First report of Ikeda genotype of Theileria orientalis in Mithun (Bos frontalis) from northeastern hilly region of India.

Oriental theileriosis caused by Theileria orientalis, previously considered a benign disease, is posing a significant threat to the livestock industry across the globe. To elucidate the prevalence of Theileria orientalis in ticks and their host, the Mithun, a comprehensive study was undertaken in the two northeastern states of India, viz. Nagaland and Arunachal Pradesh. A total of 340 of Rhipicephalus microplus ticks and 25 Ambylomma sp. ticks were screened for the presence of Theileria orientalis through PCR. Among the R. microplus ticks examined, 25 of them tested positive for T. orientalis infection whereas none of the Amblyomma ticks was positive. Additionally, a total of 275 blood samples were collected from Mithun from Arunachal and Nagaland and 31 animals were found to be positive for T. orientalis infection. Notably, six positive cases were identified in Porba (Phek district), six in Tening, and one in Bamsiakilwa village (Peren district) of Nagaland. Moreover, out of the 41 animals examined at Medziphema farms, Nagaland, 18 were found to be positive for T. orientalis infection. Moreover, the phylogenetic investigation has unveiled the presence of the highly pathogenic Type 2 (Ikeda) T. orientalis genotype in Mithun, supported by a strong bootstrap value of 100%. This study marks the initial documentation of oriental theileriosis in mithun. It underscores the need for vigilant monitoring and active surveillance of mithun populations in the northeastern states of India. Timely treatment of infected animals is imperative to avert economic losses for the farmers.

Muscle transcriptome signature and gene regulatory network analysis in two divergent lines of a hilly bovine species Mithun (Bos frontalis).

A massive bovine, Bos frontalis, also known as Mithun or Gayal, found at higher altitude is very promising meat and milk animal. For candidate gene and marker discovery, RNA-seq data was generated from longissimus dorsi muscle tissues with Illumina-HiSeq. Such markers can be used in future for genetic gain of traits like feed conversion efficiency (FCE) and average daily gain (ADG). Analysis revealed 297differentially expressed genes (DEGs) having 173 up and 124 down-regulated unigenes. Extensive conservation was found in genic region while comparing with Bos taurus. Analysis revealed 57 pathways having 112 enzymes, 72 transcriptional factors and cofactors, 212 miRNAs regulating 71 DEGs, 25,855 SSRs, mithun-specific 104,822 variants and 7288 indels, gene regulatory network (GRN) having 24 hub-genes and transcriptional factors regulating cell proliferation, immune tolerance and myogenesis. This is first report of muscle transcriptome depicting candidate genes with GRN controlling FCE and ADG. Reported putative molecular markers, candidate genes and hub proteins can be valuable genomic resources for association studies in genetic improvement programme.

Whole genome sequence and de novo assembly revealed genomic architecture of Indian Mithun (Bos frontalis).

BACKGROUND: Mithun (Bos frontalis), also called gayal, is an endangered bovine species, under the tribe bovini with 2n = 58 XX chromosome complements and reared under the tropical rain forests region of India, China, Myanmar, Bhutan and Bangladesh. However, the origin of this species is still disputed and information on its genomic architecture is scanty so far. We trust that availability of its whole genome sequence data and assembly will greatly solve this problem and help to generate many information including phylogenetic status of mithun. Recently, the first genome assembly of gayal, mithun of Chinese origin, was published. However, an improved reference genome assembly would still benefit in understanding genetic variation in mithun populations reared under diverse geographical locations and for building a superior consensus assembly. We, therefore, performed deep sequencing of the genome of an adult female mithun from India, assembled and annotated its genome and performed extensive bioinformatic analyses to produce a superior de novo genome assembly of mithun. RESULTS: We generated ≈300 Gigabyte (Gb) raw reads from whole-genome deep sequencing platforms and assembled the sequence data using a hybrid assembly strategy to create a high quality de novo assembly of mithun with 96% recovered as per BUSCO analysis. The final genome assembly has a total length of 3.0 Gb, contains 5,015 scaffolds with an N50 value of 1 Mb. Repeat sequences constitute around 43.66% of the assembly. The genomic alignments between mithun to cattle showed that their genomes, as expected, are highly conserved. Gene annotation identified 28,044 protein-coding genes presented in mithun genome. The gene orthologous groups of mithun showed a high degree of similarity in comparison with other species, while fewer mithun specific coding sequences were found compared to those in cattle. CONCLUSION: Here we presented the first de novo draft genome assembly of Indian mithun having better coverage, less fragmented, better annotated, and constitutes a reasonably complete assembly compared to the previously published gayal genome. This comprehensive assembly unravelled the genomic architecture of mithun to a great extent and will provide a reference genome assembly to research community to elucidate the evolutionary history of mithun across its distinct geographical locations.

