One Health

The emergence of the novel monkeypox virus (MPXV) clade Ib in the Democratic Republic of the Congo (DRC) and neighboring countries in late 2023 highlighted the need for rapid, scalable surveillance approaches to support outbreak detection and response. As part of the ODIN-Mpox project, wastewater surveillance (WWS) systems were established as an emergency public health measure in three Sub-Saharan African countries (DRC, Tanzania, and Burkina Faso) to evaluate the feasibility of wastewater-based monitoring for mpox and strengthen local surveillance capacity. Between January 2025 and April 2026, 117 wastewater samples were collected from selected sites and analyzed for MPXV DNA using targeted qPCR assays. Clinical mpox data were obtained from national surveillance systems and WHO reports to assess epidemiological linkages between wastewater detections and reported infections. Six wastewater samples tested positive for MPXV DNA. During the study period, DRC experienced the highest disease burden, with weekly reported cases peaking at about 3,000 in January 2025, while Tanzania reported a peak of 20 weekly cases in March 2025. No confirmed clinical cases were reported in Burkina Faso. No clear relationship was observed between reported case numbers and qPCR Ct values in positive wastewater samples. Despite the low detection frequency, the project demonstrated the operational feasibility of implementing MPXV wastewater surveillance in resource-limited settings and established laboratory capacity for environmental monitoring of emerging infectious diseases. Given the early stage of WWS implementation in the region, the study identified opportunities for further system strengthening, including optimization of sample processing and reporting workflows, improved access to laboratory supplies, and enhanced integration of environmental and clinical surveillance data streams. These findings highlight the value of WWS as a complementary component of integrated public health surveillance systems and emphasize the need for continued investment in laboratory capacity, harmonized methodologies, governance frameworks, and knowledge exchange to enhance outbreak preparedness and response in low-resource settings.

Mass gatherings pose significant public health risks by facilitating the spread of infectious diseases. While wastewater-based surveillance (WBS) has been widely used to monitor pathogens in high-income settings, its use as a practical, multi-pathogen surveillance tool during mass gatherings in low- and middle-income countries remains limited. This study aimed to assess the operational feasibility, epidemiological significance, and public health utility of multi-pathogen WBS during the African Nations Championship (CHAN) football tournament in Uganda. Wastewater surveillance was conducted at Mandela National Stadium during eight match days in August 2025. Moore swabs were deployed at 38 manholes receiving wastewater from different toilet facilities across the stadium to capture representative wastewater samples. Samples were processed using Nanotrap(R) microbiome virus particles to concentrate pathogens, followed by nucleic acid extraction. Samples were analyzed for multiple enteric and respiratory pathogens, including Mpox, using quantitative PCR (qPCR). Descriptive analyses were performed to characterize pathogen detection patterns, positivity rates, and temporal distribution across surveillance sites. A total of 304 wastewater samples were collected and analyzed, of which 259 (85.2%) tested positive for at least one pathogen. Multiple pathogens were consistently detected across sampling days, with enteric pathogens predominating, particularly Shigella spp. (53.6%), Rotavirus A (35.9%) and Enterovirus (32.2%). The mpox virus was also detected in a notable proportion of samples (28.6%) across several sampling days. Respiratory pathogens, including SARS-CoV-2 (11.8%) and Influenza B (8.2%), were identified intermittently at lower frequencies. Pathogen diversity varied over time, with up to eight pathogens detected on a single day, and co-detection of multiple pathogens observed in the majority of positive samples. Cq value distributions further demonstrated variability in detected signal patterns across pathogens. Surveillance findings informed real-time public health interventions, including sanitation reinforcement, intensified hygiene promotion, environmental disinfection, and targeted risk communication, strengthened syndromic surveillance with on-site triage, and targeted environmental health assessments of food handling and wastewater infrastructure. These findings demonstrate the operational feasibility and public health utility of integrating multi-pathogen wastewater-based surveillance into mass-gathering preparedness and response frameworks in low-resource settings. By capturing diverse pathogen signals and informing targeted interventions during the CHAN football tournament, WBS can provide actionable population-level insights that can support outbreak preparedness and response. Scaling WBS within national preparedness systems could strengthen epidemic intelligence, enhance early warning capacity, and support data-driven public health decision-making during future mass gatherings and emerging infectious disease threats.

