Schistosoma species detection by environmental DNA assays in African freshwaters

PLoS Negl Trop Dis. 2020 Mar 23;14(3):e0008129. doi: 10.1371/journal.pntd.0008129. eCollection 2020 Mar.

Abstract

Background: Schistosomiasis is a neglected tropical parasitic disease associated with severe pathology, mortality and economic loss worldwide. Programs for disease control may benefit from specific and sensitive diagnostic methods to detect Schistosoma trematodes in aquatic environments. Here we report the development of novel environmental DNA (eDNA) qPCR assays for the presence of the human-infecting species Schistosoma mansoni, S. haematobium and S. japonicum.

Methodology/principal findings: We first tested the specificity of the assays across the three species using genomic DNA preparations which showed successful amplification of target sequences with no cross amplification between the three focal species. In addition, we evaluated the specificity of the assays using synthetic DNA of multiple Schistosoma species, and demonstrated a high overall specificity; however, S. japonicum and S. haematobium assays showed cross-species amplification with very closely-related species. We next tested the effectiveness of the S. mansoni assay using eDNA samples from aquaria containing infected host gastropods, with the target species revealed as present in all infected aquaria. Finally, we evaluated the effectiveness of the S. mansoni and S. haematobium assays using eDNA samples from eight discrete natural freshwater sites in Tanzania, and demonstrated strong correspondence between infection status established using eDNA and conventional assays of parasite prevalence in host snails.

Conclusions/significance: Collectively, our results suggest that eDNA monitoring is able to detect schistosomes in freshwater bodies, but refinement of the field sampling, storage and assay methods are likely to optimise its performance. We anticipate that environmental DNA-based approaches will help to inform epidemiological studies and contribute to efforts to control and eliminate schistosomiasis in endemic areas.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Environmental / isolation & purification*
  • DNA, Helminth / isolation & purification
  • Environmental Monitoring
  • Fresh Water / parasitology*
  • Genes, Helminth / genetics
  • Nucleic Acid Amplification Techniques / veterinary
  • Phylogeny
  • Real-Time Polymerase Chain Reaction / veterinary
  • Schistosoma / classification*
  • Schistosoma / genetics*
  • Schistosoma / isolation & purification*
  • Schistosoma haematobium / genetics
  • Schistosoma haematobium / isolation & purification
  • Schistosoma japonicum / genetics
  • Schistosoma japonicum / isolation & purification
  • Schistosoma mansoni / genetics
  • Schistosoma mansoni / isolation & purification
  • Schistosomiasis / epidemiology
  • Schistosomiasis / parasitology
  • Schistosomiasis mansoni / epidemiology
  • Schistosomiasis mansoni / parasitology
  • Snails / parasitology
  • Species Specificity
  • Tanzania

Substances

  • DNA, Environmental
  • DNA, Helminth

Grants and funding

HA was funded by the Embassy of the Kingdom of Saudi Arabia and Prince Nourah Bin Abdulrahman University. MJG and BPN were supported by the Royal Society-Leverhulme Trust Africa Award AA130107. MJG and RAC were supported by Global Challenges Research Fund pump priming project “Aquatic environmental DNA for enhanced health and food security in Tanzania.” The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.