At the beginning of the 16th century,Paracelsus coined the maxim:“the dose makes the poison”.This principle can be applied to all living organisms,including organs and cells.The brain and its glial and neuronal cell...At the beginning of the 16th century,Paracelsus coined the maxim:“the dose makes the poison”.This principle can be applied to all living organisms,including organs and cells.The brain and its glial and neuronal cells are no exception.Even small compounds that are essential for the life of brain cells can become truly toxic when overdosed.展开更多
Background The importance of sheep breeding in the Mediterranean part of the eastern Adriatic has a long tradition since its arrival during the Neolithic migrations.Sheep production system is extensive and generally c...Background The importance of sheep breeding in the Mediterranean part of the eastern Adriatic has a long tradition since its arrival during the Neolithic migrations.Sheep production system is extensive and generally carried out in traditional systems without intensive systematic breeding programmes for high uniform trait production(carcass,wool and milk yield).Therefore,eight indigenous Croatian sheep breeds from eastern Adriatic treated here as metapopulation(EAS),are generally considered as multipurpose breeds(milk,meat and wool),not specialised for a particular type of production,but known for their robustness and resistance to certain environmental conditions.Our objective was to identify genomic regions and genes that exhibit patterns of positive selection signatures,decipher their biological and productive functionality,and provide a"genomic"characterization of EAS adaptation and determine its production type.Results We identified positive selection signatures in EAS using several methods based on reduced local variation,linkage disequilibrium and site frequency spectrum(eROHi,iHS,nSL and CLR).Our analyses identified numerous genomic regions and genes(e.g.,desmosomal cadherin and desmoglein gene families)associated with environmental adaptation and economically important traits.Most candidate genes were related to meat/production and health/immune response traits,while some of the candidate genes discovered were important for domestication and evolutionary processes(e.g.,HOXa gene family and FSIP2).These results were also confirmed by GO and QTL enrichment analysis.Conclusions Our results contribute to a better understanding of the unique adaptive genetic architecture of EAS and define its productive type,ultimately providing a new opportunity for future breeding programmes.At the same time,the numerous genes identified will improve our understanding of ruminant(sheep)robustness and resistance in the harsh and specific Mediterranean environment.展开更多
Embryo cryopreservation is an important part of embryo biotechnology.It plays a key role in the development of quality animal husbandry.However,the effect of in-vivo(in-vitro)embryo freezing of cattle and sheep in Chi...Embryo cryopreservation is an important part of embryo biotechnology.It plays a key role in the development of quality animal husbandry.However,the effect of in-vivo(in-vitro)embryo freezing of cattle and sheep in China has not reached an expected level,which limits the commercial application of embryo transfer technology.The technical bottleneck that needs to be broken through is that embryos are subject to mechanical damage and protective agent toxicity during freezing and thawing.This paper summarized the successful application of plant antifreeze protein in vitrification preservation of pig embryos,which makes this technology widely used in animal husbandry production,and is of great significance in accelerating the large-scale breeding of improved livestock and saving endangered animals.展开更多
Medical research on neurologic ailments requires representative animal models to validate treatments before they are translated to human clinical trials.Rodents are the predominant animal model used in neurological re...Medical research on neurologic ailments requires representative animal models to validate treatments before they are translated to human clinical trials.Rodents are the predominant animal model used in neurological research despite limited anatomic and physiologic similarities to humans.As a result,functional testing designed to assess locomotor recovery after neurologic impairment is well established in rodent models.Comparatively,large r,more clinically relevant models have not been as well studied.To achieve similar locomotor testing standardization in larger animals,the models must be accessible to a wide array of researchers.Non-human primates are the most relevant animal model fo r translational research,however ethical and financial barriers limit their accessibility.This review focuses on swine,sheep,and goats as large animal alternatives for transitional studies between rodents and non-human primates.The objective of this review is to compare motor testing and data collection methods used in swine,sheep,and goats to encourage testing standardization in these larger animal models.The PubMed database was analyzed by searching combinations of swine,sheep,and goats,neurologic injuries,and functional assessments.Findings were categorized by animal model,data collection method,and assessment design.Swine and sheep were used in the majority of the studies,while only two studies were found using goats.The functional assessments included open pen analysis,treadmill walking,and guided free walking.Data collection methods included subjective behavioral rating scales and objective tools such as pressure-sensitive mats and image-based analysis software.Overall,swine and sheep were well-suited for a variety of assessment designs,with treadmill walking and guided free walking offering the most consistency across multiple trials.Data collection methods varied,but image-based gait analysis software provided the most robust analysis.Future studies should be conducted to standardize functional testing methods after neurologic impairment in large animals.展开更多
