One of the most serious bacterial pathogens of Western honey bees (Apis mellifera Linnaeus [Hymenoptera: Apidae]) is Melissococcus plutonius, the cause of the disease European foulbrood. Because European foulbrood is highly variable, with diverse outcomes at both the individual and colony levels, it is difficult to diagnose through visual inspection alone. Common lab diagnostic techniques include microscopic examination and molecular detection through PCR. In 2009, a lateral flow device was developed and validated for field diagnosis of European foulbrood. At the time, M. plutonius was thought to be genetically homogenous, but we have subsequently learned that this bacterium exists as multiple strains, including some strains that are classified as ‘atypical’ for which the lateral flow device is potentially less effective. These devices are increasingly used in the United States, though they have never been validated using strains from North America. It is essential to validate this device in multiple locations as different strains of M. plutonius circulate in different geographical regions. In this study, we validate the field use of the lateral flow device compared to microscopic examination and qPCR on larval samples from 78 commercial honey bee colonies in the United States with visual signs of infection. In this study, microscopic diagnosis was more sensitive than the lateral flow device (sensitivity = 97.40% and 89.47%, respectively), and we found no false positive results with the lateral flow device. We find high concurrence between the three diagnostic techniques, and all three methods are highly sensitive for diagnosing European foulbrood.

Amblyomma americanum (Linnaeus) (Acari: Ixodidae) (lone star tick) is an aggressive, generalist parasite that vectors numerous important human and animal pathogens. In recent decades its geographic range has expanded northwards from endemic regions in the southeastern and southcentral United States. In 2019 five questing A. americanum ticks, comprising two life stages were detected at one site in southwestern Michigan, satisfying one CDC criterium for an established population for the first time in recent history in the state. To better characterize the extent of emerging A. americanum, we conducted active surveillance (i.e., drag sampling) in summer 2020 throughout Michigan’s southern counties and detected one adult A. americanum from each of six widespread sites, including where they had been detected in 2019. A larger established population was identified at another site in Berrien County, which yielded 691 A. americanum comprising three life stages, and questing phenologies here were similar to that reported for other endemic regions. Statewide surveillance in 2021 revealed no A. americanum outside of Berrien County, but establishment criteria were met again at the two sites where established populations were first detected respectively in 2019 and 2020. These observations may represent the successful invasion of A. americanum into Michigan. Data from passive (1999–2020) and active surveillance (2004–2021) efforts, including a domestic animal sentinel program (2015–2018), are reported to provide context for this nascent invasion. Continued active surveillance is needed to help inform the public, medical professionals, and public health officials of the health risks associated with this vector.

Background and Aim: Poultry is becoming an increasingly important source of protein in the Nepalese diet. The Chitwan region of Nepal is the hub of the emerging poultry industry. Little is known about the prevalence of non-typhoidal Salmonella (NTS) on poultry farms or the role of farm management practices that may contribute to the presence of NTS on farms. The role of poultry in the transmission of Salmonella enterica to humans is also poorly defined. This descriptive study seeks establish baseline data through estimation of the prevalence of NTS on broiler and layer operations in various farms of the Chitwan district of Nepal. Materials and Methods: Based on district documents on poultry production and meat marketing, a purposive sampling of 18 commercial poultry farms comprising ten broilers farms and eight layers farms was conducted. Environmental samples including water, litter, feces, feed, farm, and eggshell swabs were randomly collected from each farm. Samples were cultured and tested for the presence of NTS; positives were serotyped, and antimicrobial susceptibility determined. A comprehensive farm and practice questionnaire was administered to each farm manager. Results: The farm level point prevalence rate was 55% (10 of 18 farms) for S. enterica. Of the total 288 farm environmental samples collected, 26 samples (9%) were positive. The rate of isolation varied according to the origin of samples: Water (27.5%), feces (10.6%), litter (8.6%), farm swabs (5%), feed (1.8%), and eggshells (0%). Farm management variables/risk factors are summarized and categorized as non-modifiable and modifiable for analysis. Broiler operations were more likely to be positive than layer operations as were poultry houses with two or less open sides. All-in/all-out management style was found to be protective. Due to the small sample size (18 farms), no associations reached statistical significance. Conclusion: Based on environmental sampling results, NTS is highly prevalent on the poultry farms in the Chitwan district of Nepal. Certain risk factors are associated with finding NTS on farms. Our findings are generally in agreement with other studies in similar countries with rapidly emerging poultry industries. The identification of risk factors provides owners, technicians, and veterinarians with some guidance to help reduce the prevalence of NTS on farms. This baseline data are critical to understanding the epidemiology of zoonotic strain of NTS in the region and are necessary for the design of future studies and mitigation plans and underlines the need for a one-health approach to protect public health-related to Salmonella spp. from poultry farms.

