Supplementation of Lycium barbarum residue increases the growth rate of Tan sheep by enhancing their feed intake and regulating their rumen microbiome and metabolome

Lycium barbarum residue(LBR),a by-product of L.barbarum processing,is packed with bioactive components and can be potentially utilized as a feed additive in animal husbandry.However,the fundamental understanding of its effectiveness on livestock animals is still lacking,particularly in ruminants.To explore the effects of LBR on the growth performance,rumen fermentation parameters,ruminal microbes and metabolites of Tan sheep,sixteen fattening rams(aged 4 mon)were fed a basal diet(CON,n=8)or a basal diet supplemented with 5%LBR(LBR,n=8).The experiment lasted for 70 d,with 10 d adaptation period and 60 d treatment period.The results showed that the LBR enhanced the average daily feed intake,average daily gain(P<0.05),and ruminal total volatile fatty acids(P<0.01)while decreasing ammonia-nitrogen concentration and rumen pH value(P<0.05).Additionally,the LBR improved the relative abundances of Prevotella,Succiniclasticum,Ruminococcus,Coprococcus,Selenomonas,and Butyrivibrio(P<0.05)and reduced the relative abundances of Oscillospira and Succinivibrio(P<0.05).The LBR altered the ruminal metabolome(P<0.01)by increasing the abundances of ruminal metabolites involved in amino acids(e.g.,L-proline,L-phenylalanine,L-lysine,and L-tyrosine),pyrimidine metabolism(e.g.,uridine,uracil,and thymidine),and microbial protein synthesis(e.g.,xanthine and hypoxanthine).In conclusion,LBR had positive effects on the growth rate of Tan sheep as well as on rumen fermentation parameters,rumen microbiome and rumen metabolome.

Effects of Antimicrobial Peptides on Production,Slaughter Performance and Blood Routine of Tan Sheep

[Objectives]This study was conducted to analyze effects of antimicrobial peptides added to the diet of Tan sheep on their production, slaughter performance and blood composition. [Methods] Ninety two four-month-old Tan sheep were randomly divided into two treatment groups according to their body weight, 46 in each group. The control check group(CK) was feed with conventional diet, and the experimental group was fed with the addition of antimicrobial peptide on the basis of the conventional diet, with the added amount of 2.5 g/sheep per day. The experimental period was 60 d. [Results] The incidence rate was 75.06% lower in the experimental group than in the CK. The average daily weight gain per sheep was 11.27% higher in the experimental group than in the CK(P<0.05). The feed conversion ratio was 8.45% lower in the experimental group than in the CK(P<0.05). The average daily gross profit per sheep was 12.12% higher in the experimental group than in the CK. For slaughter performance, the data difference of each item was not significant. The PH at 45 min and 24 h after slaughter was within the normal range of fresh mutton. The cooked meat percentage and water loss rate showed no significant differences(P>0.05). The marbling ranged from 2.45 to 2.50, indicating that the fat content was moderate, and the difference between groups was not significant(P>0.05). The flesh color ranged from 3.00 to 3.15, between light red and bright red, belonging to the normal color of mutton, and the difference between groups was not significant(P>0.05). The shear force was between 2.50 and 2.65, without a significant difference between groups(P>0.05). The white blood cells, lymphocytes and platelets in the experimental group were lower than those in the CK(P<0.01). The erythrocytes and hemoglobin in the erythrocyte group were higher than those in the CK(P<0.05). The neutrophils and monocytes in the experimental group were lower than those in the CK(P<0.05). [Conclusions] This study provides a technical basis for the rational use of antimicrobial peptides and their application in ruminants.

The Effects of Nutrition Lick Brick and Salt Brick as Supplement Feed on the Production Performance of Tan Sheep

In the paper, the effects of nutrition lick brick and salt brick as supple-ment feed on the performance of Tan sheep were studied. 36 fattening Tan sheep with the average weight of about 23 kg were chosen, they were randomly divided into 3 groups according to the weight, and each group had 12 sheep. Control group was fed with normal daily ration, experimental group 1 was fed with normal daily ration＋nutrition lick brick, experimental group 2 was fed with normal daily ration＋salt brick, the experimental period was 60 d. The results showed that 1） compared with CK, daily gain of group 1 improved 17.09%（P〈0.01）, feed conversion ratio reduced 11.69%（P〈0.05）, and average gross profit per sheep improved 51.65 yuan; daily gain of group 2 improved 7.03%（P〈0.05）, feed conversion ratio reduced 4.06%（P〉0.05）, and average gross profit per sheep improved 19.78 yuan; 2） compared with group 2, daily gain of group 1 improved 9.40%（P〈0.05）, feed conversion ratio re-duced 7.96%（P〈0.05）, and average gross profit per sheep improved 31.87 yuan;these indicated that nutrition lick brick and salt brick as supplement feed for Tan sheep both can improve the production performance of Tan sheep, and the supple-mentary feeding effect of nutrition lick brick was better.

