Effect of hay type on cecal and fecal microbiome and fermentation parameters in horses

The effect of hay type on the microbiome of the equine gastrointestinal tract is relatively unexplored. Our objective was to characterize the cecal and fecal microbiome of mature horses consuming alfalfa or Smooth Bromegrass (brome) hay. Six cecally cannulated horses were used in a split-plot design run as a crossover in two periods. The whole plot treatment was ad libitum access to brome or alfalfa hay fed over two 21-d acclimation periods with subplots of sampling location (cecum and rectum) and sampling hour. Each acclimation period was followed by a 24-h collection period where cecal and fecal samples were collected every 3 h for analysis of pH and volatile fatty acids (VFA). Fecal and cecal samples were pooled and sent to a commercial lab (MR DNA, Shallowater, TX) for the amplification of the V4 region of the 16S rRNA gene and sequenced using Illumina HiSeq. The main effects of hay on VFA, pH, and taxonomic abundances were analyzed using the MIXED procedure of SAS 9.4 with fixed effects of hay, hour, location, period, and all possible interactions and random effect of horse. Alpha and beta diversities were analyzed using the R Dame package. Horses fed alfalfa had greater fecal than cecal pH (P ≤ 0.05), whereas horses fed brome had greater cecal than fecal pH (P ≤ 0.05). Regardless of hay type, total VFA concentrations were greater (P ≤ 0.05) in the cecum than in feces, and alfalfa resulted in greater (P ≤ 0.05) VFA concentrations than brome in both sampling locations. Alpha diversity was greater (P ≤ 0.05) in fecal compared with cecal samples. Microbial community structure within each sampling location and hay type differed from one another (P ≤ 0.05). Bacteroidetes were greater (P ≤ 0.05) in the cecum compared with the rectum, regardless of hay type. Firmicutes and Firmicutes:Bacteroidetes were greater (P ≤ 0.05) in the feces compared with cecal samples of alfalfa-fed horses. In all, fermentation parameters and bacterial abundances were impacted by hay type and sampling location in the hindgut.     

参考群筛选方法及规模对基因型填充准确性的影响

为探究基于A矩阵期望遗传关系最大化（maximizing the expected genetic relationship for matrix A,RELA）、基于A矩阵目标群体遗传方差最小化（minimized the target population genetic variance for matrix A,MCA）、平均亲缘关系最大化（the highest mean kinship coefficients,KIN）、随机选择（random selection,RAN）、共同祖先筛选（common ancestor,CA）等不同参考群筛选方法及参考群规模对基因型填充准确性的影响。本研究使用矮小型黄羽肉鸡作为试验群体,采用鸡600K SNP芯片（Affymetrix Axion HD genotyping array）进行基因分型,测定435羽子代公鸡45、56、70、84、91日龄体重。利用Beagle软件将低密度SNP芯片填充为高密度SNP芯片数据,比较不同参考群筛选方法、参考群规模对基因型填充准确性的影响,以及填充芯片基因组预测准确性。结果表明,使用Beagle 4.0结合系谱信息进行填充效果最佳,其次为Beagle 4.0,而Beagle 5.1填充效果最差。使用MCA方法筛选参考群进行基因型填充准确性最高,使用RAN方法筛选参考群进行基因型填充准确性最低,MCA、RELA、CA 3种方法基因型填充准确性差别较小。相比其他方法,使用MCA方法筛选个体作为参考群将低密度SNP芯片填充至高密度SNP芯片进行基因组选择的预测准确性较高,与真实高密度SNP芯片的基因组预测准确性相差甚微。随着参考群规模增大,基因型填充准确性也随之增加,但增速逐渐下降,最后趋于平缓。综上所述,可以通过参考群筛选方法构建参考群以及控制参考群规模,以保证基因型填充和基因组预测准确性并节省成本,本研究为基因型填充在畜禽遗传育种中的应用提供技术参考。     

