Transboundary and Emerging Diseases

Hedgehog (Erinaceus europaeus) declines in western Europe have been associated with the emergence of Hedgehog diphtheric disease (HDD), with a probable multifactorial, yet unidentified etiology. We used metagenomic sequencing of cell-free DNA (cfDNA) in hedgehog blood to identify possible causes of HDD. We detected a novel circovirus species in the European hedgehog, providing the first record of a circovirus within the mammalian order Eulipotyphla. The novel circovirus genome exhibits the characteristic circovirus structure, including a functional replicase (REP) and capsid (CAP) gene. Phylogenetic analysis placed all four detected genomes in a monophyletic clade, most closely related to sequences isolated from dogs. Subsequent PCR-based screening of 188 hedgehog liver samples demonstrated a high prevalence (61%) of this circovirus in hedgehogs brought to wildlife rescue centers, however, without any significant association with HDD. Since circoviruses are well known to interfere with host immunity across mammalian and avian taxa, the high level of circovirus detection in hedgehogs warrants further research into the role of this novel virus in hedgehog health.

Pangolins have been found to carry severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-related coronaviruses. In light of this discovery, interest has been piqued in viromes of these heavily trafficked wild animals. In this study, we performed viral metagenomic sequencing to explore viromes of both confiscated dead pangolins and captive healthy pangolins. Sequence reads of vertebrate-associated viruses in Herpesviridae, Retroviridae, Iridoviridae, Reoviridae, Arenaviridae, and Flaviviridae were detected in confiscated dead pangolins. A novel rotavirus (RV) (Reoviridae), showing a high degree of genetic similarity to the RV species D (RVD) that was previously unreported in mammals, was further confirmed by using reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing. Three out of 18 samples from the confiscated dead pangolins were positive for genomic sequences of the novel RV. Importantly, sequence alignments and phylogenetic analyses demonstrated that these RV strains genetically belonged to the RVD. Nevertheless, these novel RVD strains were divergent from known RVD strains that have been found only in Avian. They formed a separate genetic cluster. Five serial passages were attempted to isolate the RV, but no live virus was obtained. In addition, fecal samples were collected from healthy pangolins (n = 41) in our institution and screened for RVs by viral metagenomic sequencing and RT-PCR. In these fecal samples, neither species D nor previously identified species A RVs were detected. This study reported RVDs in pangolin samples for the first time to our knowledge. Identifiability disagreements between wild and captive pangolins highlight the need for further exploration into pangolin viruses to better understand their emergence and transmission potential.

Rotaviruses are well-recognized pathogens responsible for diarrhea in humans and various animal species, with Rotavirus A the most often detected and most thoroughly described. Rotaviral disease is an important concern in pathology of pigeons as well, as pigeon rotavirus A was proven to play a major role in young pigeon disease (YPD). However, rotaviruses of other groups have been so far understudied in birds. This paper describes the first finding of Rotavirus G in domestic pigeon in Europe, as well as the recovery of three complete genomes of pigeon rotavirus A with Oxford Nanopore Sequencing. Quantification of pigeon rotavirus A genetic material with droplet digital polymerase chain reaction (PCR) in pigeons suffering from diarrhea and in asymptomatic pigeons was also performed in the frameworks of this study and resulted in determination of statistically highly significant differences between the groups in both detection rate and shedding of the virus. Phylogenetic analysis revealed the close relationship of acquired strains with those originating from pigeons from Europe, North America, Asia, and Australia, indicating a broad geographical spread of pigeon rotaviruses. Results of our research shed more light on occurrence and diversity of Rotavirus species in pigeons.

