A sheep pangenome reveals the spectrum of structural variations and their effects on tail phenotypes.

Structural variations (SVs) are a major contributor to genetic diversity and phenotypic variations, but their prevalence and functions in domestic animals are largely unexplored. Here we generated high-quality genome assemblies for 15 individuals from genetically diverse sheep breeds using Pacific Biosciences (PacBio) high-fidelity sequencing, discovering 130.3 Mb nonreference sequences, from which 588 genes were annotated. A total of 149,158 biallelic insertions/deletions, 6531 divergent alleles, and 14,707 multiallelic variations with precise breakpoints were discovered. The SV spectrum is characterized by an excess of derived insertions compared to deletions (94,422 vs. 33,571), suggesting recent active LINE expansions in sheep. Nearly half of the SVs display low to moderate linkage disequilibrium with surrounding single-nucleotide polymorphisms (SNPs) and most SVs cannot be tagged by SNP probes from the widely used ovine 50K SNP chip. We identified 865 population-stratified SVs including 122 SVs possibly derived in the domestication process among 690 individuals from sheep breeds worldwide. A novel 168-bp insertion in the 5' untranslated region (5' UTR) of HOXB13 is found at high frequency in long-tailed sheep. Further genome-wide association study and gene expression analyses suggest that this mutation is causative for the long-tail trait. In summary, we have developed a panel of high-quality de novo assemblies and present a catalog of structural variations in sheep. Our data capture abundant candidate functional variations that were previously unexplored and provide a fundamental resource for understanding trait biology in sheep.

Deep Sequencing of the Rat MCAO Cortexes Reveals Crucial circRNAs Involved in Early Stroke Events and Their Regulatory Networks.

Circular RNAs (circRNAs) are highly enriched in the central nervous system and significantly involved in a range of brain-related physiological and pathological processes. Ischemic stroke is a complex disorder caused by multiple factors; however, whether brain-derived circRNAs participate in the complex regulatory networks involved in stroke pathogenesis remains unknown. Here, we successfully constructed a cerebral ischemia-injury model of middle cerebral artery occlusion (MCAO) in male Sprague-Dawley rats. Preliminary qualitative and quantitative analyses of poststroke cortical circRNAs were performed through deep sequencing, and RT-PCR and qRT-PCR were used for validation. Of the 24,858 circRNAs expressed in the rat cerebral cortex, 294 circRNAs were differentially expressed in the ipsilateral cerebral cortex between the MCAO and sham rat groups. Cluster, GO, and KEGG analyses showed enrichments of these circRNAs and their host genes in numerous biological processes and pathways closely related to stroke. We selected 106 of the 294 circRNAs and constructed a circRNA-miRNA-mRNA interaction network comprising 577 sponge miRNAs and 696 target mRNAs. In total, 15 key potential circRNAs were predicted to be involved in the posttranscriptional regulation of a series of downstream target genes, which are widely implicated in poststroke processes, such as oxidative stress, apoptosis, inflammatory response, and nerve regeneration, through the competing endogenous RNA mechanism. Thus, circRNAs appear to be involved in multilevel actions that regulate the vast network of multiple mechanisms and events that occur after a stroke. These results provide novel insights into the complex pathophysiological mechanisms of stroke.

Exploring the roles of fecundity-related long non-coding RNAs and mRNAs in the adrenal glands of small-tailed Han Sheep.

BACKGROUND: Long non-coding RNAs (lncRNAs) can play important roles in uterine and ovarian functions. However, little researches have been done on the role of lncRNAs in the adrenal gland of sheep. Herein, RNA sequencing was used to compare and analyze gene expressions in adrenal tissues between follicular phases and luteal phases in FecBBB (MM) and FecB++ (WW) sheep, respectively, and differentially expressed lncRNAs and genes associated with reproduction were identified. RESULTS: In MM sheep, 38 lncRNAs and 545 mRNAs were differentially expressed in the adrenal gland between the luteal and follicular phases; In WW sheep, 513 differentially expressed lncRNAs and 2481 mRNAs were identified. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses indicated that differentially expressed lncRNAs and their target genes are mainly involved in the circadian rhythm, the mitogen activated protein kinase, thyroid, ovarian steroidogenesis and transforming growth factor beta signaling pathways. Differentially expressed lncRNAs can regulate reproduction by modulating genes involved in these signaling pathways and biological processes. Specifically, XLOC_254761, XLOC_357966, 105,614,839 and XLOC_212877 targeting CREB1, PER3, SMAD1 and TGFBR2, respectively, appear to play key regulatory roles. CONCLUSION: These results broaden our understanding of lncRNAs in adrenal gland of sheep and provide new insights into the molecular mechanisms underlying sheep reproduction.