Background Pan-genomics is a recently emerging strategy that can be utilized to provide a more comprehensive characterization of genetic variation.Joint calling is routinely used to combine identified variants across ...Background Pan-genomics is a recently emerging strategy that can be utilized to provide a more comprehensive characterization of genetic variation.Joint calling is routinely used to combine identified variants across multiple related samples.However,the improvement of variants identification using the mutual support information from mul-tiple samples remains quite limited for population-scale genotyping.Results In this study,we developed a computational framework for joint calling genetic variants from 5,061 sheep by incorporating the sequencing error and optimizing mutual support information from multiple samples’data.The variants were accurately identified from multiple samples by using four steps:(1)Probabilities of variants from two widely used algorithms,GATK and Freebayes,were calculated by Poisson model incorporating base sequencing error potential;(2)The variants with high mapping quality or consistently identified from at least two samples by GATK and Freebayes were used to construct the raw high-confidence identification(rHID)variants database;(3)The high confidence variants identified in single sample were ordered by probability value and controlled by false discovery rate(FDR)using rHID database;(4)To avoid the elimination of potentially true variants from rHID database,the vari-ants that failed FDR were reexamined to rescued potential true variants and ensured high accurate identification variants.The results indicated that the percent of concordant SNPs and Indels from Freebayes and GATK after our new method were significantly improved 12%-32%compared with raw variants and advantageously found low frequency variants of individual sheep involved several traits including nipples number(GPC5),scrapie pathology(PAPSS2),sea-sonal reproduction and litter size(GRM1),coat color(RAB27A),and lentivirus susceptibility(TMEM154).Conclusion The new method used the computational strategy to reduce the number of false positives,and simulta-neously improve the identification of genetic variants.This strategy did not incur any extra cost by using any addi-tional samples or sequencing data information and advantageously identified rare variants which can be important for practical applications of animal breeding.展开更多
Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embr...Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.展开更多
In mammals,microRNAs(miRNAs)play key roles in multiple biological processes by regulating the expression of target genes.Studies have found that the levels of miR-370-5p expression differ significantly in the skins of...In mammals,microRNAs(miRNAs)play key roles in multiple biological processes by regulating the expression of target genes.Studies have found that the levels of miR-370-5p expression differ significantly in the skins of sheep with different hair colors;however,its function remains unclear.In this study,we investigated the roles of miR-370-5p in sheep melanocytes and found that the overexpression of miR-370-5p significantly inhibited cell proliferation(P<0.01),tyrosinase activity(P=0.001)and significantly reduced(P<0.001)melanin production.Functional prediction revealed that the 3′-untranslated region(UTR)of MAP3K8 has a putative miR-370-5p binding site,and the interaction between these two molecules was confirmed using luciferase reporter assays.In situ hybridization assays revealed that MAP3K8 is expressed in the cytoplasm of melanocytes.The results of quantitative RT-PCR and Western blotting analyses revealed that overexpression of miR-370-5p in melanocytes significantly inhibits(P<0.01)MAP3K8 expression via direct targeting of its 3′UTR.Inhibition of MAP3K8 expression by siRNA-MAP3K8 transfection induced a significant inhibition(P<0.01)of melanocyte proliferation and significant reduction(P<0.001)in melanin production,which is consistent with our observations for miR-370-5p.Target gene rescue experiments indicated that the expression of MAP3K8 in melanocytes co-transfected with miR-370-5p and MAP3K8-cDNA(containing sites for the targeted binding to miR-370-5p)was significantly rescued(P≤0.001),which subsequently promoted significant increases in cell proliferation(P<0.001)and melanin production(P<0.01).Collectively,these findings indicate that miR-370-5p plays a functional role in inhibiting sheep melanocyte proliferation and melanogenesis by downregulating the expression of MAP3K8.展开更多
Animal genetic resources are playing a vital role in livestock production and are essential to food security. The present study aims to contribute to a better understanding genetic local sheep breeds and to elucidate ...Animal genetic resources are playing a vital role in livestock production and are essential to food security. The present study aims to contribute to a better understanding genetic local sheep breeds and to elucidate the phylogenetic relationships through the evolution of the SRY gene in four different lineages of Ladoum sheep raised in Senegal. After a brief analysis of genetic diversity, the phylogenetic relationships and molecular dating were inferred through haplotype networks and four phylogenetic reconstruction methods. The different haplotype networks are constructed with NETWORK ver. 5.0.0.0 using the Median-Joining method. Phylogenetic trees were reconstructed using neighbor-joining, maximum parsimony, maximum likelihood and Bayesian inference. The robustness of the nodes in phylogenetic trees of the three first methods was assessed by 1000 bootstraps. For Bayesian inference, the posterior probability distribution of the trees was estimated by 4 MCMC chains. 5,000,000 generations were performed for each of the chains by sampling the different parameters every 1000 generations. Results show a low polymorphism. Haplotypic diversity is much higher than the average nucleotide divergence between all pairs of haplotypes. The majority and central haplotype indicates a close relationship between “Batling” and “Tyson” individuals. “Birahim” lineage is very distinct from the rest. Phylogenetic trees confirm two genetically separate clades between “Birahim” and the other lineages. The period of divergence between “Birahim” lineage versus the common ancestor of the other three lineages was 2504 years ago. The polyphyly revealed in “Birahim” lindicates that this lineage does not contain the common ancestor of all individuals who compose it. It could therefore be derived from two or more sheep breeds with a common ancestor, Ovis aries. The monophyletic clade appears to be a group including a common ancestor and all of its genetic descendants. This group, bringing together the other three lineages, is in the process of being structured into sub-lineages. This study is the first to show that there are only two genetic lines within ladoum sheep in Senegal.展开更多
[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 Ta...[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.展开更多
