南宁市市售猫华支睾吸虫感染调查

目的　了解南宁市猫华支睾吸虫感染情况，为华支睾吸虫保虫宿主防控提供依据。方法　从南宁市猫屠宰点购买猫肝，解剖胆囊和肝脏，查找华支睾吸虫并计数；对收集到的不同形态和大小的华支睾吸虫进行形态学观察和ITS2基因扩增和测序，通过BLAST比对分析确定虫体种类。结果　从南宁市2个猫屠宰摊面共收集了105个猫肝脏，其中有68个发现有华支睾吸虫感染，感染率为64.76%。单个肝脏虫体数量最多达980条，虫体数量中位数为72条/个肝脏。从检出的虫体中分离到3种不同大小和形态的吸虫，经形态学和ITS2基因扩增、测序、比对分析鉴定，均为华支睾吸虫。结论 　南宁市市售猫华支睾吸虫感染率较高，应该加强相关防控措施。     

家猫感染东方次睾吸虫形态学特征及分子鉴定

目的 了解家猫感染东方次睾吸虫的形态学及基因特征，为当地开展东方次睾吸虫病防控工作提供理论依据。方法 采集浦城县感染有东方次睾吸虫囊蚴的麦穗鱼，剪碎囊蚴寄生处的鱼尾部肌肉，喂食2个月龄家猫。45 d后连续多天检测家猫粪便中的虫卵，直至发现虫卵后进一步深入检查。观察肝脏及胆囊变化和虫体寄生部位，捡出虫体置于生理盐水中，对虫体的形态学特征进行观察与鉴定描述。提取虫体基因组DNA,PCR扩增ITS基因序列，测序后在GenBank上进行BLAST比对，采用邻接法构建系统进化树。结果 从胆囊和肝脏分离获得13条东方次睾吸虫。东方次睾吸虫为雌雄同体，虫体大小约为(1.95～2.86)mm×(0.61～0.93)mm;虫卵呈长椭圆形，似葵花籽状，虫卵大小约(28～32)μm×(16～18)μm。PCR结果显示，ITS基因序列扩增产物260 bp。BLAST比对结果显示，与国内已报道家鸭及黑天鹅感染的东方次睾吸虫(GenBank登入号分别为MT231323.1、MK482055.1、MK482052.1)的序列一致性均为99.62%。基于ITS基因的系统进化树显示，本研究分离获得的吸虫ITS序列属于次...     

嫩江流域齐齐哈尔段犬、猫华支睾吸虫感染情况调查

目的了解嫩江流域齐齐哈尔段保虫宿主犬和猫的华支睾吸虫感染情况,为华支睾吸虫病的防治提供参考依据。方法从嫩江流域齐齐哈尔段周边村庄及河岸边采集犬和猫的粪便,在周边县市的动物屠宰场收集犬的肝脏及胆囊:采用改良加藤法检查犬和猫粪便中华支睾吸虫虫卵的感染情况;采用解剖法检查犬的肝脏及胆囊内华支睾吸虫成虫的感染情况。结果嫩江流域齐齐哈尔段华支睾吸虫保虫宿主粪便虫卵的总感染率为8.00%(4/50),其中犬粪虫卵感染率为8.57%(3/35),猫粪虫卵感染率为6.67%(1/15);解剖法发现华支睾吸虫保虫宿主犬的成虫感染率为5.71%(2/35)。结论嫩江流域齐齐哈尔段为肝吸虫病流行较为严重地区,建议加大查治和健康教育等多种防控措施力度,减少华支睾吸虫病对嫩江流域人民的健康危害。     

南昌市鸭源吸虫形态与分子鉴定

目的　对分离自南昌市蛋鸭体内的吸虫进行形态学和分子生物学鉴定。方法　从南昌市市售蛋鸭肝胆管、胆囊和大肠中分离吸虫。经形态学鉴定后,提取所有吸虫样本总DNA,PCR扩增内转录间隔区（ITS）和细胞色素氧化酶亚基Ⅰ（Cox1）基因并测序,序列经Blast比对后,分别对吸虫样本ITS和Cox1基因进行同源性和系统发育分析。结果　2条分离自蛋鸭大肠的吸虫样本与卷棘口吸虫和宫川棘口吸虫形态特征相似,8条分离自蛋鸭肝胆管和胆囊的吸虫样本形态与对体吸虫相似。2条分离自蛋鸭大肠的吸虫样本ITS和Cox1基因序列与宫川棘口吸虫同源性分别为99.3%和98.9%～99.4%,基于ITS和Cox1基因序列的系统进化树显示吸虫样本与宫川棘口吸虫亲缘关系最近。8条分离自蛋鸭肝胆管和胆囊吸虫样本ITS基因序列与苏氏肝居吸虫和华支睾吸虫同源性分别为95.1%～95.5%,Cox1基因序列与麝后睾吸虫和苏氏肝居吸虫同源性分别为86.3%～86.4%和85.5%～85.7%。基于ITS和Cox1基因序列的系统进化树显示,吸虫样本ITS基因序列与华支睾吸虫亲缘关系最近,Cox1基因序列与东方次睾吸虫和麝后睾吸虫亲缘关系最近。 结论　自南昌市蛋鸭大肠分离的吸虫可能为宫川棘口吸虫,分离自蛋鸭肝胆管和胆囊的吸虫可能为一种未定种属的后睾科吸虫。     

