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The growth traits are economically important traits in sheep. Improving growth rates will increase the profitability of producers. The aim of this study was to identify alleles of CD8B (encoding T-cell surface glycoprotein CD8 beta chain) that are aberrantly expressed in different tissues and to assess the effects and associations of its different genotypes on weight and size traits in sheep. Using quantitative real-time reverse transcription PCR arrays, expression profiling of CD8B was performed in various organs and tissues. CD8B was ubiquitously expressed, with very high expression in the lung, spleen, lymph, duodenum, and liver. One intronic mutation (chr3:62,718,030 (Oar_rambouillet_v1.0, same below) G > A) was identified using pooled DNA sequencing. Subsequently, the variants (AA, AG, and GG) were genotyped using the KASPar® PCR single nucleotide polymorphism (SNP) genotyping system. The results of association analysis with body weight and body size traits in 1304 sheep showed that increases in multiple phenotypic traits correlated with the AA genotype (body weight, p < 0.05; body length, p < 0.05). Thus, SNP chr3:62,718,030 G > A is a promising molecular marker for marker-assisted selection in sheep breeding. [ABSTRACT FROM AUTHOR]

Dietary fiber supplementation can stimulate rumen development in lambs during the pre-weaning period. However, it is unclear whether different sources of fiber have varying effects on rumen development. This study aimed to investigate the molecular mechanism of rumen morphological and functional development based on non-forage or forage as a starter dietary fiber source. Twenty-four male Hu lambs with similar body weights (BW, 3.67 ± 0.08 kg) were selected and divided into two groups that received diets supplemented with either alfalfa hay (AH) or soybean hull (SH). At the age of 70 days, six lambs were slaughtered from each treatment group for rumen fermentation and morphological analyses. Three samples of the rumen tissue from the ventral sac were collected for transcriptomic analysis. The results identified 633 differentially expressed genes (DEGs), of which 210 were upregulated and 423 were downregulated in the SH group compared with those in the AH group. The upregulated DEGs were most enriched in the immune function and proteolysis pathways, whereas the downregulated DEGs were mainly involved in cell proliferation, apoptosis, and differentiation pathways. These findings indicated that non-forage as a starter dietary fiber source improved immune function and enhanced nitrogen utilization, whereas forage facilitated rumen morphological development. [ABSTRACT FROM AUTHOR]

Simple Summary: Early nutritional intervention in young animals can regulate physiological and growth performance at later stages. Previous studies have shown that early supplementation with starters can improve lamb growth performance; however, most of them focus on physiological changes in the short period before and after weaning of lambs, while reports on sustained effects on growth performance in later stages of lambs are scarce. This study investigated the sustained effects of early starter supplementation on the performance of Hu lambs during the pre-weaning, post-weaning, and fattening periods. The aim of this experiment was to study the sustained effects of early supplementation with a starter on the performance of lambs during the pre-weaning, post-weaning, and fattening periods. Sixty male Hu lambs (3.59 ± 0.05 kg) were randomly assigned to two (30 lambs per group) treatments. The lambs were fed milk replacer from three days of age. The early supplementation (ES) group was supplemented with a starter ration at seven days of age, the control (CON) group was supplemented at 21 days of age, and lambs in both groups were weaned from milk replacer at 28 days of age. Eight lambs from each group were randomly slaughtered at 98 days of age, and the remaining lambs were fed the same nutrient level of a fattening ration until slaughter at 200 days of age. The results showed that early supplementation with starter significantly improved average daily feed intake (ADFI) and average daily weight gain (ADG) in the pre-weaning period (7–28 days of age), and ADFI and slaughter performance (live weight before slaughter, carcass weight and dressing percentage) in the post-weaning period (29–98 days of age, p < 0.05). In addition, early supplementation with the starter had no significant effect (p > 0.05) on ADFI, ADG, and slaughter performance, but significantly decreased (p < 0.05) the feed-to-gain ratio (F/G) of lambs during the fattening period (99–200 days of age). In addition, early supplementation with starter increased the ratio of rumen and reticulum weight to total stomach weight in lambs (p < 0.05). In conclusion, early supplementation with starter can reduce feed costs and improve the performance of lambs, while the growth advantage produced by early supplementation had initially disappeared by 154 days of age. [ABSTRACT FROM AUTHOR]

• The content of fatty acids is influenced by the season. • The SLC27A6 is involved in fatty acid metabolism. • SLC27A6 has 7 SNPs significantly impacting at least one fatty acid phenotype. Fatty acids (FA) are an important factor affecting meat quality and human health, and the important role of the solute carrier family 27 member 6 (SLC27A6) in FA metabolism has been demonstrated in several species. However, the expression profile of the SLC27A6 in different tissues and the effect of its polymorphism on FA in sheep are currently unknown. This study aimed to explore the differences in FAs in the longissimus dorsi (LD) of 1,085 Hu sheep, the expression profile of SLC27A6 , and confirm the effect of single nucleotide polymorphisms (SNPs) on FA phenotypes. We found that many FA phenotypes differ significantly across different seasons, and winter promoted the deposition of polyunsaturated fatty acids (PUFA). The mRNA expression level of SLC27A6 in the lung was significantly higher than that in the heart, testis, and LD. A total of 16 SNPs were detected in the SLC27A6 , and 14 SNPs were successfully genotyped by improved multiplex ligase detection reaction (iMLDR) technology. Correlation analysis showed that 7 SNPs significantly affected at least one FA phenotype. Among them, SNP14 contributes to the selection of lamb with low saturated fatty acid content and high PUFA content. Combined genotypes also significantly affected a variety of beneficial FAs such as C18:3n3, C20:4n6, C22:6n3, and monounsaturated fatty acids. This study suggests that SLC27A6 plays an important role in FA metabolism and SNPs that are significantly associated with FA phenotype could be used as potential molecular markers for later targeted regulation of FA profiles in sheep. [ABSTRACT FROM AUTHOR]

The growth rate of sheep determines their productivity; the faster the growth rate, the higher the productivity. Sheep growth is usually measured by body weight, which is an important indicator of overall growth, and body size, which reflects body shape and structure. Therefore, in sheep production, the selection and improvement of growth traits is one of the important means to improve production efficiency. In this study, 857 males of Hu sheep were selected to record growth traits, including body weight, height, length, and chest, and cannon circumference at 80, 100, 120, 140, 160 and 180 days of age, as well as live weight before slaughter and carcass weight. Correlation analyses conducted between these growth traits at different stages were significantly correlated with both live weight before slaughter and carcass weight. And the strongest correlation was observed for body weight, while body height showed the weakest correlation. The myosin heavy chain 3 (MYH3) gene in the MYH gene family was selected as the target gene, and a synonymous mutation (MYH3 g.33834192A>G) in the exon 12 region of MYH3 was identified by polymerase chain reaction (PCR) and Sanger sequencing. Further association analysis using competitive allele-specific PCR (KASPar) technique showed that the MYH3 g.33834192A>G locus was significantly (P < 0.05) associated with growth traits (including body weight from 80 to 160 days of age, body height at 80 and 100 days of age, body length at 100 and 140 days of age, chest circumference at 80, 100, 160, and 180 days of age, cannon circumference at 80–140 days of age and carcass weight). Among them, the dominant genotype was GG. Meanwhile, the tissue expression results indicated that the highest level of MYH3 gene expression was found in muscle tissues. Therefore, this locus can be used as a candidate molecular genetic marker for improving growth traits in Hu sheep. • Correlation analysis yielded a significant positive correlation between body weight, body measurements, live weight before slaughter and carcass weight at each stage. • A synonymous mutation in the exon 12 region of the MYH3 gene of the Hu sheep was tested (MYH3 g.33834192 A>G). • This SNP locus was significantly (P < 0.05) associated with body weight, body height, body length, chest circumference, cannon circumference and carcass weight with dominant genotype GG. • RT-qRCR results showed that the expression of MYH3 gene in muscle tissue was significantly higher than that in the other 9 tissues (P < 0.05). [ABSTRACT FROM AUTHOR]

