Background: 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. Methods: 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. Results: Our analysis identifies ToxoDB#9 as the dominant lineage (41/96 samples), revealing substantial intraclonal 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. Conclusion: 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.

Ageratina adenophora is a globally invasive perennial herb that poses a significant threat to biodiversity. However, recent studies highlight its potential for ecological utilization, demonstrating antibacterial, anti-inflammatory, and insecticidal properties. Research on its effect on Ascaris suum eggs and the underlying molecular mechanisms remains limited. This study investigated the impact of A. adenophora aqueous extract on A. suum egg development and explored key responsive pathways and genes via RNA sequencing (RNA-Seq). A. suum eggs were treated in vitro with various concentrations (0.10%, 0.25%, 0.50%, 1.00%, 2.50%, 5.00%) of the extract. Embryonation rates decreased dose-dependently, with the 5.00% treatment showing the strongest inhibition (34.24%). Eggs treated with 2.50% and 5.00% extract caused significantly less severe pulmonary lesions and lower larval counts in infected mice, indicating reduced infectivity. Transcriptome analysis of control (WH), 0.10% (EH), and 0.50% (MH) treated eggs identified 281 and 1,083 differentially expressed genes (DEGs) for EH vs. WH and MH vs. WH, respectively. GO and KEGG enrichment analyses revealed that DEGs were primarily involved in transmembrane transport, catalytic activity, carbohydrate metabolic processes, the drug metabolism-cytochrome P450 pathway, and glycolysis/gluconeogenesis. Key genes, including CYP44A1, CYP4V2, CYP4C3, and GST-4, were significantly downregulated. qRT-PCR validated the RNA-Seq results. These findings suggest that A. adenophora extract inhibits A. suum egg development, potentially by disrupting energy metabolism and xenobiotic detoxification pathways. This study provides a theoretical basis for developing A. adenophora-based agents targeting the egg stage of A. suum.

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.

Migratory birds are potential carriers for the cross-regional spread of zoonotic diseases due to their high mobility and extensive geographical activity range. Blastocystis, a ubiquitous parasitic protist, can infect a broad spectrum of animal hosts including humans, constituting a potential public health hazard. Currently, data on Blastocystis infection in black-headed gulls (Chroicocephalus ridibundus) remain scarce. This study aimed to investigate the fecal sample positivity rate, epidemiological characteristics, and subtype distribution of Blastocystis in wintering black-headed gulls in Kunming, China. A total of 245 fresh fecal samples were collected from black-headed gulls in Kunming, Yunnan Province, during December 2025 and January 2026. Fecal DNA was extracted and subjected to PCR amplification targeting the small subunit ribosomal RNA (SSU rRNA) gene of Blastocystis. Results showed a 3.27% (8/245) overall Blastocystis fecal sample positivity rate in wintering black-headed gulls, with two subtypes detected: dominant zoonotic pathogenic ST7 (87.5%, 7/8) and ruminant-adapted ST26 (12.5%, 1/8). This study represents the first report of Blastocystis in black-headed gulls in China, providing data supporting the understanding of genetic subtype diversity of Blastocystis in avian hosts.

Toxoplasma gondii is a significant foodborne parasite. However, the precise thermal conditions required to inactivate its tissue cysts in meat remain poorly defined. This study systematically determined the effects of temperature (45-70 °C) and time (10-30 min) on cyst viability. Cysts treated under each condition were orally administered to susceptible C57BL/6J mice, and infectivity was comprehensively assessed through survival, clinical signs, serology (IgG), qPCR, and histopathology. Results demonstrated that treatment at 60 °C for 10 min or under more stringent conditions completely abolished infectivity, as evidenced by 100% survival, the absence of specific antibodies, and the non-detection of parasite DNA or lesions in tissues. Thus, 60 °C for 10 min is established as a critical inactivation threshold, providing a definitive reference for developing science-based thermal processing guidelines to enhance meat safety.

