本文選題：PRRSV + 遺傳變異　； 參考：《吉林大學(xué)》2016年博士論文

【摘要】：本研究采集河南省及周邊不同地區(qū)疑似PRRS病豬的組織病料進(jìn)行PRRSV分離與鑒定,并對分離毒株進(jìn)行遺傳變異分析,以河南省PRRS的流行狀況調(diào)查結(jié)果為基礎(chǔ),研究PRRSV弱毒疫苗、三仁湯復(fù)方中藥制劑和抗PRRSV免疫血清對PRRS的防治效果,以尋求簡捷、有效、實(shí)用的PRRS防控措施,以提高養(yǎng)豬業(yè)的生產(chǎn)水平。1、收集河南省及湖北部分市區(qū)2006~2014年疑似PRRS發(fā)病豬的病料180余份,用Marc-145細(xì)胞培養(yǎng)進(jìn)行病毒分離與鑒定,分離得到10株P(guān)RRSV,其中河南省共9株,湖北襄樊1株,以美國經(jīng)典株ATCCVR-2332和中國經(jīng)典株CH-1a為參考毒株,將分離株的NSP2和ORF5-7片段分別克隆到p MD-18T載體并進(jìn)行測序。結(jié)果表明PRRSV分離株均為美洲型毒株,且Nsp2基因均已發(fā)生了缺失,GP5基因沒有缺失,只有基因位點(diǎn)突變;其中He N1-06與ATCCVR-2332處于同一個(gè)進(jìn)化分支,其它9株毒株(He N2-06,He N3-06,He N1-07,He N2-07,He N3-07,He N1-08,He N2-08,He N3-08,He N4-08)與CH-1a株處于同一進(jìn)化分支。證實(shí)了河南省PRRSV一直在變異,且以高致病性毒株為主流。2、PRRS母源抗體消長規(guī)律試驗(yàn)結(jié)果表明:仔豬母源抗體水平高低和維持時(shí)間長短與母豬初乳抗體水平呈正相關(guān),母源抗體的S/P在14日齡最高,隨后開始下降,21日齡下降速度加快,38日齡降到最低,平均半衰期12天,由此確定免疫方案:仔豬21日齡實(shí)行首免,45日齡二免,劑量1頭份/頭弱毒疫苗肌肉注射,種豬每年普免3次,劑量2頭份/次弱毒疫苗肌肉注射。3、通過三仁湯復(fù)方中藥制劑作用于Marc145細(xì)胞后,觀察Marc145細(xì)胞的病變率(CPE),研究三仁湯復(fù)方中藥對PRRSV在Marc-145上的吸附阻斷作用,增殖抑制作用以及直接滅活作用。結(jié)果表明:三仁湯復(fù)方中藥制劑對PRRSV有較強(qiáng)的吸附阻斷作用、增殖抑制作用和直接滅活作用,其最小質(zhì)量濃度分別為125g/L、31.25g/L和15.625g/L。4、PRRS臨床防控研究結(jié)果表明:按照上述免疫方案實(shí)施后,種豬配種分娩率由76.6%提高到90.5%,提高了13.9%;保育豬成活率由89.0%提高到98.0%,提高了9.0%;育肥豬成活率由87.0%提高到95.0%,提高了8.0%;三仁湯復(fù)方中藥臨床治療1825頭發(fā)生PRRS保育豬成活率達(dá)到99.28%,治療8994頭發(fā)生PRRS的育肥豬成活率達(dá)到95.73%;臨床對照治療實(shí)驗(yàn)結(jié)果表明:用三仁湯復(fù)方中藥治療1153頭發(fā)生PRRS病豬成活率為97.66%,日增重390g/d,用抗生素控制繼發(fā)感染治療1040頭發(fā)生PRRS病豬成活率為80.48%,日增重310g/d,成活率提高了17.18%,日增重提高了80g/d;用三仁湯治療發(fā)生PRRS種豬群配種分娩率提高了39.8%,新生仔豬健仔率提高了53.2%。用抗PRRSV免疫血清在保育豬發(fā)生PRRS早期按1m L/kg體重劑量肌肉注射,成活率由86.56%提高到96.85%,提高了10.32%。綜上所述,本研究證明,河南省PRRSV的感染與流行十分普遍,毒株已發(fā)生了不同程度的變異,運(yùn)用PRRSV弱毒疫苗、三仁湯復(fù)方中藥和抗PRRSV免疫血清聯(lián)合防治PRRS的發(fā)生,有效減少了母豬流產(chǎn)和仔豬死亡,改善了豬群健康狀況,提高豬場的生產(chǎn)水平。
[Abstract]:In this study, tissue samples of suspected PRRS pigs in Henan Province and its surrounding areas were collected for PRRSV isolation and identification, and genetic variation of the isolates was analyzed. Based on the results of the investigation on the prevalence of PRRS in Henan Province, PRRSv attenuated vaccine was studied. The prevention and control effect of Sanren decoction compound traditional Chinese medicine preparation and anti-PRRSv immune serum on PRRS, in order to seek simple, effective and practical preventive and control measures of PRRS. In order to improve the production level of pig industry, more than 180 samples of suspected PRRS infected pigs from 2006 to 2014 in Henan Province and some urban areas of Hubei Province were collected. The virus was isolated and identified by Marc-145 cell culture, and 10 strains of PRRSVS were isolated and identified, of which 9 strains were isolated from Henan Province. The NSP2 and ORF5-7 fragments of Xiangfan strain were cloned into pMD-18T vector and sequenced using the American classic strain ATCCVR-2332 and the Chinese classical strain CH-1a as reference strains. The results showed that PRRSv isolates were all American strains, and the deletion of GP5 gene was not found in all of the Nsp2 genes, but only the mutation of gene locus, in which he N1-06 was in the same evolutionary branch