本文对不同饲料利用效率大白猪的肉质性状进行测定,同时利用实时荧光定量PCR检测背最长肌和皮下脂肪中与肉质、脂肪沉积性状相关基因的转录水平,分析FASN和PPARγ基因表达量与肉质性状的相关性,结果表明:不同饲料利用效率大白猪肉品质中pH24h值为 5.63～5.81;L24h值为 48.43～50.02;b24h值为6.90～7.82;48 h滴水损失为 3.62～3.99;肌内脂肪为 1.59～2.21.实时荧光定量PCR测定结果显示,在不同饲料利用效率大白猪背最长肌中,PPARγ和FASN基因的mRNA表达水平分别为差异显著和极显著,二者与肌内脂肪含量呈正相关;肉质性状相关性分析及肉质性状与基因表达量相关性分析中,高饲料利用效率(High Feed Efficiency,HFE)大白猪的肉色L值与pH值呈显著负相关;HFE大白猪背最长肌中FASN基因mRNA表达量与L值呈显著正相关,在低饲料利用效率(Low Feed Efficiency,LFE)大白猪背最长肌中PPARγ基因mRNA表达量与L值呈极显著正相关.

为探究NLR Family Pyrin Domain Containing 6(NLRP6)在广西巴马小型猪的蛋白结构和组织表达水平,本实验以巴马小型猪为材料,通过克隆获取了NLRP6的CDS序列全长,并进行生物信息学和组织表达谱分析,同时采用qRT-PCR分析NLRP6基因在不同组织的表达分布情况.结果显示,NLRP6基因的CDS序列全长为 2640 bp,编码 879 个氨基酸.生物信息学分析表明,NLRP6 蛋白分子式预测为C4294H6958N1234O1272S37,等电点为 8.12,为亲水性碱性蛋白质.经氨基酸序列比对和物种进化树分析发现,广西巴马小型猪NLRP6基因与猪的遗传距离最近,其次是牛,与小鼠的遗传距离最远.qRT-PCR结果表明,NLRP6基因在肾脏组织表达最高,同时在小肠部位具有很高的表达水平,但在心脏和肌肉组织中表达量极低.本研究结果为进一步研究猪NLRP6基因提供了理论参考.

【Objective】 This study was aimed to clone the cysteine dioxygenase type 1（CDO1） gene and conduct bioinformatics analysis, detect the expression of CDO1 gene in different tissues of Large White pigs, and locate the site of CDO1 in mammary gland, so as to provide the basis for further exploring the regulatory role of CDO1 gene in mammary gland development.【Method】 The full-length sequence of CDO1 gene CDS in Large White pigs was amplified by PCR.The CDO1 gene sequence was ligated with pMD18-T vector to transformed into E.coli DH5α competent cells.After PCR identification and sequencing of the amplified culture solution of positive bacteria, the sequence similarity among different species were compared and the phylogenetic tree was constructed.The bioinformatics analysis of CDO1 protein was carried out using online prediction software.The expression of CDO1 gene in different tissues of Large White pigs was detected by Real-time quantitative PCR,the localization of CDO1 protein in the mammary gland of Large White pigs was detected by immunohistochemistry.【Result】 The full-length sequence of CDO1 gene CDS in Large White pigs was 603 bp, encoding 200 amino acids.There was the highest similarity with the nucleotide sequence of CDO1 gene between Large White pigs and Sus scrofa.The phylogenetic tree results showed that Large White pigs and Sus scrofa were first clustered into a group and were closely related to Macaca mulatta.The molecular mass of CDO1 protein was 23.018 ku, the isoelectric point was 5.98,and the instability coefficient was 33.58（＜40）,which was a stable hydrophilic protein and located in the cytoplasm.CDO1 protein had glycosylation and phosphorylation sites.The secondary structure and tertiary structure model of CDO1 protein were consistent.Tissue expression analysis results showed that the expression of CDO1 gene in liver of Large White pigs was the highest, which was significantly higher than that in other tissues（P＜0.05）,it was next expressed in mammary gland, longissimus dorsi muscle and heart, which were significantly higher than that in other tissues except for liver（P＜0.05）,the expression of CDO1 gene in duodenum was the lowest.Immunohistochemical results showed that CDO1 was localized in somatic cells, epithelial cells and adipocytes of terminal bud（TEB） of mammary gland in sows.【Conclusion 】 The total length of CDO1 gene CDS in Large White pigs was 603 bp, encoding 200 amino acids, which was highly expressed in heart, liver and mammary gland.CDO1 protein was a stable hydrophilic protein, which was enriched in somatic cells, epithelial cells and adipocytes of TEB of mammary gland in sows.The results provided a reference for further exploring the effect of CDO1 protein on mammary gland development and lactation function of sows.

