Environmental air sampling holds significant potential as a tool for viral surveillance. Its use in agricultural and indoor settings has demonstrated its feasibility and effectiveness but despite this, it has rarely been used in wildlife settings. To enable future applications, we optimized key parameters in air sampling methodology using a cave-roosting bat assemblage as a model system. We systematically investigated the impact of sampling conditions (flow rate, sampling duration, and sampling location/deployment time) and post-sampling treatments (DNA/RNA Shield ratios and secondary filtration) on three viral metrics - total mammalian virus abundance, mammalian RNA virus abundance, and Shannon diversity index - generated from next-generation sequencing data. We first showed that air sampling can recover broad viral diversity, including alphacoronaviruses and betacoronaviruses. The sampling conditions for maximizing viral metrics were larger air sample volumes (≥24,000 liters) and sampling inside the cave while the bats were roosting, as opposed to at the cave entrance during emergence. Post-sampling treatments had limited impact on viral metrics, but their application may vary depending on the objectives of the study. This work provides a proof-of-concept for applying air sampling for wildlife viral surveillance in a cave-roosting bat assemblage and identifies key sampling parameters.

Barriers to dog vaccination in Tanzania in general, and in Kilosa District in particular, are not well understood. Therefore, this study sought to address the following research question: What are the multilevel barriers to the vaccination of dogs against rabies in Kilosa District? Guided by a socioecological model, the study explored the barriers to dog vaccination at multiple levels within the Kilosa District context. A case study design employing a qualitative research approach was used. Data were collected through focus group discussions (FGDs) and in-depth interviews to investigate contextual barriers to dog vaccination. The collected data were analyzed using a thematic analysis approach. The results are organized according to the levels of the socioecological model. At the individual level, barriers include limited literacy about rabies and dog rabies vaccination; low perceived risk of rabies transmission; low prioritization of dog vaccination in household financial decisions; and uncertainty regarding the cost of dog vaccination. At the organizational/health system level, barriers include fragmented provision of rabies-related health education, reactive rather than proactive sensitization efforts, and limited communication channels, primarily relying on loudspeaker announcements. At the community level, barriers include misconceptions about protection against rabies and mistrust in political leadership. In conclusion, Rabies vaccination uptake in Kilosa District is constrained by multilevel barriers. Individually, limited knowledge, low risk perception, and financial uncertainty reduce prioritization of dog vaccination. Organizational barriers include fragmented health education, weak cross-sector coordination, and reactive communication. At the community level, logistical challenges, absence of By-Laws, persistent misconceptions, and political mistrust further limit vaccination coverage.

Scrub Typhus accounts for approximately one-fourth of all acute undifferentiated febrile illness cases in India. In September 2023, Odisha, an eastern Indian state, reported several scrub typhus deaths. We investigated a scrub typhus cluster in Block-A, Odisha, to describe the epidemiology and assess risk factors. We conducted an unmatched (1:1) case-control investigation. Line-list of scrub typhus IgM-ELISA tested individuals was collected from the district laboratory. Cases were defined as scrub typhus IgM-ELISA positive residents of Block-A between July1 and October30, 2023 and controls were test-negative block-A residents. We interviewed cases and controls from list of tested individuals for socio-demographic, clinical and exposure histories. We present adjusted-odds ratio and 95% confidence intervals. We conducted an entomological survey in Block-A, isolated mites to detect pathogen and strain (47kDa Real Time PCR) and report entomological indices. Total 151 cases were reported in 2023 in Block-A, with case-fatality rate of 2.6%, median age 30 years, and inter-quartile range 15–46 years. Among the 61 interviewed cases, 46% were hospitalised, 13% admitted in ICU, 16% visited health-facility in second week of symptom onset and 30% were tested in second week of symptom-onset. We enrolled 61 cases and 61 controls, among whom recent farm/forest visit was significantly associated [adjusted OR = 6.1, 95%CI: 2.0-18.4] with illness on multivariate analysis. Chigger-index for block-A was 18.94(322/17); 36%(4/11) of tested mite-pools were positive for Orientia tsutsugamushi and sequences were identified to be Karp-like. Our investigation confirmed a lab-verified scrub typhus cluster in Block-A, identifying farm/forest visits without protective clothing as the most significant factor for acquiring infection. Detection of infected vector, hosts, and pathogen completed the epidemiological triad of scrub typhus, establishing local transmission in Block-A. We sensitised frontline-healthcare workers on early identification and timely referral, recommended community education on limiting mite exposure and seeking timely medical attention. • We aimed to describe the epidemiology of scrub typhus and identify associated risk factors. • We identified recent farm and forest visits as a significant risk factor for scrub typhus infection, and detected infected vectors, maintenance and incidental hosts, along with the pathogen, therefore completing the epidemiological triad and confirming local transmission in this northwestern region of Odisha. • Scrub typhus, a neglected and re-emerging zoonosis affecting large parts of India, including northwestern Odisha, has limited epidemiological evidence in literature; therefore, our one-health investigation, providing comprehensive evidence of local transmission and presence of significant risk factors, addresses this critical gap in the literature, emphasising the urgent need for targeted interventions in these endemic areas.

