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检测猪圆环病毒2型血清中和抗体阻断ELISA方法的建立及应用

检测猪圆环病毒2型血清中和抗体阻断ELISA方法的建立及应用

引用

中国畜牧兽医学会家畜传染病学分会第七届全国会员代表大会暨第十三次学术研讨会

作者：黄立平刘长明危艳武陆月华郭龙军温士刚武嘉男吴洪丽中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室

采用杂交瘤细胞融合技术,获得了一株具有分泌抗猪圆环病毒2型(PCV2)中和活性的单克隆抗体,利用该株单抗建立了一种阻断ELISA方法。用于检测该病毒感染猪血清中和抗体.该方法先用病毒多克隆抗体包被ELISA板以捕获已知病毒抗原,然后与待... 详细信息

采用杂交瘤细胞融合技术,获得了一株具有分泌抗猪圆环病毒2型(PCV2)中和活性的单克隆抗体,利用该株单抗建立了一种阻断ELISA方法。用于检测该病毒感染猪血清中和抗体.该方法先用病毒多克隆抗体包被ELISA板以捕获已知病毒抗原,然后与待检猪血清抗体及辣根过氧化物酶标记的单抗进行反应,通过测定酶标单抗显示阻断ELISA检测结果。该方法与常规免疫过氧化物酶单层细胞试验(IPMA)对353份试验猪血清样品进行了平行试验,两种方法检测符合率为96.0%;该方法的敏感性为97.9%,特异性为92.4%,与其它几种猪病毒参考阳性血清无交叉反应.用阻断ELISA和血清中和试验检测PCV2血清中和抗体结果表明,两种方法呈显著正相关性(r=0.9970),其敏感性优于血清中和试验。用阻断ELISA对PCV2灭活疫苗免疫与攻毒试验猪血清样品检测结果显示,疫苗接种2 w开始能够检测到病毒中和抗体,4w后血清抗体阳转率达100%;此外,对来自东北3省不同地区送检的703份血清样品进行检测,抗体阳性检出率为73.4%,表明我国养猪场中PCV2感染率较高,该病毒危害严重.本研究建立的阻断ELISA具有操作简便、特异和敏感等特点,可为PCV2流行病学调查及血清抗体检测提供了一种有效技术手段.

关键词：猪圆环病毒2型中和抗体检测单克隆抗体阻断ELISA

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Protection of Selenium on Hepatic Mitochondrial Respiratory Control Ratio and Respiratory Chain Complexes Activities in Ducklings Intoxicated with Aflatoxin B1

Protection of Selenium on Hepatic Mitochondrial Respiratory ...

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中国畜牧兽医学会家畜内科学分会第七届代表大会暨学术研讨会

作者： Dayou Shi~(a1).Shining Guo~(a1).Shenquan Liao~(b1).Rongsheng Su~a Mingsheng Guo~a.Nian Liu~a.Pengfei Li~a.Jiqiang Pan.Zhaoxin Tang~(a*) (aCollege of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China b Institute of Veterinary Medicine,Guangdong Academy of Agricultural Sciences,Guangzhou,510640, China)

To investigate the protection of selenium on hepatic mitochondrial functions,90 seven-day-old ducklings were randomly divided into three groups（groupsⅠ-Ⅲ）.groupⅠwas used as a blank control. groupⅡwas administered with aflatoxin B（0.1 mg/kg body weight）.groupⅢwas administered with afiatoxin B（0.1 mg/kg body weight） plus selenium（sodium selenite,1 mg/kg body weight）.All treatments were given once daily for 21 *** results showed that the activities of hepatic mitochondrial complexesⅠ-Ⅳin groupⅡducklings significantly decreased when compared with groupⅠ（P<0.01）.Furthermore,the activities of hepatic mitochondrial complexesⅠ-Ⅳin groupⅢsignificantly increased when compared with groupⅡ（P<0.05）.The hepatic mitochondrial respiratory control ratio （RCR） in groupⅡducklings significantly decreased when compared with groupⅠ（P<0.01）.In addition,the hepatic mitochondrial RCR in groupⅢsignificantly increased when compared with groupⅡ（P<0.05）. These results revealed that the aflatoxin B significantly induced hepatic mitochondrial dysfunction in the activities of hepatic mitochondrial respiratory chain complexesⅠ-Ⅳand the RCR in ***, sodium selenite could significantly ameliorate the negative effect induced by aflatoxin B.