In vitro Maturation of Tan Sheep Oocytes

[Objective] This study aimed to investigate the appropriate concentrations of follicle-stimulating hormone（FSH）, luteotropic hormone（LH） and estrodiol（E2） during in vitro maturation of Tan sheep oocytes. [Method] Tan sheep oocytes were divided into five groups for in vitro maturation culture： control group, FSH group（10,50, 100, 200 and 300 μg/ml FSH, respectively）, LH group（5, 10, 20, 50 and 100μg/ml LH, respectively）, E2group（5, 10, 25, 50 and 100 μg/ml E2, respectively）, and FSH ＋ LH group（100 μg/ml FSH ＋ 20 μg/ml LH）. The releasing rate of first polar bodies was analyzed. [Result] The maturation rate of Tan sheep oocytes in 100 μg/ml FSH ＋ 20 μg/ml LH group reached the highest（64.64%）, which was significantly higher than that in other four groups（P〈0.05）; among different FSH concentrations,100 μg/ml FSH was superior to other four concentrations and the control group, exhibiting significant differences（P〈0.05）; among different LH concentrations, 20 μg/ml LH was superior to other four concentrations and the control group, exhibiting significant differences（P〈0.05）; among different E2 concentrations, 50 μg/ml E2 was superior to other four concentrations and the control group, exhibiting significant differences（P〈0.05）. [Conclusion] Under the experimental conditions, 100 μg/ml FSH ＋20 μg/ml LH was the most appropriate hormone combination for in vitro maturation of Tan sheep oocytes.

The Levels of Genetic Differentiation of Small-Tailed Han Sheep and Tan Sheep Populations Using Structural Loci

Using the method of ＂random sampling in typical colonies of the central area of the habitat＂ and several electrophoresis techniques, the variations of 17 structural loci encoding blood proteins in 60 Small-Tailed Han sheep and 73 Tan sheep were examined and compared with those of 14 other sheep populations in China and other countries to investigate their levels of genetic differentiation. The average heterozygosities of Small-Tailed Han sheep and Tan sheep were 0.2360 and 0.2587, respectively. The average polymorphic information content values were 0.1974 and 0.2102, respectively. The average effective numbers of alleles were 1.5723 and 1.5751, respectively. The coefficients of gene differentiation in the four groups （including 4, 6, 13, and 16 sheep populations, respectively） were 0.049323, 0.059987, 0.1728, and 0.201256, respectively, indicating that the degree of gene differentiation at the structural loci was the least in Hu sheep, Tong sheep, Small-Tailed Hart sheep, and Tan sheep; followed by the above-mentioned four sheep populations and two Mongolian sheep populations; and was the highest in sheep populations belonging to the Mongolian sheep group, South Asian sheep, and European sheep. The earlier researchers＇ conclusions that both Small-Tailed Han sheep and Tan sheep evolved from Mongolian sheep were further verified by the results of this study. Hu sheep, Tong sheep, Small-Tailed Han sheep, and Tan sheep were decreasingly affected by the bloodline of Mongolian sheep to different degrees. The relationships among sheep populations were not closely related to the geographical distances among sheep populations.

Dynamic changes of genomic methylation profiles at different growth stages in Chinese Tan sheep