The purpose of this study was to investigate the effects of adding mannan-oligosaccharides (MOS) on the following parameters in sheep: digestibility and retention rate of nutrients, ruminal fermentation, immunity, and antioxidant capacity. Twelve healthy crossbred wethers (Suffolk ♂ × Small tail Han-yang ♀) with external ruminal fistula and similar body weights (28.04 ± 2.07 kg) were fed individually four treatments, three repeats of each treatment. The wethers diets were supplemental MOS at 0%, 1.2%, 1.6%, and 2.0%·kg-1 of basal diet (as fed basis). The experiment lasted 17 d, including 10 d of acclimation and 7 d of formal experimentation. The results showed that MOS did not influence the apparent digestibility and retention rate of nutrients, ruminal fermentation, and immunity or concentration of serum nitric oxide and activity of serum nitric oxide synthase (P ≥ 0.07). However, the apparent digestibility of neutral detergent fiber and acid detergent fiber at MOS supplementation rates of 1.6% and 2.0% both tended to be greater than the control group (P ≤ 0.103). There was also moderate evidence that MOS might increase the nitrogen retention rate (P = 0.082). MOS increased the antioxidant ability of sheep (P ≤ 0.018), especially at a dose of 1.6%: an increase in activity of total superoxide dismutase (P = 0.007), glutathione peroxidase (P = 0.018) and total antioxidant capacity (P < 0.001), and a decrease in concentration of malondialdehyde (P < 0.001) were found. The results indicated that in sheep MOS improved fiber digestion, N retention and some antioxidant abilities, but these effects may be too small to improve health and performance. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the effects of early starter feeding on ruminal pH regulation and the expression levels of genes involved in rumen volatile fatty acid (VFA) metabolism in lambs. A total of 78 male Hu lambs with similar born body weights were selected. Six lambs were slaughtered immediately after birth, and the remaining 72 were equally divided into a group fed by starter diet 1 at day 7 or a group fed by starter diet 1 at day 42. The lambs from both groups were weaned at day 56 and transacted to starter diet 2 at day 59. Six lambs from each group were randomly slaughtered at days 14, 28, 42, 56, 70 and 84, respectively. The results revealed that the ruminal VFA concentration in the early provision group was higher ( P < 0.05) than that in late provision group, while the rumen pH was not affected ( P > 0.05) by early or late starter feeding. Treatment-age interactions ( P < 0.05) and age alone ( P < 0.05) affected the expressions of NHE2, NHE3, DRA, HMGCL and MCT1. The expression of NHE2, DRA and HMGCL in early provision group were lower ( P < 0.05) than that in late provision group. However, the expression of MCT1 in early provision group was higher ( P < 0.05) than that in the late provision group. The findings of this study show that early starter feeding in lambs can increase the ruminal VFA concentration and affect the expression levels of genes involved in VFA transport and pH regulation in the ruminal epithelium. [ABSTRACT FROM AUTHOR]

Increased body growth and feed efficiency are important for breeding due to expensive feed costs. This study estimated the genetic parameters for two feed efficiency traits [the feed conversion ratio (FCR) and the residual feed intake (RFI)] and other important economic traits in male Hu sheep using 1642 male Hu lambs. The heritability of the RFI was estimated at 0.31 (±0.10)–0.54 (±0.1) and the heritability of the FCR was 0.05 (±0.07)–0.77 (±0.12). There was a negative genetic correlation between rib-eye muscle area traits and RFI. A positive genetic correlation was found between fat deposition traits and feed efficiency traits. The digestive tract weight decreased with the direction of RFI: however, the FCR showed the opposite: A low FCR was accompanied by a high digestive tract weight. The genetic correlation between feed efficiency (RFI and FCR) and rumen weight was negative. Heritability estimates for feed efficiency generate fluctuations with different experimental intervals. The weight of other traits should be reasonably allocated to avoid losses in production and feed efficiency in the selection of feed efficiency traits. The present study added to our understanding of the genetic parameters of sheep under the condition of house feeding. [ABSTRACT FROM AUTHOR]

Meat quality is becoming more important for sheep breeding programs. Meat quality is a complex trait affected by genetic and environmental factors. In the present study, an integrative analysis of the longissimus thoracis tissue transcriptome and proteome was conducted to identify genes, proteins, and pathways related to meat quality in sheep. The sheep breeds Hu and Dorper were considered. These breeds were compared for the differences in muscle fiber structure, chemical composition, and amino acid composition. In the Hu sheep vs. Dorper sheep comparison, 22 DEGs/DEPs showed the same mRNA and protein expression trends. These genes are associated with lipid transport, lipid metabolism, and muscular system development. Moreover, some pathways such as "lipid transport", "lipoprotein metabolic process", "Alanine, aspartate and glutamate metabolism", and "Arginine biosynthesis" were significantly enriched in this study. The reliability of the RNA-Seq results was verified by qRT-PCR. These findings provide new insights into the molecular mechanisms of meat quality in sheep. • The chemical composition and amino acid content of muscle were investigated. • Measuring the morphological characteristics of the longissimus thoracis muscle fibers. • RNA-Seq and Label-free were used to explore meat quality related mechanisms in sheep. • qRT-PCR was performed to verify the RNA-Seq results. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the relationship between growth performance, body composition, and fat deposition factors, and feed efficiency in growing lambs. We measured average daily feed intake (ADFI) and body weight (BW) from 653 Hu sheep that were fed a pellet diet. The residual feed intake (RFI) not significantly genetic and phenotypic correlated with the metabolic body weight (MBW) and average daily gain (ADG), but it was significantly genetic and phenotypic correlated with ADFI and the feed conversion ratio (FCR) (p < 0.01). However, the FCR was significantly associated with growth traits (p < 0.01). With the same ADG, body fat deposition was greater in animals with low feed efficiency compared with high feed efficiency. Therefore, excessive fat deposition can affect the feed efficiency of the body, and organ weight and gut-weight have a greater impact on the feed efficiency of lambs. The reticulum stomach and jejunum of lambs with a low RFI were smaller compared with that in the high RFI, indicating that lambs with a low RFI have less intake and a higher absorption rate. Small organs, such as the liver, of lambs with high FE might be associated with low energy expenditure and slow metabolism. This study provides a new perspective to study the biological processes responsible for feed efficiency variation in lambs. [ABSTRACT FROM AUTHOR]

• Both ELOVL5 and FASN genes was associated with the traits of tail fat deposition. • ELOVL5 and FASN expression levels were highest in tail fat. • ELOVL5 expression levels in the big-tail group were highest. • The expression of FASN in small-tail group were highest. • ELOVL5 and FASN have the highest expression at 6 months and 3 months of age. Reducing tail fat deposition can increase the economic value of a carcass and improve feed efficiency. This study aimed to explore ELOVL5 and FASN polymorphisms associated with tail fat deposition and their expression levels of sheep. Association analysis showed that ELOVL5 synonymous mutation g.62534 C > T was associated with tail width, tail fat weight, and relative tail fat weight (P < 0.05). FASN synonymous mutation g.12694 A > G was associated with tail length and width (P < 0.05). Combined effect analyses indicated significant differences between the combined genotypes and tail fat deposition. Quantitative real-time reverse transcription PCR indicated that the ELOVL5 and FASN expression levels were significantly higher in tail fat than in other tissues (P < 0.05). ELOVL5 expression levels in tail-fat tissue of big-tail sheep was significantly higher than that in small-tail sheep (P < 0.01). FASN expression levels were significantly higher in tail-fat tissue of small-tail sheep than in that of big-tail sheep (P < 0.05). During development, ELOVL5 tail fat expression increased significantly from 0 to 6 months old (P < 0.05), and FASN expression at 3 months old was significantly higher than that at 0 (minimum) and 6 months old (P < 0.05). Therefore, ELOVL5 and FASN polymorphisms could represent new candidate molecular markers and targets to reduce tail fat deposition in sheep. [ABSTRACT FROM AUTHOR]

• The SNP of CTNNA3 gene was significantly associated with growth traits. • The SNP of CAP2 gene was significantly associated with body height. • Combined genotypes of CTNNA3 and CAP2 genes was significantly associated with growth traits. • CTNNA3 expression levels in the liver and lung were highest. • CAP2 expression levels in the bone and liver were highest. Growth traits is a critical economic trait for animal husbandry. In this study, the SNPs of CTNNA3 and CAP2 genes were investigated to check whether they are associated with growth traits (body weight, body height, body length and chest circumference) in Hu sheep. The result of the association analysis indicated that the mutation in CTNNA3 (g.2018018 A > G) were associated significantly with body weight, body height, body length and chest circumference (P < 0.05), the mutation in CAP2 (g.8588 T > C) were associated significantly with body height at 140, 160, 180 days (P < 0.05), AA and CC of CTNNA3 and CAP2 were the dominant genotypes associated with growth traits in Hu sheep. Moreover, combined effect analyses indicated that the growth traits with combined genotypes AA CTNNA3 -CC CAP2 and AA CTNNA3 -CT CAP2 were higher than those with genotype GG CTNNA3 -CT CAP2. RT-qPCR indicated that CTNNA3 expression levels were significantly higher in liver and lung than in other nine tissues (P < 0.05), CAP2 expression levels were significantly higher in bone, heart, liver, lung and duodenum than in other six tissues (P < 0.05). In conclusion, CTNNA3 and CAP2 polymorphisms could be used as genetic markers for improving growth traits in Hu sheep husbandry. [ABSTRACT FROM AUTHOR]

BACKGROUND RESULTS CONCLUSION Consumers' preferences for lamb meat vary greatly depending on the specific cut. Variations in the chemical composition across different muscle types play a crucial role in determining meat quality, particularly with regard to flavor. Therefore, it is essential to study the variations in chemical composition among different muscle types in lamb, as well as the mechanisms behind their formation, aiming to understand the flavor variation across the muscle types.Flank muscles showed significantly higher intramuscular fat content and muscle fiber diameter compared to triceps brachii and biceps femoris (BF), at the same time as displaying a significantly lower percentage of type I muscle fibers. Forty‐three differentially abundant volatile compounds (DAVC) were identified across five muscles, with the majority of DAVCs being more abundant in the BF. In total, 161 differentially abundant lipids were identified across five muscles, with triglycerides (TG), phosphatidylcholines (PC), phosphatidyl ethanolamines (PE) and phosphatidylmethanol (PMeOH) showing a strong correlation with DAVCs. A lipid‐gene regulatory network was established, encompassing 664 lipids and 11 107 genes, leading to the identification of pathways and genes that regulate the metabolism of PEs, PMeOH, PCs and TGs.The present study showed the significant variation in flavor compounds among the five edible muscles, as well as the potential reasons for their formation. The results potentially provide a theoretical foundation for improving the meat quality of lamb. © 2024 Society of Chemical Industry. Published by John Wiley & Sons Ltd [ABSTRACT FROM AUTHOR]

