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1.
为了解羊只布鲁氏菌疫苗(S2株)口服免疫的安全性及免疫效果,在甘肃省平凉市2个乡镇,各选1个行政村,选择散养户饲养的264只羊作为试验羊,进行小范围的布鲁氏菌S2株活疫苗口服免疫试验,其中3月龄以上羊只(包括怀孕母羊)每只口服活菌100×2亿,3月龄以下羊每只口服活菌100亿。分别在免疫后的30、60、180、210 d,采集试验羊血清样品,采用虎红平板凝集(RBT)+试管凝集联合试验(SAT)、酶联免疫吸附试验(ELISA)进行血清抗体检测。结果显示:试验组羊只免疫后均未出现异常反应,口服免疫30 d后,试验羊只2种检测方法检测的免疫抗体阳性率分别为47.3%和53.8%,不同时间2种方法的检测结果基本一致(P 0.05)。结果表明,布鲁氏菌S2株活疫苗按照试验剂量免疫安全有效,适用于所有月龄羊只以及怀孕母羊的整群免疫,可先行试点,逐步推广应用。 相似文献
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《中国预防兽医学报》2020,(3)
为评价不同商品化布鲁氏菌疫苗对小尾寒羊免疫后抗体消长规律及疫苗免疫效果,本研究选取3月龄～6月龄的羔羊随机分为5组,分别为布鲁氏菌疫苗S2口服组(70只)、A19皮下注射组(30只)、M5皮下注射组(70只)、M5口服组(70只)和对照组(30只),并采用ELISA方法检测疫苗免疫后不同时间段羊血清中IgM和IgG抗体的消长规律,用虎红平板凝集试验(RBPT)和ELISA方法检测疫苗免疫后不同时间段羊血清中抗体转阳率和抗体持续期的变化趋势,统计分析其抗体消长规律。结果显示,免疫前,所有实验羊均未检测到抗体,不同疫苗免疫后羊产生的IgM抗体水平最高的是A19皮下注射组,最低的是M5口服组,在免疫7 d～14 d内4组实验组羊的IgM抗体水平均达到峰值,其抗体持续时间在45 d～60 d。不同疫苗免疫后羊产生IgG抗体水平最高的是M5皮下注射组,最低的是M5口服组,在免疫7 d～28 d内4组实验组羊的IgG抗体水平均达到峰值,其抗体持续时间在60 d以上。M5皮下注射组、A19皮下注射组、M5口服组和S2口服组羊在免疫7 d～14 d内抗体转阳率均达到最大值:采用RBPT方法检测抗体转阳率分别为67%、56%、41%和45%,采用ELISA方法检测其抗体转阳率分别为71%、56%、37%和38%,其抗体水平持续时间约为150 d。抗体水平结果表明,A19疫苗可以用于羊布鲁氏菌病的免疫,M5疫苗皮下注射免疫羊的效果最好,口服S2的疫苗免疫效果要优于口服M5疫苗。本研究为制定羊布鲁氏菌病合理的疫苗免疫方案提供了科学依据。 相似文献
3.
为建立一种羊种布鲁氏菌Rev.1疫苗株PCR-RFLP鉴定方法,利用羊种布鲁氏菌Rev.1具有链霉素抗性的特性,选取与链霉素抗性相关编码核糖体蛋白S12的rps L基因为模板设计引物,经PCR扩增后,将产物进行Nci I酶切,发现羊种布鲁氏菌疫苗株Rev.1出现约500 bp条带,而不具有链霉素抗性的羊种布鲁氏菌株16M则出现384 bp条带。结果表明,PCR-RFLP方法能够用于羊种布鲁氏菌Rev.1疫苗株的鉴定,这为今后区分羊种布鲁氏菌疫苗株Rev.1免疫与野株感染提供了基础。 相似文献
4.
