In vitro evaluation of probiotic properties of lactic acid bacteria isolated from the vagina of yak (Bos grunniens)

Abstract

Bovine endometritis is an inflammatory disease of the uterus that occurs after parturition and can result in the destruction of uterine microecology, disruption of hormone secretion, and even infertility. Problems such as antibiotic residues, pathogen resistance, and microbiota dysbiosis caused by conventional antibiotic therapy cannot be ignored. According to the microecological balance theory, probiotics have the potential to prevent or cure endometritis in cattle. Probiotics can positively influence host physiology by regulating microecological imbalance, modulating immunity, and antagonizing pathogens. Since some probiotics contribute to host health only in their specific natural niches, lactic acid bacteria (LAB) from the vagina may have better potential to fight against vaginal and uterine infection. The yak (Bos grunniens) is an ancient and primitive livestock animal that is adapted to high altitude and harsh environments (cold, nutritional deficiencies, and hypoxia). However, to our knowledge, there have been no studies on yak vaginal LAB. Therefore, the purpose of this study was to isolate vaginal LAB from yak, evaluate and compare the probiotic potential and safety of the isolates, and help establish the probiotics library that can be used in the prevention and/or treatment of endometritis. Twenty-five vaginal swabs were collected from healthy yak and cultured in deMan, Rogosa, and Sharpe (MRS) broth. Tentative LAB strains were preliminarily determined through calcium dissolving zone and morphological identification, and the strains were then identified using 16S rRNA gene sequencing. The probiotics of the isolates were detected using cell aggregation, hydrophobicity, resistance to acid and bile salt, adhesion, and antibacterial activities. Additionally, antimicrobial susceptibility, hemolytic activity, and detection of potential virulence factors were determined in order to confirm the safety of these strains. Five isolates were identified: Leuconostoc mesenteroides, Lactobacillus plantarum, Enterococcus hirae, Lacticaseibacillus camelliae, and Lactobacillus mucosae. All isolates had certain growth resistance, aggregation ability, effective antimicrobial potency against Escherichia coli, Staphylococcus aureus, and Salmonella typhimurium, were sensitive to most antibiotics, and could effectively adhere to bovine endometrial epithelial cells (BEECs). None of the isolates showed hemolytic activity or harbored virulence factors. Our results indicated that the five isolates have considerable potential as probiotics that can be used to prevent and/or treat bovine endometritis. We speculate that a mixture of YD6, YD9, and YD25 may yield better results, although this would require extensive experiments to verify.