High-density Genotyping reveals Genomic Characterization, Population Structure and Genetic Diversity of Indian Mithun (Bos frontalis).

The current study aimed at genomic characterization and improved understanding of genetic diversity of two Indian mithun populations (both farm, 48 animals and field, 24 animals) using genome wide genotype data generated with Illumina BovineHD BeadChip. Eight additional populations of taurine cattle (Holstein and NDama), indicine cattle (Gir) and other evolutionarily closely related species (Bali cattle, Yak, Bison, Gaur and wild buffalo) were also included in this analysis (N = 137) for comparative purposes. Our results show that the genetic background of mithun populations was uniform with few possible signs of indicine admixture. In general, observed and expected heterozygosities were quite similar in these two populations. We also observed increased frequencies of small-sized runs of homozygosity (ROH) in the farm population compared to field mithuns. On the other hand, longer ROH were more frequent in field mithuns, which suggests recent founder effects and subsequent genetic drift due to close breeding in farmer herds. This represents the first study providing genetic evidence about the population structure and genomic diversity of Indian mithun. The information generated will be utilized for devising suitable breeding and conservation programme for mithun, an endangered bovine species in India.

Effect of Addition of Taurine on the Liquid Storage (5°C) of Mithun (Bos frontalis) Semen.

The present study was undertaken to assess the effect of taurine on sperm motility, viability, total sperm abnormalities, acrosomal and plasma membrane integrity, enzymatic profiles such as reduced glutathione (GSH), glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT), and biochemical profiles such as cholesterol efflux and malondialdehyde (MDA) production. A total of 50 ejaculates were collected twice a week from 8 mithun bulls, and semen was split into 4 equal aliquots and diluted with the TEYC extender. Group 1: semen was without additives (control); groups 2 to 4: semen was diluted with 25 mM, 50 mM, and 100 mM of taurine, respectively. Seminal parameters and enzymatic and biochemical profiles were assessed at 5°C. Inclusion of taurine into diluent resulted in significant (P < 0.05) decreases in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities after liquid storage compared with the control group. Additionally, taurine at 50 mM has significant improvement in quality of mithun semen than taurine at 25 or 100 mM stored in in vitro at 5°C. It was concluded that the possible protective effects of taurine on sperm parameters are from enhancing the function of antioxidant enzymes, preventing efflux of cholesterol from cell membranes and decreased MDA production.

Growth performance and nutrient utilization in male and female Mithun calves on green forage-based diet.

A comparative study of growth rate and feed intake and its utilization in male and female Mithun calves was carried out in hilly tracts of Nagaland. The animals of both sexes were fed fresh jungle grasses ad libitum and 2.0 kg of rice bran fortified with 1% mineral mixture and 1% salt daily. Growth rates of 369.50 and 305.67 g/day were observed in male and female calves, respectively. Sex-related differences in feed and nutrient intakes were statistically significant, whereas no significant difference was found in digestibility coefficients of nutrients between the groups as well as among the animals. Dry matter, digestible crude protein and total digestible nutrient intakes were recorded to be 107.81, 8.08 and 74.73 g/kgW0.75 per day, respectively, irrespective of sex. Feed conversion efficiency (FCE) and nutritive value of the ration were similar in both male and female calves. Results obtained from the study indicate that the growth rate and nutrient intakes were higher in male than female calves but the sex of the animal did not have any significant effect on nutrient utilization.