From July 2024–February 2025, three-quarters of the districts in Uganda experienced mpox outbreaks, with sustained transmission in urban cities and towns. Following the rise in cases and severe outcomes, there was an urgent need to assess sub-national surveillance capacities to inform improvements in mpox response. This study aimed to assess the strengths, weaknesses, opportunities, and threats to mpox surveillance in Uganda’s high-burden districts and document lessons learned for responding to future outbreaks. We conducted a cross-sectional qualitative study during February–March 2025 in 30 purposively selected districts across five high-burden regions of Uganda. Participants were drawn from regional, district, facility, and community levels, including surveillance officers, clinicians, laboratory personnel, and community health leaders. Data were collected through focus group discussions using semi-structured interview guides. Notes were manually coded by two researchers and analyzed thematically using a SWOT (Strengths, Weaknesses, Opportunities, and Threats) framework. Sub-themes were developed inductively, and written informed consent was obtained from all participants. A total of twenty focus group discussions were conducted across five high-burden regions. Participants identified key strengths in mpox surveillance, including functional regional emergency coordination centers, onsite capacity building in digital tools like Go.Data, dedicated surveillance personnel, prior outbreak response experience, and the presence of trusted community health structures. However, notable weaknesses included inaccurate contact information, delayed laboratory test results, workforce shortages, misdiagnosis at lower health facilities, limited digital capacity, inadequate isolation facilities, and stockouts of essential supplies, food, and personal protective equipment. Key opportunities included scaling capacity-building efforts and enhancing surveillance infrastructure, such as call centers and toll-free lines, while surveillance threats included community fear of isolation and strain of concurrent outbreaks such as Ebola. Uganda’s mpox surveillance revealed key strengths, including functional regional coordination hubs, trusted community health structures, and dedicated surveillance personnel that supported early detection. However, gaps such as limited human resources, delayed laboratory results, stockouts, and fear of isolation hindered effective response. Addressing these challenges through investments in decentralized coordination, digital tools, surge staffing, and supply chain systems could enhance future surveillance efforts.

