内蒙古锡林郭勒地区牛羊莱姆病的流行病学调查

本项研究于2000年5月至2001年5月从内蒙古锡林郭勒盟地区，不同草原景观带牛、羊体表和草地采集草原血蜱1800只。用直接荧光接体法和暗视野显微镜检查蜱带菌情况，带菌率分别为7．5％（6／80）和2％（2／100）。用间接荧光法进行血清流行病学调查，结果证实，调查点的牛和野鼠均存在莱姆病螺旋体感染，其感染率分别为10％和20％，都高于当地人群平均感染率（7．09％）。上述研究证实内蒙古锡林郭勒地区人畜及野生动物均有莱姆病螺旋体感染。     

青海省部分地区动物莱姆病流行病学检测

采用间接免疫荧光抗体法进行牛、鼠血清流行病学调查检测,用直接荧光抗体法和暗视野显微镜检查蜱虫带菌情况。结果表明牛莱姆病平均感染率12.45%,羊莱姆病平均感染率9.37%,蜱虫平均带菌率25.29%,鼠类感染率17.2%。证实了青海省部分地区动物、昆虫体内携带莱姆病伯氏疏螺旋体。     

柴达木盆地都兰县多种家畜感染莱姆病及媒介昆虫带菌调查

调查都兰县各种动物感染莱姆病情况。用间接免疫荧光抗体法(IFA)对牦牛、绵羊、骆驼等家畜进行血清流行病学调查,用直接荧光抗体法和暗视野显微镜检查蜱虫带菌情况。结果:黄牛血清20份,阳性3份。阳性率15%;绵羊血清20份,阳性1份,阳性率5%;骆驼血清25份,阳性10份,阳性率40%;草原革蜱203只,阳性7只,阳性率3.44%;加瓦尔血蜱74只,阳性2只,阳性率2.7%;结论:处于柴达木盆地的都兰县是莱姆病自然疫源地。     

伯氏疏螺旋体套式PCR检测方法建立及应用

为了解新疆北疆地区硬蜱携带莱姆病病原伯氏疏螺旋体的流行现状,选择莱姆病病原伯氏疏螺旋体5S-23S rRNA基因间隔区设计嵌套式引物,建立了具有较高敏感性和特异性的伯氏疏螺旋体套式PCR检测方法,应用该方法对2018—2019年在新疆北疆6个地区采集的硬蜱进行伯氏疏螺旋体带菌率检测。结果显示:伯氏疏螺旋体总阳性率为15.17%(81/534),其中青河县阳性率最高,达23.64%(26/110)。提示:应进一步加强及重视北疆地区莱姆病的防控工作。     

采用PCR方法对我国蜱伯氏疏螺旋体感染的流行病学调查

通过调查我国已有的4种蜱伯氏疏螺旋体的感染情况,以进一步阐明莱姆病在中国的分布与流行状况。本研究通过筛选扩增伯氏疏螺旋体外膜蛋白A(OspA)基因片段的通用引物,获得1对特异性引物,优化反应条件后建立用于检测蜱体内伯氏疏螺旋体的PCR方法,其扩增片段大小为307 bp。该方法可检测出10 pg的阿氏疏螺旋体(Ba)、1 pg的伽氏疏螺旋体(Bg)和0.01 pg的狭义伯氏疏螺旋体(Bb)的3种不同基因型的伯氏疏螺旋体的OspA基因组DNA,表明其敏感性较好,适用于蜱感染伯氏疏螺旋体状况的调查。本研究从我国7个省采集到的667只蜱,进行伯氏疏螺旋体感染的流行病学调查,分类鉴定表明这些蜱分属革蜱属、血蜱属、牛蜱属和扇头蜱属。PCR检测所获数据表明它们的感染率分别为4%(10/264)、6%(11/137)、31.4%(59/185)和31%     

荧光定量PCR检测硬蜱体内莱姆病螺旋体的研究

为建立一个荧光定量PCR检测硬蜱体内莱姆病螺旋体的方法,根据GenBank登录的莱姆病螺旋体鞭毛蛋白FlaB序列,应用生物学软件进行序列比对,在保守的C段区设计与筛选特异引物和TaqMan探针。对荧光定量PCR反应体系与条件进行优化,验证方法的特异性、敏感性,并通过对感染螺旋体的蜱样本的检测,评价该方法的实用价值。结果显示,自然感染莱姆病螺旋体的35份蜱标本检测阳性符合率100%,正常蜱20份标本的检测结果均为阴性。该方法对牛巴贝斯原虫、泰勒原虫、边缘无浆体、金龟子绿僵菌、大肠杆菌等蜱体常见病原微生物所抽提的DNA的检测均呈阴性。荧光定量PCR方法检测质粒的灵敏度可达1×102拷贝/μL。TaqMan荧光定量PCR方法检测硬蜱体内莱姆病螺旋体具有较好的敏感性和特异性,可适于莱姆病的流行病学调查和监控。     

