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Pattern of Eimeria oocyst output and repeatability in naturally infected suckling Rhön lambs. 自然感染Rhön哺乳羔羊的艾美耳球虫卵囊输出模式和可重复性。
Pub Date : 2001-11-05 DOI: 10.1046/J.1439-0450.2001.00493.X
M. Gauly, C. Krauthahn, C. Bauer, G. Erhardt
The faeces of 14 Rhön lambs were examined every second day between 7 and 99 days of age for the presence of Eimeria oocysts. Eimeria absata, E. bakuensis, E. faurei, E. granulosa, E. intricata, E. ovinoidalis, E. pallida, E. parva and E. crandallis/weybridgensis were identified. The predominant species were E. ovinoidalis, E. parva, E. crandallis/weybridgensis and E. bakuensis. Using a statistical model, the oocyst excretion rate was described as a sequence of periods with decreasing levels and varying length ('excretion periods') interrupted by intervals with no or very low oocyst counts. Several variables could be deduced from these two parameters, including the length of an excretion period and the maximum output during an excretion period. Thc estimated repeatability for oocyst counts for the different species ranged from 0.05 to 0.41. This result provides a starting point for possible genetic selection based on faecal oocyst counts of Rhön sheep for resistance to Eimeria infections.
在7 ~ 99日龄期间,每隔一天对14只Rhön羔羊的粪便进行艾美耳球虫卵囊检查。鉴定出阿布艾美耳螨、巴库艾美耳螨、山羊艾美耳螨、颗粒艾美耳螨、复杂艾美耳螨、山羊艾美耳螨、苍白斑艾美耳螨、细小艾美耳螨和大布氏艾美耳螨。优势种为野伊蚊、小伊蚊、大伊蚊/韦氏伊蚊和巴库伊蚊。使用统计模型,将卵囊排泄率描述为一系列水平下降且长度变化的周期(“排泄期”),其间间隔无卵囊计数或卵囊计数极低。从这两个参数可以推断出几个变量,包括排泄期的长度和排泄期的最大产量。不同种类的卵囊计数的估计重复性在0.05 ~ 0.41之间。这一结果为基于Rhön羊粪便卵囊计数的艾美耳球虫感染抗性的可能遗传选择提供了一个起点。
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引用次数: 20
Field study on the efficacy of two different vaccination schedules with HYORESP in a Mycoplasma hyopneumoniae-infected commercial pig unit. 在猪支原体肺炎感染的商品猪单位中,两种不同的HYORESP疫苗接种计划的效果的实地研究。
Pub Date : 2001-11-05 DOI: 10.1046/J.1439-0450.2001.00494.X
S. Kyriakis, C. Alexopoulos, J. Vlemmas, K. Sarris, S. Lekkas, M. Koutsoviti-Papadopoulou, K. Saoulidis
A trial was carried out with HYORESP a Mycoplasma hyopneumoniae (M. hyo) vaccine in order to confirm the benefit of vaccination under field conditions in a commercial industrial farrow-to-finish unit, contaminated with M. hyo. Infection with M. hyo was confirmed through positive blood and colostrum samples [enzyme-linked immunosorbent assay (ELISA) test] combined with positive gross lesions of the lung at slaughter. Two different vaccination schedules were tested. Pigs were randomly allocated to three groups: control non-vaccinated group (n = 130, given a placebo injection at 3, 25 and 70 days of age); early vaccinated group (n = 128, given vaccination at 3 and 25 days of age and a placebo at 70 days of age); late vaccinated group (n = 132, given a placebo at 3 and 25 days of age and vaccination at 70 days of age). Both growth rate and feed conversion ratio were signifcantly (P < 0.05) improved in the vaccinated groups compared with the control group. The lung lesion score was also significantly (P < 0.05) improved in both vaccinated groups. In this trial, it was clearly demonstrated that vaccination is highly effective in improving performance in pig units infected with M. hyo. The improvement in the feed conversion ratio in the vaccinated groups was especially impressive: -0.411 (13% improvement) in the group vaccinated twice at 3 and 25 days of age; -0.162 (5% improvement) in the group vaccinated once at 70 days of age. Performances were better when two shots were given early in life compared with one shot later--probably due to an infection taking place rather early in life for most of the pigs. Moreover, a significant reduction in the cost of supportive (injectable) medication was noticed in vaccinated pigs. In conclusion, HYORESP proved to be a very efficacious tool to control M. hyo in infected herds with its remarkable flexibility that allows the vaccination schedule to be adapted to the specific field conditions.
