The yak (Bos grunniens) exhibits exceptional regional adaptability, enabling it to thrive in the distinctive ecological niches of the Qinghai–Tibet Plateau. Its survival relies on the intricate balance of its intestinal microbiome, essential for adapting to harsh environmental conditions. Despite the documented significance of bacteria and fungi in maintaining intestinal homeostasis and supporting immune functions, there is still a substantial gap in understanding how the composition and functionality of yak gut microbiota vary along altitude–temperature gradients. This study aims to fill this gap by employing 16S rRNA and ITS amplicon sequencing techniques to analyze and compare the intestinal microbiome of yaks residing at different elevations and exposed to varying temperatures. The findings demonstrate subtle variations in the diversity of intestinal bacteria and fungi, accompanied by significant changes in taxonomic composition across various altitudes and temperature gradients. Notably, Firmicutes, Actinobacteriota, and Bacteroidota emerged as the dominant phyla across all groups, with Actinobacteriota exhibiting the highest proportion (35.77%) in the LZF group. Functional prediction analysis revealed significant associations between the LZF group and metabolic pathways related to amino acid metabolism and biosynthesis. This suggests a potential role for actinomycetes in enhancing nutrient absorption and metabolism in yaks. Furthermore, our findings suggest that the microbiota of yaks may enhance energy metabolism and catabolism by modulating the Firmicutes-to-Bacteroidota ratio, potentially mitigating the effects of temperature variations. Variations in gut bacterial and fungal communities among three distinct groups were analyzed using metagenomic techniques. Our findings indicate that microbial genera exhibiting significant increases in yaks at lower altitudes are largely beneficial. To sum up, our research investigated the changes in gut bacterial and fungal populations of yaks residing across diverse altitude and temperature ranges. Moreover, these results enhance comprehension of gut microbial makeup and variability, offering perspectives on the environmental resilience of dry lot feeding yaks from a microbial angle.

Despite the crucial role of the gut microbiota in different physiological processes occurring in the animal body, reports regarding the gut microbiota of animals residing in different environmental conditions like high altitude and different climate settings are limited. The Qinghai–Tibetan Plateau is renowned for its extreme climatic conditions that provide an ideal environment for exploring the effects of high altitude and temperature on the microbiota of animals. Yaks have unique oxygen delivery systems and genes related to hypoxic response. Damxung, Nyêmo, and Linzhou counties in Tibet have variable altitudes and temperatures that offer distinct settings for studying yak adaptation to elevated terrains. The results of our study suggest that amplicon sequencing of V3-V4 and internal transcribed spacer 2 (ITS2) regions yielded 13,683 bacterial and 1912 fungal amplicon sequence variants (ASVs). Alpha and beta diversity indicated distinct microbial structures. Dominant bacterial phyla were Firmicutes, Bacteroidota, and Actinobacteriota. Genera UCG-005, Christensenellaceae_R-7_group, and Rikenellaceae_RC9_gut_group were dominant in confined yaks living in Damxung county (DXS) and yaks living in Linzhou county (LZS), whereas UCG-005 prevailed in confined yaks living in Nyêmo county (NMS). The linear discriminant analysis effect size (LEfSe) analysis highlighted genus-level differences. Meta-stat analysis revealed significant shifts in bacterial and fungal community composition in yaks at different high altitudes and temperatures. Bacterial taxonomic analysis revealed that two phyla and 32 genera differed significantly (p < 0.05). Fungal taxonomic analysis revealed that three phyla and four genera differed significantly (p < 0.05). Functional predictions indicated altered metabolic functions, especially in the digestive system of yaks living in NMS. This study reveals significant shifts in yak gut microbiota in response to varying environmental factors, such as altitude and temperature, shedding light on previously unexplored aspects of yak physiology in extreme environments.

The Jiani yak is a nationally renowned species that is known for its meat which is rich in various minerals, amino acids, and proteins. The rumen microbiota plays a critical role in gastrointestinal health and feed degradation, contributing proteins, lipids, and volatile fatty acids (VFAs) essential for milk and meat production. However, there is limited knowledge about the microbiota of free-ranging Jiani yaks, especially those with 15 ribs. Rumen fluid samples were collected from yaks with 14 (PL) ribs and 15 (DL) ribs from a slaughterhouse in Jiani County, China. The total DNA of rumen fluid microorganisms was extracted for microbiota sequencing. Our results revealed 643,713 and 656,346 raw sequences in DL and PL animals, respectively, with 611,934 and 622,814 filtered sequences in these two yak groups. We identified 13,498 Amplicon Sequence Variants (ASVs), with 2623 shared between DL and PL animals. The ratio of Bacteroidota to Firmicutes differed between PL (3.04) and DL (2.35) animals. Additionally, 6 phyla and 21 genera showed significant differences between yaks with 14 and 15 ribs, leading to altered microbiota functions, with 51 and 35 notably different MetaCyc and KEGG pathways, respectively. Hence, the microbiota of yaks with 15 ribs differs from those with 14 ribs. Therefore, these microbiota-related comparative investigations will provide insights into yak husbandry practices and genetic selection strategies for their improved productivity in harsh environments.

N⁶-methyladenosine (m⁶A), an RNA post-transcriptional modification, plays a crucial role in spermatogenesis. Cattle-yaks are interspecific hybrid offsprings of yak and cattle, and male cattle-yaks are sterile. This study aims to investigate the role of m⁶A modification in male cattle-yak infertility. Herein, testicular tissues were analyzed via histological observations, immunohistochemical assays, reverse-transcription quantitative polymerase chain reaction, Western blotting, and immunofluorescence assays. The results revealed that male cattle-yaks presented smaller testes (5.933 ± 0.4885 cm vs. 7.150 ± 0.3937 cm), with only single cell layers in seminiferous tubules, and weakened signals of m⁶A regulators such as METTL14 (methyltransferase-like 14), ALKBH5 (alpha-ketoglutarate-dependent hydroxylase homolog 5), FTO (fat mass and obesity-associated protein), and YTHDF2 (YTH N⁶-methyladenosine RNA binding protein F2), both at the RNA and protein levels, compared with those of yaks. Altogether, these findings suggest that m⁶A modification may play a crucial role in male cattle-yak sterility, providing a basis for future studies.