BACKGROUND:Eucoleus annulatus is a parasitic nematode that inhabits the upper digestive tract of avian hosts, posing significant threats to avian health and poultry production. However, the gene information and gene expression characteristics underlying its physiological specialization and parasitic adaptation remain poorly understood. METHODS:In this study, we applied an integrated transcriptomic approach to generate a high-quality full-length transcriptome of E. annulatus using PacBio Iso-Seq and to characterize body section-specific gene expression patterns using Illumina RNA sequencing (RNA-Seq). Differentially expressed transcripts (DETs) among its head, middle, and tail sections were identified, and their functional annotations were assessed through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. RESULTS:PacBio Iso-Seq generated 21,951 high-confidence, non-redundant full-length transcript isoforms, among which 6921 were annotated in the Nr, Pfam, COG, KEGG, and GO databases. Comparative RNA-Seq analysis revealed pronounced section-specific transcriptional divergence, with 1570, 1533, and 1600 DETs enriched in the head, middle, and tail sections, respectively. DETs in the head were significantly enriched in pathways related to amino acid metabolism, RNA processing, and ion transport, while the DETs in the middle body section were primarily associated with glycolysis, oxidative phosphorylation, and transcriptional regulation, indicating elevated metabolic and transcriptional activity. DETs in the tail were significantly enriched in processes related to protein degradation, structural maintenance, and stress adaptation, suggesting roles in environmental response and physiological resilience. CONCLUSIONS:This study, for the first time, reports the first full-length transcriptome of E. annulatus and reveals distinct gene expression profiles across different body sections. These findings provide valuable molecular insights into the spatial organization of gene expression in E. annulatus and establish a foundation for studying its biology and host-parasite interactions in the future.

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.

BACKGROUND:While Toxoplasma gondii infection poses a significant health threat, its impact on the localized intestinal lipid metabolism remains unclear. METHODS:Thus, this study established an acute infection model in BALB/cJ mice and analyzed the colonic contents collected 10 days post-infection via untargeted lipidomics. The lipid profiles of the two groups diverged substantially, with a clear separation evident between infection and control conditions. RESULTS:We identified 65 upregulated and 87 downregulated differential lipids, primarily falling into the glycerophospholipids and sphingolipids categories. Pathway enrichment analysis revealed that the choline metabolism in cancer and glycerophospholipid metabolism pathways was pinpointed as being among the most perturbed following infection. Correlation and network analyses further suggested that key molecules, such as LPC (20:4) and LPA (18:0), may mediate these metabolic pathway abnormalities by regulating related enzymatic activities. CONCLUSIONS:This systematic characterization of the intestinal lipid metabolic landscape in mice during acute T. gondii infection revealed the host intestinal lipid metabolic reprogramming induced by T. gondii infection. The findings offer a novel metabolic perspective for understanding T. gondii pathogenesis and host-parasite interactions.

Cryptosporidium spp. are common protist pathogens, and the growing popularity of pet rodents raises concerns about their potential role in zoonotic parasites transmission. However, epidemiological data on Cryptosporidium spp. in pet rodents in Yunnan Province is scarce. To examine the occurrence of Cryptosporidium spp. in pet rodents in Yunnan, we collected 762 fecal samples from four rodent species across four cities. Nested PCR and DNA sequencing were used to characterize the species and subtypes of Cryptosporidium. The occurrence of Cryptosporidium spp. in guinea pigs (Cavia porcellus), Siberian dwarf hamsters (Phodopus sungorus), Syrian hamsters (Mesocricetus auratus) and fancy rats (Rattus norvegicus domestica) was 18.7% (80/426), 17.3% (36/207), 12.5% (15/120), 0% (0/9), respectively, with an overall rate of 17.2% (131/762). According to regions, the positivity rate of Cryptosporidium spp. in Zhaotong city, Kunming city, Yuxi city and Qujing city was 21.0%, 17.9%, 16.8% and 10.5%, respectively. In terms of sampling location, the occurrence of Cryptosporidium spp. in pet markets, farms and shops was 19.5%, 18.6% and 0%, respectively. Sequence analysis of the small subunit ribosomal RNA (SSU rRNA) gene identified six Cryptosporidium species/genotypes: Cryptosporidium homai (n = 52), Cryptosporidium wrairi (n = 30), Cryptosporidium sp. hamster genotype (n = 25), Cryptosporidium andersoni (n = 20), Cryptosporidium parvum (n = 5), and Cryptosporidium muris (n = 1). Further subtyping of C. andersoni isolates using multilocus sequence typing (MLST) revealed a single subtype, with all isolates identified as A3A4A2A2. All five C. parvum isolates were identified as subtype IIdA15G1 based on the gp60 gene. Our findings demonstrated the presence of the zoonotic C. parvum IIdA15G1 subtype in pet rodents, suggesting that these animals, particularly hamsters, may serve as reservoirs for human-pathogenic Cryptosporidium species. These results underscore the need for improved biosecurity and husbandry practices in the pet rodent trade to mitigate public health risks.