Lipoprotein lipase (LPL) is regarded as a candidate gene to regulate fatty acid (FA) composition. The study examined the mRNA expression levels of LPL in different tissues, and conducted a genetic association between the FA composition of the longissimus dorsi muscle of 1029 Hu sheep and the single nucleotide polymorphisms (SNPs) in the LPL gene, as well as the combined genotypes and haplotypes. The results showed that LPL had the highest expression levels in tail fat and subcutaneous fat. Eight SNPs were identified and successfully typed in the intronic region of the LPL gene. Correlation analysis showed that all eight SNPs were related to FA composition. Among them, the CC genotype of SNP1 (g.13879 T>C), the AA genotype of SNP2 (g.13971 A>G), and the TT genotype of SNP3 (g.13993 T>A) had the lowest levels of C8:0 and the highest levels of C18:3n3 content. The TT genotype of SNP6 (g.23413 T>C) and SNP7 (g.23469 T>C) and the AA genotype of SNP8 (g.23472 T>A) had the highest levels of C20:1 content. Linkage disequilibrium analysis showed that SNP1, SNP2, and SNP3 were located in block1, SNP4 (g.17118 A>G) and SNP5 (g.17188 A>G) were located in block2, and SNP6, SNP7, and SNP8 were located in block3. The C18:2n6c content of the TTAAAA combined genotypes in Block1 and the GGGG combined genotypes in Block2 was significantly higher than other combined genotypes. Haplotypes in both Block1 and Block2 significantly affected C18:0, C20:1, and C18:3n3. In conclusion, these genetic markers provide an important source of variation for the selection of FA composition of sheep muscle, which can accelerate the sheep breeding of high-quality meat with a balanced distribution of FA composition. • LPL is expressed at the highest level in adipose tissue. • The SNPs in LPL significantly affect fatty acids. • LPL is involved in the regulation of fatty acid composition. [ABSTRACT FROM AUTHOR]

Background: Circular RNA (circRNA), as a new members of noncoding RNA family, have vital functions in many biological processes by as microRNA sponges or competing endogenous RNAs (ceRNAs). However, little has been reported about the genetic mechanism of circRNAs regulation of feed efficiency in sheep. Results: This study aimed to explore the expression of circRNAs in the liver of Hu sheep with High-RFI (High residual feed intake) and Low-RFI (Low residual feed intake) using transcriptome sequencing. A total of 20,729 circRNAs were identified in two groups, in which 219 circRNAs were found as significantly differentially expressed. Several circRNAs were validated by using RT-PCR, sanger sequencing and RT-qPCR methods. These results demonstrated that the RNA-seq result and expression level of circRNAs identified are reliable. Subsequently, GO and KEGG enrichment analysis of the parental genes of the differentially expressed (DE) circRNAs were mainly involved in immunity response and metabolic process. Finally, the ceRNA regulatory networks analysis showed that the target binding sites for miRNA such as novel_41, novel_115, novel_171 and oar-miR-485-3p in the identified DE cirRNAs. Importantly, two metabolic (SHISA3 and PLEKHH2) and four (RTP4, CD274, OAS1, and RFC3) immune-related target mRNAs were identified from 4 miRNAs. Association analysis showed that the polymorphism (RTP4 c.399 A > G) in the target gene RTP4 were significantly associated with RFI (P < 0.05). Conclusions: Analysis of sequencing data showed some candidate ceRNAs that may play key roles in the feed efficiency in sheep by regulating animal immune and metabolic. These results provide the basis data for further study of the biological functions of circRNAs in regulating sheep feed efficiency. [ABSTRACT FROM AUTHOR]

The aims of this study were to analyze the effects of PPARGC1B and ZEB2 polymorphisms on the body weight of Hu sheep. DNA sequencing and KASPar technologies were used to detect single nucleotide polymorphisms (SNPs) within the PPARGC1B and ZEB2 genes of Hu sheep (n = 207). Two synonymous mutations, g.300 G > A and g.645 C > T, were detected in PPARGC1B and ZEB2, respectively. The body weights of sheep were recorded at 80, 100, 120, 140, 160 and 180-days, and associations between these polymorphisms and body weight changes were analyzed. Association analysis demonstrated that the polymorphisms in PPARGC1B and ZEB2 significantly associated with body weight (p < 0.05). At the g.300 G > A locus, individuals with the GG genotype had significantly higher body weight than those with the AA genotype, and at the g.645 C > T locus, individuals with the TT genotype had significantly higher body weight than those with the TC genotype. Individuals with both polymorphisms exhibited significantly different growth (p < 0.05). These data suggest that polymorphisms in the PPARGC1B and ZEB2 genes can be used as candidate molecular markers for the breeding of desirable growth traits in Hu sheep. [ABSTRACT FROM AUTHOR]

Herein we detected single nucleotide polymorphisms in MEF2B and UCP3 by DNA sequencing and the KASPar technology and analyzed their association with sheep growth traits. Two synonymous mutations, g.1826 C > T and g.10266 G > C, were detected, respectively, and they were found to be significantly associated with sheep growth traits (p < 0.05). In case of MEF2B g.1826 C > T, the average body weight and chest and cannon circumference of sheep with the CC genotype were significantly higher than those of sheep with the CT and TT genotypes (p < 0.05). Moreover, in case of UCP3 g.10266 G > C, the average body weight and chest and cannon circumference of sheep with the GG genotype were significantly higher than those of sheep with the GC and CC genotypes (p < 0.05). Moreover, the average body weight of sheep with the CC/GG genotype was higher compared with those of other genotype combinations. We also assessed MEF2B and UCP3 expression in different sheep tissues, confirming their expression in all examined tissues. To summarize, we believe that the polymorphisms identified in MEF2B and UCP3 can serve as molecular markers for sheep growth traits. [ABSTRACT FROM AUTHOR]

Improving feed efficiency would increase profitability for producers. The objective of this study was to detect the expression levels of ME1 and CA1 and the polymorphisms of ME1 and CA1 associated with the feed conversion ratio (FCR) and residual feed intake (RFI) of Hu sheep by using qRT‐PCR, pooled DNA sequencing and KASPar assay. The qRT‐PCR results indicated that the expression levels of ME1 and CA1 were significantly higher in the liver tissues of low‐RFI sheep than in those of the high‐RFI sheep (p <.01). Association analysis demonstrated that the polymorphism ME1 g.453 C > T was significantly associated with FCR and RFI (p <.05). The polymorphism CA1 g.199 C > T had a significantly associated with FCR (p <.05) and no association with RFI (p >.05). Significant differences were observed between the combined genotypes and FCR and RFI at different measurement periods (p <.05). Thus, we propose the use of these two polymorphisms as new candidate molecular markers for improving feed efficiency in sheep populations. [ABSTRACT FROM AUTHOR]

The number of ribs is an important economic trait in the sheep industry when the sheep are raised for mutton. However, in sheep, the genetic mechanisms regulating rib number are poorly understood. In the present study, we aimed to identify important candidate genes that affect the increase in rib number in sheep. Whole-genome resequencing of 36 Hu sheep with an increased number of ribs (R14) and 36 sheep with normal (R13) rib numbers was carried out. Analysis using three methods (fixation index (F ST), Fisher's exact test, and Chi-squared test) showed that 219 single nucleotide polymorphism sites overlapped among the results of the three methods, which represented 206 genes. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated that the genes were mainly associated with regulation of developmental process, inorganic anion transport, cellular biosynthetic process, tight junction, the oxytocin signaling pathway, and arrhythmogenic right ventricular cardiomyopathy. Four mutations were selected according to the significantly selected genomic regions and important pathways for genotyping and association analysis. The result demonstrated that three synonymous mutations correlated significantly with the rib number. Importantly, we revealed that the CPOX (encoding coproporphyrinogen oxidase), KCNH1 (encoding potassium voltage-gated channel subfamily H member 1), and CPQ (encoding carboxypeptidase Q) genes have a combined effect on rib number in Hu sheep. Our results identified candidate molecular markers for rib number in sheep breeding. • The major candidate genes associated with the rib number were identified using whole-genome resequencing in Hu sheep. • A total of 219 single nucleotide polymorphism sites were found using three methods. • Four single nucleotide polymorphism sites were selected for subsequent genetic effect analysis in a large population. [ABSTRACT FROM AUTHOR]

Testicular size is an excellent proxy for selecting high-fertility rams. The hypothalamus-pituitary-gonadal (HPG) axis plays an important role in regulating reproductive capacity in vertebrates, while key genes and regulatory pathways within the HPG axis associated with testicular size remain largely unknown in sheep. This study comprehensively compared the transcriptomic profiles in the hypothalamus, pituitary and testis of rams after sexual maturity between the large-testis group (LTG, testicular weight = 454.29 ± 54.24 g) and the small-testis group (STG, testicular weight = 77.29 ± 10.76 g). In total, 914, 795 and 10518 differentially expressed genes (DEGs) were identified in the hypothalamus, pituitary and testis between LTG and STG, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that these DEGs were mainly involved in the biological processes of reproduction, biological regulation, and development process. Notably, the neuroactive ligand-receptor interaction and cAMP signaling pathways, commonly enriched by the DEGs in the hypothalamus and pituitary between two groups, were considered as two key signal pathways regulating testicular development through the HPGs axis. Weighted gene co-expression network analysis (WGCNA) identified two modules that were significantly associated with testicular size, and 97 key genes were selected with high module membership (MM) and gene significance (GS) in these two modules. Finally, a protein-protein interaction (PPI) network was constructed, and ten genes with the highest degree were represented as hub genes, including FOS , NPY , SST , F2 , AGT , NTS , OXT , EDN1 , VIP and TAC1. Taken together, these results provide new insights into the molecular mechanism underlying the HPG axis regulating testicular size of Hu sheep. • A total of 914, 795 and 10518 DEGs were identified in the HPG axis-related tissues between LTG and STG, respectively. • The DEGs were mainly involved in the biological processes of reproduction, biological regulation and development process. • Ten hub proteins were identified from a PPI network, including FOS , NPY , SST , F2 , AGT , NTS , OXT , EDN1 , VIP and TAC1. [ABSTRACT FROM AUTHOR]