The study was conducted in the Awbarre district of the Fafen zone of the Somali regional state of Ethiopia. The objective of the study was to assess the breeding practices and reproductive performance of Black-head So...The study was conducted in the Awbarre district of the Fafen zone of the Somali regional state of Ethiopia. The objective of the study was to assess the breeding practices and reproductive performance of Black-head Somali sheep under a traditional management system. Purposive and simple random sampling techniques were used to select targeted kebeles and households, respectively. A total of 120 households were selected from four kebeles, each of 30 households, based on the production system and sheep population. Semi-structured questionnaires, group discussions, key informants interviews and field observations were used to generate the required data. The primary purpose of keeping sheep was for income generation, followed by saving as a future asset. The majority (89.2%) of the respondents separated male and female animals during herding. The selection criteria for breeding rams were appearance, growth, pedigree, and color while for breeding ewes were appearance, adaptability, pedigree, color, and lamb growth. The overall weaning age of Black-head Somali sheep in the study area was 3.7 months for both males & females. The castration of male sheep was common for the purpose of fattening, fattening with breeding control and breeding control as well. The castration is mainly performed during the summer and autumn and the methods of castration were both traditional and modern methods, the traditional castration method being the most important one in pastoral areas. The age of sexual maturity was 7.64 months for rams and 8.97 months for ewe’s male and female lambs in the pastoral area and 8.42 & 8.38 for rams & ewes in agro-pastoral and overall lambing interval was 11 months. On average, the ewe of Black-head Somali sheep in pastoral & agro-pastoral could produce 9.49 & 9.57 lambs, respectively in their lifetime. As the pastoralists and agro-pastoralists indicated the source of the breeding ram was their own, so the exchange of breeding ram is recommended to minimize the risk of inbreeding and further studies of on-farm performance investigation would be necessary to be carried out so as to understand the uniqueness of the breed better.展开更多
The present study was conducted to explore ways of JIVET research on Texel sheep,and to provide technical support for the improvement of the JIVET technology system for research on Texel sheep.Superovulation was condu...The present study was conducted to explore ways of JIVET research on Texel sheep,and to provide technical support for the improvement of the JIVET technology system for research on Texel sheep.Superovulation was conducted on 4-8-week old Texel sheep,and in-vivo ovum collection,in-vitro oocyte maturation and fertilization,and fertilized-embryo culture and transplantation were adopted to perform JIVET research on Texel sheep.The results showed that 296 available oocytes were obtained at an average of 74 oocytes per sheep;205 fertilized eggs were obtained at the 2-4 cell stage at an average of 51.25 eggs per sheep and a cleavage rate of 69.26%;and 66 of the fertilized eggs were transplanted to 11 receptor sheep.Six sheep were conceived at a conception rate of 54.55%,and 11 lambs were born.These results indicated that the present test could be used as a method for the study of JIVET technique on Texel sheep.展开更多
This study was conducted to investigate the in-vitro production technology of embryos from young Dorper sheep, so as to provide technical support for the utilization of ovarian follicles in young Dorper sheep. Tests w...This study was conducted to investigate the in-vitro production technology of embryos from young Dorper sheep, so as to provide technical support for the utilization of ovarian follicles in young Dorper sheep. Tests were conducted from the induction of Dorper sheep of 4 to 8 weeks old using follicle stimulating hormone(FSH) and pregnant horse serum(PMSG), collection of oocytes, in-vitro oocyte maturation-fertilization-zygote cultivation and 2-4-cell-stage fertilized ovum transfer. The results showed that 585 oocytes were collected from eight Dorper sheep at the age of 4 and 8 weeks, with an average of 73.13 oocytes/sheep. 346 of the 2-4-cell-stage fertilized eggs were obtained, whose cleavage rate was 59.15%. 77 in-vitro fertilized eggs at 2-4-cell stage were transplanted into 17 recipient sheep, seven of which were pregnant and gave birth to 13 "test-tube sheep" with a conception rate of 41.18%. It is indicated that the hormone induction technique, in-vitro oocyte maturation-fertilization-zygote cultivation technique and 2-4-cell-stage fertilized ovum transfer technique used in this study can serve as effective techniques for the in-vitro production of embryos from Dorper sheep of 4-8 weeks old.展开更多
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 c...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.展开更多
The CRISPR/Cas9 system has been extensively used to engineer genetic loci for the generation of knockouts, insertions, and point mutations in animal models. However, many mutations that have been reported in animals a...The CRISPR/Cas9 system has been extensively used to engineer genetic loci for the generation of knockouts, insertions, and point mutations in animal models. However, many mutations that have been reported in animals are small insertions or deletions. This study used the CRISPR/Cas9 system to induce large DNA fragment deletions in MSTN via three guide RNAs in sheep. This successfully achieved the precise gene editing of the ovine MSTN gene by injecting both Cas9 m RNA and sg RNAs into embryos at the one-cell stage. Of 10 edited animals, 3 animals(30%) exhibited large genomic fragment deletions(~5 kb). Furthermore, the body weights of these 3 animals were significantly different(P0<0.0001, P15=0.001, P30=0.005, P60=0.027) between lambs with large deletions and wildtype lambs. In addition, the edited lambs were also significantly different(P0<0.0001, P15<0.0001, P30=0.002, P60=0.011) compared with wildtype. These results suggest that the generated MSTN knockout sheep is a reliable and effective animal model for further study. Furthermore, this method is time-and labor-saving, and efficient for the creation of animal models for agriculture, biology, and medicine.展开更多
Background: The objectives of this study were to characterize changes in the relative m RNA expression of candidate genes and proteins involved in cell cycle regulation, cell proliferation and apoptosis in the ruminal...Background: The objectives of this study were to characterize changes in the relative m RNA expression of candidate genes and proteins involved in cell cycle regulation, cell proliferation and apoptosis in the ruminal epithelium(RE) of sheep during high-grain(HG) diet adaptation.Results: Twenty sheep were assigned to four groups with five animals each. These animals were assigned to different periods of HG diet(containing 40% forage and 60% concentrate mix) feeding. The HG groups received an HG diet for7(G7, n = 5), 14(G14, n = 5) and 28 d(G28, n = 5), respectively. In contrast, the control group(CON, n = 5) was fed the forage-based diet for 28 d. The results showed that HG feeding linearly decreased(P < 0.001) the ruminal p H, and increased the concentrations of ruminal total volatile fatty acid(linear, P = 0.001), butyrate(linear, P < 0.001), valerate(quadratic P = 0.029) and the level of IGF-1(quadratic, P = 0.043) in plasma. The length(quadratic, P = 0.004), width(cubic, P = 0.015) and surface of the ruminal papillae(linear, P = 0.003) were all enlarged after 14 d of HG diet feeding.HG feeding cubically increased the number of cell layers forming the stratum corneum(SC, P < 0.001) and the thickness of the SC(P < 0.001) and stratum basale(P < 0.001). The proportion of basal layer cells in the RE decreased(linear, P < 0.001) in the G0/G1-phase, but it increased linearly(P = 0.006) in the S-phase and cubically(P = 0.004) in the G2/M-phases. The proportion of apoptosis cells in G7, G14 and G28 was reduced compared to the CON(quadratic, P <0.001). HG diet feeding linearly decreased the m RNA expression of Cyclin E1(P = 0.021) and CDK-2(P = 0.001) and(P = 0.027) the protein expression of Cyclin E1. Feeding an HG diet linearly increased the m RNA expression of genes IGFBP-2(P = 0.034) and IGFBP 5(P < 0.009), while linearly decreasing(P < 0.001) the IGFBP 3 expression. The expression of cell apoptosis gene Caspase 8 decreased(quadratic, P = 0.012), while Bad m RNA expression tended to decrease(cubic, P = 0.053) after HG feeding.Conclusions: These results demonstrated sequential changes in rumen papillae size, cell cycle regulation and the genes involved in proliferation and apoptosis as time elapsed in feeding a high-grain diet to sheep.展开更多