The susceptibility of various animal species to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has been studied extensively. Cats have garnered significant concern due to their high susceptibility and proximity to humans. This study aimed to evaluate the susceptibility and antibody response in house cats exposed to SARS-CoV-2 when human infection was spreading in the Lisbon area during the 2019–2021 period. A total of 733 serum samples were collected and characterized from cats admitted to the Veterinary Teaching Hospital of the Faculty of Veterinary Medicine at the University of Lisbon (HEV-FMV-ULisboa). All samples were tested by enzyme-linked immunosorbent assay (ELISA) for immunoglobulin M (IgM) and immunoglobulin G (IgG) anti-SARS-CoV-2 whole Spike and receptor-binding domain (RBD) proteins from the Wuhan-Hu-1 isolate and 14.7% (108/733) tested positive, suggesting exposure to the human virus. Surrogate virus neutralization test (sVNT) against the Wuhan-Hu-1 isolate showed that 20.4% of ELISA positive samples (22/108) harbored neutralizing antibodies against the virus. The 22 most promising serum samples were retested using ELISA and sVNT against Alpha, Delta, and Omicron SARS-CoV-2 variants. Notably, these samples exhibited antibodies that were capable of recognizing and neutralizing these variants. Subsequent neutralization assays confirmed that the serum samples effectively inhibited the infection process of Wuhan-Hu-1 D614G, Delta, and Omicron SARS-CoV-2 pseudotyped viruses. Our findings indicate that cats were exposed to SARS-CoV-2 infection during the pandemic period and generated highly effective and broadly neutralizing antibodies against the virus. Although cats have not been demonstrated to significantly contribute to the spread of SARS-CoV-2, their high susceptibility to asymptomatic infection underscores the importance of investment in preventive surveillance measures. In summary, our study reinforces the notion that cats naturally infected with SARS-CoV-2 represent a valuable anthroponotic disease model in house settings and might be a potential source for the development of antibodies against SARS-CoV-2 in tackling future outbreaks with a One Heath perspective.

The emergence and evolution of avian influenza A viruses (AIVs) pose significant challenges to both public health and animal husbandry worldwide. Here, we characterized a novel reassortant highly pathogenic avian influenza virus (HPAIV), clade 2.3.4.4b H5N6, that was isolated from a mandarin duck in South Korea in December 2023. Phylogenetic and molecular analyses show that the hemagglutinin (HA) gene of the 23-JBN-F12-36/H5N6 virus clustered with HPAIV clade 2.3.4.4b H5N1 viruses, which were circulating in South Korea and Japan in 2022–2023. The M and polymerase acidic (PA) genes also revealed a close association with the HPAIV clade 2.3.4.4b H5N1 AIV that was identified previously in South Korea during November 2022. Notably, the neuraminidase (NA) gene of the 23-JBN-F12-36/H5N6 virus was estimated to have its origins in the HPAIV clade 2.3.4.4h H5N6 prevalent in poultry in China, and it is clustered with the AIVs that are associated with human infection cases. Taken together, these results show that the virus has been produced by reassortment with H5N1 HPAIV, which is prevalent in wild birds; H5N6 HPAIV, which is circulated in poultry in China; and the internal genes of low pathogenic avian influenza viruses (LPAIVs). In light of the reassortment of HPAIVs circulating in existing wild birds and HPAIVs circulating in poultry in China within the 2.3.4.4b H5Nx clade, it is imperative to strengthen active surveillance across wild bird populations, poultry farms, and live poultry markets, and to inform for the effective design of improved prevention and control strategies.

The Democratic Republic of Congo (DRC) faces a syndemic of infectious diseases, including monkeypox (mpox), cholera, measles, anthrax, and plague, worsening public health challenges and socioeconomic disparities. This review synthesizes and discusses epidemiological data and consequences of simultaneous outbreaks in the DRC between January 2023 and March 2024. The findings highlight a 6.7% fatality rate and 3319 confirmed cases of mpox, with significant outbreaks in Kinshasa and 22 other provinces. Anthrax occasionally surfaced among cattle-raising villages, measles affected fewer than five children susceptible to the disease, and cholera outbreaks persisted in North Kivu, South Kivu, and Tanganyika. Plague incidences, mostly bubonic, have been reported in Ituri province. Vulnerable groups, including children, mothers, the elderly, and those with compromised immune systems, face increased risks due to poor healthcare access, hunger, and underlying medical conditions. Cultural beliefs, healthcare system issues, and socioeconomic instability impede effective response tactics. This strain on the fragile healthcare system highlights the need for increased surveillance, immunization efforts, and community involvement. To mitigate the effects of syndemic outbreaks, strengthening the DRC’s health systems through international cooperation, integrated public health initiatives, and improved access to healthcare is crucial.

Despite significant advances in swine biosecurity (BS) over the last decade, BS plans have yet to be broadly adopted on swine farms. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScRs) framework was followed to review the literature, describe the worldwide distribution of publications on swine farm BS, and characterize the research methodologies used. The final data extraction and analysis included 157 publications originating from 48 countries. Several publications (n=93) used face-to-face interviews for data collection. An increase in the adoption of online and multimode approaches was detected after 2009. Many publications (n=92) focussed on the impact of BS on the incidence of swine diseases such as porcine reproductive and respiratory syndrome (PRRS) and African swine fever (ASF). Only 16 studies reported proposing incentives for study participation. Regions with high publication numbers were detected in Western and Southern Europe, Northeast of South America, and East Africa. Areas with low publication numbers were in Eastern Europe, North and Central Africa, Central America, and the Northwest of South America. This study identified the most common study methodologies used to assess swine farm BS. Countries with limited swine BS research studies were identified where future investigations are needed.

