云南省怒江州部分地区独龙牛毕氏肠微孢子虫感染情况及基因分型研究

为了解云南省怒江州独龙牛毕氏肠微孢子虫感染情况,从怒江州的鸠门当、古泉村、亚左洛村、茨开镇、独龙江乡分四个季度(春、夏、秋、冬)采集独龙牛粪便样本1 129份,采用分子生物学技术对其毕氏肠微孢子虫感染情况进行调查研究.结果发现,所调查地区独龙牛毕氏肠微孢子虫总感染率为1.68％(19/1 129);各地点之间感染率差异...     

江苏和云南部分地区山羊毕氏肠微孢子虫感染情况调查

为了调查江苏省和云南省部分地区山羊毕氏肠微孢子虫的感染情况和基因型,从江苏4个地区和云南9个地区羊场采集282份山羊新鲜粪样,提取基因组DNA,采用基于毕氏肠微孢子虫核糖体内转录间隔区(ITS)序列的特异套式PCR方法进行检测,对阳性扩增产物进行克隆、测序分析,以确定其基因型。结果显示,仅在云南昆明、江苏镇江和宿迁地区的山羊粪样中检出毕氏肠微孢子虫,各地区的感染率为0～22.7%。总感染率为2.8%(8/282),江苏感染率为6.3%(6/94),云南感染率为1.1%(2/188)。不同月龄羊间感染率差异不显著(P>0.05)。阳性扩增产物经克隆、测序分析,共获得2个基因型CHG1(n=3)和CHG3(n=5),进化树分析显示2个基因型均属Group 2亚群,无人畜共患性。     

秦岭地区几种珍稀动物毕氏肠微孢子虫基因分型

为了评估秦岭地区珍稀野生动物的毕氏肠微孢子虫(Enterocytozoon bieneusi)的人兽共患风险,本研究采用PCR技术和多位点序列分型技术对秦岭地区5种珍稀动物毕氏肠微孢子虫的感染情况及基因型/亚型进行了分析。在采集的22份新鲜粪便中检测到7个阳性分离株,其中斑羚源5个,长颈鹿源1个,麂子源1个。基于ITS位点基因分型发现所有分离株均为人兽共患基因型,包括6个已报道的基因型D和1个尚未报道的D-new。多位点序列分型技术发现秦岭地区珍稀野生动物的毕氏肠微孢子虫存在遗传多样性,7个分离株在MS1、MS3、MS4和MS7位点分别有3、1、2、2个亚型。     

云南省部分地区荷斯坦牛毕氏肠微孢子虫感染情况调查及分析

为了解毕氏肠微孢子虫在云南部分地区荷斯坦牛中的感染以及基因型分布情况,本实验从云南大理、昆明、曲靖和楚雄等牛养殖集中地区总共9个养殖场采集荷斯坦牛粪便样品547份,采用套式PCR结合测序检测并分析毕氏肠微孢子虫感染情况。结果显示,毕氏肠微孢子虫总感染率为4.57%(25/547);各地区的感染率为0～10.53%,其中昆明地区养殖场感染率最高,不同地区间感染率差异显著(P=0.025<0.05)。断奶前犊牛感染率为6.32%(23/364),高于其他月龄牛的感染率,但各月龄牛感染率差异不显著(P=0.052>0.05)。公牛感染率为7.84%(8/102)高于母牛3.82%(17/445),性别间感染率差异不显著(P=0.079>0.05)。进一步通过对样品扩增产物测序分析后鉴定其基因型,结果显示本研究共鉴定出5个基因型,I、J、BEB4为3个已知的基因型和YNY1、YNY2 2个新拟定的基因型,其中BEB4为优势基因型。在不同地区中,大理检出4种基因型(BEB4、I、YNN1、YNN2),昆明仅检出基因型BEB4,曲靖仅检出基因型J,大理地区基因型分布较为多样;在0...     