Background:Tan sheep,an important local sheep breed in China,is famous for their fur quality.One-month-old Tan sheep have white,curly hair with beautiful flower spikes,commonly known as“nine bends”,which has high economic value.However,the“nine bends”characteristic gradually disappears with age;consequently,the economic value of the Tan sheep decreases.Age-related changes in DNA methylation have been reported and may be responsible for age-induced changes in gene expression.Until now,no genome-wide surveys have been conducted to identify potential DNA methylation sites involved in different sheep growth stages.In this study we investigated the dynamic changes of genome-wide DNA methylation profiles in Tan sheep using DNA from skin and deep whole-genome bisulfite sequencing,and compared the DNA methylation levels at three different growth stages:1,24,and 48 months old(mon1,mon24,and mon48,respectively).Results:In this study,11 skin samples from three growth stages(four for mon1,four for mon24,and three for mon48)were used for DNA methylation analysis and gene expression profiling.There were 52,288 and 236 differentially methylated genes(DMGs)identified between mon1 and mon24,mon1 and mon48,and mon24 and mon48,respectively.Of the differentially methylated regions,1.11%,7.61%,and 7.65% were in the promoter in mon1 vs.mon24,mon24 vs.mon48,and mon1 vs.mon48,respectively.DMGs were enriched in the MAPK and WNT signaling pathways,which are related to age growth and hair follicle morphogenesis processes.There were 51 DMGs associated with age growth and curly fleece formation.Four DMGs between mon1 and mon48(KRT71,CD44,ROR2 and ZDHHC13)were further validated by bisulfite sequencing.Conclusions:This study revealed dynamic changes in the genomic methylation profiles of mon1,mon24,and mon48 sheep,and the percentages of methylated cytosines were 3.38%,2.85% and 4.17%,respectively.Of the DMGs,KRT71 and CD44 were highly methylated in mon1,and ROR2 and ZDHHC13 were highly methylated in mon48.These findings provide foundational information that may be used to develop strategies for potentially retaining the lamb fur and thus improving the economic value of Tan sheep.

Developmental Changes of GH Gene in Tan Sheep and Correlation Analysis with Slaughter Traits

[ Objective] The paper was to study the differential expression of growth hormone （GH） gene in different ages and different tissues of Tan sheep. [ Method] Using real-time fluorescent quantitative PCR （RT-PCR） assay, specific primers were designed according to the GH gene sequence published on Gen- Bank. With the total RNA of Tan sheep tissue as the template, the expression levels of GH gene in pectoral muscle, leg muscle and longissimus dorsi muscle at dif- ferent ages were analyzed. [ Result] In three types of muscle tissues, the expression patterns of GH gene in different tissues at the same month of age were pectoral muscle 〉 leg muscle 〉 longissimus dorsi muscle. The expression levels of GH gene were positively correlated with live weight before slaughter, carcass weight and net meat weight, and had significant positive correlation only with net meat weight （ P 〈 0.05 ）. [ Conclusion ] The content of GH gene in different tissues gradually increased with the increasing months of age, and there were extremely significant differences in expression level of GH gene among different months of age or different sites （P〈0. 01）.

Polymorphism of OB Gene and Its Relationship with Growth Traits in Tan Sheep

In order to promote marker-assisted selection of Tan sheep, the polymorphism of OB gene in zan sheep was detected by sequencing and PCR-RFLP tech- niques. The correlation between polymorphism of OB gene and weight of Tan sheep was analyzed. The results indicated that there was a CT point mutation at locus 253 in exon 3 region of OB gene, but no mutant homozygote was found. The results of chi-square test showed that the frequency of BB and AB genotypes was con- sistent with Hardy-Weinberg equilibrium. AB genotype was the dominant genotype controlling weight of Tan sheep with the genotype frequency of 0.05. According to the results of correlation analysis, the mutation exerted significant effects on weight of Tan sheep at 3-month-old and 6-month-old （P 〈 0.05）. Moreover, interac- tions between ~enotvoe and ~ender had no sianificant effect on weight of Tan sheen （ P 〉 0.05 ）.

SNP Genotyping of Myostatin (MSTN) Gene through Taq Man Probe Assay and Association Study between MSTN Genotypes and Growth Traits of Tan Sheep

To establish a rapid, accurate and economical real-time PCR assay system based on TaqMan probe technology for the detection of genetic variations of single nucleotide polymorphisms （SNPs） in myestatin （MSTN） gene, a pair of TaqMan probes were designed on the polymorphism loci of MSTN gene and used in PCR reaction system for SNP genotyping. Meanwhile, an association study was performed between MSTN genotypes and growth traits of Tan sheep, including birth weight, weaning weight, 3-month weight, and 6-month weight. The results showed that rs417816017 locus of MSTN gene in Tan sheep had two genotypes ： YY and XY. The individuals with genotype XY had a growth advantage over the ones with genotype YY. The results indicate that TaqMan probe-based real-time PCR assay can be used to detect the genotype of MSTN gene, which will provide candidate genes for breeding of Tan sheep.

