Oregano Essential Oil as a Natural Plant Additive Affects Growth Performance and Serum Antibody Levels by Regulating the Rumen Microbiota of Calves.

This experiment aimed to investigate whether supplementation of calves with different doses of oregano essential oil (OEO) could promote the development of the gastrointestinal tract and enhance the immune ability of calves by regulating the rumen microbiota. Twenty-four 70-day-old healthy and disease-free Holstein male calves were randomly divided into four groups, with the control group fed a basal diet, and the treatment group provided 4 g, 6 g, and 8 g of oregano essential oil per day in addition to the basal diet. After the 14-day pre-test, a 56-day formal test was conducted. At days 0 and 56 of the standard test period, calves were weighed, the average daily weight gain of calves during the test period was calculated, and serum samples were collected to measure the concentration of immunoglobulins (IgA, IgG, and IgM) in the serum; at day 56 of the formal test period, rumen fluid was collected from the calves, and 16SrRNA was sequenced to analyze changes in the rumen microbiota of the calves. The changes in the rumen microbiota of calves were analyzed by 16SrRNA sequencing. The results of the study showed that (1) OEO supplementation in calves significantly increased end weight and average daily gain (p < 0.05); (2) OEO supplementation in calves significantly increased serum concentrations of immunoglobulins IgA and IgM (p < 0.05); (3) OEO supplementation in calves significantly increased the abundance and diversity of rumen microbial organisms (p < 0.05); (4) OEO supplementation in calves significantly regulates the relative abundance of some species, and biomarkers with significant differences were screened by LEfSe analysis: g_Turicibacter, g_Romboutsia, f_Peptostreptococcaceae, f_Clostridiaceae, g_Clostridium_sensu_stricto_1, o_Clostridiales, g_unclassified_f_Synergistaceae, c_Coriobacteriia, o_Coriobacteriales, f_Atopobiaceae, g_Olsenella, p_Actinobacteriota, g_Defluviitaleaceae_UCG-011, f_Defluviitaleaceae, o_Corynebacteriales, g_Corynebacterium, f_Corynebacteriaceae, g_Shuttleworthia, f_Hungateiclostridiaceae, o_norank_c_Clostridia, g_Saccharofermentans, g_Streptococcus, f_Streptococcaceae, g_unclassified_o_Oscillospirales, and f_unclassified_o_Oscillospirales (p < 0.05, LDA ≥ 3); and (5) OEO supplementation in calves significantly enriched the metabolism of cofactors and vitamins pathway (p < 0.05). (6) Using Superman's correlation analysis, we screened unclassified_c_Clostridia, Shuttleworthia, and Christensenellaceae_R-7_group, three beneficial strains for calves. (7) Daily supplementation with 8g of OEO significantly affected rumen microbiota regulation in calves.

Verification of Key Target Molecules for Intramuscular Fat Deposition and Screening of SNP Sites in Sheep from Small-Tail Han Sheep Breed and Its Cross with Suffolk.

Intramuscular fat (IMF) is vital for meat tenderness and juiciness. This study aims to explore the IMF deposition mechanism and the related molecular markers in sheep. Two populations, Small-tail Han Sheep (STH) and STH × Suffolk (SFK) F1 (SFK × STH), were used as the research object. Histological staining techniques compared the differences in the longissimus dorsi muscle among populations. A combination of transcriptome sequencing and biological information analysis screened and identified IMF-related target genes. Further, sequencing technology was employed to detect SNP loci of target genes to evaluate their potential as genetic markers. Histological staining revealed that the muscle fiber gap in the SFK × STH F1 was larger and the IMF content was higher. Transcriptome analysis revealed that PIK3R1 and PPARA were candidate genes. Histological experiments revealed that the expressions of PIK3R1 mRNA and PPARA mRNA were lower in SFK × STH F1 compared with the STH. Meanwhile, PIK3R1 and PPARA proteins were located in intramuscular adipocytes and co-located with the lipid metabolism marker molecule (FASN). SNP locus analysis revealed a mutation site in exon 7 of the PIK3R1 gene, which served as a potential genetic marker for IMF deposition. This study's findings will provide a new direction for meat quality breeding in sheep.

Metagenomic and metabolomic analysis of changes in intestinal contents of rainbow trout (Oncorhynchus mykiss) infected with infectious hematopoietic necrosis virus at different culture water temperatures.