The testis is an important organ for maintaining fertility in males, and testis size is positively correlated with ejaculate volume, sperm motility, thus fertility. Spermatogenesis-associated 6 (SPATA6) is an evolutionarily conserved testis-specific gene reported in many species. However, the effect of SPATA6 expression levels on testicular development and the effect of single nucleotide polymorphisms (SNPs) on testis and epididymis phenotype in sheep have not been studied. The purpose of the research was to investigate the expression profile of SPATA6 and its effect on testicular development and to confirm the effect of SNPs on the testis and epididymis phenotype. In this study, we detected a 1245bp coding sequence (CDS) of SPATA6 and encoded 414 amino acids. The expression levels of SPATA6 were significantly higher in the testis than in other tissues and gradually increased with testis development. Moreover, the expression level in the large testis was significantly higher than that in the small testis at six months. A total of 11 SNPs were detected in the coding region of SPATA6 by cDNA-pooling sequencing and improved multiplex ligation detection reaction (iMLDR) methods. Correlation analysis showed that SNP2 (c. 3631C > G) significantly affected left epididymis weight (LEW) and right epididymis weight (REW), and SNP10 (c. 937 A > G) significantly affected REW. And the combined genotype of SNP1 (c. 4245 G > A) and SNP2 significantly affected REW. The current study concluded that SPATA6 plays an important role in testicular development and the SNPs significantly associated with the epididymis phenotype can provide molecular markers for the early selection of high-fertility Hu sheep. • SPATA6 plays an important role in testicular development. • 11 SNPs were identified in the SPATA6 gene mRNA sequence. • Two SNPs were associated with epididymal phenotype in Hu sheep. [ABSTRACT FROM AUTHOR]

Sheep milk is the most important feed resource for newborn lambs and an important food resource for humans. Sheep milk production and ingredients are influenced by genetic and environmental factors. In this study, we implemented selection signature analysis using Illumina Ovine SNP50 BeadChip data of 78 meat Lacaune and 103 milk Lacaune sheep, which have similar genetic backgrounds, from the Sheep HapMap project to identify candidate genes related to ovine milk traits. Since different methods can detect different variation types and complement each other, we used a haplotype-based method (hapFLK) to implement selection signature analysis. The results revealed six selection signature regions showing signs of being selected (P<0.001): chromosomes 1, 2, 3, 6, 13 and 18. In addition, 38 quantitative trait loci (QTLs) related to sheep milk performance were identified in selection signature regions, which contain 334 candidate genes. Of those, SUCNR1 (succinate receptor 1) and PPARGC1A (PPARG coactivator 1 alpha) may be the most significant genes that affect sheep milking performance, which supply a significant indication for future studies to investigate candidate genes that play an important role in milk production and quality. [ABSTRACT FROM AUTHOR]

The content of intramuscular fat (IMF) is one of the most important factors that has a large impact on meat quality, and it is an effective way to improve IMF according to marker-assisted selection (MAS). Fatty-acid synthase (FASN) is a key gene in meat lipid deposition and fatty acid composition. Thus, this study was conducted to investigate the expression profile of FASN in mRNA and protein levels using real-time quantitative PCR (RT-qPCR) and western-blot methods. In addition, single nucleotide polymorphisms (SNPs) within FASN in 921 Hu rams with IMF content records were investigated using DNA-pooling sequencing and improved multiple ligase detection reaction (iMLDR) methods. Consequently, the highest mRNA expression level of FASN was observed in the perinephric fat, and the lowest in the liver among the 11 tissues analyzed, while no significant difference was found in mRNA and protein expression levels in longissimus dorsi among individuals with different IMF contents. A total of 10 putative SNPs were identified within FASN, and 9 of them can be genotyped by iMLDR method. Notably, two SNPs were significantly associated with IMF content, including NC_040262.1: g.5157 A > G in intron 5 (p = 0.046) and NC_040262.1: g.9413 T > C in intron 16 (p = 0.041), which supply molecular markers for improving meat quality in sheep breeding. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the differences in the anti-oxidant capabilities and related gene expressions of six-month-old Hu sheep with different testis sizes. A total of 201 Hu ram lambs were fed up to 6 months in the same environment. Based on their testis weight and sperm count, 18 individuals were selected and divided into large (n = 9) and small (n = 9) groups, with an average testis weight of 158.67 g ± 5.21 g and 44.58 g ± 4.14 g, respectively. The total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD), and malondialdehyde (MDA) concentration in testis tissue were tested. The localization of antioxidant-related genes, GPX3 and Cu/ZnSOD in testis were detected by immunohistochemistry. The GPX3, Cu/ZnSOD expression, and relative mitochondrial DNA (mtDNA) copy number were detected by quantitative real-time PCR. Compared with the small group, the T-AOC (2.69 ± 0.47 vs. 1.16 ± 0.22 U/mgprot) and T-SOD (22.35 ± 2.59 vs. 9.92 ± 1.62 U/mgprot) in the large group were significantly higher, whereas the MDA (0.72 ± 0.13 vs. 1.34 ± 0.17 nM/mgprot) and relative mtDNA copy number in the large group was significantly lower (p <.05). Immunohistochemistry results indicated that the GPX3 and Cu/ZnSOD were expressed in Leydig cells and seminiferous tubule. The expressions of GPX3 and Cu/ZnSOD mRNA in the large group were significantly higher than those in the small group (p <.05). In conclusion, Cu/ZnSOD and GPX3 widely expressed in the Leydig cells and seminiferous tubule, high expression of Cu/ZnSOD and GPX3 in a large group has a higher potential in addressing oxidative stress and contribute to spermatogenesis. [ABSTRACT FROM AUTHOR]

The early weaning and starter feeding have significant effects on lamb growth and digestive tract development. However, it is not clear whether the expression of IGF-1/IGF-1R genes could be affected by feeding starter and weaning. Therefore, a total of 102 Hu male neonatal lambs were randomly divided into 3 groups: fed with starter at 42d + weaned at 56d (group A), fed with starter at 7d + weaned at 28d (group B) and fed with starter at 7d + weaned at 56d (group C), to explore the effects of starter feeding and weaning age on developmental expressions of IGF-I gene in liver and IGF-IR gene in rumen of Hu sheep. The results showed that IGF-I and IGF-IR genes were expressed extensively in various tissues of lambs, the expression of IGF-I was significantly higher in liver (p < 0.01), while the expression of IGF-IR was higher in rumen among gastrointestinal tissues. The mRNA level of IGF-I of group C was strikingly higher than that of group A at 28, 70, and 84d (p < 0.01) in liver, respectively. Rumen mRNA level of IGF-IR of group C was prominently higher than that of group A at 70d and 84d (p < 0.05), respectively. However, IGF-IR expression of group A was higher than that of group C at 14 and 42d (p < 0.05). The mRNA level of IGF-I of group B was significantly higher than that of group C at 42, 56, 70, and 84d (p < 0.01) in liver, while IGF-IR expression of group B was higher than that of group C at 70d (p < 0.01). In conclusion, early weaning and starter feeding affected liver IGF-I mRNA and rumen IGF-IR mRNA expression. [ABSTRACT FROM AUTHOR]

Protein tyrosine phosphatase non-receptor type 3 (PTPN3), a member of the membrane-associated non-receptor protein tyrosine phosphatase (PTP) family, plays significant roles in the cytoplasm and affects the development and growth of skin and hair. A recent study identified the PTPN3 as the potential gene related to sheep wool quality. To detect single-nucleotide polymorphisms (SNPs) of PTPN3 and elucidate its association with wool production and growth traits in fine wool sheep a total of 644 healthy SG (South African mutton merino♂ × Gansu alpine fine-wool sheep♀, SG) and SSG (South African mutton merino♂ × SG♀, SSG) hybrid sheep were selected. Pooled-DNA sequencing and SNPscan methods were used to scan and genotype SNPs within PTPN3. Association analyses between SNPs and wool production and growth traits were implemented. Consequently, the results revealed that PTPN3 has six SNPs (two missense mutations, one synonymous mutation, and three intron mutations), of which four loci (SNP2, SNP3, SNP4, and SNP5) were significantly positively correlated with growth and wool traits (p < 0.05). SNP4 was significantly (p < 0.05) linked with thigh wool length, and SNP6 was significantly (p < 0.05) associated with abdomen wool length. Moreover, one strongly linked SNP block was identified to be correlated with wool production and growth traits (body weight and body size). The significant SNPs founded by this study could serve as useful genetic markers for breeding fine-wool sheep.® [ABSTRACT FROM AUTHOR]

