人和动物丝虫共生菌——沃尔巴克氏体研究进展

沃尔巴克氏体(Wolbachia,Wb)是在丝虫体内发现的一类共生菌,它诱导的免疫病理引起的炎症反应是丝虫感染后致病的主要原因之一。论文论述了丝虫沃尔巴克氏体的分类、结构与分布、生殖及传播方式、基因组和沃尔巴克氏体与丝虫之间的相互作用关系,重点阐述了沃尔巴克氏体与丝虫致病性和清除沃尔巴克氏体药物在人和动物丝虫病治疗中的作用,为人和动物丝虫病的防治研究提供参考。     

犬恶丝虫幼虫性皮肤病的诊治

本文描述一只3月龄沙皮犬患犬恶丝虫幼虫性皮肤病的诊断与治疗过程。治疗用依维菌素0.03mL／kg皮下注射1次，隔7天重复1次，以后3个月跟踪观察，未见复发。     

犬心丝虫致死病例一例

5～10月份是犬心丝虫病的高发季节。犬心丝虫病(carine dirofilariasis)又名犬恶丝虫病，是由犬心丝虫寄生于犬的右心室及肺动脉中引起循环障碍、呼吸困难及贫血等症状的一种丝虫病。除犬外，猫和其它野生肉食动物也可作为其终宿主。偶有寄生于马、海狸、猩猩和人。本病在日本的发病率很高。     

犬恶丝虫抗体dot-ELISA检测方法的建立

为快速诊断犬恶丝虫病,建立了犬恶丝虫抗体dot—ELISA检测方法,并与阻断试验、交叉反应试验、重复性试验、琼脂扩散试验和平行对照试验进行了比较。结果显示,制备的犬恶丝虫虫体粗制抗原与犬钩虫、犬弓首蛔虫和犬蠕形螨感染犬的阳性血清无交叉反应,检出犬恶丝虫阳性血清的最高抗体效价为1：1024,重复性好;建立的犬恶丝虫病dot—ELISA诊断方法的灵敏度比琼脂扩散试验和美国IDEXX实验室制备的犬恶丝虫病诊断试剂盒分别高2048倍和48倍。     

桑天牛卵啮小蜂孤雌生殖及体内沃尔巴克氏体(Wolbachia)的检测

桑天牛卵啮小蜂(Aprostocetus prolixus)有孤雌生殖现象,其后代几乎为雄性。沃尔巴克氏体(Wolbachia)是广泛分布于节肢动物体内的一类共生菌,参与多种调控寄主的生殖活动机制。通过对外膜蛋白基因(wsp)、细菌细胞分裂蛋白基因(ftsZ)和核糖体16S rDNA基因的特异性扩增,在米蛾(Corcyra cephalonica Stainton)的DNA中分别扩增出约600、1000和900 bp的片断,验证了米蛾体内已感染了Wolbachia;而在桑天牛卵啮小蜂的DNA中未扩增出任何片断,表明Wolbachia在桑天牛卵啮小蜂体内未被感染或感染率极低。     

昆虫共生菌沃尔巴克氏体(Wolbachia)的wsp基因密码子使用模式分析

沃尔巴克氏体(Wolbachia)主要感染昆虫等节肢动物,是一类革兰阴性共生细菌。为探讨昆虫纲共生菌Wolbachia的系统发育,研究Wolbachia基因组中进化速度较快的外膜蛋白基因wsp的密码子使用模式及影响因素,统计分析该基因的GC含量(GCall、GC1、GC2、GC3、GC3s)、有效密码子数(ENc)、相对同义密码子使用度(RSCU)等指标。结果显示:昆虫纲共生菌Wolbachia的wsp基因密码子偏好性均不强,昆虫纲不同目间共生菌Wolbachia的wsp基因碱基组成及ENc值差异较小;多数基因数据点沿标准曲线或在其附近分布,突变对碱基组成的影响较弱;18种由多个密码子编码的氨基酸中有11种氨基酸的偏好性密码子在昆虫纲7个目及对照蛛形纲1个目间均相同,有2种氨基酸的偏好性密码子在7个目间相同,这些密码子均以A/U结尾;供分析的140个wsp基因中仅编码6个半胱氨酸(Cys);对应分析中第1、第2向量轴贡献率均不高,分别为13.53%和12.49%,均与碱基组成(GC1、GC2、GC3)显著相关。综合各项分析认为,Wolbachia的wsp基因密码子偏好性不具有宿主分类特异性,密码子使用模式主要受碱基组成影响,而碱基组成主要受选择影响。     

