Pasteurella multocida (P. multocida) is a significant pathogenic bacterium that causes serious disease and death in the yaks of the Tibetan Plateau, and the existing inactivated vaccines are limited by low protection and reactogenicity. Outer membrane vesicles (OMVs) derived from a yak-origin serogroup B P. multocida isolate were evaluated as a potential vaccine candidate in the present study. The purified OMVs were characterized by transmission electron microscopy and nanoparticle tracking analysis, which demonstrated the presence of typical bilayer vesicles ranging from 20 to 300 nm in diameter. Proteomic profiling revealed 1213 proteins, with many of them being immunologically relevant outer membrane-associated proteins like OmpA, OmpH, Omp16, OmpW, TbpA and PlpP. The functional enrichment analysis showed that these proteins were linked to translation, membrane structure, transport, metabolism, and pathways of adaptation of bacteria. In vitro OMVs were effectively taken up by RAW264.7 macrophages and stimulated robust expression of inflammatory mediators, such as TNF-α, IL-1β, IL-6, iNOS and IL-10, which is indicative of strong innate immunostimulatory capacity. OMV immunization induced significant antigen specific humoral responses in mice and yaks in vivo. In mice, intramuscular immunization was effective in giving full protection against P. multocida challenge but not intranasal immunization. Histopathology also indicated less tissue damage in vaccinated animals, especially in the lung and liver. These findings, taken together, prove that yak-derived P. multocida OMVs have high immunogenicity and protection capabilities, which show their potential as a next-generation vaccine platform to tackle P. multocida infection.
Background: Microplastics (MPs) and nanoplastics (NPs) are now common in land and water ecosystems. Their spread is an increasing issue from a One Health perspective. These particles end up in soils, water, air, and farm inputs. This poses direct risks to animal health and indirect risks to people who eat animal-derived food. There are also risks from plastic additives and pesticides migrating with these particles in animal-based food. Scope and Approach: This review summarizes how MPs and NPs move in agroecosystems and livestock production. It covers their main sources, such as agricultural plastics, sludge-amended soils, plastic-lined storage, and environmental fallout. It explains how farm animals are exposed, including through feed, water, soil contact, and inhalation. Evidence is condensed for occurrence in manure, tissues, and animal products. The review also highlights key analysis challenges, especially those limiting the assessment of nanoplastic exposure. Key Findings: Field surveys show very different contamination levels in the environment. Agricultural soils range from 0.36 to 42,960 particles/kg. Livestock indicators, like contaminated feed and manure, range from 102 to 105 particles/kg. In free-roaming systems, chicken feces have very high loads, showing trophic transfer in land food chains. A pilot study found plastic particles in pig and cow blood, suggesting some particles cross the gut into the blood. Experimental models link MPs/NPs to oxidative stress, inflammation, mitochondrial dysfunction, metabolic disturbance, and potential reproductive toxicity in livestock and poultry. Conclusions and outlook: Animal-based foods provide a major source of human exposure. MPs and NPs have been observed in milk and poultry products, such as packaged meat and eggs (mean 11.67 ± 3.98 particles/egg). There is still a research gap on raw milk taken directly from the teat and on raw eggs that have not been handled or packaged. This gap makes it hard to identify real contamination sources and control strategies. The review stresses the need for harmonized detection methods (especially for NPs), monitoring from farm to fork, and practical ways to reduce plastic use on farms and minimize contamination during processing, feed handling, and packaging.
Inflammatory bowel disease (IBD) results from dysregulated interactions among gut microbiota, mucosal immunity, and host genetics factors, ultimately leading to chronic inflammation and compromise of the epithelial barrier. Existing therapies remain inadequate for many patients, emphasizing the need for strategies that simultaneously target pathogens, maintain barrier integrity, and modulate host responses. Here, we engineer a living nanomaterial therapeutic by coupling the probiotic Bacillus licheniformis with in situ-grown 7-nm nanosilver to creat a probiotic-nanosilver composite and evaluated its efficacy in TNBS-induced murine colitis. In vivo, Nano-Ag-probiotic enhanced survival in a dose-dependent manner, attenuated weight loss and disease activity, and restored both colon length and histology architecture. Mechanistically, the treatment reinforced epithelial integrity by restoring physiological Wnt/β-catenin signaling, inhibiting GSK3β activation, and re-establishing tight-junction scaffolds including ZO-1, occludin, and claudins. These effects were accompanied by attenuation of pro-inflammatory cytokine signaling and downregulation of CCR2, indicating reduced recruitment of inflammatory monocytes. 16S rRNA sequence analysis revealed that Nano-Ag-probiotic treatment restored microbial diversity, improved Firmicutes/Proteobacteria ratio, reduced Bacteroides and enriched SCFA-associated Dorea in TNBS-induced colitis. Collectively, these data demonstrate that the nanosilver composite rebalances the microbiota-barrier-immunity axis in experimental colitis and represents a translationally promising platform for managing infection-exacerbated intestinal inflammation.
