Schmallenberg virus (SBV) is a transboundary animal pathogen that causes reproductive disorders in ruminants, necessitating standardized molecular surveillance. Adhering to the Chinese national standard GB/T 43159—2023, we developed armored RNA quality control materials using MS2 bacteriophage technology targeting the conserved SBV S segment and comprehensively characterized their physicochemical properties. Transmission electron microscopy (TEM) showed icosahedral symmetric virus-like particles (VLPs, approximately 25 nm), with the stock concentration determined to be 3.48 × 10¹⁰ copies/mL via digital PCR (dPCR). The armored RNA control effectively withstood RNase A degradation and remained stable at 37 °C for over 30 days. In bovine serum matrix simulation assays at identical medium concentrations, the serum armored RNA group (Ct 26.57) was detected 4.89 cycles earlier than the degraded serum naked RNA group (Ct 31.46). This confirms the physical protective efficacy of the MS2 capsid. Concurrently, the serum armored RNA group showed a delay of only 2.17 cycles compared to the aqueous armored RNA group (Ct 24.40), exhibiting a typical matrix effect. Overall, this armored RNA enables full-process quality control encompassing extraction, reverse transcription, and amplification, providing a safe and stable technical reference to support molecular surveillance and diagnostic preparedness for cross-border SBV.

Abortion in cattle entails substantial economic loss, and rapid identification of abortigenic pathogens is critical for timely on-farm response and reduction in human exposure risk. In 2024, two Holstein cows from a small farm in Inner Mongolia aborted in close succession without an obvious cause. Vulvar swabs from both cows, one afterbirth sample, and whole blood from one aborted fetus were collected. Shotgun metagenomic sequencing was performed, followed by host-read removal, taxonomic profiling with Kraken2, de novo assembly of Brucella-aligned reads, and whole-genome comparison. Serological tests, Gram-stained smears, and Brucella genus- and species-specific qPCR assays were used as orthogonal verification. Putative resistance and virulence determinants were screened against CARD and VFDB. Brucella reads were detected in all samples, with the highest relative abundance in the 138-afterbirth (96%). qPCR assays detected Brucella DNA and B. abortus-specific signals in all four samples. A draft Brucella genome was assembled from the 138-afterbirth sample and was phylogenetically placed within B. abortus, showing relatedness to previously circulating Chinese lineages. Cows 138 and 198 were RBT-positive with SAT titres of 1:100 (++). No acquired Brucella resistance genes were identified in CARD. Within 72 h of sample receipt, B. abortus was reported to the farm and local authorities and emergency biosecurity measures were implemented. This field investigation shows that metagenomic sequencing, when combined with conventional serology, microscopy, and targeted qPCR, can support rapid etiological investigation when culture is delayed, hazardous, or biosafety level 3 facilities are unavailable.

Emerging infectious diseases pose a significant threat to global public health and economic security, with zoonoses accounting for a substantial proportion. Livestock such as cattle and sheep are critical reservoirs for zoonotic pathogens and play a key role in transmitting these pathogens to humans and other animals, including dogs and wildlife, due to their close interaction with diverse populations. In this study, we introduce Zoonosis (http://zoonosis.cn/zoonosis/), a comprehensive database that integrates data on pathogens, including Brucella, Mycobacterium tuberculosis, and Bacillus anthracis. Currently, Zoonosis integrates over 4500 samples from more than 60 countries, with a total data volume of 1.8TB, providing a global perspective on zoonotic disease distribution. Equipped with user-friendly visualization and analysis tools, Zoonosis enables rapid biological data interpretation, aiding disease diagnosis and prevention. This resource supports virology, zoology, and epidemiology experts in monitoring cross-species transmissions and mitigating future zoonotic outbreaks. Database URL: http://zoonosis.cn/zoonosis/