Identification of Prolificacy-Related Differentially Expressed Proteins from Sheep (Ovis aries) Hypothalamus by Comparative Proteomics.

Reproduction, as a physiologically complex process, can significantly affect the development of the sheep industry. However, a lack of overall understanding to sheep fecundity has long blocked the progress in sheep breeding and husbandry. In the present study, the aim is to identify differentially expressed proteins (DEPs) from hypothalamus in sheep without FecB mutation in two comparison groups: polytocous (PF) versus monotocous (MF) sheep at follicular phase and polytocous (PL) versus monotocous (ML) sheep at luteal phase. Totally 5058 proteins are identified in sheep hypothalamus, where 22 in PF versus MF, and 39 proteins in PL versus ML are differentially expressed, respectively. A functional analysis is then conducted including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis to reveal the potential roles of these DEPs. The proteins ENSOARP00000020097, ENSOARP00000006714, growth hormone (GH), histone deacetylase 4 (HDAC4), and 5'-3' exoribonuclease 2 (XRN2) in PF versus MF, and bcl-2-associated athanogene 4 (BAG4), insulin-like growth factor-1 receptor (IGF1R), hydroxysteroid 11-beta dehydrogenase 1 (HSD11B1), and transthyretin (TTR) in PL versus ML appear to modulate reproduction, presumably by influencing the activities of gonadotropin-releasing hormone (GnRH). This study provides an alternative method to identify DEPs associated with sheep prolificacy from the hypothalamus. The mass spectrometry data are available via ProteomeXchange with identifier PXD013822.

Effects of active immunization with newly modified GnRH peptides on spermatogenesis and production performance of Holstein bulls.

Immunocastration via vaccination against gonadotropin-releasing hormone (GnRH) is an effective alternative to surgical castration in livestock. In this study, male mice were immunized with eight GnRH peptide derivatives. Two, which exhibited highly significant effects in mice, and one which exhibited the least significant effects were selected for active immunization of 13-month-old bulls. The effects of these GnRH vaccines on sexual development and meat quality in bulls were evaluated by examining testis length, serum hormone and GnRH antibody concentrations, observation of sexual behavior and testicular tissue sections, and evaluation of meat quality indexes. The results indicated that anti-GnRH titers increased rapidly (P < 0.05) and serum follicle stimulating hormone, luteinizing hormone, and testosterone concentrations decreased sharply after booster immunization (P < 0.05), while testis volumes were lower (P < 0.01), testicular growth was arrested and spermatogenesis inhibited in group C GnRH-treated versus control bull groups. Meat quality was not significantly different in immunocastrates relative to bulls in the control group. Our collective results provide a scientific basis to further clarify the mechanisms underlying GnRH-mediated regulation of livestock reproduction, and contribute to the development of an efficient, safe, and reversible immune castration vaccine.

Comparative Proteome Analysis Reveals Lipid Metabolism-Related Protein Networks in Response to Rump Fat Mobilization.

Altay is a typical fat-tailed sheep breed displaying the unique ability to rapidly mobilize fat, which is vital for maintaining a normal metabolism that facilitates its survival in lengthy winter conditions. However, the physiological, biochemical, and molecular mechanisms underlying fat mobilization remain to be elucidated. In this study, the monitoring of rump fat adipocyte sizes disclosed a positive correlation between cell size and fat deposition ability. In addition, we subjected sheep to persistent starvation to imitate the conditions that trigger rump fat mobilization and screened 112 differentially expressed proteins using the isobaric peptide labeling approach. Notably, increased secretion of leptin and adiponectin activated the key fat mobilization signaling pathways under persistent starvation conditions. Furthermore, the upregulation of resistin (RETN), heat-shock protein 72 (HSP72), and complement factor D (CFD) promoted lipolysis, whereas the downregulation of cell death-inducing DFFA-like effector C (CIDEC) inhibited lipid droplet fusion, and the increase in HSP72 and apolipoprotein AI (Apo-AI) levels activated the body's stress mechanisms. The synergistic actions of the above hormones, genes, and signaling pathways form a molecular network that functions in improving the adaptability of Altay sheep to extreme environments. Our findings provide a reference for elucidating the complex molecular mechanisms underlying rump fat mobilization.