Infectious hematopoietic necrosis (IHN) is a major disease that limits the culture of rainbow trout. In practical production, it has been found that the temperature of the culture water is a crucial factor affecting its mortality. Currently, little is known about how temperature affects the immune response of rainbow trout gut microbiota and metabolites to IHNV. In this study, our main objective is to analyze the changes in gut microorganisms of rainbow trout (juvenile fish with a consistent genetic background) after 14 days of infection with IHNV (5 × 105 pfu/fish) at 12-13°C (C: injected with saline, A: injected with IHNV) and 16-17°C (D: injected with saline, B: injected with IHNV) using metagenomic and metabolomic analyses, and to screen for probiotics that are effective against IHNV. The results showed that infection with IHNV at 12-13°C caused Eukaryote loss. Compared to Group C, Group A showed a significant increase in harmful pathogens, such as Yersiniaceae, and a significant alteration of 4,087 gut metabolites. Compared to group D, group B showed a significant increase in the abundance of Streptococcaceae and Lactococcus lactis, along with significant changes in 4,259 intestinal metabolites. Compared with their respective groups, the levels of two immune-related metabolites, 1-Octadecanoyl-glycero-3-phosphoethanolamine and L-Glutamate, were significantly upregulated in groups A and B. Compared to group B, Group A showed significantly higher pathogenic bacteria including Aeromonas, Pseudomonas, and Yersiniaceae, while group B showed a significant increase in Streptococcaceae and Lactococcus lactis. Additionally, there were 4,018 significantly different metabolites between the two groups. Interestingly, 1-Octadecanoyl-sn-glycero-3-phosphoethanolamine and L-Glutamate were significantly higher in group A than in group B. Some of the different metabolites in C vs. A are correlated with Fomitopsis pinicola, while in D vs. B they were correlated with Lactococcus raffinolactis, and in A vs. B they were correlated with Hypsizygus marmoreus. This study exposed how rainbow trout gut microbiota and metabolites respond to IHNV at different temperatures, and screens beneficial bacteria with potential resistance to IHN, providing new insights and scientific basis for the prevention and treatment of IHN.

Effects of Essential Oil and/or Encapsulated Butyrate on Fecal Microflora in Neonatal Holstein Calves.

This study was conducted to investigate the effects of feeding oregano essential oil, butyrate, and its mixture on the intestinal microbial diversity of calves. A completely randomized experimental design was used. Sixty-four healthy neonatal Holstein female calves with birth weight ≥ 35 kg were randomly divided into one control and three treatments (16 calves per group). The control group was fed normally, and the treatment group was fed oregano essential oil, butyrate, and their mixture, respectively. The experiment lasted for 70 days, and the lactation period lasted for 56 days. On days 55 and 70, rectal fecal samples from five calves were collected from each group for 16S rRNA amplification and sequencing. The results showed as follows: (1) the three treatments had no significant effects on the intestinal microbial community diversity, community uniformity, and community pedigree diversity of calves (p > 0.05). (2) At the phylum level, Firmicutes, Bacteroidota, Spriochatetota, Actinobacteriota, Firmicutes, and Bacteroidota gates of the main bacteria were detected in feces. (3) At the genus level, the top ten species with relative abundance detected are: norank_ F_Muribaaculaceae, Ruminococcus, unclassified_ F_ Lachnospiraceae, UCG-005, Prevotelaceae_NK3B31_Group, Prevotella, Bacteroides, Rikenellaceae_RC9_Gut_Group, and Faecalibacterium, Alloprevotella. (4) LEfSe analysis results show that the species with significant differences in the control group were f__Lachnospiraceae, o__Lachnospirales, o__Coriobacteriales, and c__Coriobacteriia, g__Megasphaera; in the essential oil group were g__Lachnospiraceae_AC2044_group, o__Izemoplasmatales, g__norank_f__norank_o__Izemoplasmatales, and f__norank_o__Izemoplasmatales; in the sodium butyrate group were g__Lachnospiraceae_NK4A136_group, and g__Sharpea, g__Fournierella; in the mixed group were g__Flavonifractor, and g__UBA1819. (5) The functional prediction analysis of calf gut microbes, found on the KEGG pathway2, shows that essential oil significantly improved membrane transport, Sodium butyrate inhibits lipid metabolism and improves the body's resistance to disease. (p < 0.05). (6) The effects of each treatment on the intestinal microbial structure of calves did not last for 14 days after the treatment was stopped. In conclusion, the addition of oregano essential oil, butyrate, and its mixtures to milk fed to calves can modulate the microbial structure, and it is recommended that oregano essential oil and butyrate be used separately, as a mixture of the two can increase the rate of diarrhea in calves.