Nipah virus (NiV) is an emerging bat-borne zoonotic virus that can be transmitted to humans and other animals through infected bats or contaminated foods. The disease is highly lethal in humans (40%–75%) and has the potential for human-to-human transmission. Currently, there are no approved treatments or vaccines for NiV infection in humans or animals. Consequently, there is a pressing need for a highly sensitive, precise, and visually detectable assay to enable early intervention and mitigate the transmission of NiV infection. Here, we report a single-copy sensitive, field-deployable, one-pot visual reverse transcription-recombinase polymerase amplification (RT-RPA)-clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR associate system (Cas)12 for the detection of NiV. The assay works by targeting the N gene of NiV, and the results are directly visible to the naked eye. The assay has demonstrated the ability to detect as few as 5.5 copies/μl of positive plasmids or 5.5 × 10¹ copies/μl of RNA transcripts when reacted at constant temperature for 40 min. It showed high specificity for NiV and had no cross-reaction with other pathogens, including rabies virus (RABV), Japanese encephalitis virus (JEV), herpes simplex virus type 1 (HSV-1), Hendra virus (HeV), and Streptococcus suis (S. suis), that can cause clinical symptoms similar to those of NiV infection. Moreover, this assay had a 100% coincidence rate with the reverse transcription quantitative polymerase chain reaction (RT-qPCR) method recommended by the World Organization for Animal Health (WOAH) for the detection of simulated clinical samples, indicating that it has great potential as an ultrasensitive, simple, and portable novel assay for the onsite diagnosis of NiV infection.

Rabbit hemorrhagic disease is a highly contagious and acute fatal disease caused by rabbit hemorrhagic disease virus (RHDV). The first outbreak of RHDV2 in 2020 has posed a serious threat to the rabbit breeding industry in China. An effective and specific detection strategy for RHDV GI.1 (RHDV1) and GI.2 (RHDV2) is urgently needed. In this study, we established a reverse transcription loop-mediated isothermal amplification (RT-LAMP)-CRISPR/Cas12a-based dual readout portable detection platform. The platform showed excellent specificity to identify RHDV1 and RHDV2 strains and no cross-reaction with other prevalent pathogens of rabbit. The detection limit for RHDV1 and RHDV2 by RT-LAMP-CRISPR/Cas12a could reach 10 copies/μl of the VP60 gene per reaction. Furthermore, 74 clinical samples were detected for both RHDV1 and RHDV2. RT-LAMP-CRISPR/Cas12a-based dual readout portable detection platform showed 25.68% (19/74) RHDV1-positive samples, 43.24% (32/74) RHDV2-positive samples, and 8.11% (6/74) RHDV1/RHDV2 double positive samples, respectively. The coincidence rates of detection RHDV1 and RHDV2 between RT-LAMP-CRISPR/Cas12a and quantitative real-time-polymerase chain reaction (qPCR) were both 97.30%. RT-LAMP-CRISPR/Cas12a showed higher sensitivity and detection rate compared with qPCR. Moreover, the results were visible to the naked eye within 1.5 h combined with lateral flow strips (LFSs) and visual fluorescence. The RT-LAMP-CRISPR/Cas12a portable platform has the advantages of high sensitivity, specificity, fast, low equipment requirements, which can be used in clinical practice in rural areas and resource-limited settings.

To date, seven novel parvoviruses have been identified in pigs and designated as porcine parvovirus 2–7 (PPV2–7). The presence of these emerging viruses has been reported in several countries around the world, although their pathogenic role and clinical and economical relevance are largely unknown. Here, we report the estimated prevalence and genetic diversity of novel PPV2–7 in Hungarian pig herds and the detection of these viruses in two Slovakian pig farms. For the comparative prevalence estimation, 2505 serum samples from different age groups, 218 oral fluid samples, and 111 processing fluid samples were collected from 26 large-scale Hungarian farms according to a systematic, cross-sectional sampling protocol. All samples were tested by real-time quantitative polymerase chain reaction (qPCR), and the presence of at least one PPV was detected in 24 of the 26 (92%) Hungarian and both Slovakian farms, suggesting high levels of subclinical circulation in most herds. The estimated PPV2–7 prevalence in Hungary varied from 50% to 89%, with PPV4 being the least and PPV2 being the most prevalent virus. The highest detection rates were observed in oral fluid samples, indicating that this sample type is most suitable for screening PPVs, but all viruses were also detected in serum samples and processing fluids. All novel PPVs were most frequently detected in the serum samples of weaned pigs and fatteners, with slightly higher viral burden in the younger age groups. These results may suggest an age-related susceptibility, which could play a significant role in the epidemiology of these viruses, impacting herd health and productivity.