Heat stress significantly impairs reproduction of sheep,and under current climatic conditions is a significant risk to the efficiency of the meat and wool production,with the impact increasing as global temperatures r...Heat stress significantly impairs reproduction of sheep,and under current climatic conditions is a significant risk to the efficiency of the meat and wool production,with the impact increasing as global temperatures rise.Evidence from field studies and studies conducted using environmental chambers demonstrate the effects of hot temperatures(≥32℃)on components of ewe fertility(oestrus,fertilisation,embryo survival and lambing)are most destructive when experienced from 5 d before until 5 d after oestrus.Temperature controlled studies also demonstrate that ram fertility,as measured by rates of fertilisation and embryo survival,is reduced when mating occurs during the period 14 to 50 d post-heating.However,the contribution of the ram to heat induced reductions in flock fertility is difficult to determine accurately.Based primarily on temperature controlled studies,it is clear that sustained exposure to high temperatures(≥32℃)during pregnancy reduces lamb birthweight and will,therefore,decrease lamb survival under field conditions.It is concluded that both ewe and ram reproduction is affected by relatively modest levels of heat stress(≥32℃)and this is a concern given that a significant proportion of the global sheep population experiences heat stress of this magnitude around mating and during pregnancy.Despite this,strategies to limit the impacts of the climate on the homeothermy,behaviour,resource use and reproduction of extensively grazed sheep are limited,and there is an urgency to improve knowledge and to develop husbandry practices to limit these impacts.展开更多
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 offsprin...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.展开更多
Background: The widely observed RNA-DNA differences(RDDs) have been found to be due to nucleotide alteration by RNA editing. Canonical RNA editing(i.e., A-to-I and C-to-U editing) mediated by the adenosine deaminases ...Background: The widely observed RNA-DNA differences(RDDs) have been found to be due to nucleotide alteration by RNA editing. Canonical RNA editing(i.e., A-to-I and C-to-U editing) mediated by the adenosine deaminases acting on RNA(ADAR) family and apolipoprotein B mRNA editing catalytic polypeptide-like(APOBEC)family during the transcriptional process is considered common and essential for the development of an individual.To date, an increasing number of RNA editing sites have been reported in human, rodents, and some farm animals;however, genome-wide detection of RNA editing events in sheep has not been reported. The aim of this study was to identify RNA editing events in sheep by comparing the RNA-seq and DNA-seq data from three biological replicates of the kidney and spleen tissues.Results: A total of 607 and 994 common edited sites within the three biological replicates were identified in the ovine kidney and spleen, respectively. Many of the RDDs were specific to an individual. The RNA editing-related genes identified in the present study might be evolved for specific biological functions in sheep, such as structural constituent of the cytoskeleton and microtubule-based processes. Furthermore, the edited sites found in the ovine BLCAP and NEIL1 genes are in line with those in previous reports on the porcine and human homologs, suggesting the existence of evolutionarily conserved RNA editing sites and they may play an important role in the structure and function of genes.Conclusions: Our study is the first to investigate RNA editing events in sheep. We screened out 607 and 994 RNA editing sites in three biological replicates of the ovine kidney and spleen and annotated 164 and 247 genes in the kidney and spleen, respectively. The gene function and conservation analysis of these RNA editing-related genes suggest that RNA editing is associated with important gene function in sheep. The putative functionally important RNA editing sites reported in the present study will help future studies on the relationship between these edited sites and the genetic traits in sheep.展开更多
Background: Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell ac...Background: Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. Consequently, transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis. In this study, we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons(breeding and non-breeding season) on superovulation and the imported exogenous gene on growth.Results: In total of 43 donor ewes and 166 recipient ewes in breeding season, 37 donor ewes and 144 recipient ewes in non-breeding season were selected for super-ovulation and injected embryo transfer to generate transgenic sheep.Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons(P > 0.05). The positive rate of exogenous TLR4 tested were 21.21 % and 22.58 % in breeding and non-breeding season by Southern blot. The expression level of TLR4 in the transgenic sheep was 1.5 times higher than in the non-transgenic group(P < 0.05). The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.Conclusions: Here we establish an efficient platform for the production of transgenic sheep by the microinjection of pronuclear embryos during the whole year. The over-expression of TLR4 had no adverse effect on the growth of the sheep.展开更多
文摘At the beginning of the 16th century,Paracelsus coined the maxim:“the dose makes the poison”.This principle can be applied to all living organisms,including organs and cells.The brain and its glial and neuronal cells are no exception.Even small compounds that are essential for the life of brain cells can become truly toxic when overdosed.
基金supported by Croatian Science Foundation project IP-2018–01-8708-Application of NGS methods in the assessment of genomic variability in ruminants–“ANAGRAMS”the EU Operational Program Competitiveness and Cohesion 2014–2020 project KK.01.1.1.04.0058—Potential of microencapsulation in cheese productionthe project No.QK1919156 of the Ministry of Agriculture,Czech Republic.
