Abstract Feline panleukopenia virus (FPV) is a highly contagious parvovirus that causes acute gastroenteritis, leukopenia, and high mortality in felids. Although domestic cats are commonly vaccinated, FPV continues to threaten captive and wild felids because of its environmental stability, rapid progression, and potential antigenic drift. In June 2022, a one-year-old captive female cougar ( Puma concolor ) at Hongshan Forest Zoo, Nanjing, China, was found dead following a peracute course without prodromal signs. The animal had previously received a trivalent feline vaccine containing feline panleukopenia, herpesvirus, and calicivirus antigens. Postmortem examination revealed perianal fecal staining, oral discharge, pulmonary congestion with peripheral emphysema, a darkened liver, and intestinal mucosal sloughing with petechiae. Histopathology revealed crypt epithelial necrosis, mucosal sloughing, and lymphoid depletion, which was consistent with parvoviral enteritis. Fecal and intestinal samples tested positive for FPV according to the lateral-flow assay and PCR. Viral replication was confirmed in CRFK cells via indirect immunofluorescence, and FPV antigen was shown to be localized to the intestinal crypt epithelium via immunohistochemistry. VP2 gene sequencing (1,753 bp) revealed that the isolate clustered with field strains from domestic cats in Jiangsu and Shanghai, which share near-complete nucleotide identity but differ from the Felocell vaccine strain in three amino acid substitutions (A91S, I232V, and L562V), two of which lie in antigenic loops and may affect antigenicity. No further FPV cases were detected during or after the 14-day observation period, reflecting successful containment through disinfection, relocation, feral cat control, and movement restrictions. This represents the first confirmed fatal FPV infection in a captive cougar in China and highlights the potential for local spillover from domestic reservoirs. The case underscores the need for continuous molecular surveillance, vaccination evaluation, and One Health–based biosecurity to protect susceptible wildlife populations at the human–domestic–wildlife interface.

Abstract Nipah virus disease, caused by Nipah virus (NiV), is a zoonotic infectious disease with an extremely high fatality rate. Owing to the absence of effective countermeasures, Nipah virus disease has caused substantial economic losses in the swine farming industry. Vaccines represent the most cost-effective approach for prevention and control. Herein, we describe the development of a recombinant pseudorabies virus (PRV)-vectored vaccine (designated PRV-HNX-ΔTK/gE-NiV-G), which expresses the codon-optimized full-length glycoprotein of Nipah virus (NiV-G). The biological characteristics of this strain, including plaque morphology, growth kinetics, and genetic stability, were evaluated in vitro. Furthermore, the recombinant virus exhibited favorable safety profiles in mice. Intramuscular administration of PRV-HNX-ΔTK/gE-NiV-G elicited high-titer neutralizing antibodies against NiV-G and a robust cellular immune response in BALB/c mice. Taken together, these findings indicate that PRV-HNX-ΔTK/gE-NiV-G has potential as a promising candidate for the development of bivalent vaccines that target both NiV and PRV infections.

Abstract Lumpy skin disease (LSD), a highly contagious viral infection caused by Lumpy Skin Disease Virus (LSDV), has caused severe economic losses. We analyzed three representative outbreaks in 2024, combining clinical, histopathological, serological, and molecular data to refine regional control strategies. According to our findings, affected cattle presented with generalized cutaneous nodules, fever and peripheral edema; one-third of in-contact animals remained clinically normal yet seroconverted within ten days. Antibody titers peaked at approximately 20 d post-onset and were accompanied by persistent viral DNA in saliva, ocular secretions and skin scabs. Histology revealed alveolar septal edema, hepatocellular ballooning, and glomerular necrosis, whereas biochemical profiling revealed hypalbuminemia, hyperglobulinemia, and elevated alanine aminotransferase in one patient. Immunohistochemistry revealed intense LSDV antigen in the spleen, alveolar septa of the lung, and throughout the dermis of the skin nodules, identifying these tissues as the most reliable diagnostic targets. The combined findings confirm active viral replication in cutaneous, respiratory and lymphoid tissues, highlight the occurrence of subclinical infection and underscore the value of paired serology and PCR for herd screening. Prompt isolation, vector control and vaccination with homologous capripox vaccines are recommended to curtail transmission and economic impact.