关键词： Sodium selenite Hepatic Mitochondrial Respiratory chain complexes Respiratory control ratio Aflatoxin B1

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Genome shuffling of Megasphaera elsdenii for improving acid-tolerance and propionate production

Genome shuffling of Megasphaera elsdenii for improving acid-...

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中国畜牧兽医学会家畜内科学分会第七届代表大会暨学术研讨会

作者： Long Miao~(1,2),Li Peng~(1,2),Zhang Yi~(1,2),Wang ZHe~2,Liu Guowen~(2*) (1 College of Animal Science and Veterinary Medicine,Shenyang Agricultural University,120 Dongling Road,Shenyang 110866,PR China 2 College of Animal Science and Veterinary Medicine,Jilin University,5333 Xi'an Road Changchun 130062,PR China)

The technology of genome shuffiing has been presented as a novel whole genome engineering approach for the rapid improvement of cellular phenotypes without knowing its genetic *** this study,we used genome shuffiing in an attempt to improve propionate production and acid-tolerance of the wild-type strain *** *** initial mutant population of M. elsdenii H6 were treated with UV irradiation and NTG,resulting in an improved population producing more propionate than the original strain at pH *** mutants were used as the starting strains for 3 rounds of genome *** each round,the pH of the medium plates used for selection was decreased by *** improved strains were selected based on their acid tolerance and propionate *** libraries were established after each round and these improved mutant served as the starting pool for the next round of *** acid tolerance of the mutants on the gradient plate increased progressively with each round of shuffiing up to 3 *** mutants after three rounds of genome shuffiing exhibited an improved acid tolerance and propionate production,which could grow at pH 5.0 and the yield of propionate was increased 41.5±1.4%compared with the original strain after fermented for *** mutant F32 of *** H6 with the characterization of the high propionate production and acid tolerance could be potential candidates for preventing rumen acidosis and production disease during the transition period of dairy cows.

关键词： genome shuffling M.elsdenii acid-tolerance propionate

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Involvement of matrix metalloproteinase-2 in the medial hypertrophy of pulmonary arterioles in broilers with pulmonary arterial hypertension

Involvement of matrix metalloproteinase-2 in the medial hype...

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中国畜牧兽医学会家畜内科学分会第七届代表大会暨学术研讨会

作者： Xun Tan~*,Shi-Cheng Bi,Wen-Wen Li,Jun-Jun Li (Key Laboratory of Animal Virology of Ministry of Agriculture,College of Animal Sciences, Zhejiang University,Hangzhou,China)

Medial hypertrophy of pulmonary arterioles during pulmonary arterial hypertension(PAH) in humans is associated with enhanced proliferation of smooth muscle cells(SMCs).Elevated matrix metalloproteinase (MMP)-2 has been found in pulmonary artery SMCs(PA-SMCs) in patients with idiopathic PAH,leading to the hypothesis that MMP-2 contributes to the proliferation and migration of SMCs in the progression of *** models of PAH are thus needed to further investigate the role of MMP-2 in human *** growing meat-type chickens(broilers) provide a model of spontaneous PAH caused by excessive pulmonary blood flow. The present study was conducted to determine whether the MMP-2 is involved in PA-SMCs proliferation and the medial hypertrophy in this model. Cultured PA-SMCs from normal birds were used to evaluate the effect of MMPs on cell *** zymography showed that the endothelin(ET)-l-induced proliferation of PA-SMCs was concomitant with increased pro and active MMP-2 *** transcription PCR demonstrated an up-regulation of MMP-2 mRNA. However,PA-SMC proliferation was inhibited by MMP inhibitors doxycycline and *** vivo experiments revealed a significant increase of MMP-2 expression(P < 0.01) in the hypertrophied pulmonary arterioles of PAH broilers,which was positively correlated to the wall thickness and medial *** context,this study suggests that the MMP-2 might contribute to the medial hypertrophy in pulmonary arterioles during PAH by enhancing the proliferation of vascular smooth muscle cells. This model of PAH may be valuable for researches focused on targeting the MMPs to prevent and reverse human PAH.

关键词： Pulmonary arterial hypertension Matrix metalloproteinase-2 Smooth muscle cell Broiler

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Effects of selenium form on blood and milk selenium concentrations,milk component and milk fatty acid composition in dairy cows

Effects of selenium form on blood and milk selenium concentr...

