convalescent titres, or evidence of seroconversion for one or more serovars represented by a titre of <1:50 in the acute sample and a titre of ≥1:100 in the convalescent sample. The prevalence of clinical leptospirosis was 5.4% (95% CI 2.6%-8.3%). The most common symptoms among recruited participants were loss of appetite and lethargy followed by chills, profuse sweating and backache. No individual clinical symptom nor combination of any two symptoms was significantly associated with clinical leptospirosis. However, owning goats (OR 1.3, CI 95% 1.05-1.66), working in rice fields (OR 1.3, CI 95% 1.11-1.72) and male gender (OR 4, CI 95% 1.12-17.26) significantly increased the risk of clinical leptospirosis in humans. Findings suggest that leptospirosis should be considered in the clinical differential diagnosis among febrile adult patients in Nepal, especially for men, goat owners and/or those who work in rice fields.

One Health research and training and government support for One Health in South Asia.

INTRODUCTION: Considerable advocacy, funding, training, and technical support have been provided to South Asian countries to strengthen One Health (OH) collaborative approaches for controlling diseases with global human pandemic potential since the early 2000s. It is essential that the OH approach continues to be strengthened given South Asia is a hot spot for emerging and endemic zoonotic diseases. The objectives of this article are to describe OH research and training and capacity building activities and the important developments in government support for OH in these countries to identify current achievements and gaps. MATERIALS AND METHODS: A landscape analysis of OH research, training, and government support in South Asia was generated by searching peer-reviewed and grey literature for OH research publications and reports, a questionnaire survey of people potentially engaged in OH research in South Asia and the authors' professional networks. RESULTS: Only a small proportion of zoonotic disease research conducted in South Asia can be described as truly OH, with a significant lack of OH policy-relevant research. A small number of multisectoral OH research and OH capacity building programmes were conducted in the region. The governments of Bangladesh and Bhutan have established operational OH strategies, with variable progress institutionalising OH in other countries. Identified gaps were a lack of useful scientific information and of a collaborative culture for formulating and implementing integrated zoonotic disease control policies and the need for ongoing support for transdisciplinary OH research and policy-relevant capacity building programmes. DISCUSSION: Overall we found a very small number of truly OH research and capacity building programmes in South Asia. Even though significant progress has been made in institutionalising OH in some South Asian countries, further behavioural, attitudinal, and institutional changes are required to strengthen OH research and training and implementation of sustainably effective integrated zoonotic disease control policies.

Comparison of mark-resight methods to estimate abundance and rabies vaccination coverage of free-roaming dogs in two urban areas of south Bhutan.

In Bhutan, Capture-Neuter-Vaccinate-Release (CNVR) programs have been implemented to manage the dog population and control rabies, but no detailed evaluation has been done to assess their coverage and impact. We compared estimates of the dog population using three analytical methods: Lincoln-Petersen index, the Chapman estimate, and the logit-normal mixed effects model, and a varying number of count periods at different times of the day to recommend a protocol for applying the mark-resight framework to estimate free-roaming dog population abundance. We assessed the coverage of the CNVR program by estimating the proportion of dogs that were ear-notched and visually scored the health and skin condition of free-roaming dogs in Gelephu and Phuentsholing towns in south Bhutan, bordering India, in September-October 2012. The estimated free-roaming dog population in Gelephu using the Lincoln-Petersen index and Chapman estimates ranged from 612 to 672 and 614 to 671, respectively, while the logit-normal mixed effects model estimate based on the combined two count events was 641 (95% CI: 603-682). In Phuentsholing the Lincoln-Petersen index and Chapman estimates ranged from 525 to 583 and 524 to 582, respectively, while the logit-normal mixed effects model estimate based on the combined four count events was 555 (95% CI: 526-587). The total number of dogs counted was significantly associated with the time of day (AM versus PM; P=0.007), with a 17% improvement in dog sightings during the morning counting events. We recommend to conduct a morning marking followed by one count event the next morning and estimate population size by applying the Lincoln-Peterson corrected Chapman method or conduct two morning count events and apply the logit-normal mixed model to estimate population size. The estimated proportion of vaccinated free-roaming dogs was 56% (95% CI: 52-61%) and 58% (95% CI: 53-62%) in Gelephu and Phuentsholing, respectively. Given coverage in many neighbourhoods was below the recommended threshold of 70%, we recommend conducting an annual "mass dog vaccination only" campaign in southern Bhutan to create an immune buffer in this high rabies-risk area. The male-to-female dog ratio was 1.34:1 in Gelephu and 1.27:1 in Pheuntsholing. Population size estimates using mark-resight surveys has provided useful baseline data for understanding the population dynamics of dogs at the study sites. Mark-resight surveys provide useful information for designing and managing the logistics of dog vaccination or CNVR programs, assessing vaccination coverage, and for evaluating the impact of neutering programs on the size and structure of dog populations over time.

Building a foundation for 'One Health': an education strategy for enhancing and sustaining national and regional capacity in endemic and emerging zoonotic disease management.

The rapid global spread of diseases such as SARS, H5N1, and H1N1 influenza has emphasized the pressing need for trans-disciplinary collaboration and cross-border action, and has also exposed a serious deficit of capacity and coordination in dealing effectively with emerging disease threats. The need for capacity development is particularly acute in the developing world, which is the least well-equipped to respond adequately. Such capacity development can be achieved through education and the implementation of applied 'One Health' activities. This chapter describes the establishment of a 'One Health' capacity development program in South Asia, consisting of two phases. The first phase provides Masters level training for public health doctors and veterinarians, with a focus on epidemiology, and disease control. The second phase reinforces the postgraduate training by establishing a sustainable framework for the implementation of collaborative 'One Health' activities such as the development of multidisciplinary professional networks, implementation of applied zoonotic disease investigation projects, and support for continuing professional development. The objectives are to provide individual skills required to strengthen capacity; to develop an appreciation of the cross-cutting issues which affect human and animal health, set within an institutional context; and to facilitate the development of regional professional networks which will be instrumental in implementing 'One Health' activities.