华支睾吸虫病患者粪便虫卵核糖体DNA内转录间隔区序列分析

目的 通过PCR方法扩增华支睾吸虫病患者粪便虫卵核糖体DNA （ribosomal DNA,rDNA）内转录间隔区（internal transcribed spacer,ITS）序列进行分析,探讨其用于华支睾吸虫感染检测的价值. 方法 提取2例华支睾吸虫患者粪便中的虫卵DNA作模板,以rDNA ITS基因片段为目的片段进行PCR扩增,对扩增片段进行测序分析. 结果 从2例患者粪便虫卵样本中均扩增出1 123 bp的条带,经序列比对确定患者粪便中的虫卵为华支睾吸虫卵.测序分析发现本研究的两个样本与中国黑龙江分离株（KF740425）相似性为100％. 结论 本研究从2例华支睾吸虫病患者粪便虫卵中成功扩增出rDNA ITS基因,为从分子生物学角度检测华支睾吸虫病提供了资料.     

广州珠三角地区家猫自然感染华支睾吸虫的调查及豚鼠模型的构建北大核心CSCD

目的了解广州珠三角地区家猫自然感染华支睾吸虫状况,观察豚鼠感染华支睾吸虫后肝组织病理变化。方法解剖当地家猫,取肝脏,检查华支睾吸虫感染情况,阳性肝脏收集成虫,并取肝组织制作切片;采集阳性鱼,分离华支睾吸虫囊蚴,经口感染豚鼠,60个囊蚴/只,60d后解剖豚鼠,检查肝脏,收集成虫,取病变肝组织制作切片,作病理检查。实验设未感染对照组。结果当地猫华支睾吸虫自然感染率为41.47%(214/516);感染豚鼠华支睾吸虫成虫回收率50.83%(305/600)。与对照组比较,实验组豚鼠肝脏肿大,边缘呈球状隆起,部分肝叶表面可见水泡状凸起病变,水泡内液清亮;病变组织横切面可见胆管管壁增厚,管腔内有华支睾吸虫成虫寄生;镜下可见华支睾吸虫虫卵切面,周围大量炎症细胞浸润,胆管管壁增厚,管周纤维组织增生伴胆管扩张,肝小叶结构破坏。结论广州珠三角地区家猫华支睾吸虫感染率较高。豚鼠是华支睾吸虫合适的终末宿主,且肝脏病变明显。     

广州珠三角地区家猫自然感染华支睾吸虫的调查及豚鼠模型的构建

目的了解广州珠三角地区家猫自然感染华支睾吸虫状况,观察豚鼠感染华支睾吸虫后肝组织病理变化。方法解剖当地家猫,取肝脏,检查华支睾吸虫感染情况,阳性肝脏收集成虫,并取肝组织制作切片;采集阳性鱼,分离华支睾吸虫囊蚴,经口感染豚鼠,60个囊蚴／只,60d后解剖豚鼠,检查肝脏,收集成虫,取病变肝组织制作切片,作病理检查。实验设未感染对照组。结果当地猫华支睾吸虫自然感染率为41．47％（214／516）;感染豚鼠华支睾吸虫成虫网收率50．83％（305／600）。与对照组比较,实验组豚鼠肝脏肿大,边缘呈球状隆起,部分肝叶表面可见水泡状凸起病变,水泡内液清亮;病变组织横切面可见胆管管壁增厚,管腔内有华支睾吸虫成虫寄生;镜下可见华支睾吸虫虫卯切面,周围大量炎症细胞浸润,胆管管壁增厚,管周纤维组织增生伴胆管扩张,肝小叶结构破坏。结论广州珠三角地区家猫华支睾吸虫感染率较高。豚鼠是华支睾吸虫合适的终末宿主,且肝脏病变明显。     