国内莱姆病研究进展

莱姆病是由伯氏疏螺旋体引起的,由蜱传播的自然疫源性疾病,是新发现的人兽共患传染病,主要经蜱叮咬人、兽而传播。该病分布广泛,全世界至今已有五大洲的70多个国家报告发现了该病,而且发病区域仍在继续扩大,发病率呈现上升趋势。我国自从1986年调查发现东北林区人群中存在莱姆病的发生和流行以来,现已证实29个省(市、区)的人群中存在莱姆病的感染,并从病原学上证实其中至少有19个省(市、区)存在该病的自然疫源地,人群中有莱姆病的发生和流行。对动物宿主的调查证实,牛、马、羊、狗、鼠等均存在莱姆病感染,不同地区及不同种动物莱姆病感染存在较大差异。论文从莱姆病的病原分类、流行病学、传播途径、传染源和分子生物学等几方面阐述了莱姆病在中国的流行情况,为进一步了解莱姆病的流行及有效防控提供参考。     

吸血蓖子硬蜱成虫唾腺莱姆病螺旋体重度感染的发现

为了解莱姆病螺旋体在蜱体内的发育生物学，采用直接免疫荧光试验、银染色法组织学及电子显微镜观察螺旋体，结果显示，感染莱姆病螺旋体的饥饿蓖子硬蜱雌虫未吸血与吸血0．5和1天唾腺中无螺旋体。在吸血开始后2天出现唾腺的轻度感染，3天出现唾腺的中度感染，4天和5天出现重度感染。这些结果表明，莱姆病螺旋体媒介蜱唾腺的感染程度随吸血时间的延长而迅速增高，并可在缓慢吸血期的中后期形成重度感染。莱姆病螺旋体在吸血期媒介唾腺内的大量存在，特别是重度感染的发现，为该病原螺旋体由蟀向脊椎动物宿主传播的唾液分泌途径假设提供了有力证据。本研究对莱姆病螺旋体在吸血蟀唾腺内分裂增殖的可能性进行了讨论。     

青海部分地区牛羊蜱等外寄生虫调查

为了了解青海部分地区牛羊外寄生虫的感染情况,我们用随机抽样法对海晏等8县(市)的55头牛和119只羊进行了调查,结果发现6种外寄生虫,属于2纲3目4科4属,感染率分别为草原革蜱21.43%,牛颚虱15.80%,牛皮蝇(蛆)57.14%;羊的草原革蜱44.25%,羊蜱蝇46.22%,足颚虱37.5%,绵羊颚虱16.07%。     

东北地区蜱携带的肝原虫分子检测

为检测东北地区蜱传肝原虫,于2015年4—6月通过布旗法在吉林省和黑龙江省采集蜱2 767只,其中森林革蜱168只、草原革蜱212只、全沟硬蜱1 629只、嗜群血蜱378只、长角血蜱380只。根据蜱种和地区分组,每组约15只,提取基因组DNA,应用巢式PCR方法检测肝原虫18S rRNA,阳性样本克隆、测序并进行系统进化分析,利用MLE(Maximum Likelihood Estimation)法计算感染率。东北地区蜱的肝原虫感染率为1.6%(43/189),森林革蜱、草原革蜱、全沟硬蜱、嗜群血蜱和长角血蜱的肝原虫感染率分别为2.8%(4/12)、1.5%(3/16)、2.0%(29/110)、0.5%(2/26)和1.1%(5/25),吉林省和黑龙江省肝原虫感染率最高的蜱种分别为森林革蜱(2.8%)和全沟硬蜱(2.1%),进化分析结果显示东北地区蜱携带的肝原虫为2个不同基因型,且均与日本貂(Martes melampus melampus)肝原虫的同源性为99%,均能在草原革蜱和全沟硬蜱中检出,而且在不同地区蜱的感染率存在一定差异。本研究确定东北地区存在蜱感染肝原虫,但是其分类地位及动物宿主还需进一步鉴定。     

