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Whole-Genome Sequencing of Feline Uropathogens Reveals Multidrug Resistance and Zoonotic Potential in Domestic Cats in Tunisia.

BACKGROUND: Urinary tract infections (UTIs) in cats are increasingly recognized as clinically relevant conditions frequently associated with multidrug-resistant (MDR) bacteria of potential zoonotic origin, yet genomic data on feline uropathogens remain scarce in Tunisia. METHODS: We used whole-genome sequencing to characterize seven bacterial isolates recovered from six cats with clinical signs of UTI: Mammaliicoccus lentus (n = 2), Staphylococcus schleiferi (n = 1), Mammaliicoccus sciuri (n = 1), Enterococcus faecalis (n = 1), Enterococcus casseliflavus (n = 1), and Klebsiella aerogenes (n = 1). RESULTS: Resistome analysis revealed determinants conferring resistance to β-lactams (blaZ, blaCMY-132), methicillin (mecC-type), macrolides (erm(43), ermB), tetracyclines (tet(M), tet(45), tetB), fosfomycins (fosI, fosB, fosA5), and aminoglycosides (aac(6'), aph(3')-IIIa, aph(6)-Id), alongside efflux pump genes (efrA, sepA, sdrM, oqxA, KpnE/F/G), vancomycin-operon genes (vanT, vanY, vanC, vanG), and biofilm/biocide-tolerance genes (salB, qacG). Notably, M. lentus S104 carried mecC-type elements, the first such report in Tunisia, while K. aerogenes displayed an extensive MDR profile, including blaCMY-132 and fosA5. Multilocus sequence typing/ribosomal multilocus sequence typing (MLST/rMLST) identified diverse lineages, including the internationally distributed E. faecalis ST19 and the rarely reported K. aerogenes ST242. Plasmids were absent in all isolates; a Tn916/1545-type transposon occurred in E. casseliflavus, and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems were unevenly distributed. CONCLUSIONS: These findings highlight companion animals as reservoirs of clinically important resistance genes, reinforcing the need for One Health AMR surveillance.

Animals

Molecular Evolution and Zoonotic Potential of Muju Virus (Orthohantavirus puumalaense) in Craseomys regulus, Republic of Korea.

Orthohantavirus puumalaense causes hemorrhagic fever with renal syndrome in Europe, with Puumala virus (PUUV) as its primary representative. Muju virus (MUJV), harbored by Craseomys regulus, an Arvicolinae rodent species endemic to the Republic of Korea (ROK), is also a genotype of O. puumalaense. However, their genomic diversity and zoonotic potential remain largely unknown. To investigate their prevalence, 185 voles were collected from 23 regions of the ROK between 2012 and 2023. Serological assays detected anti-PUUV immunoglobulin G antibodies in five samples (3.1%), whereas reverse-transcription polymerase chain reaction confirmed MUJV RNA in identical specimens (2.7%). Amplicon-based nanopore sequencing facilitates near-complete genome recovery, enabling high-resolution comparative analysis. Phylogenetic analysis revealed distinct genetic lineages in Gangwon and Jeollabuk Provinces. Evolutionary rate estimates indicated greater sequence divergence in the S and L segments than in the M segment. A zoonotic risk assessment revealed that most MUJV variants exhibited moderate-to-high spillover potential. The molecular detection of MUJV in Cheorwon, Gangwon Province, expands its known geographic range and provides the first molecular evidence of MUJV circulation in this region. These findings highlight the need for continued surveillance and seroprevalence studies of MUJV to assess its potential for human exposure and public health relevance in the ROK.

Animals

Clinical and Microbiological Insights into Caseous Lymphadenitis in Sheep and Goats in Khorasan Razavi, Iran.