Enterocytozoon bieneusi and Giardia duodenalis are major zoonotic pathogens that often cause diarrhea in immunocompromised humans or animals. Beef cattle are important reservoirs for these two pathogens. Yunnan Province is a major region for beef cattle farming, and its suitable climatic conditions facilitate the transmission of the pathogens. However, research on the prevalence and distribution of E. bieneusi and G. duodenalis in beef cattle in Yunnan remains understudied. This study collected 529 fecal samples from seven beef cattle breeds in four regions in Yunnan Province for an epidemiological survey. Nested PCR combined with sequencing was used to detect E. bieneusi and G. duodenalis, and the sequencing results were analyzed to determine genotypes or assemblage types. Our results demonstrate that the prevalence rates of E. bieneusi and G. duodenalis were 3.0% (16/529) and 3.6% (19/529), respectively. The study identified four genotypes of E. bieneusi, including I (n = 8), J (n = 4), BEB8 (n = 3), and BEB4 (n = 1). Both assemblages E (n = 18) and A (n = 1) were identified among G. duodenalis-positive animals. Phylogenetic analysis revealed that the E. bieneusi genotypes detected in this study belong to Group 2. In conclusion, these findings indicate that although the overall prevalence is relatively low compared to other regions, the presence of zoonotic Group 2 genotypes and assemblage A highlights the potential risk of cross-species transmission. Moreover, the results provide foundational data to support the development of region-specific surveillance and control strategies for bovine giardiasis and microsporidiosis, and emphasize the importance of the One Health approach in managing parasitic infections in livestock populations.

Fusarium verticillioides (F. verticillioides) is an important fungal pathogen known to infect a variety of economically critical crops, particularly maize, causing substantial yield reductions and economic losses worldwide. In addition to its direct damage to agricultural productivity, F. verticillioides threatens public health by producing/secreting potent compounds, including well-known fumonisins (FUMs), which pose significant health threats to both livestock and humans due to their toxicity and carcinogenicity. However, current knowledge of the materials secreted/produced by F. verticillioides, such as secreted proteins and additional secondary metabolites, remains limited. In the present study, we conducted an integrated secretome analysis of F. verticillioides at the exponential growth stage by using proteomic and metabolomic technologies. The results of the present study showed that proteomic analysis identified 185 proteins, including 138 fungus-specific proteins. GO enrichment of these 138 fungus-specific proteins yielded 24 significant terms spanning carbohydrate/polysaccharide and aminoglycan metabolic/catabolic processes, extracellular and membrane-anchored components, and hydrolase/peptidase activities. Meanwhile, KEGG analysis identified starch and sucrose metabolism as the sole significantly enriched pathway. Metabolomic analysis of medium supernatant showed that a total of 2352 metabolites were identified, with 110 unique to the medium supernatant of the fungal group, including fumonisins (A1, B2, B3, B4), fatty acids, and other bioactive compounds. KEGG pathway enrichment highlighted key metabolic pathways, including the TCA cycle, unsaturated fatty acid biosynthesis, and arachidonic acid metabolism. These findings provide new insights into the pathogenic mechanisms of F. verticillioides, suggesting candidates for virulence-associated functions and metabolic adaptations that potentially contribute to its pathogenicity.

Toxoplasma gondii is a widely distributed intracellular parasite that disrupts host immune and metabolic homeostasis. Although accumulating evidence highlights the role of gut microbiota in parasitic infections, the effects of acute T. gondii infection on host gut microbial ecology remain poorly understood. In this study, metagenomic sequencing technology was used to systematically analyze the composition and functional alterations of the ileal microbiota in BALB/c mice on day 10 post-infection. Compared to uninfected controls, T. gondii infected mice exhibited a significant reduction in microbial diversity and a pronounced shift in community structure. Notably, there was an expansion of Proteobacteria, particularly the Enterobacteriaceae family, alongside a marked decline in beneficial taxa such as Actinobacteria and Bacillota. Functional annotation using the KEGG and CAZy databases revealed enrichment of metabolic pathways related to glycolysis/gluconeogenesis, O-antigen nucleotide sugar biosynthesis, bacterial secretion systems, and biofilm formation-Escherichia coli in the infected microbiota. These findings provide novel insights into the dysbiosis of gut microbiota and host-microbe interactions during acute T. gondii infection.

