Due to its high efficiency and safety, oocyte vitrification finds broad application in many fields of life sciences, such as clinical assisted reproduction and conservation of animal genetic resources. However, vitrification may cause cellular damage and reduce the quality of oocytes and their cumulus cells (CCs), which could be closely related to disorders in lipid metabolism. At present, the impact of vitrification upon the lipid profile of oocytes and CCs has not been systematically elucidated. In this study, we used porcine germinal vesicle cumulus–oocyte complexes (COCs) as a model to analyze their lipid characteristics after vitrification and in vitro maturation (IVM), utilizing untargeted lipid metabolomics. Our results showed that an overall count of 37 down-regulated and 8 up-regulated differential lipids was identified in the vitrified oocytes. Pathway analysis confirmed the enrichment in glycerophospholipid metabolism and fat digestion and absorption, etc. Combined with transcriptomic analysis, three enriched pathways were revealed, including the AMPK signaling pathway, metabolic pathways, and fatty acid elongation. On the other hand, a total of four down-regulated and eight up-regulated differential lipids were detected in the vitrified CCs. Pathway enrichment implicated autophagy, glycerophospholipid metabolism, etc. A joint analysis of metabolomic and transcriptomic data revealed four enrichment pathways, including cholesterol metabolism, fat digestion and absorption, regulation of lipolysis in adipocytes, and metabolic pathways. Notably, the supplementation of lysophosphatidylcholine during IVM attenuated oxidative stress, enhanced mitochondrial activity, and enhanced the viability and embryonic development of cryopreserved porcine oocytes. The results indicate that vitrification alters lipids in oocytes and CCs, and the supplementation of lipids plays a role in improving the quality of vitrified oocytes.

Pseudorabies virus (PRV) is a highly pathogenic agent that adversely impacts swine populations, leading to considerable economic losses within the Chinese pig industry. Furthermore, the potential for PRV to transmit across species from pigs to other hosts has attracted significant attention. In light of this, it is essential to identify efficacious antiviral agents for the treatment of PRV infection. In this study, we investigated the antiviral properties of Diltiazem HCl (DTZ) against PRV infection in susceptible cell lines. Our results demonstrated that DTZ significantly inhibited PRV infection in both PK15 and Vero cells. Moreover, this chemical compound exhibited antiviral activity against both variant and classical strains of PRV, as well as herpes simplex virus type 1 (HSV-1). Time-of-addition assays showed that DTZ exerted its inhibitory effect through specific interference with the virus replication process. Subsequent transcriptomic analysis via RNA sequencing indicated that the calcium signaling pathway might be involved in the antiviral properties of DTZ against PRV infection. Specifically, treatment with EGTA or calcium ion (Ca2+)-free medium inhibited PRV infection; this inhibitory effect was substantially mitigated upon the reintroduction of CaCl2. In summary, DTZ effectively suppressed PRV infection in vitro, demonstrating its potential as an antiviral agent against PRV infections.

The preservation of porcine oocytes is critically important for advancing superior breeds and conserving genetic resources in pig production. Vitrification has gained traction as a preferred alternative to slow freezing for porcine oocytes because of its effectiveness in reducing ice crystal formation, yet it can still negatively affect oocyte quality, compromising their in vitro maturation (IVM) and later embryonic development. Long non-coding RNAs (lncRNAs) have proven to be key players in numerous biological processes, such as oocyte growth, maturation, and early embryogenesis. Despite this, the effects of vitrified porcine germinal vesicle (GV) oocytes, particularly regarding IVM and the dynamic expression patterns of lncRNAs during embryonic development, remain largely unclear. To address this gap, this study conducted lncRNA sequencing at the metaphase II (MII), parthenogenetic 4-cell embryo, and parthenogenetic blastocyst stages sourced from both fresh and vitrified GV oocytes. This method enabled us to ascertain the impact of vitrification on lncRNA expression throughout oocyte maturation and embryonic development. Results identified 773 differentially expressed lncRNAs (DELs) at the MII stage, 1973 at the parthenogenetic 4-cell, and 1192 at the parthenogenetic blastocyst. Enrichment analysis of forecasted target genes revealed their involvement in key regulatory pathways associated with the cell cycle, meiosis, stress response, and metabolic activity. Overall, this study provides a comprehensive overview of lncRNA expression during oocyte maturation and embryonic development following porcine GV oocyte vitrification, thereby shedding light on the molecular mechanisms behind vitrification-induced damage.

