Tibetan sheep have unique intestinal microorganisms in their intestines that are adapted to the highland alpine and anoxic environment. To further clarify the probiotic properties of Tibetan sheep-derived probiotics, we selected three Tibetan sheep-derived probiotic isolates (Enterococcus faecalis EF1-mh, Bacillus subtilis BS1-ql, and Lactobacillus sakei LS-ql) to investigate the protective mechanisms of monocultures and their complex strains against Clostridium perfringens type C infection in mice. We established a model of C. perfringens type C infection and used histology and molecular biology to analyze the effects and mechanisms of different probiotic treatments on mice after C. perfringens type C infection. After supplementation with either probiotics or complex probiotics, mice were improved in terms of weight reduction and reduced the levels of cytokines in serum and increased the levels of intestinal sIgA, and supplementation with complex probiotics was effective. In addition, both probiotic and complex probiotic supplementation effectively improved the damage of intestinal mucosa and spleen tissue. The relative expressions of Muc 2, Claudin-1, and Occludin genes were increased in the ileum. The three probiotics and the compound probiotics treatment significantly reduced the relative mRNA expression of toll-like/MyD88/NF-κB/MAPK. The effect of probiotic treatment was similar to the results of engramycin treatment, but the effect of engramycin treatment on intestinal sIgA was not significant. Our results clarify the immunomodulatory effects of the three probiotic isolates and the complex probiotics on C. perfringens infection, and the repair of the intestinal mucosal barrier.

In this study, we screened differentially expressed genes in a multidrug-resistant isolate strain of Clostridium perfringens by RNA sequencing. We also separated and identified differentially expressed proteins (DEPs) in the isolate strain by two-dimensional electrophoresis (2-DE) and mass spectrometry (MS). The RNA sequencing results showed that, compared with the control strain, 1128 genes were differentially expressed in the isolate strain, and these included 227 up-regulated genes and 901 down-regulated genes. Bioinformatics analysis identified the following genes and gene categories that are potentially involved in multidrug resistance (MDR) in the isolate strain: drug transport, drug response, hydrolase activity, transmembrane transporter, transferase activity, amidase transmembrane transporter, efflux transmembrane transporter, bacterial chemotaxis, ABC transporter, and others. The results of the 2-DE showed that 70 proteins were differentially expressed in the isolate strain, 45 of which were up-regulated and 25 down-regulated. Twenty-seven DEPs were identified by MS and these included the following protein categories: ribosome, antimicrobial peptide resistance, and ABC transporter, all of which may be involved in MDR in the isolate strain of C. perfringens. The results provide reference data for further investigations on the drug resistant molecular mechanisms of C. perfringens.

Hippophae rhamnoides subsp. sinensis Rousi has high ecological and medicinal value, and it is an important plant resource unique to the arid regions of Northwest China. Exploring the influence of climate characteristics and soil factors on the composition, diversity, and function of the rhizosphere bacterial community of Chinese seabuckthorn is of great value for developing and popularizing characteristic plant resources in the arid regions of Northwest China. In this study, the rhizosphere soil of 13 Chinese seabuckthorn distribution areas in the northwest of China was taken as the research object, the bacterial community map was constructed based on 16S rRNA gene high-throughput sequencing technology, and the species abundance composition, structural diversity, molecular co-occurrence network, and phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt), as well as the function of rhizosphere soil bacterial community, were systematically studied. Combined with Mantel test and redundancy analysis (RDA), the key habitat factors driving the rhizosphere soil bacterial community structure of Chinese seabuckthorn were explored. The results showed that: (1) The number of amplicon sequence variants (ASVs) in rhizosphere soil bacterial community of Chinese seabuckthorn was the highest in S2(3072) and the S12(3637), and the lowest in the S11(1358) and S13(1996). The rhizosphere soil bacterial community was primarily composed of Proteobacteria, Actinobacteriota, and Acidobacteriota. Except for the S6 and S11 habitats, the dominant bacterial genera were mainly Achromobacter, Acidobacter (RB41), and Sphingomonas. (2) The α and β diversity of rhizosphere soil bacterial communities of Chinese seabuckthorn across 13 distribution areas were significantly different. The number of operational taxonomic units (OTUs), Ace index, and Chao 1 index of soil bacterial community in the S12 distribution area are the highest, and they are the lowest in S11 distribution area, with significant differences. The aggregation of soil bacterial communities in the S5 and S10 distribution areas is the highest, while it is the lowest in the S6 and S11 distribution areas. (3) PICRUSt function classification of soil bacteria showed that Metabolism and Genetic Information Processing functions were the strongest across all distribution areas, with S10 exhibiting higher functional capacity than other areas and S11 showing the weakest. (4) Cluster analysis revealed that soil bacteria across the 13 distribution areas were clustered into two groups, with S10 and S12 distribution areas as one group (Group 1) and the remaining 11 distribution areas as another group (Group 2). (5) Redundancy analysis revealed that pH was the key soil environmental factor driving the rhizosphere soil bacterial community α-diversity of Chinese seabuckthorn, followed by altitude (ALT) and soil water content (SWC). In summary, Chinese seabuckthorn prefers neutral to alkaline soils, and environmental factors play an important role in driving bacterial diversity, community structure, functional profiles, and co-occurrence networks in rhizosphere soil of Chinese seabuckthorn.

Tibetan sheep play a vital role in the livelihoods of herders and are an important part of the ecosystem of the Tibetan Plateau. In order to study the characteristics of the gut microorganisms of Tibetan sheep at high altitude, this study employed macrogenomic techniques to analyse the diversity and differences in the gut flora of Tibetan sheep in different regions of high altitude and high cold. The results demonstrated that at the phylum level, the dominant phylum in the ileo-cecum segment of Tibetan sheep in Qilian, Henan and Gonghe counties was identical, namely Euryarchaeota, Firmicutes and Ascomycota. At the level of the archaebacterial genus, the dominant bacteria of the ileocecal segment of Tibetan sheep in Qilian County, Henan County and Gonghe County were Methanobrevibacter. At the level of bacterial genus, the dominant bacteria of Tibetan sheep ileocecal in Qilian County and Henan County were Bacteroides, while in Gonghe County, the dominant bacteria were Bifidobacterium. At the level of fungal genus, there were notable differences in the abundance of Tibetan sheep ileocecal genus across different regions. However, the abundance of cecum genus exhibited a more consistent trend across regions. From the perspective of functional prediction, the metabolic pathways enriched in the intestinal segments of Tibetan sheep in different regions were found to be identical, with the relative abundance of each functional gene also being essentially uniform. This result will provide a foundation for further research on the mechanism of action of gut microbes in ruminants at high altitude and alpine regions.