Date of Award

2019

Document Type

Open Access Master's Thesis

Degree Name

Master of Science in Biological Sciences (MS)

Administrative Home Department

Department of Biological Sciences

Advisor 1

Karyn Fay

Advisor 2

Jennifer Becker

Committee Member 1

Eric Seagren

Abstract

The current method for detection of Ascaris ova in biosolids and wastewater has several drawbacks including being labor and time intensive as well as being difficult to quantify viable ova that have larvated during treatment. The goal of this thesis was to improve several methods for the detection and quantification of Ascaris ova during long term-storage of biosolids including improvement of quantifying viable ova using a heat and bleach treatment to promote movement of larvae inside of ova, and comparison of the traditional microscopic method with qPCR and PMA-qPCR to prevent the false-positive results. Improved methods to promote movement of larvated ova using bleach and heat treatment were tested and shown to increase ova movement when compared with no treatment or only heat or bleach treatment, thus decreasing overall count time. qPCR and the traditional microscopic method were tested using long-term stored biosolids, which when compared showed a correlation between microscopically counted and qPCR calculated ova. PMA-qPCR was tested as a way to prevent false positives from inactivated but not yet degraded larval ova, however no decrease in amplification was observed for PMA treated samples when compared to non-treated samples. qPCR may be a valid method for quantifying Ascaris ova during different long-term treatment options and evaluating the safety of biosolids.

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