西门塔尔牛生产特性分析与高效养殖技术措施

西门塔尔牛为乳肉兼用品种。是世界第二大牛品种,全世界现有存栏数为4100多万头。内蒙古是中国西门塔尔牛品种培育基地,西门塔尔牛已成为该地区肉牛产业的主导品种。2013年末,内蒙古肉牛存栏570万头．其中90％以上的良种与改良种为西f1塔尔血统。在对西门塔尔牛生产特性进行详细分析的基础上,提出了养好西门塔尔牛的技术措施,以期为内蒙古地区的西门塔尔牛养殖提供参考。     

Evaluation of different media and a BGM cell culture assay for isolation of Borrelia burgdorferi sensu lato from ticks and dogs

A co-culture assay for isolation of Borrelia burgdorferi sensu lato (sl.) from naturally infected ticks and dogs suspected of Lyme borreliosis (LB) was evaluated using buffalo-green-monkey (BGM) cells as the mammalian component. Four different media were tested for their ability to provide sufficient growth conditions for spirochetes and BGM cells. A total of 176 Ixodes ricinus ticks and 268 specimens from 98 dogs were used to compare cell-free culture with the BGM co-culture. A 1:1 mixture of Barbour-Stoenner-Kelly medium (BSK) and Eagle's minimum essential medium (EMEM) supported the growth of the two test strains, B. burgdorferi sensu stricto B31 and B. valaisiana VS116 to the same extent as BSK medium and the growth as well as the viability of BGM cells in this medium were the same as in EMEM. Using the 1:1 mixture of BSK and EMEM, borrelial growth measured in co-culture with BGM cells did not differ significantly from corresponding values obtained in cell-free cultures. In cell-free culture the isolation rate of B. burgdorferi sl. from ticks was significantly higher in BSK/EMEM 1:1 than in BSK medium (P < 0.01). Co-culture with BGM cells had no significant influence on the isolation rate of borreliae from ticks. However, a significant amount of isolates were obtained by one of the procedures only. Analysing canine specimens accordingly, spirochetes were grown from the blood of one dog after four weeks in BGM cell co-culture. The isolate was classified as B. afzelii by PCR-coupled restriction fragment length polymorphism analysis.     

北方非传统肉牛养殖区品种改良方案

在目前全国各地肉牛产业飞速发展的形势下,北方非传统肉牛养殖区也开始将产业重点向肉牛养殖业调整。对北方非传统肉牛养殖区的生态环境、肉牛品种、饲养管理、产业发展和肉牛交易等特点进行了分析。在该基础上,因地制宜地提出北方非传统肉牛养殖区肉牛品种改良方案。该方案分为2个阶段实施,第1阶段为基础母牛安格斯化：利用红安格斯牛作为父本,对当地黄牛进行杂交改良,实现基础母牛群体安格斯化;第2阶段为两品种轮回杂交：选用另一专门化肉牛品种与红安格斯牛为主要父本品种,开展两品种轮回杂交,建立优质肉牛高效繁育技术体系。通过制定该技术方案,以期为北方非传统肉牛养殖区开展黄牛改良和发展特色经济提供技术指导。     

Molecular detection of Anaplasma phagocytophilum in cattle and Ixodes persulcatus ticks

The tick-borne pathogen, Anaplasma phagocytophilum (A. phagocytophilum), the causative agent of human granulocytic anaplasmosis (HGA), is increasingly becoming a public health concern as an aetiological agent for emerging infectious disease. We found A. phagocytophilum infection in a pooled sample of field-collected Ixodes persulcatus (I. persulcatus) ticks from one district in Hokkaido, Japan. Thus, to further investigate the prevalence in field-collected ticks, we used PCR assays targeting the A. phagocytophilum gene encoding 44 kDa major outer membrane protein (p44) for screening of I. persulcatus ticks and samples from cattle from pastures. Out of the 281 I. persulcatus ticks, 20 (7.1%) were found to harbor A. phagocytophilum DNA. The infection rate for A. phagocytophilum in cattle was 3.4% (42/1251). In future studies, it will be necessary to investigate effects of the infection in order to understand its pathogenesis of A. phagocytophilum in domestic animals.     

