Genotypic characterization of Giardia lamblia, Entamoeba species and Cryptosporidium species among children in Kiambu County, Kenya

IF 2.9 Q2 PARASITOLOGY

Food and Waterborne Parasitology Pub Date : 2025-05-11 DOI:10.1016/j.fawpar.2025.e00268

Liza Kiende Mwirigi , Cecilia Mbae , Margaret Muturi , Scholastica Mathenge , Michael Mugo , Tabitha Irungu , Benjamin Ngugi , Erastus Mulinge

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Abstract

Enteric parasitic infections are a leading cause of diarrheal disease and malnutrition in low- and middle-income countries like Kenya. Among the most prevalent protozoan pathogens in children are Entamoeba histolytica, Giardia lamblia, and Cryptosporidium species. However, data on their genetic diversity, co-occurrence, and variability in Kenya remain limited. This study investigated the genetic diversity of G. lamblia, Entamoeba spp., and Cryptosporidium spp. in children aged 10 years and below in Kiambu County, Kenya. A total of 550 stool samples were analyzed for enteric parasites using formal-ether concentration and Modified Ziehl-Neelsen staining. Genomic DNA was extracted from microscopy-positive samples, and species-specific nested polymerase chain reaction was performed to genotype Entamoeba spp. using the 18S ribosomal RNA gene. For G. lamblia and Cryptosporidium spp., nested PCR and sequencing targeted the βeta-giardin, triose phosphate isomerase, and 60-kDa glycoprotein genes, respectively. Microscopy detected Entamoeba spp. (29.6 %, 163/550), G. lamblia (14.6 %, 80/550), and Cryptosporidium spp. (1.3 %, 7/550). PCR analysis identified E. histolytica (3.3 %, 18/550), E. dispar (3.8 %, 21/550), E. moshkovskii (1.6 %, 9/550), E. coli (13.1 %, 72/550), and E. hartmanni (1.5 %, 8/550). Sequence analysis of the tpi and β-giardin genes identified G. lamblia assemblages A (20/50) and B (30/50). All assemblage A isolates were classified as sub-assemblage AII (20/20), while assemblage B isolates were further subdivided into sub-assemblages BIII (21/30) and BIV (9/30). All Cryptosporidium isolates were identified as C. hominis, with subtypes IbA9G3 (5/6) and IeA11G3T3 (1/6). Microscopy results revealed a significant association between Entamoeba spp. and Cryptosporidium spp. with both age groups and study sites. Entamoeba dispar by PCR and G. lamblia by microscopy showed significant differences between study sites. Additionally, the distribution of G. lamblia assemblages A and B, along with sub-assemblages AII, BIII, and BIV, differed significantly between the study sites. Among these, only sub-assemblage BIV showed a significant association with age groups. The detection of E. histolytica alongside related Entamoeba spp. underscores the importance of molecular diagnostics for accurate amoebiasis management and epidemiological surveillance. Additionally, the identification of G. lamblia sub-assemblages AII, BIII, and BIV, as well as C. hominis subtypes, suggests anthroponotic transmission, emphasizing the need for improved sanitation and public health interventions.

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肯尼亚Kiambu县儿童中贾第鞭毛虫、内阿米巴原虫和隐孢子虫的基因型特征

肠道寄生虫感染是肯尼亚等低收入和中等收入国家腹泻和营养不良的主要原因。在儿童中最常见的原生动物病原体是溶组织内阿米巴原虫、贾第鞭毛虫和隐孢子虫。然而，关于它们在肯尼亚的遗传多样性、共现性和变异性的数据仍然有限。本研究调查了肯尼亚Kiambu县10岁及以下儿童中兰螺旋体、内阿米巴原虫和隐孢子虫的遗传多样性。采用甲醛乙醚浓度法和改良Ziehl-Neelsen染色法对550份粪便样本进行肠道寄生虫分析。镜检阳性标本提取基因组DNA，利用18S核糖体RNA基因对内阿米巴原虫进行巢式聚合酶链反应分型。对于蓝氏螺旋体和隐孢子虫，巢式PCR和测序分别针对β -贾丁素、磷酸三糖异构酶和60 kda糖蛋白基因。镜检检出内阿米巴原虫（29.6%,163/550）、兰螺旋体（14.6%,80/550）和隐孢子虫（1.3%,7/550）。PCR分析鉴定出溶组织大肠杆菌（3.3%,18/550）、dispar大肠杆菌（3.8%,21/550）、moshkovskii大肠杆菌（1.6%,9/550）、大肠杆菌（13.1%,72/550）和hartmanni大肠杆菌（1.5%,8/550）。对tpi和β-贾丁素基因进行序列分析，鉴定出兰氏螺旋体A（20/50）和B（30/50）组合。组合A分离株被划分为亚组合AII(20/20)，组合B分离株被进一步划分为亚组合BIII（21/30）和BIV（9/30）。所有隐孢子虫分离株均鉴定为人隐孢子虫，亚型为IbA9G3（5/6）和IeA11G3T3（1/6）。显微镜检查结果显示，内阿米巴和隐孢子虫在年龄组和研究地点都有显著的相关性。PCR法鉴定的异内阿米巴和显微镜法鉴定的兰氏格氏虫在不同研究点间差异显著。此外，在不同的研究地点，G. lamblia组合A和B以及亚组合AII、BIII和BIV的分布也存在显著差异。其中，只有亚组合BIV与年龄组有显著的相关性。溶组织芽胞杆菌与相关内阿米巴原虫的检测强调了分子诊断对阿米巴病准确管理和流行病学监测的重要性。此外，鉴定出的G. lambla亚组合AII、BIII和BIV以及人原C.亚型表明存在人传传播，强调了改善卫生条件和公共卫生干预措施的必要性。

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来源期刊

Food and Waterborne Parasitology Immunology and Microbiology-Parasitology

CiteScore

5.10

自引率

4.00%

发文量

审稿时长

13 weeks

期刊介绍： Food and Waterborne Parasitology publishes high quality papers containing original research findings, investigative reports, and scientific proceedings on parasites which are transmitted to humans via the consumption of food or water. The relevant parasites include protozoa, nematodes, cestodes and trematodes which are transmitted by food or water and capable of infecting humans. Pertinent food includes products of animal or plant origin which are domestic or wild, and consumed by humans. Animals and plants from both terrestrial and aquatic sources are included, as well as studies related to potable and other types of water which serve to harbor, perpetuate or disseminate food and waterborne parasites. Studies dealing with prevalence, transmission, epidemiology, risk assessment and mitigation, including control measures and test methodologies for parasites in food and water are of particular interest. Evidence of the emergence of such parasites and interactions among domestic animals, wildlife and humans are of interest. The impact of parasites on the health and welfare of humans is viewed as very important and within scope of the journal. Manuscripts with scientifically generated information on associations between food and waterborne parasitic diseases and lifestyle, culture and economies are also welcome. Studies involving animal experiments must meet the International Guiding Principles for Biomedical Research Involving Animals as issued by the Council for International Organizations of Medical Sciences.