Countermeasures and Recommendations for High Quality Development of Tan Sheep Industry in Ningxia

Tan sheep industry is a characteristic and superior agricultural industry in Ningxia,and it is also an enriching industry that helps in poverty alleviation and increases farmers’income.The local government has been committing to building the"Yanchi Tan Sheep"brand and has made remarkable achievements.In order to explore the resource efficiency and quality of Tan sheep industry,the current situations and advantages of brand development,this paper made an empirical research through field investigations.The research was mainly concentrated on breeding scale,industrial integration,brand building,breeding benefits,and policy support.After a comprehensive evaluation,it came up with the recommendation that the Tan sheep industry should take the high-quality,high-end and green development road.

Expression of Hoxc13 Gene in Skin Tissue of Tan Sheep at Different Stages

[Objectives]The expression differences of Hoxc13 gene in the skin tissue of Tan sheep at different stages were analyzed. [Methods] qRT-PCR was used to analyze the mRNA expression level of Hoxc13 gene in the skin tissue of Tan sheep at different stages(35, 55 d), and the relative quantitative method was used to analyze the expression level. [Results] The content of Hoxc13 gene in the skin tissue of Tan sheep at 55 d was significantly higher than that in the skin tissue at 35 d. Hoxc13 gene was differentially expressed in the skin tissue of Tan sheep lambs at different stages(35, 55 d). [Conclusions] This study provides basic data for further exploration of the growth and development mechanism of Tan sheep.

Effect of Wolfberry Polysaccharide Additive Premixed Feed on Fattening Effect of Tan Sheep

[Objectives]This study was conducted to explore and rationally use the effective value and nutritional components of wolfberry polysaccharide to provide a basis for the use of new feed resources.[Methods]The premixed feed prepared with the wolfberry polysaccharide additive was applied to fattening Tan sheep in Ningxia,and the effect of the wolfberry polysaccharide additive on the fattening Tan sheep was analyzed,so as to provide technical support for the rational utilization of the wolfberry polysaccharide resources.The study adopted a single-factor comparison method,selecting 84 tan sheep in the fattening period with conform gender,feeding management,weight,age,and physiological status,which were divided into 2 test groups and 1 control group,28 in each group.[Results]Within 90 d,the weight gain of the group with 0.1%wolfberry polysaccharide additive increased by 6.98%,showing a significant difference(P<0.05),and the feed conversion ratio decreased by 9.66%,which was significant as well(P<0.05).The gross profit of this group was 125.76 yuan,which was higher than the control group by 57.52 yuan.The group with 0.2%wolfberry polysaccharide additive increased the weight gain by 8.63%,which was significant(P<0.05),and the feed conversion ratio decreased by 10.80%,which was significant as well(P<0.05).The gross profit of group 2 was 125.76 yuan,which was higher than the control group by 63.41 yuan.There were no significant differences in weight gain,feed conversion ratio and gross profit between the groups with 0.1%and 0.2%of wolfberry polysaccharide additive.The results showed that the use of wolfberry polysaccharide additive premixed feed has a positive effect on increasing the weight gain of fattening Tan sheep,reducing the feed conversion ratio,and improving the feed conversion efficiency.The addition of 0.1%and 0.2%of the wolfberry polysaccharide additive premixed feed had no significant differences.[Conclusions]The wolfberry polysaccharide additive has a positive effect on increasing the weight gain of fattening Tan sheep,reducing the feed conversion ratio,and improving the feed conversion efficiency.

Volatile Flavor Composition in Mutton of Ningxia Tan Sheep

[Objective] To analyze the volatile flavor compounds in mutton of different parts of Ningxia Tan sheep. [Method] The volatile flavor compounds in mutton were identified and quantified using the solid phase micro-extraction （SPME） method combined with GC-MS analysis. [ Re- sult ] The proportion of aldehydes in volatile compounds was the highest in the mutton of most parts of Ningxia Tan sheep, but no 4-methyl acid and 4-methyl nonyl acid was detected in the mutton. [ Conclusion]Aldehydes may be important for the volatile flavor of mutton of Ningxia Tan sheep. The study provides a basis for better development and use of Ningxia Tan sheep.