The emergence and spread of vector-borne pathogens, such as the mosquito-borne parasite Dirofilaria immitis, the causative agent of canine heartworm disease pose increasing challenges for public and veterinary health. These parasites have expanded from Southern Europe into Central and Eastern Europe, where locally acquired transmission is increasingly documented in pets and wild animals, and is now considered an emerging infectious disease. Understanding the dynamics of its emergence and public awareness is essential for effective surveillance and intervention. Here, we evaluated whether human online search behavior, as measured by Google Trends (GT), reflects patterns of dirofilariosis emergence in Hungary. We analyzed GT data for heartworm-related terms from 2012 to 2025, exploring geographical and temporal trends. Our analyses revealed a sustained increase in search interest over time beginning in 2012, with pronounced rise after 2015. Spatially, the highest relative search volumes were concentrated in southeastern Hungary. We found a strong positive association between regional search activity and mosquito infection rate. These findings demonstrate that GT-derived data accurately mirror both the temporal increase and geographic distribution of canine dirofilariosis, supporting the use of digital epidemiology as a complementary surveillance tool to inform public health and veterinary strategies for emerging vector-borne diseases.

Leptospirosis is a globally important environmentally transmitted disease, with approximately one million cases and 60,000 deaths reported annually. In low-income urban communities, inadequate sanitation, drainage and waste management may increase human exposure to contaminated environments. This study investigated the influence of environmental engineering risk factors on Leptospira exposure in four disadvantaged urban communities (favelas) in Salvador, Brazil. A high-precision georeferenced field survey was developed to identify, map and characterise sanitation, stormwater drainage and solid waste infrastructure. Cross-sectional spatial analyses of baseline data were used to assess associations between environmental risk factors and the residential locations of individuals with anti-Leptospira antibodies. Seropositive individuals tended to reside closer to environmental risk factors and at lower relative elevations. Density analyses indicated that seropositive individuals tended to reside closer to inadequate or partially adequate sewerage components than to sewage-contaminated streams or open sewage points. Inadequate streets showed also showed high density peaks, suggesting that exposure may occur through frequent contact with contaminated runoff and standing water. In contrast, open waste dumping sites and vacant lots showed weaker and more diffuse spatial patterns. The findings highlight the importance of infrastructure quality shaping leptospirosis risk within urban informal settlements. The proposed methodology provides a practical approach for high-resolution characterisation of environmental exposure pathways and may support targeted engineering interventions and epidemiological investigations of leptospirosis and other environmentally transmitted diseases.

Background. Direct livestock exposure is a risk factor for zoonotic influenza, including H5N1 highly pathogenic avian influenza (HPAI) A virus. But whether living in regions of high poultry and swine production intensity (PPI, SPI) increases risk of exposure to zoonotic influenza viruses independent of occupational livestock contact remains unclear. Objectives. To determine whether livestock workers and community members with no occupational livestock exposure in North Carolina, where poultry and swine production are increasingly co-located, are at higher risk of exposure to zoonotic influenza. Methods. Saliva samples from industrial livestock operation worker (ILO-W), ILO neighbor (ILO-N) and metropolitan area (Metro) households were analyzed for mucosal influenza A (H5N1, H1N1, and H3N2) hemagglutinin (HA) IgA and IgG antibodies to determine associations of PPI, SPI, exposure group, and detection of a swine-specific fecal contamination marker (Pig-2-Bac DNA) with influenza A antibody levels. Results. Residing in the highest PPI and SPI tertile was associated with significantly higher mucosal H5 and H1 HA IgA levels, including among residents without occupational livestock exposure. Households with occupational poultry or swine contact had significantly higher H5 IgA and IgG and H1 IgA levels compared to Metro households. In regression models accounting for clustering at the participant level, log10 anti-H5 HA mucosal IgA increased 0.16 (95% CI: 0.06, 0.27, p<0.005) and 0.10 (95% CI: 0.03, 0.17, p<0.005), per log10 increase in PPI and SPI, respectively, and 0.16 (95% CI: 0.03, 0.19, p<0.02) when Pig-2-Bac DNA was detected on household surfaces. Conclusions. Mucosal H5 HA IgA and IgG and H1 HA IgA were consistently elevated across different metrics of livestock exposure intensity, including residential exposure, occupational contact within a household, and a molecular marker of household swine fecal contamination in a state with intensive poultry and swine production.