文摘Background The importance of sheep breeding in the Mediterranean part of the eastern Adriatic has a long tradition since its arrival during the Neolithic migrations.Sheep production system is extensive and generally carried out in traditional systems without intensive systematic breeding programmes for high uniform trait production(carcass,wool and milk yield).Therefore,eight indigenous Croatian sheep breeds from eastern Adriatic treated here as metapopulation(EAS),are generally considered as multipurpose breeds(milk,meat and wool),not specialised for a particular type of production,but known for their robustness and resistance to certain environmental conditions.Our objective was to identify genomic regions and genes that exhibit patterns of positive selection signatures,decipher their biological and productive functionality,and provide a"genomic"characterization of EAS adaptation and determine its production type.Results We identified positive selection signatures in EAS using several methods based on reduced local variation,linkage disequilibrium and site frequency spectrum(eROHi,iHS,nSL and CLR).Our analyses identified numerous genomic regions and genes(e.g.,desmosomal cadherin and desmoglein gene families)associated with environmental adaptation and economically important traits.Most candidate genes were related to meat/production and health/immune response traits,while some of the candidate genes discovered were important for domestication and evolutionary processes(e.g.,HOXa gene family and FSIP2).These results were also confirmed by GO and QTL enrichment analysis.Conclusions Our results contribute to a better understanding of the unique adaptive genetic architecture of EAS and define its productive type,ultimately providing a new opportunity for future breeding programmes.At the same time,the numerous genes identified will improve our understanding of ruminant(sheep)robustness and resistance in the harsh and specific Mediterranean environment.
基金Supported by Key Project of Qiqihar Science and Technology Program(ZDTG-202201)Qiqihar Innovation Incentive Project(CNGG-2021015)。
文摘Embryo cryopreservation is an important part of embryo biotechnology.It plays a key role in the development of quality animal husbandry.However,the effect of in-vivo(in-vitro)embryo freezing of cattle and sheep in China has not reached an expected level,which limits the commercial application of embryo transfer technology.The technical bottleneck that needs to be broken through is that embryos are subject to mechanical damage and protective agent toxicity during freezing and thawing.This paper summarized the successful application of plant antifreeze protein in vitrification preservation of pig embryos,which makes this technology widely used in animal husbandry production,and is of great significance in accelerating the large-scale breeding of improved livestock and saving endangered animals.
文摘Medical research on neurologic ailments requires representative animal models to validate treatments before they are translated to human clinical trials.Rodents are the predominant animal model used in neurological research despite limited anatomic and physiologic similarities to humans.As a result,functional testing designed to assess locomotor recovery after neurologic impairment is well established in rodent models.Comparatively,large r,more clinically relevant models have not been as well studied.To achieve similar locomotor testing standardization in larger animals,the models must be accessible to a wide array of researchers.Non-human primates are the most relevant animal model fo r translational research,however ethical and financial barriers limit their accessibility.This review focuses on swine,sheep,and goats as large animal alternatives for transitional studies between rodents and non-human primates.The objective of this review is to compare motor testing and data collection methods used in swine,sheep,and goats to encourage testing standardization in these larger animal models.The PubMed database was analyzed by searching combinations of swine,sheep,and goats,neurologic injuries,and functional assessments.Findings were categorized by animal model,data collection method,and assessment design.Swine and sheep were used in the majority of the studies,while only two studies were found using goats.The functional assessments included open pen analysis,treadmill walking,and guided free walking.Data collection methods included subjective behavioral rating scales and objective tools such as pressure-sensitive mats and image-based analysis software.Overall,swine and sheep were well-suited for a variety of assessment designs,with treadmill walking and guided free walking offering the most consistency across multiple trials.Data collection methods varied,but image-based gait analysis software provided the most robust analysis.Future studies should be conducted to standardize functional testing methods after neurologic impairment in large animals.
基金Superior Farms sheep producersIBEST for their supportfinancial support from the Idaho Global Entrepreneurial Mission
文摘Background Pan-genomics is a recently emerging strategy that can be utilized to provide a more comprehensive characterization of genetic variation.Joint calling is routinely used to combine identified variants across multiple related samples.However,the improvement of variants identification using the mutual support information from mul-tiple samples remains quite limited for population-scale genotyping.Results In this study,we developed a computational framework for joint calling genetic variants from 5,061 sheep by incorporating the sequencing error and optimizing mutual support information from multiple samples’data.The variants were accurately identified from multiple samples by using four steps:(1)Probabilities of variants from two widely used algorithms,GATK and Freebayes,were calculated by Poisson model incorporating base sequencing error potential;(2)The variants with high mapping quality or consistently identified from at least two samples by GATK and Freebayes were used to construct the raw high-confidence identification(rHID)variants database;(3)The high confidence variants identified in single sample were ordered by probability value and controlled by false discovery rate(FDR)using rHID database;(4)To avoid the elimination of potentially true variants from rHID database,the vari-ants that failed FDR were reexamined to rescued potential true variants and ensured high accurate identification variants.The results indicated that the percent of concordant SNPs and Indels from Freebayes and GATK after our new method were significantly improved 12%-32%compared with raw variants and advantageously found low frequency variants of individual sheep involved several traits including nipples number(GPC5),scrapie pathology(PAPSS2),sea-sonal reproduction and litter size(GRM1),coat color(RAB27A),and lentivirus susceptibility(TMEM154).Conclusion The new method used the computational strategy to reduce the number of false positives,and simulta-neously improve the identification of genetic variants.This strategy did not incur any extra cost by using any addi-tional samples or sequencing data information and advantageously identified rare variants which can be important for practical applications of animal breeding.
