To gain insight into the molecular epidemiology and possible mechanisms of genetic variation of porcine reproductive and respiratory syndrome (PRRS) in Yunnan Province of China, the ORF5 gene of 32 PRRSV isolates from clinical samples collected from 2007 to 2009 were sequenced and analyzed. Nucleotide and amino acid analyses were carried out on 32 isolates and representative strains of the North American genotype, European genotype and two representative Chinese isolates. Results revealed that these isolates share 86.9–99.0% nucleotide and 87.5–98.0% amino acid identity with VR-2332 the prototypical North American PRRSV, 61.7–62.9% and 54.3–57.8% with Lelystad virus (LV) the representative strain of European genotype, 91.2–95.4% and 90.0–94.5% with CH-1a that was isolated in mainland China in 1996, 88.1–99.3% and 85.5–99.0% with JX-A1 the representative strain of High pathogenic PRRSV in China, and 86.2–99.8% and 85.5–100.0% between isolated strains of different years, respectively. Phylogenetic analysis revealed that all 32 PRRSV isolates belonged to the North American genotype and were further divided into two different subgenotypes. Subgenotype 1 comprised twenty two Yunnan isolates which divided into two branches. Subgenotype 2 comprised ten isolates which closely related to the RespPRRS vaccine and its parent strain VR-2332. The functional domains of GP5 such as the signal peptide, ectodomain, transmembrane regions and endodomain were identified and some motifs in GP5 with known functions, such as primary neutralizing epitope (PNE) and decoy epitope were also further analyzed. Our study shown the great genetic diversity of PRRSV in southwest China, rendering the guide for control and prevention of this disease.

Porcine reproductive and respiratory syndrome (PRRS) is caused by porcine reproductive and respiratory syndrome virus (PRRSV) and is an economically important disease in swine-producing areas. The objective of this study was to screen for effective antisense oligonucleotides (AS-ONs) which could inhibit PRRSV replication in MARC-145 cells and in pulmonary alveolar macrophages (PAM). Nine short AS-ON sequences against the well-conserved regions of PRRSV (5′-UTR, NSP9, ORF5 and ORF7) were selected. When MARC-145 cells or PAM were infected with PRRSV followed by transfection with AS-ONs, four AS-ON sequences targeting 5′-UTR, ORF5 or NSP9 were found to be the most effective oligonucleotides in decreasing the cytopathic effect (CPE) induced by PRRSV infection. Quantitative PCR and indirect immunofluorescence staining confirmed that ORF7 levels were significantly reduced both at RNA and protein levels. The PRRSV titration data furthermore indicated that transfection with AS-ON YN8 could reduce the PRRSV titer by 1000-fold compared with controls. The results presented here indicate that DNA-based antisense oligonucleotides can effectively inhibit PRRSV replication in MARC-145 cells and in PAM. Furthermore, comparing with the reported hit rates (approximately 10-30 %), we achieved a higher success rate (44 %). The strategy we took to design the antisense sequences might be applied to select AS-ONs that more efficiently reduce the expression of target genes.

Hepatitis E is a disease of major public-health concern mainly in developing countries. Although molecular and sero-epidemiological investigations of HEV have been performed in many provinces in China, the epidemiological data from Yunnan Province are limited and genotypes are not be fully characterized. In this study the prevalence and characteristics of hepatitis E virus (HEV) detected in pigs from Yunnan province, China was evaluated. A total of 13 out of 187 pig fecal samples collected in 2011 revealed HEV positive results; likewise, 7 out of 69 samples collected in 2012 exhibited positive results. These findings indicated a total prevalence of 7.8% (20/256). Phylogenetic and molecular evolutionary analysis results revealed that nine strains were found in the samples obtained in 2011, in which 87.1% to 99.4% nucleotide sequence identity was shared among these strains; and 77.0% to 81.9%, 52.2% to 53.6%, 77.0% to 88.2% and 77.9% to 96.8% nucleotide sequence identities were shared with strains representing genotypes 1, 2, 3, and 4. Five strains were detected in the samples obtained in 2012, in which 94.2% to 99.3% nucleotide sequence identity was shared among the strains, and 81.0% to 82.5%, 81.8% to 83.2%, 81.0% to 92.7% and 81.0% to 97.8% nucleotide sequence identities were shared with strains representing the genotypes 1, 2, 3, and 4. Analysis of fourteen detected HEV strains revealed that three of them were subtype 4d, two were subtype 4b; the nine remaining isolated strains were subtype 4 h. These results indicated that the prevalence of HEV in the swine herds of Yunnan was quite high, additional public-health concerns should focus on pork safety.

