Asymptomatic carriage of intestinal protists is common in children in Lusaka Province, Zambia

PLoS Negl Trop Dis. 2024 Dec 13;18(12):e0012717. doi: 10.1371/journal.pntd.0012717. eCollection 2024 Dec.

Abstract

Background: PCR-based screenings on the presence of diarrhoea-causing intestinal protist species are limited in Zambia, resulting in inaccurate current prevalence and epidemiological data. Sensitive PCR-based methods are particularly well suited for detecting subclinical infections in apparently healthy carriers.

Methodology: In this prospective cross-sectional study, we investigated the occurrence of the most common intestinal protists in an apparently healthy paediatric population (5-18 years) in Lusaka Province, Zambia. We collected single stool samples (n = 256) and epidemiological questionnaires on demographics, behavioural habits, drinking water and toilet access from participating children. We used PCR for the initial screening of samples for the presence of intestinal protist species and Sanger and next-generation sequencing for genotyping. We conducted statistical analyses to assess the association of the gathered variables with an increased likelihood of the investigated pathogens.

Principal findings: Blastocystis sp. was the most prevalent intestinal protist found (37.9%, 97/256; 95% CI: 31.9-44.1), followed by Giardia duodenalis (30.9%, 79/256; 95% CI: 25.3-36.90), Entamoeba dispar (13.3%, 34/256; 95% CI: 9.4-18.1), and Cryptosporidium spp. (4.3%, 11/256, 95% CI: 2.2-7.6). Entamoeba histolytica was not detected. Based on Sanger sequencing results, subtypes ST2 (44.3%, 43/97), ST1 (35.1%, 34/97), and ST3 (20.6%, 20/97) were identified within Blastocystis sp. and assemblages B (71.0%), A+B (16.1%), and A (12.9%) within G. duodenalis. Cryptosporidium parvum (81.8%) and C. hominis (18.2%) were the only two Cryptosporidium species found. Living in the Kafue District was positively associated with higher infection rates by G. duodenalis and Blastocystis sp. Schoolchildren living in Chongwe District were more likely to be infected by Cryptosporidium spp.

Conclusions/significance: Intestinal protist infection/colonization is a common finding in apparently healthy children in Lusaka Province, Zambia. Asymptomatic carriers may play an underestimated role as spreaders of gastrointestinal parasitic infections. This study improves our current understanding of the epidemiology of diarrhoea-causing protists in Zambia and sub-Saharan Africa and indicates that the role of asymptomatic carriers of gastrointestinal parasites in transmission should be further explored.

MeSH terms

  • Adolescent
  • Asymptomatic Infections / epidemiology
  • Blastocystis / classification
  • Blastocystis / genetics
  • Blastocystis / isolation & purification
  • Carrier State* / epidemiology
  • Carrier State* / parasitology
  • Child
  • Child, Preschool
  • Cross-Sectional Studies
  • Cryptosporidium / classification
  • Cryptosporidium / genetics
  • Cryptosporidium / isolation & purification
  • Entamoeba / classification
  • Entamoeba / genetics
  • Entamoeba / isolation & purification
  • Feces / parasitology
  • Female
  • Giardia lamblia* / genetics
  • Giardia lamblia* / isolation & purification
  • Humans
  • Intestinal Diseases, Parasitic / epidemiology
  • Intestinal Diseases, Parasitic / parasitology
  • Male
  • Polymerase Chain Reaction
  • Prevalence
  • Prospective Studies
  • Zambia / epidemiology

Grants and funding

This study was partially funded by the Health Institute Carlos III, Spanish Ministry of Economy and Competitiveness, (PI16CIII/00024 to DC) and by the USDA-ARS (8042-32000-112-00-D to MS). The Article Processing Charge for this publication was covered by Women for Africa Foundation and Eurofins. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.