广西藤县人群华支睾吸虫感染调查及其分子鉴定

目的了解广西藤县人群华支睾吸虫的感染状况及其分子鉴定,为当地华支睾吸虫病的防控提供理论依据。方法2016年11月,在广西藤县采用分层整群随机抽样抽取天平镇和金鸡镇各1个自然村作为调查点。收集3岁以上常驻居民的粪便,用改良加藤厚涂片法(一粪三检)检测华支睾吸虫虫卵并计数。通过问卷调查收集被调查者社会人口学等相关信息。从镜检阳性样本中随机选取部分新鲜粪样提取DNA, PCR扩增华支睾吸虫内转录间隔区2 (ITS2),测序后在NCBI核酸数据库作比对分析。结果共调查400人,检出华支睾吸虫虫卵阳性208人,感染检出率为52.0%(208/400),感染者以轻度感染为主(69.2%, 144/208)。天平镇华支睾吸虫感染检出率为66.5%(133/200),高于金鸡镇的37.5%(75/200)(χ2=33.69, P 0.01)。男性感染检出率为58.7%(131/223),高于女性的43.5%(77/177)(χ2=9.18, P 0.01); 30～39岁组感染检出率最高,为63.8%(60/94);高中及以上学历人群感染检出率最高,为68.0%(17/25)。不同年龄组、教育程度人群感染检出率差异有统计学意义(χ2=37.1, 23.02; P 0.01)。PCR扩增出156 bp的华支睾吸虫特异性条带,其序列与越南岘港分离株ITS2 (GenBank登录号:MF319655)的一致性为100%。结论广西藤县人群华支睾吸虫感染检出率为52.0%,属于超重流行区;男性、青壮年是华支睾吸虫高感染人群。广西藤县的华支睾吸虫ITS2序列与越南岘港分离株的基因型一致。     

家猫检获大量华支睾吸虫1例

华支睾吸虫成虫常寄生于人体肝胆管内,也寄生于狗、猫等动物的肝、胆囊及胆管,可引起华支睾吸虫病——一 种重要的人兽共患寄生虫病。本研究通过解剖1例家猫,从其肝脏和胆囊中检获虫体共736条,经鉴定为华支睾吸虫。 这一发现提示芜湖地区有动物华支睾吸虫病流行,应重视宠物寄生虫感染的防治。     

华支睾吸虫cDNA文库的构建

目的　构建华支睾吸虫cDNA文库。方法　采集华支睾吸虫阳性鱼 ,分离囊蚴 ,感染实验动物兔 ,解剖兔收集华支睾吸虫成虫虫体。应用“一步法”提取华支睾吸虫总RNA ;经过mRNA纯化、cDNA合成 ,以PcDNA3(Amp +)质粒为载体构建文库。挑取 2 0个克隆进行扩增 ,选择 9个克隆进行DNA序列测定。序列结果与GenBank中相关基因进行比对。结果　获得含有 9 7× 10 5个重组子库容量的华支睾吸虫cDNA文库。其中PC6号克隆基因序列与华支睾吸虫半胱氨酸蛋白酶序列具有 93%的同源性。结论　已构建成华支睾吸虫cDNA文库。     

Discovery of Opisthorchis viverrini metacercariae in freshwater fish in southern Cambodia

Small liver flukes, Clonorchis sinensis and Opisthorchis viverini, are fish-borne trematodes (FBTs) causing significant public health problems in Asia. While C. sinensis is distributing mainly in far east Asia, O. viverini is distributing in Indochina peninsula. Recently, however, the geographical distributions of those small liver flukes were proven to be far wider than expected. Nevertheless, little is known about the epidemiology of small liver flukes in Cambodia. The present study is, therefore, aimed at clarifying the status of small liver fluke infections in various species of freshwater fish in southern Cambodia. A total of 1479 freshwater fish, 1316 (89%) comprised of 20 different species of the cyprinoid family and 163 (11%) belonging to 8 families of non-cyprinoids, were collected during May 2007 and February 2008, and the presence of small liver fluke metacercariae was examined by the compression method. Small liver fluke metacercariae were found in 10 species of cyprinoids with the infection rate ranging 2.1-66.7% and the mean intensity of infection of 1.0-15.0 (range: 1-65). For the speciation, adult worms were obtained by experimental infection in hamsters. The small liver fluke found in this study were identified as Opisthorchis viverrini by the morphological features of adult worms, and this identification was confirmed by partial COI sequencing of the metacercariae.     

Trematode infections: liver and lung flukes

Food-borne trematodiases are an emerging public health problem in Southeast Asia and Latin America and of growing importance for travel clinics in Europe and North America. The disease is caused by chronic infections with liver, lung, and intestinal flukes. This article focuses on the most important liver and lung flukes that parasitize man, namely Clonorchis sinensis, Fasciola gigantica, Fasciola hepatica, Opisthorchis felineus, Opisthorchis viverrini, and Paragonimus spp. The article describes the epidemiology of major liver and lung fluke infections, including current distribution, burden, life cycle, clinical signs and symptoms, diagnostic approaches, and current tools for prevention, treatment, and control.     