Understanding the effects of weaning on the changes of digestive function could help to design efficient intervention strategies for promoting the development of the lamb during the early stages of life. In this study, 24 lambs were divided into two groups (control group, lambs were not weaned; and weaning group, lambs were weaned at 21 days of age). The growth, nutrient digestion, gastrointestinal enzyme activity, plasma biochemical indicators, and intestinal microbiota at 7–49 days were determined, as well as the impact of early weaning. The nutrient digestion changed rapidly with age, especially at 14–28 days (p < 0.05). Weaning reduced the dry matter (DM), crude protein (CP), and ether extract (EE) intake and digestion, but increased the starch, neutral detergent fiber (NDF), and acid detergent fiber (ADF) intake and digestion (p < 0.05). Weaning did not affect the overall jejunal microbiota (p > 0.05), but affected the relative abundance of certain bacteria taxa (p < 0.05). Lactic acid-producing bacteria, such as Olsenella, Bacillus, Sharpea, and Bifidobacterium are closely related to CP or EE digestion and growth performance (p < 0.05). In summary, we delineated the pattern of nutrient digestion and intestinal microbiota development in young lambs, and the impact of early weaning. [ABSTRACT FROM AUTHOR]

Small ruminants farming plays an important role in the livelihood of a large part of the population. Herein we aimed to analyze the effects of single nucleotide polymorphisms in PRKAA2 gene on the growth-related traits of Hu sheep and Dorper sheep. The body weight and body type of 1254 sheep were measured at 80, 100, 120, 140, 160 and 180d, and 37620 phenotypic data were collected. RT-qPCR analysis was performed to test PRKAA2 gene expressed in different tissues of sheep, with the highest expression level in spleen, followed by kidney. In the present study, the PRKAA2 gene sequencing revealed one polymorphism located on Chr1 (Oar_rambouillet_v1.0), termed as chr1:32832382 G > A, and were significantly associated with growth traits of sheep (p < 0.05). The body weight, body length, chest circumference, and cannon circumference of individuals with AA genotype were significantly higher than those with the GG and GA genotypes (p < 0.05). Our findings reveal that PRKAA2 gene could be used as a marker-assisted selection to improve the growth-related traits of sheep. [ABSTRACT FROM AUTHOR]

This study aimed to analyze the effects of polymorphisms in GP5 on blood physiological indices of 1065 sheep. The coefficients of variation of the red blood cell count (RBC), hemoglobin concentration (HGB), mean platelet volume (MPV), mean erythrocyte hemoglobin content (MCH), and red blood cell distribution-coefficient of variation (RDW-CV) were greater than 10%, and there was a very significant correlation between the main indices such as RBC, white blood cell, and platelet count (PLT) and most other indices (p < 0.01). qRT-PCR showed that GP5 was expressed in the heart, liver, spleen, lung, kidney, rumen, duodenum, muscle, tail fat, and lymph tissue, with significantly higher expression in the lymph. Subsequently, we detected single nucleotide polymorphisms (SNPs) in GP5 from group, which identified synonymous mutation g.657 T > C in the first exon of GP5. Association analysis showed significant correlations between the SNP and the physiological traits (p < 0.05), in which the RBC, neutrophilic granulocyte (NEUT) and RDW-CV values in sheep with the TC genotype and TT genotype were markedly lower than those in the CC genotype (p < 0.05). Thus, GP5 polymorphisms could be candidate biomarkers to screen blood physiological indices. [ABSTRACT FROM AUTHOR]

Hu sheep is an excellent short fat-tailed breed in China. Fat deposition in Hu sheep tail affects carcass quality and consumes a lot of energy, leading to an increase in feed cost. The objective of this study was to analyze the effects of HMGA1 polymorphism on tail fat weight in Hu sheep. Partial coding and non-coding sequences of HMGA1 were amplified with PCR and single nucleotide polymorphisms (SNP) of HMGA1 in 1163 Hu sheep were detected using DNA sequencing and KASPar technology. RT-qPCR analysis was performed to test HMGA1 expression in different tissues. The results showed that the expression of HMGA1 was higher in the duodenum, liver, spleen, kidney, and lung than in the heart, muscle, rumen, tail fat, and lymph. A mutation, g.5312 C > T, was detected in HMGA1; g.5312 C > T was significantly associated with tail fat weight, relative weight of tail fat (body weight), and relative weight of tail fat (carcass) (p < 0.05). The tail fat weight of the TT genotype was remarkably higher than that of the CC and TC genotypes. Therefore, HMGA1 can be used as a genetic marker for marker-assisted selection of tail fat weight in Hu sheep. [ABSTRACT FROM AUTHOR]

Omega-3 polyunsaturated fatty acids (ω-3 PUFAs), such as α-linolenic acid (ALA), eicosapentaenoic acid, and docosahexaenoic acid, are involved in male reproductive function. In this study, we investigated the effects of linseed oil (LO) as a source of ALA on the steroidogenesis and changes of testicular histology in rams. Sixteen 3-month old rams during peri-puberty were randomly assigned into two groups. Eight rams were assigned as the control group, and the other received LO (4% dry matter of total diet) as the LO treatment group. After an 81-day feeding trial, the rams were slaughtered and investigated. Results revealed that compared with control group, diet containing LO did not affect body weight (36.87 ± 0.53 kg vs. 37.65 ± 0.64 kg, respectively; P = 0.361), average daily gain (227.47 ± 5.82 g vs. 237.95 ± 9.22 g, respectively; P = 0.353) and epididymis weight (40.77 ± 4.41 g vs. 45.53 ± 4.01 g, respectively; P = 0.398), however, it up-regulated PUFAs metabolism and steroidogenesis-related genes mRNA expression (P < 0.05), and increased plasma estradiol concentration (14.88 ± 0.67 pg/mL vs. 19.50 ± 1.27 pg/mL, respectively; P < 0.05). Therefore, LO stimulated seminiferous tubule development and increased the number of Sertoli cells (19.17 ± 2.14 vs. 27.2 ± 2.39, respectively; P < 0.01), germ-cell layers, as well as testis weight (148.65 ± 22.66 g vs. 249.96 ± 30.63 g, respectively; P < 0.05). All these results suggested that LO can improve testis development during peri-puberty by regulating steroidogenesis in rams' testes. [ABSTRACT FROM AUTHOR]

CD274 and PLEKHH2 genes have been identified as immune- and multiple diseases-related genes, and have recently garnered significant interest. However, their role in regulating immune functions in sheep remains largely unexplored. In this study, we aimed to investigate the effects of polymorphisms in CD274 and PLEKHH2 on hematologic parameters in 915 sheep. Our results showed that the CD274 and PLEKHH2 genes were most highly expressed in the spleen and tail fat, respectively, as determined by qRT-PCR. We also identified a G to A mutation (g 0.11858 G > A) in the exon 4 region of CD274, and a C to G mutation (g 0.38384 C > G) in the intron 8 region of PLEKH2. Association analysis revealed that CD274 g 0.11858 G > A was significantly associated with RBC, HCT, MCHC, and MCV (P < 0.05), while PLEKHH2 g 0.38384 C > G was significantly associated with HCT, MPV, MCHC, and MCV (P < 0.05). These results suggest that CD274 and PLEKHH2 genes may play a role in regulating blood physiological indicators and could be potential functional candidates for influencing immune traits in sheep breeding programs. • The SNPs of CD274 and PLEKHH2 genes were associated with hematologic parameters in sheep. • Combined genotypes of CD274 and PLEKHH2 genes were associated with hematologic parameters in sheep. • The ovine CD274 and PLEKHH2 genes had the highest mRNA expression in the spleen and tail fat, respectively. [ABSTRACT FROM AUTHOR]

ADP-dependent glucokinase (ADPGK) plays an important role instead of hexokinase in regulating energy metabolism via the Embden–Meyerhof–Parnas Pathway. And energy provided via glycolysis promotes testis development and spermatogenesis. In this study, 466 Hu sheep were screened for mutations in the ADPGK gene to examine the association of the ADPGK gene polymorphisms with the testis traits and spermatogenesis. The NC_056060.1: g.31295 C > T SNP was found in the 3′-UTR region, resulting in two genotypes CC and TC type with genotypic frequencies of 0.66 and 0.34, respectively. This mutation was significantly associated with testis weight, testis long circumference, testis short girth, epididymis weight, and sperm concentration (p < 0.05). Moreover, TC genotype individuals had an increased tendency in the expression of the ADPGK gene and had significant reproductive performance advantages compared with CC genotype individuals in the study. And compared with the small testes (<50 g), the ADPGK gene expression of big testes (>160 g) increased significantly. This indicates an association between the ADPGK gene and reproductive organ parameters and sperm count in selected Hu sheep breed, and this SNP may serve as an effective DNA molecular marker for marker-assisted selection in Hu sheep breeding programs. [ABSTRACT FROM AUTHOR]