Toxoplasma gondii is an obligate intracellular parasite that causes severe illness in infants infected during pregnancy and in immunocompromised individuals. This parasite manipulates host cells through effector proteins that promote its survival and replication. While the phosphatases in the PP2C family have been shown to regulate host immune responses and contribute to the virulence and pathogenicity of various pathogens, the specific biological functions of PPM3H in T. gondii and its role in host–pathogen interactions remain unclear. In this study, we demonstrate that knockout of ppm3h significantly reduces the virulence and pathogenicity of T. gondii. In contrast, that high expression of ppm3h in the less virulent PRU induced by replacing the ppm3h gene elements of RH strain can enhance its pathogenicity, indicating a direct contribution of PPM3H to virulence in expression-independent manner. Furthermore, PPM3H significantly influenced host gene expression, with differentially expressed genes predominantly enriched in immune and inflammatory pathways. Weighted gene co-expression network analysis identified host immune genes, including chemokines such as Cx3cl1 and Ccl22, as co-expressed with ppm3h. Also, ppm3h co-expressed with T. gondii rhoptry genes including rop18, a well-known virulence factor, suggesting a role for PPM3H in coordinating host–pathogen interactions. Our findings establish that PPM3H enhances T. gondii virulence by modulating the host immune and inflammatory responses. PPM3H does not impact parasite gene expression, invasion or replication in vitro, supporting its role as an immune modulator rather than a general fitness factor. This suggests that T. gondii’s pathogenicity arises not only from immune evasion but also from the active induction of host immune and inflammatory responses mediated by PPM3H.

Cryptosporidium spp. are protozoan pathogens that are widespread within mammals. In recent years, extensive molecular epidemiology studies on Cryptosporidium in dairy cattle have been conducted in Yunnan and worldwide. However, the infection status of these pathogens in beef cattle in Yunnan remains unclear. To examined the occurrence of Cryptosporidium spp. in beef cattle in Yunnan Province, China, we collected 735 fecal samples from six breeds of beef cattle in five regions of Yunnan. Nested PCR and DNA sequencing revealed the infection, species, and genotypes of Cryptosporidium spp. in these animals. The occurrence of Cryptosporidium spp. in Simmental cattle, Brahman cattle, Aberdeen Angus cattle, Yunnan Yellow cattle, Dulong cattle, and Hereford cattle was 32.9% (137/416), 3.8% (4/106), 24.4% (20/82), 3.8% (3/79), 3.2% (1/31), and 0% (0/21), respectively, with an overall rate of 22.4% (165/735). Regarding the regions, the occurrence of Cryptosporidium spp. in Boshan City, Kunming City, Lincang City, Dehong City and Xishuangbanna City was 41.8%, 28.6%, 19.4%, 6.7%, and 3.8%, respectively. In terms of age, the infection rates of Cryptosporidium spp. in pre-weaned, post-weaned, juvenile, and adult cattle were 62.1%, 52.6%, 42.7%, and 7.7%, respectively. According to sex, male cattle were more susceptible to Cryptosporidium infection (28.0%) than females (15.7%). Four Cryptosporidium species were identified in beef cattle: C. andersoni (n = 146), C. bovis (n = 11), C. ryanae (n = 7), and C. occultus (n = 1). Multilocus sequence typing analysis at the MS1, MS2, MS3, and MS16 gene loci revealed four subtype families of C. andersoni (A4A4A4A1, A5A4A4A1, A4A4A2A1, A1A4A4A1). Additionally, sequencing analysis of the 60-kDa glycoprotein gene identified three subtype families of C. bovis (XXVIc, XXVId, XXVIe) and one subtype family of C. ryanae (XXIb). These findings document the occurrence of Cryptosporidium spp. in beef cattle in Yunnan Province for the first time, providing reference data on the distribution, infection rate, species diversity, and genetic structure of these pathogens in China. To effectively reduce the prevalence of Cryptosporidium spp. in beef cattle in Yunnan, the implementation of proper sanitation management, rigorous rodent control, and farmer education programs is crucial. These integrated measures are critical for maintaining herd health, reducing economic losses, and ensuring meat safety across the province.

Toxoplasma gondii is an obligate intracellular protozoan that infects humans and other mammals. The C57BL/6J mouse strain is regarded as an ideal model organism for studying T. gondii due to its susceptibility to T. gondii infection and its other advantages over other laboratory animals. However, systematic studies on the response dynamics of the susceptible C57BL/6J mice after oral infection with T. gondii cysts are lacking. To address this research gap, we investigated the spatiotemporal dynamics of infection, colonization, and antibody fluctuations in susceptible C57BL/6J mice orally infected with Type II T. gondii ME49 strain cysts. Mice were orally challenged with T. gondii cysts to examine the infection dynamics. Daily monitoring was conducted for 60 days post-infection (dpi) to assess animals’ clinical signs and survival rates. The parasite burden in various organs was quantified using qPCR targeting the T. gondii B1 gene. The serum antibody responses were evaluated using ELISA. The cyst burden in the mouse brain was assessed via histology and immunofluorescence. T. gondii infection induced clinical symptoms in the mice, including fever and weight loss. T. gondii rapidly invaded the mice’s small intestine, spleen, lungs, liver, and heart via the bloodstream within 1–5 dpi. T. gondii had breached the blood–brain barrier and colonized the brain by 7 dpi. The levels of Toxoplasma-specific IgG antibodies increased and stabilized for two months (until the experiment ended). Systemic parasite dissemination occurred rapidly, infiltrating most tissues and organs, leading to pronounced enteritis and multi-organ damage due to inflammation. The tachyzoites differentiated into bradyzoites when T. gondii infection progressed from the acute to the chronic phase in mice, forming tissue cysts in organs, including the muscles and brain. As a result, the predilection site of T. gondii in mice is the brain, which is where the cysts persisted for the host’s lifetime and continuously induced meningitis. These findings provide valuable insights into the spatiotemporal diffusion, colonization, predilection sites, temporal antibody dynamics, pathogen detection methodologies, and histopathological changes in C57BL/6J mice following oral infection with T. gondii cysts. These insights are important for elucidating T. gondii’s pathogenesis and host–T. gondii interaction.