The pathogenesis of inflammatory bowel disease involves interconnected failures in epithelial integrity, immune cell migration, and inflammatory homeostasis in the system, making multifunctional therapeutic approaches a promising option. In this study, we developed a Bacillus licheniformis-templated nano-silver (Nano-Ag) composite within a tannic acid-iron (TA-Fe) metal-phenolic network and evaluated its therapeutic effects in TNBS-induced colitis. The composite exhibited favorable physicochemical properties, including a stable surface architecture and uniform nanosilver deposition, supporting its potential application as a biohybrid nanotherapeutic platform. In vivo treatment produced a dose-responsive protective effect, with the 1 mg formulation showing the most consistent response. This treatment regimen improved survival, reduced morbidity, alleviated diarrhea and fecal bleeding, lowered disease activity, preserved colon length, and significantly restored colonic histoarchitecture. Further histological examination revealed reduced inflammatory infiltration and improved hepatic morphology, indicating protective effects beyond the intestine. At the molecular level, therapeutic efficacy was associated with LEF-1 suppression, axin restoration, CXCR2 and CCR7 attenuation, and CCR5 recovery, consistent with coordinated normalization of Wnt-associated epithelial signaling and chemokine-guided immune trafficking. Serum profiling further revealed broad modulation of dysregulated chemokine networks and partial correction of biochemical disturbances associated with gut-liver and gut-kidney axis dysfunction. Collectively, these findings indicate that the engineered biohybrid nano-Ag platform does not act through a single anti-inflammatory mechanism, but rather through integrated remodeling of epithelial repair, inflammatory signaling, and systemic metabolic stress. Our study identifies TA-Fe-stabilized probiotic nano-Ag therapy as a promising multifunctional strategy for experimental colitis and provides a mechanistic framework for developing next-generation nanomedicines for inflammatory bowel disease.
Ulcerative colitis (UC) is a chronic and progressive inflammatory gastrointestinal disease closely associated with gut microbiota dysbiosis and metabolic homeostasis disruption. Although targeted microbial therapies are an emerging intervention strategy for inflammatory bowel disease (IBD), the mechanisms by which specific probiotics, such as Lactobacillus fermentum 016 (LF), alleviate UC remain unclear. The current study evaluated the effects of LF supplementation on gut health in a basal model using C57BL/6 mice. Subsequently, the preventive effects and mechanisms of LF supplementation on DSS-induced UC were systematically investigated. According to our findings, LF supplementation revealed immunoregulatory capabilities with significantly altered gut the composition of microbiota and metabolic activities, particularly enhancing tryptophan metabolism. In the UC model, LF supplementation effectively mitigated weight loss, increased the disease activity index (DAI), and alleviated diarrhea, rectal bleeding, and colon shortening. Moreover, it reduced colonic pathological damage and histological injury scores. LF intervention improved antioxidant markers and intestinal mucosal barrier function with the activation of the Nrf2–Keap1 signaling pathway and regulation of systemic inflammatory markers, i.e., IL-1β, IL-6, TNF-α, IFN-γ, IL-4, and IL-10. Importantly, LF supplementation reversed metabolic disturbances by significantly increasing the abundance of beneficial genera (e.g., g_Dubosiella, g_Faecalibaculum, g_Odoribacter, g_Candidatus_saccharimonas, g_Roseburia, and g_Eubacterium_xylanophilum_group) and elevating tryptophan metabolites (e.g., melatonin, kynurenic acid, 3-indoleacetic acid, 5-methoxytryptophan, and 5-hydroxyindoleacetic acid). In conclusion, Lactobacillus fermentum 016 exhibits potential for regulating gut microbiota homeostasis, enhancing tryptophan metabolism, and alleviating UC, providing critical insights for developing probiotic-based precision therapeutic strategies for IBD.
