In recent years, the prevalence of pseudorabies virus (PRV) has caused huge economic losses to the Chinese pig industry. Meanwhile, PRV infection in humans also sounded the alarm about its cross-species transmission from pigs to humans. To study the regional PRV epidemic, serological and epidemiological investigations of PRV in pig populations from Yunnan Province during 2017–2021 were performed. The results showed that 31.37% (6324/20,158, 95% CI 30.73–32.01) of serum samples were positive for PRV glycoprotein E (gE)-specific antibodies via enzyme-linked immunosorbent assay (ELISA). The risk factors, including the breeding scale and development stage, were significantly associated with PRV seroprevalence among pigs in Yunnan Province. Of the 416 tissue samples collected from PRV-suspected pigs in Yunnan Province, 43 (10.33%, 95% CI 7.41–13.26) samples were positive for PRV-gE nucleic acid in which 15 novel PRV strains from these PRV-positive samples were isolated, whose gC and gE sequences were analyzed. Phylogenetic analysis showed that all 15 isolates obtained in this study belonged to the genotype II. Additionally, the gC gene of one isolate (YuN-YL-2017) was genetically closer to variant PRV strains compared with others, while the gE gene was in the same clade with other classical PRV strains, indicating that this isolate might be a recombinant strain generated from the classical and variant strains. The results revealed the severe PRV epidemic in Yunnan Province and indicated that PRV variants are the major genotypes threatening the pig industry development.

Porcine epidemic diarrhea virus (PEDV) is a highly contagious pathogen responsible for devastating enteric disease and lethal watery diarrhea, leading to significant economic losses in the global swine industry. Understanding the epidemiology and genetic diversity of PEDV over the past decade is crucial for the effective prevention and treatment of porcine epidemic diarrhea. In this study, 1851 fecal samples were collected from pigs exhibiting diarrhea symptoms across 11 cities in Yunnan Province between 2013 and 2022. The prevalence of PEDV, along with other common swine diarrhea viruses, including porcine transmissible gastroenteritis virus (TGEV), porcine rotavirus (PoRV), porcine Sapporo virus (PoSaV), porcine stellate virus (PaStV), and porcine delta coronavirus (PDCoV) was assessed using a polymerase chain reaction (PCR) assay. The results revealed a total detection rate of 52.94% (980/1851) for the six viruses, with PEDV accounting for 25.93% (480/1851) of cases. Further analysis showed that weaned piglets were more susceptible to PEDV than fattening pigs, with the highest prevalence observed in spring (61.52%, 275/447) and the lowest in summer (12.68%, 97/765). Dual infections were also identified, with PEDV + PoSaV being the most common combination (2.81%, 52/1851), followed by PEDV + PoRV, with a detection rate of 1.67% (31/1851). Phylogenetic analysis of the PEDV S genes revealed that the 28 epidemic strains in Yunnan Province shared a nucleotide sequence homology from 91.4% to 98.4% and an amino acid sequence homology ranging from 85.6% to 99.3%. All strains were classified as GII variant strains. This study provides a comprehensive overview of the epidemiology of PEDV and its co-infection patterns with other common diarrhea-causing viruses in the swine herds of Yunnan Province over the past decade. These findings offer valuable insights for the development of effective prevention and control strategies to mitigate the impact of PEDV and other enteroviruses on the swine industry in Yunnan Province.