为了探究胆汁酸代谢途径关键基因FXR在大白猪中的结构和功能,本研究利用 3 头大白猪十二指肠、空肠、回肠、盲肠、结肠和直肠组织,分别提取RNA,将RNA反转录成cDNA后做混池通过PCR扩增FXR基因的CDS区域,对测序结果进行比对确认,利用多种生物信息学软件对大白猪FXR基因的CDS区域进行了一系列分析.结果显示,FXR基因CDS全长 1461 bp,编码 486 个氨基酸,预测FXR蛋白二级和三级结构,该蛋白的二级结构有 23 个α螺旋、28 个β折叠、30 个T转角、20 个无规卷曲,为亲水性膜外蛋白;并且有多个磷酸化位点;荧光定量PCR结果显示,FXR基因在大白猪回肠中表达量极显著高于其他肠道组织.本实验成功克隆了大白猪FXR基因的CDS区域,为进一步探究FXR基因在猪胆汁酸代谢途径中的调控机制提供了研究基础,也为利用胆汁酸代谢调控挖掘提高猪生长性能的分子标记提供了一定参考.

[目的]获取巴马小型猪ATP结合盒D亚科成员4(ATP binding cassette subfamily D member 4,ABCD4)基因CDS区序列并预测其编码蛋白的结构功能,构建真核表达载体,了解ABCD4基因在巴马小型猪中的组织分布情况,为探究ABC D4基因对巴马小型猪生长发育的作用提供理论和分子基础.[方法]以巴马小型猪皮下脂肪组织cDNA为模板,利用RT-PCR对A BCD4基因CDS区进行扩增、测序,利用生物信息学分析软件对巴马小型猪ABCD4基因CDS区序列与不同物种比对,构建系统进化树,并对ABCD4蛋白的理化性质、跨膜结构等进行预测.将获得的目的基因连接至pEGFP-N1载体并转染到IPEC-J2细胞,检测转染后荧光和基因表达量变化.通过实时荧光定量PCR检测巴马小型猪各组织A BCD4基因表达量.[结果]巴马小型猪ABCD4基因CDS区全长1 818 bp,编码605个氨基酸,与Ensembl上猪参考序列(ENSSSCT00000037529)的相似性为99.7％,存在6处碱基突变和1处碱基插入.相似性比对发现,猪和人、原鸡、倭黑猩猩、马、双峰驼、牛ABCD4序列的相似性分别为87.6％、72.7％、87.6％、91.7％、88.1％和88.6％.系统进化树显示,猪与马亲缘关系最近,与原鸡亲缘关系最远.生物信息学分析显示,ABCD4蛋白分子质量为68.66 ku,理论等电点(pI)为7.13,存在5个跨膜螺旋结构,无信号肽,属于跨膜蛋白;存在58个磷酸化位点和10个N-糖基化位点;二级结构中α-螺旋、β-转角、无规则卷曲、延伸链占比分别为50.91％、3.64％、28.43％和17.02％.细胞转染试验结果显示,转染30 h后重组质粒组和空载体对照组均有荧光出现,且转染重组质粒的细胞内ABCD4基因表达量极显著高于空载体对照组(P＜0.01).实时荧光定量PCR结果显示,ABCD4基因在巴马小型猪皮下脂肪中表达量最高,显著高于其他组织(P＜0.05);在心脏中表达量最低.[结论]本试验成功扩增获得巴马小型猪ABCD4基因CDS区序列,ABCD4蛋白是一种疏水性跨膜蛋白,主要在巴马小型猪皮下脂肪中表达.本试验结果为探究A BCD4基因对猪背膘厚的影响提供了分子依据.

