Sperm infertility or subfertility is detrimental to the precious highland germplasm like yak whose population has been gradually declining in India. Understanding the ‘omic’ landscape of infertile or subfertile yak sperm can reveal some interesting insights. In an attempt to do the same, this study considered the semen of infertile or subfertile yak bulls for whole-genome and transcriptome evaluations. DNA sequencing revealed that the yak sperm genome contains the necessary genes to carry out all the important biological processes related to the growth, development, survival and multiplication of an organism. Interestingly, RNA Seq results highlighted that genes like VAMP7, MYLK, ARAP2 and MARCH6 showed increased expression, while biological processes related to immune response (GO:0043308, GO:0002447, GO:0002278, GO:0043307, GO:0043312, GO:0002283, GO:0043299 and GO:0002446) were significantly overrepresented. These findings hint at a possible role played by immune system in regulating infertility or subfertility in yaks. Further, in-depth studies can validate these findings and help in improving our biological understanding in this area.

Inaccessibility of veterinary and livestock extension services, and shortages of labour and forage could potentially impact the welfare of yaks (Bos grunniens) in Bhutan. The objective of this study was to assess practices relating to the welfare and management of free-ranging yaks in Bhutan and explore variations between different yak-farming regions. We interviewed herders and observed the behaviour and health status of their animals, using an adaptation of the Welfare Quality® protocol, in three yak-farming regions (east, central and west) of Bhutan between October 2018 and January 2019. In total, for 567 cows and 549 calves, integumentary condition, body cleanliness, ocular and nasal discharge, diarrhoea, signs of damage, and gait were scored. In addition, we assessed 324 cows and 272 calves for avoidance distance and examined 324 cows for subclinical mastitis. The behaviour of the herds was observed in six consecutive 20-min blocks with each block divided into two stages. The first stage (5 min) consisted of counting the number of animals eating, lying down, standing idle and walking. The second stage (15 min) consisted of counting the number of events of agonistic, allogrooming, flehming, self-licking, rubbing/scratching and playing behaviour. Avoidance distance differed between regions for calves, but not for lactating cows. Integumentary lesions, dirty body areas, nasal discharge, ocular discharge, signs of diarrhoea, subclinical mastitis and lameness were virtually absent. A few instances of agonistic behaviour (6% of all counted behavioural events) and flehming behaviour (5% of all counted behavioural events) were observed. Yaks in the central and western regions exhibited more scratching and rubbing behaviour than those in the eastern region. Herders perform a variety of painful management practices (castration, ear tagging, nasal septum piercing) without analgesia, which is a prominent welfare issue. Furthermore, mortality among yaks is relatively high and water sources often dirty, creating a health risk. Nevertheless, the welfare status of yaks living in various regions of Bhutan was assessed as good at the time of visit.

Oxygen concentration influences oocyte quality and subsequent embryo development, but it remains unclear whether oxygen concentrations affect the developmental competence and transcriptomic profile of yak oocytes. In this study, we investigated the effects of different oxygen concentrations (5% versus 20%) on the developmental competence, reactive oxygen species (ROS) levels, glutathione (GSH) content, and transcriptomic profile of yak oocytes. The results showed that a low oxygen concentration significantly increased the maturation rate of yak oocytes (81.2 ± 2.2% vs 75.9 ± 1.3%) and the blastocyst quality of yak in vitro fertilized embryos. Analysis of ROS and GSH showed that a low oxygen concentration reduced ROS levels and increased the content of GSH (75.05 ± 7.1 ng/oocyte vs 50.63 ± 5.6 ng/oocyte). Furthermore, transcriptomic analysis identified 120 differentially expressed genes (DEGs) between the two groups of oocytes. Gene enrichment analysis of the DEGs indicated multiple cellular processes, including oxidative phosphorylation, transcription regulation, mitochondrial regulation, oestrogen signalling pathway, HIF-1 signalling pathway, TNF signalling pathway, were involved in the response to oxygen concentration alterations. Taken together, these results indicated that a low oxygen concentration improved the developmental competence of yak oocytes.

