农村养鸡防疫存在问题及对策

科学疫病防控是现代化家禽养殖产业发展中一项重要和不可或缺的工作,是实现鸡养殖产业健康可持续发展的关键所在。由于农村地区的很多养殖户科学疫病防控重视程度不高,在养殖管理过程中并没有构建完善的疫病防控措施,养殖管理方案不当,鸡群处于亚健康状态,身体抵抗能力较差,一旦某些传染性疾病在养殖场中爆发流行,很容易造成严重危害,使大量鸡死亡,给养殖场带来不可挽回的经济损失。该文分析农村养鸡防疫存在的问题,论述具体的解决措施。     

Study on Anti-viral Effect of Rhubarb on Porcine Epidemic Diarrhea Virus in vitro

The inhibitory effects of rhubarb on cell lesion, which was caused by porcine epidemic diarrhea virus (PEDV), and its mechanism had been studied in this research.The maximum safe concentration of rhubarb solution applied to Vero cells was determined through morphological observation, and then the PEDV infected Vero cell model in vitro was established, by which the inhibitory effect of rhubarb on virus was studied.The experimental method had also been established based on this cell model.The result showed that the maximum safe concentration was 3.28 mg/mL.Both morphological observation and MTT assay were indicated that rhubarb could not only block the attachment of the virus to Vero cell and inhibit the synthesis and reproduction of the virus, but also could directly inactivate the virus.This research showed that rhubarb had an inhibitory effect on PEDV, and provided a theoretical basis for the prevention and treatment of clinical porcine epidemic diarrhea.     

Identification and Molecular Characteristics Analysis of Porcine Epidemic Diarrhea Virus Variants

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.     

基于S基因序列分析新型猪流行性腹泻病毒的遗传演化

最近，笔者实验室在青藏高原地区发现两种新亚型藏猪源猪流行性腹泻病毒（PEDV），为进一步调查新型PEDV是否在四川腹泻猪群中存在或流行，对实验室2018-2019年保存的116份猪腹泻粪便或肠组织样本进行PEDV的检测及其纤突蛋白基因（spike）分子特征研究。结果表明：腹泻样本的PEDV检出率为42.2%（49/116，95% CI=33.1%~51.8%），并获得了13条完整的S基因序列，全长为4 149~4 170 bp，序列相似性为94.2%~99.9%，其中SWUN-H3-CH-SCYA-2019的S基因与藏猪源新G1亚群PEDV的序列相似性高达97.0%~98.6%。遗传演化研究结果表明13株PEDV S基因划分为G1和G2大群，其中SWUN-H3-CH-SCYA-2019位于藏猪源新G1亚群；SWUN-19-CH-SCZY-2018、SWUN-4-CH-SCXC-2018、SWUN-1-CH-SCNJ-2019和SWUN-3CH-CH-SCZG-2019位于G2亚群中一个独立的分支，且与藏猪源新G2亚群毒株有着较近的亲缘关系。为了进一步研究13株PEDV的演化过程，以贝叶斯进化分析软件包（BEAST）进行分歧时间估算，结果表明SWUN-H3-CH-SCYA-2019的分歧时间约为2012.3年，早于藏猪源新G1亚群其余毒株的最早分歧时间（2015.7年）；SWUN-4-CH-SCXC-2018、SWUN-19-CH-SCZY-2018和SWUN-3CH-CH-SCZG-2019的分歧时间约为2014.2年，早于G2亚群的藏猪源毒株2014.7年，所有藏猪源PEDV的分歧时间均晚于四川毒株。本研究在四川地区首次发现了藏猪源PEDV，并且从毒株的分歧时间推断青藏高原的藏猪源PEDV来源于四川，为新型PEDV分子遗传进化的监测提供了依据。     

基层常见猪传染性腹泻病的临床诊断与防控对策

猪传染性腹泻病是猪的常见多发病。基于受基层单位病原检测技术的条件限制,一旦发生传染性腹泻病,基层兽医通常依据临床症状做出初步诊断与治疗,如果临床经验稍有不足,误诊、误治或延误诊治的现象时有发生。为此梳理出了常见的12种猪传染性腹泻病临床诊断要点与综合防治措施,以防止其重大疾病的发生。     

核桃黑斑病综合防治技术

通过对核桃黑斑病发病侵染规律及原因分析,采取加强水肥土管理,合理修剪,减少密蔽,改善通风透光条件;谨防蛀果害虫;越冬管理;药剂防治的综合措施防治,可有效控制核桃黑斑病的发生危害。     

花椰菜主要性状遗传规律以及性状相关性分析

总结了花椰菜的单球质量、成熟期、株高株展、花球品质等主要性状的遗传规律以及性状之间的相关性,并得出结论:花椰菜主要性状遗传规律以及性状相关性研究对育种研究具有重要指导意义,但必须注重与田间调查相结合,只有将遗传规律与田间调查有机结合才能提高育种效率,提高育种水平。     

滇朴干基腐朽病发生规律及综合防治

对滇朴干基腐朽病的调查、室内保湿培养、分离鉴定结果表明：引起滇朴干基腐朽的病原为伏革菌科隔孢伏革菌属的乳白隔孢伏革菌。该病害的发生随温度的升高、树龄的增长、降水的减少而逐渐严重，在土壤瘠薄、保水能力差的区域内发病严重。药剂防治80d后，用急救回生丹：金雷：水=1：1：50树干注射、包干的相对防效达86％；枝、干、叶喷70％甲基托布津＋杀毒矾700倍液的相对防效为79％；根部浇灌58％的雷多米尔＋杀毒矾800倍液的相对防效为81％。     

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。     

规模化养猪场疾病防控

养猪场以前饲养数量较少时采用传统的综合防控措施,如免疫接种、药物预防和治疗等,在疾病防控方面都起到关键作用且效果明显,但是随着饲养数量增加,只采用上述措施远远不够,应树立创新理念和生物安全措施,才能更有效的防控疾病发生。