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中国畜牧兽医学会家畜内科学分会第七届代表大会暨学术研讨会

作者： Linwu Ran,Xianshi Wu,Xiangzhen Shen,Kechun Zhang,Fei Ren and Kehe Huang a Institute of Nutritional and Metabolic Disorders in Domestic Animals and Fowls, Nanjing Agricultural University,Nanjing 210095,China b College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471003,China c Institute of Dairy Research in Shanghai,Shanghai 200032,China

BACKGROUND:Human health may be improved if milk with a favorable fatty acid composition and Se concentration is ingested. The present study is to determine howa basal diet supplemented with daily 5 mg Se as Se-enriched yeast(SY) or sodium seienite (SS) affects the fatty acid composition and Se concentration of bovine *** effects of Se form on blood Se concentration, erythrocyte glutathione peroxidase 1(GPxl) activity,serum GPx3 activity and milk yield and component were also studied. RESULTS:Both Se forms,when compared to control group,increased Se concentrations of blood(P < 0.01) and milk(P < 0.01), erythrocyte GPx1 activity(P < 0.05) and milk percentages of polyunsaturated fatty acids(PUFA)(P < 0.05) and c/'s-9,cis-12 linoleicacid(P < 0.05).Cows supplemented with SY had higher Se levels in blood(P < 0.01) and milk(P < 0.01) and percentage of PUFA in milk(P < 0.05) when compared with those supplemented with *** yield,milk component and serum GPx3 activity were not significantly affected by Se form. CONCLUSION:Supplementation of diet with SY appears to be of more benefit than SS in producing favorable milk with high PUFA and Se concentrations.

关键词： selenium form milk Se fatty acid composition dairy cow

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Effects of Zearalenone on IL-2,IL-6,and IFN-γ mRNA levels in the splenic lymphocytes of chickens

Effects of Zearalenone on IL-2,IL-6,and IFN-γ mRNA levels i...

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中国畜牧兽医学会家畜内科学分会第七届代表大会暨学术研讨会

作者： Y.C.Wang~1,J.L.Deng~(1,*),S.W.Xu~(2*),X.Peng~3,Z.C.Zuo~4,H. M.Cui~3,Y.Wang~4,Z.H.Ren~4 (1 College of Veterinary Medicine,Sichuan Agricultural University,Yaan,Sichuan 625014,P.R. China,2 Department of Veterinary Medicine,Northeast Agricultural University,Harbin 150030, P.R.China,3Sichuan Key Laboratory of Animal Disease and Human Health,Yaan,Sichuan, 625014,P.R.China Sichuan 625014,P.R.China,4 Sichuan Key Laboratory of environmental hazards disease,Yaan,Sichuan 625014,P.R.China)

Zearalenone(ZEN) is an estrogenic mycotoxin produced by several Fusarium species,which can contaminate food and *** compounds elicit a wide spectrum of toxic effects,including the capacity to alter normal immune *** this study,the in vitro effects of the treatment of ConA-stimulated splenic lymphocytes with ZEN(0-25μg/mL) were *** modulates the expression of IL-2,IL-6,and IFN-γ. The IL-2 levels were up to four fold higher(P<0.05) compared with the levels in the control levels at toxin concentrations of 25μg/mL after 48 h of *** IL-6 levels were critically suppressed at this concentration;these changes were very statistically significant(P<0.05).At lower ZEN concentrations(0.1.0.4 and 1.6μg/mL), the IFN-γlevels changed slightly;however at 6.25 and 25μg/mL,the IFN-γresults reached statistical significance compared with the control levels(P<0.05).These data suggest that ZEN has potent effects on the expression of chicken splenic lymphocytes cytokines at the mRNA level.

关键词： zearalenone mycotoxin immunity chicken cytokines

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Nonclassic Major Histocompatibility Complex ClassⅠTake Capital Contribution to Reproduction of Dairy Cow

Nonclassic Major Histocompatibility Complex ClassⅠTake Capi...