INTRODUCTION: Caseous lymphadenitis (CLA), a chronic bacterial disease caused by Corynebacterium pseudotuberculosis, significantly impacts small-ruminant health and productivity worldwide, causing economic losses through reduced wool and milk yields, reproductive issues, and carcass condemnation. Despite its importance, CLA prevalence and microbial dynamics remain under explored in Iran, where small ruminants are vital to rural economies. This study assessed the prevalence, clinical manifestations, and bacteriological profile of CLA in Khorasan Razavi Province, northeast Iran, to inform regional control strategies and address potential zoonotic risks. MATERIALS & METHODS: We examined 15 flocks totaling 4,733 animals (4,640 sheep, 93 goats) through clinical inspections and microbiological analysis of pus samples from affected lymph nodes. RESULTS: The results revealed a lymphadenitis prevalence of 11.59% (95% CI, 10.58%, 12.66%), with 8.62% of sheep (400/4640) and 8.60% of goats (8/93) affected, varying across flocks from 0% to 28.57%. Submandibular lymph nodes were most commonly affected (51.35%), followed by retropharyngeal (18.02%) and parotid (15.32%) nodes, with peak incidence in the 2-3-year age group (38.24%), likely linked to shearing practices. Bacteriological analysis of 102 pus samples identified C. pseudotuberculosis in 19.6% (20/102) of cases, characterized by small, dry, white colonies with β-hemolysis on Columbia blood agar. A diverse microbial profile included Actinobacillus spp. (7.8%), Trueperella pyogenes (3.9%), and novel isolates like Acinetobacter spp. and Yersinia spp. (1.0% each), with 43.14% of samples sterile, suggesting chronicity or sampling challenges. CONCLUSION: These findings indicate CLA etiology is complex, extending beyond a single pathogen and influenced by local husbandry practices. The study underscores CLA's economic burden and zoonotic potential, given rare but documented human cases. Integrated control measures-enhanced molecular diagnostics, recombinant phospholipase D (PLD) vaccine trials, and improved biosecurity-are urgently needed. Future research should prioritize genomic strain typing and environmental reservoir analysis to refine CLA management in Northeast Iran, offering insights applicable to similar agroecosystems globally.

Animals

Association of Enterocytozoon bieneusi Infection with chronic/persistent diarrhea and ITS genotypic diversity: a hospital-based case-control study in Suburban Shanghai, China.

Enterocytozoon bieneusi is a globally distributed zoonotic enteric pathogen that remains largely overlooked in routine diarrheal disease surveillance. Although previous studies in Shanghai, China, have reported elevated prevalence in diarrheal populations, case-control data from suburban areas at the peri&#x2011;urban interface and the strength of the association between E. bieneusi infection and chronic diarrhea in non-immunocompromised individuals remain poorly characterized. We performed a hospital-based case-control study in suburban Shanghai, enrolling 286 diarrheal outpatients without documented immunodeficiency and 138 asymptomatic controls frequency-matched for age and sex. Fecal specimens were collected and subjected to genomic DNA extraction. E. bieneusi was detected via nested PCR amplification of the ribosomal internal transcribed spacer (ITS) region. Factors associated with infection were identified using multivariate logistic regression. Genotypic diversity and zoonotic potential were assessed by Sanger sequencing and phylogenetic analysis. The overall prevalence of E. bieneusi was 12.2% (35/286) in diarrheal patients, significantly higher than the 2.2% (3/138) observed in asymptomatic controls (P < 0.001). E. bieneusi positivity was independently associated with chronic/persistent diarrhea (adjusted odds ratio = 2.63, 95% confidence interval: 1.25-5.54, P = 0.011). Fourteen distinct ITS genotypes were identified, comprising five known genotypes (D, EbpD, SHW7, Henan-III, and CHG5) and nine novel genotypes (designated SHH2 to SHH10). Thirteen genotypes clustered within Group 1, and one genotype (CHG5) fell within Group 2, two phylogenetic groups that contain genotypes with documented zoonotic potential in global surveillance. E. bieneusi was detected at a relatively high prevalence among diarrheal patients in suburban Shanghai, and its detection was associated with chronic/persistent diarrhea. The predominance of zoonotic genotypes and the identification of nine novel Group 1 genotypes indicate phylogenetic similarity to known zoonotic lineages and warrant further investigation of local zoonotic transmission; no animal or environmental samples were analyzed in this study. These findings suggest that E. bieneusi testing may be considered as part of the differential diagnosis for patients with unexplained chronic/persistent diarrhea and highlight the need for One Health surveillance in the surveyed area.