Genome-wide single-nucleotide polymorphism data and mitochondrial hypervariable region 1 nucleotide sequence reveal the origin of the Akhal-Teke horse.

OBJECTIVE: The study investigated the origin of the Akhal-Teke horse using genome-wide single-nucleotide polymorphism (SNP) data and mitochondrial hypervariable region 1 (HVR-1) nucleotide sequences. METHODS: Genome-wide SNP data from 22 breeds (481 horses) and mitochondrial HVR-1 sequences from 24 breeds (544 sequences) worldwide to examine the origin of the Akhal- Teke horse. The data were analyzed using principal component analysis, linkage disequilibrium analysis, neighbor-joining dendrograms, and ancestry inference to determine the population relationships, ancestral source, genetic structure, and relationships with other varieties. RESULTS: A close genetic relationship between the Akhal-Teke horse and horses from the Middle East was found. Analysis of mitochondrial HVR-1 sequences showed that there were no shared haplotypes between the Akhal-Teke and Tarpan horses, and the mitochondrial data indicated that the Akhal-Teke horse has not historically expanded its group. Ancestral inference suggested that Arabian and Caspian horses were the likely ancestors of the Akhal- Teke horse. CONCLUSION: The Akhal-Teke horse originated in the Middle East.

Screening of Genes Related to Growth, Development and Meat Quality of Sahan Crossbred F1 Sheep Based on RNA-Seq Technology.

This study aimed to identify genes related to sheep growth, development and meat quality. Small-tailed Han sheep (STH), and small-tailed Han sheep and Suffolk crossbred F1 (STH×SFK), were selected to determine the growth performance, slaughter performance, and meat quality. The longissimus dorsi muscle was selected for transcriptome sequencing, and the target gene was screened based on bioinformatics analysis; real-time fluorescent quantitative PCR (RT-PCR) and western blotting (WB) were conducted to verify the target gene. Locations of genes in tissues were confirmed via immunofluorescence. The results showed that the pre-slaughter live weight, bust circumference, slaughter performance, and marbling score of the STH×SFK population were significantly higher than those of the STH population (P < 0.01). Sequencing results showed that 560 differentially expressed genes (DEGs) were identified in the STH×SFK population, of which 377 exhibited up-regulated and 183 exhibited down-regulated expression levels. GO annotation revealed that DEGs could be classified into 13 cell components, 10 molecular functions, and 22 biological processes. The KEGG enrichment analysis showed that DEGs were mainly enriched in the Rap1 signaling pathway, Ras signaling pathway, and other pathways related to growth and meat quality. Based on the GO and KEGG analyses, four candidate genes related to sheep growth and meat quality, namely myostain (MSTN), interferon-related developmental regulator 1 (IFRD1), peroxisome proliferator activator receptor delta (PPARD), and myosin light chain 2 (MLC2 or MYL2), were screened. The expression levels of genes and proteins were verified via RT-PCR and WB, and the results were consistent with the trend of transcriptome sequencing. Immunofluorescence results showed that IFRD1 was expressed in the cytoplasm and nucleus, and MYL2 was expressed in the cytoplasm. This study revealed the mechanism of gene regulation of sheep growth and development at the molecular level and provided a theoretical basis for studying sheep genetics and breeding.

Exploring effects of different male parent crossings on sheep muscles and related regulatory genes using mRNA-Seq.

OBJECTIVE: With improvements in living standards and increase in global population, the demand for meat products has been increasing; improved meat production from livestock could effectively meet this demand. In this study, we examined the differences in the muscle traits of different male crossbred sheep and attempted to identify key genes that regulate these traits. METHODS: Dubo sheep×small-tailed Han sheep (DP×STH) and Suffolk×small-tailed Han sheep (SFK×STH) were selected to determine meat quality and production performance by Masson staining. Transcriptome sequencing and bioinformatic analysis were performed to identify differentially expressed genes (DEGs) related to meat quality. The presence of DEGs was confirmed by real-time polymerase chain reaction. RESULTS: The production performance of SFK×STH sheep was better than that of DP×STH sheep, but the meat quality of DP×STH sheep was better than that of SFK×STH sheep. The muscle fiber diameter of DP×STH sheep was smaller than that of SFK×STH sheep. Twenty-two DEGs were identified. Among them, four gene ontology terms were related to muscle traits, and three DEGs were related to muscle or muscle fibers. There were no significant differences in the number of single nucleotide mutations and mutation sites in the different male parent cross combinations. CONCLUSION: This study provides genetic resources for future sheep muscle development and cross-breeding research.