[Molecular cloning and tissue expression of the CCNG1 gene in sheep].

The CCNG1 gene encodes cyclin G1, which is an important cell cycle regulator and has been reported to be involved in reproductive biological processes, such as oocyte maturation and granule cell proliferation in mammals. But the study of CCNG1 in sheep has been rarely reported. To examine the effects of CCNG1 on estrous control and seasonal breeding in sheep, we first cloned and characterized the expression level of the sheep CCNG1 gene. Then by Real-time PCR, we detected and analyzed the expressions of CCNG1 gene at mRNA levels in the hypothalamus-pituitary-ovary (HPO) axis in different stages of an estrous cycle in Duo Lang sheep (non-seasonal breeding) and Merino sheep (seasonal breeding). The results showed that the open reading frame of the sheep CCNG1 gene is 885 bp in length and encodes 294 amino acids. Bioinformatic analysis indicated that the secondary structure of the sheep CCNG1 protein contained multiple phosphorylation sites and some Protein Kinase C phosphorylation sites. CCNG1 mRNA was identified in all tissues tested, with the levels in ovary and kidney higher than others. The expression profiles of CCNG1 in the HPO axis in different stages of an estrous cycle were similar in different sheep breeds: the expression levels of CCNG1 in the ovary, uterus, pineal gland and pituitary gland all peaked in the estrus phase. But there were significant differences for expression change extent of CCNG1 in ovaries in the oestrus and metestrus phase between different sheep breeds. The results suggested that CCNG1 probably participated in the regulation of estrous behavior and seasonal reproduction through controling the growth and development of follicles in sheep.

[Analysis of FABP4 expression pattern in rump fat deposition and metabolism of Altay sheep].

FABP4 (Fatty acid binding protein 4) is a hot candidate gene in fat deposition and lipid metabolism and participates in the transport and metabolism of intracellular free fatty acids. We aim to study the role of FABP4 in fat deposition and metabolism of the rump fat in Altay sheep. In this study, bioinformatics method was used to analyze the protein sequence homology among 10 species, and RT-PCR was employed to detect FABP4 tissue profiling of Altay sheep. An animal model simulating the rump fat deposition and metabolism of Altay sheep was established by continuous starvation, and qPCR and iTRAQ (isobaric tags for relative and absolute quantitation) were used to detecte FABP4 mRNA and protein expression changes in the control and continuous starvation groups, respectively. Sequence analysis showed that FABP4 protein sequence is highly conserved among species, suggesting an important biological function during evolution for FABP4. The RT-PCR result confirmed that FABP4 mRNA was highly expressed in intestinal and rump fat, suggesting that FABP4 plays an important physiological role in fat tissues. We did not find significant differences in FABP4 mRNA and protein between control and continuous starvation groups (P>0.05), which indicates that FABP4 may not be the key gene in fat deposition and metabolism in Altay sheep.The results above lay a foundation for further studies of FABP4 in rump or tail fat.

Alginate Oligosaccharide Alleviates Lipopolysaccharide-Induced Apoptosis and Inflammatory Response of Rumen Epithelial Cells through NF-κB Signaling Pathway.

AOS alleviates inflammatory responses; however, whether it exerts an effect on the rumen or regulates rumen inflammatory reaction remains unknown. In this study, firstly, the ovine ruminal epithelial cells (ORECs) were treated with 0, 200, 400, 600, and 800 µg/mL AOS, hoping to explore whether AOS hurt cell health. The results showed that compared with the AOS-0 group, the AOS-400 group could significantly increase (p < 0.05) cell viability, reduce (p < 0.05) reactive oxygen species (ROS) and interleukin (IL)-6 content, and have no adverse effect on cells. Secondly, we used LPS to construct an in vitro inflammatory model of rumen epithelial cells and then explored the protective role of AOS on rumen epithelial cells. The study was divided into three groups: the control group (CON), LPS, and LPS + AOS. The results demonstrated that the LPS + AOS group significantly increased the cell viability and reduced the ROS level in comparison with the LPS group (p < 0.05). Pretreatment with AOS also repressed (p < 0.05) the secretion of IL-1ß, IL-6, IL-8, and immunoglobulin (Ig)A from ORECs in the culture medium following LPS. In terms of tight junction (TJ) proteins, AOS treatment also significantly increased (p < 0.05) the zonula occludens 1 (ZO-1) and Occludin expression. The apoptosis rate, Caspase3, Caspase9, BAD, and BCL-2/BAX were decreased (p < 0.05) after AOS treatment, and the expression of BCL-2 was increased (p < 0.05). In addition, the expressions of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and nuclear factor-κB (NF-κB) were inhibited (p < 0.05) with the addition of AOS. At the protein level, pretreatment of AOS decreased (p < 0.05) the expression of MyD88 and the phosphorylation level of inhibitor κB α (IκBα) after the LPS challenge. Taken together, our results indicated that AOS could alleviate the LPS-induced apoptosis and inflammatory response of rumen epithelial cells through the NF-κB signaling pathway, which may be a promising strategy for treating apoptosis and inflammation in sheep breeding.