Antimicrobial resistance (AMR) is one of the greatest challenges within the One Health continuum. Exploring transmission routes between health domains and determining their driving forces are key priorities for future research. This effort can be effectively supported by landscape epidemiology, a field of science that integrates methods from landscape ecology and epidemiology to unravel the complex interdependencies behind disease transmission. This exploratory study aimed to demonstrate that landscape diversity and the degree of hemeroby (anthropogenic impact) correlate with the composition of bacterial communities and their AMR profiles. To test this hypothesis, submandibular lymph nodes from Cervidae and Suidae were collected to detect Staphylococcus and Mammaliicoccus bacteria and characterise their AMR features using selective culture and the VITEK 2 Compact automated system. As a result, the bacterial community in the more natural landscape was more diverse, characterised by the dominance of Mammaliicoccus sciuri and pan-susceptible isolates of Staphylococcus hyicus, and it displayed a low-level, heterogeneous AMR profile. Within the more hemerobic landscape, the bacterial community was characterised by the dominance of Staphylococcus epidermidis, a human-adapted species, and the AMR profile showed signs of higher antimicrobial pressure from both public health and veterinary origins. Although this study was based on only two study sites and was therefore not suitable for drawing definite conclusions, the findings suggest that human impact manifests itself in both bacterial and AMR profiles. A high prevalence of mammaliicocci and a heterogeneous AMR profile appeared to be indicators of naturalness. Conversely, the dominance of a human-adapted bacterial species and the accumulation of AMR features characteristic of medical environments likely indicate higher degrees of hemeroby.

Household environments in low-resource settings can become contaminated with fecal matter from multiple sources, including humans and domestic animals. Identifying the source of fecal contamination is critical for designing targeted interventions to reduce exposure to enteric pathogens, particularly for young children who bear the majority of the enteric disease burden. Using data from 140 households with children enrolled in the ECoMiD cohort study in northwestern coastal Ecuador, we (1) characterized source-specific fecal contamination in samples from household floors and maternal and child hands, and (2) identified animal-related and Water, Sanitation and Hygiene (WASH) conditions associated with the presence and concentrations of these markers. We used five qPCR-based microbial source tracking (MST) markers to detect fecal contamination from avian (GFD), canine (DG37), swine (Pig2Bac), ruminant (Rum2Bac), and human (HF183) sources. Prevalence ratios (PR) and mean differences comparing the presence/absence and concentration, respectively, of MST markers between households with and without each animal-related or WASH condition were estimated using generalized linear models with Poisson and Gaussian distributions. Animal MST markers tracked strongly with several animal-related conditions, whereas associations between the human MST marker and household demographic and WASH conditions were more limited. Animal ownership (PR 1.53; 95% CI: 1.04-2.26) was associated with higher prevalence of animal MST markers on floors. Households reporting animal feces indoors had higher prevalence of animal MST markers on floors (PR 1.84; 95% CI: 1.17-2.89), and higher concentrations of animal MST markers on child hands (mean difference 0.27 gene copies (gc)/m2; 95% CI: 0.12-0.41) and maternal hands (mean difference 0.10 gc/m2; CI: 0.02-0.18). Animal feces left unremoved outside the home were associated with higher prevalence of animal MST markers on maternal hands (PR 2.31; 95% CI: 1.11-4.81). Mothers reporting direct contact with animals had higher concentrations of animal MST markers on their children hands (mean difference 0.12 gc/m2; 95% CI: 0.02-0.21). Higher FECEZ scores, an overall metric for animal exposure, were associated with higher prevalence of animal MST markers on maternal hands (PR 2.94; 95% CI: 1.17-7.40). For human fecal contamination, the presence of E. coli on child hands was associated with higher prevalence of human MST markers on floors (PR 1.30; 95% CI: 1.00-1.68). Our findings reinforce household floors and maternal and child hands as key reservoirs of fecal contamination and point to future potential targets worth exploring for interventions in similar high-burden settings.