布鲁氏菌外膜蛋白OMP15.6表达及免疫反应原性测定 总被引：2，自引：0，他引：2
表达羊布鲁氏菌16M预测外膜蛋白OMP15.6,探索其作为诊断抗原的可能性。采用降落PCR方法,从羊布鲁氏菌16M基因组DNA中扩增出423bp的基因片段,将该片段克隆于原核表达载体PGEX-4T-2,构建重组表达载体。经IPTG诱导,SDS-PAGE检测,结果表明,在大肠杆菌中成功表达了外膜蛋白OMP15.6。经West-ern-blotting检测,该蛋白能与豚鼠抗布鲁氏菌阳性血清发生特异性免疫反应,为其之后的抗原性以及免疫原性研究奠定了良好的基础。 相似文献
5.
本文通过对S2株布鲁氏菌活疫苗免疫效果实验及防控建议进行了阐述,发现采用正常剂量和加强剂量免疫羊血清抗体的阳转率明显低于加倍剂量免疫羊的血清抗体的阳转率。为此在免疫的过程中要严格按照S2株布鲁氏菌活疫苗说明书进行操作,确保免疫结果达到国家要求标准,在布病防控中,各部门加强合作,坚持以人为本,加强个人防护,对患病人员积极治疗,最大限度的减轻农牧民的经济负担。 相似文献
6.
为研究羊免疫布鲁氏菌病基因缺失活疫苗(M5-90Δ26株)和弱毒疫苗(S2株)效果,选取220只3～12月龄的布病阴性羊,随机分为3组,免疫48h内观察疫苗副反应,并在免疫7d、14d、30d、90d后采血进行抗体检测。结果显示：免疫48h内,M5-90Δ26注射组和S2口服组均未出现发烧症状,M5-90Δ26注射组1只试验羊出现采食量下降,S2口服组未见羊群采食量下降。M5-90Δ26注射组免疫7d抗体阳性率97%,免疫14d和30d抗体阳性率100%;S2口服组免疫后7d抗体阳性率49%,免疫30d抗体阳性率最高(81%);对照组未检测到布病抗体阳性。综上所述,布病M5-90Δ26在免疫后抗体水平上优于S2株,但可能存在副反应,建议免疫前观察羊状态,严格按照疫苗说明书进行免疫。 相似文献
7.
[目的]探究湖羊经布鲁氏菌S2疫苗免疫后的血清抗体消长规律。[方法 ]采用皮下注射25亿CFU(正常剂量)、口服免疫100亿CFU(正常剂量)、口服免疫150亿CFU(1.5倍剂量)3种免疫方法,对试验湖羊进行S2疫苗接种。在免疫前后的不同时间点,采集血液,分离血清,通过虎红平板凝集试验(RBT)和试管凝集试验(SAT)检测血清抗体水平。[结果 ]皮下免疫组在免疫后7 d出现抗体,21 d达到峰值,平均抗体效价最高为1:400,RBT和SAT抗体阳性率均为100%;免疫后120 d,RBT、SAT血清抗体阳性率分别为53%、56%;血清抗体存续时间为240 d。1.5倍剂量口服组免疫后21 d,RBT、SAT抗体阳性率分别为80%、83%,平均抗体效价最高为1:230.9,血清抗体存续时间为150 d。正常剂量口服免疫组在免疫后14 d产生抗体,21 d的抗体阳性率最高,RBT、SAT抗体阳性率分别为50%、57%,平均抗体效价最高为1:156,血清抗体存续时间为120 d。[结论 ]布鲁氏菌S2疫苗皮下免疫效果明显优于口服免疫效果,但可能存在免疫副反应;1.5倍剂量口服优于正常剂量口服。 相似文献
8.
为探究多浪羊布鲁氏菌活疫苗(M5)的免疫抗体消长规律,在新疆南疆地区某行政村,选择100只多浪羊进行皮下注射布鲁氏菌活疫苗(M5)免疫试验。免疫前,对所有试验羊只全部采血,分别于免疫后30、90、180 d,随机采集30只羊全血,分离血清,用虎红平板凝集试验(RBT)、试管凝集试验(SAT)进行布鲁氏菌抗体检测。结果显示:免疫前,所有试验羊只均未检测到抗体,免疫30 d后,73.33%的羊检测到抗体,90d后只有13.30%的羊还能检测到抗体,180 d后未检测到抗体。结果表明,布鲁氏菌活疫苗(M5)能够使多浪羊产生良好的免疫反应,免疫抗体持续期约为180 d。这说明对免疫180 d后的多浪羊进行布鲁氏菌抗体检测,可为区分自然感染和免疫提供参考。 相似文献
9.