Enterovirus G (EV-G) is prevalent in pig populations worldwide, and a total of 20 genotypes (G1 to G20) have been confirmed. Recently, recombinant EV-Gs carrying the papain-like cysteine protease (PLCP) gene of porcine torovirus have been isolated or detected, while their pathogenicity is poorly understood. In this study, an EV-G17-PLCP strain, ‘EV-G/YN23/2022’, was isolated from the feces of pigs with diarrhea, and the virus replicated robustly in numerous cell lines. The isolate showed the highest complete genome nucleotide (87.5%) and polyprotein amino acid (96.6%) identity in relation to the G17 strain ‘IShi-Ya4’ (LC549655), and a possible recombination event was detected at the 708 and 3383 positions in the EV-G/YN23/2022 genome. EV-G/YN23/2022 was nonlethal to piglets, but mild diarrhea, transient fever, typical skin lesions, and weight gain deceleration were observed. The virus replicated efficiently in multiple organs, and the pathological lesions were mainly located in the small intestine. All the challenged piglets showed seroconversion for EV-G/YN23/2022 at 6 to 9 days post-inoculation (dpi), and the neutralization antibody peaked at 15 dpi. The mRNA expression levels of IL-6, IL-18, IFN-α, IFN-β, and ISG-15 in the peripheral blood mononuclear cells (PBMCs) were significantly up-regulated during viral infection. This is the first documentation of the isolation and pathogenicity evaluation of the EV-G17-PLCP strain in China. The results may advance our understanding of the evolution characteristics and pathogenesis of EV-G-PLCP.

In order to investigate the causes of swine diarrhea in Yunnan Province, this study was conducted to detect and monitor diarrhea viruses through regular sampling and reverse transcription polymerase chain reaction (RT-PCR). In October 2023, porcine sapelovirus (PSV) was detected in fecal specimens collected from diarrheal pigs in Honghe City, and three strains of PSV were successfully isolated by inoculating them into PK-15 cells; electron microscopy revealed virus particles with diameters of ~32 nm. Next-generation sequencing (NGS) revealed that the PSV isolate genomes ranged from 7480 to 7515 nucleotides in length. Homology analyses indicated that ML-15 and ML-16 showed the highest nucleotide and amino acid identities with the Asian PSV strains, ML-19 showed the highest sequence identities with the Zambia PSV strains, and the VP1 to VP4 genes of the three PSV isolates were in the hypervariable region. Phylogenetic analyses showed that the three PSVs isolated in this study all clustered together with Chinese PSV strains; furthermore, recombination analyses indicated that PSV-ML-19 might be a recombined strain and may have emerged through genetic recombination between the major putative parent strain PSV-21-V and the minor putative parent GER L00798-K11 14-02. This was the first reported instance of the isolation and phylogenetic analyses of the PSV strains in Yunnan Province, which enriched the understanding of Chinese PSV strains and indicated the need to prevent and control PSV; the mutation of the VP1 and 3D genes may also provide an important reference for the development of PSV vaccines.

Porcine Sapelovirus (PSV) is widely prevalent in pig herds throughout the world and induces diarrhea, encephalomyelitis, respiratory tract symptoms, and reproductive disorders. However, the epidemiological and genetic evolution characteristics of PSV remain unclear in Yunnan Province. In this study, 1622 fecal samples were collected from pig farms in Yunnan Province. PSV and its co-infection rates with other pathogens were detected; then, the PSV VP1 gene was amplified and sequenced; and the genetic evolution characteristics of the VP1 gene were analyzed. The overall infection rate of PSV in Yunnan Province was 36.50%, and the differences among regions were significant (p < 0.05). The positive rates among different seasons were significantly different (p < 0.01), ranging from 73.33% (autumn) to 19.00% (summer). The PSV positive rate in diarrhea samples (47.26%) was significantly higher (p < 0.001) than that of non-diarrhea samples (31.77%). The co-infection rates of PSV with porcine rotavirus (PoRV) and PSV with porcine epidemic diarrhea virus (PEDV) were 5.07% and 3.04%. A total of 36 VP1 sequences were obtained, and the average identity among the 36 sequences was 85.3%, which was higher than that with other reference strains. Phylogenetic analysis revealed that all 36 PSV strains belonged to the PSV-1 genotype. The VP1 gene was under strong negative selection pressure (average dN/dS = 0.0838); however, the 95th amino acid was under positive selection pressure. Our study revealed the epidemiological, co-infection, and genetic evolution characteristics of PSV in pig herds of Yunnan Province, providing more data for preventing and controlling diarrhea pathogens in pigs.

Pseudorabies virus (PRV), also called suid alphaherpesvirus 1, is a double-stranded linear DNA virus belonging to the subfamily Alphaherpsvirinae of the family Herpesviridae