Abstract Sacbrood virus (SBV) is one of the most pathogenic honeybee viruses with host specificity and regional variation. The SBV strain infecting the Chinese honeybee ( Apis cerana ) is known as Chinese sacbrood virus (CSBV). The extensively used CSBV detection methods require professionals and expensive equipment; thus, they are unsuitable for rapid onsite CSBV detection. To achieve early and rapid detection of CSBV, we developed a lateral flow detection (LFD) strip method for CSBV detection via clustered regularly interspaced short palindromic repeats (CRISPR) and the Cas13a technique. On the basis of the conserved CSBV VP2 gene nucleotide region, we designed 3 recombinant enzyme-assisted amplification (RAA) primer pairs and prepared 3 corresponding crRNAs. We investigated key performance metrics, including the sensitivity, specificity, and accuracy of LFD strips. The results demonstrated that the LFD strip based on the optimal combination (primer 2+crRNA 2) presented the lowest detection limit (2.80×10 1 copies/μL), and this strip could complete CSBV detection within 1 h. Furthermore, this strip exhibited excellent detection specificity, with no cross-reactivity with four other honeybee viruses. A test of 100 clinical samples indicated the feasibility of the LFD method for CSBV detection. A comparison of various CSBV detection methods revealed that the CRISPR-Cas13a-based LFD method was more accurate, efficient, and sensitive than the other methods were, indicating great application prospects in onsite CSBV detection. Our developed method is highly important for preventing and controlling CSBV infection as well as maintaining honeybee health.

Abstract The rapid evolution of pathogen threats and pandemic risks has intensified the demand for adaptable vaccine platforms with rapid design-to-clinical timelines. The success of BNT162b2 and mRNA-1273 in controlling COVID-19 has led to unprecedented changes in vaccine development. Compared with traditional platforms, mRNA vaccines offer distinct advantages: avoiding genomic insertion risks of DNA vaccines; facilitating rapid, scalable production via in vitro transcription; and enabling new modular antigen design against evolving pathogens. However, the development of mRNA vaccines for infectious diseases still faces challenges, including structural stability, translation efficiency, and targeted delivery. This review presents cutting-edge strategies for designing antigens, optimizing mRNA structural elements, and optimizing delivery systems to target infectious pathogens. This study provides a detailed explanation of antigen optimization strategies for different types of mRNA vaccines and explores the potential applications of broad-spectrum and combination mRNA vaccines in human and animal pathogens. Finally, we address the current challenges and future prospects of mRNA vaccine technology, aiming to inform and accelerate the development of next-generation vaccines against emerging pathogens through the integration of these innovations.

Abstract Phosphorylation of the influenza A virus (IAV) nucleoprotein (NP) is crucial for the viral life cycle, as it encapsidates the viral genomic RNA. Here, we employed mass spectrometry to identify serine 69 (S69) as a novel phosphorylation site of NP in IAV. Sequence homology alignment revealed that NP S69 is highly conserved across various subtypes of IAV. The S69A mutation, which is designed to mimic constant dephosphorylation, had minimal effects on viral replication and pathogenicity in both cellular and murine models. In contrast, the S69E mutation, which mimics constant phosphorylation, was lethal. Mechanistically, the phosphorylation of NP S69 reduces the polymerase activity of viral ribonucleoprotein (vRNP) by impairing the interaction between NP and PB2. Additionally, NP S69 phosphorylation inhibited NP nuclear import by disrupting its interaction with importin-α3. These findings identify NP S69 phosphorylation as a critical regulatory mechanism in IAV replication, providing valuable insights into the role of NP phosphorylation in the life cycle of influenza virus.

Porcine epidemic diarrhea virus (PEDV) is a highly infectious intestinal coronavirus that poses a significant threat to the pig industry because of its strong virulence and propensity for mutation and recombination. PEDV is classified into three major genotypes based on the spike (S) gene sequence: G1 (classical), G2 (variant), and S-INDEL (characterized by nucleotide insertions/deletions in the S gene). Both the G1 and G2 genotypes have been frequently detected in China, whereas the S-INDEL strain has rarely been reported or isolated. In this study, we isolated a PEDV S-INDEL strain, designated EJS6, from a pig farm experiencing severe diarrhea in Jiangsu Province, China. Genetic evolution analysis revealed that the Chinese S-INDEL strains represented by EJS6 presented sequence differences from other S-INDEL strains isolated in Europe and the United States, forming a new branch within the S-INDEL genotype. Compared with the G2 strain EHuB4, EJS6 presented a lower viral titer, smaller plaque size, and reduced syncytium-forming ability in Vero cells. We also compared the pathogenicity of EJS6 and EHuB4 in 5-day-old suckling piglets. Both strains exhibited similar levels of fecal virus shedding in the infected piglets; however, the histopathological and mortality results indicated that the pathogenicity of the EJS6 strain was weaker than that of the EHuB4 strain. In summary, we successfully isolated the first Chinese PEDV S-INDEL strain and characterized its genetic evolution and pathogenicity, thereby enhancing our understanding of the prevalence of PEDV in China.

