表达猪生长激素基因的重组腺病毒的构建及对猪生长的促进效果观察

利用复制缺陷型重组腺病毒作为基因表达载体,将猪生长激素（pGH）基因直接转导到猪体内细胞,研究猪生长激素基因在猪体内的促生长作用.用同源重组的方法,构建含pGH基因的重组腺病毒（Ad-pGH）.将其感染293细胞和PK-15细胞,均可检测到pGH.用1 mL此重组腺病毒（1010TCID50）一次性肌肉注射60日龄的杂交长白猪.结果表明,试验组的猪平均增重比对照组的猪增加37%（注射后第4周）和19%（注射后第6周）;饲料报酬提高28%（注射后第4周）和18%（注射后第6周）.试验结果证明,由重组腺病毒表达的pGH具有生物学活性,在猪体内起到明显的促进生长的作用.重组腺病毒介导的pGH基因在猪体内的表达可维持约4周.     

2019—2020年新疆部分地区猪PCV2的抗体水平检测及分析

为了解2019—2020年新疆地区猪圆环病毒病2型（PCV2）抗体水平及其消长规律,本试验采用间接酶联免疫吸附试验（ELISA）方法对475份血清中PCV2免疫抗体水平进行检测和分析。结果显示,PCV2抗体平均阳性率为69.68%（331/475）,未达到国家规定标准（70%）。S/P的平均值为1.56。不同类别猪群抗体平均阳性率在42.50%~86.67%之间,有一定的差异。在调查的5个规模化养殖场中,仅有2个场PCV2免疫抗体阳性率达到国家标准。各场应根据抗体检测结果,进一步对现有的免疫程序进行调整和完整,确保各猪群健康。     

AR regulates porcine immature Sertoli cell growth via binding to RNF4 and miR-124a

Sertoli cells are the only somatic cells in the seminiferous epithelium which directly contact with germ cells. Sertoli cells exhibit polarized alignment at the basal membrane of seminiferous tubules to maintain the microenvironment for growth and development of germ cells, and therefore play a crucial role in spermatogenesis. Androgens exert their action through androgen receptor (AR) and AR signalling in the testis is essential for maintenance of spermatogonial numbers, blood–testis barrier integrity, completion of meiosis, adhesion of spermatids and spermiation. In the present study, we demonstrated that AR gene could promote the proliferation of immature porcine Sertoli cells (ST cells) and the cell cycle procession, and accelerate the transition from G1 phase into S phase in ST cells. Meanwhile, miR-124a could affect the proliferation and cell cycle procession of ST cells by targeting 3′-UTR of AR gene. Furthermore, AR bound to the RNF4 via AR DNA-binding domain (DBD) and we verified that RNF4 was necessary for AR to regulate the growth of ST cells. Above all, this study suggests that AR regulates ST cell growth via binding to RNF4 and miR-124a, which may help us to further understand the function of AR in spermatogenesis.     

免疫小鼠血清中猪圆环病毒2型抗体ELISA检测试剂盒的研究

为研制一种小鼠血清中猪圆环病毒2型（Porcine circovirus type 2,PCV2）抗体检测试剂盒,以PCV2 Cap蛋白兔多抗作为包被用抗体,捕获纯化的PCV2 Cap蛋白,制备抗原检测板,辣根过氧化物酶(HRP)标记羊抗小鼠IgG作为二抗,建立检测小鼠血清中PCV2抗体ELISA检测方法并制备试剂盒。采用所制备的ELISA试剂盒对50份已知阴性血清样本进行检测,临界OD450nm值为0335;采用ELISA试剂盒与IFA分别对100份血清检测,总符合率为96%,检测小鼠血清敏感性效价达1∶16000,与其他常见的猪病毒小鼠阳性血清无交叉反应,2～8 ℃保存15个月稳定,批内变异系数为2.28%～4.17%,批间变异系数为4.27%～6.02%,具有良好的敏感性、特异性、稳定性及重复性;采用ELISA试剂盒对不同来源疫苗免疫组小鼠血清进行检测,均显示良好效果。研究表明,本研究制备的ELISA试剂盒可用于小鼠血清中PCV2抗体效价检测。     

安徽部分地区冬春季仔猪腹泻的流行病学调查与分析

2012年1-3月间,通过定点监测、问卷调查、实地调查和实验室检测等方式,对安徽省16个市、50个县的757个养猪场(户)的仔猪腹泻发病情况进行了流行病学调查,并采集74份发病或病死仔猪样品肠内容物进行实验室检测.结果表明,安徽省仔猪腹泻主要以流行性腹泻、猪传染性胃肠炎为主,其中猪流行性腹泻病毒阳性率为89.2％,猪传染性胃肠炎病毒样阳性率为10.8％,饲养管理条件差和环境气候多变等是冬春季仔猪腹泻多发的重要诱因.     