Vitrification of porcine immature oocytes at the germinal vesicle (GV) stage reduces subsequent embryo yield and changes at the molecular level may occur during embryonic development. Therefore, the present study used porcine parthenogenetic embryos as a model to investigate the effect of GV oocyte vitrification on the transcriptional profiles of the resultant embryos at the 4-cell and blastocyst stages using the Smart-seq2 RNA-seq technique. We identified 743 (420 up-regulated and 323 down-regulated) and 994 (554 up-regulated and 440 down-regulated) differentially expressed genes (DEGs) from 4-cell embryos and blastocysts derived from vitrified GV oocytes, respectively. Functional enrichment analysis of DEGs in 4-cell embryos showed that vitrification of GV oocytes influenced regulatory mechanisms related to transcription regulation, apoptotic process, metabolism and key pathways such as the MAPK signaling pathway. Moreover, DEGs in blastocysts produced from vitrified GV oocytes were enriched in critical biological functions including cell adhesion, cell migration, AMPK signaling pathway, GnRH signaling pathway and so on. In addition, the transcriptomic analysis and quantitative real-time PCR results were consistent. In summary, the present study revealed that the vitrification of porcine GV oocytes could alter gene expression patterns during subsequent embryonic developmental stages, potentially affecting their developmental competence.

为从全基因组水平探索影响藏猪骨骼肌发育的遗传变异,本研究对5头迪庆藏猪进行全基因组重测序,并合并NCBI数据库中来自3个省的藏猪、与藏猪同样小体型及生长慢的巴马香猪、及体型大且生长快速的杜洛克和大白猪共70个猪只的全基因组重测序数据进行整合分析,获得全基因组SNP后结合选择信号检测与等位基因频率(AF)分析鉴定藏猪-香猪与杜洛克-大白猪的遗传差异;利用GO和KEGG功能富集分析藏猪骨骼肌转录组与蛋白组相关基因.结果发现:1)2 211个基因在藏猪-香猪与杜洛克-大白猪2个比较组中存在遗传差异,138个基因富集到骨骼肌发育相关的GO功能集及KEGG通路;2)9个基因(UCHL3、POSTN、COL12A1、PGK1、JPH1、GPT2、RBFOX1、FLNB和DCX)已报道参与藏猪60 d胚胎背最长肌发育调控,1个(CKM)参与6月龄藏猪背最长肌发育调控;3)10个基因共包含936个SNP,其中655个SNP位于基因间区,255个是内含子变异,少量SNP是外显子及调控区变异.本研究从全基因组水平鉴定了与藏猪骨骼肌发育相关基因及其遗传变异,为以后持续深入解析藏猪骨骼肌发育的遗传机制提供参考.

Antibody level detection is an important indicator for effectively evaluating animal epidemic prevention work,and strengthening the monitoring of antibody levels of certain animal diseases in the jurisdiction can provide a scientific basis for for-mulating animal disease immunization procedures and prevention and control measures.In order to understand the immune anti-body situation of foot-and-mouth disease in Zhaotong City from 2019 to 2022,the ELISA method was used to detect the lev-el of immune antibodies to foot-and-mouth disease O and A viruses in a total of 17210 sheep serum in nine counties and dis-tricts of Zhaotong City over a four-year period,and compared the immunization effects of foot-and-mouth disease in sheep in different counties and different years.In the nine counties and districts where samples were collected,the foot-and-mouth disease O-type antibodies were higher than the national level(＞70％);The antibody level of foot-and-mouth disease type A antibody in Sui-jiang County and Yiliang County did not reach the national level(＞70％).In Zhaotong City,the qualification rate of immune antibodies to FMD O in Zhaotong City has generally met the require-ments stipulated by the state,but the antibody level in some counties and districts is still at a rela-tively low level,and the immunization program should be adjusted on the original immunization program to achieve a better immunization level.The antibody levels of foot-and-mouth disease type A did not reach the national level in two coun-ties and districts,and the rest of the counties and districts met the national standard.Their immuni-zation schedules should be improved or more pro-tective vaccines replaced to increase levels of FMD type A antibodies in flocks and avoid economic los-ses due to foot-and-mouth disease type A virus infection.