As one of the most critical economic traits, the litter performance of sows is influenced by their parity. Some studies have indicated a connection between the gut microbiota and the litter performance of animals. In this study, we examined litter performance in 1363 records of different parities of Large White sows. We observed a marked decline in TNB (Total Number Born) and NBH (Number of Healthy Born) We observed a marked decline in TNB (Total Number Born) and NBH (Number of Healthy Born) among sows with parity 7 or higher. To gain a deeper understanding of the potential role of gut microbiota in this phenomenon, we conducted 16S rRNA amplicon sequencing of fecal DNA from 263 Large White sows at different parities and compared the changes in their gut microbiota with increasing parity. The results revealed that in comparison to sows with a parity from one to six, sows with a parity of seven or higher exhibited decreased alpha diversity in their gut microbiota. There was an increased proportion of pathogenic bacteria (such as Enterobacteriaceae, Streptococcus, and Escherichia–Shigella) and a reduced proportion of SCFA-producing families (such as Ruminococcaceae), indicating signs of inflammatory aging. The decline in sow function may be one of the primary reasons for the reduction in their litter performance.

Abstract Background Bama miniature pigs aged between six (6 M) and twelve months (12 M) are usually used in human medical research as laboratory pigs. However, the difference in serum metabolic profiles from 6 to 12 M-old pigs remains unclear. This study aimed to identify the metabolic and physiological profiles present in the blood to further explain changes in Bama miniature pig growth. We collected blood samples from 6 M-, eight-month- (8 M-), ten-month- (10 M-), and 12 M-old healthy Guangxi Bama miniature pigs. A total of 20 blood physiological indices (BPIs) were measured: seven for white blood cells, eight for red blood cells, and five for platelet indices. Liquid chromatography and mass spectrometry-based non-targeted metabolomic approaches were used to analyze the difference in metabolites. The associations between the differences were calculated using Spearman correlations with Benjamini–Hochberg adjustment. The 100 most abundant differential metabolites were selected for analysis of their metabolic profiles. Results There were no significant differences in BPIs at different ages, but the mid cell ratio and red blood cell number increased with age. Seven BPIs in Bama miniature pigs were closer to human BPIs than to mouse BPIs. A total of 14 and 25 significant differential metabolites were identified in 6 M vs. 12 M and 8 M vs. 12 M, respectively. In total, 9 and 18 amino acids and their derivatives showed significantly lower concentrations in 6 M- and 8 M-old pigs than in 12 M-old pigs. They were identified as the core significantly different metabolites between the age groups 6 M vs. 12 M and 8 M vs. 12 M. Half of the enriched pathways were the amino acids metabolism pathways. The concentration of six amino acids (dl-tryptophan, phenylacetylglycine, muramic acid, N-acetylornithine, l(−)-pipecolinic acid, and creatine) and their derivatives increased with age. A total of 61 of the top 100 most abundant metabolites were annotated. The metabolic profiles contained 14 amino acids and derivatives, six bile acids and derivatives, 19 fatty acids and derivatives, and 22 others. The concentrations of fatty acids and derivatives were found to be inversely proportional to those of amino acids and derivatives. Conclusion These findings suggest high levels of MID cell ratio, red blood count, and amino acids in 12 M-old pigs as indicators for improved body function over time in Bama miniature pigs, similar to those in human development. This makes the pig a more suitable medical model organism than the mouse. The results of this study are limited to the characteristics of blood metabolism in the inbred Bama miniature pigs, and the effects of impacting factors such as breed, age, sex, health status and nutritional level should be considered when studying other pig populations.