猪轮状病毒重组VP7蛋白抗原间接ELISA诊断方法的建立

本研究以纯化的原核表达的猪轮状病毒VP7抗原表位区域为抗原,建立了检测猪轮状病毒抗体的间接ELISA诊断方法。特异性试验表明,该抗原与其他7种常见猪病病毒（TGEV、PEDV、CSFV、PCV2、PRRSV、PPV、PrV）的阳性血清不发生交叉反应,批内和批间重复性试验的变异系数均小于10％;对来自不同猪场的血清的检测结果表明,该ELISA方法与中和试验检测结果符合率达94．8％。本试验建立的ELISA诊断方法具有良好的重复性、敏感性和特异性,为PRV的快速诊断、免疫猪群抗体监测和轮状病毒流行病学调查提供了一种快速、简便的血清学诊断方法。     

Analysis of the risk of introduction and spread of porcine reproductive and respiratory syndrome virus through importation of raw pigmeat into New Zealand

AIMS: To determine the frequency with which porcine reproductive and respiratory syndrome (PRRS) virus (PRRSv) would become established in a non-commercial pig herd in New Zealand due to illegal feeding of uncooked food waste containing virus-contaminated pigmeat. To determine the likelihood of a single incursion resulting in a multi-farm outbreak of the disease, and describe the spatio-temporal characteristics of such an outbreak.

METHODS: A Monte Carlo simulation model was constructed to determine the expected annual frequency of PRRSv infection being initiated in a non-commercial pig herd as a result of inadvertent feeding of pigmeat imported from countries endemically infected with the disease. Once the likelihood of PRRSv becoming established in a single pig herd was determined, stochastic spatially explicit infectious disease modelling software was utilised to model the temporal and spatial characteristics of the resulting epidemic.

RESULTS: Assuming the proportion of imported pigmeat remained at current levels, consumption patterns of pigmeat in households in New Zealand remained steady, and limited compliance with recently reintroduced regulations to prevent feeding of uncooked food waste, at least 4.3 pig herds per year were predicted to become infected with PRRSv. Simulation modelling of PRRSv epidemics related to initial infection of a non-commercial farm produced an estimate that 36% of these incursions would spread from the initial herd, and that these outbreaks would involve 93 herds on average in the first year. By increasing the estimated persistence of PRRSv infection in small herds, an average of 205 herds became infected in the first year.

CONCLUSIONS: Given a mean of 4.3 infected premises per year and a 36% probability of infection spreading beyond the initial infected herd, there was a 95% likelihood of a multi-farm PRRS outbreak occurring within 3 years.

CLINICAL RELEVANCE: Introduction of PRRSv through importation of virus-contaminated pigmeat presents a high risk for establishment of the disease in the pig industry in New Zealand.     

福建莆田地区猪圆环病毒病的血清学调查

为了解莆田地区猪圆环病毒病的流行状况,2013—2014年间在莆田笏石、埭头、灵川、仙游地区等部分养猪场(户)共采集了322份未经PCV2疫苗免疫的猪血清样本,采用ELISA(酶联免疫吸附试验)方法对送检血清样品进行PCV2抗体水平检测。结果表明,322份被检血清中检出117份阳性血清,总阳性率为36.33%,其中以6~12周龄的小猪感染率最高,阳性率为55.56%。说明猪圆环病毒病在莆田市多数猪场中存在。     

猪捷申病毒TaqMan实时定量RT-PCR方法的建立

为建立猪捷申病毒(PTV)的早期检测及定量分析方法,本研究基于PTV 11个血清型基因组5′端非编码区保守序列,设计引物和TaqMan探针,建立了检测PTV的TaqMan实时定量RT-PCR方法.应用该方法对PTV、猪细小病毒、猪繁殖与呼吸综合症病毒、猪圆环病毒2型、猪伪狂犬病毒以及猪瘟病毒进行特异性试验,结果除PTV为阳性外其它均为阴性;针对PTV最低可检测到10个拷贝;批内、批间重复试验的变异系数均小于3％.应用建立的方法与病毒分离方法分别对91份临床样品进行检测,检出率分别为79.12％和57.14％,两者的符合率是78.02％.经临床应用表明,该实时定量RT-PCR方法可为PTV的早期诊断及定量分析提供技术手段.     

贵州省猪传染性胃肠炎及呼吸冠状病毒病血清学调查

用酶联免疫吸附试验对采自贵州省内88个县(市)的2906份血清进行了猪传染性胃肠炎病毒及呼吸冠状病毒抗体检测,结果检出猪传染性胃肠炎病毒抗体阳性血清12份,总体阳性率为0.41%;猪呼吸冠状病毒抗体阳性血清44份,总体阳性率为1.51%;2818份血清两种抗体均呈阴性(阴性率为96.97%),32份血清检测无效或无结论(无效率为1.10%)。