利用细胞总RNA提取试剂盒,从1月龄SPF鸡白细胞中分离提取总RNA,经过反转录PCR(RT-PCR)扩增出鸡β-防御素(Gal-1)cDNA片段。PCR产物经凝胶回收纯化后与pMD18-T载体连接,转化感受态JM109细胞,在含X-gal、IPTG的LB平板上筛选阳性克隆,经菌落PCR与质粒限制性酶切鉴定后,对目的片段进行测序。结果表明RT-PCR扩增出的cDNA片段ORF大小为198bp,用Blast程序进行检索比较表明该序列与Genbank中发表的鸡Gal-1 cDNA编码区序列同源性高达99.5%。该cDNA序列编码65个氨基酸,包括信号肽、前片段和成熟肽,成熟肽由40个氨基酸残基组成。 相似文献
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11.
Bhide M Chakurkar E Tkacikova L Barbuddhe S Novak M Mikula I 《Veterinary microbiology》2006,112(1):33-41
Johne's disease is one of the main causes of economic losses in ruminants and a major health hazard in the developing and developed world. Up till now, many microbiological, serological and molecular methods have been tried for the detection of Mycobacterium avium subsp. paratuberculosis (MAP). In this study, we attempt a PCR-based detection of IS900, distinct insertion sequences of MAP from the buffy coat of cattle (n=262) and sheep (n=78), and direct genotyping by single strand conformational polymorphism (SSCP). A total of 30 (11.45%) cattle and one sheep (1.28%) were positive for MAP-IS900. This IS900-based PCR detection proved highly specific, particularly when tested on other non-MAP strains. SSCP analysis grouped the MAP-IS900 into four distinct clusters based on different band patterns. Nucleotide sequence variability between MAPs detected from sheep (GenBank accession ) and cattle (GenBank accession -) was noticed in the study. Although, in recent years IS900-PCR-based detection of MAP from WBCs is being used in human, its use in animals is still limited. Our work not only supports its use in animals but also suggests further IS900-SSCP-based MAP-genotyping, coupled with DNA sequencing, as a promising tool for rapid and effective Johne's disease surveillance. 相似文献
12.
Pam Dachung Luka Chrisostom Ayebazibwe David Shamaki Frank Norbert Mwiine Joseph Erume 《Journal of veterinary science (Suw?n-si, Korea)》2012,13(3):323-325
Peste des petits ruminants (PPR) diagnosis from suspected samples from sheep and goats was carried out. Buffy coat, tissues, and oculo-nasal swabs were analyzed using nucleoprotein (NP3/NP4) and fusion protein (F1/F2) gene primers, respectively. Analysis of the sample types and primer set revealed that buffy coat are the best type of samples for PPR diagnosis and the use of two set of primers will increase the number of positives. 相似文献
13.
Whole blood, buffy coat and plasma from sheep infected with tick-borne fever (TBF) were inoculated into embryonated hen eggs, primary chicken embryo fibroblasts, primary foetal lamb kidney cells, mouse L cells, and ovine pulmonary macrophage cultures. Infected buffy coat cells were fused with primary chicken embryo fibroblasts, mouse L cells, ovine pulmonary macrophages and BHK-21 cells, using UV irradiated Sendai virus. Evidence of multiplication was not obtained. 相似文献
14.