内生真菌对高寒草地紫花针茅凋落物分解的影响

禾草内生真菌对于提高宿主植物对生物和非生物胁迫的耐性发挥着积极作用,然而,禾草内生真菌共生体凋落物分解对于草地生态系统养分循环的影响仍不清楚。本研究以紫花针茅为研究对象,比较带内生真菌（E+）与不带内生真菌（E-）植株凋落物分解过程中重量及全氮、木质素和纤维素含量,以及木质素∶N、纤维素∶N与全氮、木质素和纤维素残留率的变化,以期揭示内生真菌在紫花针茅凋落物分解过程中发挥的作用。主要结果如下：E+紫花针茅凋落物分解速率高于E-,具有更短的分解周期;随着时间的延长,紫花针茅凋落物木质素含量和木质素∶N由E+显著高于E-逐渐变为二者之间差异不显著,纤维素含量、纤维素∶N和纤维素残留率则逐渐变为E+显著低于E-;另外,随着分解时间延长,E+和E-凋落物全氮含量呈上升趋势,而全氮残留率呈现出下降-上升-下降的趋势,且后期E+显著低于E-。因此,内生真菌不同程度地促进了紫花针茅凋落物全氮、木质素和纤维素的降解。     

三种荒漠植物叶绿素荧光参数日变化特征

利用Photosynq MultiseQ多功能植物测量仪测定塔克拉玛干沙漠腹地3种主要防护林植物头状沙拐枣、多枝柽柳和梭梭的叶绿素荧光参数日变化,探讨其与环境因子的关系,以揭示3种植物对极端环境的适应策略,为沙漠公路防护林的管护提供理论依据。结果表明：1）3种土壤都呈碱性;0~30 cm土层中的电导率（EC）远高于30 cm以下的土层。1 m内梭梭土层中的含水量高于多枝柽柳高于头状沙拐枣。2）3种植物的光合有效辐射（PAR）、叶片温度和线性电子传递（LEF）及植物周围的大气温度日变化均先升后降,且都在14：00达到最高;而大气湿度、实际光化学效率Y_（II）日变化均呈‘V’型。3）PAR基本上与3种植物的LEF、非光化学淬灭系数（NPQ）、调节性能量耗散量子产量Y_（NPQ）、非调节性能量耗散量子产量Y_（NO）呈显著正相关,与实际光化学效率Y_（II）呈显著负相关;3种植物的Y_（II）与Y_（NPQ）、Y_（NO） 均呈极显著负相关。3种植物的LEF、Y_（II）、Y_（NPQ）的日均值差异不显著,但梭梭与多枝柽柳的NPQ和Y_（NO）日均值差异显著。4）在3种植物的叶绿素荧光参数中,梭梭和头状沙拐枣的LEF、NPQ和Y_（II）要高于多枝柽柳;而多枝柽柳的Y_（NO）要显著高于头状沙拐枣和梭梭。因此,高温和高光均对3种植物造成了不同程度的影响,通过主成分分析表明,3种植物的抗逆性强弱顺序分别为梭梭>头状沙拐枣>多枝柽柳。     

猪流行性腹泻病毒S1D蛋白的优化表达及其单克隆抗体的制备

为进一步探究猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)S蛋白的抗原表位及其功能,本试验通过优化S1D基因的密码子,构建了S1D基因未优化的重组原核表达质粒pET-S1D和已优化的pET-ΔS1D,并进行了诱导表达和纯化。使用SDS-PAGE和Western blotting方法验证S1D、ΔS1D蛋白在大肠杆菌内得到正确表达,利用Image J软件对S1D、ΔS1D蛋白表达量进行灰度扫描,通过t检验分析两者差异性。将纯化的ΔS1D 蛋白免疫 BALB/c小鼠,通过细胞融合、筛选及亚克隆,获得单克隆细胞株。利用体内诱生法制备抗PEDV S1D蛋白的单克隆抗体腹水,使用ELISA、Western blotting、间接免疫荧光试验3种方法对腹水效价及特异性进行检测和验证。SDS-PAGE和Western blotting结果显示,表达S1D、ΔS1D蛋白的样品均在34 ku处出现正确的目的条带。t检验结果表明, S1D、ΔS1D两者蛋白表达量差异极显著(P<0.01)。ELISA结果显示, 腹水的抗体效价达到了1∶1 000 000,腹水与PEDV病毒粒子和纯化后的ΔS1D蛋白反应均呈阳性,与PEDV N蛋白、pET-32a(+)空载体蛋白和猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)、猪传染性胃肠炎病毒(Transmissible gastroenteritis virus,TGEV)、猪瘟病毒(Classical swine fever virus,CSFV)、猪德尔塔冠状病毒(Porcine deltacoronavirus,PDCoV)和猪急性腹泻综合征冠状病毒(Swine acute diarrhea syndrome coronavirus,SADS-CoV) 5种病毒反应均呈阴性。Western blotting结果显示,腹水与ΔS1D蛋白和PEDV S蛋白分别在34和180 ku处有特异性条带出现,与pET-32a(+)空载体蛋白、正常Vero细胞蛋白均无特异性条带出现。间接免疫荧光试验结果显示,腹水及阳性对照组均能使细胞出现特异性绿色荧光信号,而空白及阴性对照组均未见绿色荧光信号。密码子优化可在原核表达系统中显著提高重组蛋白的表达水平,本研究基于高效表达的ΔS1D蛋白,成功制备了1株能稳定分泌与 PEDV S蛋白特异性结合的单克隆抗体的细胞株,为进一步探究 PEDV S蛋白抗原表位及蛋白功能的研究奠定了基础。     