马属动物毕氏肠微孢子虫研究进展

毕氏肠微孢子虫是一种引起人和动物腹泻的常见病原体,可通过食物和水源传播,具有重要的公共卫生意义。根据系统进化分析,已鉴定出近300个毕氏肠微孢子虫基因型,分为9个组群,组群1中的基因型大多数具有潜在人兽共患性;组群2~8中的基因型多数具有宿主特异性,部分也可感染人。研究表明,马属动物可感染37个毕氏肠微孢子虫基因型,具有一定人兽共患风险。论文就近年来感染马属动物的毕氏肠微孢子虫基因型、人兽共患风险、流行情况和致病性等进行综述。     

长春地区梅花鹿毕氏肠微孢子虫分子流行病学调查

为了掌握长春地区梅花鹿毕氏肠微孢子虫(Enterocytozoon bieneusi)的感染率及基因型种类的分布情况,探究其在人与动物之间传播的源由,本研究于2018年12月-2019年12月期间采集长春地区8个养殖场共计538份梅花鹿粪便样品进行毕氏肠微孢子虫的流行病学调查及基因型分析,经PCR鉴定,测序分析、序列拼接比对以及系统进化分析。试验结果表明,长春地区梅花鹿毕氏肠微孢子虫的感染率为17.84%(96/538),不同养殖场的感染率为0~47.96%;研究共发现14种基因型(BEB6、EbpC、Ⅰ、JLD-Ⅲ、JLD-Ⅸ、JLD-ⅩⅤ、JLD-ⅩⅥ、JLD-ⅩⅦ、JLD-ⅩⅧ、JLD-ⅩⅨ、JLD-ⅩⅩ、JLD-ⅩⅪ、JLD-ⅩⅫ和JLD-ⅩⅩⅢ),其中BEB6为优势基因型。应用多位点序列分型(MLST)方法,选择MS1、MS3、MS4以及MS7位点对毕氏肠微孢子虫阳性样本进行多态性和群体遗传结构分析,结果显示,在96份阳性样品中,共有16份阳性分离株在至少2个位点同时扩增成功,形成10个多位点基因型(multilocus genotypes,MLGs),相同的基因型之间存在不同亚型,表明长春地区梅花鹿毕氏肠微孢子虫具有丰富的遗传多样性。本研究应用分子生物学方法对长春地区梅花鹿毕氏肠微孢子虫的流行病情况及基因型进行分析,研究结果不仅为梅花鹿毕氏肠微孢子虫防控提供科学依据,也为防控毕氏微孢子虫在人和动物间的传播提供基础数据,对保障公共卫生与健康意义重大。     

毕氏肠微孢子虫基因分型及公共卫生学重要性

微孢子虫(Microsporidia)是一类细胞内专性寄生的真核生物,广泛存在于无脊椎动物如蜜蜂、蚕和几乎所有种类的脊椎动物中.在已知并被命名的1 200多个虫种中,共有8个属中的14个虫种可以感染人,其中毕氏肠微孢子虫(E.bieneusi)、兔脑炎微孢子虫(E.cuniculi)、肠脑炎微孢子虫(E.intestinalis)以及海伦脑炎微孢子虫(E.hellem)较为常见[1-3].E.bieneusi是感染人和动物最常见的病原体,能导致免疫妥协或低下者长期的致死性腹泻,也能够导致免疫功能正常者剧烈的自限性腹泻[2-3].     

南京地区犊牛隐孢子虫感染情况调查

以南京地区３个奶牛场的５５头６月龄以内的犊牛为调查对象，在１６头犊牛的粪便中查到隐孢子虫卵囊，平均感染率为２９．１％（１６／５５）。Ａ、Ｂ２个奶牛场犊牛的感染率（分别是４０．０％和３５．７％）高于Ｃ场犊牛的感染率（１９．２％），但３个奶牛场感染率的统计学差异不显著（Ｐ〉０．０５）。各奶牛场１～３月龄犊牛隐孢子虫的感染强度和感染率均比４～６月龄犊牛高（Ｐ〈０．０５）。     