Geographical variation of the liver fluke,Clonorchis sinensis,from Korea and China based on the karyotypes,zymodeme and DNA sequences

Genetic characterization was carried out in order to reveal the geographical variations of the oriental liver fluke, Clonorchis sinensis (Trematoda: Opisthorchiidae), collected in Korea and China. The chromosome number was 2n=56 in both Korean (Kimhae) and Chinese (Liaoning) flukes, and chromosomes were divided into two groups based on their sizes; consisting of 8 pairs of large and 20 pairs of small chromosomes. However, the karyotypes showed some differences between Korean and Chinese flukes. Isozyme analysis showed that two loci from each enzyme of aconitase and esterase (alpha-Na and beta-Na); only one locus each from six enzymes, glucose-6-phosphate dehydrogenase (G6PD), alpha-glycerophosphate dehydrogenase (GPD), 3-hydroxybutyrate dehydrogenase (HBDH), malate dehydrogenase (MDH), phosphoglucose isomerase (PGl) and phosphoglucomutase (PGM). Most of loci in two populations of C. sinensis showed homozygous monomorphic banding patterns and one of them, GPD was specific as genetic markers between two different populations. Two populations were very closely clustered within the range of genetic identity value of 0.998-1.0. We also compared patterns of intraspecific polymorphism of two markers with contrasted modes of evolution, nuclear ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) of the liver fluke from Kimhae, Guangxi and Liaoning. They showed a high homology. In conclusion, three populations of C. sinensis from Korea and China showed high homogeneity in the nucleotide sequences of the 18S rDNA, ITS2 and mtCOI gene.     

虫卵ITS-2序列分析鉴别华支睾吸虫和扇棘单睾吸虫的研究

目的建立以分子生物学技术为基础的华支睾吸虫与扇棘单睾吸虫的ITS-2序列分析鉴别方法。方法在华支睾吸虫流行区采集华支睾吸虫和扇棘单睾吸虫成虫,从成虫虫体分离并收集虫卵。以蛔虫、鞭虫、钩虫、蛲虫做对照虫种,按不同虫种和虫卵数分成5个实验组,各组分别提取DNA,并扩增ITS-2序列,对扩增产物进行测序并作同源性分析以确保为目标片段,根据PCR扩增产物电泳获得的条带判定虫种。结果以华支睾吸虫和扇棘单睾吸虫虫卵DNA为模板的PCR产物电泳图谱分别在400bp、500bp左右各显示一条明显条带;在以两种虫卵DNA混合液为模板的PCR产物中,电泳图谱在400bp左右和500bp左右各显示明显条带。将测序得到的两种核苷酸序列进行Blast比对后,显示与GenBank中相应的华支睾吸虫和扇棘单睾吸虫序列高度同源。混入四种肠道线虫虫卵DNA的PCR产物中,除目的条带外,无其他条带。结论该方法可以对华支睾吸虫及扇棘单睾吸虫虫卵进行鉴别,且结果不受常见四种肠道线虫虫卵干扰,敏感性和特异性较高,可作为两虫种的鉴别检测方法。     

Parasitic infections and hepatic neoplasia

Parasitic infections are highly prevalent in the general population. Detecting a relationship between a parasitic infection and cancer is not an easy task; it requires epidemiological, microbiological and molecular biology techniques. In this article the important mechanisms by which parasitic infections may induce carcinogenesis are reviewed. The association between Schistosoma japonicum, Schistosoma mansoni and hepatocellular carcinoma is covered. The cause-and-effect relationship between the liver flukes Opisthorchis viverrini, Opisthorchis felineus, Clonorchis sinensis, and cholangiocarcinoma is also reviewed. The evidence supporting the role of schistosomes and liver flukes in carcinogenesis is discussed. Finally the importance of the primary prevention of these parasitic infections with oncogenic potentials is emphasized.     

广东省三水市华支睾吸虫病的防治研究

１９８６－１９８８年和１９９０－１９９２年分别在本市董永区和官元区，以反复查治病原方法防治华支睾吸虫病，并对中间宿主及保虫宿主进行调查。经两次查治后，人群华支睾感染率董永区从４３．６％降为１３．２％，官元区从７８．５％降为３６．３％。当地第一中间宿主是长角涵螺，第二中间宿主是鲸鱼和鳊鱼。在猫粪便中发现华支睾吸虫虫卵，在狗和猪胆管内均发现华支睾吸虫卵及成虫。