The contamination of pork during breeding can negatively impact both food quality and safety. Sodium dichloroisocyanurate (NaDCC), a chlorine-containing disinfectant, is widely used in animal environments. In this study, 16S rRNA sequencing was used to determine the bacterial communities in a pork farm. We also assessed the effectiveness of NaDCC disinfection by analyzing microbial diversity. The results revealed that the predominant genera in pork skin samples and environmental samples were Staphylococcus and Acinetobacter, respectively. Escherichia coli and Streptococcus equinus were present in all the samples, suggesting that NaDCC is not fully effective in preventing zoonotic pathogen contamination. The Chao1 and Shannon indices for sow skins increased after disinfection. No significant differences were observed in the microbiological composition of sow barn samples before and after disinfection (p > 0.1). Additionally, bacterial abundance in sow skin was strongly correlated with abundance on pen floors (r = 0.906, p < 0.05), indicating potential cross-contamination between these sample types. Conversely, the bacterial abundance in the floor samples was weakly correlated with that in the other samples, suggesting that NaDCC is effective as a decontaminant. This study provides valuable insights into microbial contamination on pork farms and underscores the importance of proper disinfection practices. This study also offers scientific recommendations for improving disinfection protocols.

Lumpy skin disease (LSD) poses a significant threat to cattle populations and the livelihoods of farmers in Thailand. This study uses path analysis to explore the interrelationships among the knowledge, attitudes, and practices (KAP) of beef cattle farmers regarding LSD. A total of 384 farmers provided data on their sociodemographic characteristics and responses to KAP-related questions. Path analysis was employed to examine how KAP components interact and how demographic factors influence these relationships. The analysis revealed strong positive relationships between farmers’ knowledge and attitudes (β = 0.96, p < 0.001) and between attitudes and practices (β = 0.08, p < 0.008). Farmers with greater knowledge of LSD were more likely to adopt positive attitudes toward disease control and implement effective management practices. Additionally, knowledge had a direct influence on farmers’ practices (β = 0.38, p < 0.001), indicating that improved awareness of LSD is linked to better disease management. By strengthening knowledge, these efforts can positively influence attitudes and practices, leading to more effective control strategies. As the first KAP study on LSD in Thailand, this research highlights the critical role of targeted education programs in improving farmers’ understanding of LSD. Ultimately, enhancing disease management through education can help reduce the economic impact of LSD on Thailand’s livestock sector and promote sustainable farming practices.

Abstract Sheep and goat coccidiosis has a worldwide distribution and is an important disease on lambing farms. Infection with multiple Eimeria species can lead to severe intestinal damage in sheep/goats and economic losses on farms. Disease is a serious constraint to the healthy development of small ruminant farming. Studies published on PubMed, CNKI, VIP, Wanfang and the resulting references of selected studies were included. Risk factors affecting prevalence were analyzed and stratified by geographic location and climatic variables, age, sex, feeding model, season, sample year, breeds and environment. The total prevalence of coccidia in sheep and goats reached more than 60% in most regions, in which the dominant species in sheep were mainly E. parva, E. ovinoidalis, and E. parva and E. ahsata. East China had the lowest prevalence of coccidia infection in sheep (43.24%), and the dominant species were different from those in the other regions, mainly E. bakuensis and E. gonzalezi. Southwestern China and Central China had slightly less than 40.0% goat coccidia infection, and the dominant species in goats were mainly E. arloingi, E. aligevi, E. hirci and E. ninakohlyakimovae. Sheep/goats of different ages can be infected with coccidiosis, but lambs between 1 and 3 months of age are more susceptible to the disease. When lambs become infected, the pathogen spreads rapidly throughout the herd. Spring, summer and autumn are the seasons with a high incidence of this disease. Environmental pollution may be a significant factor in the development of coccidiosis in sheep raised in large-scale housing. This study provides a comprehensive overview of the species, morphology and geographic distribution of Eimeria species in sheep and goats, summary prevalence in different regions of China, risk factors affecting prevalence, and prevention and control strategies.