Diarrhea

First Report of Enterocytozoon bieneusi, Encephalitozoon spp. and Blastocystis spp. in Hair Goats in T&#xfc;rkiye.

INTRODUCTION: Blastocystis spp., Enterocytozoon bieneusi and Encephalitozoon spp. are prevalent zoonotic gastrointestinal parasites that cause severe diarrhoea and enteric diseases in humans and animals worldwide. This study investigated the molecular occurrence, genetic diversity and zoonotic potential of E. bieneusi, Blastocystis spp. and Encephalitozoon spp. in domestic hair goats in Central Anatolia. METHODS: A total of 300 faecal samples were collected from hair goats in Kayseri, Aksaray and Sivas Provinces and genomic DNA was extracted from these samples. The presence of the three pathogens was detected using PCR and nested PCR targeting the SSU rRNA for Blastocystis spp., the internal transcribed spacer (ITS) for E. bieneusi and Encephalitozoon spp., respectively. Positive PCR products were sequenced to determine the species, subtypes and genotypes of the pathogens. RESULTS: The overall prevalence rates of E. bieneusi and Blastocystis spp. were 5.3% (16/300) and 6.7% (20/300), respectively. None of the faecal samples tested was positive for Encephalitozoon spp. and no co-infections among these three pathogens were detected. The SSU rRNA sequence analysis revealed Blastocystis spp. ST10, a subtype predominantly associated with animals. Additionally, one known E. bieneusi genotype (BEB6) was identified. The BEB6 genotype falls into zoonotic Group 2 of E. bieneusi in the phylogenetic tree. CONCLUSION: Our study is the first report of Blastocystis spp. and E. bieneusi infection in hair goats in T&#xfc;rkiye, highlighting their potential role in zoonotic transmission within a One Health framework. Our results provide scientific data for the prevention and control of these two intestinal pathogens. Further investigations are necessary to better understand their genetic characteristics and zoonotic potential in T&#xfc;rkiye.

Enterocytozoon bieneusi

AI-enabled viral genomics: from virus discovery to host prediction and emerging variant forecasting.

The rapid expansion of metagenomic sequencing has generated vast repositories of viral sequence data that far outpace our capacity to interpret them using conventional approaches. Highly divergent sequences, sparse functional annotation, and taxonomically uneven sampling present fundamental challenges for reference-dependent methods, which lose sensitivity precisely for novel and understudied viruses with high public health relevance. Artificial intelligence (AI) provides a new avenue to address these challenges by enabling predictive inference from viral genomes and proteins while reducing dependence on sequence similarity. In this Review, we discuss representative advances in AI for virus discovery, taxonomic classification and functional annotation, prediction of host range and zoonotic potential, and efforts toward forecasting emerging variants. These advances are transforming viral genomics from a largely descriptive discipline into one with increasing predictive capability. We also critically assess the major challenges that constrain current approaches, including the availability of high-quality and representative datasets, rigorous model evaluation, biological interpretability and responsible governance for increasingly capable AI models.

Artificial Intelligence

Characterization of the oral microbiota and antimicrobial resistance genes in shelter dogs in Japan.

Companion animals can serve as reservoirs of antimicrobial resistance genes and zoonotic microorganisms, yet information on shelter dogs remains limited. This study characterized the oral microbiota and screened for antimicrobial resistance genes in shelter dogs in Japan. Oral swabs were collected from 81 dogs, microbial genomic DNA was extracted, bacterial communities were profiled by 16S rRNA gene amplicon sequencing, and antimicrobial resistance genes were screened by PCR. We detected genes conferring resistance to several antimicrobial classes, including &#x3b2;-lactams, tetracyclines, macrolide-lincosamide-streptogramin B, phenicols, and sulfonamides. cfxA was detected in all 81 samples, followed by sul1 (66/81), tet(M) and sul2 (65/81), floR (39/81), mecA (17/81), and erm(B) (15/81). We identified potentially pathogenic genera including Capnocytophaga, Pasteurella, Fusobacterium, Campylobacter and Corynebacterium. Microbiome analysis revealed that at the phylum level, Pseudomonadota and Bacteroidota were the most dominant, while Porphyromonas, Frederiksenia and Moraxella were the most prevalent genera. Our findings highlight that (i) the oral microbiota of shelter dogs broadly resembles that reported in companion dogs and (ii) shelter dogs represent an overlooked reservoir of clinically relevant antimicrobial resistance genes and potentially zoonotic bacteria. Therefore, it is necessary to include shelter animals in antimicrobial resistance surveillance programs to capture any potential gaps in the antimicrobial resistance prevalence in companion animals and prevent dissemination of resistant bacteria to humans following adoption of shelter dogs and cats.