The development of the rumen is a critical physiological challenge in newborn ruminants. However, the molecular mechanism underlying different stages of rumen development in sheep remains poorly understood. Here, RNA sequencing and bioinformatics analysis were performed to compare the transcription profiles of rumen development at 7, 28 and 56 days of birth (D7, D28 and D56). We identified 1246, 2257 and 627 differentially expressed genes (DEGs) between D7 and D28, between D7 and D56, between D28 and D56, respectively. Also, 70 DGEs were co-expressed at these three time points. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated most DEGs mainly related to transporter activity, channel activity and metabolism pathways. Noteworthy, the expression levels of most genes (CA4, CA9, CA12 and CA14) in nitrogen metabolic pathways were negatively correlated with the papilla length and width, but the papilla length and width were positively correlated with the expression of genes (PLA2G3, SLC26A9, SLC34A3) in ion transport pathway, suggesting that these genes may be involved in nitrogen metabolic and ion transport pathway and thus affect rumen development. These results provide new insight into the changes in RNA expression at different time points of rumen development in Hu sheep. [ABSTRACT FROM AUTHOR]

Corn straw and corncobs contain large amounts of crude fibers and are widely used in mutton sheep husbandry in northwest China. The aim of this study was to determine whether feeding with corn straw or corncobs affects lamb testis development. A total of 50 healthy Hu lamb at two-month-old (average body weight of 22.3 ± 0.1 kg) were randomly and equally divided into two groups, and the lambs were equally allocated to five pens in each group. The corn straw group (CS) received a diet containing 20% corn straw, whereas the corncobs group (CC) received a diet containing 20% corncobs. After a 77-day feeding trial, the lambs, except the heaviest and lightest in each pen, were humanely slaughtered and investigated. Results revealed no differences in body weight (40.38 ± 0.45 kg vs. 39.08 ± 0.52 kg) between the CS and CC groups. Feeding diet containing corn straw significantly (P < 0.05) increased testis weight (243.24 ± 18.78 g vs. 167.00 ± 15.20 g), testis index (0.60 ± 0.05 vs. 0.43 ± 0.04), testis volume (247.08 ± 19.99 mL vs. 162.31 ± 14.15 mL), diameter of seminiferous tubule (213.90 ± 4.91 μm vs. 173.11 ± 5.93 μm), and the number of sperm in the epididymis (49.91 ± 13.53 × 108/g vs. 19.34 ± 6.79 × 108/g) compared with those in the CC group. The RNA sequencing results showed 286 differentially expressed genes, and 116 upregulated and 170 downregulated genes were found in the CS group compared with the CC group. The genes affecting immune functions and fertility were screened out. Corn straw decreased the mtDNA relative copy number in the testis (P < 0.05). These results suggest that compared with corncobs, feeding corn straw in the early reproductive development stage of lambs increased the testis weight, diameter of seminiferous tubule and the number of cauda sperm. [Display omitted] • There were 286 DEGs in testis between corn straw and corncobs groups. • The genes closely related to testis function were screened out. • Feeding corn straw in the early reproductive development stage of lambs increased the number of sperm in the epididymis. [ABSTRACT FROM AUTHOR]

Background: Ferulic acid esterase (FAE)-secreting Lactiplantibacillus plantarum A1 (Lp A1) is a promising silage inoculant due to the FAE's ability to alter the plant cell wall structure during ensiling, an action that is expected to improve forage digestibility. However, little is known regarding the impacts of Lp A1 on rumen microbiota. Our research assessed the influences of Lp A1 in comparison to a widely adopted commercial inoculant Lp MTD/1 on alfalfa's ensilage, in vitro rumen incubation and microbiota. Results: Samples of fresh and ensiled alfalfa treated with (either Lp A1 or Lp MTD/1) or without additives (as control; CON) and ensiled for 30, 60 and 90 d were used for fermentation quality, in vitro digestibility and batch culture study. Inoculants treated silage had lower (P < 0.001) pH, acetic acid concentration and dry matter (DM) loss, but higher (P = 0.001) lactic acid concentration than the CON during ensiling. Compared to the CON and Lp MTD/1, silage treated with Lp A1 had lower (P < 0.001) aNDF, ADF, ADL, hemicellulose, and cellulose contents and higher (P < 0.001) free ferulic acid concentration. Compared silage treated with Lp MTD/1, silage treated with Lp A1 had significantly (P < 0.01) improved ruminal gas production and digestibility, which were equivalent to those of fresh alfalfa. Real-time PCR analysis indicated that Lp A1 inoculation improved the relative abundances of rumen's total bacteria, fungi, Ruminococcus albus and Ruminococcus flavefaciens, while the relative abundance of methanogens was reduced by Lp MTD/1 compared with CON. Principal component analysis of rumen bacterial 16S rRNA gene amplicons showed a clear distinction between CON and inoculated treatments without noticeable distinction between Lp A1 and Lp MTD/1 treatments. Comparison analysis revealed differences in the relative abundance of some bacteria in different taxa between Lp A1 and Lp MTD/1 treatments. Silage treated with Lp A1 exhibited improved rumen fermentation characteristics due to the inoculant effects on the rumen microbial populations and bacterial community. Conclusions: Our findings suggest that silage inoculation of the FAE-producing Lp A1 could be effective in improving silage quality and digestibility, and modulating the rumen fermentation to improve feed utilization. [ABSTRACT FROM AUTHOR]

The screening of genomic variations within the male-specific region of the mammalian Y chromosome (MSY) is one of the most effective ways to investigate paternal evolutionary history and identify molecular markers related to male fertility. The current study was to identify single nucleotide polymorphisms (SNPs) within single-copy genes of the ovine MSY, and confirm whether they are associated with testicular size. A total of 21 Y-specific gene fragments were successfully amplified to screen Y-SNPs in 956 rams across nine sheep breeds. Three Y-SNPs, including SRY16: g.88 A > G in South African Mutton Merino sheep, ZFY16: g.146 C > T in Suffolk and South African Mutton Merino sheep, and EIF2S3Y2: g.77 C > G in Hu and Tan sheep, were identified using DNA-pooled sequencing and PCR restriction fragment length polymorphism (PCR-RFLP) methods. The investigation of the global distribution for three Y-SNPs showed that the C allele of ZFY16: g.146 C > T co-segregated with haplogroup y-HC, and the C/G allele of EIF2S3Y2: g.77 C > G co-segregated with haplogroup y-HA/y-HB1 in Hu sheep according to data mining from a previous study. In addition, association analysis revealed that ZFY16: g.146 C > T had a significant effect on yearling scrotal circumference in Suffolk sheep, and EIF2S3Y2: g.77 C > G was significantly associated with testicular and epididymis weight in Hu sheep (P ≤ 0.05). The current study concluded that Y-SNPs were associated with testicular size in specific sheep, which provides valuable candidate makers for selecting elite rams at an early age. • 21 Y-specific gene fragments were successfully amplified. • Three SNPs within the ovine MSY were identified by DNA-pooled sequencing and PCR-RFLP methods. • Tow Y-SNPs were associated with testicular size in specific sheep breeds. [ABSTRACT FROM AUTHOR]

Fatty acid (FA) composition contributes greatly to the quality and nutritional value of lamb meat. In the present study, FA was measured in longissimus thoracis (LT) muscles of 1,085 Hu sheep using gas chromatography. Comparative transcriptomic analysis was conducted in LT muscles to identify differentially expressed genes (DEGs) between six individuals with high polyunsaturated fatty acids (H-PUFA, 15.27% ± 0.42%) and six with low PUFA (L-PUFA, 5.22% ± 0.25%). Subsequently, the single nucleotide polymorphisms (SNPs) in a candidate gene PLIN2 were correlated with FA traits. The results showed a total of 29 FA compositions and 8 FA groups were identified, with the highest content of monounsaturated fatty acids (MUFA, 46.54%, mainly C18:1n9c), followed by saturated fatty acids (SFA, 44.32%, mainly C16:0), and PUFA (8.72%, mainly C18:2n6c), and significant correlations were observed among the most of FA traits. Transcriptomic analyses identified 110 upregulated and 302 downregulated DEGs between H-PUFA and L-PUFA groups. The functional enrichment analysis revealed three significant Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and 17 gene ontology (GO) terms, in which regulation of lipolysis in adipocytes, the AMPK signaling pathway, and the PPAR signaling pathway may play important roles in FA metabolism and biosynthesis. In addition, weighted gene co-expression network analysis (WGCNA) identified 37 module genes associated with PUFA-related traits. In general, PLIN1, LIPE, FABP4, LEP, ACACA, ADIPOQ, SCD, PCK2, FASN, PLIN2, LPL, FABP3, THRSP , and ACADVL may have a great impact on PUFA metabolism and lipid deposition. Four SNPs within PLIN2 were significantly associated with FA. Of those, SNP1 (g.287 G>A) was significantly associated with C18:1n9c and MUFA, and SNP4 (g.7807 T>C) was significantly correlated with PUFA (C18:3n3). In addition, the combined genotype of SNP1 (g.287 G>A), SNP3 (g.7664 T>C), and SNP4 (g.7807 T>C) were significantly correlated with C16:1, C17:0, C18:1C6, PUFA (C18:3n3, C22:6n3), and n-6/n-3 PUFA. These results contribute to the knowledge of the biological mechanisms and genetic markers involved in the composition of FA in Hu sheep. [ABSTRACT FROM AUTHOR]

