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Quantification of hookworm ova from wastewater matrices using quantitative PCR.

Pradip Gyawali1, Warish Ahmed2, Jatinder P Sidhu1

  • 1CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Qld 4102, Australia; School of Public Health, The University of Queensland, Herston Road, Qld 4006, Australia.

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|June 26, 2017
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Summary

A new quantitative PCR (qPCR) assay accurately quantifies hookworm (Ancylostoma caninum) ova in wastewater and sludge. This method provides reliable detection for environmental monitoring and public health assessments.

Keywords:
Health risksHookworm ovaQuantitative PCRWastewater matrices

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Area of Science:

  • Environmental microbiology
  • Molecular parasitology
  • Public health

Background:

  • Wastewater and sludge can harbor parasitic ova, posing public health risks.
  • Accurate quantification of parasitic ova in environmental samples is crucial for risk assessment.
  • Existing methods for ova detection may lack sensitivity or specificity.

Purpose of the Study:

  • To develop and validate a quantitative PCR (qPCR) assay for detecting Ancylostoma caninum ova.
  • To estimate the number of hookworm ova in wastewater and sludge samples.
  • To assess the utility of qPCR for environmental monitoring of parasitic contamination.

Main Methods:

  • Development of a novel qPCR assay targeting Ancylostoma caninum.
  • Estimation of average gene copy numbers per ovum using a mixed ova population.
  • Validation of the qPCR assay by seeding known quantities of ova into wastewater and sludge.
  • Application of the qPCR assay to analyze treated wastewater, raw wastewater, and sludge samples.

Main Results:

  • The qPCR assay estimated an average of 3.7×10^3 gene copies per ovum.
  • The assay demonstrated good accuracy in quantifying seeded hookworm ova in treated wastewater.
  • Ancylostoma caninum gene copies were detected in 50% of treated wastewater, 90% of raw wastewater, and 67% of sludge samples.
  • Average ova counts were estimated at 0.02 in treated wastewater, 1.24 in raw wastewater, and 67 in sludge.

Conclusions:

  • Quantitative PCR is a viable method for quantifying hookworm ova in wastewater and sludge.
  • The developed qPCR assay offers a sensitive tool for detecting parasitic contamination in environmental matrices.
  • Caution is recommended when interpreting qPCR data for health risk assessments due to potential variability.