Oxfendazole as successful treatment of Taenia hydatigena metacestodes in naturally infected pigs

The efficacy of oxfendazole(OFZ) on Taenia hydatigena metacestodes, also called Cysticercus tenuicollis(C. tenuicollis), was studied in 648 raising pigs. This study was performed in Tumbes Department in Peru, an endemic area for cysticercosis. Pigs were randomized in two groups; untreated group(n = 142) did not receive any treatment and treated group(n = 506) received OFZ treatment at a single dose of 30 mg/kg body weight.Six months after treatment, the pigs were necropsied. The prevalence of infection by C. tenuicollis among the pigs was 27.5%(39/142) and 2.0%(10/506) in untreated and treated groups, respectively. Untreated group was infested only with viable cysts, whereas treated group had no viable cysts. All the cysts found in treated group presented degeneration, with a thick membrane, and they contained milky fluid and fibrous tissue. A single dose of OFZ was effective against C. tenuicollis, thus providing an alternative drug for controlling this parasite in pigs.

Genetic variation of the 8-kDa glycoprotein family from Echinococcus granulosus, Taenia multiceps and Taenia hydatigena

Background Echinococcosis, coenurosis and cysticercosis are debilitating diseases which prevail in China. Immunological diagnosis of metacestodosis is important in disease control. The 8-kDa glycoproteins from taeniid cestodes have successfully been used for diagnosis of human cysticercosis in immunological assays. The aim of the present study was to investigate genetic variations and phylogenetic relationships of the 8-kDa proteins for evaluating the possibility of utilizing these proteins as diagnostic antigens for other metacestode infections. Methods The genes and complementary DNAs （cDNAs） encoding the 8-kDa proteins from Echinococcus （E.） granulosus, Taenia （T.） multiceps and T. hydatigena were amplified using PCR method. Their amplicons were cloned into the vector pMD18 and the positive clones were sequenced. Sequence data were analyzed with the SeqMan program, and sequence homology searches were performed using the BLAST program. Alignments were conducted using the ClustalX program, and the phylogenetic analyses were performed with the Protein Sequences Program and the Puzzle Program using the Neighbor-joining method. Results Fifteen, 18 and 22 different genomic DNA sequences were identified as members of the 8-kDa protein gene family from E. granulosus, T. multiceps and T. hydatigena, respectively. Eight, four and six different cDNA clones respectively from E. granulosus, T. multiceps and T. hydatigena were characterized. Analysis of these sequences revealed 54 unique 8-kDa protein sequences. Phylogenetic trees demonstrated that the taeniid 8-kDa proteins are clustered into eight clades at least： Ts18, Ts14, TsRS1, TsRS2, T8kDa-1, T8kDa-2, T8kDa-3 and T8kDa-4. Conclusion We found that the gene family encoding for the taeniid 8-kDa antigens is comprised of many members with high diversity, which will provide molecular evidence for cross-reaction or specific reaction among metacestode infections and may contribute to the development of promising immunological methods for diagnosis of metacestodosis.

犬泡状带绦虫线粒体cox1和nad1基因的扩增与种系发育分析

以采自广州和佛山的犬泡状带绦虫为研究对象,利用PCR技术扩增其线粒体基因组的细胞色素c氧化酶第亚基（cox1）和烟酰胺腺嘌呤二核苷酸还原酶第一亚基（nad1）序列,扩增产物经纯化、克隆和测序。结果扩增出泡状带绦虫线粒体cox1基因序列长度为444 bp,nad1基因序列长度为530 bp。将实验获得序列与GenBank收录的相关序列进行比对分析,发现采自不同国家和地区的泡状带绦虫cox1基因差异度较小（02.3%）,而nad1基因的差异度较大（0.214.2%）,对进一步研究泡状带绦虫的群体遗传结构奠定了基础。

泡状带绦虫线粒体部分基因克隆及序列比对分析

采集甘肃兰州市区宠物犬体内两株泡状带绦虫,运用分子生物学方法对其线粒体部分基因进行克隆,并使用基因分析软件Larsergene V7.1进行序列分析,结果发现这两株虫体同源性达到99.6%,与国内的泡状带绦虫广东虫株和甘肃虫株同源性分别达到98.7%和98.4%,鉴定为泡状带绦虫;与其他带科绦虫同源性均高于85%,低于88%。其中,与国内牛带绦虫同源性最高,为88%;与国内巨颈带绦虫同源性最低,只有85.1%。推断不同带科绦虫的cox1基因种间差异明显,在带科绦虫的分类上意义较大。