基金financially supported by the Science and Technology Major Project of the Inner Mongolia Autonomous Region of China(2020ZD0007)the Major Program of the Inner Mongolia Natural Science Foundation,China(2020ZD10)+3 种基金the National Natural Science Foundation of China(32160172)the Natural Science Foundation of Inner Mongolia Autonomous Region(2020BS03003 and 2020BS03022)the National Transgenic Project of China(2016ZX0801000-002 and 2016ZX08010005-001)the Science and Technology Major Project of the Inner Mongolia Autonomous Region of China(zdzx2018065)。
文摘Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.
基金funded by the Natural Science Foundation of Anhui Province,China(2008085QC158)the University Natural Science Research Project of Anhui Province(KJ2019A0165)。
文摘In mammals,microRNAs(miRNAs)play key roles in multiple biological processes by regulating the expression of target genes.Studies have found that the levels of miR-370-5p expression differ significantly in the skins of sheep with different hair colors;however,its function remains unclear.In this study,we investigated the roles of miR-370-5p in sheep melanocytes and found that the overexpression of miR-370-5p significantly inhibited cell proliferation(P<0.01),tyrosinase activity(P=0.001)and significantly reduced(P<0.001)melanin production.Functional prediction revealed that the 3′-untranslated region(UTR)of MAP3K8 has a putative miR-370-5p binding site,and the interaction between these two molecules was confirmed using luciferase reporter assays.In situ hybridization assays revealed that MAP3K8 is expressed in the cytoplasm of melanocytes.The results of quantitative RT-PCR and Western blotting analyses revealed that overexpression of miR-370-5p in melanocytes significantly inhibits(P<0.01)MAP3K8 expression via direct targeting of its 3′UTR.Inhibition of MAP3K8 expression by siRNA-MAP3K8 transfection induced a significant inhibition(P<0.01)of melanocyte proliferation and significant reduction(P<0.001)in melanin production,which is consistent with our observations for miR-370-5p.Target gene rescue experiments indicated that the expression of MAP3K8 in melanocytes co-transfected with miR-370-5p and MAP3K8-cDNA(containing sites for the targeted binding to miR-370-5p)was significantly rescued(P≤0.001),which subsequently promoted significant increases in cell proliferation(P<0.001)and melanin production(P<0.01).Collectively,these findings indicate that miR-370-5p plays a functional role in inhibiting sheep melanocyte proliferation and melanogenesis by downregulating the expression of MAP3K8.
文摘Animal genetic resources are playing a vital role in livestock production and are essential to food security. The present study aims to contribute to a better understanding genetic local sheep breeds and to elucidate the phylogenetic relationships through the evolution of the SRY gene in four different lineages of Ladoum sheep raised in Senegal. After a brief analysis of genetic diversity, the phylogenetic relationships and molecular dating were inferred through haplotype networks and four phylogenetic reconstruction methods. The different haplotype networks are constructed with NETWORK ver. 5.0.0.0 using the Median-Joining method. Phylogenetic trees were reconstructed using neighbor-joining, maximum parsimony, maximum likelihood and Bayesian inference. The robustness of the nodes in phylogenetic trees of the three first methods was assessed by 1000 bootstraps. For Bayesian inference, the posterior probability distribution of the trees was estimated by 4 MCMC chains. 5,000,000 generations were performed for each of the chains by sampling the different parameters every 1000 generations. Results show a low polymorphism. Haplotypic diversity is much higher than the average nucleotide divergence between all pairs of haplotypes. The majority and central haplotype indicates a close relationship between “Batling” and “Tyson” individuals. “Birahim” lineage is very distinct from the rest. Phylogenetic trees confirm two genetically separate clades between “Birahim” and the other lineages. The period of divergence between “Birahim” lineage versus the common ancestor of the other three lineages was 2504 years ago. The polyphyly revealed in “Birahim” lindicates that this lineage does not contain the common ancestor of all individuals who compose it. It could therefore be derived from two or more sheep breeds with a common ancestor, Ovis aries. The monophyletic clade appears to be a group including a common ancestor and all of its genetic descendants. This group, bringing together the other three lineages, is in the process of being structured into sub-lineages. This study is the first to show that there are only two genetic lines within ladoum sheep in Senegal.
基金Supported by Ningxia Agricultural Reclamation Group Science and Technology Innovation ProjectNew Feed Technology Promotion Project of Ningxia Agriculture and Rural DepartmentNingxia Feed Industry Expert Technical Service Group Project。
文摘[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.
文摘The study was conducted in the Awbarre district of the Fafen zone of the Somali regional state of Ethiopia. The objective of the study was to assess the breeding practices and reproductive performance of Black-head Somali sheep under a traditional management system. Purposive and simple random sampling techniques were used to select targeted kebeles and households, respectively. A total of 120 households were selected from four kebeles, each of 30 households, based on the production system and sheep population. Semi-structured questionnaires, group discussions, key informants interviews and field observations were used to generate the required data. The primary purpose of keeping sheep was for income generation, followed by saving as a future asset. The majority (89.2%) of the respondents separated male and female animals during herding. The selection criteria for breeding rams were appearance, growth, pedigree, and color while for breeding ewes were appearance, adaptability, pedigree, color, and lamb growth. The overall weaning age of Black-head Somali sheep in the study area was 3.7 months for both males & females. The castration of male sheep was common for the purpose of fattening, fattening with breeding control and breeding control as well. The castration is mainly performed during the summer and autumn and the methods of castration were both traditional and modern methods, the traditional castration method being the most important one in pastoral areas. The age of sexual maturity was 7.64 months for rams and 8.97 months for ewe’s male and female lambs in the pastoral area and 8.42 & 8.38 for rams & ewes in agro-pastoral and overall lambing interval was 11 months. On average, the ewe of Black-head Somali sheep in pastoral & agro-pastoral could produce 9.49 & 9.57 lambs, respectively in their lifetime. As the pastoralists and agro-pastoralists indicated the source of the breeding ram was their own, so the exchange of breeding ram is recommended to minimize the risk of inbreeding and further studies of on-farm performance investigation would be necessary to be carried out so as to understand the uniqueness of the breed better.
