隐孢子虫和贾第鞭毛虫双重实时荧光PCR检测方法的建立

本研究根据隐孢子虫和贾第鞭毛虫相对保守序列,设计2对引物及其相应的Taqman探针,通过对引物和探针浓度、Taq酶以及反应条件等优化筛选后,建立了能同时检测隐孢子虫和贾第鞭毛虫的双重实时荧光PCR方法。所建立的隐孢子虫和贾第鞭毛虫双重荧光PCR检测方法的灵敏度为10-9,相当于46.1copies/!L,重复性好,特异性佳。本研究建立的双重荧光PCR技术,可以快速、准确地于一管一次反应检测隐孢子虫和贾第鞭毛虫。     

宁夏部分地区奶牛贾第鞭毛虫分子流行病学调查

为了明确宁夏地区奶牛贾第鞭毛虫的优势基因型及其流行特征，试验首先于宁夏地区10个规模化奶牛养殖场采集880份奶牛粪便样品，针对贾第鞭毛虫保守基因——β-贾第素(β-giardin, βg)基因序列设计引物，然后以提取的粪便样品全基因组DNA为模板进行巢式PCR,将PCR产物的测序结果进行BLAST比对分析，鉴定贾第鞭毛虫的基因型，并建立系统进化树，最后对宁夏地区不同城市、不同月龄及腹泻和非腹泻奶牛粪便样品的感染阳性率和基因型进行统计。结果表明：880份粪便样品中，有145份样品扩增到大小为510 bp的条带，贾第鞭毛虫感染阳性率为16.5%(145/880),其中集聚体A的占比为1.4%(2/145),集聚体E的占比为98.6%(143/145);所有集聚体A的βg基因为1种序列(已上传至GenBank,登录号为OQ101261 1NX),所有集聚体E的βg基因为4种序列(已上传至GenBank,登录号分别为OQ101262 2NX、OQ101263 3NX、OQ101264 4NX和OQ101265 5NX);OQ101261 1NX(奶牛)与MK5773339.1(藏牛)、MF49...     

贾第鞭毛虫(Giardia)对犊牛的实验感染

<正> 贾第鞭毛虫(Giardia)是一种分布广泛,可引起腹泻和痢疾的常见寄生性原虫。它寄生于脊椎动物小肠(尤其十二指肠)和大肠内,宿主范围广,包括人、牛、犬、猫、鼠、马等。不同宿主的贾第鞭毛虫形态学特征相似,所以Filia(1952)指出哺乳动物中只有两个种,即十二指肠贾第鞭毛虫(G.duodenalis)和鼠贾第鞭毛虫(G.muris)。但目前都用不同宿主命名,如犬贾第鞭毛虫(G.Canis)、牛贾第鞭毛虫(G.bovis)、蓝氏贾第鞭毛虫(G.Lambia)、猫贾第鞭毛虫(G.Cati)等等(Levine 1985)。交互感染试验显示了贾第鞭毛虫人畜共患的特点,Visvesvara(1980)发现从牛、豚鼠和人得到的贾第鞭毛虫其抗原性相同,并指出哺乳动物,除     

牦牛粪样中蓝氏贾第鞭毛虫巢式PCR检测方法的建立

采用建立的巢式PCR方法检测了297份牦牛粪样。结果表明:建立的巢式PCR方法一扩的最佳退火温度为55℃,二扩的最佳退火温度为53℃;对牦牛源贾第鞭毛虫的最低检测量为1个虫体DNA/m L;特异性试验表明,对隐孢子虫、弓形虫、柔嫩艾美尔球虫、阿米巴虫、牛巴贝斯虫等寄生虫DNA的扩增均为阴性;297份牦牛粪样中贾第鞭毛虫的检出率为7.4%。表明建立的方法具有较高的敏感性和特异性,可用于牦牛源贾第鞭毛虫感染的检测。     

蓝氏贾第鞭毛虫检测技术研究进展

蓝氏贾第鞭毛虫是一种分布广、危害严重的水源性人兽共患的寄生虫原虫,可致人和动物严重腹泻.由于其种类繁多,易传染,传播快,对公共卫生安全造成严重威胁.传统的贾第鞭毛虫检测方法主要为粪便样本的显微镜检查.随着免疫学和现代分子生物学的不断发展,很多分子生物学检测技术不断涌现,并成功应用到对贾第鞭毛虫的快速检测.论文就近年来贾第鞭毛虫的检测技术进展情况进行综述.     