as ATCCVR-2332. The other 9 strains, he N2-06HN3-06H1-06H1-06HN1-07H1-07HN1-07H1-07HN1-08H1-08H3-08He N4-08) are in the same evolutionary branch as the CH-1a strain, and the other 9 strains are in the same phylogenetic branch as the CH-1a strain, and the other 9 strains are in the same phylogenetic branch as the CH-1a strain. It was confirmed that PRRSv had been mutating all the time in Henan Province, and the results showed that the maternal antibody level of piglets and the duration of maintenance were positively correlated with the antibody level of porcine colostrum. The S / P of maternal antibody was highest at 14 days of age, then began to decline, and then began to decline at 21 days of age. The decline rate was accelerated to the lowest at 38 days of age, with an average half-life of 12 days. Thus, the immunization scheme was determined: the first day of 21 days old piglets were given the second immunity at 45 days old. A dose of 1 head / head attenuated vaccine was injected intramuscularly, 3 times a year for pig immunizations, and 2 doses per second for intramuscular injection of attenuated vaccine. The cells were treated with Sanren decoction compound Chinese medicine preparation. To observe the cytopathic rate of Marc145 cells and to study the inhibition of PRRSv on Marc-145, the inhibition of proliferation and the direct inactivation of PRRSv by Sanren decoction. The results showed that Sanren decoction had strong adsorption blocking, proliferation inhibition and direct inactivation on PRRSV, and its minimum mass concentration was 125g / L 31.25g / L and 15.625g / L / L respectively. The birth rate of breeding pigs increased from 76.6% to 90.5%, which increased 13.9%; the survival rate of preserved pigs increased from 89.0% to 98.0, which increased 9.0%; the survival rate of fattening pigs increased from 87.0% to 95.0%, which increased 8.0%. The survival rate of the fattened pigs treated with 8994 hair PRRS was 95.730.The results of clinical controlled treatment showed that the survival rate of 1153 hair pigs treated with Sanren decoction was 97.66, the daily gain was 390g / d, and the secondary infection was controlled by antibiotics. The survival rate was 80.48g / d, the daily gain was 310g / d, the survival rate was 17.18g / d, the daily gain was increased 80g / d, the rate of breeding and delivery was increased by 39.8g / d, and the healthy rate of newborn piglets was increased 53.2d. After intramuscular injection of PRRS with anti-PRRSV immune serum, the survival rate was increased from 86.56% to 96.85 in the early stage of PRRS. In conclusion, this study proved that the infection and prevalence of PRRSv in Henan Province is very common, the virus strains have been mutated to different degrees, using PRRSv attenuated vaccine, Sanren decoction compound Chinese medicine and anti-PRRSV immunized serum to prevent and treat PRRS. It can effectively reduce abortion and piglet death, improve the health status of pig herd and improve the production level of pig farm.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S858.28

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本文編號(hào)：1995980