基金the Science and Technology Project of Shanxi Province(20120311024-1)Special Fund for the Construction of National Wool Sheep Industrial Technology Research System(CARS-39-24)+1 种基金Shanxi Provincial Science and Technology Innovation Team Fund(201705D131028-20)Shanxi Financial Fund for Agriculture(NYGX2015-03)。
文摘The present study was conducted to explore ways of JIVET research on Texel sheep,and to provide technical support for the improvement of the JIVET technology system for research on Texel sheep.Superovulation was conducted on 4-8-week old Texel sheep,and in-vivo ovum collection,in-vitro oocyte maturation and fertilization,and fertilized-embryo culture and transplantation were adopted to perform JIVET research on Texel sheep.The results showed that 296 available oocytes were obtained at an average of 74 oocytes per sheep;205 fertilized eggs were obtained at the 2-4 cell stage at an average of 51.25 eggs per sheep and a cleavage rate of 69.26%;and 66 of the fertilized eggs were transplanted to 11 receptor sheep.Six sheep were conceived at a conception rate of 54.55%,and 11 lambs were born.These results indicated that the present test could be used as a method for the study of JIVET technique on Texel sheep.
基金Supported by Earmarked Fund for Construction of National Wool Sheep Industry Technology Research System(CARS-39-24)Program for Science and Technology Development of Shanxi Province(20120311024-1)+1 种基金Fund for Science and Technology Innovation Team in Shanxi Province(201705D131028-20)Shanxi Agricultural Industry Development Technology Leading Fund(2017CYYL-08)
文摘This study was conducted to investigate the in-vitro production technology of embryos from young Dorper sheep, so as to provide technical support for the utilization of ovarian follicles in young Dorper sheep. Tests were conducted from the induction of Dorper sheep of 4 to 8 weeks old using follicle stimulating hormone(FSH) and pregnant horse serum(PMSG), collection of oocytes, in-vitro oocyte maturation-fertilization-zygote cultivation and 2-4-cell-stage fertilized ovum transfer. The results showed that 585 oocytes were collected from eight Dorper sheep at the age of 4 and 8 weeks, with an average of 73.13 oocytes/sheep. 346 of the 2-4-cell-stage fertilized eggs were obtained, whose cleavage rate was 59.15%. 77 in-vitro fertilized eggs at 2-4-cell stage were transplanted into 17 recipient sheep, seven of which were pregnant and gave birth to 13 "test-tube sheep" with a conception rate of 41.18%. It is indicated that the hormone induction technique, in-vitro oocyte maturation-fertilization-zygote cultivation technique and 2-4-cell-stage fertilized ovum transfer technique used in this study can serve as effective techniques for the in-vitro production of embryos from Dorper sheep of 4-8 weeks old.
基金Supported by Special Project for Agricultural Breeding of Ningxia(2018NYYZ0401)Agricultural High Quality Development and Ecological Protection Science and Technology Innovation Demonstration Project of Ningxia(NGSB-2020-12)Scientific and Technological Research Program of Special Project for Agricultural Science and Technology Independent Innovation of Ningxia(NKYZZ-G-19-08)。
文摘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.
基金supported by the grants from the National Natural Science Foundation of China(31572369,31772571,31872332)the earmarked fund for China Agriculture Research System(CARS-39-12)the Tan Sheep Breeding Project of Ningxia,China(NXTS201601)。
文摘The CRISPR/Cas9 system has been extensively used to engineer genetic loci for the generation of knockouts, insertions, and point mutations in animal models. However, many mutations that have been reported in animals are small insertions or deletions. This study used the CRISPR/Cas9 system to induce large DNA fragment deletions in MSTN via three guide RNAs in sheep. This successfully achieved the precise gene editing of the ovine MSTN gene by injecting both Cas9 m RNA and sg RNAs into embryos at the one-cell stage. Of 10 edited animals, 3 animals(30%) exhibited large genomic fragment deletions(~5 kb). Furthermore, the body weights of these 3 animals were significantly different(P0<0.0001, P15=0.001, P30=0.005, P60=0.027) between lambs with large deletions and wildtype lambs. In addition, the edited lambs were also significantly different(P0<0.0001, P15<0.0001, P30=0.002, P60=0.011) compared with wildtype. These results suggest that the generated MSTN knockout sheep is a reliable and effective animal model for further study. Furthermore, this method is time-and labor-saving, and efficient for the creation of animal models for agriculture, biology, and medicine.
基金supported by the National Natural Science Foundation of China (No.31572436)Natural Science Foundation of China (No. 31372339)
文摘Background: The objectives of this study were to characterize changes in the relative m RNA expression of candidate genes and proteins involved in cell cycle regulation, cell proliferation and apoptosis in the ruminal epithelium(RE) of sheep during high-grain(HG) diet adaptation.Results: Twenty sheep were assigned to four groups with five animals each. These animals were assigned to different periods of HG diet(containing 40% forage and 60% concentrate mix) feeding. The HG groups received an HG diet for7(G7, n = 5), 14(G14, n = 5) and 28 d(G28, n = 5), respectively. In contrast, the control group(CON, n = 5) was fed the forage-based diet for 28 d. The results showed that HG feeding linearly decreased(P < 0.001) the ruminal p H, and increased the concentrations of ruminal total volatile fatty acid(linear, P = 0.001), butyrate(linear, P < 0.001), valerate(quadratic P = 0.029) and the level of IGF-1(quadratic, P = 0.043) in plasma. The length(quadratic, P = 0.004), width(cubic, P = 0.015) and surface of the ruminal papillae(linear, P = 0.003) were all enlarged after 14 d of HG diet feeding.HG feeding cubically increased the number of cell layers forming the stratum corneum(SC, P < 0.001) and the thickness of the SC(P < 0.001) and stratum basale(P < 0.001). The proportion of basal layer cells in the RE decreased(linear, P < 0.001) in the G0/G1-phase, but it increased linearly(P = 0.006) in the S-phase and cubically(P = 0.004) in the G2/M-phases. The proportion of apoptosis cells in G7, G14 and G28 was reduced compared to the CON(quadratic, P <0.001). HG diet feeding linearly decreased the m RNA expression of Cyclin E1(P = 0.021) and CDK-2(P = 0.001) and(P = 0.027) the protein expression of Cyclin E1. Feeding an HG diet linearly increased the m RNA expression of genes IGFBP-2(P = 0.034) and IGFBP 5(P < 0.009), while linearly decreasing(P < 0.001) the IGFBP 3 expression. The expression of cell apoptosis gene Caspase 8 decreased(quadratic, P = 0.012), while Bad m RNA expression tended to decrease(cubic, P = 0.053) after HG feeding.Conclusions: These results demonstrated sequential changes in rumen papillae size, cell cycle regulation and the genes involved in proliferation and apoptosis as time elapsed in feeding a high-grain diet to sheep.