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中国畜牧兽医学会家畜内科学分会第七届代表大会暨学术研讨会

作者： Lei Shu~1,Shen Zhang~1,Ganzhen Deng~(1*),Yue Wu~1,Mingyue He~1,Beibei Li~1,Chengye Li~1, Kechun Zhnag~3 (1 College of Veterinary Medicine,Huazhong Agricultural University.Wuhan,430070,China 2 Guangming Milk Product Cooperation Ltd,Shanghai,200436,China)

Major histocompability complex(MHC) takes capital contribution to maternal immune response to semiallogenic ***,the evidence of MHC I transcription on maternal peripheral blood monouclear cells(PBMC) and fetal tissues are still incompletely *** pairs of MHC-1 genes were chosen according to their homology and possible *** transcription levels on maternal PBMC from all three trimesters,and transcription levels in fetal tissues were compared to evaluate their contribution to cattle *** results suggested that MHC-1 gene composition and overall transcription levels on PBMC were not similar to that in fetal placenta in last *** geneⅤ(BoLA-NC3* 50201) upregulated throughout pregnancy,maybe act as an immune protective mechanism for pregnant cow;gene IV(BOLA-NC1*) transcription initialized fetus nidation;gene ID(HD15) transcription keeps the pregnancy up:GeneⅡ(MICB) transcription increases the immune response between fetus and dam,and then let fetus *** function of geneⅣsilenced in neonatal ear tissues was pended to further *** geneⅦ,ⅧandⅨshould be synergy with nonclassic gene V(BoLA-NC3*50201) to act as an immune protective mechanism for pregnant cow *** gene VI contribute to pregnancy establishment ant maintenance at first,and then to fetus expulsion *** the classic genes upregulated in neonatal ear tissues,suggest the prevention for fetus *** of heavy chain gene significantly downregulated in all dam and fetal tissues,the effect of classic MHC I genes is restricted.

关键词： diary cow MHC-I reproduction

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不同来源大肠杆菌中质粒介导的喹诺酮耐药基因(PMQR)调查研究

不同来源大肠杆菌中质粒介导的喹诺酮耐药基因(PMQR)调查研究

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中国畜牧兽医学会家畜传染病学分会第七届全国会员代表大会暨第十三次学术研讨会

作者：张云飞王红宁张安云高荣王霆邹立扣曾博四川大学生命科学学院四川大学“动物疫病防控与食品安全四川省重点实验室” 四川农业大学动物医学院雅安 625014

本试验旨在研究异源大肠杆菌对氟喹诺酮类药物(fluoroquinolones,FQs)的耐药表型和质柱介导FQs耐药基因(PMQR)的特征。2006-2008年从四川、河南等地的规模化猪场、医院及野生动物园分离的141异源大肠杆菌,采用K-B纸片法检测其对奈啶酸(... 详细信息

本试验旨在研究异源大肠杆菌对氟喹诺酮类药物(fluoroquinolones,FQs)的耐药表型和质柱介导FQs耐药基因(PMQR)的特征。2006-2008年从四川、河南等地的规模化猪场、医院及野生动物园分离的141异源大肠杆菌,采用K-B纸片法检测其对奈啶酸(NA)等六种FQs的耐药表型,并运用PCR方法检测PMQR基因:qnrA、qnrB、qnrS、aac(6′)-Ib-cr、qepA和qnrD.结果表明:试验菌株对各药物的耐药率分别为:萘啶酸(58.93%)、诺氟沙星(56.25%)、氧氟沙星(54.46%)、恩诺沙星(53.57%)、左旋氧氟沙星(51.79%)和环丙沙星(51.77%).其中,猪源及野生动物源菌株以对萘啶酸耐药为主,耐药率分别为72.22%和12.50%;人源菌株以对诺氟沙星耐药为主,耐药率为74.29%。PCR检测结果表明,PMQR基因的检出率由高到低依次为aac(6′)-Ib-cr(49.65%)、qnrD(23.40%)、qnrS(22.70%)、qnrB(21.28%)、qnrA(14.89%)及qepA(8.51%).猪源菌株中aac(6′)-Ib-cr和qnrD检出率高,分别为69.57%和43.47%.aac(6′)-Ib-cr和qnrS在人源菌株中分布较普遍,检出率分别为54.29%和37.14%.qnrA,qnrB和qepA在野生动物源菌株中均有一定检出率,分别为4.17%、8.33%和4.17%。本研究首次对不同来源大肠杆菌中六种PMQR基因进行调查研究,为大肠杆菌耐药性的防控提供参考依据.