[目的]研究胎牛血清(fetal bovine serum,FBS)在猪孤雌囊胚玻璃化冷冻后恢复培养中的作用.[方法]本试验以体外培养第5天的猪孤雌激活囊胚为材料,将新鲜和冷冻囊胚分别在含10％FBS(V/V)的胚胎培养液中继续培养48 h,即分为新鲜组(Fresh)、新鲜+FBS组(Fresh+FBS)、冷冻组(Vitrified)、冷冻+FBS组(Vitrified+FBS).观察各组囊胚的扩张和孵化能力,检测胚胎的细胞膜损伤、凋亡细胞数目、总细胞数目、胞内活性氧(ROS)水平、线粒体活性以及发育相关基因的表达水平.[结果]与Fresh和Vitrified组相比,Fresh+FBS和Vitrified+FBS组的完全扩张率、孵化率和囊胚细胞总数均显著提高(P＜0.05),细胞膜损伤率和细胞凋亡率均显著降低(P＜0.05).与 Fresh 组相比,Vitrified 组 ROS 水平显著升高(P＜0.05),Fresh+FBS 和 Vitrified+FBS 组 ROS 水平均显著降低(P＜0.05).Vitrified+FBS组的线粒体活性显著高于Vitrified组(P＜0.05)、显著低于Fresh+FBS组(P＜0.05),与Fresh组无显著差异(P＞0.05).相比于Fresh组,Vitrified组POU结构域与类转录因子1(POU5F1)表达水平显著上升(P＜0.05)、过氧化氢酶(CAT)表达水平显著下降(P＜0.05).增殖细胞核抗原(PCNA)、DNA甲基转移酶3A(DNMT3A)、超氧化物歧化酶1(SOD1)、BCL2相关X蛋白(BAX)/BCL2L1的表达水平在Fresh和Vitrified组之间均无显著性差异(P＞0.05).Fresh+FBS和Vitrified+FBS组中PCNA、SOD1和CAT的表达水平显著高于Fresh和Vitrified组(P＜0.05).[结论]体外培养第5天的新鲜和冷冻猪孤雌囊胚在10％FBS中继续培养,其发育能力及胚胎质量均得到明显改善.

[目的]从全基因组水平解析藏猪与大白猪间的遗传差异,并鉴定出与藏猪脂质沉积相关的基因,为藏猪优质肉质特征的利用及新品种培育提供理论参考.[方法]选用5份自测云南迪庆藏猪的全基因组重测序数据,以及NCBI数据库中来自西藏、四川、甘肃地区34份藏猪和具有低脂肪沉积特征的14份大白猪全基因组重测序数据,获得全基因组SNP后以VCFtools进行选择信号检测,采用VCFtools分别计算候选SNP在藏猪和大白猪中的等位基因频率,再利用自编Python脚本及所有SNP注释文件进行注释以确定候选基因,最后运用DAVID对候选基因进行GO功能注释和KEGG信号通路富集分析,进一步鉴定与脂质沉积相关的功能基因.[结果]经全基因组重测序及与参考基因组比对、过滤后得到19374832个高质量的SNPs,根据受选择区域条件筛选出370778个候选SNPs,其中符合在藏猪中等位基因频率＞0.5而在大白猪中等位基因频率＜0.5条件的SNPs有152877个,共注释到1486个候选基因.GO功能注释分析结果显示,1486个候选基因注释到17个GO功能簇(Clusterl～Cluster17),其中Cluster11和Cluster12与脂肪生成相关,涉及与脂滴转运、结合过程相关的基因NME4、C4BPA、AP2M1、CD36、FABP5、PMP2、PDZD8、OSBPL6、APOD和OSBPL1A,与脂肪酸合成、延伸和代谢过程相关的基因TECR、ELOVL7、HACD2,以及与脂肪形成相关的细胞外基质基因(CCN3、EGFL6、KAZALD1、ASPN、ADAMTS18、ECM2、COL18A1、COL3A1和COL4A5);通过文献分析还发现 8 个基因(APOOL、PLAG1、PLAUR、SUCLG2、CPE、AKR1B1、SLC4A4和CHPT1)在藏猪与大白猪皮下脂肪或肌肉组织呈差异表达,且与脂肪形成相关.[结论]藏猪与大白猪间的脂肪沉积差异涉及脂肪形成的多个过程和多个基因,细胞外基质基因也可能与藏猪脂肪形成相关,且这些基因在藏猪中均具有较高的等位基因频率.