Toxoplasma gondii is an obligate intracellular parasite of phylum Apicomplexa that poses a huge threat to pregnant hosts, and induces tragic outcomes for pregnant hosts, fetuses and newborns. However, the molecular mechanism underlying the tragic consequences caused by T. gondii remains to be revealed. In the present study, we applied RNA-seq to study the transcriptomic landscape of the whole reproductive organ of pregnant mice post T. gondii infection, aiming to reveal the key altered biological characters of reproductive organs of pregnant mice that could contribute to the tragic outcomes caused by T. gondii infection. The results of the present study showed that the transcriptome of reproductive organs of pregnant mice was significantly altered by T. gondii infection. A total of 2,598 differentially expressed genes (DEGs) were identified, including 1,449 upregulated genes and 1,149 downregulated genes. Enrichment analysis of the DEGs showed that the significantly altered features of reproductive organs of pregnant mice were excessive inflammatory responses, downregulated metabolism processes, and congenital diseases. The chemotaxis of immune cells in the reproductive organs of infected pregnant mice could also be reshaped by 19 differentially expressed chemokines and 6 differentially expressed chemokine receptors that could contribute to the damages of reproductive organ in pregnant mice. Overall, the findings of present study may help to understand the pathogenic mechanism of the acute T. gondii infection in reproductive organs of pregnant mice, and it could also help to improve toxoplasmosis therapeutics for pregnant individuals.

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.

Toxoplasma gondii, a globally distributed zoonotic obligate intracellular parasite, infects a wide array of mammals, including humans, sheep, and birds. As a unique sheep breed in southwestern China, Yunnan semi-fine wool sheep occupies an important position in animal husbandry in Zhaotong due to its strong adaptability, high reproductive rate, and excellent wool quality. Lambs infected with T. gondii are prone to neurological symptoms and growth retardation, while T. gondii infection in ewes can cause abortions, stillbirths, and deformities, thus affecting sheep reproduction and sheep product quality. Meanwhile, mutton and dairy products contaminated with T. gondii can become potential sources of human infection, potentially threatening public health and safety. To understand the T. gondii infection in semi-fine wool sheep in Zhaotong, Yunnan Province, 586 blood samples were collected and subjected to indirect hemagglutination assay (IHA) for T. gondii antibodies, and the infection-related factors were analyzed through cross-sectional analysis. In the meantime, nested PCR was conducted on a total of 217 samples collected from 31 rodents caught in and around the sheep breeding ground to test the T. gondii B1 gene in rodent tissues. A total of 94 sera tested positive for T. gondii antibodies, with a total positive rate of 16.04

Additional file 2: Table S2. The green tea polyphenol-associated targets obtained from the HERB, TCMSP and PharmMapper databases are shown in sheet 1, and predicted herbs targeting MMP1, MMP7, NOS2 and EPHA2 from the HERB database are shown in sheet 2.

为了解云南省昆明市孔雀感染球虫的种类及种属关系,采集昆明市 2 个动物园孔雀粪便样品及肠道内容物样本进行球虫卵囊的收集,进行形态学鉴定,PCR 扩增球虫ITS rRNA 基因,测序和序列分析,并利用MEGA6.0 软件构建遗传系统进化树.结果显示,扩增的ITS rRNA基因序列长度为 492 bp 和 493 bp,共鉴定出 2 种球虫,分别为和缓艾美耳球虫(Eimeria mitis)和火鸡和缓艾美耳球虫(E.meleagrimitis),相似度均为 100％;进化分析结果表明,孔雀和火鸡之间可能存在球虫的交叉感染风险.该研究结果丰富了野生雉类寄生虫病的资料.