antimicrobial resistance gene

MicroRNAs in Veterinary Viral Diseases: A Comprehensive Review from Molecular Mechanisms to Clinical Translation.

MicroRNAs (miRNAs) are small non-coding RNA molecules, approximately 22 nucleotides in length, that regulate post-transcriptional gene expression and have emerged as pivotal modulators of host-virus interactions. Veterinary viral diseases continue to pose substantial challenges to animal health, livestock productivity, food security, and public health, particularly due to their zoonotic potential. While miRNA research has advanced considerably, a comprehensive and critically integrated understanding of their biological functions and clinical applications across veterinary viral diseases remains incomplete. This comprehensive critical narrative synthesis addresses four overarching research questions: (1) What conserved and species-specific miRNA-mediated mechanisms govern major veterinary viral diseases? (2) What contextual factors determine antiviral vs. proviral duality? (3) To what extent do circulating miRNA signatures offer diagnostic and prognostic utility? (4) What translational barriers currently prevent clinical implementation, and how can the One Health framework help overcome them? Integrating three interconnected dimensions-molecular mechanisms, pathogen-specific responses, and translational applications-the review synthesizes evidence across PRRSV, avian oncogenic viruses (MDV, ALV), the immunosuppressive IBDV, FMD, BVDV, Ebola, Hendra, Rabies, and aquatic viral diseases. A key contribution of this review is the proposal of a four-axis contextual framework that explains the antiviral/proviral duality of miRNAs, and a 'One miRNA, One Health' convergence model with a concrete implementation roadmap. Key findings include: (a) a four-axis contextual framework (cell type, infection stage, viral strain, host-viral miRNA competition) that explains the antiviral/proviral duality; (b) virus-encoded miRNAs (v-miRNAs) as lower-risk therapeutic targets due to their absence from uninfected host genomes; (c) circulating miRNA biomarkers validated only at proof-of-concept stage (TRL 1-3), with no veterinary product yet at TRL&#x2009;&#x2265;4; and (d) zoonotic conservation of miR-155, miR-146a, miR-21, and miR-122 across human and veterinary pathogens, supporting a 'One miRNA, One Health' convergence strategy. Critical short-term priorities are standardized pre-analytical protocols, open-access veterinary miRNA databases, and multicenter validation in natural infection cohorts.

Antiviral therapy

Genomic diversity and resistance determinants of staphylococci from cow and buffalo milk.

BACKGROUND: Staphylococci are important mastitis pathogens in dairy animals and serve as reservoirs of antimicrobial resistance genes (ARGs) having zoonotic potential. Genomic characterization of resistant isolates is essential to understand their diversity, resistance mechanisms, and One Health implications. METHODS AND RESULTS: A total of 363 cow and buffalo milk samples-including 108 from animals with mastitis-were screened, yielding 98 staphylococcal isolates, comprising 20 Staphylococcus aureus and 78 coagulase-negative staphylococci (CoNS). Antimicrobial susceptibility testing revealed resistance to cefoxitin (CoNS: 21.7%; S. aureus: 10%), tetracycline (CoNS: 19.2%; S. aureus: 10%), erythromycin (CoNS:16.7%; S. aureus: 10%), gentamicin (CoNS: 10.2%; S. aureus: 10%) and fluoroquinolone (CoNS: 10.2%), while the majority were sensitive to chloramphenicol, cotrimoxazole (~&#x2009;95%, each), linezolid (~&#x2009;97%), and vancomycin (100%). Nineteen isolates, including two S. aureus, were cefoxitin-resistant, and eight carried the mecA gene. Whole genome sequencing of these eight isolates revealed genome sizes ranging from 2.27 to 2.78&#xa0;MB, with the methicillin resistant S. aureus (MRSA, ERSST98) isolate possessing the largest genome and the highest rRNA copy number. Comparative genomic analysis revealed various SCCmec types along with an extensive array of resistance determinants, encompassing aminoglycosides, macrolides, tetracyclines, efflux systems, and heavy metals, underscoring the multifaceted resistance repertoire of these strains. Virulence profiling of ERSST98 demonstrated a broad arsenal of adhesins, toxins, and biofilm&#x2011;associated genes, highlighting its pathogenic capacity. Mobile genetic elements with diverse plasmid replicons and insertion sequence families further contributed to genomic plasticity. CONCLUSIONS: Collectively, this study underscores the genomic diversity of methicillin-resistant staphylococci from dairy animals with extensive resistance determinants and highlights their zoonotic relevance within One Health framework.