Foot-and-mouth disease (FMD), which is endemic in 77% of countries globally, is a major threat to the global livestock industry. Knowledge of the reproductive number at the population level (i.e., farm level, herd level, or above) for FMD is important to estimate the magnitude of epidemics and design and implement effective control methods. Different methods, based on disparate assumptions and limitations, have been used interchangeably to compute and report reproductive numbers at the population level without a formal comparison between them. This study compares the results obtained when using alternative methods to compute between populations (Rbp) for FMD using one single dataset collected over 10 years (2007–2017) at the commune-level swine farms in Vietnam. Seven spatial–temporal clusters were identified in the country, and the value of Rbp was computed on each of them using different analytical approaches, namely, epidemic doubling time, nearest neighbor, time-dependent reproductive number (TDR), sequential Bayesian (SB), and birth–death skyline (BDSKY) analysis in Bayesian evolutionary analysis by sampling trees 2 (BEAST2). Estimated Rbp values were relatively similar across methods ranging from 1.25 to 1.61. For the first time, the results here provide a comparison of different methods used to compute Rbp for FMD. Despite differences in assumptions and limitations, results suggest that different methods produce relatively similar outputs. Additionally, the results here provide foundational knowledge to support the evaluation and control of FMD epidemics in a population.

In Africa, livestock production plays a crucial role for sustainable food security and economic growth. However, the development of this sector has been delayed by livestock diseases, one of the main constraints, which can cause important production and economic losses. To overcome these constraints, farmers extensively use antimicrobials, which in turn can lead to antimicrobial resistance (AMR), one of the main threats to global health and food security. Biosecurity has been identified as a key strategy to reduce livestock diseases. Therefore, the current systematic review and meta-analysis, conducted according to the Cochrane guideline, aimed at determining the efficacy of biosecurity in preventing and controlling infectious diseases in livestock farms in Africa. Of the 1408 records retrieved from five different databases, only 16 met the inclusion criteria. These studies were conducted in Egypt (31.2%), Nigeria (31.2%), Uganda (18.8%), Ethiopia (12.5%) and Tunisia (6.3%) and concerned poultry (62.4%), pigs (18.8%) and cattle (18.8%). Investigations focused mainly on avian influenza (AI) (15.0%) and coccidiosis (10.0%) in poultry and African swine fever (ASF) (10.0%) in pigs. In poultry farms, the results of the pairwise meta-analysis showed that biosecurity measures related to visitors and farmworkers could be effective at reducing the risk of introduction and spread of AI viruses (odds ratio [OR] = 0.48; 95% confidence interval [CI] 0.28–0.82). Moreover, inadequate biosecurity seemed to be a factor promoting coccidiosis (OR = 4.20; 95% CI 2.4–7.4) and AI (OR = 1.74; 95% CI 1.23–2.48). Prevention of ASF was significantly associated with the application of biosecurity measures related to animals’ transport, removal of carcasses and manure (OR = 0.33; 95% CI 0.12–0.88). Despite their importance, these findings cannot be translated to the entire African continent, since no studies were available for more than 90% of its countries. More research should be carried out to fill in the gaps identified by this review.

Feline coronavirus (FCoV) is an enveloped, positive-sense RNA virus, which is widespread among feline populations, and can cause a fatal serious disease called feline infectious peritonitis (FIP). According to the differences of antigen and genetic composition, FCoV consists of two genotypes, FCoV I and FCoV II. In this study, we have isolated and identified a FCoV I strain named HL2019. Based on the complete genome of HL2019, phylogenetic analysis showed that HL2019 strain formed in the cluster FCoV I which is more closed to human coronavirus 229E (HCoV 229E) and HCoV NL63, while the FCoV I stains is distantly related to FCoV II strains. Analyzing with RDP4 and Simplot software showed that the virus HL2019 is recombinant by the FCoV I China/ZJU1709 and FCoV I Netherlands/UU16 strains. Furthermore, the pathogenicity of HL2019 was evaluated in 9–12-month-old cats. Two of three challenged cats developed serious clinical signs and died at 28-day postchallenge (dpc). Real-time polymerase chain reaction (PCR) analysis showed that HL2019 has broad tissue tropism, especially in the duodenum with viral load up to 10⁴ copies/mg. In summary, our data show that we have successfully isolated a strain of FCoV I named HL2019 that is highly pathogenic to cats.