牛源蓝氏贾第虫环介导等温扩增检测方法的建立及应用

为建立一种适用于牛场或基层单位的牛源蓝氏贾第虫快速检测方法,根据NCBI网站(https://www.ncbi.nlm.nih.gov/)上的蓝氏贾第虫基因序列设计外引物(FIP,BIP)、内引物(F3,B3)和环引物(LB),建立了牛源蓝氏贾第虫的环介导等温扩增(loop-mediated isothermal amplification, LAMP)检测方法,并对采集的40份犊牛新鲜粪便样品分别采用巢式PCR方法(贾第虫检测金标准)和建立的LAMP检测方法进行蓝氏贾第虫检测。结果显示：建立的牛源蓝氏贾第虫LAMP检测方法特异性良好,灵敏度高于巢式PCR;2种方法检测结果的符合率为100%。结果表明：所建立的LAMP检测方法可用于农场或基层单位的牛源蓝氏贾第虫检测。     

猫贾第鞭毛虫感染的诊治

贾第鞭毛虫病(Giardiasis)是由贾第鞭毛虫(Giardia,简称贾第虫)引起的一种以腹泻为主要症状的肠道疾患。贾第鞭毛虫是世界各地人类和绝大多数动物的肠道内原虫。起初贾第鞭毛虫仅被视为一种共生性的肠道原虫,自1976年以来,由于世界各地相继发生本病,甚至暴发流行,人们才真正认识其致病性。     

三江源地区饮用水源中隐孢子虫和贾地鞭毛虫的检测

通过膜过滤、絮凝沉淀法浓缩收集了三江源地区水源中隐孢子虫和贾第鞭毛虫,并用蔗糖漂浮法纯化了上述"两虫"。及抗酸染色后镜检,结果显示:在三江源地区10条河流中的6条(60%)检出隐孢子虫,32个采样点11个点(34%)的样品检出隐孢子虫;在10条河流中有2条(20%)检出贾地鞭毛虫,32个采样点的4个点(12.5%)的样品中检出贾地鞭毛虫。     

球虫和其他原虫(续完)

<正>5重要性较小或有可能传播给人类的其他原虫5.1贾第鞭毛虫蓝氏贾第鞭毛虫(Giardia duodenalis),也叫做蓝伯氏贾第虫(Giardia lamblia)和肠贾第虫(Giardia intestinalis),可能是世界各地最常见的人和家畜的肠道寄生虫。它是一个物种群,根据基因分析有7个基因集合体,它们的成员在形态学上不能区别,集合体A和B可感染人和多种哺乳动物;其他的主要是感染动物的特定群,如集合体C和D可感染犬科动物,集合体E可     

青海牦牛源贾第虫的鉴定及分子特征分析

从青海省祁连县采集93份牦牛粪样用于贾第虫检测。所有样品经蔗糖密度梯度离心法纯化处理后,采用卢戈氏碘液染色镜检和免疫荧光试验进行检测,将镜检阳性样品采用套式PCR方法扩增18S rRNA,并对扩增产物进行测序分析。结果镜检和免疫荧光试验中有9份样品为贾第虫阳性,阳性率为9.7%。镜检阳性样品采用套式PCR方法扩增后有8份样品为18SrRNA基因阳性,产物长度292bp,测序后的序列分别命名为QHQL201501~QHQL201508。系统发育分析显示,分离的虫株属于牛源十二指肠贾第虫,基因型均为反刍动物特有的聚集体E,未发现具有人兽共患潜力的A型和B型。     

