Longitudinal assessment of the exposure to Ascaris lumbricoides through copromicroscopy and serology in school children from Jimma Town, Ethiopia

Background We previously demonstrated that serology holds promise as an alternative diagnostic tool to copromicroscopy to monitor and evaluate deworming programs targeting soil-transmitted helminths (STHs). Here we explored the dynamics of anti-Ascaris antibodies (Ab) and evaluated the Ab-isotype of choice to assess the longitudinal exposure to Ascaris in Ethiopian school children. Methodology Between October 2018 and February 2020, stool and blood samples were collected every four months from school children (4 to 6 years of age). Stool samples were analyzed by duplicate Kato-Katz to assess the presence and intensity of any STH infection. Plasma Ab-responses against the total extract of Ascaris suum lung third stage larvae were measured through in-house Ab-ELISA's for seven different Ab-isotypes. Principal findings At baseline, 42.4% of the 66 children were excreting eggs of any STH, Trichuris (37.9%) being the most prevalent. The cumulative prevalence (proportion of children tested that positive at least once over the entire study period) was 56.1% for Trichuris and 31.8% for Ascaris. For Ascaris, re-infections were frequently observed, whereas for Trichuris, children often remained excreting eggs following drug administration. When measuring anti-Ascaris Ab-levels, the cumulative seroprevalence was generally higher (IgG4: 60.6%; IgG1: 50.0%; IgE: 36.4%). The individual anti-Ascaris IgG4 levels at baseline were positively associated with the fecal egg counts averaged over the study period, the rate of egg-appearance and the number of positive test results. There was no apparent cross-reactivity between the anti-Ascaris IgG4 Ab-ELISA and Trichuris. Conclusions/Significance We demonstrate that the children are exposed to STH before the age of four and that both the exposure to Ascaris, highlighting that the exposure to disease is underestimated when measured with copromicroscopy. Compared to other Ab-isotypes, IgG4 is the Ab-isotype of choice to measure Ascaris exposure in STH endemic settings. Finally, the results also highlight that measuring anti-Ascaris IgG4 levels holds promise as a tool to identify individuals at higher risk for continued exposure to this STH. Author summary World-wide large-scale deworming programs are being implemented to control the morbidity attributable to intestinal worms. Our group has recently shown that diagnostic tools that detect anti-parasite antibodies hold promise as an alternative to the current diagnostic standard (detecting worm eggs in stool using microscopy) to monitor and evaluate the progress of these programs towards the set targets. In the present study, we further explored the potential of antibody-based assays by monitoring the exposure of Ethiopian school children during a 17-month period to worm infections. Our results indicated that children in this area are exposed to intestinal worms from an early age onwards and that, based on serological results, more children are exposed to worm infections than measured by the current diagnostic standard. In addition, we demonstrated that test results of the antibody assay at the start of the study were positively associated with mean egg counts averaged over the study period, the rate of egg-appearance and the number of positive stool tests, highlighting that such assay holds promise as a tool to identify individuals at higher high risk for continued exposure to worms, and ultimately improve control programs.