Polymorphism of Follicle-Stimulating Hormone Receptor Gene in Ningxia Tan Sheep

[ Objective] To study the polymorphism of follicle-stimulating hormone receptor （FSHR） gene in Ningxia Tan sheep and thus to provide a theoretical basis for breeding. I Methodl Genotypes of 111 healthy Ningxia Tan sheep were examined by polymerase chain reaction and restriction fragment length polymorphism （PCR-RFLP）. [ResultS] A 306-bp fragment was amplified. The PCR products digested with restriction enzyme Alu I showed polymorphism with three genotypes, L e., GG, CG and CC. The genotypic frequencies of GG, CG and CC were 0.135 1 （ 15 individuals）, 0.666 7 （74 individuals） and 0.198 2 （22 individuals）, respectively. The allele frequencies of G and C were 0.468 5 and 0.531 5, respectively.[ Conclusion] FSHR aene is Dolvmomhic in Ninaxia Tan Sheeo.

Comparison of Growth and Development Traits among Crossbreds of Dorset and Local Sheep Varieties in Gansu Province

In order to improve the meat production performance of local sheep varieties in Gansu Province, Dorset was introduced to crossbreed with the local sheep varieties, including Tan sheep, Small Tail Han sheep and Mongolia sheep. The offspring of different crossbreeding combinations were sampled randomly at different growth stages, and their growth and development traits were measured so as to screen out the best crossbreeding mode. The results showed that under the same crossbreeding mode, the growth rate of F3 was higher than that of F2, and of F2 was higher than that of F1. Among the F3 population, the growth rates of Dorset ×Han and Dorset × Mongolia hybrids were higher. Compared with those of Dorset ×Tan F3 hybrids, the body weights of male and female Dorset × Han and Dorset ×Mongolia F3 hybrids were increased by 5.59%, 4.40%, 5.93% and 3.76%, respectively. Among the F2 population, the growth rates of Dorset × Han and Dorset ×Mongolia hybrids were also higher. The body weights of male and female Dorset ×Han and Dorset × Mongolia F2 hybrids were higher than those of Dorset × Tan ×Han F2 hybrids by 5.99%, 3.67%, 9.80% and 5.00%, respectively. In the F1 population, the growth rates of Dorset × Han and Dorset × Mongolia hybrids were higher.Compared with those of Tan × Han F1 hybrids, the body weights of male and female Dorset × Han and Dorset × Mongolia F1 hybrids were increased by 11.32%,5.22%, 7.60% and 7.20%, respectively. Therefore, in the feeding area of Small Tai Han sheep, Mongolia sheep and Tan sheep, Dorset was the best sire for producing hybrid lambs. The economic benefit of crossbred offspring was obvious.

Improving the Local Sheep in Gansu via Crossing with Introduced Sheep Breeds Dorset and Borderdale

In order to improve the meat performance of local sheep in Gansu Province,Dorset and Borderdale were introduced to crossbreed with local sheep which were Tan sheep,Small-tail Han sheep and Mongolia sheep. The offspring under different crossbreeding combinations were sampled randomly at the different growing stage to measure their growth traits so as to select optimize the crossbreeding mode. The results indicated that,for the same crossbreeding mode,the growth rate of progeny was in order F3】 F2】 F1; for the F3 progeny,the combinations Dorset- Borderdale- Small tail Han sheep and Dorset- Borderdale- Mongolia sheep gave a higher growth rate,with a body weight of 1. 57%,3. 17%,8. 23%,1. 15% higher in male and female individuals than the counterparts of Dorset and Tan sheep and Small tail Han sheep; for the F2 progeny,the combinations Dorset- Borderdale- Small tail Han sheep and Dorset- Borderdale- Mongolia sheep also gave a higher growth rate,with a body weight of 2. 15%,4. 53%,9. 21% and 2. 75% higher in male and female individuals than the counterparts of Dorset and Tan sheep and Small tail Han sheep; for the F1 progeny,the combination Borderdale and Small tail Han sheep assumed a higher growth rate,with a body weight of 3. 23%,6. 07%,7. 42% and 8. 66% higher in male and female individuals than the counterparts of Borderdale- Mongolia sheep and Tan sheep- Small tail Han sheep,respectively. Therefore,in the Small-tail Han sheep and Mongolia sheep producing regions,the F2 or F3progeny bred by using Dorset or Borderdale sheep as male parent to cross with local breeds,or the hybrid lambs of Small-tail Han sheep and Borderdale sheep as highly qualified commodity,would produce significant economic benefit. Moreover,the novel breeds obtained by crossing were the valuable genetic resource for breeding meat sheep.