Comparison of in vitro and in situ methods in evaluation of forage digestibility in ruminants

The objective of this study was to compare the application of different in vitro and in situ methods in empirical and mechanistic predictions of in vivo OM digestibility (OMD) and their associations to near-infrared reflectance spectroscopy spectra for a variety of forages. Apparent in vivo OMD of silages made from alfalfa (n = 2), corn (n = 9), corn stover (n = 2), grass (n = 11), whole crops of wheat and barley (n = 8) and red clover (n = 7), and fresh alfalfa (n = 1), grass hays (n = 5), and wheat straws (n = 5) had previously been determined in sheep. Concentrations of indigestible NDF (iNDF) in all forage samples were determined by a 288-h ruminal in situ incubation. Gas production of isolated forage NDF was measured by in vitro incubations for 72 h. In vitro pepsin-cellulase OM solubility (OMS) of the forages was determined by a 2-step gravimetric digestion method. Samples were also subjected to a 2-step determination of in vitro OMD based on buffered rumen fluid and pepsin. Further, rumen fluid digestible OM was determined from a single 96-h incubation at 38°C. Digestibility of OM from the in situ and the in vitro incubations was calculated according to published empirical equations, which were either forage specific or general (1 equation for all forages) within method. Indigestible NDF was also used in a mechanistic model to predict OMD. Predictions of OMD were evaluated by residual analysis using the GLM procedure in SAS. In vitro OMS in a general prediction equation of OMD did not display a significant forage-type effect on the residuals (observed - predicted OMD; P = 0.10). Predictions of OMD within forage types were consistent between iNDF and the 2-step in vitro method based on rumen fluid. Root mean square error of OMD was least (0.032) when the prediction was based on a general forage equation of OMS. However, regenerating a simple regression for iNDF by omitting alfalfa and wheat straw reduced the root mean square error of OMD to 0.025. Indigestible NDF in a general forage equation predicted OMD without any bias (P ≥ 0.16), and root mean square error of prediction was smallest among all methods when alfalfa and wheat straw samples were excluded. Our study suggests that compared with the in vitro laboratory methods, iNDF used in forage-specific equations will improve overall predictions of forage in vivo OMD. The in vitro and in situ methods performed equally well in calibrations of iNDF or OMD by near-infrared reflectance spectroscopy.     

大庆市羊东毕吸虫病流行病学调查

对大庆市羊东毕吸虫病的流行病学进行了调查研究。结果表明，大庆有3种东毕吸虫，即程氏东毕吸虫（O.cheni)、土耳其斯坦东毕吸虫（O.turkestanica)和土耳斯坦东毕吸虫结节变种（O.turkestanican var.thuberculata)；中间宿主螺类有1种，即卵萝卜螺；东毕吸虫病在大庆分布很广，所调查的4区2县均有发生，羊东毕吸虫平均感染率分别为50．33％，平均死亡率为21．12％，大庆羊急性病例发病时间为8月上旬至10月下旬，慢性病例为11月上旬至翌年2月中旬。     

ELISA试验在防制猪伪狂犬病中的应用

猪伪狂犬病是由伪狂病病毒(Pseudorabies virus PRV)引起的急性传染病。其特征是成年猪常为隐性感染，可有流产、死胎、呼吸系统症状，新生仔猪除神经症状外还可侵害消化系统，并伴大批死亡。本病已经给全世界的养猪业造成严重的损失，我国许多地区已报道该病。为进一步探讨该病的控制、净化措施，我们对12个养猪场共856分血清采用美国嗍公司的ELISA试剂盒进行了抗体检测。     