Replacing alfalfa hay with amaranth hay: effects on production performance, rumen fermentation, nutrient digestibility and antioxidant ability in dairy cow.

OBJECTIVE: The aim of this research was to explore the effects of dietary substitution of alfalfa hay by amaranth hay on production performance, rumen fermentation, nutrient digestibility, serum biochemical parameters and antioxidant ability in dairy cows. METHODS: A total of 45 healthy Holstein cows with same parity and similar milk yield and body weight were randomly divided into 3 groups: control diet without amaranth hay (CON) or 50% and 100% alfalfa hay replaced by an equal amount of amaranth hay (dry matter basis, AH1 and AH2, respectively). All the cows were fed regularly 3 times a day at 06:30, 14:30, and 22:30 and had free access to water. The experiment lasted for 60 d. RESULTS: The dry matter intake of CON and AH1 groups was higher (p<0.05) than that of AH2 group. Compared with AH1 group, the milk yield of AH2 group was reduced (p<0.05). Moreover, dietary substitution of alfalfa hay by amaranth hay increased (p<0.05) milk fat, ammonia nitrogen and acetate concentrations. However, the crude protein digestibility of AH2 group was lower (p<0.05) than that of CON group, while an opposite tendency of serum urea nitrogen was found between two groups. The neutral detergent fiber digestibility of AH1 group was increased (p<0.05) when compared to AH2 group. Amaranth hay treatment increased (p<0.05) the serum concentration of glutathione peroxidase in dairy cows. Compared with CON group, the malonaldehyde activity of AH1 group was decreased (p<0.05). CONCLUSION: Dietary replacing alfalfa hay with amaranth hay (50% ratio) in dairy cows did not affect production performance but improved their antioxidant ability.

Effect of tea polyphenols on intestinal barrier and immune function in weaned lambs.

Introduction: The purpose of this study was to explore the effects of tea polyphenols on growth performance, cytokine content, intestinal antioxidant status and intestinal barrier function of lambs, in order to provide reference for intestinal health of ruminants. Methods: Thirty weaned lambs (average initial weight 9.32 ± 1.72 kg) were randomly divided into five groups with six lambs in each group. The control group did not add anything but the basic diet mainly composed of Pennisetum and Corn, and the other four groups added 2, 4, 6 g/kg tea polyphenols and 50 mg/kg chlortetracycline to the basic diet, respectively. The experiment lasted for 42 days. Results: Dietary tea polyphenols improved the growth and stress response and reduced intestinal permeability of lambs (p > 0.05), while CTC did not affect the final lamb weight (p > 0.05). Both tea phenols and CTC significantly reduced inflammatory factors and enhanced the immune system (p > 0.05). Dietary tea polyphenols increased villus height, villus height/crypt depth, secretory immunoglobulin A (p > 0.05), and antioxidant enzymes, while decreasing MDA and apoptosis in the intestine (p > 0.05). However, compared with other groups, the content of T-AOC in jejunum did not change significantly (p > 0.05). Tea polyphenols also increased claudin-1 levels in the duodenum, jejunum, and ileum more than CTC (p > 0.05). CTC had a limited effect on the mRNA expression of Occludin and ZO-1, while tea polyphenols increased these in both the duodenum and ileum (p > 0.05). Conclusion: This study demonstrated that tea polyphenols can effectively improve the intestinal barrier of weaned lambs, and that they have anti-inflammatory and antioxidant effects similar to those of antibiotics. Thus, tea polyphenols could be used to replace antibiotics in ensuring safety of livestock products and in achieving the sustainable development of modern animal husbandry.

