Background: Cryptosporidium is a globally distributed zoonotic protozoan parasite in humans and animals. Infection is widespread in dairy cattle, especially in calves, resulting in neonatal enteritis, production losses and high mortality. However, the occurrence of Cryptosporidium spp. in pre- and post-weaned calves in Yunnan Province remains unclear. Methods: We collected 498 fecal samples from Holstein calves on 10 different farms in four regions of Yunnan Province. Nested PCR and DNA sequencing were used to determine the infection, species and genotypes of Cryptosporidium spp. in these animals. Results: The overall occurrence of Cryptosporidium spp. in Holstein calves was 32.9% (164/498), and the prevalence in pre- and post-weaned calves was 33.5% (106/316) and 31.9% (58/182), respectively. Four Cryptosporidium species were identified in these animals, namely C. bovis (n = 119), C. parvum (n = 23), C. ryanae (n = 20) and C. andersoni (n = 2). Based on sequencing analysis of the 60 kDa glycoprotein gene of C. bovis, C. parvum and C. ryanae, six subtypes of C. bovis (XXVIe, XXVIb, XXVIf, XXVIa XXVIc and XXVId), two subtypes of C. parvum (IIdA19G1 and IIdA18G1) and four subtypes of C. ryanae (XXIf, XXId, XXIe and XXIg) were identified. Conclusions: These results provide essential information to understand the infection rate, species diversity and genetic structure of Cryptosporidium spp. populations in Holstein pre-weaned and post-weaned calves in Yunnan Province. Further, the presence of IIdA18G1 and IIdA19G1 in C. parvum implies significant animal and public health concerns, which requires greater attention and more preventive measures.

The zoonotic pathogen Toxoplasma gondii exhibits a diverse global population structure, with a few dominant lineages primarily in the Northern Hemisphere. However, reliance on low-resolution, restriction-based genotyping methods has created a “resolution ceiling,” potentially masking hidden genetic diversity and complex transmission dynamics. In this study, we combined conventional PCR-restriction fragment length polymorphism (RFLP) screening with high-resolution Sanger sequencing targeting 16 genetic markers to unravel the fine-scale epidemiology of 96 T. gondii DNA samples collected from various hosts (including pigs, cats, sheep, birds, bats, and captive wildlife) across 12 provinces and regions in China. Our analysis identifies ToxoDB#9 as the dominant lineage (41/96 samples), revealing substantial intra-clonal diversity within this lineage. We report the North American sylvatic ToxoDB#5 (Haplogroup 12) lineage in a captive caracal in China, documenting the presence of this rare lineage outside its previously recognized range, although its public health significance remains uncertain. Population genetic analyses show high haplotype diversity consistent with clonal diversification with limited geographic structuring. Our study provides an updated baseline for T. gondii genetic diversity in China and supports the value of systematic molecular monitoring within a One Health framework, with whole-genome sequencing required to confirm introduction scenarios, resolve transmission routes, and assess recombination.

Toxoplasma gondii is a food- and environment-borne protozoan that is associated with human infection, food safety and animal health. Despite Yunnan Province being a major food-producing animal region in China, comprehensive data on the seroprevalence and risk factors of T. gondii infection in its food animals remain limited. This study investigated the seroprevalence of T. gondii infection among pigs, sheep and goats, cattle and poultry across all 16 prefectures/cities in Yunnan Province. From April 2023 to December 2024, a total of 10,766 blood samples were collected from clinically healthy livestock and poultry, including 2954 pigs, 1950 cattle, 1961 sheep and goats, and 3901 poultry. Sera were examined for the presence of specific antibodies against T. gondii by the Modified Agglutination Test (MAT). Seroprevalence was calculated, and statistical analyses were conducted to assess risk factors (geographic location, season, and animal species). The overall seroprevalence was 13.7% (1474/10,766), with species-specific rates of 26.3% (516/1961) in sheep and goats, 15.1% (446/2954) in pigs, 12.9% (252/1950) in cattle, and the lowest (6.7%, 260/3901) in poultry. Seroprevalence varied considerably across regions, ranging from 9.4% to 27.5%, with the highest prevalence rate being observed in Diqing (27.5%, 141/515). Based on statistical analysis, season, region and animal species were identified as significant risk factors associated with T. gondii infection in Yunnan Province. Overall, this first province-wide investigation revealed widespread exposure of T. gondii across both regions and species. Stringent and sustained control measures against toxoplasmosis of livestock, poultry, and humans in Yunnan Province are recommended.