对HYORESP一种猪肺炎支原体(支原体)疫苗进行了试验,以确认在被支原体污染的从产仔到肥育的商业工业单位的现场条件下接种疫苗的益处。通过血液和初乳样本(酶联免疫吸附试验(ELISA))阳性,并在屠宰时肺部大体病变阳性,证实感染了猪支原体。测试了两种不同的疫苗接种时间表。将猪随机分为3组:未接种疫苗对照组(n = 130头,分别在3、25和70日龄注射安慰剂);早期接种组(n = 128,在3日龄和25日龄接种疫苗,在70日龄接种安慰剂);晚接种组(n = 132,在3日龄和25日龄给予安慰剂,在70日龄接种疫苗)。与对照组相比,接种组的生长率和饲料系数均显著(P < 0.05)提高。两组患者肺病变评分均显著提高(P < 0.05)。在这项试验中,清楚地表明,疫苗接种是非常有效的提高猪单位的性能感染猪分枝杆菌。接种疫苗组饲料系数的提高尤为显著:3日龄和25日龄接种两次的组饲料系数提高了-0.411 (13%);-0.162(改善5%),在70日龄接种一次疫苗的组。早期注射两次的猪比晚期注射一次的猪表现更好,这可能是由于大多数猪的感染发生在生命的早期。此外,在接种疫苗的猪中,支持性(注射)药物的成本显著降低。总之,HYORESP被证明是一种非常有效的控制受感染畜群中hym . hyo的工具,其显著的灵活性使疫苗接种计划能够适应特定的田间条件。
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引用次数: 24
Comparison of closely related orthopoxvirus isolates by random amplified polymorphic DNA and restriction fragment length polymorphism analysis. 近缘正痘病毒分离株随机扩增多态性DNA与限制性内切片段长度多态性分析比较。
Pub Date : 2001-11-05 DOI: 10.1046/J.1439-0450.2001.00484.X
M. Stemmler, H. Neubauer, H. Meyer
The reliability and reproducibility of random amplified polymorphic DNA analysis (RAPD) was compared with restriction fragment length polymorphism (RFLP) by analysing three virus strains isolated from zoo animals in Berlin and three isolates which were cultivated from pets from Northern Germany. The RAPD technique was evaluated as a reliable tool with good reproducibility of the patterns for each virus strain investigated. Problems of interpretation due to inconsistent intensity of bands in different polymerase chain reaction runs may arise for less experienced personnel. The RAPD analysis can be performed within one working day and needs less DNA compared with RFLP so costs will be reduced. The obvious advantage of RFLP is that the pattern can be traced to the recognition site of the restriction enzyme whereas the RAPD primer sequence is not present in the orthopoxvirus genome at all. To the authors knowledge, the RAPD technique has never been applied in DNA viruses before and they conclude that this technique is a useful tool for the discrimination of closely related cowpoxviruses.
通过对柏林动物园动物分离的3株病毒株和德国北部宠物培养的3株病毒株进行分析,比较了随机扩增多态性DNA分析(RAPD)与限制性片段长度多态性(RFLP)的可靠性和重复性。RAPD技术被评价为一种可靠的工具,对所调查的每种病毒株具有良好的重现性。在不同的聚合酶链反应运行中,由于条带强度不一致,可能会对经验不足的人员产生解释问题。RAPD分析可以在一个工作日内完成,与RFLP相比,需要更少的DNA,因此降低了成本。RFLP的明显优势是其模式可以追溯到限制性内切酶的识别位点,而RAPD引物序列根本不存在于正瘟病毒基因组中。据作者所知,RAPD技术以前从未在DNA病毒中应用过,他们得出结论,该技术是一种有用的工具,用于密切相关的牛痘病毒的区分。
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引用次数: 8
Occurrence of an invertebrate iridescent-like virus (Iridoviridae) in reptiles. 无脊椎动物中虹彩样病毒(虹彩病毒科)在爬行动物中的发生。
Pub Date : 2001-11-05 DOI: 10.1046/J.1439-0450.2001.00495.X
F. Just, S. Essbauer, W. Ahne, S. Blahak
Viral isolates were obtained in 1998, 1999 and 2000 from the lung, liver and intestine of two bearded dragons (Pogona vitticeps) and a chameleon (Chamaeleo quadricornis) and from the skin of a frill-necked lizard (Chamydosaurus kingii) by using viper heart cells (VH2) at 28 degrees C. Electron microscopic examination of infected VH2 cells revealed the assembly of icosahedral iridovirus-like particles measuring 139 nm (side to side) and 151 nm (apex to apex). Negatively stained virus particles had dimensions of 149 nm (side to side) and 170 nm (apex to apex). Polymerase chain reaction (PCR) amplification of purified viral DNA with primers corresponding to the partial gene encoding the major capsid protein (MCP) of Frog viris-3 (FV-3), the type species of the genus Ranavirus, was unsuccessful. In contrast, primers corresponding to the partial MCP gene of Chilo iridescent virus (CIV; genus Iridovirus) amplified 500-bp products with 97% identity to the nucleotide sequence of CIV and 100% identity to the nucleotide sequence of Gryllus bimaculatus iridescent virus (GbIV), an invertebrate iridescent virus. Virus protein profiles analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and restriction fragment length profiles of purified viral DNA treated with the endonucleases EcoRI, HindIII and HpaII were identical to those of GbIV.