Zoonotic diseases pose an increasing threat to human, animal and environmental health, and identifying the zoonoses of greatest public health concern is key to address them effectively. Despite high ongoing risks from zoonotic diseases, Bhutan has never conducted a zoonotic disease prioritization exercise. Therefore, the present study aimed to prioritize the top 10 zoonotic diseases in Bhutan and develop action plans to support targeted resource allocation, coordinated surveillance, and strategic planning through the One Health framework. The current study used a literature review of peer-reviewed publications and grey literature, expert knowledge, followed by a workshop to prioritize zoonoses in Bhutan. The workshop was convened in April 2024, and 42 participants from 20 different organizations from human, animal, wildlife, environmental, and food safety sectors participated in the workshop. Zoonotic disease prioritization was conducted using a mixed-method approach that combined an expert consensus approach with the U.S. Centers for Disease Control and Prevention One Health Zoonotic Disease Prioritization (OHZDP) tool. The initial literature review listed 55 zoonotic diseases for prioritization in the country. The workshop prioritized the top 10 zoonotic diseases viz., Rabies; Influenza A H5N1; Leptospirosis; Brucellosis; Escherichia coli infections; Dengue; Scrub Typhus; Bovine tuberculosis; Anthrax; and Salmonellosis. Joint action areas for the top five zoonotic diseases were developed to effectively address each disease through the One Health approach in the country. The prioritized zoonoses in Bhutan should be used for strengthening One Health coordination. Key recommendations from the OHZDP workshop emphasized strengthening disease surveillance, building laboratory diagnostic capacity, enhancing intersectoral collaboration, and developing a harmonized, interoperable system for information sharing and reporting. These measures aim to improve real-time data exchange, facilitate joint outbreak investigations, and support evidence-based decision-making for the prevention and control of zoonotic diseases.

Q fever is a globally distributed zoonosis caused by Coxiella burnetii, which is mainly harbored by farm ruminants. Although some studies have reported the presence of C. burnetii in cats, cat-associated Q fever is very rare. In this study, we report the case of a 30-year-old man who was diagnosed with Q fever in May 2024. The diagnosis was based on the results of next-generation sequencing and confirmed by nested PCR. Epidemiological investigations suggest that he had not come into contact with any infection sources other than two cats, one of which was a parturient cat. The blood samples of the cats were collected and tested by nested PCR and immunofluorescence assay. The results showed that the serum of the parturient cat was positive for both phase I and phase II Q fever antigens, suggesting that it had been infected with C. burnetii. This is a rare human case of Q fever transmitted by a domestic cat in China. Importantly, it shows that the pet cat may be a neglected risk factor of Q fever in China.

The Usutu virus (USUV; Orthoflavivirus, Flaviviridae) is a mosquito-borne pathogen causing fatal neuroinfections in susceptible wild and captive birds, particularly blackbirds, other passerines, and owls. Zoological gardens provide favourable conditions for the circulation of such viruses due to the proximity of diverse species and limited options for prevention. Following the sudden death of several Chilean tinamous kept in the Brno zoological garden, we tested tissues sampled from 22 bird cadavers (from zoos, private owners, and free-living birds) for the presence of USUV and West Nile virus (WNV) RNA using duplex reverse transcription qPCR. Near-complete whole-genome sequences were acquired from positive samples by next-generation sequencing and subjected to phylogenetic analyses. Furthermore, serum samples from additional zoo animals and privately owned birds were screened for anti-flavivirus antibodies using ELISA and subsequently confirmed by the virus neutralization test. We report fatal USUV infections in multiple bird species from three zoological gardens in the Czech Republic. Duplex RT-qPCR targeting USUV and West Nile virus (WNV) detected USUV RNA in tissues from two Boreal owls (Aegolius funereus), one Eurasian pygmy owl (Glaucidium passerinum), two Brahminy starlings (Sturnia pagodarum), and four Chilean tinamous (Nothoprocta perdicaria). Additionally, three randomly found cadavers of free-living blackbirds (Turdus merula) tested positive. Pathological findings ranged from minimal pathological changes to pronounced hepatosplenomegaly with intestinal bleeding. Phylogenetic analysis of near-complete genome sequences assigned all viruses to the Europe 2 genetic lineage, revealing partial geographic clustering among isolates obtained in this study. Serological testing confirmed exposure in additional birds and demonstrated cross-neutralisation between anti-USUV and anti-WNV-positive sera. In zoological gardens, flavivirus infections can cause substantial losses, even among rarely bred or endangered species. Given the zoonotic potential of both USUV and WNV, documenting their occurrence in avian hosts is important not only for animal health but also for human disease surveillance from the One Health perspective.