The growth hormone receptor (GHR) is associated with animal growth and development. To investigate such effects on GHR gene expression, a total of 102 Hu lambs were randomly allocated to one of three groups (Group 1: starter diet from 7 d of age, weaning at 56 d of age; Group 2: starter diet from 42 d of age, weaning at 56 d of age; Group 3: starter diet from 7 d of age; weaning at 28 d of age). Six lambs from each group were sacrificed every 14 d to investigate the effects of starter feeding and weaning age on GHR mRNA expression in the liver and rumen. The results revealed that GHR mRNA expression was significantly higher in the liver and rumen (p < 0.05) than in other tissues. Early starter feeding up-regulated hepatic GHR mRNA expression on days 14, 28, 42 and 56 and ruminal GHR mRNA expression on days 28, 42, 70, and 84 (p < 0.05). Early weaning up-regulated hepatic GHR mRNA expression on days 56, 70 and 84 and ruminal GHR mRNA expression on days 42, 56, 70 and 84 (p < 0.05). Dietary and weaning regimes and age affected the hepatic and ruminal GHR mRNA expression. [ABSTRACT FROM PUBLISHER]

Thirty-two male Hu lambs (32.31 ± 3.31 kg; 4-months-old) were randomly assigned to four treatments: (1) control (CON), (2) selenium-enriched yeast (SeY, 0.8 mg/kg), (3) selenized glucose (SeGlu, 0.8 mg/kg), and (4) sodium selenite (SS, 0.8 mg/kg) to evaluate their effects on Hu lamb production and slaughter performance, antioxidant capacity, hematological parameters, meat quality and shelf-life. The production and slaughter performances were not different (P > 0.05) among treatments. SeGlu and SeY increased (P < 0.05) the total antioxidant capacity in serum and muscle selenium content while decreasing (P < 0.05) the malondialdehyde (MDA) contents both in serum and muscle. SeGlu extended muscle shelf-life by 7.7 h compared with CON and decreased (P < 0.05) yellowness (b*) and lightness (L*) in meat stored for 24 h. In summary, the effects of SeGlu were similar to those of SeY and better than those of SS in improving serum and muscle antioxidant status, prolonging muscle shelf-life, and increasing selenium deposition in muscle. • Sodium selenite, Se-yeast, and selenized glucose could raise serum antioxidant capacity and muscle Se content in lambs. • A new organic selenium source, selenized glucose, can be used as a feed additive to produce selenium-enriched meat. • Selenized glucose could improve the antioxidant capacity and prolong the shelf life of lamb meat. [ABSTRACT FROM AUTHOR]

The present study was undertaken to elucidate the direct ALA effects on mice TM4 Sertoli cells proliferation in vitro. Our results showed that TM4 cells viability was significantly stimulated by ALA (p < 0.05). The 50 μM ALA increased the concentration of total antioxidant capacity, induced the mitochondrial membrane hyperpolarized, and markedly decreased the number of apoptosis cells (p < 0.05). ALA also up-regulated G2/Mitotic-specific cyclin-B1 gene and apoptosis suppressive gene Bcl2 expression (p < 0.05). In conclusion, those results indicated that ALA could increase TM4 Sertoli cells antioxidant capacity, induced the mitochondrial membrane hyperpolarized, inhibited cells apoptosis and stimulated TM4 Sertoli cells proliferation in vitro. [ABSTRACT FROM AUTHOR]

Simple Summary: The Chinese Hu sheep is well-known for its high fertility and early sexual maturity. Rams reach sexual maturity at the age of six months. This study investigated differential crude fat content, fatty acid composition and related metabolism genes localization and expression in cauda epididymis between high and low fertility Hu sheep at six months old. The crude fat content was positively correlated with epididymal weight. High expression of SREBP1 and EVOVL2 in high fertility Hu sheep may lead to the higher n-3 PUFA content, especially DHA, providing a basis for understanding the relationship between fertility and sperm storage environment. The epididymis is an organ that transports, matures and stores sperm, and has functions such as secretion and absorption. Polyunsaturated fatty acid (PUFA) compositions in sperm membrane were changed during the process of epididymis maturation and influence the male fertility. This study aimed to investigate differences in crude fat and fatty acid content in cauda epididymis between high and low fertility of Hu sheep. One hundred and seventy-nine Hu ram lambs were fed from 56 days to 6 months under the same environment. After the feeding trial, all rams were slaughtered, and the body weight, testicular weight, epididymal weight and sperm density were measured. Pearson correlation analysis showed significantly moderate positive correlation between epididymal weight and sperm density and testicular weight. Eighteen rams were selected and divided into the high fertility group (H, n = 9) and low fertility group (L, n = 9) according to the epididymal weight, sperm density and histomorphology. The crude fat content, fatty acid profile and genes related to fatty acid metabolism were detected. The crude fat content, total fatty acid, total n-3 PUFA and docosahexaenoic acid (C22:6n-3, DHA) content of cauda epididymis in high fertility group was significantly higher than those in low fertility group (p < 0.05). However, the ratio of n-6/n-3 PUFA was significantly lower than that in group L (p < 0.05). Immunohistochemistry results showed that SREBP1 and ELOVL2 were expressed in pseudostratified columnar ciliated epithelium and smooth muscle cells. The mRNA expression of SREBP1 (p = 0.09) and ELOVL2 (p < 0.05) in the high fertility group were increased. In conclusion, the high expression of SREBP1 and ELOVL2 may contribute to high n-3 PUFA content in cauda epididymis of high-fertility Hu sheep. [ABSTRACT FROM AUTHOR]

Simple Summary: PRAMEY (preferentially expressed antigen in melanoma, Y-linked) is a cancer-testis antigen and is believed to play an important role in testicular development and gametogenesis. However, the structure, expression and function of PRAMEY in sheep remain unknown. In this paper, we found that PRAMEY was extremely expressed in the testis and significantly upregulated during testicular growth. The mRNA abundance of PRAMEY was significantly correlated with testicular size traits in Hu sheep at 6 months. These results imply that PRAMEY may play an important role in testicular growth. PRAMEY (preferentially expressed antigen in melanoma, Y-linked) belongs to the cancer-testis antigens (CTAs) gene family and is predominantly expressed in testis, playing important roles in spermatogenesis and testicular development. This study cloned the full-length cDNA sequence of ovine PRAMEY using the rapid amplification of cDNA ends (RACE) method and analyzed the expression profile and copy number variation (CNV) of PRAMEY using quantitative real-time PCR (qPCR). The results revealed that the PRAMEY cDNA was 2099 bp in length with an open reading frame (ORF) of 1536 bp encoding 511 amino acids. PRAMEY was predominantly expressed in the testis and significantly upregulated during postnatal testicular development. The median copy number (MCN) of PRAMEY was 4, varying from 2 to 25 in 710 rams across eight sheep breeds. There was no significant correlation between the CNV of PRAMEY and testicular size, while a significant positive correlation was observed between the mRNA expression and testicular size in Hu sheep. The current study suggests that the expression levels of PRAMEY were closely associated with testicular size, indicating that PRAMEY may play an important role in testicular growth. [ABSTRACT FROM AUTHOR]

ZNF280BY , a bovid-specific Y chromosome gene, was firstly found to be highly expressed in bovine testis, indicating that it may play important roles in testicular development and male fertility. In this study, we firstly cloned the full-length cDNA of ovine ZNF280BY containing 1,993 bp, and with a 1,632 bp open reading frame. ZNF280BY was predominantly expressed in the testis, and its expression level was significantly higher in large testis than in small testis in Hu sheep at 6 months of age. In addition, the expression level of ZNF280BY significantly increased during testicular development, showing the highest expression level at 12 mo of age. ZNF280BY showed copy number variation (CNV) in 723 rams from eight sheep breeds, ranging from 17 to 514 copies, with a median copy number of 188. Pearson correlation analysis showed that the CNV of ZNF280BY was negatively correlated with testis size in Hu sheep. Furthermore, its mRNA expression level in testis had no significant correlation with the CNV but was significantly correlated with testis size. This study concluded that the expression of ZNF280BY was closely related to testicular development, and the CNV of ZNF280BY could be used as an important genetic marker to evaluate the ram reproductive capacity at an early stage in Hu sheep. [ABSTRACT FROM AUTHOR]

Rumen development is critical for the development of early lambs. This work aims to evaluate the effects of abrupt weaning at day 21 on rumen fermentation, histomorphological traits and the ruminal microbiota compared with continuous suckling. Twelve pairs of artificially reared full-sib neonatal male Hu lambs were allocated to two groups, one of which was weaned at day 21 (EW group) and the other which was not weaned (CON group). At day 26 and day 49, six lambs from each group were randomly selected and sacrificed to collect ruminal contents and rumen tissue samples. Results showed that weaning influenced the fermentation parameters in the rumen, and altered the microbial community composition on day 49 (p < 0.05). Several genera were associated with rumen fermentation parameters (p < 0.05). Volatile fatty acid (VFA) concentration is the key parameter impacting microbiota composition. Weaning influenced the expression of genes associated with VFA metabolism and regulation of cell proliferation (p < 0.05). In conclusion, weaning significantly influenced the morphological and functional development of the rumen, and bacterial community composition. The microbial community composition was strongly associated with rumen weight and fermentation profiles, but not with morphological development. [ABSTRACT FROM AUTHOR]