1998年、1999年和2000年分别从两种胡须龙(Pogona vitticeps)和一种变色龙(Chamaeleo quadricornis)的肺、肝和肠以及在28℃下使用毒蛇心脏细胞(VH2)从一种褶颈蜥蜴(Chamydosaurus kingii)的皮肤中分离出病毒,对感染的VH2细胞进行电镜检查,发现组装了尺寸为139 nm(侧面)和151 nm(尖端)的二十面体虹膜病毒样颗粒。阴性染色的病毒颗粒尺寸分别为149 nm(侧对侧)和170 nm(端对端)。用蛙病毒3型(Frog virus -3, FV-3)主要衣壳蛋白(MCP)部分基因编码的引物对纯化的病毒DNA进行聚合酶链反应(PCR)扩增失败。相比之下,奇洛荧光病毒(CIV)部分MCP基因对应的引物;扩增的500 bp产物与CIV核苷酸序列的一致性为97%,与无脊椎动物虹膜病毒(Gryllus biaculatus iridescent virus, GbIV)核苷酸序列的一致性为100%。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析的病毒蛋白谱和经EcoRI、HindIII和HpaII内切酶处理的纯化病毒DNA的限制性内切片段长度谱与GbIV相同。
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引用次数: 37
Characterization of necrotoxigenic Escherichia coli (NTEC) strains isolated from healthy calves in Poland. 从波兰健康犊牛中分离的坏死性产毒素大肠杆菌(NTEC)菌株的鉴定
Pub Date : 2001-11-05 DOI: 10.1046/J.1439-0450.2001.00481.X
J. Osek
Faecal samples from 132 healthy, 4-8-week-old calves from four different farms were examined for necrotoxigenic Escherichia coli (NTEC) producing the cytotoxic necrotizing factors type 1 (CNF1) and type 2 (CNF2). CNF2 genes were detected by polymerase chain reaction in 24 (6.1%) of the 396 E. coli strains tested; these strains were found in 18 (13.6%) calves used in the study. None of the 396 E. coli isolates examined possessed the gene encoding CNF1. Overall, 28.8% of E. coli examined expressed the F17 fimbrial antigen. A strong association between CNF2 toxin and F17 fimbriae was found (62.5% of CNF2-positive strains were F17-positive). Moreover, six out of 24 NTEC strains had the Stx1 or the Stx2 shiga toxin genes, and three additional isolates possessed the eae genetic marker of the intimin protein.