为了对鸡输卵管特异表达载体表达的重组人溶菌酶(rhLYZ)的理化及生物学特性进行鉴定,用直接结晶法从重组载体注射鸡蛋清中提取溶菌酶,以兔抗hLYZ血清为一抗,HRP-标记的羊抗免IgG为二抗,进行Western blotting检测,结果表明表达在鸡蛋清中的rhLYZ能被hLYZ特异抗体识别;将纯化的rhLYZ和天然hLYZ在不同温度的水浴中孵育30min,用RBB-R染料标记比色法检测溶菌酶活性的变化,结果显示二者的热稳定性无显著差异;将纯化的rhLYZ和天然hLYZ用不同PH的溶液稀释,40C孵育30min后进行酶的活性测定,结果显示两者在pH3.0～11.0范围内具有活性,最强活性pH为7.0;以12种常见细菌为试验对象,用最小抑菌浓度法测定含rhLYZ蛋清和纯化rhLYZ的溶菌活性,结果显示两者对鲫鱼嗜水气单胞菌、变形杆菌、枯草杆菌、无鞭毛伤寒杆菌、白色念珠菌和柠檬色葡萄球菌具有很强的抑制作用,对痢疾杆菌、白色葡萄球菌、金黄色葡萄球菌和大肠杆菌具有较强的抑制作用,对有鞭毛伤寒杆菌和鳖嗜水气单胞菌无明显的抑制作用.这些试验结果显示,表达在重组载体注射鸡蛋清中的rhLYZ具有与天然hLYZ十分相似的相对分子质量、热稳定性、pH活性范围和溶菌谱. 相似文献
15.
The aims of this investigation were to determine the prevalence of ovine herpesvirus type 2 (OvHV-2) (the causative agent of malignant catarrhal fever) infection in cattle, the carrier status of sheep and goats, and to define the pattern of acquisition of OvHV-2 in lambs under natural flock conditions in Kashmir, India. None of the buffy coat samples from 21 lambs contained OvHV-2 DNA sequences up to 28 days after birth, only one lamb had sequences of OvHV-2 DNA as early as 29 days after birth, and they were detected in the other 20 lambs when they were between 43 and 94 days of age. Sequences of OvHV-2 DNA were detected in buffy coat samples from 28 (85 per cent) of 33 adult sheep and in 16 (61 per cent) of 26 samples from adult goats by hemi-nested PCR. Seventeen (31 per cent) of 55 cattle with malignant catarrhal fever-like clinical signs had sequences of OvHV-2 DNA in their blood, and nine of the 17 died, all of them during the months of April to November, between November 2002 and March 2004. No clinical cases of sheep-associated malignant catarrhal fever was recorded during the months of December to March. The overall prevalence of OvHV-2 infection in the cattle in the region was estimated to be less than 1 per cent. 相似文献
16.
Unraveling avian and reptilian hematology: An optical and electron microscopic study of the buffy coat 
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下载免费PDF全文 Filipe Fontes Pinto Célia Lopes Fernanda Malhão Ricardo Marcos 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2018,47(3):407-414
Background
Blood centrifugation and buffy coats are at the cornerstone of hematology. In mammals, the buffy coat has a layered disposition (from bottom to top) with neutrophils on top of erythrocytes, followed by monocytes/lymphocytes, and platelets. In nonmammals, this distribution is unknown. Recently, the cell tube block (CTB) technique was developed to study the buffy coat, but it was never applied to nonmammal buffy coats.Objectives
This study aimed to evaluate using the CTB technique to study reptilian and avian buffy coats and to propose its use for clinical applications.Methods
Blood from five birds and eight reptiles of different species was obtained to make CTBs that were processed for optical/electron microscopy. H&E, Sirius red, and immunohistochemistry staining against CD3 (to label T lymphocytes) were applied to the CTBs.Results
In birds, the buffy coat had a layered appearance with the granulocyte layer containing granulocytes (heterophils and eosinophils) and nucleated erythrocytes followed by a mononuclear cell layer containing lymphocytes, monocytes, and thrombocytes. In some animals, a nucleated erythrocyte layer was observed admixed with the granulocyte/mononuclear cell layer. A small clot within the buffy coat was seen in seven reptiles, and less definition of layers occurred in reptiles, with only one or two layers. Lymphocytes appeared toward the top of the buffy coat.Conclusions
From a comparative hematology perspective, the buffy coat of mammals differs from that of birds and more from that of reptiles. The CTB technique can be used to study these differences in avian and reptilian hematology, especially to study atypical circulating cells, hemoparasites, or blood cell proportions in health and disease. 相似文献17.