A high pathogenicity avian influenza (HPAI) subtype H7N6 virus emerged in South African poultry in 2023 and later spread to Mozambique, the first documented emergence of H7 HPAI in the African continent. A total of 6.82 million birds succumbed to the disease or were culled, representing about 20% of the South African egg-laying flock and almost 30% of the broiler breeder flock. The complete genomes of 68 outbreak viruses were sequenced and analyzed, tracing the phylogenetic origins of the ancestral H7N6 virus to a reassortment of various subtypes that circulated in southern African wild birds. Molecular clock analysis determined that the virus emerged in the first week of May 2023, probably in a smallholder chicken flock, before spreading to commercial farms, where the disease was first reported in early June. The multibasic hemagglutinin protein cleavage site (HA0) was derived from a nonhomologous recombination event with chicken 28S ribosomal ribonucleic acid (RNA). Few genetic markers associated with an increased risk to humans were present in the translated viral proteins. The intravenous pathogenicity index (IVPI) value of the index case isolate was 1.67, reflecting that 50% of the specific pathogen-free chickens died within 4 days of infection. Surviving birds showing mostly mild clinical signs and recovered by day 10 postinfection. Prior to death, chickens shed the virus primarily through the respiratory route, with lower amounts shed from the cloaca, but in the survivors, the virus was still being shed from the cloaca on day 10. Fomites were the likely source of disease spread between farms, and the amount of H7N6 HPAI virus per gram of feces was calculated at ~383,193 (5.58 log10) egg infectious dose 50 (EID50) equivalents, chicken feather follicles contained on average 739,712.43 (5.87 log10) EID50 equivalents, and 20 µg of feather dust contained 14,976.96 (4.175 log10) EID50 equivalents.

Wild rodents and their ectoparasites are known reservoirs for various zoonotic pathogens, highlighting the need for detailed studies into their roles in disease transmission. Our research investigated the spatial distribution of rodents and their ectoparasites to better understand the epidemiology of vector-borne zoonotic diseases (VBZDs), including severe fever with thrombocytopenia syndrome (SFTS), Lyme disease, Q fever, and scrub typhus. We analyzed samples from 540 rodents and 6785 mites, detecting the presence of Borrelia spp., the causative agent of Lyme disease, in 0.9% of rodents and SFTS virus (SFTSV) in 1.0%. In mites, Borrelia spp. and Orientia tsutsugamushi, the bacteria causing scrub typhus, were detected in 0.3% of samples each. Phylogenetic analysis identified the SFTSV sequence as type B3, the Borrelia spp. sequence as B. afzelii, and the O. tsutsugamushi sequence as Karp-related. Notably, SFTSV was detected for the first time in mites in South Korea, and B. afzelii was found in mites for the first time globally. These findings emphasize the critical need for continuous analysis of VBZDs to anticipate future trends and develop a comprehensive monitoring system. Further research into the rodent and mite populations in South Korea is essential to fully assess the potential risks of VBZDs.

Coronaviruses (CoVs) belonging to the Gamma-CoV and Delta-CoV genera are widespread in poultry and wildfowl. Migratory birds, particularly duck species, serve as hosts for CoVs and play a pivotal role in transmitting the viruses to other species, including mammals. Despite the potential risks to animals and humans, there remains a narrow knowledge of the genetic and epidemiological properties of CoVs in wild birds. The current research aimed to detect and characterize CoVs present in migratory duck species (Anas acuta, Anas platyrhynchos, and Anas poecilorhyncha) from South Korea. Employing two rounds of pan-CoV real-time reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR (nPCR) assays amplifying the conserved RNA-dependent RNA polymerase (RdRp) portion common to all known CoVs, we screened 2120 duck fecal samples collected during 2022–2023. The results indicated the presence of CoVs in 4.2% (91/2120) of samples from migratory ducks. Nucleotide sequencing of the RdRp gene revealed that all identified CoVs were clustered within the Gamma-CoV genus. Further phylogenetic analysis suggested that South Korean gamma-CoVs belong to the Igacovirus subgenus and share similarities with those found worldwide, highlighting the critical role of migratory ducks in introducing and exporting avian CoVs. We discovered two clade VII igacovirus strains in wild ducks closely related to those in pigeons, implying potential cross infection between these avian species. Overall, our study underscores the importance of active surveillance and monitoring of avian CoVs in wild birds as a preemptive response against the forthcoming emergence of new CoV species that can threaten both animal and human health.