Compendium of 5810 genomes of sheep and goat gut microbiomes provides new insights into the glycan and mucin utilization.

BACKGROUND: Ruminant gut microbiota are critical in ecological adaptation, evolution, and nutrition utilization because it regulates energy metabolism, promotes nutrient absorption, and improves immune function. To study the functional roles of key gut microbiota in sheep and goats, it is essential to construct reference microbial gene catalogs and high-quality microbial genomes database. RESULTS: A total of 320 fecal samples were collected from 21 different sheep and goat breeds, originating from 32 distinct farms. Metagenomic deep sequencing and binning assembly were utilized to construct a comprehensive microbial genome information database for the gut microbiota. We successfully generated the largest reference gene catalogs for gut microbiota in sheep and goats, containing over 162 million and 82 million nonredundant predicted genes, respectively, with 49 million shared nonredundant predicted genes and 1138 shared species. We found that the rearing environment has a greater impact on microbial composition and function than the host's species effect. Through subsequent assembly, we obtained 5810 medium- and high-quality metagenome-assembled genomes (MAGs), out of which 2661 were yet unidentified species. Among these MAGs, we identified 91 bacterial taxa that specifically colonize the sheep gut, which encode polysaccharide utilization loci for glycan and mucin degradation. CONCLUSIONS: By shedding light on the co-symbiotic microbial communities in the gut of small ruminants, our study significantly enhances the understanding of their nutrient degradation and disease susceptibility. Our findings emphasize the vast potential of untapped resources in functional bacterial species within ruminants, further expanding our knowledge of how the ruminant gut microbiota recognizes and processes glycan and mucins. Video Abstract.

[Relative quantification of mRNA transcription of Cry1 in different tissues of sheep in oestrous cycle by real-time quantitative PCR].

Studies have shown that clock gene Cry1 may have important roles in the endocrine process of seasonal reproduction in mammals. In this study, Duolang sheep (non-seasonal reproduction sheep breed) and Chinese Merino (seasonal reproduction sheep breed) were used to determine the expression change of Cry1 in hypothalamus-pituitary-ovary axis in different stage of estrous cycle by quantitative real-time PCR. The results showed that the Cry1 mRNA was expressed in all tested tissues, in which the expression levels in pineal gland and thyroid gland were higher than in other tissues. As far as different sheep breeds were concerned, the tissue expression profiles of Cry1 at different stage of estrous cycle were broadly similar. Besides hypothalamus, the expression levels of Cry1 in ovary, uterus, pineal gland, pituitary gland, and thyroid gland were all reached to peak in proestrus. The differences of expression change extent for Cry1 in vary, uterus, pineal gland, and pituitary gland in proestrus and oestrus were significant between different sheep breeds. The results suggested that Cry1 may play roles in switching on the estrus and seasonal reproduction.

[Correlation analysis between polymorphism of the 59383635th locus on X chromosome and fat-tail trait in sheep].

Fat tail or fat rump is one of essential traits for surviving in harsh environments, and the mechanism of fat deposition and its inheritable characters in sheep are still unclear. Therefore, the 59383635th locus on X chromosome in our unpublished chip data was chosen as candidate SNP, PCR-SSCP method was used to detect genotypes in five sheep breeds which have extreme differences in tail types (Altay, Small Tail Han Sheep, Hu, Chinese Merino and Suffolk), and the mathematical model was employed to analyze the correlation between the polymorphism and the trait of fat tail or fat rump. The results in this study showed that the high frequency of allele T exists in Altay flock, and the frequency of allele C appears to be particularly high in the thin tail sheep breeds. The result of mathematical model showed that the ratio of T/C increased exponentially with the increase of phenotype score. These results suggest that there is a big difference in the SNP distribution between fat tail (rump) and thin tail sheep populations, and the SNP can be used as an ideal molecular marker in high-fat or low-fat sheep breeding. However, the biological function of the SNP remains to be further studied.

[Detection and analysis of polymorphisms of 59571364 and 59912586 loci on X chromosome in fat-tail and thin-tail sheep flocks].