Excavation of Genes Related to the Mining of Growth, Development, and Meat Quality of Two Crossbred Sheep Populations Based on Comparative Transcriptomes.

Crossbreeding can improve production performance and meat quality in sheep. The objective of this study was to look for genes related to sheep growth, development, and muscle. In this study, Dorper (DP) × Small Tailed Han (STH) sheep and Mongolia (MG) × Small-tailed Han (STH) sheep were used to estimate the productive performance and meat quality in a crossbreed. Subsequently, transcriptome analysis and bioinformatic analysis were performed on the Longissimus dorsi muscles of DP × STH and MG × STH sheep to identify differentially expressed genes (DEGs) related to growth, development, and meat quality. The presence of DEGs was confirmed by real-time PCR (qPCR). Productive performance and meat quality of the DP × STH sheep were better than the MG × STH sheep. Compared to DP × STH, a total of 1445 DEGs were identified in MG × STH sheep (1026 DEG were up-regulated and 419 DEG were down-regulated). Of these, 38 DEGs were related to growth, 161 to development, and 43 to muscle. In addition, 13 co-expressed genes (FGFRL1, SIX1, PLCB1, CRYAB, MYL2, ADIPOQ, GPX1, PPARD, GPC1, CDC42, LOC101106246, IGF1, and LARGE) were identified. The expression of DEGs was consistent with the comparative transcriptome analysis. This work provides genetics resources for future research on muscle development in sheep.

Fatty Acid Profile of Muscles from Crossbred Angus-Simmental, Wagyu-Simmental, and Chinese Simmental Cattles.

This study assessed breed differences in fatty acid composition and meat quality of Longissimus thoracis et lumborum (LTL) and semitendinosus (SE) of Angus× Chinese Simmental (AS), Wagyu×Chinese Simmental (WS), and Chinese Simmental (CS). CS (n=9), AS (n=9) and WS (n=9) were randomly selected from a herd of 80 bulls which were fed and managed under similar conditions. Fatty acid profile and meat quality parameters were analyzed in duplicate. Significant breed difference was observed in fatty acid and meat quality profiles. AS exhibited significantly (p<0.05) lower C16:0 and higher C18:1n9c compared with CS. AS breed also had a tendency (p<0.10) to lower total saturated fatty acid (SFA), improve C18:3n3 and total unsaturated fatty acid (UFA) compared with CS. Crossbreed of AS and WS had significantly (p<0.05) improved the lightness, redness, and yellowness of muscles, and lowered cooking loss, pressing loss, and shear force compared with CS. These results indicated that fatty acid composition and meat quality generally differed among breeds, although the differences were not always similar in different tissues. Fatty acid composition, meat color, water holding capacity, and tenderness favored AS over CS. Thus, Angus cattle might be used to improve fatty acid and meat quality profiles of CS, and AS might contain better nutritive value, organoleptic properties, and flavor, and could be potentially developed as an ideal commercial crossbreed.

Comparative analysis of Longissimus dorsi tissue from two sheep groups identifies differentially expressed genes related to growth, development and meat quality.

From a genetic perspective, the advantages of crossbreeding in sheep are unclear. In the present study, a comparative transcriptomic analysis was performed using Longissimus dorsi tissues from two sheep groups in order to identify differentially expressed genes (DEGs) related to growth, development and meat quality. Compared to Small Tail Han sheep, a total of 874 DEGs were identified in the crossbred sheep. Among these DEGs, 30, 116 and 32 DEGs were related to growth, development and meat quality, respectively. Seven DEGs highlighted by functional analysis as playing crucial roles in growth, development and meat quality were validated by the gene-act-network and co-expression-network. The expression levels of DEG mRNAs and proteins were further confirmed using RT-qPCR and western blot analyses. The results were consistent with the comparative transcriptome data. The data from this transcriptomic analysis will help to understand genetic heterosis and molecular-assisted breeding in sheep.