Hemorrhagic septicemia (HS) is a fulminant bovine disease across Asia and Africa, yet Pasteurella multocida (P. multocida) isolated from yak is poorly reported. We isolated strain NQ01 from a fatal HS case in Xizang, China and identified it as P. multocida B:2 by morphology, Gram stain, and PCR (kmt1+, bcbD+, LPS L2). NQO1 formed smooth, non-hemolytic colonies. After Gram staining, the cells appeared as red rods with bipolar staining. Antimicrobial testing showed broad susceptibility to β-lactams, aminoglycosides, tetracyclines, fluoroquinolones, midecamycin, florfenicol, polymyxin, and vancomycin, with resistance to metronidazole, trimethoprim sulfamethoxazole, and clindamycin. Streptomycin and ofloxacin had intermediate activity. In mice, the intraperitoneal and intranasal LD50 values were 40.64 CFU/mL and 9.53 × 106 CFU/mL, respectively. The intranasal fatal cases were characterized by bacteremia with multifocal disseminated intravascular coagulation involving lung, liver, and spleen. The complete genome comprises a single 2.33 Mb chromosome (40.47% GC, 2115 CDS, no plasmids) with only one resistance gene (Eco_EFTu_PLV) and 28 virulence genes spanning adhesion (tadA, rcpA, ppdD, pilB, tuf/tufA, htpB, PM_RS00430, PM_RS00425, PM_RS08640), immune modulation (lpxB/C/D, msbB, manB, rfaE/F, gmhA/lpcA, kdsA, pgi, wecA, galE, bexD’, ABZJ_RS06285, ABD1_RS00310), and nutritional/metabolic factor (hgbA, hemR, hemN), plus a YadA-like factor. Phylogenetically, NQ01 clusters with regional B:2 bovine/yak isolates. Collectively, these data define NQ01 as a highly virulent, low-resistance yak isolate and a practical model for natural-route HS pathogenesis and targeted control in high-altitude pastoral settings yaks.
Pasteurella multocida serotype B:2 is a primary agent of hemorrhagic septicemia (HS) in livestock, and the strain NQ01 isolated from yaks highlights its cross-species impact. In this study, a murine intranasal infection model was established using P. multocida NQ01 to assess how acute respiratory infection perturbs gut homeostasis. Mice were intranasally inoculated with NQ01, and at 36 h post-infection, ileal tissues and cecal contents were collected for histopathological examination, 16S rRNA gene sequencing, and untargeted metabolomic analysis. Histopathology revealed obvious acute bronchopneumonia but no overt ileal damage. However, 16S rRNA sequencing of cecal microbiota showed significant dysbiosis: microbial diversity was reduced and community composition shifted, including decreased short-chain fatty-acid-producing taxa and increased opportunistic genera. Metabolomic profiling detected 1444 significantly altered cecal metabolites, and pathway analysis indicated marked disruption of amino acid metabolism, notably the tyrosine metabolism pathway. Key tyrosine pathway metabolites were dysregulated (e.g., elevated L-tyrosine and dopamine with reduced L-DOPA), indicating a breakdown of this metabolic pathway. These findings demonstrate that acute respiratory P. multocida infection profoundly disturbs gut microbiota and metabolism, underscoring disruption of the gut-lung axis. This study provides new insight into the systemic consequences of HS-associated P. multocida infection and offers a basis for exploring the gut-lung interaction in hemorrhagic septicemia pathogenesis.