Animals

Toxoplasma gondii IgG seroprevalence in Mauritanian dromedary camels: First multi-regional survey.

Toxoplasma gondii is a globally distributed zoonotic parasite, and dromedary camels are important intermediate hosts in arid and semi-arid regions. However, information on T. gondii exposure in camels is lacking in Mauritania, which harbors one of the largest camel populations in West Africa. This study reports the first multi-regional seroepidemiological survey to estimate T. gondii seroprevalence and identify associated risk factors in Mauritanian dromedaries. Between 2023 and 2024, serum samples were collected from 953 camels across eight climatically distinct regions. Anti-T. gondii IgG antibodies were detected using the Modified Agglutination Test (MAT; cutoff&#x2265;1:20). Risk factors investigated included geographical region, sex, age group, and season of sampling, using multivariable logistic regression and a mixed-effects linear probability model accounting for regional clustering. The overall seroprevalence was 15.0% (143/953). Exposure varied markedly across regions, ranging from 0% in the hyper-arid northern regions of Adrar and Tagant to 41.7% in the southern Sahelian region of Guidimakha. This pronounced spatial gradient is consistent with contrasting climatic and ecological conditions, as higher rainfall and humidity in the south are hypothesized to favor environmental oocyst survival compared to the extreme aridity of the north. Geographical region and age were independent predictors of seropositivity. Compared with camels from Nouakchott, those from Guidimakha had higher odds of exposure (aOR = 2.63), whereas camels from Trarza had a markedly lower risk (aOR = 0.09). Camels older than 6 years were more than twice as likely to be seropositive as those aged 3-5 years, whereas sex and season were not associated with seropositivity. These findings indicate that T. gondii exposure is widespread in Mauritanian dromedaries and that ecological conditions may influence exposure patterns. The marked spatial heterogeneity supports targeted surveillance and One Health interventions to reduce the potential zoonotic risk associated with camel-derived food products.

Animals

Genomic evidence of active circulation of Orthobunyavirus in Ecuador.

BACKGROUND: Between 2023 and 2025, the largest Oropouche fever epidemic recorded in history unfolded across Brazil seeding cases throughout Latin America. In 2024, three cases of Oropouche fever were reported in Ecuador. METHODS: An Oropouche fever case detected in Bol&#xed;var Province in June 2024 was preliminarily diagnosed as Oropouche virus (OROV) through RT-qPCR and was further processed using next-generation sequencing. RESULTS: Segments L and S of this virus forms a monophyly with another sequence circulating in Ecuador in April 2024 (i.e. PQ863772.1 isolate Ecuador traveler), with an uncertain province origin. Both sequences differ from previous OROV Ecuadorian sequences detected in 2016 and from the OROV strain driving the 2023-2025 epidemic in Brazil. CONCLUSIONS: Orthobunyavirus oropoucheense has an endemic circulation in Ecuador. Genomic surveillance of Orthobunyavirus in Ecuador and other regions should be actively pursued-independent of epidemics-to anticipate potential zoonotic outbreaks.