In order to analyze the correlation of tail fat deposition and two SNP loci on Ovis arise chromosome X and provide a theoretical basis for using molecular assisted selection technology in further low-fat sheep breeding, the breeds with extreme differences in tail types (Altay, Small Tail Han Sheep, Hu, Chinese Merino and Suffolk) were used to detect the polymorphisms of two SNP loci on X chromosome and the haplotypes with PCR-RFLP method. The results showed that the TT genotype at 59571364 locus and GG genotype at 59912586 locus were preponderant genotypes in thin-tailed Chinese Merino and Suffolk sheep flocks, while the percentage of the two genotypes in fat-tailed (fat-rmup) Altay and Hu flocks is less than 2%. Haplotype analysis showed that CA haplotype is the main haplotype, the percentage of CA is up to 55%, and the percentage of CA and TA haplotypes together was 88.33% in Altay sheep flock. These results suggest that there are great differences in the SNP distribution of the 59571364 and 59912586 loci among different tail-typed sheep flocks, which can be used as molecular markers in high or low fat sheep breeding.

Silage additives improve fermentation quality, aerobic stability and rumen degradation in mixed silage composed of amaranth and corn straw.

The objective of this research was to investigate effects of different additives on the fermentation quality, aerobic stability and rumen degradation of mixed silage composed of amaranth and corn straw. The mixture ratio of amaranth to corn straw was 78%: 22%. Three additives were selected in this study and five groups were as follows: control group (CON, without additive), lactic acid bacteria group (LAB, 5 mg/kg, Lactobacillus plantarum ≥ 1.6×1010 CFU/g and L. buchneri ≥ 4.0×109 CFU/g), glucose group (GLU, 30 g/kg), cellulase group (CEL, 2 mg/kg) and lactic acid bacteria, glucose and cellulase group (LGC, added at the same levels as in individual group). The period of ensiling was 60 days. Fermentation quality, chemical composition and aerobic stability of mixed silage were analyzed. Four cows with permanent ruminal fistula were selected as experimental animals. Nylon bag technique was used to study rumen degradation characteristic of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) of mixed silage. Compared with CON group, the addition of different silage additives could improve mixed silage quality of amaranth and corn straw to some extent. Combining three additives significantly increased (P < 0.05) the DM, CP and lactic acid contents, whereas decreased (P < 0.05) the ADF and NDF contents as well as pH and ammonia nitrogen/total nitrogen. Moreover, the aerobic stability and rumen degradation of DM, CP and NDF were significantly improved (P < 0.05) in LGC group when compared to other groups. In conclusion, the combined addition of lactic acid bacteria, glucose and cellulase increased DM, CP and lactic acid contents as well as lactic acid bacteria count, decreased NDF and ADF contents and aerobic bacteria and mold counts, improved aerobic stability and rumen degradation of amaranth and corn straw mixed silage.

Grape Seed Extract as a Feed Additive Improves the Growth Performance, Ruminal Fermentation and Immunity of Weaned Beef Calves.

The purpose of this research was to evaluate effects of grape seed extract (Gse) supplementation on the growth performance; ruminal fermentation; nutrient digestibility; and serum biochemical, antioxidative, and immune parameters of weaned beef calves. A total of 30 Simmental crossbred male calves with similar age and body weight were randomly allocated to two groups: a control group with no Gse (CON) and a Gse supplementation group (GSE) (4 g/d Gse per animal). The results show that, compared with the CON group, the average daily gain significantly increased (p = 0.043) in the GSE group. The ruminal contents of microbial protein and butyrate in GSE group were higher (p < 0.05) than those in the CON group. Additionally, calves fed Gse displayed increased (p < 0.05) dry matter and neutral detergent fiber digestibility. Moreover, the serum concentrations of triglyceride, catalase, superoxide dismutase, immunoglobulin G and immunoglobulin M were higher (p < 0.05) in the GSE group than those in the CON group. However, opposite tendencies of non-esterified fatty acid, malondialdehyde, tumor necrosis factor-α and interleukin-6 were found between the two groups. Overall, the supplementation of Gse can improve ruminal fermentation, nutrient digestibility, antioxidant ability, and immunity, as well as promoting the healthy growth of weaned cross-breed beef calves.

[Cloning and expression in follicle anagen of ILK gene in sheep].