松辽黑猪METTL23基因多态性及其与繁殖性状的关联分析

为探究松辽黑猪METTL23基因多态性及其与繁殖性状的关联性,试验选取178头松辽黑猪母猪为研究对象,利用Sanger直接测序法查找METTL23基因外显子1~5的单核苷酸多态性(SNP),使用SPSS 19.0软件分析METTL23基因SNP位点与松辽黑猪繁殖性状(总产仔数、产活仔数、断奶仔猪数、初生重、3周龄重、断奶重、乳头数)的关联性。结果显示,仅在松辽黑猪METTL23基因外显子4发现1个SNP位点,命名为A62G,在A62G位点上检测到3种基因型,分别是AA、AG和GG。GG和G分别为优势基因型和优势等位基因,且A62G位点处于Hardy-Weinberg平衡状态(P>0.05)。该位点的遗传纯合度(Ho)为0.6516,高于遗传杂合度(He,0.3484),说明其在松辽黑猪群体中的变异较小;多态信息含量(PIC)为0.2877,属于中度多态位点(0.25<PIC<0.5),说明该遗传标记能够提供一定量的遗传信息。关联分析结果表明,GG基因型个体总产仔数、产活仔数和断奶仔猪数均显著或极显著高于AA基因型个体(P<0.05;P<0.01),总产仔数还显著高于AG基因型个体(P<0.05);AG基因型个体断奶仔猪数显著高于AA基因型个体(P<0.05);初生重、3周龄重、断奶重、乳头数各基因型间差异均不显著(P>0.05)。综上所述,METTL23基因外显子4存在多态性位点A62G,该位点与松辽黑猪产仔数存在显著关联,可作为松辽黑猪产仔数的遗传标记,用于分子育种。     

松辽黑猪OAS1基因多态性及其与繁殖性状的关联分析

为探究寡腺苷酸合成酶1（oligoadenylate synthase 1,OAS1）基因多态性与松辽黑猪繁殖性状的关联性,试验选取130头松辽黑猪母猪为研究对象,利用Sanger直接测序法测序查找OAS1基因外显子1~8的SNP位点,使用SPSS 19.0软件分析OAS1基因SNP位点与松辽黑猪繁殖性状的关联性。结果显示,在松辽黑猪OAS1基因外显子2、3和6上共检测到33个突变位点;其中在外显子2的110 bp处存在1个SNP位点（G110C）,存在3种基因型：GG、GC和CC;在外显子3的176 bp处存在1个SNP位点（C176T）,存在3种基因型：CC、CT和TT;在外显子6的145 bp处存在1个SNP位点（C145T）,存在3种基因型：CC、CA和AA;在166 bp处存在1个SNP位点（G166A）,存在3种基因型：GG、GA和AA;在206 bp处存在1个SNP位点（A206G）,存在3种基因型：AA、AG和GG。卡方适合性检验结果显示,松辽黑猪OAS1基因G110C突变位点符合Hardy-Weinberg平衡状态,C176T、C145A、G166A和G206A位点均偏离Hardy-Weinberg平衡状态。群体遗传参数分析结果显示,各SNPs位点遗传杂合度均位于中等水平,为中度多态（0.25<PIC<0.5）。关联分析结果发现,G110C位点GC基因型个体总产仔数、产活仔数和断奶仔猪数均显著高于GG基因型个体（P<0.05）;C176T位点CT基因型个体断奶仔猪数显著高于CC基因型个体（P<0.05）;C145T位点CC基因型个体总产仔数和产活仔数均显著高于AA基因型个体（P<0.05）;G166A位点GA基因型个体断奶仔猪数显著高于GG基因型个体（P<0.05）;A206G位点GG基因型个体总产仔数和产活仔数显著高于AA基因型个体（P<0.05）。结果表明,OAS1基因外显子区存在突变位点,对松辽黑猪部分繁殖性状有显著性影响。     