Although a strand of scholarship on pandemic prevention flourished in the wake of the COVID-19 pandemic, a theoretically informed empirical analysis of global governance entrepreneurs and practitioner perspectives is lacking. This gap is salient given the need to consider the nuances, political realities, and feasibility of real-world governance practice, particularly with the recent adoption of the Pandemic Agreement under the World Health Organisation. In this paper, nexus governance and regime complex theory guides an analysis of recommendations for potential real-world governance responses for pandemic prevention from wildlife trade for human consumption elicited from global governance entrepreneurs and practitioners through semi-structured interviews and document analysis. Recommendations on future governance practice largely focused on strengthening coordination across various policy sectors to improve use of existing institutional arrangements, with particular emphasis on better integration of the biodiversity conservation policy sector within global pandemic prevention governance, as well as reform of the World Organisation for Animal Health and the Convention on International Trade in Endangered Species of Fauna and Flora. With governance deficits for prevention of pandemics emerging from the wildlife trade left by the now largely concluded Pandemic Agreement, a renewed research agenda on shared governance pathways becomes paramount.

Climate and land use change are reshaping the dynamics of vector-borne diseases. West Nile virus (WNV), the most widespread zoonotic arbovirus in the United States, illustrates the need to integrate climate, land cover, and social vulnerability across heterogenous landscapes when assessing spatial risk. We present a nationwide, county-level assessment of WNV risk, using complementary statistical and mechanistic models to (1) identify socio-ecological correlates of current WNV incidence, and (2) project vector species-specific, temperature-dependent transmission suitability under mid- and late-century climate change scenarios. We find that land cover gradients, temperature-driven transmission, and both occupational and residential exposure are associated with WNV incidence, particularly in mixed urban-agricultural landscapes. Future temperature and land cover projections suggest spatially variable shifts in environmental risk, driven by divergent physiological responses among Culex species vectors. Our results highlight temperature and land cover as consistent, mechanistically grounded correlates of WNV risk at the national scale, while underscoring the need for refined, species-specific analyses at local levels. These insights can inform more targeted surveillance, vector control, and climate adaptation strategies. We also identify key knowledge gaps, particularly around host and vector ecology, that must be addressed to improve public health response in the face of ongoing environmental change.

Introduction: Antimicrobial resistance poses a major global health threat, yet evidence on its economic impact in low- and middle-income countries remains limited. This study estimated the economic burden of infections caused by ESBL-producing Escherichia coli (E. coli), a key resistant pathogen, in Laos. Methods: A prospective cost-of-illness study was conducted among patients with laboratory-confirmed infections in Setthathirath hospital in Vientiane, Laos, collecting cost data through repeated interviews and medical records. Descriptive analyses and econometric modeling approaches, including inverse probability weighting (IPW) and instrumental variable (IV) analyses, were used to estimate out-of-pocket expenditures, public expenditures, total cost of illness, and length of stay, accounting for potential confounding. Results: ESBL-producing E. coli was consistently associated with higher economic burden across all analyses. The unadjusted per-patient cost was US$ 689.0 for ESBL-producing E. coli, compared with US$ 489.4 for non-ESBL-producing E. coli and US$ 537.6 for non-E. coli. The association remained statistically significant for out-of-pocket cost after IPW-adjustment (US$156.2; 95% CI, 14.3 to 298.1; P = 0.03), while other outcomes were not statistically significant. Instrumental variable analyses showed consistent directional effects but with wide confidence intervals and no statistically significant differences. Conclusions: Findings suggest that ESBL-producing E. coli may be associated with increased economic burden in Laos; however, this association was not consistently statistically robust across analytical approaches. These findings suggest a potential economic impact of ESBL infection, although uncertainty remains regarding the magnitude of the effect. Strengthening antimicrobial stewardship, infection prevention and control, and improved diagnostic capacity remain essential to mitigate the potential AMR burden.