研究采用测序技术对云南地方猪杂交后代脂肪酸转运蛋白1(FATP1)基因进行了多态性遗传分析.结果显示:在云南地方猪杂交后代FATP1基因第一外显子上存在1个SNP(c.70 C>T)位点,该位点在杜藏猪(杜洛克×迪庆藏猪)群体中无多态性(仅CC型),在杜小猪(杜洛克×滇南小耳猪)群体和杜小藏猪(杜洛克×滇南小耳猪×迪庆藏猪)群体中存在多态性(CC型和CT型),均处于Hardy-Weinderg平衡(P>0.05),且属低度多态(He<0.25,PIC<0.25).

为了解昆明滇池红嘴鸥(Larus ridibundus)携带的沙门氏菌(Salmonella spp.)的耐药性、毒力特征及致病性等情况,2019年11月,从昆明滇池随机选择100只越冬红嘴鸥,对其泄殖腔内容物进行细菌分离培养、生化鉴定、染色镜检、分子生物学鉴定、多位点序列分型和血清学鉴定;采用药敏纸片琼脂法对5大类14种抗菌药物进行耐药检测,使用PCR扩增检测耐药基因,分析耐药情况;通过毒力基因检测及动物致病性试验对所分离的细菌进行致病性分析.结果显示:确定其为ST19型鼠伤寒沙门氏菌(Salmonella Typhimurium),抗原结构式为O抗原1,4,[5],12;H抗原i,1,2,主要对阿莫西林、环丙沙星、四环素和多西环素耐药,且检出四环素类耐药基因tetB,喹诺酮类耐药基因gyrA、gyrB、parC和psrE.该分离菌检测出mogA、sseL、mgtC、bcfA、araB、spvC、spvB、stn、fimA、orgA、sopB、sipB和spaN 13种毒力基因.动物试验结果表明,该分离菌株对试验小鼠有一定致病性,接种浓度为5×106、5×107、5×108 cfu/mL的小鼠死亡率分别为60％、100％和100％,死亡小鼠出现肝脏肿大、肺部局部充血和肠壁变薄等病变.

细胞是机体最基本的结构组成及功能单位,细胞类型和功能由其整个转录表达谱决定,通过单细胞转录组测序可以获得单个细胞转录表达谱,由此以高精度分辨率鉴定细胞类型、细胞状态以及稀有类型细胞,从而可以在单细胞水平分析细胞动态变化及细胞间的关系,深入解析驱动细胞变化及细胞异常背后的分子细胞机制.随着单细胞测序技术稳定性和测序通量的提高,以及测序成本的降低,其在发育生物学、肿瘤、免疫及疾病等领域被广泛应用,研究对象主要涉及人及模式生物,在动物上的应用研究相对较少.本文主要介绍单细胞转录组测序技术及其生物学应用并综述目前其在动物上的一些开创性研究,以期为今后更好的在动物上应用单细胞转录组测序提供方法参考.

研究通过测序技术对迪庆藏猪脂肪分化蛋白基因(ADFP)部分启动子区域和外显子1进行了多态位点遗传分析.结果发现,迪庆藏猪在ADFP基因启动子区域存在2个SNPs,外显子1上存在6个SNPs;各SNPs均处于Hardy-Weinberg平衡,多态信息含量和杂合度均小于0.5.研究了解了迪庆藏猪ADFP基因的变异情况及群体遗传特征,为下一步开展脂肪沉积基因的遗传育种奠定了理论基础.

为了研究迪庆藏猪杂交后代的生长发育规律,同时更好地指导生产和管理,对0～6月龄的迪庆藏猪杂交后代A品系进行体重测定,采用Logistic和Gompertz两种非线性曲线模型,数据通过SPSS 17.0软件进行拟合分析.结果表明:Logistic和Gompertz模型的拟合度均在0.99以上,都能够较好地拟合迪庆藏猪杂交后代A品系体重的生长曲线,拟合值与实际生长值基本吻合,通过对比发现Logistic模型拟合效果更好.Logistic模型能较好地拟合迪庆藏猪杂交后代的生长曲线,拟合的拐点体重和拐点日龄分别为71.93 kg和180.11 d.