苯并芘对猪卵母细胞SHH信号通路影响的研究

如今空气污染问题仍广泛存在,其中苯并芘（Benzo （a） pyrene,BaP）是一种常见的空气污染物,对人和动物的健康具有严重影响。据报道,BaP可以显著降低卵母细胞IVM （卵母细胞体外成熟）能力,但机制尚不清楚。因此,本研究在猪卵母细胞IVM培养液中添加不同浓度BaP （50 μM、100 μM、200 μM）检测其对卵母细胞IVM能力的影响,并检测此过程SHH信号通路相关基因和蛋白的表达。结果显示50 μM BaP处理组卵母细胞IVM能力显著低于对照组（76.5 vs 68.1,P<0.05）,并随着BaP浓度增加进一步下降。同时,qPCR和免疫荧光染色检测发现,50 μM BaP处理组卵母细胞中SHH信号通路相关基因mRNA和蛋白表达量,均显著低于对照组（P<0.05）。因此,本研究初步证明BaP可能通过破坏SHH信号通路降低猪卵母细胞IVM能力。     

Green tea polyphenols supplementation alters immunometabolism and oxidative stress in dairy cows with hyperketonemia

Peripartal cows often experience negative energy balance, and are therefore prone to suffering from metabolic diseases such as hyperketonemia, which causes financial losses in dairy farms. This study aimed to investigate the effect of green tea polyphenol (GTP) supplementation during the periparturient period on production performance, oxidative stress and immunometabolism in dairy cows with hyperketonemia. One hundred Holstein cows were assigned to GTP (0.2 g/kg DM; n = 50) or control (without GTP; n = 50) group based on body weight, previous milk yield, and parity on d 15 before expected parturition. Subsequently, 10 cows with hyperketonemia were selected from each group, according to blood β-hydroxybutyric acid (BHBA) concentration between 1.2 and 2.9 mmol/L from 2 to 3 d postpartum. All cows were fed a close-up diet and a lactation diet with or without GTP supply from 15 d prepartum until 30 d postpartum. Milk and blood samples were obtained from 20 cows selected with hyperketonemia on 10, 20, and 30 d postpartum. Compared with control cows, greater milk yield and lower somatic cell count were observed in GTP cows. The GTP group had lower concentrations of BHBA, free fatty acids, cholesterol, triglyceride, reactive oxygen species, malondialdehyde, and hydrogen peroxide, greater concentrations of glucose, lower activities of aspartate aminotransferase, alanine aminotransferase, and glutamyl transpeptidase, alongside greater activities of superoxide dismutase, glutathione peroxidase, and total antioxidant capacity. Additionally, GTP supplementation up-regulated concentrations of interleukin-6 and interleukin-10, but down-regulated concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-2, interleukin-8, and interferon-γ in plasma. Greater concentrations of plasma immunoglobulin G were also detected in the GTP group. Overall, the data suggested that GTP supplementation from 15 d prepartum to 30 d postpartum improved the milk yield and health status in cows with hyperketonemia during early lactation.     

冬小麦种质“矮孟牛”中新型小麦-黑麦复杂易位的遗传传递分析

本文利用矮孟牛中一种新型小麦-黑麦复杂易位IRS·7DS,1BL·7DL与1BL·1RS易位所构建的一套重组自交系(包括109个F6株系)和8个矮孟牛衍生品种进行了分析,结果表明在109个F6株系中,纯合复杂易位占69个,纯合1BL·1RS占36个,二者比例约2∶1,杂合类型占4个,出现频率4/109;对矮孟牛8个衍生品种的分析表明,6个品种包含来自矮孟牛的