Gut microbiota and their metabolic processes depend on the intricate interplay of gut microbiota and their metabolic processes. Bacillus licheniformis, a beneficial food supplement, has shown promising effects on stabilizing gut microbiota and metabolites. However, the precise mechanisms underlying these effects remain elusive. In this study, we investigated the impact of polysaccharide-producing B. licheniformis as a dietary supplement on the gut microbiome and metabolites through a combination of scanning electron microscopy (SEM), histological analysis, high-throughput sequencing (HTS), and metabolomics. Our findings revealed that the B. licheniformistreated group exhibited significantly increased jejunal goblet cells. Moreover, gut microbial diversity was lower in the treatment group as compared to the control, accompanied by noteworthy shifts in the abundance of specific bacterial taxa. Enrichment of Firmicutes, Lachnospiraceae, and Clostridiales_bacterium contrasted with reduced levels of Campylobacterota, Proteobacteria, Parasutterella, and Helicobacter. Notably, the treatment group showed significant weight gain after 33 days, emphasizing the polysaccharide's impact on host metabolism. Delving into gut metabolomics, we discovered significant alterations in metabolites. Nine metabolites, including olprinone, pyruvic acid, and 2-methyl-3-oxopropanoate, were upregulated, while eleven, including defoslimod and voclosporin were down-regulated, shedding light on phenylpropanoid biosynthesis, tricarboxylic acid cycle (TCA cycle), and the glucagon signaling pathway. This comprehensive multi-omics analysis offers compelling insights into the potential of B. licheniformis as a dietary polysaccharide supplement for gut health and host metabolism, promising significant implications for gut-related issues.
Pasturella multocida (P. multocida), a gram-negative bacterium, has long been a focus of interest in animal health because of its capacity to cause different infections, including hemorrhagic septicemia. Yaks, primarily found in high-altitude environments, are among the several livestock animals affected by these bacteria. Yaks are essential to the socioeconomic life of the people who depend on them since they are adapted to the cold and hypoxic conditions of highland environments. Nevertheless, these terrains exhibit a greater incidence of P. multocida despite the severe environmental complications. This predominance has been linked to the possible attenuation of the yak’s immunological responses in such circumstances and the evolution of some bacterial strains to favor survival in the respiratory passages of the animals. Moreover, these particular strains threaten other cattle populations that interact with yaks, which might result in unanticipated outbreaks in areas previously thought to be low risk. Considering these findings, designing and executing preventative and control strategies suited explicitly for these distinct biological environments is imperative. Through such strategies, yaks’ health will be guaranteed, and a larger bovine population will be safeguarded against unanticipated epidemics. The current review provides thorough insights that were previously dispersed among several investigations. Its distinct method of connecting the ecology of yaks with the dynamics of infection offers substantial background information for further studies and livestock management plans.
Lumpy skin disease virus (LSDV) is capable of causing transboundary diseases characterized by fever, nodules on the skin, mucous membranes, and inner organs. The disease may cause emaciation with the enlargement of lymph nodes and sometimes death. It has had endemic importance in various parts of Asia in recent years, causing substantial economic losses to the cattle industry. The current study reported a suspected LSDV infection (based on signs and symptoms) from a mixed farm of yak and cattle in Sichuan Province, China. The clinical samples were found positive for LSDV using qPCR and ELISA, while LSDV DNA was detected in Culex tritaeniorhynchus Giles. The complete genome sequence of China/LSDV/SiC/2021 was determined by Next-generation sequencing. It was found that China/LSDV/SiC/2021 is highly homologous to the novel vaccine-related recombinant LSDV currently emerging in China and countries surrounding China. Phylogenetic tree analysis revealed that the novel vaccine-associated recombinant LSDV formed a unique dendrograms topology between field and vaccine-associated strains. China/LSDV/SiC/2021 was found to be a novel recombinant strain, with at least 18 recombination events via field viruses identified in the genome sequence. These results suggest that recombinant LSDV can cause high mortality in yaks, and its transmission might be due to the Culex tritaeniorhynchus Giles, which acts as a mechanical vector.
为建立检测牛传染性鼻气管炎病毒(IBRV)的荧光定量检测方法,本研究根据 IBRV gE基因的序列设计了相应的探针和引物,分别建立TaqMan-MGB探针和SYBR GreenⅠ荧光定量PCR方法,并对2 种方法进行了综合比较.结果显示,TaqMan-MGB探针和SYBR GreenⅠ定量荧光PCR方法的灵敏度分别达到1.0×101 copies/μL和1.0×102 copies/μL;2 种荧光定量方法对除IBRV的其他牛病毒和细菌检测结果均为阴性;重复性检测中变异系数均小于2.3%;检测了130 份来自西藏的牦牛样品,2 种荧光定量PCR阳性检出率均为 100%.以上结果表明,2 种荧光定量方法都可用于检测IBRV,且TaqMan-MGB荧光定量PCR敏感性高于SYBR Green Ⅰ荧光定量PCR.