The dynamic changes in protein expression are well known to be required for oocyte meiotic maturation. Although proteomic analysis has been performed in porcine oocytes during in vitro maturation, there is still no full data because of the technical limitations at that time. Here, a novel tandem mass tag (TMT)-based quantitative approach was used to compare the proteomic profiles of porcine immature and in vitro mature oocytes. The results of our study showed that there were 763 proteins considered with significant difference-450 over-expressed and 313 under-expressed proteins. The GO and KEGG analyses revealed multiple regulatory mechanisms of oocyte nuclear and cytoplasmic maturation such as spindle and chromosome configurations, cytoskeletal reconstruction, epigenetic modifications, energy metabolism, signal transduction and others. In addition, 12 proteins identified with high-confidence peptide and related to oocyte maturation were quantified by a parallel reaction monitoring technique to validate the reliability of TMT results. In conclusion, we provided a detailed proteomics dataset to enrich the understanding of molecular characteristics underlying porcine oocyte maturation in vitro.

Cryopreservation of embryos has been confirmed to cause oxidative stress as a factor responsible for impaired developmental competence. Currently, astaxanthin (Ax) raises considerable interest as a strong exogenous antioxidant and for its potential in reproductive biology. The present study aimed to investigate the beneficial effects of Ax supplementation during in vitro culture of vitrified porcine zygotes and the possible underlying mechanisms. First, the parthenogenetic zygotes were submitted to vitrification and then cultured in the medium added with various concentrations of Ax (0, 0.5, 1.5, and 2.5 μM). Supplementation of 1.5 μM Ax achieved the highest blastocyst yield and was considered as the optimal concentration. This concentration also improved the blastocyst formation rate of vitrified cloned zygotes. Moreover, the vitrified parthenogenetic zygotes cultured with Ax exhibited significantly increased mRNA expression of CDX2, SOD2, and GPX4 in their blastocysts. We further analyzed oxidative stress, mitochondrial and lysosomal function in the 4-cell embryos and blastocysts derived from parthenogenetic zygotes. For the 4-cell embryos, vitrification disturbed the levels of reactive oxygen species (ROS) and glutathione (GSH), and the activities of mitochondria, lysosome and cathepsin B, and Ax supplementation could fully or partially rescue these values. The blastocysts obtained from vitrified zygotes showed significantly reduced ATP content and elevated cathepsin B activity, which also was recovered by Ax supplementation. There were no significant differences in other parameters mentioned above for the resultant blastocysts. Furthermore, the addition of Ax significantly enhanced mitochondrial activity and reduced lysosomal activity in resultant blastocysts. In conclusion, these findings revealed that Ax supplementation during the culture period improved subsequent embryonic development and quality of porcine zygotes after vitrification and might be used to ameliorate the recovery culture condition for vitrified embryos.

The change of sperm protein profile after the cryopreservation process may influence fertilization and early embryonic development. The purpose of the present study was to identify ram sperm proteomic modifications induced by the cryopreservation process using the isobaric tags for relative and absolute quantification labeling technology (iTRAQ) coupled with the parallel reaction monitoring (PRM) technology. Semen samples were collected from five Yunnan semi-fine wool rams using an electroejaculator. Sperm motility (CASA), plasma membrane (HOST test), and acrosome integrity (FITC-PSA) were evaluated after freeze-thawing. The total proteins of fresh and frozen-thawed sperm were extracted and purified, followed by identifying ram sperm proteomic modifications using the isobaric tags for relative and absolute quantification labeling technique (iTRAQ) coupled with the parallel reaction monitoring (PRM) technology. The results showed a significant reduction (P < 0.05) in all sperm parameters after thawing. 126 differentially abundant proteins (DAPs) were identified through comparison of the proteomes between fresh and frozen-thawed sperm. Among them, 90 proteins were down-regulated after the cryopreservation process. The remaining 36 proteins were up-regulated in frozen-thawed sperm. The results of functional annotation demonstrated the potential relationship of 10 DAPs with oxidoreductase activity. 18 and 15 DAPs may be involved in the stress and carbohydrate metabolic process, respectively. Furthermore, 8 DAPs may be functionally associated with reproduction. The Kyoto Encyclopedia of Genes and Genomes (KEGG) results demonstrated the primary enrichment of these identified DAPs in metabolic activities, disease, and oxidative phosphorylation. In order to confirm the reliability of the iTRAQ results, the changing trends of 10 proteins analyzed by PRM were similar to those of the corresponding proteins identified by iTRAQ. In conclusion, the cryopreservation process modifies the proteome of ram sperm, possibly leading to compromised fertility of post-thaw sperm. Additionally, the identified DAPs in this study may function as potential biomarkers for assessing the post-thaw quality of ram semen.