湖南省泡状带绦虫线粒体nad1基因的序列测定及分析

以从我国湖南长沙和湘西犬小肠中采集的2条泡状带绦虫作为研究对象,用引物JB11及JB12扩增泡状带绦虫的pnad1片段,应用ClustalX 1.81程序对序列进行比对,同时利用DNAstar5.0中的Megalign程序进行同源性分析。结果显示来自湖南长沙和湘西的2条泡状带绦虫的pnad1序列均为391bp。研究结果为泡状带绦虫进一步的分类、鉴定和遗传变异研究奠定了基础。

泡状带绦虫线粒体ND1基因的克隆及序列比对分析

针对带绦虫线粒体DNA的ND1基因设计引物扩增目的基因,使用基因分析软件Larsergene V7.1与GenBank已知带绦虫基因进行比对分析。实验成功扩增泡状带绦虫甘肃地区虫株X2线粒体基因组的ND1基因。分析结果显示,泡状带绦虫ND1基因种间不同虫株的基因同源性均在98.8%~99.5%之间,ND1基因推导的氨基酸序列与其他泡状带绦虫的蛋白同源性均在98.5%~99.2%之间,其中与四川地区的虫株同源性均达到了99.2%。推断泡状带绦虫线粒体DNA的ND1基因相对保守,与Taenia sp.AL-2012、Taenia regis和猪带绦虫差异较明显,最高只有91.3%,可以作为泡状带绦虫的初步鉴别基因。

犬泡状带绦虫潜在前期研究与PCR鉴定

为查明猪细颈囊尾蚴在犬体内发育为成虫后节片的最早排出时间(潜在前期),将新鲜的猪细颈囊尾蚴经口感染6只无绦虫犬后,采用反复水洗沉淀法检测感染犬粪中排出的第1个绦虫节片,并用特异PCR方法鉴定其种属。结果显示,6只感染犬的潜在前期分别为60、63、65、69、72、73 d。同时,特异性PCR结果显示其序列与Genbank收录的泡状带绦虫序列的相似性为99%,确认犬排出的节片为泡状带绦虫。本研究为建立泡状带绦虫在宿主体内的发育模型提供了一定的基础资料。

催产素和加压素在犬泡状绦虫的免疫组化定位和分布

应用免疫组化ＡＢＣ法研究了催产素和加压素在犬泡状绦虫的定位和分布。在犬泡状绦虫中枢神经系统的脑神经节、２条主要纵神经索和环神经内观察到大量阳性纤维和少量阳性细胞。周围神经系统可见阳性神经纤维在皮层和实质形成神经网。从纵神经索和环神经发出的阳性纤维向生殖器官和皮层的肌肉分布。顶突的小钩之间的肌肉存在阳性反应物，小钩周围有较明显的阳性反应环。生殖器官呈现着色深浅不同的阳性反应，以卵囊—梅氏腺复合体最为明显。结果表明，催产素和加压素的免疫反应物具有相同的分布，它们可能参与虫体的神经内分泌调节。

Molecular characterization of Cysticercus tenuicollis of slaughtered livestock in Upper Egypt governorates

Objective: To present the molecular characterization of Cysticercus tenuicollis(C. tenuicollis) of Taenia hydatigena(T. hydatigena) from livestock isolates in Egypt, and to introduce a detailed image of C. tenuicollis infection in ruminant animals in Upper Egypt.Methods: The prevalence rates of C. tenuicollis infections among the slaughtered animals from different organs were determined using the amplification of sequencing of the MT-CO1 gene.Results: In the present study the infection rates of C. tenuicollis were found to be 16%and 19% in sheep and goat samples respectively. Firstly we report one larval stage of T. hydatigena detected in the camel liver in Egypt. C. tenuicollis infection manifested a higher prevalence in females than in males. Those above two years of age manifested a higher infection rate than younger animals. The preferred site for the infection was the omentum: a 70% preference in sheep and a 68% preference in goats. The molecular characterization using the MT-CO1 gene of isolates from sheep, goats and camels corresponded to T. hydatigena. For this study, molecular characterizations of T. hydatigena were done for the first time in Egypt. Molecular tools are of great assistance in characterizing the C. tenuicollis parasite especially when the morphological character cannot be detected, because the metacestodes are frequently confused with infection by the hydatid cyst, especially when these occur in the visceral organs. In the present study,C. tenuicollis manifested high identity in the goat and sheep samples, while differences were found more frequently in the camel samples(10 base pair).Conclusions: Clearly molecular diagnosis for C. tenuicollis infection significantly helps to differentiate it from such other metacestodes as hydatidosis, which manifests a completely different pathogenicity and requires different control programs.

