Achieving a balance between enhancing both humoral and cellular immunity while ensuring sustained release for attenuated live vaccines has long been a challenge in the vaccine industry. However, the incorporation of chemical materials as nanoadjuvants has emerged as a promising strategy to address this challenge. In this study, a novel nano-adjuvant with sustained release properties was developed by combining polyethylene glycol (PEG) and other biochemical and molecular biology agents with CpG. The efficacy of this adjuvant was assessed using the widely applied CSF attenuated live vaccine. The body temperature and relative daily weight gain results demonstrated the safety of the formulations. Importantly, the combination of CpG immune enhancers and sustained-release agents led to a synergistic augmentation of both humoral and cellular immune responses. Specifically, the combination of PEG20000 and benzoic acid, along with Seppic white oil, significantly enhanced humoral immunity. In contrast, the combination of total white oil, PEG6000 and benzoic acid demonstrated superior efficacy in promoting cellular immunity. This study offers important insights into enhancing vaccine efficacy through novel adjuvant combinations, which can be applied to the development of more effective vaccines in the future.

Picornaviruses pose a significant threat to both human and animal health, causing many diseases in humans and swine. Porcine sapelovirus (PSV) is a globally reported enteric picornavirus commonly associated with subclinical or mild enteric infections in swine populations. Critically, unlike other neurotropic picornaviruses such as Enterovirus A71, no PSV strain has been reported to breach the blood–brain barrier (BBB). Since 2023, outbreaks of diarrhea with concurrent neurological signs like ataxia and lameness have been observed in weaned piglets across China, in particular, on a farm in Zhejiang province in 2025 with 100% morbidity and 20% mortality among the cases. Routine diagnostics ruled out common swine pathogens, but qPCR was positive for PSV. We successfully isolated three PSV strains (ZJ, FJ, SD) from affected piglets. Genetic analysis revealed that the PSV-ZJ is a novel recombinant between strains YC2011/2012 (China, 2011) and XTND/2019 (Vietnam, 2018). Pathogenicity assessment confirmed that the recombinant PSV-ZJ is highly pathogenic, causing severe diarrhea, growth retardation, and significant viral shedding via the respiratory and digestive tracts. qPCR and histopathology confirmed viral presence in intestinal and brain tissues, indicating that PSV-ZJ can cross the blood–brain barrier. This study presents the first quantitative viral load and histopathological data for a highly pathogenic recombinant PSV strain in China and emphasizes the critical role of recombination in driving viral virulence evolution, necessitating enhanced surveillance and the development of targeted preventive strategies.