In this study, the complete proteome of goat ejaculated semen including spermatozoa and seminal plasma was established, applying a tandem mass tag (TMT) labeling together with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In seminal plasma, 2299 proteins were identified and 2098 proteins were quantified. The GO analysis demonstrated that 32% proteins were involved in metabolic activities. 46% proteins are located at intracellular region, intracellular organelle, and membrane-bounded organelle. Regarding molecular function, 40% proteins are engaged on protein binding, hydrolase activity, and ion binding. The KEGG analysis indicated a primary involvement of the identified proteins in protein processing in endoplasmic reticulum, lysosome, and proteome. In spermatozoa, 2491 proteins were identified and quantified. 39% proteins are involved in metabolic activities. 48% proteins are located at intracellular region, intracellular organelle, and membrane-bounded organelle. 38% proteins are engaged on protein binding, hydrolase activity, and ion binding. The KEGG analysis demonstrated their roles derived from the identified proteins in proteasome, glycolysis, pyruvate metabolism, and citrate cycle. Additionally, 1312 proteins were simultaneously presented in spermatozoa and seminal plasma. The involvement of 42% proteins in metabolic activities were observed. 47% proteins are located at intracellular region, intracellular organelle, and membrane-bounded organelle. The common proteins are mainly engaged on protein processing in endoplasmic reticulum, proteome, glycolysis, lysosome, and citrate cycle. Collectively, this study established the protein database of goat semen. More studies should be used to elucidate functionality of these identified proteins.

[目的]从分子遗传学角度分析不同基因型迪庆藏猪群体不同组织中ADFP基因的表达差异,为迪庆藏猪的选育工作提供参考依据.[方法]利用PCR扩增测序方法对113头迪庆藏猪ADFP基因进行扩增测序,利用DNA-STAR 5.0对序列进行比对分析及多态性位点(SNPs)检测;同时利用实时荧光定量PCR方法对迪庆藏猪不同组织中ADFP基因相对表达量进行检测,并运用SPSS 20.0分析ADFP基因多态性对迪庆藏猪群体不同组织中ADFP基因表达水平的影响.[结果]在迪庆藏猪ADFP基因外显子6上发现3个SNPs,分别为SNP1(C→T)、SNP2(C→T)和SNP3(T→C),其中SNP1和SNP2位点属于低度多态(PIC/He<0.25),SNP3位点属于中度多态(0.25<PIC/He<0.5).迪庆藏猪不同组织中ADFP基因相对表达量存在显著差异(P<0.05,下同),皮下脂肪中相对表达量最高,其次为肺脏、肝脏、脾脏、肾脏、腿肌和背肌,心脏中相对表达量最少;另外,ADFP基因SNPs不同基因型迪庆藏猪群体中各组织的相对表达量也存在显著差异,尤其在SNP2位点杂合基因型群体肾脏中相对表达量显著高于纯合基因型群体肾脏中相对表达量,表明ADFP基因SNPs变异对迪庆藏猪组织中ADFP基因表达具有影响.[结论]ADFP基因SNPs变异对迪庆藏猪组织中ADFP基因会产生差异表达,进而影响机体脂肪沉积,ADFP基因可作为迪庆藏猪肌内脂肪沉积的候选基因.