Yaks (Bos grunniens) live primarily in the Qinghai-Tibetan plateau (altitude: 2000–5000 m). Their milk presents unusual characteristics, containing large amounts of solids including fat and protein, and it is, therefore, important to understand the genetic makeup of the yak. To identify potentially critical genes playing a role in yak mammary tissue from colostrum to mature milk phase of lactogenesis, the early lactation (colostrum) stage (ELS; day 1 after parturition) and mature lactation (milk) stage (MLS; day 15) were chosen for comparison. An ELS-specific cDNA library was established by suppression subtractive hybridization and 25 expressed sequence tags at ELS were identified by sequencing and alignment. To further confirm our results the expression levels of 21 genes during the lactation cycle were measured using quantitative real-time RT-PCR (qRT-PCR). The qRT-PCR results confirmed 9 significantly up-regulated genes at ELS vs. MLS in yak mammary tissue, in which the l-amino acid oxidase 1 (LAO1) and collagen, type I, alpha I (COL1A1) were the most significantly up-regulated. During the lactation cycle, the highest expression of some milk fat genes (i.e., XDH and FABP3) in yak mammary tissue appears earlier than that in dairy cow. Our data also indicate MYC potentially playing a central role through putative regulation of COL1A1, CD44, SPARC, FASN and GPAM.

The aim of this research was to identify variation in the yak lipin-1 gene (LPIN1) and determine whether this variation affects milk traits. PCR-single stranded conformational polymorphism (PCR-SSCP) analysis was used to detect variation in the 5′ untranslated region of LPIN1 in 500 yaks from four populations: Tianzhu white yaks, Qinghai yaks, wild × domestic-cross yaks and Gannan yaks. Four unique PCR-SSCP patterns, representing four different DNA sequence variants (named A, B, C and D), were observed. These contained six single nucleotide polymorphisms. Female Gannan yaks with BC genotype produced milk with a higher fat content (P < 0.001) and total milk solids (P < 0.001), than those with the AA, AB and BB genotypes. These results would suggest that LPIN1 is having an effect on yak milk fat synthesis.

Melatonin plays a critical role in several types of cells as an antioxidant to protect intracellular molecules from oxidative stress. The anti-oxidation effect of melatonin in yak embryos is largely unknown. We report that melatonin can protect the development of yak preimplantation embryos against oxidative stress induced by hydrogen peroxide (H2O2). Therefore, the quality of blastocysts developed from zygotes exposed to H2O2 was promoted. In addition, we observed that melatonin reduced H2O2-induced intracellular reactive oxygen species (ROS) levels and prevented mitochondrial dysfunction in zygotes. These phenomena revealed the effective antioxidant activity of melatonin to prevent oxidative stress in yak embryos. To determine the underlying mechanism, we further demonstrated that melatonin protected preimplantation embryos from oxidative damage by preserving antioxidative enzymes. Collectively, these results confirmed the anti-oxidation effect of melatonin in yak embryos that significantly improved the quantity and quality of blastocysts in the in vitro production of embryos in yaks.

This research communication describes the profile of gene expression related to the synthesis of yak milk as determined via quantitative reverse transcription polymerase chain reaction (RT-qPCR). Significant up-regulation during lactation were observed in genes related to fatty acid (FA) uptake from blood (LPL, CD36), intracellular FA transport (FABP3), intracellular FA activation of long- and short-chain FAs (ACSS1, ACSS2, ACSL1), de novo synthesis (ACACA), desaturation (SCD), triacyglycerol (TAG) synthesis (AGPAT6, GPAM, LPIN1), lipid droplet formation (PLIN2, BTN1A1, XDH), ketone body utilisation (BDH1, OXCT1), and transcription regulation (THRSP, PPARGC1A). In particular, intracellular de novo FA synthesis (ACSS2, ACACA, and FABP3) and TAG synthesis (GPAM, AGPAT6, and LPIN1), whose regulation might be orchestrated as part of the gene network under the control of SERBF1 in the milk fat synthesis process, were more activated compared to levels in dairy cows. However, the genes involved in lipid droplet formation (PLIN2, XDH, and BTN1A1) were expressed at lower levels compared to those in dairy cows, where these genes are mainly controlled by the PPARG regulator.

An experiment was conducted to study the seasonal changes in the fatty acid profile of milk from yaks (Bos grunniens) when kept at altitudes of 3000 m above sea level (a.s.l.) and higher. Data and samples were collected in summer (July), autumn (September), winter (November) and spring (March) from ten lactating yaks (four in spring). The yaks grazed pastures adjacent to the farm building throughout the year. In spring only they received 0·6 kg crop by-products per day (dry matter basis). Fresh alpine grasses, available in summer and autumn, showed high concentrations of α-linolenic acid (46–51 g/100 g lipids) compared with the dry, yellow vegetation of winter and spring (16 g/100 g lipids). In autumn and summer, the milk fat had higher concentrations of polyunsaturated fatty acids than in winter. These polyunsaturated fatty acids were comprised of vaccenic acid, rumenic acid and α-linolenic acid, which are all considered beneficial to human health. The rare fatty acid, γ-linolenic acid, was also detected in yak milk, especially in the milk obtained in spring. The results suggest that yak milk, which is the most important basic food of the Tibetan herders, has the most favourable fatty acid profile when yaks grazed green pasture, which also corresponds to the period of highest milk production.