基金This review was funded as part of a Meat&Livestock Australia Ltd.tender entitled The effects of heat stress and predicted climate change scenarios on reproductive performance of the Australia sheep flock(Project code:L.LSM.0024).
文摘Heat stress significantly impairs reproduction of sheep,and under current climatic conditions is a significant risk to the efficiency of the meat and wool production,with the impact increasing as global temperatures rise.Evidence from field studies and studies conducted using environmental chambers demonstrate the effects of hot temperatures(≥32℃)on components of ewe fertility(oestrus,fertilisation,embryo survival and lambing)are most destructive when experienced from 5 d before until 5 d after oestrus.Temperature controlled studies also demonstrate that ram fertility,as measured by rates of fertilisation and embryo survival,is reduced when mating occurs during the period 14 to 50 d post-heating.However,the contribution of the ram to heat induced reductions in flock fertility is difficult to determine accurately.Based primarily on temperature controlled studies,it is clear that sustained exposure to high temperatures(≥32℃)during pregnancy reduces lamb birthweight and will,therefore,decrease lamb survival under field conditions.It is concluded that both ewe and ram reproduction is affected by relatively modest levels of heat stress(≥32℃)and this is a concern given that a significant proportion of the global sheep population experiences heat stress of this magnitude around mating and during pregnancy.Despite this,strategies to limit the impacts of the climate on the homeothermy,behaviour,resource use and reproduction of extensively grazed sheep are limited,and there is an urgency to improve knowledge and to develop husbandry practices to limit these impacts.
基金Supported by Science&Technology Pillar Program of Gansu Province(1104NKCA083)
文摘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.
基金funded by the National Nature Science Foundation of China(U1136605)Ningxia Agricultural Breeding Program(NXNYYZ20150103)Program for Changjiang Scholar and Innovation Research Team in University(IRT1191)
文摘Background: The widely observed RNA-DNA differences(RDDs) have been found to be due to nucleotide alteration by RNA editing. Canonical RNA editing(i.e., A-to-I and C-to-U editing) mediated by the adenosine deaminases acting on RNA(ADAR) family and apolipoprotein B mRNA editing catalytic polypeptide-like(APOBEC)family during the transcriptional process is considered common and essential for the development of an individual.To date, an increasing number of RNA editing sites have been reported in human, rodents, and some farm animals;however, genome-wide detection of RNA editing events in sheep has not been reported. The aim of this study was to identify RNA editing events in sheep by comparing the RNA-seq and DNA-seq data from three biological replicates of the kidney and spleen tissues.Results: A total of 607 and 994 common edited sites within the three biological replicates were identified in the ovine kidney and spleen, respectively. Many of the RDDs were specific to an individual. The RNA editing-related genes identified in the present study might be evolved for specific biological functions in sheep, such as structural constituent of the cytoskeleton and microtubule-based processes. Furthermore, the edited sites found in the ovine BLCAP and NEIL1 genes are in line with those in previous reports on the porcine and human homologs, suggesting the existence of evolutionarily conserved RNA editing sites and they may play an important role in the structure and function of genes.Conclusions: Our study is the first to investigate RNA editing events in sheep. We screened out 607 and 994 RNA editing sites in three biological replicates of the ovine kidney and spleen and annotated 164 and 247 genes in the kidney and spleen, respectively. The gene function and conservation analysis of these RNA editing-related genes suggest that RNA editing is associated with important gene function in sheep. The putative functionally important RNA editing sites reported in the present study will help future studies on the relationship between these edited sites and the genetic traits in sheep.
基金supported by grants from National Transgenic Creature Breeding Grand Project (2014ZX08008-005)
文摘Background: Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. Consequently, transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis. In this study, we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons(breeding and non-breeding season) on superovulation and the imported exogenous gene on growth.Results: In total of 43 donor ewes and 166 recipient ewes in breeding season, 37 donor ewes and 144 recipient ewes in non-breeding season were selected for super-ovulation and injected embryo transfer to generate transgenic sheep.Our results indicated the no. of embryos recovered of donors and the rate of pronuclear embryos did not show any significant difference between breeding and non-breeding seasons(P > 0.05). The positive rate of exogenous TLR4 tested were 21.21 % and 22.58 % in breeding and non-breeding season by Southern blot. The expression level of TLR4 in the transgenic sheep was 1.5 times higher than in the non-transgenic group(P < 0.05). The lambs overexpressing TLR4 had similar growth performance with non-transgenic lambs, and the blood physiological parameters of transgenic and non-transgenic were both in the normal range and did not show any difference.Conclusions: Here we establish an efficient platform for the production of transgenic sheep by the microinjection of pronuclear embryos during the whole year. The over-expression of TLR4 had no adverse effect on the growth of the sheep.
