猪源致病性沙门氏菌耐药基因的分析

采用平板稀释法，选用氨基糖苷类、四环素类、磺胺类和氯霉素类4大类抗生素的11种药物，对30株猪源致病性沙门氏菌进行了药敏试验，结果有28株菌（93．3％）至少对一种药物有耐药性；对四环素、强力霉素、磺胺甲基异口恶唑、复方新诺明、链霉素、卡那霉索和氯霉素有耐药性的菌株较普遍，在所有菌株中占比例分别为83．3％、80％、80％、76．7％、60％、56．7％和56．7％。设计了25对引物，对耐药基因进行了扩增及序列测定，结果扩增到13种耐药基因，与GenBank中的相应基因有很高的同源性（≥98．1％）。30株猪源致病性沙门氏菌中至少含有一种耐药基因的菌株有28株（93．3％），sul1、aph（3′）-Ⅱa、tetC、Catl、tetA和aadAl耐药基因较为普遍，检出率分别为76．7％、60％、60％、43．3％、40％和36．7％。药敏试验结果与耐药基因检测结果有很高的一致性（≥88％）。     

奶牛乳房炎金黄色葡萄球菌8种毒力基因的PCR检测

为研究奶牛乳房炎金黄色葡萄球菌(Staphylococcus aureus,SA)主要毒力因子的分布情况,本试验利用PCR对70株临床型乳房炎、55株隐性乳房炎金黄色葡萄球菌的8种主要毒力基因进行检测。结果显示,nuc、ClfA、TSST-1、PVL、Hla、Hlb、FnBPA和FnBPB 8种毒力基因在临床型乳房炎SA菌株中的检出率分别为:100.0%(70株)、100.0%(70株)、0(0株)、5.7%(4株)、100.0%(70株)、11.4%(8株)、97.1%(68株)和100.0%(70株);在隐性乳房炎SA菌株中的检出率分别为:100.0%(55株)、100.0%(55株)、0(0株)、70.9%(39株)、98.2%(54株)、9.1%(5株)、100.0%(55株)和100.0%(55株)。结果表明,nuc、ClfA、Hla、Hlb和FnBPA 5种毒力基因是引起奶牛乳房炎的SA的最主要毒力基因;PVL基因可能是引起隐性乳房炎的重要致病基因。     

基于RNA-Seq技术筛选影响猪肌纤维性状的候选基因

旨在探究影响肌纤维性状的候选基因和信号通路。本研究以马身猪和大白猪为研究对象,采用HE染色法分析6月龄马身猪和大白猪背最长肌肌纤维直径和密度,利用RNA-Seq分析马身猪和大白猪背最长肌组织中基因表达情况,并对差异表达基因进行GO和KEGG富集分析,再通过qRT-PCR验证RNA-Seq结果的准确性。结果显示,马身猪背最长肌肌纤维直径极显著低于大白猪(P<0.01),而肌纤维密度极显著高于大白猪(P<0.01)。转录组测序结果显示,在6月龄马身猪和大白猪背最长肌中,差异倍数在2倍以上的基因共105个,其中,马身猪相对于大白猪上调的基因有55个,下调的基因有50个。GO富集分析表明,差异表达基因主要富集在与线粒体相关的细胞组分和骨骼肌分化有关的生物学过程中;KEGG分析结果显示,这些差异表达基因主要参与到与氧化磷酸化有关的信号途径。qRT-PCR和RNA-Seq对6个DEGs表达情况的检测结果表明它们的表达趋势相同,说明RNA-Seq结果准确可靠。结合差异基因表达丰度、GO和KEGG富集分析结果,本研究发现,MYL3、MYH3、MYH6基因通过影响肌纤维的组成而影响肌纤维特性,ND6基因通过参与线粒体氧化磷酸化过程影响肌纤维类型,MICU2基因通过维持线粒体Ca²⁺浓度的稳态影响细胞功能,编码转录因子的EGR1和FOS基因通过调节细胞增殖、分化和凋亡等过程影响肌肉生长发育。本研究初步揭示了造成马身猪和大白猪肌纤维性状差异的主要原因,为猪肉品质的改善提供了相应的理论依据。     

四川冬春季参考作物蒸散量时空变化及其成因

为深入认识四川冬春季参考作物蒸散量(ET₀)的变化特征,利用1980—2016年四川35个气象站的逐日气象观测资料,采用泰森多边形、气候倾向率和克里金空间插值等方法对其冬春季ET₀的时空变化特征进行分析,并通过敏感性和贡献率分析了ET₀的变化成因。结果表明:ET₀的年代际变化呈先降后增的趋势,空间上呈明显的西南高东部低的分布特征,且高值区范围持续扩大,低值区范围波动缩小。ET₀的年际变化呈上升趋势,春季ET₀气候倾向率和空间差异明显大于冬季,且ET₀高值区与低值区空间分布受海拔高度影响明显。ET₀的同一日多年平均值自初冬至初春逐渐上升,1月22日—5月2日仅有8 d的ET₀值低于多年日平均值,具有明显连续的高值时段。ET₀对日照时数的变化最敏感,其次是对相对湿度与平均气温,对三者均呈高敏感性。平均气温的正贡献率是引起ET₀变化的主导因子,其次是相对...     