We investigated and quantified the factors which may affect the prevalence of cystic echinococcosis caused by Echinococcus granulosus in Rangtang County using a multidisciplinary approach. From a previously performed field survey, epidemiological data were linked with environmental data. Altitude and land surface temperature were extracted from remote-sensing images. Cumulative logistic regression models were used to identify and quantify the potential risk factors. The multiple regression models confirmed that yaks (χ 2 = 4·0447, P = 0·0443), dogs (χ 2 = 8·3455, P = 0·0039) and altitude (χ2 = 7·6223, P = 0·0058) were positively correlated with the prevalence of cystic echinococcosis, while land surface temperature may have a negative association. The findings showed that dogs and yaks play the most important role in the transmission of cystic echinococcosis, while altitude and land surface temperature may also be involved in the transmission.

Interspecies somatic cell nuclear transfer (iSCNT), a powerful tool in basic scientific research, has been used widely to increase and preserve the population of endangered species. Yak (Bos grunniens) is one of these species. Development to term of interspecies cloned yak embryos has not been achieved, possibly due to abnormal epigenetic reprogramming. Previous studies have demonstrated that treatment of intraspecies cloned embryos with (NaBu) significantly improves nuclear–cytoplasmic reprogramming and viability in vitro. Therefore, in this study, we evaluated the effect of optimal NaBu concentration and exposure time on preimplantation development of yak iSCNT embryos and on the expression patterns of developmentally important genes. The results showed that 8-cell rate, blastocyst formation rate and total cell number increased significantly compared with their untreated counterparts when yak iSCNT embryos were treated with 5 nM NaBu for 12 h after activation, but that the 2-cell stage embryo rate was not significantly different. The treatment of NaBu also increased significantly the expression levels of Oct-4 and decreased the expression levels of HDAC-2, Dnmt-1 and IGF-1; the expression patterns of these genes were more similar to that of their bovine–yak in vitro fertilization (BY-IVF) counterparts. The results described above indicated that NaBu treatment improved developmental competence in vitro and ‘corrected’ the gene expression patterns of yak iSCNT embryos.

Domestic yak (Bos grunniens) metallothionein-I (MT-I) and metallothionein-III (MT-III) cDNA 3′-end full-length sequences (331 and 378 bp; GenBank accession MT-I no. AY758557, MT-III no. DQ492300) from the total RNA of liver and brain tissues were amplified and cloned by reverse transcription polymerase chain reaction (RT-PCR) and 3′-rapid amplification of cDNA ends (RACE) using the primers YMT-I SP and M13 primers M4, and YMT-III SP and M13 primers M4, respectively, which included the MT-I (183 bp) coding sequence, MT-III (207 bp) cDNA coding sequences, and also the tailing signal AATAAA and Poly(A) at the 3′-ends of the MT-I and MT-III, respectively. The analyses showed that the yak cDNA sequence coding for MT-I protein was composed of 61 amino acids, including 20 cysteines having a conserved tripeptide structure, for example C–X–C, C–C–X–C–C, C–X–X–C and so on. The yak cDNA sequence coding for MT-III protein was composed of 68 amino acids, including 19 cysteines having both MT-III-specific conserved tripeptides and the same conserved tripeptides as MT-I, including T, CPCP, GEGAEA and so on. They were all comparatively conservative in molecular evolution. These structures indicated that MT-III had the same physiological functions regarding heavy metals (detoxification and so on) as MT-I, and also different specific functions from MT-I, including inhibition of neuron growth.

The purpose of this study was to evaluate the effects of donor sex, treatments of cell cycle synchronization and donor nuclei obtained from fresh or frozen–thawed conditions on developmental competence of yak–bovine interspecies nuclear transfer embryos. Bovine (Bos taurus) oocytes were used as recipients and yak (Bos grunniens) ear fibroblast cells were used as donors. Results indicated that the development rate of male blastocysts was higher than that of female (56.6% versus 39.5%, P<0.05), whereas cleavage and total cell number showed no difference between the two groups. No significant difference was observed in the development and quality of blastocysts with donor cells treated by serum starvation or contact inhibition, and there was no significant difference in embryo development with fresh or frozen–thawed donor cells, whereas the cleavage rate in the group of frozen–thawed cells was significantly lower than that of the fresh cell group (54.5% versus 78.2%, P<0.05). The results demonstrated that donor sex could impact the developmental competence of yak–bovine interspecies nuclear transfer embryos, whereas different treatments of cell cycle synchronization and freezing had little influence.