Abstract BACKGROUND West Nile virus (WNV) is a growing health burden in Italy. Anticipating human infection risk is hampered by the pathogen's complex ecology, highlighting the need for comprehensive early-warning tools. AIM We aimed to model municipal-level WNV risk in Italy and characterize its decadal expansion in Italy, providing a comprehensive ecological understanding of viral emergence. METHODS We applied a machine learning framework to annual human WNV case data from 2014 to 2024. The model integrated a suite of environmental, socio-economic, and macroecological predictors to generate risk projections. We evaluated the model's performance through multiple validation settings. We also performed an anticipation test for the 2025 epidemic season, using 2024 environmental data to assess the model's predictive accuracy against observed 2025 human cases. RESULTS Our model achieved robust performance (True Skill Statistic > 0.4) and captured WNV progressive expansion from 184 predicted positive municipalities in 2014 to 2,012 in 2024 (an 11-fold increase in 11 years). Seasonal minimum temperature was the primary risk driver, followed by monitoring year and population density, indicating active spatial spread. Environmental suitability consistently preceded clinical detection. Municipalities with cases in 2023-2024 exhibited significantly higher predicted suitability during 2018-2022 than those without cases (average risk 0.58 vs 0.20). Our model successfully identified emerging risk hotspots along the Adriatic coast and southern Italy before the official human spillover of 2025. CONCLUSION Embedding macroecological drivers into WNV risk modelling provides an improved understanding of drivers of rapid WNV expansion. Our model enables proactive risk mapping, surveillance efforts, and targeted public health measures.

Summary Background: Wastewater and environmental surveillance (WES) enables community-level monitoring of infectious diseases. Most progress has focused on sewer-based surveillance, yet nearly half of the global population relies on non-sewered sanitation. In non-sewered settings, urban drainage channels have been used for poliovirus environmental surveillance, but their potential for a multi-pathogen WES with spatially defined catchments, and comparability to sewer-based surveillance, remains under-explored. Methods: Ten drainage channel sampling points with delineated micro-catchments (0.95-3.83 km2 with 12,885-44,146 people) were selected within Kampala. A total of 255 drainage channel and 54 wastewater treatment plant (WWTP) influent samples were collected during two campaigns in March and September-October 2025. A multi-target panel was quantified by digital PCR, including enteric viruses (Norovirus GI and GII, Rotavirus), respiratory viruses (SARS-CoV-2, Influenza A and B viruses, Respiratory Syncytial Virus (RSV)), non-O1/O139 Vibrio cholerae (V. cholerae), and Pepper Mild Mottle Virus (PMMoV) as a fecal indicator. Findings: Enteric viruses, non-O1/O139 V. cholerae, and PMMoV were consistently detected across all drainage channels and WWTP influents. Concentrations were generally lower in drainage than WWTP influents, except for non-O1/O139 V. cholerae. When normalized by PMMoV, concentrations across most drainage channels were comparable to WWTP influents, although comparability varied by target and location. Both concentrations and PMMoV-normalized concentrations varied across sampling locations. Trends between campaigns varied by pathogen target and were not explained by any single micro-catchment characteristic. Influenza A virus was the most frequently detected respiratory virus (8-23% in drainage channels; 3-10% in WWTP influents), while SARS-CoV-2, Influenza B, and RSV were rarely detected. Interpretation: PMMoV-normalized concentrations across most micro-catchments were comparable to WWTP influents, with spatial heterogeneity implying neighborhood-level differences in disease prevalence. Catchment-delineated drainage surveillance has potential to offer spatially resolved public health information in non-sewered settings comparable to sewer-based wastewater monitoring. Funding: Eawag Discretionary Funding

Background: In low- and middle-income countries, laboratory testing to rapidly detect measles outbreaks is limited by infrastructure availability and high costs. This study estimates the potential impact and cost-effectiveness of measles rapid diagnostic tests (RDTs) if implemented nationally in Ethiopia to either replace or expand current testing. Methods: An agent-based model to simulate measles outbreaks was calibrated to Ethiopian measles surveillance data. Modelled outbreak outcomes were aggregated over a 10-year period. Scenarios included using RDTs to (1) replace laboratory testing; (2) replace epidemiological linkage; and (3) increase case detection, in addition to replacing laboratory testing and epidemiological linkage. Testing and outbreak response costs (in 2025 US$) were obtained from Ethiopian Public Health Institute from a government perspective. Total costs and disability-adjusted life years (DALYs) for each scenario were compared to baseline. Results: All scenarios were cost saving compared to baseline. Replacing laboratory testing with RDTs saved US$4.2M (3.2M-4.9M) over 10-years, but due to very low testing rates the benefits of eliminating laboratory testing delays were offset by missed cases from the lower RDT sensitivity, leading to similar outbreak detection times and DALYs. Replacing epidemiological linkage with RDTs had similar DALYs but increased the cost savings to US$9.7M. Using RDTs to double case detection reduced outbreak detection time from 113 to 80 days, averted 17,000 DALYs, and saved US$4.3M. Conclusions: In Ethiopia, use of measles RDTs could be cost saving, and if used to expand testing could prevent measles infections through faster outbreak detection and response.