Neurocysticercosis (NCC), caused by the larval stage of Taenia solium, is a leading preventable cause of epilepsy in children, particularly in Sub-Saharan Africa (SSA), where inadequate sanitation practices and limited veterinary control strategies persist. A missing link in the One Health response to NCC has been the absence of communication between neurospecialists and the other partners in control strategies. This narrative review explores how integrating neurological expertise into One Health strategies can enhance the prevention and control of NCC-associated epilepsy in children across the region. We reviewed literature from PubMed/MEDLINE, Scopus, Google Scholar, Embase, Web of Science, Global Health, CINAHL, PsycINFO, and the African Index Medicus, including peer-reviewed articles and organizational reports published between 2013 and March 2025. Out of 1,509 records screened, 28 studies met the inclusion criteria, focusing on neuroparasitosis-associated epilepsy in children across SSA. These included research on disease burden and care challenges (n = 10), pathogenesis (n = 8), One Health control strategies (n = 4), and implementation barriers and solutions (n = 6). NCC remains a major contributor to pediatric epilepsy and associated disability in SSA. A One Health approach informed by direct input from neuro-specialists and better recording of infectious causes of epilepsy can assist teams to implement key strategies, including community education, improved sanitation, food safety measures, pig vaccination, and mass drug administration. Strengthening intersectoral collaboration and healthcare access is critical to reducing NCC burden and improving neurological outcomes for affected children.

Background“One Health,” a concept that highlights the need to bring on board multiple players and actors together to address major health problems, has been proposed to be effective in data gathering to mitigate the menace of antimicrobial drug resistance (AMR). Genomic data on extended-spectrum beta-lactamase-producing Escherichia coli (ESBL-EC) across humans, animals, and the environment are limited in low- and middle-income countries (LMICs), including Ghana.ObjectiveThis study determined the prevalence and patterns of AMR in E. coli from diverse sources, and characterized AMR genes, sequence types (STs), and plasmid replicon types in ESBL-EC.MethodologyIn a cross-sectional study, we randomly collected 1500 specimens from healthy humans, cattle, pigs, lettuce, spring onions, pork, beef, and soil samples, between January 2022 - April 2023. E. coli was isolated by routine culture and confirmed by MALDI-TOF MS. E. coli isolates were screened for their susceptibility against 13 antimicrobial agents and ESBL-production. ESBL-EC isolates were whole-genome sequenced (WGS), and in silico analysis was used to determine AMR genes, sequence types (STs), and plasmid replicon types.ResultOf the 1500 specimens from diverse sources cultured, 140 (9.3%) were positive for E. coli. No E. coli was isolated from lettuce, spring onions, and pork. Fifty (35.7%) E. coli isolates were resistant to three or more of the antimicrobials tested, and 30 (21.4%) were ESBL-EC. The proportion of ESBL-EC identified in healthy humans were 14 (20%), cattle 9 (22.5%), pigs 3 (15%), beef 1 (50%) and soil 3 (37.5%). ESBL-EC isolates were highly resistant to ampicillin (100%), cefuroxime (100%), ciprofloxacin (53.6%), and tetracycline (58.2%). However, all ESBL-EC were susceptible to meropenem. Commonly detected AMR genes were blaTEM-1B (32%), tetA (48%) and sul2 (32%), with majority recovered from healthy human and soil samples. The dominant sequence types found were 12% (3) for ST10, ST 9312, ST 206, and ST 4151. The prevalent plasmid replicon types detected were IncFIB (Apoo1918) (40%) and IncFII (pCoo) (36%).ConclusionWithin the metropolis surveyed, we identified MDR ESBL-EC harbouring various AMR genes and plasmid replicon types with diverse E. coli sequence types in healthy humans, animals, and the environment. The detection of blaCTX-M-15 in agricultural soil isolate is worrisome, emphasizing the need for a “One Health” approach in combating AMR.