Comparative Transcriptome Analysis Identifying the Different Molecular Genetic Markers Related to Production Performance and Meat Quality in Longissimus Dorsi Tissues of MG × STH and STH Sheep.

Crossbred sheep have many prominent traits, such as excellent production performance and high-quality meat, when compared to local sheep breeds. However, the genetic molecular markers related to these characteristics remain unclear. The crossbred MG × STH (small-tailed Han sheep (STH) × Mongolian sheep (MG)) breed and the STH breed were selected to measure production performance and meat quality. We used 14 indexes of production performance and meat quality, which in the MG × STH population showed significant differences compared to the STH breed. Subsequently, the longissimusdorsi from the two sheep were subjected to comparative transcriptomic analyses to identify differentially expressed genes (DEGs) related to production performance and meat quality. A total of 874 DEGs were identified between the two sheep groups. A total of 110 unique DEGs related to sheep production performance and meat quality were selected as the candidate DEGs. We found 6 production-performance-related and 30 meat-quality-related DEGs through a correlation analysis, including SPARC, ACVRL1, FNDC5 and FREM1. The expression levels of 11 DEGs were validated by real-time PCR, and the results were in accordance with the results of the comparative transcriptomic and correlation analyses. These results will assist in understanding sheep heterosis and molecular marker-assisted selection.

Calf starter containing a blend of essential oils and prebiotics affects the growth performance of Holstein calves.

Essential oils extracted from specific plants can exhibit antimicrobial properties that make them potential antibiotic alternatives. The objective was to evaluate an essential oil and prebiotic combination (EOC) on the growth, development, and health status of growing neonatal calves. Forty Holstein newborn calves were blocked by birth date and alternately assigned to 1 of 2 treatments. Treatments were a pelleted calf starter (CS) without (control) or with EOC at 44.1 ppm. Calves were fed the experimental CS for ad libitum consumption from 3 d of age through the end of the 70-d experiment. Calves were fed 2 L of whole milk twice daily to 10 d of age, then 3 L twice daily through d 35, and then fed 3 L once daily with abrupt weaning occurring after 42 d of age. The average daily gain (0.78 and 0.87 kg/d for control and EOC, respectively) was greater for calves fed EOC compared with calves fed the control. Calves fed EOC demonstrated greater dry matter intake (1.63 and 1.74 kg/d) compared with calves fed the control. Feed conversion ratio (0.62 and 0.65 kg of gain/kg of dry matter intake) was greater for calves fed EOC compared with calves fed the control. At 70 d of age, calves fed EOC demonstrated increased body frame measurements (hip height, body length, heart girth, abdominal girth, and pastern) compared with calves fed the control. The incidence of scours score 4 (mild diarrhea) was 3.5 incidences lower for calves fed EOC compared with calves fed the control, whereas the incidence of scours score 5 (severe diarrhea) for calves fed EOC was 0.5 incidences lower than calves fed the control. Blood concentrations of IgG and IgM on d 14, IgA on d 28, and total serum protein on d 42 were all greater for calves fed EOC compared with calves fed the control. The blood volatile fatty acid concentrations were greater for calves fed EOC compared with calves fed the control, which indicated enhanced ruminal development. Total-tract digestibility of dry matter, crude protein, acid detergent fiber, neutral detergent fiber, starch, and many minerals were increased for calves fed EOC compared with calves fed the control. The inclusion of an EOC blend into a CS demonstrates promising benefits for enhancing calf growth, ruminal development, gut health, nutrient digestibility, and immunity. The use of an EOC blend can be a plausible alternative to feeding subtherapeutic antibiotics for improving calf performance, health, and immunity.

Comprehensive Analysis of MicroRNA⁻Messenger RNA from White Yak Testis Reveals the Differentially Expressed Molecules Involved in Development and Reproduction.

Testis development is a vital and tightly regulated process in mammals. Understanding the biological mechanisms underlying testis development will benefit the animal reproduction industry. Expression changes in microRNA and messenger RNA in response to dynamic regulation effects have been associated with this process. However, very little is known about the roles of these molecules in yak development. Using whole-genome small RNA and messenger RNA sequencing, we performed a comprehensive analysis of the microRNA⁻messenger RNA interaction network expression in the testicles of Tianzhu white yaks during three developmental stages. Using Short Time-series Expression Miner analysis we identified 589 differentially expressed microRNAs (DERs) and 3383 differentially expressed messenger RNAs (DEGs) in the three age groups. A total of 93 unique DEGs are primarily involved in reproduction and testis development. Subsequently, four integration networks were constructed according to the DEGs and DERs in three biological processes. Nineteen DEGs were potentially regulated by 60 DERs, of which miR-574 and target gene AURKA played a crucial role in yak testis development and reproduction. The results of this study provide a basis for further exploration of the microRNA⁻messenger RNA interactions in testis development and reproduction and aid in uncovering the molecular mechanisms of spermatogenesis in male mammals.