Bovine viral diarrhea (BVD) represents a serious threat to the cattle sector in Chile, indicating the need for a regionally defined control program. Ex-ante evaluations of program options using simulation modeling have proven to be a successful approach in providing decision-makers with relevant supporting insights in that respect. Given the complexity of bovine viral diarrhea virus (BVDV) infection dynamics, simulation of BVD spread in a metapopulation requires detailed consideration of both within and between herd transmission dynamics. The aims of the study are (i) to investigate the dynamics of BVDV transmission in cattle herds in southern Chile by linking a within-herd transmission model (WHM) that accounts for the BVDV’s unique characteristics with a between-herd model (BHM) that meets the demands for further regional control strategy evaluation; (ii) to suggest and discuss criteria for evaluation of the model approach and plausibility for later research and for support decision-making. This resulted in bringing forth a modeling rationale for complex disease spread simulation in metapopulations. BHM simulations under this approach show outcomes that agree with BVDV’s known situation in Chile; dairy herds prevalence at endemic equilibrium reaches and maintains 75%, which agrees with estimations of BVDV active infection in dairy herds in southern Chile (77%). For the entire herd population, the infection always reaches endemic levels with a large proportion of infected herds (median = 60%), where herd prevalence was higher in the dairy herd class than in the remaining categories. Transmission probability variation affects the new infections picked, prevalence at endemic levels, and the velocity in which the infection spreads between herds. The fact that the presented approach was able to model a complex infection dynamic such BVDV, with sufficient confidence, provides evidence that this approach can be used to explore mitigation strategies to control BVDV in southern Chilean herds.

This study is the first report in Italy on the molecular characterization by whole-genome sequencing (WGS) analysis of a Streptococcus suis strain isolated from a human case of meningitis in Italy. The characterized S. suis strain was classified as a serotype 2 (SS2), multilocus sequence typing (MLST) sequence type ST1. The strain exhibited the presence of several virulence genes and resistance to penicillin, tetracycline and macrolide–lincosamide–streptogramin. Finally, we found a frameshift mutation in the gene mrp determining the translation of two truncated forms of the corresponding muramidase-release protein. These results highlight the importance of complete genomic data to understand the pathogenesis and epidemiology of this bacterium, capable to pose serious risks to human health.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a significant pathogen in the worldwide swine industry. The virus shows high genetic variation coupled with a broad range of virulence in pigs. Although multiple lineages of the virus have been prevalent throughout in Korea, the characteristics of lineage-wise pathogenicity are largely unknown. Therefore, this study was designed to analyze and compare the pathogenicity of 11 representative Korean PRRSV-2 isolates selected from PRRSV-2 lineages circulating in Korea, NADC30-like, VR2332-like, and three nation-specific lineages (lineage KOR A (LKA), lineage KOR B (LKB), and lineage KOR C (LKC)), which have been continuously prevalent in the nation. Eleven groups of pigs were experimentally infected with one Korean PRRSV-2 isolate through four consecutive animal experiments. Body weight and body temperature were recorded during each 4-week challenge experiment period, and virological, serological, and histopathological tests were performed on the collected samples. The data from the animal experiments were integrated into two indicators—excretion and clinical signs—through correlation and principal component analysis (PCA). Meta-analysis was used to compare PRRSV-2 isolates using each indicator. Based on these analyses, while L1C viruses used in this study (JB15-N-P31-GB and JB15-N-PJ73-GN, similar to NADC30-like strains) exhibited low or moderate levels of excretion and clinical signs, lineage 5 (L5) or modified live vaccine (MLV)-variant strains exhibited high levels of excretion compared to other PRRSV-2 isolates. However, the L5 variants all caused mild clinical signs, except for JB15-N-PJ4-GN, which showed the 4th highest clinical sign indicator. Among the Korean lineages (LKA, LKB, and LKC), two LKB strains (GGYC45 and JB15-N-PJ10-GN) were the most virulent as they showed the highest mortality after the challenge. On the other hand, the LKA and LKC viruses displayed lower excretion indicators than L5 strains, but they had higher-ranked clinical sign indicators than low-virulence L5 MLV variants. In conclusion, PRRSV prevalent in Korea has diverse excretion and clinical characteristics, and certain lineage is highly pathogenic. These results will offer useful insights to prevent spread of PRRSV and improve the efficacy of vaccines in the future.