Adult

Population structure and antibiotic resistance of Salmonella isolates from diseased poultry in Jiangxi Province, China.

Salmonella poses a significant threat to human and animal health. However, the relationship among population diversity, antibiotic resistance, and infection risk remains largely unexplored. In this study, 69 Salmonella strains were isolated from diseased poultry in Jiangxi Province from 2021 to 2024. Using whole-genome sequencing, serotype prediction, MLST, virulence and resistance gene analysis, antibiotic susceptibility testing, and mobile genetic element annotation, we characterized the diversity, resistance profiles, and transmission mechanisms of these strains. The results showed high diversity, with Salmonella enterica subsp. enterica serovar Typhimurium (>60%) and ST19 (62.31%) as the dominant serovar and sequence type, respectively. Several avian isolates were genomically similar to human isolates, indicating potential zoonotic risk. All strains harbored conserved core virulence modules, whereas accessory modules (e.g., cdtB, astA, pefA) varied and may affect pathogenicity. The multidrug resistance rate was 97.1%, with 100% resistance to erythromycin, tilmicosin and tiamulin, and resistance rates of 91.3%, 84.1%, and 71.0% to sulfonamides, enrofloxacin, and ceftiofur, respectively. Sixty-eight resistance genes were identified. Highly conserved antimicrobial resistance gene (ARG) modules (e.g., sul2-aph(3&#x2033;)-Ib-aph(6')-Id-tet(A)) were shared between chromosomes and plasmids and were flanked by mobile elements such as Tn3 and IS3. Genomic islands (GIs) and plasmids in some strains carried resistance gene clusters highly homologous to those in pathogens from humans, pigs, and chickens, suggesting active horizontal transfer of resistance genes across hosts. This study revealed high diversity, prevalent multidrug resistance, and active horizontal transfer of resistance genes in avian-derived Salmonella from Jiangxi Province, emphasizing the need for cross-host resistance monitoring and antibiotic management within the 'One Health' framework.

Horizontal gene transfer

Circulation of avian Chlamydia abortus in the Netherlands and community-acquired pneumonia: an outbreak investigation and retrospective cohort study.

BACKGROUND: In 2021, a novel group of Chlamydia strains in wild birds was classified as avian Chlamydia abortus, with unknown zoonotic potential. We report relevant features of avian C abortus infections from a Dutch family cluster and unrelated historical cases using clinical, epidemiological, and microbiological data. METHODS: An outbreak of avian C abortus started in the Netherlands in December, 2022. Source investigation was done using questionnaires to interview patients and environmental sampling. The outbreak strain of avian C abortus was cultured from three patients from whom sufficient material was available for culture and underwent whole-genome analysis. The outbreak strains and retrospective cohort study strains previously submitted to the National Human Psittacosis surveillance programme in the Netherlands between 2010 and 2022 were typed by partial ompA sequencing. Strains with the same aberrant ompA genotype were further analysed with XerC gene plasmid analysis and compared with closely related Chlamydia sequences available in GenBank. FINDINGS: An avian C abortus strain caused a cluster of respiratory illness in four family members. Three patients were hospitalised with community-acquired pneumonia, one of whom was admitted to the intensive care unit. The faeces of wild birds were considered a probable source for the index infection. For two family members, human-to-human transmission was a plausible route. Ten historical cases could be identified with avian C abortus with the same ompA genotype. All patients had been admitted to hospital, at least five developed pneumonia, and one died. INTERPRETATION: This cluster supports that avian C abortus strains can cause human infections and underlines that human-to-human transmission should be considered when tracing the source of such infections. FUNDING: National Institute for Public Health and the Environment and Dutch Ministry of Agriculture, Fisheries, Food Security and Nature. TRANSLATION: For the Dutch translation of the abstract see Supplementary Materials section.

Humans

Characterization and evolutionary history of novel SARS-CoV-2-related viruses in bats from Cambodia.