Mutations in MC1R gene determine black coat color phenotype in Chinese sheep.

The melanocortin receptor 1 (MC1R) plays a central role in regulation of animal coat color formation. In this study, we sequenced the complete coding region and parts of the 5'- and 3'-untranslated regions of the MC1R gene in Chinese sheep with completely white (Large-tailed Han sheep), black (Minxian Black-fur sheep), and brown coat colors (Kazakh Fat-Rumped sheep). The results showed five single nucleotide polymorphisms (SNPs): two non-synonymous mutations previously associated with coat color (c.218 T>A, p.73 Met>Lys. c.361 G>A, p.121 Asp>Asn) and three synonymous mutations (c.429 C>T, p.143 Tyr>Tyr; c.600 T>G, p.200 Leu>Leu. c.735 C>T, p.245 Ile>Ile). Meanwhile, all mutations were detected in Minxian Black-fur sheep. However, the two nonsynonymous mutation sites were not in all studied breeds (Large-tailed Han, Small-tailed Han, Gansu Alpine Merino, and China Merino breeds), all of which are in white coat. A single haplotype AATGT (haplotype3) was uniquely associated with black coat color in Minxian Black-fur breed (P = 9.72E - 72, chi-square test). The first and second A alleles in this haplotype 3 represent location at 218 and 361 positions, respectively. Our results suggest that the mutations of MC1R gene are associated with black coat color phenotype in Chinese sheep.

[Origin and genetic diversity of Mongolian and Chinese sheep using mitochondrial DNA D-loop sequences].

To determine the origin and gene diversity of the Chinese and Mongolian domestic sheep, a partial fragment of mitochondrial DNA D-loop was sequenced for total number of 314 individuals from nine Chinese sheep populations and 11 Mongolian sheep populations. The results show no difference in nucleotide composition between Chinese and Mongolian sheep mtDNA D-loop sequences. However, more variables were identified in Mongolian sheep (26.85% of the sites) than that in Chinese sheep (24.22%). In China, mtDNA haplotype diversity was the highest in Qinghai Tibetan sheep, followed then by Gansu Tibetan sheep, Gansu Alpine Merino, Qinghai Merino, Gannan Tibetan sheep, Small-tailed Han sheep, Tan sheep, Hu sheep and Minxian Black Fur sheep. In Mongolian sheep, mtDNA haplotype diversity was the highest in Bayad and Baidrag populations and the lowest in the Gobi-Altai population. In general, Mongolian sheep have a richer genetic diversity than the Chinese ones with larger number of haplotypes (86.06% (142/165) versus 78.83% (108/137)), higher haplotype diversity (Hd; 0.976 versus 0.936), higher nucleotide diversity (Pi (pi); 0. 036 versus 0.034) and higher average number of nucleotide differences (k; 23.50 versus 22.48). Phylogenetic analysis of the 217 haplotypes identified in both Mongolian and Chinese sheep supported the same origin of their domestication with three distinct maternal lineages defined as major haplotypes A, B and C, of which haplotype A are the commonest in all Chinese sheep populations and in the majority of Mongolian sheep populations (9/11) with an average frequency of 58.73%, followed by haplotype B present in eight of Chinese population and in all Mongolian sheep populations with an average frequency of 24.68%, and haplotype C present in eight Chinese and in 10 Mongolian sheep populations with an average frequency of 16.59%. Further network analysis of the phylogenetic relationship of the 87 haplotypes identified from 91 sequences retrieved from GenBank together with the 217 haplotypes detected in this study reveals clearly four distinct lineages with the European mouflon (O. musimon) mixed into one of the lineages (haplotype B). There is no evidence of contribution of Argali sheep (O. ammon), O. vignei bochariensis and/or O. ammon nigrimontana to the maternal origin of both Mongolian and Chinese domestic sheep.