It is necessary to fully understand the economic conditions of local herders in order to find solutions to the conflicts between wildlife conservation and livestock rearing in remote areas of low-income countries. In the Kanchenjunga Conservation Area (KCA), Nepal, livestock depredation by snow leopards impacts on yak herders' livelihoods. Retaliatory killings of snow leopard by the herders have been reported and the concerned authorities recently initiated snow leopard conservation programmes. In 2001, interviews with the yak herders who used the pastures in the Ghunsa valley in the preceding year collected data on the incidence of livestock death caused by snow leopards. The annual net cash income of the yak herders was estimated by obtaining baseline values of sales and expenditure per livestock head through field measurement of dairy products and interviews with a sample of herders. As yet, the average annual damage does not appear to have adversely affected fundamental livelihoods in households with an average herd size (36.6 head). However, in the worst scenario of livestock depredation, households with medium or small-sized herds (<40 head) might risk their living conditions becoming unsustainable or having to withdraw from yak pastoralism. A supplementary interview showed that the majority of the herders, except those who took completely neutral attitudes towards the regional conservation and development programme, had negative views of the snow leopard conservation policy. For the snow leopard conservation programme in the KCA to be a success, there must be a system to compensate the herders' households for livestock damage.

Thirty growing yaks Bos grunniens or Poephagus grunniens, 1·0–3·5 years and 50–230kg, from their native altitudes (3000–4000m), were used to study the basal metabolism in this species and to evaluate the effects of high altitude and season on the energy metabolism. Fasting heat production (FHP) was measured at altitudes of 2260, 3250 and 4270m on the Tibetan plateau in both the summer and the winter, after a 90d adaptation period at each experimental site. Gas exchanges of the whole animals were determined continuously for 3d (4–5 times per d, 10–12 min each time) after a 96 h starvation period, using closed-circuit respiratory masks. Increasing altitude at similar ambient temperature (Ta) did not affect (P>0·10) FHP in the summer, but decreased (P<0·05) it at different Ta in the winter. However, the decrease of FHP in the winter was mainly due to the decrease of Ta instead of the increase of altitude. In the summer, the respiratory rate, heart rate and body temperature were unaffected by altitude, except for a decrease (P<0·05) in body temperature at 4270m; in the winter, they were decreased (P<0·05) by increasing altitude. In both seasons, the RER was decreased (P<0·05) by increasing altitude. At all altitudes for all groups, the daily FHP was higher (P<0·05) in the summer (Ta 6–24°C) than in the winter (Ta 0 to -30°C), and the Ta-corrected FHP averaged on 920 kJ/kg body weight0·52 at Ta 8–14°C and on 704 kJ/kg body weight0·52 at Ta -15°C respectively. We conclude that in the yak high altitude has no effect on the energy metabolism, whereas the cold ambient temperature has a significant depressing effect. The results confirm that the yak has an excellent adaptation to both high altitude and extremely cold environments.

The Ladakh plateau is an arctic desert in the Western Himalaya having an annual rainfall of 8 to 9 cm. Extreme environmental conditions and persistent hypoxia affect the human and livestock populations. Limited grazing resources are the other limiting factor. Despite these hardships, a great variety of livestock species are available in Ladakh. The yak is the most important species for highlanders providing them subsistence in terms of milk, meat, fibres, skin and hides. It is an excellent pack and transport animal in snow bound passes. Male Yak hybrids with local cattle are excellent bullocks under the hypoxic environment. The small population of local hill cattle is endangered due to large scale crossbreeding with yaks and exotic cattle. The majority of goatsoin Ladakh are Pashmmina type. The Changthangi is an important goat breed. Local small sized sheep yield fine carpet type wool and resemble the Tibetan sheep. Zaniskari horses are in danger of extinction. Other horses and ponies available in Ladakh appeared similar to the Tibetan ponies. The local donkeys have a large bulging head with long hair. Only 54 Bacterin camels are left in the region. Poultry and pigs are not popular in Ladakh. Almost all livestock species have long body hair to survive in extreme climatic conditions.