Fusarium verticillioides is a common pathogenic fungus of corn since it causes severe yield losses and produces mycotoxins to threaten the health of both humans and livestock. Although extensive research has characterized specific genetic and environmental factors influencing mycotoxin production, a systematic understanding of the temporal transcriptional dynamics governing its developmental progression remains lacking. This study addresses this critical knowledge gap through a time-series transcriptomic analysis of F. verticillioides at four key cultivation stages (3, 5, 7, and 9 days post-inoculation). Transcriptomic analysis identified 1928, 2818, and 1934 differentially expressed genes (DEGs) in the comparisons of FV3 vs. FV5, FV5 vs. FV7, and FV7 vs. FV9, respectively. Gene Ontology enrichment revealed 76, 106, and 56 significantly enriched terms across these comparisons, with “integral component of membrane” consistently being the most enriched cellular component. Pathway analysis demonstrated “amino acid metabolism” and “carbohydrate metabolism” as the most significantly enriched metabolic pathways. Notably, the fumonisin (FUM) and fusaric acid (FA) biosynthetic gene clusters exhibited coordinated peak expression during the early cultivation, followed by progressive decline. Mfuzz clustering further delineated 12 distinct expression trajectories, highlighting the dynamic transcriptional networks underlying fungal adaptation. This work provided the first comprehensive temporal transcriptome of F. verticillioides, establishing a foundational resource for understanding its stage-specific biology and revealing potential time-sensitive targets for future intervention strategies.

Ticks are hematophagous arthropods that parasitize livestock, humans, and wild animals, posing serious threats to public health worldwide. Diverse pathogens have been discovered in ticks, including viruses, bacteria, protozoa, and filarial nematodes; however, ticks also harbor a variety of fungal microbes, which remain largely unexplored. In this study, we isolated and identified culturable fungi associated with ticks collected on cattle and goats in Yunnan Province, China. Of the 1730 ticks obtained, 1690 were identified as Rhipicephalus microplus, and only 40 as Haemaphysalis longicornis via morphology and 16S rRNA gene analysis. A total of 90 fungal strains were isolated from the ticks and identified by morphological examination and ITS sequencing, representing 25 species across 20 genera, with Fusarium being the most prevalent. Among these isolates, Fusarium verticillioides (19 isolates) poses a threat to grains, animals, and humans; Cladosporium cladosporioides (six isolates) and Sarocladium zeae (one isolate) are primarily pathogenic to plants, while Schizophyllum commune (eight isolates), Rhizopus arrhizus (two isolates), Diaporthe phaseolorum (two isolates), and Purpureocillium lilacinum (two isolates) are recognized animal-associated pathogens. The diversity of isolated fungi with documented pathogenicity suggests that ticks may serve as potential vectors for fungal transmission, raising concerns about their role in cross-kingdom disease spread across agriculture and livestock. Furthermore, fungi with entomopathogenic potential, such as P. lilacinum, offer valuable resources for developing biocontrol agents against ticks. Our findings highlight the dual role of ticks as both potential vectors of phytopathogenic fungi and reservoirs of entomopathogenic biocontrol agents. This necessitates a paradigm shift in tick surveillance, integrating mycological monitoring to mitigate agricultural losses and exploring tick-borne fungi for sustainable vector control strategies.