Litter size is an important economic trait in pig production. However, the genetic mechanisms underlying varying litter size in Guangxi Bama Xiang pigs remain unknown. To identify selection signatures for litter size in Guangxi Bama Xiang pigs, we obtained 297 Illumina PorcineSNP50 BeadChip array data and the average born number (ABN) from parity one to nine in Guangxi Bama Xiang pigs. Fixation index (Fst) methods were used to identify the selection signature of the litter size, and three phenotypic gradient differential population pairs (according to the ABN) in individuals were used to reduce the false positives of signature selections. Single nucleotide polymorphisms (SNPs) were identified in the VEGFA promoter and exons. The general linear model was used to analyse the differences in distinct genotypes after they were typed using three-round multiplex PCR technology. Finally, the transcriptome factor and CpG island in the VEGFA promoter were predicted. A total of 328, 328 and 317 significant loci were identified in the 1st, 2nd and 3rd population pairs, respectively. After removing the false positives, 25 SNPs were defined as the selection signatures in relation to litter size. Ten (VEGFA, USP49, USP25, SRPK1, SLC26A8, RPL10A, PPARD, MAPK14, HMGA1 and CHRDL2) out of 52 genes in the selection regions were annotated as the candidate genes of litter size, respectively, VEGFA. There were no SNPs in the VEGFA exon region, but we obtained three SNPs (rs786889605, rs343769603 and rs323942424) in the VEGFA promoter regions. The ABN in CC was significantly higher than that in TT in rs786889605, and the ABN in TT was significantly lower than that in GG in rs323942424. Meanwhile, the mutation of the VEGFA promoter result in the loss of Sp1 and NF-1 and the formation of Oct-1. In summary, we obtained ten candidate genes, and two mutations in the VEGFA promoter that could be important potential molecular biomarkers for litter size in Bama Xiang pigs.

Lactobacillus delbrueckii subsp. bulgaricus (LDB) is an approved feed additive on the Chinese ‘Approved Feed Additives’ list. However, the possibility of LDB as an antibiotic replacement remains unclear. Particularly, the effect of LDB on microbiota and metabolites in the gastrointestinal tract (GIT) requires further explanation. This study aimed to identify the microbiota and metabolites present in fecal samples and investigate the relationship between the microbiota and metabolites to evaluate the potential of LDB as an antibiotic replacement in pig production. A total of 42 female growing-finishing pigs were randomly allocated into the antibiotic group (basal diet + 75 mg/kg aureomycin) and LDB (basal diet + 3.0 × 109 cfu/kg LDB) groups. Fecal samples were collected on days 0 and 30. Growth performance was recorded and assessed. 16S rRNA sequencing and liquid chromatography-mass spectrometry-based non-targeted metabolomics approaches were used to analyze the differences in microbiota and metabolites. Associations between the differences were calculated using Spearman correlations with the Benjamini–Hochberg adjustment. The LDB diet had no adverse effect on feed efficiency but slightly enhanced the average daily weight gain and average daily feed intake (p > 0.05). The diet supplemented with LDB increased Lactobacillus abundance and decreased that of Prevotellaceae_NK3B31_group spp. Dietary-supplemented LDB enhanced the concentrations of pyridoxine, tyramine, D-(+)-pyroglutamic acid, hypoxanthine, putrescine and 5-hydroxyindole-3-acetic acid and decreased the lithocholic acid concentration. The Lactobacillus networks (Lactobacillus, Peptococcus, Ruminococcaceae_UCG-004, Escherichia-Shigella, acetophenone, tyramine, putrescine, N-methylisopelletierine, N1-acetylspermine) and Prevotellaceae_NK3B31_group networks (Prevotellaceae_NK3B31_group, Treponema_2, monolaurin, penciclovir, N-(5-acetamidopentyl)acetamide, glycerol 3-phosphate) were the most important in the LDB effect on pig GIT health in our study. These findings indicate that LDB may regulate GIT function through the Lactobacillus and Prevotellaceae_NK3B31_group networks. However, our results were restrained to fecal samples of female growing-finishing pigs; gender, growth stages, breeds and other factors should be considered to comprehensively assess LDB as an antibiotic replacement in pig production.