平安县绵羊住肉孢子虫感染情况调查

住肉孢子虫是重要的人畜共患寄生虫病之一。它主要寄生于牛、羊、马、猪、犬、猫、鼠和人,此外,还寄生于鸟类,鱼类和爬行类等多种动物。在肉食品等公共卫生上影响很大。为了摸清平安县当地绵羊的感染情况,笔者于2001年对平安县牛羊定点屠宰场屠宰的绵羊进行了住肉孢子虫感染情况的调查。现报告如下。     

家蚕肠球菌对微孢子虫体外发芽的抑制作用

肠球菌在家蚕消化道内的高频分布和具有丰富表型的特点 ,使其成为家蚕病害生物防治的良好可利用微生物。本试验从家蚕细菌性肠道病病蚕消化道内分离了甘露醇酵解酶、山梨醇酵解酶和糖元酵解酶表型不同的4个肠球菌菌株。菌株培养上清对家蚕微孢子虫的体外孢子发芽抑制试验表明 :肠球菌对家蚕微孢子虫孢子的发芽具有较强的抑制作用 ,抑制效果的绝对值达 31 4 9%～ 4 3 5 6 % ,相对值达 4 8 18%～ 5 0 5 6 % ,菌株间未显示明显差异     

Cryptosporidium, Giardia and Enterocytozoon bieneusi in cats from Bogota (Colombia) and genotyping of isolates

The prevalence of Cryptosporidium, Giardia, and Enterocytozoon bieneusi in cats from Bogota (Colombia) was determined from fecal specimens and scrapings of duodenal and ileal mucosa screened by PCR. All PCR-positive specimens were sequenced to determine the genotype(s) present. Of 46 cats, 6 (13%) were positive for Cryptosporidium, 5 (11%) were infected with C. felis and one (2%) with C. muris. Three (6.5%) cats were infected with Giardia duodenalis Assemblage F. Eight (17%) cats were infected with four genotypes of E. bieneusi: genotype D-like (9%), K (4%), Peru 10 (2%), and Peru 5 (2%). This is the first report on the presence of zoonotic species/genotypes of Cryptosporidium and E. bieneusi in cats in Colombia.     

Outbreak of microsporidiosis caused by Enterocytozoon bieneusi in falcons

Four falcons from a private collection of 137 falcons in Abu Dhabi (UAE) died suddenly in summer 2005. In order to screen for a possible disease among the remaining falcons in the aviary, all other birds were caught, examined and treated if necessary. Most of the falcons suffered from massive lice infestation and 74 falcons additionally from a heavy Caryospora sp. burden. Endoscopy revealed yellowish plaques on intestines, livers or kidneys in 70 birds (51.1% morbidity). Proliferative serositis was seen in 17 out of 24 necropsied birds with plaques on intestines, livers or kidneys, which did not resemble any known disease in falcons. However, apart from 20 falcons, which died within a 6-week period after the initial examinations due to advanced disease stages, all other falcons responded well to the treatment with dimetridazole (Emtryl), indicating protozoal disease. Immunohistochemistry confirmed the presence of microsporidial antigen. The final diagnosis of Enterocytozoon (E.) bieneusi genotype D was confirmed with materials from 6 birds by PCR and sequencing. To our knowledge this is the first report of microsporidiosis caused by E. bieneusi in raptors in general and in falcons in particular. However, it is still unclear for how long E. bieneusi was present in the falcon flock, and which role it played in the development of the disease. Predisposing factors such as high temperature and overcrowding in the aviary induced immune suppression causing massive lice infestation as well as coccidiosis, thus paving the way for invasion with microsporidial spores.     

First report of Enterocytozoon bieneusi from dairy cattle in Argentina

Fecal specimens were obtained from a total of 70 dairy calves less than two months old on 11 municipalities in Buenos Aires, Argentina. After removal of fecal debris by sieving and sucrose flotation, specimens were subjected to PCR to detect the presence of Enterocytozoon bieneusi. PCR revealed a 14.3% of prevalence for E. bieneusi with 10 positive calves from 7 municipalities. Gene sequence analysis conducted in all samples positives by PCR revealed the presence of six genotypes; four previously reported in cattle as well as humans (D, I, J, and BEB4), one never reported in cattle before but previously reported in humans (EbpC), and one novel genotype (BEB10). These results constitute the first molecular characterization of E. bieneusi in Argentina, and suggest a potential risk of zoonotic transmission in this area.     