Research Progress on Bacterial Resistance in Environment

Antibiotic resistance genes have become a recognized environmental pollutant, threatening the ecological safety and human health. The application of antibiotics in the clinical and animals breeding, making the environmental microorganisms living with the impact of the residue of antibiotics and elements of resistance genetic and leading to antibiotic resistant bacteria to gain a competitive advantage and destroyed the stability of ecosystem. In this paper, we expounded the concept of resistance gene transmission by the view of macro environment. Through analyzing the mechanism of resistance development,environmental pollution caused by drug resistance and the impact of environmental microorganism for drug resistance, we clarified the key role of the environment in the development of the characteristics of bacterial resistance and analysis environmental resistance.Such as the ecological diversity of flora, the types and the spread of resistant bacteria, the residues of antibiotics and the transmission of the resistance genes.     

新疆与美国早实核桃(Juglans regia L.)品种生长结实与成花特性比较

以引进的美国核桃品种‘维纳’、‘艾米格’和新疆本地品种‘温185’和‘新新2号’为试材,研究分析不同品种的植物学和生物学特性、花芽分化过程以及相关成花基因的表达。结果显示:4个品种在树体生长、枝芽发育特性、物侯期以及成花率和坐果率方面没有明显差异。在结果枝同一坐果部位形成双果、三果或以上的能力‘新新2号’明显强于‘维纳’,但‘新新2号’的果实大小和单果重都低于‘维纳’,而‘维纳’平均结果母枝上形成侧花芽的数量和坐果率高于‘新新2号’。从萌芽到开花结束直至当年侧芽发育过程中‘维纳’混合芽中的JrFT和JrLFY基因表达也高于‘新新2号’,而同时期的JrFLC基因表达则相对较低,说明‘维纳’在花芽分化和花器官形成过程中通过高表达JrFT和JrLFY基因,低表达JrFLC基因促进结果母枝上形成大量侧花芽,并利用大量侧花芽抽生结果枝坐果,形成较大的果实,使得最终产量并不低于同一个坐果部位大量着生2个或多个果实的‘新新2号’品种。     

Analysis on Target Genes with Mutation in microRNA of Blue-shelled Chicken and White Leghorn

The study was conducted to explore the potential different characters between Blue-shelled chicken and White leghorn.Global genome microRNA was combined the identified microRNA with complementary lab-predicted microRNA.Then the two breed chicken's SNP data got by GGRS were mapped to the microRNA and focused on SNP that deliberately located in mature-microRNA.Bioinformatics method was adopted for target prediction on microRNA which had SNPs.By further gene enrichment analysis,the study found these genes enriched in 22 GO terms,10 KEGG pathways,and 3 IPA important networks.And they enriched in traits which associated with growth,such as mTOR signaling pathways,Wnt signaling pathways,growth hormone receptor networks and insulin-like growth factor Ⅰ receptor networks.And they also enriched in some laying traits,such as oocyte meiotic signaling pathways and progesterone mature oocytes signaling pathways.The methods and the results might provide references for further studies.     

Genetic potential for residual feed intake and diet fed during early- to mid-gestation influences post-natal DNA methylation of imprinted genes in muscle and liver tissues in beef cattle

Discovery of epigenetic modifications associated with feed efficiency or other economically important traits would increase our understanding of the molecular mechanisms underlying these traits. In combination with known genetic markers, this would provide opportunity to improve genomic selection accuracy in cattle breeding programs. It would also allow cattle to be managed to improve favorable gene expression. The objective of this study was to identify variation in DNA methylation between beef cattle of differential pre-natal nutrition and divergent genetic potential for residual feed intake (RFI). Purebred Angus offspring with the genetic potential for either high (HRFI) or low (LRFI) RFI were prenatally exposed to either a restricted maternal diet of 0.5 kg/d average daily gain (ADG) or a moderate maternal diet of 0.7 kg/d ADG from 30 to 150 d of gestation. We performed DNA methylation analysis of differentially methylated regions (DMR) of imprinted genes (Insulin-like growth factor 2 (IGF2) DMR2, IGF2/H19 imprinting control region (ICR) and IGF2 receptor (IGF2R) DMR2) using post-natal samples of longissimus dorsi (LD) muscle taken from male and female calves at birth and weaning, and of LD muscle, semimembranosus (SM) muscle, and liver samples collected from steers at slaughter (17 months of age). Interestingly, for all three DMR investigated in liver, LRFI steers had higher levels of methylation than HRFI steers. In LD muscle, IGF2/H19 ICR methylation differences for heifers at birth were due to pre-natal diet, while for steers at birth they were mostly the result of genetic potential for RFI with LRFI steers again having higher levels of methylation than HRFI steers. While results from repeated measures analysis of DNA methylation in steers grouped by RFI revealed few differences, in steers grouped by diet, we found higher methylation levels of IGF2 DMR2 and IGF2R DMR2 in LD muscle of restricted diet steers at weaning and slaughter than at birth, as well as increased methylation in LD muscle of restricted diet steers compared with moderate diet steers at weaning and/or slaughter. Our results suggest that differential pre-natal nutrition, and divergent genetic potential for RFI, induces tissue- and sex-specific alterations in post-natal IGF2 and IGF2R methylation patterns and that these patterns can vary with age in Angus beef cattle.     