犬恶丝虫微丝蚴超微结构的扫描电镜观察

通过扫描电镜和透射电镜可获得犬恶丝虫微丝蚴(Mf)超微结构的影像。横向环形条纹覆盖整个虫体的表面。观察头盘上的2个小孔和仅邻头盘第一条纹腹侧的口状腔。从上腭可投射出单个三角钩。观察前端第80环和后端第90环处的排泄孔。两处气孔位于虫体的腹侧。本研究首先证明大量核柱细胞分布于体腔。这些细胞呈球形,直径为1μm。每个细胞都包括球形核且彼此通过放射状微管连接。许多扁平肌细胞位于虫体皮下组织内部。尾部仅包括单个纵肌细胞。     

犬恶丝虫硫氧还蛋白过氧化物酶真核表达质粒的构建及初步免疫试验

为评价犬恶丝虫硫氧还蛋白过氧化物酶(TPx)真核表达质粒的免疫原性,本实验利用RT-PCR方法扩增TPx基因,将其克隆于真核表达载体pVAX1中构建重组质粒pVAX1-TPx,并对其进行体外表达鉴定及通过特异性抗体水平及相关的免疫因子的检测,评价其在体内诱导的免疫反应.实验结果表明,将pVAX1-TPx转染于Cos7细胞中能够正确表达TPx,其分子量约为28 ku,并被阳性血清所识别.将pVAX1-TPx免疫BALB/c小鼠并采用ELISA检测结果显示,重组质粒免疫组的外周血中抗体、Th2细胞分泌的IL4及IL13细胞因子水平均显著高于空质粒及空白对照组(p＜0.05);但Th1分泌的IFN-γ及IL2水平差异不显著.此外,淋巴细胞增殖试验结果也表明,pVAX1-TPx免疫组显著高于其他两个对照组(p＜0.05).实验数据表明pVAX1-TPx免疫可以有效诱导特异性的体液和细胞免疫.     

犬心丝虫病及其防治

犬心丝虫病是由恶丝虫属的犬恶丝虫寄生于犬的右心室和肺动脉所引起的临床或亚临床疾病。猫、狐、狼等动物亦能感染,人偶被感染。本病在我国分布很广,广东犬的感染率可达50%。1病原成虫细长,呈白色,食道长。雄虫长12~16厘米,尾端螺旋状蜷曲,有肛前乳突5对,肛后乳突6对,交合刺2根,不等长,左侧长,末端尖;右侧短,相当于左侧的1/2,末端钝圆。雌虫长25~30厘米,尾部     

东莞市犬心丝虫抗原、莱姆病抗体和犬埃里希抗体的调查报告

为了解心丝虫、莱姆病、犬埃里希氏体病在东莞市犬中的流行状况,对东莞市210份犬血清进行了心丝虫抗原、莱姆病抗体、犬埃里希抗体检测试验,试验结果:在检测的210份犬血清中,心丝虫抗原阳性0份、莱姆病抗体阳性0份,犬埃里希抗体阳性12份,犬埃里希抗体阳性率为5.71%。     

Comparison of serologic tests for detection of antigen in canine heartworm infections

In 30 random-source dogs, we determined sensitivity and specificity of 5 serologic tests for detection of canine heartworm antigens. Seventeen of the dogs were infected naturally with adult Dirofilaria immitis, and 4 of the infected dogs were amicrofilaremic. The ability of the serologic tests to predict whether a dog was infected or uninfected (overall test accuracy) ranged from 73 to 97%. Sensitivity was not affected by circulating D immitis microfilariae, but was markedly influenced by the number of adult D immitis present. False-positive reactions were rare and were not associated with intestinal parasites or Dipetalonema reconditum microfilariae. Modifications of some of the test procedures were necessary to maximize test accuracy and reproducibility. These modifications and other technical details might limit the usefulness of some of the tests in a veterinary practice.     