内蒙古部分地区放牧牛羊弓形虫血清学调查

[目的]了解内蒙古部分地区放牧牛羊弓形虫病感染情况。[方法]采用间接血凝试验(IHA)对阿拉善盟及呼伦贝尔市随机采集的286份放牧牛羊血清样本进行弓形虫抗体检测。[结果]绵羊血清弓形虫抗体总阳性率为1.68%,其中,阿拉善盟阳性率为4.84%,呼伦贝尔市阳性率为0.56%;经统计学分析,两个地区的绵羊血清弓形虫抗体阳性率差异不显著(P>0.05);牛血清弓形虫抗体总阳性率为0。[结论]内蒙古主要牧区放牧绵羊存在弓形虫感染,应引起足够的重视,并制定相应的弓形虫病防治措施。     

Study on Supplementary Quantity of Grazing Beef Cattle in Spring in Inner Mongolia Grassland

The experiment was conducted to study the optimal supplementary feeding quantity of grazing beef cattle in spring in Inner Mongolia grassland.Seventy-five Simmental beef cattle with average weight of (281.19±17.15)kg were selected and divided into 5 groups (group A,B,C,D and E) with 15 replicates per group.Those cattle grazed optionally and supplemented with 0,0.8,1.7,2.5,3.4 kg/d concentrate,respectively,according to beef cattle weight of 0,0.3%,0.6%,0.9% and 1.2%.Pre-trial period was 15 d,the trial period was 60 d.The results showed that,with the increasing of concentrate supplementation,the daily gain of beef cattle was gradually significant increased (P < 0.05);Group D were 331.25%,130.00% and 38.00% higher than that of group A,B and C,respectively,the apparent digestibility of dry matter and crude protein were significantly higher than that of other groups (P < 0.05).The daily income of group D was 11.22 yuan and were 183.33%,102.53%,36.17%,20.52% significantly higher than that of group A,B,C and E (P < 0.05),the fattening income of group D was the highest.According to above results,the optimum adding levels was 2.5 kg/d in spring under the condition of grazing in Inner Mongolia.The higher daily gain and higher economic benefits could be obtained under the optimal supplementary feeding quantity.     

内蒙古草原放牧肉牛春季补饲量的研究

试验旨在研究内蒙古草原放牧肉牛春季最佳补饲量。选择75头体重为(281.19±17.15)kg的健康西门塔尔杂交肉牛随机分为5组(A、B、C、D、E组),每组15头牛,1头牛为1个重复。各组分别按体重的0、0.3%、0.6%、0.9%及1.2%补饲精料,依次为0、0.8、1.7、2.5及3.4 kg/d,其他放牧条件相同。预试期15 d,正试期60 d。结果表明,随着精料补饲量的增加,肉牛的平均日增重依次显著增加(P < 0.05),D组平均日增重较A、B、C组分别提高331.25%、130.00%及38.00%,日粮中干物质和粗蛋白质的表观消化率显著高于其他各组(P < 0.05);D组日均收入为11.22元,分别较A、B、C及E组显著提高了183.33%、102.53%、36.17%及20.52%(P < 0.05),育肥效益最好。综合以上结果,内蒙古草原春季放牧条件下,肉牛每天的最佳补饲量为2.5 kg,在此条件下可获得较高的日增重及较高的经济收益。     

济宁青山羊BMP15基因外显子2的克隆与序列分析

本研究对高繁殖力的山羊品种济宁青山羊和低繁殖力的山羊品种内蒙古绒山羊共20只母羊的骨形态发生蛋白15（bone morphogenetic protein 15, BMP15）基因第2外显子的扩增产物进行了克隆测序及序列比较分析。结果表明：BMP15基因第2外显子的第3对引物扩增片段存在多态。济宁青山羊与内蒙古绒山羊相比，其BMP15基因外显子2差异由2个核苷酸和2个氨基酸改变组成，核苷酸同源性为99%。济宁青山羊与鸡、小鼠、人、猪、牛、绵羊之间BMP15基因外显子2的核苷酸序列同源性为71%～99%，氨基酸序列同源性为43%～99%。     

The first survey of Theileria orientalis infection in Mongolian cattle

In the present study, we have surveyed the presence of a bovine Theileria protozoan, Theileria orientalis, in Mongolian cattle and engorging tick populations from selected provinces and districts in Mongolia. The percentages of infection in the cattle and ticks ranged from 8.8 to 66.6 and from 3.7 to 73.3, respectively, on a per district basis. The genetic diversity of T. orientalis isolates was also studied, based on the protozoan gene encoding a major piroplasm surface protein (MPSP). At least five genotypes (types 1, 3, 5, 7, and N-3) of T. orientalis were found to be circulating among the Mongolian cattle and tick populations. In particular, types 3 and N-3 were common in most of the districts examined, while a strong geographical relationship among the genotypes was not detected in the present study. This is the first epidemiological report describing the presence of T. orientalis infection in Mongolian cattle.