BACKGROUND:Apoptosis is thought to be involved in all processes, including normal cell cycle, immune system, atrophy, embryonic development, and chemical-induced cellular damage. However, if the normal apoptotic process fails, the results might be disastrous, e.g., chondrocytes damage in tibial dyschondroplasia (TD). TD is a worldwide issue in the poultry sector due to thiram toxicity. Thiram (Tetramethyl thiuram disulfide) is a dithiocarbamate pesticide and fungicide commonly used in horticulture to treat grains meant for seed protection and preservation. PURPOSE:According to prior studies, chlorogenic acid (CGA) is becoming essential for regulating apoptosis. But still, the specific role of CGA in chondrocyte cells remains unclear. The present study explored the molecular mechanism of CGA on chondrocytes' apoptosis with B-cell lymphoma 2 signaling under the effect of miR-460a. METHODS:An in vivo and in vitro study was performed according to our previously developed methodology. Flow cytometry, western blotting, reverse transcription-quantitative polymerase chain reaction, and immunofluorescence assay were used to investigate the involvement of apoptosis and inflammasome related pathways. RESULTS:The CGA decreased the apoptosis rate with the deactivation of miR-460a, accompanied by the activation of Bcl-2. The high expression of miR-460a reduced the cell viability of chondrocytes in vitro and in vivo, that led to the interleukin-1β production. While the apoptotic executioners (caspase-3 and caspase-7) acted upstream in miR-460a overexpressing cells, and its depletion downgraded these executioners. The CGA administrated cells negatively regulated miR-460a expression and thus indicating the deactivation of the apoptotic and inflammasome related pathways. CONCLUSION:Chlorogenic acid had a negative effect on miR-460a, setting off specific feedback to regulate apoptotic and inflammasome pathways, which might be a key feature for chondrocytes' survival.
Antibiotics are often used to promote animal growth and prevent diarrhea in weanling piglets. Nevertheless, intestinal pathogenic bacterial resistance and drug residues caused by antibiotic overuse are worthy of concern and demand an urgent solution. Bacillus amyloliquefaciens TL106 has been isolated from cold- and disease-resistant Tibetan pigs in Linzhi, Tibet.
The present study determined the complete mitochondrial DNA (mt DNA) sequence of Fasciola intermediate (isolated from yaks) based on gene content and genome organization. According to our findings, the genome of Fasciola intermediate was 13,960 bp in length, containing 2 ribosomal RNA (rRNA) genes, 12 protein-coding genes (PCGs), and 22 transfer RNA (tRNA) genes. The A+T content of genomes was 63.19%, with A (15.17%), C (9.31%), G (27.51%), and T as the nucleotide composition (48.02%). Meanwhile, the results showed negative AT-skew (-0.52) and positive GC-skew (0.494). The AT bias significantly affected both the codon usage pattern and amino acid composition of proteins. There were 2715 codons in all 12 protein-coding genes, excluding termination codons. Leu (16.72%) was the most often used amino acid, followed by Val (12.74%), Phe (10.90%), Ser (10.09%), and Gly (8.39%). A phylogenetic tree was built using Maximum-Likelihood (ML) through MEGA 11.0 software. The entire mt DNA sequence of Fasciola intermediate gave more genetic markers for investigating Trematoda population genetics, systematics, and phylogeography. Hence, for the first time, our study confirmed that yaks on the Qinghai-Tibet plateau have the infestation of Fasciola intermediate parasite.
Lactobacillus fermentum (L. fermentum) YLF016 is a well-characterized probiotic with several favorable characteristics. This study aimed to analyze the probiotic characteristics of L. fermentum and uncover the genes implicated in its potential probiotic ability on the base of its genomics features. The complete genome of L. fermentum YLF016 was found to have a circular chromosome of 2,094,354 bp, and 51.46% G + C content without any plasmid. Its chromosome contained 2,130 predicted protein-encoding genes, 58 tRNA, and 15 rRNAencoding genes. Also, it was found to have many other probiotic properties, such as a high survival rate in the gastrointestinal tract with strong adherence to intestinal cells, antibacterial activity against pathogens, and antioxidant activity. Moreover, the genome sequence analysis demonstrated specific genes coding for carbon metabolism pathway, genetic adaption, stress resistance, and adhesive ability. Further analysis revealed its non hemolytic activity and its non-functional ability of virulence factors. In conclusion, L. fermentum YLF016 possesses many valuable probiotic properties that refer to its potential probiotic ability.