Respiratory infections are a leading cause of death in developing countries. Infants and young children are especially susceptible to disease because they lack immunity, whereas adults who have acquired immunity can be infected asymptomatically. Great ape species, all of which are endangered, are similarly susceptible to respiratory illnesses caused by human respiratory pathogens. We obtained 432 nasopharyngeal swab samples (127 from adults and 305 from children) in a cross-sectional study that took place between February and October 2022 at four sites in Western Uganda (Budongo Central Forest Reserve, Bulindi Town Council, Bwindi Impenetrable National Park, and Kibale National Park) where the participants live in communities where interaction with apes is frequent. Prior research at Kibale has shown that locally circulating human respiratory pathogens have led to multiple lethal outbreaks in wild eastern chimpanzees (Pan troglodytes schweinfurthii). We used a multiplex PCR panel to characterize respiratory pathogens, with the goal of assessing whether respiratory illnesses in the chimpanzees of Budongo and Bulindi and the mountain gorillas (Gorilla beringei beringei) of Bwindi might have originated in local children and been introduced to the apes via asymptomatic adult carriers. The prevalence of respiratory pathogens was twice as high in Bwindi (44.0%) as it was in Budongo (24.0%) and Bulindi (20.8%), while the prevalence was intermediate at Kibale (34.4%). Rates of respiratory pathogen detection were higher but statistically indistinguishable in children compared to adults at Budongo and Bulindi, and children were 15.0 times more likely than adults to have positive detections at Kibale. At Bwindi, however, the pattern was reversed, with adults 2.6 times more likely than children to have positive detections. Rhinovirus, metapneumovirus, human parainfluenza virus 3, respiratory syncytial virus, and coronavirus OC43, all of which have been identified as causative agents of respiratory disease outbreaks in great ape populations across sub-Saharan Africa, accounted for three quarters (73.6%) of detected pathogens. Our data support the idea that human respiratory pathogens that can infect great apes occur at high frequencies in human populations in Western Uganda that live close to and interact with wild apes that have suffered from lethal outbreaks caused by these same pathogens. Reducing respiratory infections in local children, thereby reducing both carriage of those infections into the forest by people and ape exposure to these pathogens when they enter human spaces, should decrease the risk of respiratory disease outbreaks in apes.

The highly pathogenic avian influenza 2.3.4.4b H5 viruses have been a cause for concern recently, as they have been responsible for continuous outbreaks since 2021. In China, the H5N6 subtype has been predominantly circulating in domestic poultry but has rarely been detected in wild birds over the past 3 years. In December 2023, novel reassortant 2.3.4.4b H5N6 viruses were resurgent in wild birds and domestic ducks in Eastern Asia. The viruses were reassorted with those of currently prevalent 2.3.4.4b H5N1 viruses of wild bird origin worldwide, as well as the H5N6 viruses that caused human infections in 2022 and low pathogenic avian influenza viruses, such as the H9N2 virus, which also contributed internal gene to the novel H5N6 viruses. Based on the phylogenetic analyses, we inferred that this recombination process occurred in migratory breeding sites in early 2023. Given the rapid transmission and high mutation capacity of currently circulating H5N1 viruses, as well as the strong pathogenicity of H5N6 viruses to humans, the novel recombinant viruses may continue to evolve and pose new threats to human health. Therefore, continuous surveillance of H5N6 viruses in wild birds and domestic poultry should be strengthened.

Porcine reproductive and respiratory syndrome virus (PRRSV), encompassing PRRSV-1 and PRRSV-2, significantly impacts the global pig industry by causing reproductive disorders and respiratory difficulties. In this paper, we isolated a novel PRRSV-1 strain, named SCPJ2023, from weaned piglets in Sichuan. Utilizing primary macrophages, we isolated SCPJ2023 and performed complete genome sequencing through metagenomic analysis. Phylogenetic analysis classified SCPJ2023 as pan-European subtype 1. SCPJ2023 showed a 95.3% similarity to SC2020-1. Amino acid analysis identified differences in Nsp2, GP3, and GP4 between SCPJ2023 and other representative strains. In vivo challenge experiments demonstrated that SCPJ2023 induced clinical symptoms in piglets, including coughing, fever, reduced appetite, and depression. Pathological examinations revealed hemorrhage and congestion, increased inflammatory cells, thickening of the alveolar wall, and collapse of the alveolar cavity in SCPJ2023-infected piglets. Altogether, our study identified a novel pathogenic isolate of PRRSV-1, expanding the newly named SC2020-1-like subgroup by identifying additional strains beyond the initial SC2020-1 isolate.