Integrin linked kinase (ILK) is a scaffold protein, which plays important roles in hair follicle development. The cDNA sequence of novel ILK gene in sheep was cloned by PCR method and analyzed by bioinformatics. Tissue expression profiling in eight tissues and temporal profiling at different wool follicle anagen stages in skin was analyzed. The results showed that the whole open reading frame (ORF) of ILK gene was 1 359 bp in length, which encoded 452 amino acids. Bioinformatic analysis indicated that the secondary structure of ILK gene was mainly made up of three ankyrin repeats and a kinase domain, and there were multiple phosphorylation and Protein Kinase C sites in this gene. The RT-PCR result confirmed that ILK mRNA was expressed in heart, liver, spleen, lung, skeletal muscle, skin, and small intestine, and the expression level was much higher in skin, spleen, and liver than others. The q-PCR analysis demonstrated that the ex-pression level of ILK was significantly increased from March to May (early follicle anagen initiation) in both sheep breeds, Chinese Merino and Kazakh sheep, and there were certain differences from June to October between the two breeds. The above results indicated that ILK gene may play key roles in regulating secondary follicle growth.

The effects of fucoidan as a dairy substitute on diarrhea rate and intestinal barrier function of the large intestine in weaned lambs.

This paper explores the effects of fucoidan on the frequency of diarrhea, colon morphology, colon antioxidant status, cytokine content, short-chain fatty acids, and microflora of cecal contents in early weaned lambs in order to provide a reference for the intestinal health of young ruminants. Fucoidan is a natural active polysaccharide extracted from kelp and other large brown algae. It has many biological effects, such as improving immunity, nourishing the stomach and intestines, and anti-tumor properties. This study investigated the effects of fucoidan supplementation in milk replacer on the large intestine's ability to act as an intestinal barrier in weaned lambs. With six duplicate pens and one lamb per pen, a total of 24 weaned lambs (average starting body weight of 7.32 ± 0.37 kg) were randomly assigned to one of four milk replacer treatments. Four concentrations of fucoidan supplementation (0, 0.1, 0.3, and 0.6% dry matter intake) were employed to investigate the effects of fucoidan on cecal fermentation and colon microbial organization. The test period lasted 37 days (1 week before the test and 1 month after the test), and lamb cecal contents and colon organization were collected for examination. In addition, the fecal status of all lambs was observed and recorded daily, allowing us to calculate the incidence of diarrhea in weaned lambs. The findings demonstrated that fucoidan may significantly increase the concentration of short-chain fatty acids (propionic acid and butyric acid) in the cecal digesta of weaned lambs. In weaned lambs, 16S rDNA testing showed that fucoidan at 0.3-0.6% (dry matter intake) was beneficial for boosting the variety of the intestinal bacteria and modifying the relative abundance of a few bacterial strains. In addition, fucoidan enhanced colon antioxidant and immune functions and decreased the diarrhea rate to relieve weaning stress. This result demonstrates that milk replacer supplementation with fucoidan contributes to the improvement in the large intestinal health of weaned lambs.

Effects of Nano-Selenium on Antioxidant Capacity in Se-Deprived Tibetan Gazelle (Procapra picticaudata) in the Qinghai-Tibet Plateau.

Tibetan gazelle (Procapra picticaudata) is an endangered ungulate in the Qinghai-Tibet Plateau, China. This study aimed to determine the influence of nano-Se on antioxidant system in Se-deprived P. picticaudata. We analyzed contents of mineral elements in soil, forage, and animal tissue. Blood parameters and antioxidant indexes were also determined. The results showed that Se concentrations in the soil and forage from affected pasture were significantly lower than those in healthy area (P < 0.01). Se concentrations in blood and hair from affected P. picticaudata were also significantly lower than those in healthy animals (P < 0.01). Meanwhile, the levels of Hb, RBC, and PCV in affected gazelle were significantly lower than those in healthy animal (P < 0.01). The activities of AST, ALT, LDH, CK, and UA content in affected animal were significantly lower than those in healthy gazelles (P < 0.01). The levels of SOD, GSH-Px, CAT, and T-AOC in serum were significantly lower and the MDA content was significantly higher in affected compared with healthy gazelle (P < 0.01). Affected P. picticaudata were treated orally with nano-Se, Se concentration in blood significantly increased and serum antioxidant indexes greatly returned to within the healthy range. Consequently, nano-Se could not only markedly increase the Se content in blood in Se-deprived P. picticaudata but also much improves the antioxidant capacity.