Circulating bat coronaviruses present a significant pandemic threat, yet our understanding of their genetic diversity and evolutionary dynamics remains limited. Over 3 years, we sampled 1,462 bats in Cambodia's Steung Treng province, identifying extensive and diverse coronaviruses co-circulation. Using metatranscriptomic and amplicon sequencing, we generated 33 complete sarbecovirus genomes sequences, revealing novel lineages that cluster into four distinct groups, each associated with different Rhinolophus bat species. Our analysis highlights rapid migration and recombination of sarbecovirus lineages over short distances and timescales. Of note, the receptor-binding domains of two novel viral groups exhibit high similarity to SARS-CoV-2, and pseudovirus assays confirmed the ability of this spike protein to mediate entry into cells expressing human ACE2, suggesting a potential zoonotic risk. The observed genetic diversity underscores the urgent need for continuous surveillance to identify high-risk animal-to-human interfaces and inform pandemic preparedness.

Animals

Performance of the IR Biotyper, Nanopore, and Illumina sequencing to discriminate Escherichia coli strains originating from poultry.

UNLABELLED: Escherichia coli is a highly diverse bacterial species that includes avian pathogenic E. coli (APEC), one of the most prevalent causative agents of disease in poultry worldwide. Rapid and accurate discrimination of E. coli strains is essential for outbreak management, antimicrobial resistance surveillance, and vaccine development. In this study, we compared the performance of Fourier Transform Infrared (FTIR) spectroscopy using the IR Biotyper system with Nanopore and Illumina whole-genome sequencing (WGS) for typing 200 E. coli isolates, originating from four poultry rearing farms in the Netherlands. From each farm, we sampled 10 one-day-old meat type rearing chicks, and from every chick, we isolated 5 E. coli strains. FTIR clustering showed strong concordance with WGS-based classifications, particularly serotyping and core-genome similarity determined by PopPUNK analysis (Adjusted Rand Index 0.75-0.92). While Nanopore and Illumina sequencing provided the highest genetic resolution, FTIR offered a faster (max 6 vs 12-28 days for 200 isolates) and more cost-effective alternative for assessing clonality. Across all methods, multiple strains were detected per farm, whereas most birds carried a single dominant E. coli strain. Our findings demonstrate that FTIR provides a reliable and scalable phenotypic method for rapid strain discrimination in E. coli, complementing WGS in diagnostic, surveillance, and epidemiological settings where speed and throughput are critical. IMPORTANCE: Escherichia coli is a major pathogen in poultry and a potential zoonotic risk for humans. Rapid and accurate discrimination of avian pathogenic E. coli (APEC) strains is critical for outbreak management, antimicrobial resistance surveillance, and the design of effective autogenous vaccines. In this study, we compared Fourier Transform Infrared (FTIR) spectroscopy with Nanopore and Illumina whole-genome sequencing for strain typing of E. coli isolates originating from poultry. The results show that FTIR provides comparable clustering accuracy to genomic approaches at a fraction of the time and costs. This work demonstrates that FTIR can serve as a practical, high-throughput alternative for routine monitoring of E. coli in veterinary diagnostics and food safety of poultry meat, enabling faster decision-making and more targeted interventions across the poultry production chain.

Animals

Prevalence of Erysipelothrix rhusiopathiae in tonsils of domestic pigs and wild boars in Sweden.

Erysipelothrix rhusiopathiae (ER) causes erysipelas in multiple animal species and may persist in the environment or be carried asymptomatically. It is estimated that 30-50% of apparently healthy or convalescent pigs harbour ER in their tonsils and other lymphoid tissues. This study aimed to determine the prevalence of ER in the tonsils of healthy Swedish fattening pigs and wild boars. Tonsils were collected from 200 fattening pigs at slaughter from ten abattoirs across Sweden in 2017, with one pig per herd sampled. Wild boars (n&#x2009;=&#x2009;180) were sampled during hunting, primarily in &#xd6;sterg&#xf6;tland County, in 2018. Cultures were performed using selective media and isolates were confirmed as ER by MALDI-TOF MS. ER was recovered from 6/200 pig tonsils (3.0%), all originating from three abattoirs in southern Sweden. ER was isolated from 76/167 (45.5%) of wild boar tonsils. Whole-genome sequencing revealed a high genetic diversity among the isolates with no dominant clones. Overall, these results indicate that Swedish pig husbandry, characterized by indoor rearing of fattening pigs, age-segregated rearing, sow vaccination, enhanced biosecurity, and restricted straw access largely prevents tonsillar colonization by ER aligning with the low occurrence of clinically diagnosed erysipelas in such herds. For wild boars, the high isolation rate suggests that wild boar could act as a reservoir and potential source of infection for domestic pigs. The potential zoonotic risk should also be considered.

