The semen microbiome of miniature pony stallions.

CONTEXT: Little is known about the microbial composition of stallion semen. AIMS: To describe the microbiota detected in equine semen of healthy miniature pony stallions. METHODS: Semen specimens were collected using a Missouri artificial vagina at a single time point. PacBio (Pacific Biosciences) genomic DNA sequencing of the 16S rRNA gene was performed on these specimens, following which next-generation microbiome bioinformatics platform QIIME2 was used to process fastq files and analyse the amplicon data. The data were categorised into genus, family, class, order and phylum. KEY RESULTS: Firmicutes and Bacteroidetes phyla predominated (76%), followed by Proteobacteria (15%). Bacteroidales, Clostridiales and Cardiobacteriales predominated the microbial rank of order (86%). Class was mainly composed of Bacteroidia, Clostridia and Gammaproteobacteria (87%), while family was mainly composed of Porphyromonadaceae , Family_XI and Cardiobacteriaceae (62%). At the level of genus, 80% of the abundance was composed of seven genera, namely Porphyromonas, Suttonella, Peptoniphilus, Fastidiosipila, Ezakiella, Petrimonas and an unknown taxon. CONCLUSIONS: The findings indicate that specific microbiota may be characteristic of healthy miniature pony stallions' semen with some inter-individual variations observed. IMPLICATIONS: Larger equine studies involving fertile and infertile subjects could be informed by this study and could explore the relationship of the semen microbiome to male fertility.

Effect of probiotics and prebiotics on the composition of the equine fecal and seminal microbiomes and sperm quality: A pilot study.

Probiotic and prebiotic effects on equine semen and gastrointestinal microbiome composition and sperm quality are unknown. This study aimed to evaluate the effects of pre-, pro- or synbiotic supplementation on fecal and semen microbiome composition and sperm quality parameters of stallions. This Latin square crossover trial involved four miniature pony stallions receiving control diet only, or addition of a pro-, pre- or synbiotic formulation. Full-length 16S rRNA gene amplicon sequencing was used to measure diversity of semen and fecal microbiomes. Total sperm count, total motility, progressive motility, DNA integrity, lipid peroxidation and mitochondrial oxidative stress, biomarkers of sperm quality, were measured after each intervention. A general linear model was employed to analyse and compare microbiome diversity measures and sperm quality data across four time points. Shannon's diversity index (alpha-diversity), and evenness of semen and gastrointestinal microbiomes were significantly different (p<0.001). A trend was observed for prebiotic effects on the diversity indices of the GI microbiome (p= 0.07). No effects of treatments were observed on either semen microbiome or sperm quality. Pre-, pro- and synbiotic supplements showed no negative effect on sperm quality parameters observed. This proof of concept provides preliminary data to inform future studies exploring the relationship between microbiomes and fertility.

The Safety, Tolerability and Efficacy of Probiotic Bacteria for Equine Use.

Probiotic bacteria are used widely as nutritional supplements and treatment interventions in the management of livestock and companion animals. The aim of this review is to summarize the current evidence reporting on the safety, tolerability and efficacy of probiotic bacteria use in horses. An online search of five databases for studies reporting on the use of probiotic bacteria use in horses which were either healthy or had a gastrointestinal or extraintestinal disease was conducted. A total of 18 articles were eligible for full review. No clear benefits were identified to support supplementation of equids with probiotic bacteria to improve starch and fiber digestion, nor for the treatment of colic or prevention of salmonellosis. Conflicting results were seen with the management of scouring in neonatal foals. Exacerbation of diarrhea and additional adverse events were reported in response to the administration of high doses of novel probiotic bacterial species. Probiotic bacteria given to exercising horses, improved aerobic fitness and stamina. The majority of probiotic bacterial species used in equine studies are bacterial species commonly used for human consumption and indigenous to the human gastrointestinal microbiota. There is a paucity of evidence to support the use of probiotic bacteria in the health maintenance and disease management of horses. While there are unclear and conflicting results associated with probiotic bacteria use for gastrointestinal conditions in both horses and foals, the administration of multistrain bacterial formulations to increase stamina in exercising horses shows promise.