Neglected tropical diseases (NTDs) cause over one million deaths each year; the majority of which are caused by diseases such as Chagas disease, African trypanosomiasis, intestinal and nematode infections, such as giardiasis, Schistosomiases, and many others. The treatment options are not effective in treating all stages of the parasitic life cycle or the phase of the disease. Therefore, a novel approach was used to investigate human milk oligosaccharides (HMOs), which are complex carbohydrate found in breast milk, on parasitic organisms because of their role as anti-adhesive antimicrobials that have proven to lower the risk of pathogenic infections in infants. This study was designed to gain insight on the effect of structurally diverse HMOs on different parasites including Naegleria fowleri, Giardia lamblia, Trypanosoma cruzi, Trypanosoma brucei, and Schistosoma. Through whole organism screenings, purified pooled HMOs from donor samples and synthetic individual HMOs were tested against each of the parasites. The results obtained did not demonstrate a reproducible inhibitory or faciliatory effect on T. brucei, T. cruzi, G. lamblia and N. fowleri. However, a mild phenotypic change in Schistosoma mansoni was observed post incubation with the pooled HMO. In conclusion, the results obtained demonstrate that HMOs (pooled and individual) did not inhibit or enhance the growth of any of the protozoa. Potentially, an alternative experimental approach will be used to investigate the antiadhesive property of HMOs on the pathogenesis of G. lamblia in a co-culture experiment with gastrointestinal tract epithelial cells.