Comparison of serological tests for the detection of natural heartworm infection in cats

Serological tests were performed on 380 cats with necropsy-confirmed heartworm disease to compare the performance of currently available commercial laboratory and point-of-care heart-worm serological tests in a heartworm-endemic area. Overall, antigen tests detected 79.3% to 86.2% of heartworm infections and were highly specific. Most cats with false-negative antigen tests had a single male worm. Antibody tests detected 62.1% to 72.4% of heartworm infections and had a wider range of false-positive results (1.4% to 19.1%) than antigen tests (0.3% to 2.0%). Serological tests for feline heartworm infection varied in diagnostic performance. Combining results from antigen and antibody tests achieved greater sensitivity than using either test alone.     

Evaluation of a new in-clinic method for the detection of canine heartworm antigen

Canine heartworm is endemic in many parts of the world, and veterinarians rely on rapid in-clinic antigen tests to screen for this infection. Recently, an in-clinic, instrument-based rotor employing a colloidal gold agglutination immunoassay was launched in the marketplace (VetScan VS2(?) Canine Heartworm (HW) Antigen Test Kit; Abaxis, Inc.). Because of the widespread use of heartworm prevention and possible false negative test results in dogs with low heartworm burdens, the performance of the VetScan VS2(?) HW test and a commercially available in-clinic, membrane-based ELISA test (SNAP(?) Heartworm RT Test; IDEXX Laboratories) was compared using samples from dogs with low heartworm burdens and/or low levels of circulating antigen. Ninety serum samples were evaluated using the two methods. Testing was performed according to the manufacturer's product insert by personnel blinded to sample status. The samples were derived from two populations: dogs with necropsy-confirmed heartworm status (40 with 1-4 female worms, 30 with no worms), and field dogs (20) confirmed positive for antigen by microtiter plate ELISA (PetChek(?) Heartworm PF Antigen Test; IDEXX Laboratories). All 40 dogs with heartworms on necropsy were also confirmed to have circulating antigen by the PetChek HW ELISA. In necropsy-negative dogs (n=30), neither the VetScan VS2 HW nor SNAP HW tests detected heartworm antigen. Of the samples testing positive for antigen by PetChek HW (n=60), the VetScan VS2 HW and SNAP HW tests detected antigen in 15 and 56 samples, respectively. Percent agreement (plus 95% confidence interval) for each test relative to the PetChek HW qualitative result was 50% (40-60%) for VetScan VS2 HW and 96% (89-98%) for SNAP HW. Relative to the presence or absence of female worms at necropsy, agreement was 61% (50-72%) for VetScan VS2 HW and 99% (92-99.6%) for SNAP HW tests. It is clinically important that dogs with low heartworm burdens and/or low levels of circulating heartworm antigen be correctly identified by veterinarians in order to ensure prompt treatment, and the VetScan(?) VS2 HW test does not appear to be as accurate as the SNAP HW or PetChek HW tests when performed on this subset of patients.     

Comparison of heartworm antigen test kit performance in dogs having low heartworm burdens

Sensitivity and specificity of four in-clinic heartworm antigen test kits, AbboScreen (Abbott Laboratories), Snap PF (IDEXX Laboratories), Solo Step (HESKA Corporation), Witness (Synbiotics Corporation) and two heartworm antigen microwell plate assays, DiroCHEK (Synbiotics) and PetChek PF (IDEXX) were compared in a blinded study using serum or plasma drawn from 237 random source dogs, including 140 with necropsy-confirmed, low worm burden infections (minimum 1 worm, maximum 10, mean 2.3, median 3) and 97 confirmed heartworm-free at necropsy. In general, microwell format tests were more sensitive than membrane format tests and tests using ELISA technology were more sensitive than tests using lateral flow immunochromatographic technology. Percent sensitivity and specificity, respectively, were PetChek PF 76 and 97, DiroCHEK 71 and 94, SNAP PF 67 and 98, Solo Step 60 and 98, and AbboScreen 52 and 96. The Witness test protocol was changed by the manufacturer midway through the study, and the newer version of this test kit arrived containing a package insert alerting the user to a change in procedure, which purportedly resulted in improved sensitivity. PetChek was significantly more sensitive than all other tests except DiroCHEK and the new version of Witness. DiroCHEK was significantly more sensitive than all tests except PetCheck, SNAP and the new version of Witness. Snap was more sensitive than AbboScreen and the old version of Witness. Differences in specificity were not significant (P>0.05).