关键词：大肠杆菌 FQs PMQR 耐药基因耐药表型

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13种消毒剂对规模化蛋鸡场环境微生物MIC、MBC测定

13种消毒剂对规模化蛋鸡场环境微生物MIC、MBC测定

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中国畜牧兽医学会家畜传染病学分会第七届全国会员代表大会暨第十三次学术研讨会

作者：康润敏王红宁高荣周英顺庞平四川大学“动物疫病与食品安全四川省重点实验室”四川大学生命科学学院四川农业大学四川大学生命科学学院

为评价13种消毒剂(白疫杀、百毒杀、过氧化氢、PVP碘、万仑菌清、新洁尔灭.复合酚、新大卫1、碘伏、84、百毒杀S、百毒强杀、菌毒灭)对规模化鸡舍环境微生物(大肠杆菌8099,金黄色葡萄球菌ATCC 6538,枯草杆菌黑色变种芽孢ATCC 9372以及... 详细信息

为评价13种消毒剂(白疫杀、百毒杀、过氧化氢、PVP碘、万仑菌清、新洁尔灭.复合酚、新大卫1、碘伏、84、百毒杀S、百毒强杀、菌毒灭)对规模化鸡舍环境微生物(大肠杆菌8099,金黄色葡萄球菌ATCC 6538,枯草杆菌黑色变种芽孢ATCC 9372以及环境微生物中的11株大肠杆菌,7株金黄色葡萄球菌与3株枯草芽孢杆)的消毒作用,采用肉汤稀释法进行了药物最低抑菌浓度(MIC)和最低杀菌浓度测定(MBC).结果表明:13种消毒剂药物大肠杆菌的MIC最低的依次为是过氧化氢23.4mR/L,PVP碘39-78.1 mg/L以及碘伏39.0-156.3 mg/L;对大肠杆菌MBC最低的依次为过氧化氢46.8 mg/L,PVP碘39-156.3mg/L和碘伏78.1-156.3 mg/L;对金黄色葡萄球菌的MIC最低依次为PVP碘39-78.1 mg/L,过氧化氢46.8 mg/L以及新洁尔灭和新大卫62.5mg/L;对金黄色葡萄球菌的MBC最低的依次为PVP碘39-78.1mg/L,过氧化氢46.8 mg/L,新大卫62.5 mg/L;对芽孢杆菌的MIC最低依次为新洁尔灭62.5mg/L,新大卫62.5mg/L,PVP碘78.1-156.3 mg/L;对芽孢杆菌的MBC最低依次为新洁尔灭62.5 mg/L,过氧化氢93.6 mg/L,新大卫125mg/L。结论:对于规模化鸡场对大肠杆菌抑菌效果最好的的过氧化氢,对金黄色葡萄球菌抑菌效果最好的是PVP碘,对于枯草芽孢杆菌抑菌效果最好的是新洁尔灭。

关键词：消毒剂最低抑菌浓度最小杀菌浓度

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Effects of deoxynivalenol on the mRNA levels of IL-2,IL-6,and IFN-γin the splenic lymphocytes of chickens

Effects of deoxynivalenol on the mRNA levels of IL-2,IL-6,an...

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中国畜牧兽医学会家畜内科学分会第七届代表大会暨学术研讨会

作者： Y.C.Wang~(1,2,3),J.L.Deng~(1,2,3*),S.W.Xu~(4,*),X.Peng~(1,2,3),Z.C.Zuo~(1,2,3),H.M.Cui~(1,2,3), Y.Wang~(1,2,3),Z.H.Ren~(1,2,3) (1 College of Veterinary Medicine,Sichuan Agricultural University,Yaan,Sichuan 625014,P.R. China,2 Sichuan Key Laboratory of Animal Disease and Human Health,Yaan,Sichuan,625014, P.R.China,3 Sichuan Key Laboratory of environmental hazards disease,Yaan,Sichuan 625014, P.R.China,4 Department of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,P.R.China)

The immunotoxicity of deoxynivalenol(DON) as well as the induction of cytokines and related genes in the splenic lymphocytes of chicken were *** the present study,the in vitro effects of DON(0-50μg/mL) treatment on ConA-stimulated splenic lymphocytes were examined after 48 *** modulates the expression of IL-2,IL-6,and IFN-γ.The IL-2 mRNA levels at 0.2.3.2,12.5,and 50μg/mL DON were significantly higher(P < 0.05) than the control group(ConA-only stimulated cells group).The highest IL-2 mRNA level was reached at 12.5μg/mL *** IL-6 levels were critically suppressed at all concentrations,and the differences from the control group(P < 0.05) were statistically ***-6 expression was highest at 12.5μg/mL *** IFN-γlevels were significantly higher(P < 0.05) than the control group,and IFN-γexpression was lowest at 0.8μg/mL *** data indicate the potent effects of DON on cytokine mRNA expression in chicken splenic lymphocytes.

关键词： Deoxynivalenol Mycotoxin Immunity Chicken Cytokines

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