Animals

Influenza A virus in Swiss pig herds with respiratory disease: Seasonality and age dependence.

Influenza A virus (IAV) is an important respiratory pathogen in pigs and poses a zoonotic risk to humans in close contact. While IAV epidemiology has been extensively studied in large-scale production systems, data from Switzerland - characterized by small herds and limited live pig imports - remain scarce. This exploratory nationwide cross-sectional study aimed to assess the association between herd-level IAV detection and reported respiratory disease in pig herds, and to explore associations with husbandry-, animal-, and human health-related factors. Between November 2023 and April 2025, 25 Swiss pig herds with caretaker-suspected respiratory symptoms were investigated. In each herd, five nasal swabs were collected and analyzed by quantitative PCR. Herd managers completed an interview, and clinical examinations were performed. Overall, 56&#x2009;% (95&#x2009;% CI: 37,1 - 73,3) of herds tested positive for IAV, comparable to reports from other European countries. The estimated intra-herd detection rate was 49,6&#x2009;% (95&#x2009;% CI: 31,2 - 68,0). Respiratory disease outbreaks associated with IAV detection showed indications of seasonal variation, with no positive herds identified during summer. Across age groups, pigs aged 11-14 weeks had a higher likelihood of IAV detection, with 15,79-fold increased odds (95&#x2009;% CI: 1,50 - 860,4), although with considerable uncertainty. The interpretation is limited by the small sample size, heterogeneous data, and reliance on single time-point qPCR detection. The results suggest that IAV detection in clinically apparent respiratory outbreaks may follow seasonal patterns in Swiss pig herds. Weaners and newly introduced fattening pigs may play a role in such respiratory outbreaks and could represent relevant targets for IAV surveillance in Switzerland. Continued monitoring and the implemen tation of appropriate control measures remain important given the virus's zoonotic potential and impact on pig health.

Animals

Coronavirus surveillance in passerines reveals novel deltacoronaviruses in Eurasian tree sparrows with implications for One Health and livestock biosecurity.

Coronaviruses (CoVs) are widespread RNA viruses infecting a broad range of avian and mammalian hosts. Although gammacoronaviruses and deltacoronaviruses (DCoVs) are common in wild birds, their presence in Eurasian passerines remains poorly understood. We screened 243 birds (35 species) at migratory stopover sites in Slovenia (2020-2021) using pan-coronavirus RT-PCR. Coronavirus RNA was detected only in four Eurasian Tree Sparrows (Passer montanus). Whole-genome sequencing yielded genomes of 26,017-26,018&#xa0;bp with high internal conservation (99.95-99.98% identity). Phylogenetic analysis revealed notable evolutionary incongruence: isolates were highly related to porcine DCoVs in the ORF1ab region (95.6-96.1% amino acid identity) but clustered with divergent avian DCoVs in the spike gene (75.7-76.8% identity). RDP5 analysis provided strong evidence for a large-scale recombination event (p&#xa0;=&#xa0;1.17&#xa0;&#xd7;&#xa0;10-43), consistent with a mosaic genomic architecture combining an ORF1ab region closely related to porcine DCoVs with an avian-associated spike gene. This genomic pattern highlights evolutionary connectivity among DCoVs associated with different host groups and the potential role of recombination in changes in host association. The synanthropic behaviour and mobility of P. montanus facilitate contact with diverse hosts, making this species relevant for investigating DCoV ecology at wildlife-livestock interfaces. These findings represent the first genomic characterisation of DCoVs in P. montanus in Europe and support the inclusion of passerines in broader coronavirus surveillance. Genomic surveillance of underrepresented wild-bird hosts can improve our understanding of DCoV diversity, recombination, and evolution across wildlife-livestock interfaces.

Cross-species transmission