NPFFR1基因对鹅卵泡颗粒细胞激素分泌和细胞凋亡的影响研究

神经相关肽受体（RFamide-related peptide receptor,NPFFR1）是促性腺激素抑制激素的主要亲和受体,它在调控动物繁殖方面起着重要作用。为了解NPFFR1对鹅卵巢卵泡发育的作用,本研究以42周龄健康产蛋四川白鹅为试验材料（n=9）,利用RT-qPCR法检测NPFFR1基因在等级前和等级卵泡颗粒细胞中的mRNA表达规律;在颗粒细胞中过表达NPFFR1基因,酶联免疫吸附法检测颗粒细胞上清液（n=9）中雌二醇（estradiol,E2）、孕酮（progesterone,P4）和抗缪勒管激素（anti-Mullerian hormone,AMH）的浓度变化,剩余贴壁细胞作一步法TUNEL检测细胞凋亡情况;转录组测序方法筛选大黄卵泡（8~10 mm）颗粒细胞过表达NPFFR1前后表达差异显著基因,并对差异表达基因进行功能聚类分析。结果显示,除F1等级外,其余等级卵泡颗粒细胞NPFFR1表达量均极显著高于等级前卵泡（P<0.01）;过表达NPFFR1后,等级颗粒细胞上清液中的E2和等级前颗粒细胞上清液AMH的含量显著（P<0.05）降低,但孕酮P4含量变化不显著（P>0.05）;转录组测序共筛选到267个差异表达基因（119个下调,148个上调）,这些基因主要富集在生物节律过程、繁殖进程等生物学过程中;同时,与对照组相比,差异基因AMH显著下调表达（P<0.05）,Clock（clock circadian regulator）、FOS（proto-oncogene,AP-1 trans-cription factor subunit）、Per（period circadian regulator）和ANTXR2（cell adhesion molecule 2）分别极显著（P<0.01）或显著（P<0.05）上调表达。上述试验结果提示,NPFFR1可从激素、细胞凋亡和生物节律等多个环节影响卵泡颗粒细胞,参与调控卵泡的时序等级发育。     

广东省养禽场肠球菌耐药性及毒力因子流行分布特征

旨在调查和分析广东省养禽场肠球菌的亚型屎肠球菌和粪肠球菌耐药性及其毒力因子流行分布特征,为控制禽源肠球菌耐药性传播、保障公共卫生安全提供理论依据。作者于2018年从广东省4个养禽场采集肠道样品493份,进行屎肠球菌和粪肠球菌的分离鉴定;采用琼脂二倍稀释法测定肠球菌的最小抑菌浓度（MIC）;PCR方法检测肠球菌的耐药基因和毒力基因。结果显示：1）共分离到125株肠球菌,其中粪肠球菌84株（鸡源66株,鸭源18株）;屎肠球菌41株,均来自鸡肠道样本。2）菌株对四环素、多西环素、红霉素几乎全部耐药,对氟苯尼考和氯霉素的耐药率高达89.60%和74.40%。屎肠球菌耐药率普遍高于粪肠球菌,而粪肠球菌对环丙沙星和利奈唑胺的耐药率高于屎肠球菌;鸭源粪肠球菌对利奈唑胺的耐药率（94%）显著高于鸡源粪肠球菌（39.4%）,屎肠球菌对利奈唑胺均敏感。从鸡分离的1株粪肠球菌对万古霉素耐药。3）耐药基因在屎肠球菌中的检出率高于粪肠球菌,鸭源分离株检出率高于鸡源。耐药基因tetL、fexA、ermB最为流行,检出率均高于90%。其次是optrA基因,检出率为73.60%,poxtA和fexB的检出率均低于20%。在3株鸭源粪肠球菌中检测出cfr基因。4）已检测的毒力基因中efaA的携带率最高,为63.04%（58/92）,其他依次为gelE（54.35%,50/92）、ace（47.83%,44/92）、asa1（44.57%,41/92）。对环丙沙星及高浓度氨基糖苷类耐药的菌株及携带cfr基因的菌株,大多携带agg、asal、gelE或ace。本研究显示养殖场禽源肠球菌耐药严重,鸭源肠球菌对利奈唑胺耐药率高,耐药基因和毒力基因流行且多样,且检测出人医临床重要抗生素耐药基因,应加强对养禽场肠球菌耐药性监测。