BackgroundIdentifying risk factors for Leishmania infection in dogs is essential for understanding disease epidemiology and informing control and targeted prevention. This systematic review and meta-analysis examined these risk factors across endemic and non-endemic regions of Europe. MethodsInclusion criteria included studies that quantified the association between Leishmania infection (both asymptomatic infection and canine leishmaniosis) and risk factors among European dogs. Non-peer reviewed grey literature and studies that focus only on prevention were excluded. The search included all records indexed in PubMed up to 17th March 2026. Risk of bias was assessed using the ROBINS-E tool. Random-effects multilevel models were used for meta-analysis, and sensitivity analyses were conducted to assess robustness. ResultsOf the 46 studies included, 94% were cross-sectional and 76% assessed seroprevalence. Studies were primarily conducted in Southern Europe (Spain 35%, Portugal 17%, Italy 17%, and Greece 11%). There was considerable evidence of an association between increasing age and infection (OR = 1.15 [95% CI: 1.10, 1.21] per year). Five additional factors showed some evidence of association: male sex (OR = 1.17 [1.01, 1.36]), higher body weight (OR = 1.06 [1.02, 1.09] per kg), outdoor housing (OR = 1.84 [1.26, 2.69]), outdoor sleeping (OR = 2.50 [1.52, 4.09]), and neuter status showing higher odds in neutered dogs (OR = 2.22 [1.37, 3.61]). Other potential risk factors, with weak or no clear evidence, included fur length, coat color, breed, utilization (e.g. pet or hunting), urbanicity, care setting, clinical signs, cohabitation, co-infection, sand fly density, land cover, home features, owner perception and knowledge of the disease, travel history and origin of dog, and area-level socioeconomic deprivation. Over 75% of analytical units were assessed as having a high risk of bias, with the remaining assessed as moderate risk. Inadequate handling of confounders was the primary source of bias risk. ConclusionsThe results highlight key risk factors to consider in future research and for improving prevention and disease control, with most identified factors likely acting as proxies of exposure to sand fly bites. Future studies should ensure adequate control for confounding, while causal inference would be strengthened by conducting longitudinal cohort studies.

Introduction: Veterinary Professionals are at risk of contracting zoonoses, including potential emerging infections. Personal Protective Equipment (PPE) could reduce infection transmission risk, but use of it in the profession is low. Understanding Veterinary Professionals experiences with PPE could help identify facilitators and barriers, therefore informing a future strategy to improve usage. Methods: Two focus group discussion with veterinary nurses and two with veterinarians took place at a tertiary small animal teaching hospital. With an interpretivist epistemology, transcriptions were inductively coded and analysed using thematic analysis. Results: Veterinary Professionals frequently reported underusing PPE, despite significant zoonotic risk. Friction in interdisciplinary relationships negatively impacted veterinary professionals experiences with PPE: differing views on risk, policy and PPE, contrasting perceptions of each other, and challenges with communication impacted PPE decision-making. Barriers included an absence of initial recognition of risk, a staff culture of prioritising patient health over risk to self, friction in response to others PPE use or lack thereof, an overly complex policy, and a lack of cultural and institutional reaction to occupationally contracted zoonoses. Facilitators included effective inter and intradisciplinary communication and previous personal experience with serious zoonoses. Conclusion: Veterinary Professionals experiences with PPE, are shaped by social and professional factors, such as interdisciplinary friction, perceptions of self and others, of risk and policy. Recommendations are for the findings to be used locally to embed PPE use into everyday practice. At a national level, the United Kingdom Health Security Agency (UKHSA) should use the findings to evaluate current policies regarding PPE and zoonoses in veterinary practice and produce a simplified national guidance, in collaboration with veterinary professionals with lived experience. Further research into General Practice Veterinary Professionals experiences and the interdisciplinary social dynamics is recommended.