Incidental findings of Cysticercus tenuicollis metacestodes in five oryx species

Objective:To determine the prevalence of Cysticercus tenuicollis(C.tenuicollis) metacestodes in five oryx species kept in A1 Ain Zoo animal collection.Methods:This study was based on a retrospective analysis of post-mortem records covering a four year period(July 2010 to July 2014).Results:A total of 213 individual animals were recorded dead during the four year period(July 2010 to July 2014).Out of this,12(5.6%) were recorded with C.tenuicollis.More females(8) than males(4) were recorded to have C.tenuicollis,although this was not statistically significant(P= 0.3737).Conclusions:This study shows that,Arabian oryx,beisa oryx,fringe-eared oryx,gemsbok and scimitar-horned oryx are susceptible to C.tenuicollis.Based on the epidemiology and the life cycle of this parasite,it is possible that these captive animals ingested the parasite through contaminated feed which could have happened in the pasture land or stray dogs and wild canidae(e.g.fox) visited the zoo contaminating the oryx feed.Stray dogs and wild canidae should be prevented from visiting pasture land and a captive animal facility.

湖南长沙及湘西泡状带绦虫分离株线粒体cox1基因的克隆及序列分析

目的研究湖南省长沙及湘西泡状带绦虫分离株线粒体细胞色素c氧化酶第I亚基(cox1)基因部分序列(pcox1)的遗传变异情况,并用pcox1序列重构泡状带绦虫与其他带科绦虫的种群遗传关系。方法应用聚合酶链反应(PCR)扩增泡状带绦虫虫株的pcox1,应用ClustalX1.81程序对序列进行比对,然后用Phylip3.67程序MP法和Mage4.0程序NJ法绘制种系发育树,并用Puzzle5.2程序构建最大似然树;同时利用DNAstar5.0中的Megalign程序进行同源性分析,并与GenBankTM中已知泡状带绦虫相应基因序列进行比较分析。结果所获得的pcox1序列长度一致,均为343bp,与GenBankTM公布的带科绦虫序列进行比较分析表明,湖南长沙分离株1和湘西分离株5与已知泡状带绦虫相应基因的相似性分别为99.4%和99.7%,与其它带科绦虫的相似性均小于90.0%;湖南长沙1和湘西分离株5与已知泡状带绦虫相应基因遗传距离分别为0.006和0.003,与其他带科绦虫的遗传距离均大于0.100。种系发育树表明,2个分离株与已知泡状带绦虫位于同一分枝,Bootstrap值在97%以上。结论由于泡状带绦虫pcox1序列种内相对保守,种间差异较大,故可作为种间遗传变异研究的标记,研究结果为进一步研究泡状带绦虫的群体遗传结构奠定了基础。

水泡带绦虫海南株的分子鉴定及系统发育分析

目的以COI基因和ND1基因作为DNA条形码对水泡带绦虫海南株进行分子鉴定和系统发育分析。方法PCR扩增COI基因和ND1基因片段,GenBank数据库中进行BLAST多序列比对确定虫种。使用MEGA11软件构建基于邻接法的COI基因和ND1基因系统发育树,并计算基于K2P模型的种内遗传距离和种间遗传距离;分析核苷酸组成,计算碱基转换率、碱基颠换率及碱基转换与颠换比值。使用DNAsp软件分析COI基因和ND1基因的单倍型多样性。登录ABGD网站检测COI基因和ND1基因是否存在明显的DNA barcoding gap,以评估COI基因和ND1基因作为DNA条形码进行物种分子鉴定的有效性。利用DAMBE软件检测碱基替换饱和度,以明确COI基因和ND1基因构建系统发育树的准确性。结果COI基因的单倍型多样性为0.8276,含有81个可变位点,碱基转换率(si)为10%,碱基颠换率为(sv)13%,转换与颠换比值为0.77。ND1基因的的单倍型多样性为0.9381,含有94个可变位点,碱基转换率(si)为10%,碱基颠换率(sv)为11%,转换与颠换比值为0.91。COI基因和ND1基因均具有明显的DNA barcoding gap,可作为有效的DNA条形码进行带绦虫属内物种的分子鉴别。COI基因、ND1基因Iss<Iss.c,且P<0.05,两者均不存在碱基替换饱和,包含了足够的系统发育信息,适合进行系统发育分析。结论单基因作为DNA条形码能够快速、简便的鉴别物种,而多重DNA条形码能够提高分辨率和计算效率,是分析系统发育关系和探索生物多样性的有效识别工具。