Brucella transfers effectors into host cells, manipulating cellular processes to its advantage; however, the mechanism by which effectors regulate cellular processes during infection is poorly understood. A growing number of studies have shown that apoptosis and autophagy are critical mechanisms for target cells to cope with pathogens and maintain cellular homeostasis. BtpB is a Brucella type IV secretion system effector with a complex mechanism for manipulating host infection. Here, we show that the ectopic expression of BtpB promoted DNA fragmentation. In contrast, an isogenic mutant strain, ΔbtpB, inhibited apoptosis compared to the wild-type strain B. suis S2 in RAW264.7 cells. In addition, BtpB inhibited autophagy, as determined by LC3-II protein levels, the number of LC3 puncta, and p62 degradation. We also found that BtpB reduced autophagolysosome formation and blocked the complete autophagic flux. Moreover, our results revealed that the autophagy inhibitor, chloroquine, reduces Brucella’s intracellular survival. Overall, our data unveil new mechanisms of virulence implicating the effector BtpB in regulating host intracellular infection.

The gut bacterial community is essential for maintaining lifelong health and productivity in ruminants, but the relationship between the gut microbiota and the sustained productivity of ruminants remains inadequately understood. In this study, we selected long-lived dairy cows in mid-lactation (≥5 parities) with different levels of milk production (n = 10). Significant differences were observed in the rumen bacterial structures between the two groups of dairy cows, whereas no significant differences were detected in the rectum bacterial communities. Additionally, there were no significant differences in serum oxidative stress biomarkers, inflammatory markers, or immunological markers between the long-lived high-yield (LH) and long-lived low-yield (LL) dairy cows. Furthermore, the concentrations of propionate (Pr) in the rumen and butyrate (Bu) in the rectum were elevated in the high-yield group. Spearman correlation and microbial co-occurrence network analyses revealed that several rumen-enriched bacteria, such as Syntrophococcus, Lachnospira, Shuttleworthia, Erysipelotrichaceae_UCG-2, and Roseburiaare associated with rumen propionate (Pr) production. In the rectum, the reduced abundance of Christensenellaceae_R-7_group and Moryella favors butyrate production. Furthermore, Random Forest machine learning analysis demonstrated that six bacterial taxa in the rumen combined with one serum biomarker, as well as three bacterial taxa in the rectum combined with three serum biomarkers, can serve as potential biomarkers for distinguishing between LH and long- LL dairy cows, achieving prediction accuracies of 92 and 99%, respectively. The findings of this study indicate that rumen and rectum bacteria are associated with the milk production phenotypes of dairy cows with sustained productivity. The rumen microbes are closely linked to the long-term productive capacity of dairy cows and represent a key target for the development of gut microbiota-based interventions. The unique bacterial communities of the rumen and rectum of long-lived high-yielding dairy cows contribute to maintaining their productive capacity.

Brucella, a zoonotic facultative intracellular pathogenic bacterium, poses a significant threat both to human health and to the development of the livestock industry. Alanine racemase (Alr), the enzyme responsible for alanine racemization, plays a pivotal role in regulating virulence in this bacterium. Moreover, Brucella mutants with alr gene deletions (Δalr) exhibit potential as vaccine candidates. However, the mechanisms that underlie the detrimental effects of alr knockouts on Brucella pathogenicity remain elusive. Here, initially, we conducted a bioinformatics analysis of Alr, which demonstrated a high degree of conservation of the protein within Brucella spp. Subsequent metabolomics studies unveiled alterations in amino acid pathways following deletion of the alr gene. Furthermore, alr deletion in Brucella suis S2 induced decreased resistance to stress, antibiotics, and other factors. Transmission electron microscopy of simulated macrophage intracellular infection revealed damage to the cell wall in the Δalr strain, whereas propidium iodide staining and alkaline phosphatase and lactate dehydrogenase assays demonstrated alterations in cell membrane permeability. Changes in cell wall properties were revealed by measurements of cell surface hydrophobicity and zeta potential. Finally, the diminished adhesion capacity of the Δalr strain was shown by immunofluorescence and bacterial enumeration assays. In summary, our findings indicate that the alr gene that regulates amino acid metabolism in Brucella influences the properties of the cell wall, which modulates bacterial adherence capability. This study is the first demonstration that Alr impacts virulence by modulating bacterial metabolism, thereby providing novel insights into the pathogenic mechanisms of Brucella spp.

In high-producing dairy systems, the average productive lifespan of cows is around 2.5–4 years. Persistent productivity and longevity are ke

In ruminants, conceptus elongation is a crucial developmental process that depends on uterine lumen fluid (ULF) and coincides with a period of high pregnancy loss. Prostaglandins (PGs) play indispensable roles in conceptus elongation and implantation. However, the effects of uterus-derived PGs on the uterine environment remain unclear. To explore the metabolic pathways and metabolites induced by endometrium-derived PGs that may affect conceptus elongation and implantation in dairy cows, we investigated the biochemical composition of ULF following intrauterine perfusion of meloxicam from days 12 to 14 of the estrous cycle. Intrauterine administration of meloxicam significantly downregulated the prostaglandin-related metabolites in the ULF. A total of 385 distinct metabolites, primarily clustered within lipids and lipid-like molecules, organic acids and derivatives, organoheterocyclic compounds, and benzenoids, were identified. The metabolite network analysis identified 10 core metabolites as follows: S-adenosylhomocysteine, guanosine, inosine, thymidine, cholic acid, xanthine, niacinamide, prostaglandin I2, 5-hydroxyindoleacetic acid, and indoleacetaldehyde. The pathway enrichment analysis revealed three significantly altered metabolic pathways: arachidonic acid metabolism, tryptophan (Trp) metabolism, and linoleic acid metabolism. A total of five metabolites—guanosine, inosine, thymidine, butyryl-l-carnitine, and l-carnitine—were associated with attachment and pregnancy loss and could serve as predictors of fertility. This global metabolic study of ULF enhances our understanding of histotroph alternations induced by uterus-derived PGs during diestrus in dairy cows, with implications for improving dairy cow fertility.