Tibial dyschondroplasia debilities apoptotic and inflammasomal conditions that can further destroy chondrocytes. Inflammasomes are specialized protein complexes that process pro-inflammatory cytokines, e.g., interleukin-1β (IL-1β) and IL-18. Moreover, there is mounting evidence that many of the signaling molecules that govern programmed cell death also affect inflammasome activation in a cell-intrinsic way. During the last decade, apoptotic functions have been described for signaling molecules involving inflammatory responses and cell death pathways. Considering these exceptional developments in the knowledge of processes, this review gives a glimpse of the significance of these two pathways and their connected proteins in tibial dyschondroplasia. The current review deeply elaborates on the elevated level of signaling mediators of mitochondrial-mediated apoptosis and the inflammasome. Although investigating these pathways’ mechanisms has made significant progress, this review identifies areas where more study is especially required. It might lead to developing innovative therapeutics for tibial dyschondroplasia and other associated bone disorders, e.g., osteoporosis and osteoarthritis, where apoptosis and inflammasome are the significant pathways.
Yak (Bos grunniens) inhabit an oxygen-deficient environment at the altitude of 3000 m on the Tibetan Plateau, with a distinctive gut micro-ecosystem. This study evaluated the probiotic potential and physiological property of Bacillus licheniformis and Bacillus pumilus isolated from the gut of yaks. Four strains, two Bacillus licheniformis (named D1 and D2) and two Bacillus pumilus (named X1 and X2), were isolated and identified by 16S rRNA sequencing. All strains had potential antibacterial ability against three indicator pathogens: Escherichia coli C83902, Staphylococcus aureus BNCC186335, and Salmonella enteritidis NTNC13349. The antioxidant activity test showed that D2 sample showed the highest antioxidant activity. Furthermore, all four strains had a higher hydrophobicity, auto-aggregation, acid tolerance, bile tolerance, and antibiotic sensitivity, which all contribute to their survival in the gastrointestinal tract and clinical utility. The animal experimentation (40 KM mice, equally divided into five groups of eight mice each) showed that the strain supplementation not only increased daily weight gain and reduced feed conversion ratio, but also increased the length of the jejunum villi and the value of the V/C (Villi/Crypt). In conclusion, this is the first study demonstrated the probiotic potential of Bacillus licheniformis and Bacillus pumilus isolated from yaks, providing a theoretical basis for the clinical application and development of new feed additives.
Cystic echinococcosis (CE) is a livestock disease caused by a parasite known as Echinococcus granulosus. It is one of the primary cause for illness and poverty especially for herders on the Qinghai–Tibet plateau, China. Meanwhile, the Qinghai–Tibet plateau has been a key area for echinococcosis control in China. Here in current study, we determined the seroprevalence of E. granulosus in ruminants on this region. A total of 2,730 serum samples (1,638 samples from yaks and 1,092 samples from sheep) were collected on the plateau during the period of 2017. The samples were assayed for E. granulosus antibodies by commercial enzyme-linked immunosorbent assay kits. Our results exhibited a prevalence percentage of 52.2% in Tibetan yaks and 38.2% in Tibetan sheep. Moreover, there was more chance of being infected with E. granulosus infection in old animals due to more exposure to contaminated sources of infection. However, no significant difference was observed. Furthermore, we observed that the rainfall and presence of several lakes has increased the risk of CE infection in yaks and sheep in the Qinghai, Qinglong, and Baingoin areas. Hence, with this investigation, it was possible to determine the frequency and distribution of CE in yaks and Tibetan sheep on the Qinghai-Tibet plateau, that laying the groundwork for its prevention and management.
2021年1月18—24日,石首麋鹿国家级自然保护区多头麋鹿(Elaphurus davidianus)死亡.采集组织及粪便样品进行实验室诊断.根据临床症状,组织涂片镜检、病理组织切片观察、细菌分离培养和染色镜检、生化鉴定、细菌16S rDNA和多重PCR鉴定,确诊为A型产气荚膜梭菌(Clostridium perfringens)感染,并根据诊断和药敏试验结果,给出相应的防治建议.