de Lucio Aida, Amor-Aramendía Aranzazu, Bailo Begoña, Saugar José M, Anegagrie Melaku, Arroyo Ana, López-Quintana Beatriz, Zewdie Derjew, Ayehubizu Zimmam, Yizengaw Endalew, Abera Bayeh, Yimer Mulat, Mulu Wondemagen, Hailu Tadesse, Herrador Zaida, Fuentes Isabel, Carmena David
Parasitology Service, National Centre for Microbiology, Majadahonda, Madrid, Spain.
National Centre of Tropical Medicine, Madrid, Spain.
PLoS One. 2016 Jul 28;11(7):e0159992. doi: 10.1371/journal.pone.0159992. eCollection 2016.
Giardia duodenalis and Cryptosporidium spp. are enteric protozoan causing gastrointestinal illness in humans and animals. Giardiasis and cryptosporidiosis are not formally considered as neglected tropical diseases, but belong to the group of poverty-related infectious diseases that impair the development and socio-economic potential of infected individuals in developing countries.
We report here the prevalence and genetic diversity of G. duodenalis and Cryptosporidium spp. in children attending rural primary schools in the Bahir Dar district of the Amhara Region, Ethiopia. Stool samples were collected from 393 children and analysed by molecular methods. G. duodenalis was detected by real-time PCR, and the assemblages and sub-assemblages were determined by multilocus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Detection and identification of Cryptosporidium species was carried out by sequencing of a partial fragment of the small-subunit ribosomal RNA gene.
The PCR-based prevalences of G. duodenalis and Cryptosporidium spp. were 55.0% (216/393) and 4.6% (18/393), respectively. A total of 78 G. duodenalis isolates were successfully characterized, revealing the presence of sub-assemblages AII (10.3%), BIII (28.2%), and BIV (32.0%). Discordant typing results AII/AIII and BIII/BIV were identified in 7.7% and 15.4% of the isolates, respectively. An additional five (6.4%) isolates were assigned to assemblage B. No mixed infections of assemblages A+B were found. Extensive genetic variation at the nucleotide level was observed within assemblage B (but no within assemblage A), resulting in the identification of a large number of sub-types. Cryptosporidium diversity was demonstrated by the occurrence of C. hominis, C. parvum, and C. viatorum in the population under study.
Our data suggest an epidemiological scenario with an elevated transmission intensity of a wide range of G. duodenalis genetic variants. Importantly, the elevated degree of genetic diversity observed within assemblage B is consistent with the occurrence of intra-assemblage recombination in G. duodenalis.
十二指肠贾第虫和隐孢子虫属是引起人和动物胃肠道疾病的肠道原生动物。贾第虫病和隐孢子虫病虽未被正式视为被忽视的热带疾病,但属于与贫困相关的传染病,会损害发展中国家受感染个体的发育和社会经济潜力。
我们在此报告埃塞俄比亚阿姆哈拉州巴赫达尔地区农村小学儿童中十二指肠贾第虫和隐孢子虫属的流行情况及遗传多样性。从393名儿童中采集粪便样本并采用分子方法进行分析。通过实时荧光定量PCR检测十二指肠贾第虫,并通过对该寄生虫的谷氨酸脱氢酶和β-贾第蛋白基因进行多位点序列基因分型来确定其组合和亚组合。通过对小亚基核糖体RNA基因的部分片段进行测序来检测和鉴定隐孢子虫种类。
基于PCR的十二指肠贾第虫和隐孢子虫属的流行率分别为55.0%(216/393)和4.6%(18/393)。共成功鉴定了78株十二指肠贾第虫分离株,发现存在亚组合AII(10.3%)、BIII(28.2%)和BIV(32.0%)。分别在7.7%和15.4%的分离株中鉴定出不一致的分型结果AII/AIII和BIII/BIV。另外5株(6.4%)分离株被归为组合B。未发现组合A+B的混合感染。在组合B内观察到核苷酸水平的广泛遗传变异(但组合A内未观察到),从而鉴定出大量亚型。在所研究的人群中发现了人隐孢子虫、微小隐孢子虫和旅行者隐孢子虫,证明了隐孢子虫的多样性。
我们的数据表明存在一种流行病学情况,即多种十二指肠贾第虫遗传变异体的传播强度升高。重要的是,在组合B内观察到的高度遗传多样性与十二指肠贾第虫组合内重组的发生一致。