Anthropogenic activities in the environment affect the ecosystem and can play an important role in selecting and spreading antibiotic-resistant bacteria (ARB) and genes (ARGs). The dairy farm environment may serve as a hotspot and reservoir for exchanging and spreading ARGs, but studies are scarce. Here, we investigated and characterized the extended-spectrum β-lactamase producing Escherichia coli strains recovered from the dairy farm environment co-harboring blaOXA−1, catB3, and arr-3 genes. The isolates were identified and characterized by PCR, antimicrobial susceptibility testing, conjugation assay, whole genome sequencing (WGS), and multiple bioinformatics tools. Seven E. coli strains co-harboring blaOXA−1, catB3, and arr-3 genes were identified which belonged to distinct sequence types (STs) and carried diverse plasmid replicon types. The conjugation assay revealed a successful transfer of blaOXA−1, catB3, and arr-3 genes into the recipient E. coli J53 with a co-conjugation frequency ranging from (2.25 ± 0.3) × 10⁻⁴ to (3.85 ± 0.3) × 10⁻³. Bioinformatics analysis of WGS revealed the diversity of acquired ARGs, conferring resistance to aminoglycosides, beta-lactams, quinolones, tetracyclines, macrolides, trimethoprim–sulfamethoxazole, phosphonic, phenicol, and rifamycin. The genetic environment analysis showed that aac(6′)-Ib-cr-blaOXA−1-catB3-arr-3-qacE1-sul1 was the common genetic backbone among the seven E. coli strains. Among the mobile genetic elements, insertion sequences were the predominant elements as compared to transposons. The phylogenetic analysis demonstrated a close relationship between the E. coli of this study and other strains of human–animal-environment origin retrieved from the NCBI database. This study presented the whole genome-based characterization of E. coli strains carrying the blaOXA−1-catB3-arr-3 genes. It provided evidence that the dairy environment may harbor a variety of ARGs and act as a potential reservoir for their spread in the ecosystem. The results recommend the routine surveillance of ARGs carrying bacteria in dairy environments and the need for additional studies to understand the dissemination mechanism within One Health perspective to prevent their further spread.

The growing prevalence of commensal antibiotic resistant Escherichia coli poses a significant concern for the global spread of antibiotic resistance. Stool samples (n = 35) from a seminomadic indigenous community in Malaysia, the Jehai, were screened for multidrug-resistant bacteria, specifically the extended-spectrum β-lactamase (ESBL) producers. Subsequently, whole-genome sequencing was used to provide genomic insights into eight ESBL-producing E. coli that colonised eight individuals. The ESBL E. coli isolates carry resistance genes from various antibiotic classes such as the β-lactams (blaTEM, blaCTX-M−15 and blaCTX-M−55), quinolones (gyrA, qnrS and qnrS1) and aminoglycosides (aph(3′)-Ia, aph(6)-Id and aac(3)-IId). Three concerning convergence of ESBL, colistin and metal resistance determinants, with three plasmids from H-type lineage harbouring blaCTX-M and mcr-1.1 genes were identified. Using the Oxford Nanopore Technology (ONT) Native Barcoding Kit (SQK-NBD114.24) in conjunction with the R10.4.1 flow cell, which achieved an average read accuracy (Q > 10) of 99.84%, we further characterised the mcr-1.1-bearing plasmids, ranging in size from 25 to 28 kb, from three strains of E. coli. This report represents the first whole genome analysis of multidrug-resistant bacteria, specifically those resistant to colistin, found within the indigenous population in Malaysia. It strongly indicates that the pertinent issue of colistin resistance in the country is far more significant than previously estimated.

We aimed to understand the temporal dynamics of SARS-CoV-2 reinfection risk and assess the impact of inactivated vaccination on the occurrence of reinfection. We investigated the reinfection risk of SARS-CoV-2 from November 1, 2022, to February 12, 2023, when China rapidly lifted the zero-COVID policy. The study subjects were those who were first infected during the zero-COVID period between January 1, 2020, and October 31, 2022, in Dalian city, China. Among the 1961 previous infections, 126 (6.4%, 95% CI: 5.4, 7.5) were reinfected. The risk of reinfection increased over time since initial infection. Compared with those who did not receive or received one dose of inactivated vaccine, receiving two or three doses was associated with additional protection against reinfection among individuals who were infected with pre-Omicron more than a year earlier, with the OR ranged from 0.33 (95% CI: 0.03, 1.83) to 0.91 (95% CI: 0.22, 3.27). In contrast, no protective effect from two or three doses of vaccines against reinfection was observed among those who were first infected with Omicron variants within a year. Primary or booster vaccination contributed to limited protection against reinfection or symptomatic reinfection among individuals infected with Omicron SARS-CoV-2 within a year. However, a booster dose after 1 year of natural infection may provide additional protection against reinfection.