对来自4个不同养殖场的132头4-8周龄健康犊牛的粪便样本进行了产坏死性大肠杆菌(NTEC)的检测,检测其产生细胞毒性坏死性因子1型(CNF1)和2型(CNF2)。396株大肠杆菌中24株(6.1%)经聚合酶链反应检测出CNF2基因;在研究中使用的18头小牛(13.6%)中发现了这些菌株。检测的396株大肠杆菌中没有一株具有编码CNF1的基因。总体而言,28.8%的大肠杆菌表达F17菌毛抗原。结果表明,CNF2毒素与F17菌毛有较强的相关性(62.5%的CNF2阳性菌株为F17阳性)。此外,24株NTEC菌株中有6株具有Stx1或Stx2志贺毒素基因,另外3株具有内膜蛋白eae遗传标记。
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引用次数: 14
Informative value of an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of bovine viral diarrhoea virus (BVDV) antibodies in milk. 间接酶联免疫吸附试验(ELISA)检测牛奶中牛病毒性腹泻病毒(BVDV)抗体的信息价值。
Pub Date : 2001-11-05 DOI: 10.1046/J.1439-0450.2001.00497.X
F. Beaudeau, C. Belloc, H. Seegers, S. Assié, P. Pourquier, A. Joly
Bulk and individual milk samples from 117 herds located in Brittany (west France) were used to assess: (i) the performance characteristics of an indirect enzyme-linked immunosorbent assay (ELISA) applied to individual milk for the detection of antibodies to bovine viral diarrhoea virus (BVDV); and (ii) the relationship between the bulk milk result obtained from this test and the within-herd prevalence of antibody-positive lactating cows. This ELISA test was based on a monoclonal antibody directed against non-structural protein NS2-3 of pestiviruses. At the individual level, based on 1113 matched milk/serum samples, the sensitivity and specificity of this test applied to milk, compared with the virus neutralization test on serum, were 95.0 and 97.7%, respectively. At the herd level, the relationship between the optical density percentage (OD%) of bulk milk and the within-herd prevalence of antibody-positive lactating cows was assessed using the receiver operating characteristics (ROC) analysis. Classes of OD% of bulk milk were determined so that they were associated with minimum intraclass and maximum between-class variances of within-herd prevalence of antibody-positive cows. The ROC analysis resulted in two classes of bulk milk results corresponding to different expected levels of within-herd prevalence. Herds with an OD% of bulk milk < 75% and > or = 75% had a mean observed prevalence of antibody-positive cows of 8.9 and 60.6%, respectively. Herds with a bulk milk result < 75% were expected to be BVDV free, whereas large variations in prevalence of antibody-positive cows existed in the herds with OD% > or = 75%. The test described in this study is suitable to identify herds likely to have a low prevalence of BVDV antibody-positive cows.
来自布列塔尼(法国西部)117个畜群的散装和单个牛奶样本用于评估:(i)用于检测牛病毒性腹泻病毒(BVDV)抗体的用于单个牛奶的间接酶联免疫吸附试验(ELISA)的性能特征;(ii)从该测试中获得的散装牛奶结果与牛群内抗体阳性的泌乳奶牛流行率之间的关系。该酶联免疫吸附试验基于一种针对鼠疫病毒非结构蛋白NS2-3的单克隆抗体。在个体水平上,基于1113份匹配的牛奶/血清样本,与血清病毒中和试验相比,该试验对牛奶的敏感性和特异性分别为95.0和97.7%。在牛群水平上,采用接受者工作特征(ROC)分析,评估散装奶光密度百分比(OD%)与奶牛群内抗体阳性发生率之间的关系。确定了散装牛奶OD%的等级,使其与抗体阳性奶牛群内流行率的最小类内方差和最大类间方差相关。ROC分析得出两类散装牛奶的结果,对应于不同的预期水平的群内流行率。散乳OD% < 75%和>或= 75%的牛群,抗体阳性奶牛的平均患病率分别为8.9%和60.6%。产奶量< 75%的奶牛被认为是无BVDV的,而产奶量>或= 75%的奶牛抗体阳性的流行率差异很大。本研究中描述的测试适用于确定BVDV抗体阳性奶牛可能低流行率的牛群。
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引用次数: 19
Efficacy of tilmicosin phosphate (Pulmotil premix) in feed for the treatment of a clinical outbreak of Actinobacillus pleuropneumoniae infection in growing-finishing pigs. 饲料中添加磷酸替米科星(Pulmotil预混料)治疗生长育肥猪临床爆发胸膜肺炎放线杆菌感染的效果。
Pub Date : 2001-11-05 DOI: 10.1046/J.1439-0450.2001.00492.X
G. Hoflack, D. Maes, B. Mateusen, M. Verdonck, A. de Kruif
A double-blind randomized clinical trial was carried out to investigate the efficacy of tilmicosin (Pulmotil premix) for the treatment of a clinical outbreak of Actinobacillus pleuropneumoniae infection in growing-finishing pigs. The effects of tilmicosin administration in the feed at 400 mg/kg and an injection therapy of clinically diseased pigs with long-acting oxytetracycline (Terramycine LA) at 20 mg/kg bodyweight were compared. Both groups, totalling 147 pigs, were compared during a medication period of 15 days and a post-medication period of 11 days by means of different clinical and performance parameters. During the medication period, the tilmicosin group showed a significant advantage with respect to the number of new disease cases (P < 0.01), and a non-significant advantage regarding the number of removed pigs (P = 0.16), the number of sick pigs that recovered (P = 0.27) and the time to recovery (P = 0.42). During the post-medication period, the pigs of the tilmicosin group showed numerical non-significant benefits (P > 0.05) with respect to the clinical parameters. During the overall study period (26 days), the average daily gain and the feed conversion ratio were both significantly (P < 0.01) better in pigs from the tilmicosin group compared with pigs from the oxytetracycline group. This study demonstrated that in-feed medication of tilmicosin at a dosage of 400 mg/kg is efficacious for the treatment of a clinical respiratory disease outbreak of A. pleuropneumoniae infection in growing-finishing pigs. Compared with oxytetracycline injection of clinically diseased pigs, the tilmicosin treatment is particularly beneficial in the prevention of new disease cases while increasing or maintaining the performance of the pigs.