Antimicrobial resistance is a growing threat to health systems, but stewardship programs must compete for funding with many urgent health priorities in low- and middle-income countries. Evidence that quantifies not only effectiveness but also economic value is therefore essential for policy and budget decisions. We evaluated targeted antimicrobial stewardship programs in four tertiary hospitals in Nepal and Laos using interrupted time-series analyses of antibiotic use, combined with micro-costing to estimate benefit-cost ratios. Stewardship was associated with immediate reductions in antibiotic use across the three Nepal hospitals, whereas effects in Laos were more heterogeneous. Economic returns were positive across sites, with the largest returns observed in the private hospital in Nepal. Here, we show that pragmatic, ward-focused stewardship can reduce antibiotic use and generate measurable economic value in resource-constrained hospital settings, supporting its prioritization as a scalable investment for antimicrobial resistance control.

Existing molecular diagnostic approaches for notifiable avian diseases (NADs) involve a suite of PCR assays that enable both generic detection, and where positive, subtyping of both avian influenza virus (AIV) and Newcastle disease virus (NDV). Novel rapid and direct diagnostic assays for the detection of AIV and NDV were developed and evaluated using unprocessed cloacal (C) and oropharyngeal (OP) poultry swab material. Both assays employ a closed tube direct real-time reverse transcription polymerase chain reaction (RRT-PCR) approach in which viral lysis is achieved by heat treatment and a dedicated PCR compatible buffer, followed by detection using a RRT-PCR approach. Primer and probe sets were designed using globally circulating AIV and NDV sequences collected over the preceding five years, rather than region-specific sequence datasets, so that the assays detect all circulating genotypes. Analytical performance assessment demonstrated that both assays were highly sensitive and specific, successfully detecting all unextracted target antigens without cross reactivity to a panel of other common poultry pathogens. For each assay, viral lysis and amplification were achieved directly from samples at single digit genome copy numbers. Furthermore, low levels of viral RNA could be reliably detected in the presence of C and OP matrix material, providing proof-of-concept for direct detection of these economically significant avian pathogens in a field setting. Additional use case scenarios, including pooled sample screening and combined C/OP testing from individual birds, were also explored. These findings establish a foundation for ongoing studies incorporating paired-sample testing against validated laboratory reference assays.

Background: Alpha-gal syndrome (AGS) is an emerging disease, increasingly recognized as a public health concern in the United States. The primary cause of AGS in the United States is the bite of Amblyomma americanum ticks. White-tailed deer serve both as a preferred food source and as transport for A. americanum. In this work, we aim to quantify the effect of white-tailed deer abundance on number of AGS cases in United States counties. Methods: To mitigate concerns about confounding, we used the front-door formula, leveraging biological knowledge about the causal process. Due to lack of official data, our analysis relied on data made available by citizen science efforts. Results: We found that a higher number of reported white-tailed deer sightings in 2020 was associated with the county-level presence of A. americanum in 2024. In turn, county-level presence of A. americanum was associated with a higher number of self-reported AGS cases. We estimated that if white-tailed deer abundance had increased by 50%, 75%, or 100% in 2020, there would have been 89 (95%CI: 12, 252), 126 (12, 354) or 159 (6, 448) additional AGS self-reported cases in the US in 2025. Conclusions: The estimated associations are compatible with an effect of white-tailed deer abundance on AGS in the country. Due to measurement error, the low granularity of the available data, the ecological nature of the design, and the modelling choices, our effect estimates should be interpreted cautiously. Further studies are needed to quantify the population-level effect of white-tailed deer on AGS.