Background and Aim: Antibiotic-resistant infections are one of the leading threats to public health globally. Indiscriminate use of antibiotics in food animal production is an important driver of resistance, particularly among foodborne pathogens such as non-typhoidal Salmonella enterica (NTS). While there has been extensive research on antimicrobial-resistant (AMR) S. enterica in India and China, there have been few studies in countries in South Asia, including Nepal. This is particularly important with the rise of commercial poultry farming in Nepal as a means of economic development and nutritional subsistence. This descriptive study seeks to identify the prevalence and resistance patterns of NTS serotypes focusing on Chitwan, Nepal's leading poultry producing district. Materials and Methods: A mixture of purposive and judgment sampling of 18 poultry farms and 20 slaughterhouses representing a broad geographic distribution across multiple municipalities in Chitwan was conducted in May 2019. Environmental samples taken from poultry farms included: Water, litter, feces, feed, farm swabs, and eggshell swabs. Biological samples taken from nearby slaughterhouses included: Muscle, heart, liver, skin, cecum, crop, and spleen. Samples were cultured and tested for the presence of NTS. Positive isolates were serotyped and tested for antimicrobial susceptibility to seven antibiotics known to be important to both human and animal health regionally. Farm practices were also characterized through a survey, the results of which are detailed in the accompanying paper. Results: Out of 708 samples (288 environmental and 420 biological), 103 (15%) tested positive for NTS (9% of environmental; n=26, 18% of biological; n=77). The percentage of positive environmental and biological samples varied by source. Environmental sample positive rates were water (27.5%), feces (10.6%), litter (8.6%), farm swabs (5%), feed (1.8%), and eggshells (0%). Biological sample positive rates were skin (28%), heart (23%), crop (20%), muscle (15%), liver (15%), spleen (15%), and cecum (12%). Out of 103 positive S. enterica isolates, 48.5% were identified as Salmonella Typhimurium, 35% Salmonella Enteritidis, 7.8% Salmonella Gallinarum, 4.9% Salmonella Virchow, and 3.9% were Salmonella Agona. Of the 103 positive isolates, 80 (78%) were resistant to at least one antibiotic, and 21 (20%) were multidrug-resistant (MDR). Conclusion: NTS is highly prevalent among Chitwan's growing poultry industry with higher rates of positivity found in slaughterhouse samples compared with environmental samples from farms. In addition, a high rate of AMR (78%) was revealed, and an extremely concerning number of those were shown to be MDR (20%). This baseline data has important implications for poultry production and consumption in the region. Further research will elucidate the extent to which this contamination and drug resistance is impacting the health of the local population and help inform treatment and management strategies. Note: The characterization of the poultry industry and practices that might be linked to NTS contamination in the Chitwan district are detailed in the previous paper in this series (www.veterinaryworld.org/Vol.14/February-2021/14.pdf).

European foulbrood (EFB) is a severe disease of honey bee (Apis mellifera) larvae caused by the bacterium Linnaeus [Hymenoptera: Apidae]) Melissococcus plutonius (ex White) Bailey and Collins (Lactobacillales: Enterococcaceae). Many beekeepers in North America report severe EFB following blueberry pollination, but it is not clear what factors during pollination are related to clinical disease. Additionally, the impact that other factors such as viral load and hygienic behavior have on EFB has not been studied. In Spring of 2020 we enrolled 60 commercial honey bee colonies in a prospective cohort study. Colonies were inspected 3 times over the season with hive metrics and samples taken for viral testing. Each colony was tested for hygienic behavior twice and the score was averaged. Viral loads were determined by qPCR for deformed wing virus (DWV) A and B. We found no statistical difference in the EFB prevalence or severity between the 2 yards at any timepoint; 50% (n = 16) of the colonies in the holding yard and 63% (n = 17) in blueberry developed moderate to severe EFB over the study period. When colonies from both yards were pooled, we found no relationship between viral load or hygienic behavior and development of EFB. These results suggest that other factors may be responsible for driving EFB virulence and hygienic behavior is not likely helpful in managing this disease.

European foulbrood (EFB) is a severe bacterial disease of honey bee brood often leading to significant declines in colony health and honey production. The dearth of data on this disease in the United States (US) complicates response efforts. In 2021 and 2022 we conducted a two-year cross-sectional surveillance study among Michigan beekeepers to establish baseline pathogen and disease prevalence. We combined this surveillance with molecular epidemiology to investigate genetic diversity, and transmission dynamics of Melissococcus plutonius, the causative agent of EFB, in US honey bee colonies. PCR screening detected M. plutonius in all 15 migratory and stationary beekeeping operations and in 6 of 14 hobby beekeeping operations. Infection and disease were found to be seasonal, with prevalence of both peaking in June when over half the colonies were infected, and over 20% had clinical EFB. Whole genome, single nucleotide polymorphism analysis revealed wide genetic diversity even within a single hive. Operations often had multiple genotypes present which varied from year to year, consistent with high rates of transmission and reinfection. Prevalence and whole genome data provided here will be critical in tracking the efficacy of mitigation efforts and underscore the necessity of additional epidemiological investigations.