采用双盲随机临床试验,研究tilmicosin (Pulmotil预混料)治疗生长育肥猪临床爆发胸膜肺炎放线杆菌感染的疗效。比较了替尔米科星在饲料中以400 mg/kg的剂量给药和20 mg/kg体重长效土霉素(Terramycine LA)注射治疗临床病猪的效果。两组共147头猪,在给药期15 d和给药后11 d,通过不同的临床和生产性能参数进行比较。用药期间,替米科星组在新发病例数方面具有显著优势(P < 0.01),在切除猪数(P = 0.16)、病猪康复数(P = 0.27)和康复时间(P = 0.42)方面具有不显著优势。在给药后,替米科星组猪的各项临床指标均表现出数值上不显著的改善(P > 0.05)。在整个试验期内(26 d),替尔米霉素组猪的平均日增重和饲料系数均极显著(P < 0.01)优于土霉素组。本研究表明,在饲料中给药400mg /kg的替米考星可有效治疗生长肥育猪胸膜肺炎杆菌感染的临床呼吸道疾病暴发。与临床病猪注射土霉素相比,替尔米科星治疗在提高或维持猪的生产性能的同时,尤其有利于预防新的疾病病例。
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引用次数: 26
Camelid immunoglobulins and their importance for the new-born--a review. 骆驼免疫球蛋白及其对新生儿的重要性综述。
Pub Date : 2001-10-27 DOI: 10.1046/J.1439-0450.2001.00478.X
U. Wernery
Camelid immunoglobulins differ from all other known antibodies and contradict all common theories on antibody diversity. It was demonstrated that up to 75% of all serum proteins are immunoglobulin G (IgG) molecules lacking light chains. IgG2 and IgG3, which only consist of heavy chains, have a low molecular weight which improves their biodistribution and allows a better tissue penetration. Of special importance is the long complementary determining region (CDR) loop which inserts deep into the active site of an enzyme. This binding property was only observed in experiments to gain structural data and to point out the extraordinary value of heavy chain antibodies as biochemical and pharmacological tools. The acquisition and absorption of adequate amounts of colostral immunoglobulins are essential to the health of the neonate. Pre-colostrum serum IgG levels in camelids are low, with concentrations of 0.26 +/- 10.23 mg/ml. Maximum IgG levels are reached after 24 h and kept at a plateau with concentrations of 24.52 +/- 8.8 mg/dl. IgG concentrations above 10 mg/ml indicate a successful passive transfer. IgG levels decline after 2-5 weeks and a marked increase is observed between 1 and 2 months, indicating that the immune system of the neonate has started to mature. A number of different tests are available for the assessment of IgG serum levels. Single radial immunodiffusion (SRID) is the only method that specifically measures serum IgG concentrations. It is a reliable assay to test failure of passive transfer (FPT). FPT is a major factor in neonatal mortality in camelids, but very little has been published so far. Therapeutic administration of colostrum will provide passive protection against infectious diseases for a 2-3-week period of risk, and the intravenous administration of 20-40 ml of camelid plasma helps to combat FPT.