中国圈养野生动物疫苗使用调查

2005年对中国动物园协会单位中的18家动物园饲养野生动物疫苗免疫情况进行调查。受调查动物园在野生动物疫苗使用和动物种类方面具有一定的代表性。调查结果显示动物园动物现共使用18类36种疫苗,预防31种疫病。其中哺乳动物使用14类24种疫苗,预防24种疫病;禽类使用4类12种疫苗,预防7种疫病。使用范围最广的有禽流感、犬瘟热疫苗等、新城疫疫苗、猫瘟热疫苗等。共有24目58科动物接种疫苗,其中食肉动物类1目8科,食草动物类3目9科,杂食动物类2目3科,禽类18目38科。研究结果提示应加强动物园之间疫病信息交流、防疫资源的利用、疫病监测和研究,加大对动物园动物疫病的研究投入,逐步建立动物园动物统一的防疫规程。     

Estimate of economic impact of fowl cholera in turkeys in Georgia in 1986

Information gathered from cases of fowl cholera (FC) in commercial turkey flocks through case records, flock records, and telephone and mail surveys was used to estimate disease costs. The cost to the Georgia commercial turkey industry in 1986 from preventive measures, treatment of outbreaks, and production losses from the disease was estimated at $634,545. The cost of FC per kg of live production was estimated to be $0.015.     

Use of cyclocryotherapy in management of glaucoma in dogs

In 5 cases of glaucoma (2 from trauma, 2 from narrowed drainage angles, 1 secondary to lens extraction), cyclocryotherapy was used to control intraocular pressure. In all cases the intraocular pressure decreased, with the usual result being a cosmetic and painless but blind eye.     

Occurrence of MRSA in pigs and in humans involved in pig production--preliminary results of a study in the northwest of Germany

In 2007, 678 pigs of all age groups out of 347 different farms from Lower Saxony and Northrhine-Westphalia and 86 persons occupationally exposed to pigs were investigated for their nasal colonisation with methicillin-resistant Staphylococcus areus (MRSA) by the Field Station for Epidemiology of the University of Veterinary Medicine Hannover and the Robert Koch-Institute. At the individual animal level, a frequency of positive results of 13% (n = 85 positive animals) and at the herd level, a frequency of positive results of 18% (n = 62 positive herds) were found. All isolates were assigned to the Multilocus Sequence Typing Type ST398. Within MRSA-positive herds, there were more MRSA-negative than MRSA-positive animals. Among the occupationally exposed persons (veterinarians, laboratory personnel and meat inspection personnel), 20 persons (23%) showed a nasal colonisation with MRSA ST398. A quite strong association between the intensity of the contact to pigs with the frequency of nasal colonisation in the occupationally exposed persons was detected. None of the animals or the humans nasally colonised by MRSA ST398 showed any clinical symptoms of a staphylococcal infection. Conclusions are drawn on the herd and intra-herd prevalence of the nasal colonisation of pigs with MRSA ST398 in pigs, but especially on which questions need to be addressed by further research.     

2017年西藏拉萨市家禽禽流感抗体水平检测

为了解西藏拉萨市家禽禽流感抗体水平,2017年8月随机选择免疫过H5禽流感疫苗的8个规模养殖场、2个散养户、3个活禽交易市场,共13个群体,采集1 041份家禽血清样品进行H5、H7、H9亚型禽流感HI抗体检测。结果显示：H5亚型禽流感免疫抗体合格率为45.73%,H7、H9亚型抗体阳性率分别为21.13%、52.64%;养殖场、散养户和活禽交易市场的不同亚型抗体水平存在较大差异,规模化养殖场的H7亚型抗体阳性率最高（29.92%）,活禽市场的H5、H9亚型阳性率最高（78.16%、79.31%）,散养户的所有亚型抗体阳性率均最低。结果表明,拉萨市H5亚型禽流感疫苗免疫效果较差,特别是散养户,疫病发生风险较高,H7、H9亚型疑似自然感染率较高,应进一步查明原因,及时采取相应的措施,防止疫情发生。