De Las Heras M Ortín A Salvatori D Pérez de Villareal M Cousens C Miguel Ferrer L Miguel Cebrián L García de Jalón JA Gonzalez L Michael Sharp J 《Research in veterinary science》2005,79(3):201-264
Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring contagious lung neoplasia caused by jaagsiekte sheep retrovirus (JSRV). Although no specific circulating antibodies against the virus can be detected in infected sheep, JSRV proviral DNA sequences can be found in peripheral blood leukocytes (PBLs) in clinically affected and in a proportion of in contact animals. In this study, existing hemi-nested PCR procedure is compared with a new one-step PCR technique that was developed to minimise potential DNA contamination and reduce sample and reagent handling. Different blood preparations were assessed and the best results were achieved on DNA prepared from buffy coat. The sensitivity of this PCR was lower in JSRV infected sheep without lesions of OPA than in clinically affected sheep, which indicate that this PCR may not be not fully appropriate for screening of individual sheep, but rather to provide results at flock level. This PCR is the only currently available blood test for detection of JSRV infected sheep and may be useful in epidemiological studies and in control programmes of OPA. 相似文献
18.
OBJECTIVE: To evaluate the buffy coat and apheresis methods for preparation of platelet concentrates from equine blood by comparing platelet and growth factor concentrations. ANIMALS: 15 mature mixed-breed geldings. PROCEDURE: Whole blood samples were collected and processed by use of a buffy coat or apheresis method to obtain platelet poor and platelet concentrated fractions. The PCV, WBC count, and platelet count were compared among whole blood samples, platelet poor fractions, concentrates obtained by use of the apheresis method (ie, apheresis platelet concentrates), and concentrates obtained by use of the buffy coat method (ie, buffy coat platelet concentrates). Concentrations of transforming growth factor-beta (ie,TGF-beta1 andTGF-beta2) and insulin-like growth factor were compared between buffy coat and apheresis platelet concentrates. RESULTS: Platelet concentrations were 8.9-fold and 5.2-fold greater in buffy coat and apheresis platelet concentrates, respectively, compared with whole blood. Platelet concentrations were 13.1-fold greater in filtered apheresis platelet concentrates, compared with whole blood. TGF-beta1 concentrations were 2.8-fold and 3.1-fold greater in buffy coat and apheresis platelet concentrates, respectively, and TGF-beta1 concentrations were 10.5-fold greater in filtered apheresis platelet concentrates, compared with whole blood. TGF-beta2 concentrations were 3.6-fold greater in apheresis platelet concentrates, compared with whole blood. Platelet concentrations correlated with growth factor concentrations across all blood and platelet fractions. White blood cell counts had a significant positive correlation with TGF-beta1 concentration in buffy coat platelet concentrates. CONCLUSIONS AND CLINICAL RELEVANCE: Platelets and TGF-beta1 can be concentrated reliably from equine blood by use of buffy coat or apheresis methods, without modification of the protocols used for humans. 相似文献
19.
用直接结晶法从重组载体注射鸡蛋清中提取重组人溶菌酶（rhLYZ），并对其理化及生物学特性进行了鉴定。结果显示，rhLYZ能被hLYZ特异抗体识别；纯化的rhLYZ和天然hLYZ的热稳定性无显著差异，二者在pH3．0～11．0范围内均具有活性，pH为7．0时活性最强。采用最小抑菌浓度法测定了含rhLYZ蛋清和纯化rhLYZ对12种常见细菌的溶菌活性，二者对除有鞭毛伤寒杆菌和鳖嗜水气单胞菌以外的参试茵均有不同程度的抑制作用。表明，表达在重组栽体注射鸡蛋清中的rhLYZ具有与天然hLYZ十分相似的分子质量、热稳定性、pH活性范围和溶菌谱。 相似文献
20.
Frequency of persistent bovine viral diarrhea virus infection in selected cattle herds 总被引：5，自引：0，他引：5
Sera and blood buffy coat samples were obtained from 3,157 cattle in 66 selected herds. Antibodies to bovine viral diarrhea (BVD) virus were detected in 89% of the serum samples by immunoprecipitation or virus-neutralization tests. Cytopathic or noncytopathic BVD viruses were isolated from blood buffy coat samples from 60 cattle in 6 herds. A second blood buffy coat sample was obtained from 54 of the 60 cattle 2 months after the initial sampling, and BVD virus was isolated again from each cow. The 54 cattle were considered persistently infected with BVD virus. The frequency of persistent infection was 1.7%. 相似文献