骆驼免疫球蛋白不同于所有其他已知的抗体,与所有关于抗体多样性的常见理论相矛盾。结果表明,高达75%的血清蛋白是缺乏轻链的免疫球蛋白G (IgG)分子。IgG2和IgG3仅由重链组成,具有较低的分子量,这改善了它们的生物分布,并允许更好的组织渗透。特别重要的是插入酶活性位点深处的长互补决定区(CDR)环。这种结合特性仅在实验中观察到,以获得结构数据,并指出重链抗体作为生化和药理工具的非凡价值。获得和吸收足量的初乳免疫球蛋白对新生儿的健康至关重要。骆驼类动物初乳前血清IgG水平较低,浓度为0.26 +/- 10.23 mg/ml。IgG在24 h后达到最大值,并保持在平台浓度为24.52 +/- 8.8 mg/dl。IgG浓度高于10mg /ml表明被动转移成功。2-5周后IgG水平下降,1 - 2个月后IgG水平明显升高,表明新生儿免疫系统开始成熟。有许多不同的测试可用于评估血清IgG水平。单径向免疫扩散(SRID)是唯一一种特异性测定血清IgG浓度的方法。它是检测被动转移(FPT)失败的可靠方法。FPT是骆驼类新生儿死亡的一个主要因素,但迄今为止发表的研究很少。治疗性的初乳管理将提供被动的保护,在2-3周的风险期内防止传染病,静脉注射20-40毫升的骆驼血浆有助于对抗FPT。
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引用次数: 52
Development of a multiplex polymerase chain reaction for the differentiation of bovine herpesvirus-1 and -5. 牛疱疹病毒-1和-5分化的多重聚合酶链反应的建立。
Pub Date : 2001-10-27 DOI: 10.1046/J.1439-0450.2001.00489.X
M. Alegre, M. Nanni, N. Fondevila
Bovine herpesvirus-1 (BHV-1) and bovine herpesvirus-5 (BHV-5) are closely related viruses which exhibit some important differences at the genetic and immunogenic levels which may explain the differences in their pathogenicity and epidemiological characteristics. A multiplex polymerase chain reaction (M-PCR) was developed to detect and differentiate between BHV-1 and BHV-5. In this M-PCR two pairs of primers (TK1, TK2 and GD1, GD2) were used in the same reaction mix to amplify a thymidine kinase genomic region (183 bp) of BHV-1 and one genomic region of the gLycoprotein D (564 bp) of BHV-5. The specificity of the M-PCR was demonstrated when using both primers pairs simultaneously with BHV-1 and BHV-5 templates. The two expected bands were amplified without the apparition of non-specific products. However, when other herpesvirus strains were used, there was no amplification. To evaluate the sensitivity of the assay, dilutions of purified viral DNA were made for M-PCR amplification. The detection limit was 7 pg for BHV-1 and 22 pg for BHV-5. It was also determined by comparing the M-PCR with viral isolation. M-PCR was able to detect one log10 more than viral isolation for BHV-1 and for BHV-5 was two logarithms lower. The applicability of M-PCR was demonstrated on different specimens. Twenty isolates from field samples (11 BHV-1 and nine BHV-5) were positive by M-PCR, and the results were completely coincident with previous characterization using the immunoperoxidase assay. M-PCR could detect viral DNA in organ samples from natural infections, such as semen and brain. In addition, M-PCR detected more positive samples than observation of the citophatic effect in cell culture of nasal swabs from experimentally infected animals in two different assays. Owing to the difference in size of the M-PCR products which allows easy identification in an electrophoretic run, it is not necessary to use extra blotting and hybridization steps or a second round of amplification to differentiate clearly between BHV-1 and BHV-5.