Simple Summary: Increasing the starch level in the diet of ruminants can promote rumen fermentation to a certain extent and improve production performance. However, the degradation of rapidly fermentable carbohydrates in the rumen can lead to the accumulation of large amounts of organic acids (mainly volatile fatty acids and lactic acid), increasing the risk of subacute ruminal acidosis (SARA) in high-yielding ruminants. By changing the fermentation sites of rapidly fermentable carbohydrates, the rumen environment and feed efficiency could be effectively improved. The object of this study was to explore the effects of replacing corn starch with barley starch in the diet of Hu sheep on the fermentation and microflora in the rumen. Our results showed that, starch in the diet cannot be derived entirely from barley, and it is feasible to replace 33% of the corn with barley in the diet for Hu sheep. This study aimed to explore the effects of different levels of barley starch instead of corn starch on the rumen fermentation and microflora when feeding a corn-based diet to Hu sheep. Thirty-two male Hu sheep equipped with permanent rumen fistulas were selected and fed in individual metabolic cages. All sheep were randomly divided into four groups (eight sheep in each group) and fed with four diets containing a similar starch content, but from different starch sources, including 100% of starch derived from corn (CS), 33% of starch derived from barley + 67% of starch derived from corn (33 BS), 67% of starch derived from barley + 33% of starch derived from corn (67 BS) and 100% of starch derived from barley (100 BS). The experimental period included a 14 d adaptation period and a 2 d continuous data collection period. The results showed that the molar proportions of acetate, isobutyrate, butyrate and isovalerate and the ratio of acetate to propionate in the 67 BS and 100 BS groups decreased compared with the CS and 33 BS groups (p < 0.001), while the molar proportions of propionate and valerate increased (p < 0.001). The combination of 33% barley starch and 67% corn starch in the diet improved the production of TVFAs (p = 0.007). The OTUs and Shannon indexes of the CS and 33 BS groups were higher than the 67 BS and 100 BS groups (p < 0.001), and the Chao1 and Ace indexes were higher than the 100 BS group (p < 0.05). In addition, the 33 BS group had increased the relative abundances of Bacteroidetes, Prevotella and Ruminococcus and the abundances of Fibrobacter succinogenes, Ruminococcus flavefaciens, Streptococcus bovis, Selenomonas ruminantium and Prevotella brevis relative to the CS group (p < 0.05). These results indicate that the substitution of 33% of the CS with BS did not change the rumen fermentation pattern relative to the CS group, and increased the richness and diversity of the rumen microbes in Hu sheep compared with other two starch substitute groups. [ABSTRACT FROM AUTHOR]

Background: This study investigated greenhouse gas (GHG) emission characteristics of lactating Holstein dairy cows in East China and provided a basis for formulating GHG emission reduction measures. GreenFeed system was used to measure the amount of methane (CH4) and carbon dioxide (CO2) emitted by the cows through respiration. Data from a commercial cow farm were used to observe the effects of parity, body weight, milk yield, and milk component yield on CH4 and CO2 emissions. Results: Mean herd responses throughout the study were as follows: 111 cows completed all experimental processes, while 42 cows were rejected because they were sick or had not visited the GreenFeed system 20 times. On average, lactating days of cows was 138 ± 19.04 d, metabolic weight was 136.5 ± 9.5 kg, parity was 2.8 ± 1.0, dry matter intake (DMI) was 23.1 ± 2.6 kg/d, and milk yield was 38.1 ± 6.9 kg/d. The GreenFeed system revealed that CH4 production (expressed in CO2 equivalent, CO2-eq) was found to be 8304 g/d, C H 4 ( C O 2 − e q) /DMI was 359 g/kg, C H 4 ( C O 2 − e q) /energy-corrected milk (ECM) was 229.5 g/kg, total CO2 production (CH4 production plus CO2 production) was 19,201 g/d, total CO2/DMI was 831 g/kg, and total CO2/ECM was 531 g/kg. The parity and metabolic weight of cows had no significant effect on total CO2 emissions (P > 0.05). Cows with high milk yield, milk fat yield, milk protein yield, and total milk solids yield produced more total CO2 (P < 0.05), but their total CO2 production per kg of ECM was low (P < 0.05). The total CO2/ECM of the medium and high milk yield groups was 17% and 27% lower than that of the low milk yield group, respectively. Conclusions: The parity and body condition had no effect on total CO2 emissions, while the total CO2/ECM was negatively correlated with milk yield, milk fat yield, milk protein yield, and total milk solids yield in lactating Holstein dairy cows. Measurement of total CO2 emissions of dairy cows in the Chinese production system will help establish regional or national GHG inventories and develop mitigation approaches to dairy production regimes. [ABSTRACT FROM AUTHOR]

Reactive oxygen species causes oxidative stress, which oxidizes polyunsaturated fatty acids (PUFAs) to form oxidative metabolites. Sertoli cell is an important cellular metabolism of PUFA in testicular cells, and it regulates the testis development and spermatogenesis. However, the oxylipins generated in testes with different developmental statuses are lacking. In this study, twelve 6-month-old Hu sheep were selected and divided into large testicular group (L) and the small testicular group (S) (n=6). UPLC–MS/MS was conducted to screen oxylipins in the testis, and the total oxylipin and ω-3 PUFA-derived oxylipin contents in the S group were higher. A total of 20 differential oxylipins between the two groups were screened. Among them, the contents of ω-3 PUFA, DHA-derived oxylipins were increased in the S group. The arachidonic acid-derived oxylipin was lower in the S group. The mRNA expression levels of genes related to oxylipin regulation (AKR1B1 , PTGER2 , and PTGDS) were higher in the S group (P < 0.05). In vitro , 200 µM α-linolenic acid alleviated oxidative stress damage to Sertoli cells and improved cell viability by increasing the superoxide dismutase contents and mRNA expression levels of GPX4 and Bcl2. These results indicate that ω-3 PUFA is more susceptible to lipid oxidation in the S group under oxidative stress, which might alleviate the damage of oxidative stress to testis. Moreover, ALA could stimulate the proliferation of Sertoli cells by increasing the capacity of antioxidants. This work may provide a theoretical basis for further studies on the antioxidant properties of the testis for Hu sheep. • UPLC–MS/MS screened 118 oxylipins in the sheep testis. • Ω-3 PUFA is more susceptible to lipid oxidation under oxidative stress in the small testis. • ALA could alleviate oxidative stress damage to Sertoli cells. [ABSTRACT FROM AUTHOR]

Oxidative stress damages the structure of sperm and affects its fertility. Further maturation and storage of spermatozoa occur in the epididymis. Thus, a balanced environment between free-radical production and the antioxidant–enzyme system is important. This study aimed to investigate differences in antioxidative activity in epididymis between high and low fertility levels of Hu sheep. A total of 179 Hu ram lambs aged 56 days were fed up to 6 months in the same environment. After feeding trial, all rams were slaughtered. According to the epididymis weight, cauda sperm concentration, and histo-morphological observation, 18 individuals were selected and divided into low fertility (n = 9) and high fertility (n = 9) groups. The RNA sequencing results showed 1404 differentially expressed genes (DEGs), with 735 genes upregulated and 669 genes downregulated in the high fertility group compared with the low fertility group. Sixteen DEGs related to lipid metabolism and oxidative stress, including GPX3 , GPX5 , LRP1 , PLCB1 , CRISP2 , BSPH1 , and ACSBG1 , were screened by qPCR, and results were correlated well with the sequencing data. Antioxidative indices such as total antioxidant capacity (21.29 ± 5.97 vs. 30.23 ± 4.56 μM/gprot) and glutathione peroxidase (39.75 ± 7.61 vs. 68.84 ± 12.51 U/mgprot) increased in the high fertility group. However, the relative expression of mtDNA and malondialdehyde concentration (0.75 ± 0.15 vs. 0.54 ± 0.10 nmol/mgprot) and the reactive oxygen species (ROS) content significantly decreased (P < 0.05). Therefore, epididymis from high-fertility group had strong antioxidative activity and protected spermatozoa from ROS. [Display omitted] • The diameter of epididymal ducts was increased in epididymis with high fertility of Hu sheep. • RNA-Seq detected differences in epididymis cauda transcriptome. • There were 1404 differentially expressed genes between the high fertility and low fertility groups. • The genes affecting lipid metabolism and sperm maturation were screened out. • Epididymis from high fertility group had strong antioxidative activity, protecting spermatozoa from reactive oxygen species. [ABSTRACT FROM AUTHOR]

Summary: Variations in the Y‐chromosome are usually correlated with male‐specific traits. However, this condition has been described only sporadically, even in human genetics. The present study was conducted to clone the full‐length gene sequence of ovine DEAD‐box helicase 3, Y‐linked (DDX3Y), and investigate the effect of the expression and variation of DDX3Y on the reproductive traits of Hu sheep. Consequently, we identified the full coding sequence and genomic sequence of ovine DDX3Y. Quantitative PCR (qPCR) analysis showed that ovine DDX3Y was highly expressed in testis, and the expression level increased during testicular development. Furthermore, individuals with larger testis at 6 months expressed significantly more DDX3Y mRNA in the testis than individuals with smaller testis. Notably, a novel SNP (g. 12657 C>A) in the 3′ untranslated region was identified in Hu sheep and Tan sheep according to the investigation of the full DDX3Y genomic sequence of 1069 individuals from nine sheep breeds. Association analysis revealed that the SNP was significantly related to testis size in Hu sheep. Meanwhile, Hu rams with the derived C allele showed significantly higher expression levels of DDX3Y in testis than those with the ancestral A allele. In addition, data mining in a previous study showed that the C allele cosegregated with the globally major Y‐chromosomal haplogroups y‐HA and y‐HC, and the A allele is found in all rams with haplogroups y‐HB1, y‐HB2 and y‐HD. This study suggests that the association of the Y‐chromosomal haplogroups with testis size in Hu sheep can be extrapolated to the sheep population worldwide. [ABSTRACT FROM AUTHOR]