Vaccination of free-roaming dogs (Canis lupus familiaris) and cats (Felis catus) remains a major public health challenge. Rapid urbanization forces these species into complex contact zones, where structural failure of pulsed vaccination under high demographic turnover undermines standard One Health interventions. In such cases species-specific intervention cycles are needed to reduce zoonotic disease risk. We integrated field-data within a simulated vaccination campaign to determine how species-specific turnover rates drive the erosion of herd immunity at an Amazonian urban sentinel site (university campus). We monitored free-roaming populations (72 dogs, 75 cats) using a non-invasive photographic mark-resight protocol from 2023 to 2025. We modelled time-to-disappearance using Cox Proportional Hazards and simulated the trajectory of effective vaccination coverage against a 40% herd immunity threshold, distinguishing between loss of vaccinated individuals and recruitment of susceptible individuals. The campus functioned as a high-turnover system, with 72% of dogs and 48% of cats classified as transients. Species significantly predicted persistence (Hazard Ratio = 0.56; 95% CI: 0.33-0.94; p=0.029), with cats exhibiting double the median residency of dogs (432 vs. 193 days). Consequently, the species experienced divergent epidemiological failure modes. For dogs, simulated vaccination coverage collapsed below a 40% herd immunity threshold in 160 days, driven by rapid immunity attrition (the loss of vaccinated individuals). Although cats persisted longer, their effective coverage was eroded by immunity dilution due to recruitment of naive juveniles, creating a 33% gap between cohort survival and population-level immunity by day 365. Annual vaccination campaigns are likely insufficient in this high-turnover urban dog population. Effective One Health zoonotic control strategies must transition from static abundance-based targets to dynamic, species-specific and turnover-adjusted intervention schedules.

BackgroundDog-mediated rabies remains a major neglected tropical disease and an important cause of preventable human mortality in sub-Saharan Africa. Despite ongoing control efforts, limited laboratory-confirmed surveillance data hinder accurate assessment of the disease burden and the development of evidence-based interventions in Ghana. This study investigated the epidemiological characteristics of laboratory-confirmed rabies cases among suspected animal submissions and evaluated the contribution of routine diagnostic surveillance to rabies control. Methodology/Principal FindingsA retrospective analysis was conducted using laboratory surveillance records of suspected rabies cases submitted to a regional veterinary laboratory in Ghana between 2020 and 2025. Brain tissue samples were examined using Sellers staining, the direct fluorescent antibody test (FAT), and polymerase chain reaction (PCR). Epidemiological information, including animal species, vaccination history, ownership status, bite history, and diagnostic outcomes, was extracted and analysed descriptively. Of the 51 suspected cases examined, 46 (90.2%) were laboratory confirmed as rabies. Domestic dogs accounted for 96.1% of submissions, while cats represented 3.9%. Most suspected animals were either unvaccinated (64.7%) or had unknown vaccination status (33.3%), and 90.2% had reportedly bitten humans before submission. Confirmed cases were detected throughout the study period, indicating sustained viral circulation and continued zoonotic risk within the study area. Conclusions/SignificanceThe consistently high proportion of laboratory-confirmed rabies cases demonstrates persistent transmission of rabies virus among domestic animals and highlights substantial risks to human health. The integration of conventional and advanced laboratory diagnostic methods strengthened surveillance and improved case confirmation. These findings underscore the urgent need to expand dog vaccination coverage, strengthen laboratory-based surveillance, improve bite reporting systems, and enhance collaboration between veterinary and public health sectors. Reinforcing integrated One Health strategies will be essential to accelerate progress towards the global goal of eliminating dog-mediated human rabies by 2030. Author SummaryRabies remains one of the worlds oldest and most devastating zoonotic diseases, which is fatal yet preventable, causing thousands of human deaths each year, particularly in Africa and Asia. In Ghana, domestic dogs are the principal reservoir of the rabies virus, but information on the epidemiology of animal rabies from routine laboratory surveillance is limited in many regions. We reviewed six years (2020 - 2025) of laboratory records from the Regional Veterinary Laboratory in Techiman, Bono East Region, to describe the epidemiological characteristics of suspected animal rabies cases and laboratory-confirmed infections. Of the 51 brain samples submitted for diagnosis, 46 (90.2%) were confirmed positive for rabies. Domestic dogs accounted for almost all submissions, and most confirmed cases involved animals that were either unvaccinated or had an unknown vaccination history. Furthermore, most suspected animals had reportedly bitten humans before laboratory investigation, highlighting the continuing public health risk posed by canine rabies. The study also documents how regional diagnostic capacity was strengthened through collaboration with national reference laboratories, enabling the routine use of internationally recommended diagnostic methods such as the fluorescent antibody test and polymerase chain reaction. These findings provide valuable evidence to support enhanced rabies surveillance, increased mass dog vaccination, improved public awareness, and strengthened One Health collaboration. Such measures are essential for reducing human rabies deaths and supporting Ghanas contribution to the global goal of eliminating dog-mediated human rabies by 2030.