牛疱疹病毒-1 (BHV-1)和牛疱疹病毒-5 (BHV-5)是亲缘关系密切的病毒,在遗传和免疫原水平上表现出一些重要的差异,这可以解释它们在致病性和流行病学特征上的差异。建立了多重聚合酶链反应(M-PCR)检测和区分BHV-1和BHV-5。在M-PCR中,使用两对引物(TK1、TK2和GD1、GD2)在相同的反应混合物中扩增BHV-1的胸苷激酶基因组区域(183 bp)和BHV-5的糖蛋白D基因组区域(564 bp)。当两对引物同时用于BHV-1和BHV-5模板时,M-PCR的特异性得到了验证。在没有非特异性产物出现的情况下,这两个预期的条带被扩增。然而,当使用其他疱疹病毒株时,没有扩增。为了评估检测的敏感性,将纯化的病毒DNA进行稀释,进行M-PCR扩增。BHV-1的检出限为7 pg, BHV-5的检出限为22 pg。并将M-PCR与病毒分离进行比较。M-PCR检测BHV-1比病毒分离多1个对数,BHV-5比分离低2个对数。M-PCR在不同标本上的适用性得到了验证。从田间样品中分离得到的20株BHV-1(11株)和BHV-5(9株)经M-PCR检测呈阳性,结果与先前用免疫过氧化物酶法鉴定的结果完全一致。M-PCR可以检测自然感染器官样本中的病毒DNA,比如精液和大脑。此外,在两种不同的检测中,M-PCR检测到的阳性样本比实验感染动物鼻拭子细胞培养中观察到的citophatic效应更多。由于M-PCR产物的大小不同,这使得在电泳运行中易于识别,因此不需要使用额外的印迹和杂交步骤或第二轮扩增来明确区分BHV-1和BHV-5。
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引用次数: 54
Products of fourth-stage larvae of Oesophagostomum dentatum induce proliferation in naïve porcine mononuclear cells. 食道齿状口第四期幼虫产物诱导naïve猪单核细胞增殖。
Pub Date : 2001-10-27 DOI: 10.1046/J.1439-0450.2001.00483.X
R. Freigofas, W. Leibold, A. Daugschies, A. Joachim, H. Schuberth
Infection of pigs with Oesophagostomum dentatum is a major cause of economic losses in pig productions. Whether infection with this nematode results in a protective immunity is still in debate and information about immune-modulating properties of O. dentatum are lacking. The present study investigated the question whether products of O. dentatum larvae modulate the proliferative response of porcine blood mononuclear cells (poMNC) in vitro. The poMNC of naïve and O. dentatum-infected pigs were cultured for 72 h in the presence of products (total homogenates and culture supernates) derived from third- (L3) and fourth-stage larvae (L4) of O. dentatum. Numbers of vital cells and blast-transformed cells were determined flow cytometrically. No larvae product induced an accelerated death of poMNC in vitro. In contrast, products of L4 (but not L3) significantly increased the numbers of vital poMNC in vitro (up to 187%). In addition, L4 products (homogenates and supernates, 0.1-10 microg/ml) but not those of L3 induced significant blastogenesis of poMNC. This was seen with poMNC from naïve and from O. dentatum-infected animals. In spite of these effects, the larvae products were not able to modulate the mitogen-induced (Concanavalin A) poMNC proliferation of naïve and infected animals. In summary, larvae of O. dentatum contain and secrete products with potential immunomodulatory capacity for porcine peripheral blood mononuclear cells. The differential effects of L3 and indicate that the parasite alters its set immunomodulatory substances during its development. This has to be considered in further studies and may help to identify the mediators involved.
猪食道齿状口感染是猪生产经济损失的主要原因。感染这种线虫是否会导致保护性免疫仍然存在争议,并且缺乏关于齿状线虫免疫调节特性的信息。本研究探讨了齿状棘球蚴幼虫产物是否能调节猪血单核细胞(poMNC)的体外增殖反应。将naïve和O. dentatum感染猪的poMNC在O. dentatum第三期(L3)和第四期幼虫(L4)的产品(总匀浆和培养上清)中培养72 h。流式细胞术检测存活细胞和胚转化细胞的数量。无幼虫产物诱导poMNC体外加速死亡。相比之下,L4的产物(而不是L3)显著增加了体外重要poMNC的数量(高达187%)。此外,L4的产物(匀浆和上清,0.1 ~ 10 μ g/ml)能诱导poMNC显著的胚发生,而L3的产物则不能。这在naïve的poMNC和感染齿形o的动物身上可见。尽管有这些影响,但幼虫产物不能调节naïve和感染动物的丝裂原诱导的poMNC增殖。综上所述,齿状棘球蚴幼虫含有并分泌对猪外周血单核细胞具有潜在免疫调节能力的产物。L3和的差异作用表明寄生虫在其发育过程中改变了其免疫调节物质。在进一步的研究中必须考虑到这一点,并可能有助于确定所涉及的介质。
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引用次数: 7
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Journal of veterinary medicine. B, Infectious diseases and veterinary public health
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