[目的]探究复合益生菌对超早期断奶(7d)杜藏乳仔猪肠道微生物多样性及物种丰度的影响,以减轻乳仔猪断奶应激,为微生物饲料添加剂的研发提供科学依据.[方法]选取7日龄杜藏乳仔猪30头,随机分为3组,每组10头.对照组(ZM组)随母猪哺乳,试验I组(ZD组)哺喂代乳粉,试验II组(ZY组)哺喂代乳粉+复合益生菌,试验周期21 d.试验结束当天(28日龄)收集乳仔猪粪便,采用Illumina高通量测序分析粪便样品的菌群结构组成.[结果]Illumina高通量测序获得ZM组、ZY组和ZD组杜藏乳仔猪粪便样品共有OTU为327个,ZM组的特有OTU为247个,ZD组的特有OTU为84个,ZY组的特有OTU为96个.在门分类水平上,ZM组杜藏乳仔猪粪便样品中相对丰度最高的菌门为厚壁菌门,ZY组和ZD组为拟杆菌门;ZY组和ZD组的厚壁菌门/拟杆菌门比值分别为0.77和0.92,较ZM组(1.76)分别下降56.25％和47.73％.在属分类水平上,ZD组和ZY组杜藏乳仔猪粪便样品中的优势菌属为普氏菌属_9,ZM组则为乳杆菌属;ZD组杜藏乳仔猪粪便中布劳特氏菌属、肠球菌属和吉氏副拟杆菌属的相对丰度较其他2个处理组显著上升(P<0.05,下同);而ZY组杜藏乳仔猪粪便中普雷沃氏菌科_NK3B31群、普雷沃氏菌科_UCG-003、厌氧弧菌属、罕见小球菌属、瘤胃菌科_NK4A214群、瘤胃菌科_UCG-005、未明确普雷沃氏菌科及Family_XIII_AD3011_group的相对丰度显著高于ZD组.[结论]在杜藏超早期断奶仔猪代乳粉中添加复合益生菌能有效提高其肠道菌群结构多样性,同时提高与碳水化合物代谢和产短链脂肪酸相关菌群的相对丰度,即复合益生菌具有潜在促进营养物质代谢和抗炎症的功能,可缩短消化道微生物区系由哺乳型向饲料型的转变历程.

[背景]随着"禁抗令"的推行,如何寻找安全的抗生素替代品迫在眉睫.[目的]研究戊糖片球菌368对育肥猪的影响.[方法]选用20头育肥猪,随机分为2组,每组2个重复,每个重复5头.对照组饲喂基础饲粮,试验组饲喂基础饲粮+戊糖片球菌368(1010CFU/kg),试验期28 d.[结果]试验期内,试验组和对照组的平均日增重、料重比和平均日采食量均无显著差异(P＞0.05).根据OTU聚类分析,12个样品共获得1 036个OTU,聚类为17个门23个纲37个目67个科199个属350个种.主坐标分析(Principal Co-Ordinates Analysis,PCoA)发现2组样品显著分开.在门水平上,螺旋体门的相对丰富极显著上升(P＜0.01);在属水平上,狭义梭菌属和土孢菌属的相对丰度显著上升(P＜0.05),链球菌属(P＜0.01)、毛螺菌科_XPB1014群(P＜0.05)、Norank_f_p-251-o5(P＜0.01)和瘤胃球菌科_NK4A214群(P＜0.05)的相对丰度下降.多级物种差异判别分析(LDA Effect Size,LEfSe)发现,LDA值大于2.0时,获得60个生物标记物,其中,P组为31个;C组内LDA值最大的是链球菌属,P 组最大的是梭菌目.偏最小二乘判别分析法(Partial Least Squares Discrimination Analysis,PLS-DA)发现,正负离子模式下,样品完全分开且差异显著(P＜0.05),P组中白喉酰胺、2-羟基十四烷酸、Furanofukinin、多巴醌、Avocadene、N-(1-脱氧-1-果糖基)脯氨酸和N-(1-脱氧-1-果糖基)甘氨酸是显著(P＜0.05)或极显著(P＜0.01)上调物质,ArtoninU、1α,25-二羟基-9,11-麦角酸-3-脱氧-维生素D3和24-脱氧维生素D3则是显著(P＜0.05)或极显著(P＜0.01)下调物质.[结论]添加戊糖片球菌368后能改善猪粪便菌群的结构,提高抗病菌(狭义梭菌属和土孢菌属)并降低条件致病菌(链球菌属)的相对丰度,提高风味物质前体[N-(1-脱氧-1-果糖基)脯氨酸和N-(1-脱氧-1-果糖基)甘氨酸]的含量,降低维生素D3(24-脱氧维生素D3和1α,25-二羟基-9,11-麦角酸-3-脱氧-维生素D3)的含量,具有促进健康和抗病的潜在功能.