Background: Mobile antimicrobial resistance genes (ARGs) on plasmids or other elements enable Salmonella Typhimurium to spread resistance across hosts and environments. The emergence of multi-drug resistance (MDR) Salmonella Typhimurium has raised global concern, yet little is reported about these mobile elements from the Thailand pork supply chain, where this risk of transfer to humans remains largely uncharacterized. Methods: Between March 2023 and February 2024, 25 S. Typhimurium isolates were collected from pig carcasses in slaughterhouses and pork swabs from retail markets in northeastern Thailand. Nine representative isolates, sampled across three seasons, were subjected to Illumina whole-genome sequencing. Assemblies were analyzed for sequence types, phylogenetic relationships, antimicrobial resistance (AMR) determinants, plasmid replicons and mobilization features, functional annotation based on COG (Clusters of Orthologous Groups of proteins) classification, and comparative genomics against a reference strain. Results: Genome assemblies ranged from 4.76 to 5.00 Mb with consistent GC (guanine-cytosine) content (52.0–52.2%). Phylogenetic analysis revealed three sequence types: ST34 (77.8%), ST19, and ST1543. ST34 isolates displayed the broadest AMR gene repertoires, carrying tetracycline (tetA/tetB), sulfonamide (sul1/sul2/sul3), aminoglycoside (aadA, aph(6)-Id, aph(3″)-Ib), phenicol (floR, catA1), and β-lactam (bla_TEM-1B) genes, whereas non-ST34 isolates harbored fewer determinants. ARGs frequently co-localized with IncQ1 and Col-type plasmid replicons, MOB_H/MobA relaxases (enzymes that initiate plasmid transfer), and conjugation modules (type IV secretion and coupling proteins), often alongside virulence loci and metal resistance operons. Functional annotation showed highly conserved metabolic and housekeeping functions, while comparative genomics confirmed >90% core genome conservation, with variability concentrated in genomic islands encoding hypothetical proteins. These genomic patterns were inferred from a limited WGS dataset (nine isolates) and should therefore be considered exploratory and require confirmation in larger collections. Conclusions: Multi-drug resistant ST34 Salmonella Typhimurium predominated in the northeastern Thailand pork supply chain, with diverse resistance genes carried on IncQ1/Col-type plasmids linked to MOB_H relaxases and conjugation modules. The stability of these mobilizable elements underscores their role in sustaining MDR traits and highlights the risk of foodborne AMR transmission, reinforcing the need for continuous genomic surveillance under a One Health framework.

Bovine Kobuvirus (BKV) is an emerging pathogen associated with diarrhea in cattle. Limited reports on its prevalence and genetic characteristics are available. To determine the epidemiology and genetic evolution of BKV strains circulating in Yunnan Province, China, 204 diarrheal samples were collected from cattle farms across five regions for screening for BKV infection. RT-PCR analysis identified 40 BKV-positive samples, yielding an infection rate of 19.6%. Positive samples were inoculated into Vero cells for continuous passage, followed by molecular biology, immunofluorescence, and electron microscopy identification. Two BKV strains, BKV YN-1 2023 and YN-2 2023, were isolated. Whole-genome sequencing revealed genome lengths of 8289 bp and 8291 bp (GenBank No. PV410179 and PV410180), respectively. Phylogenetic analysis demonstrated that both strains belong to genotype B, the dominant genotype circulating in China, and are closely related to the previously reported Chinese strain BKV 13/2021. The genetic similarity of two BKV isolates was analyzed. Genome-wide nucleotide identities ranged from 39.9 to 93.9%, with the highest similarity to BKV13 2021 CHN. ORF analysis showed nucleotide and amino acid similarities of 48.7–93.9% and 29.3–98.5%, respectively. Compared to the BKV13 2021 CHN, both isolates exhibited high conservation in VP0, VP3, and nonstructural proteins (97.8–100%), while the L protein had the lowest similarity (94.7–95.2%). The 5′ UTR showed lower conservation than the 3′ UTR, suggesting regulatory variations. These findings reveal that the circulating BKV strains in Yunnan belong to the globally prevalent genotype B and are widely distributed. This study provides valuable insights into the molecular epidemiology, genetic diversity and evolutionary dynamics of BKV, offering an important reference for developing diagnostics, vaccines and further studies on its pathogenesis.

Lumpy skin disease (LSD) poses a significant transboundary threat to livestock health and productivity, especially in regions where vector-borne transmission is a major driver of spread. Environmental and climatic factors are recognized as critical determinants of LSD transmission dynamics. This study aimed to model the environmental suitability for LSD across Thailand using a maximum-entropy approach. Outbreak data from 2021 to 2023 were analyzed alongside bioclimatic variables, land cover, normalized difference vegetation index (NDVI), and cattle population density. The model produced an area under the curve (AUC) value of 0.699 (~0.70), indicating moderate predictive performance. Based on variable contribution, land cover (65%), cattle density (25%), and NDVI (3%) were identified as the most influential predictors of environmental suitability for LSD. The resulting risk map identified central and northeastern Thailand as the most suitable regions for disease occurrence. These findings provide valuable insights to support risk-based surveillance, improve veterinary resource allocation, and enhance early warning systems for effective LSD prevention and control in Thailand.