First report of Enterocytozoon bieneusi infection on a pig farm in the Czech Republic

Enterocytozoon bieneusi infects humans and animals and can cause life-threatening diarrhea in immunocompromised people. The routes of transmission and its zoonotic potential are not fully understood. Pigs have been frequently reported to have E. bieneusi; therefore, we surveyed farm-raised pigs in the Czech Republic to determine its presence and genetic diversity. Spores were detected by microscopy in the faeces of 65 out of 79 examined animals (82%). A species-specific polymerase chain reaction (PCR) identified E. bieneusi in 94% of samples. Genotyping based on the ITS regions of the SSU rRNA gene identified that most pigs were infected with the species-specific genotype F, while two animals had the zoonotic genotype D and two had genotype Peru 9. This is the first report of E. bieneusi in swine in the Czech Republic, and demonstrated that most infections were with pig-specific genotypes. Nonetheless, swine may still play a role in the transmission of E. bieneusi to humans.     

Cryptosporidium hominis Subtypes and Enterocytozoon bieneusi Genotypes in HIV‐Infected Persons in Ibadan,Nigeria

Cryptosporidium and Enterocytozoon are common opportunistic pathogens in HIV+ patients in developing countries, especially those do not have access to antiretroviral therapy. To determine the distribution of genotypes/subtypes of Cryptosporidium and Enterocytozoon bieneusi, faecal specimens were collected from 132 HIV+ persons attending a tertiary hospital in Ibadan, Nigeria. By polymerase chain reaction, eight and ten patients were identified as positive for Cryptosporidium spp. and E. bieneusi, respectively. Seven of the Cryptosporidium specimens were identified as C. hominis, while the remaining one as the new species C. viatorum recently identified in the United Kingdom. DNA sequencing of the 60‐kDa glycoprotein gene showed that the C. hominis belonged to three common subtype families: Ia (in three patients), Ib (in one patient) and Ie (in one patient). In contrast, DNA sequencing of the E. bieneusi internal transcribed spacer products showed the occurrence of genotypes associated with both humans (Peru 8 in one patient, Nig2 in two patients and a new genotype in one patient) and animals (D in one patient and Type IV in five patients). Low CD4+ cell count was identified as a risk factor for both cryptosporidiosis and microsporidiosis.     

Longitudinal identification of Enterocytozoon bieneusi in dairy calves on a farm in Southern Xinjiang,China

Enterocytozoon bieneusi is the most common species responsible for human and animals microsporidiasis. A total of 250 samples were collected weekly from 25 newborn dairy calves of a farm in Southern Xinjiang, China at one to ten weeks of age. Enterocytozoon bieneusi was identified and genotyped by nested PCR amplification and sequencing of internal transcribed spacer (ITS) region.The cumulative prevalence of E. bieneusi infection was 100% (25/25), and the average infection was 52.0% (130/250). The highest infection rate was recorded at six weeks of age (92.0%, 23/25), and no infection was observed at one and two weeks of age. Sequencing analysis showed nine E. bieneusi genotypes (J, EbpC, PigEBITS5, CHV4, CHC3, CS-9, KIN-1, CH5, and CAM5) were identified. The highest genetic polymorphism was observed at ten weeks of age. Genotype J was the predominant E. bieneusi genotype. Phylogenetic analysis clustered genotype J into Group 2 and other eight genotypes (EbpC, PigEBITS5, CHV4, CHC3, CS-9, KIN-1, CH5, and CAM5), detected in 22 (16.9%, 22/130) samples, into Group 1. Among the genotypes, EbpC, KIN-1, and J have been identified in humans. The highest E. bieneusi infection rate (57.9%, 124/214) was observed in fecal samples with formed feces with no diarrhea (p < 0.01), and high genetic polymorphism was observed in class I fecal samples. The presence of zoonotic E. bieneusi genotypes in dairy calves suggests the possibility of transmitting zoonotic infections to humans. It provides the basic data on dynamic change of E. bieneusi in calves.