Quantitative detection of viable helminth ova from raw wastewater, human feces, and environmental soil samples using novel PMA-qPCR methods

Gyawali, P., Ahmed, W., Sidhu, J. P. S., Nery, S. V., Clements, A. C., Traub, R., McCarthy, J. S., Llewellyn, S., Jagals, P. and Toze, S. (2016) Quantitative detection of viable helminth ova from raw wastewater, human feces, and environmental soil samples using novel PMA-qPCR methods. Environmental Science and Pollution Research, 23 18: 18639-18648. doi:10.1007/s11356-016-7039-9


Author Gyawali, P.
Ahmed, W.
Sidhu, J. P. S.
Nery, S. V.
Clements, A. C.
Traub, R.
McCarthy, J. S.
Llewellyn, S.
Jagals, P.
Toze, S.
Title Quantitative detection of viable helminth ova from raw wastewater, human feces, and environmental soil samples using novel PMA-qPCR methods
Journal name Environmental Science and Pollution Research   Check publisher's open access policy
ISSN 0944-1344
1614-7499
Publication date 2016-09
Year available 2016
Sub-type Article (original research)
DOI 10.1007/s11356-016-7039-9
Open Access Status Not yet assessed
Volume 23
Issue 18
Start page 18639
End page 18648
Total pages 10
Place of publication Heidelberg, Germany
Publisher Springer
Collection year 2017
Language eng
Formatted abstract
In this study, we have evaluated the efficacy of propidium monoazide quantitative polymerase chain reaction (PMA-qPCR) to differentiate between viable and non-viable Ancylostoma caninum ova. The newly developed method was validated using raw wastewater seeded with known numbers of A. caninum ova. Results of this study confirmed that PMA-qPCR has resulted in average of 88 % reduction (P < 0.05) in gene copy numbers for 50 % viable +50 % non-viable when compared with 100 % viable ova. A reduction of 100 % in gene copies was observed for 100 % non-viable ova when compared with 100 % viable ova. Similar reductions (79–80 %) in gene copies were observed for A. caninum ova-seeded raw wastewater samples (n = 18) collected from wastewater treatment plants (WWTPs) A and B. The newly developed PMA-qPCR method was applied to determine the viable ova of different helminths (A. caninum, A. duodenale, Necator americanus and Ascaris lumbricoides) in raw wastewater, human fecal and soil samples. None of the unseeded wastewater samples were positive for the above-mentioned helminths. N. americanus and A. lumbricoides ova were found in unseeded human fecal and soil samples. For the unseeded human fecal samples (1 g), an average gene copy concentration obtained from qPCR and PMA-qPCR was found to be similar (6.8 × 105 ± 6.4 × 105 and 6.3 × 105 ± 4.7 × 105) indicating the presence of viable N. americanus ova. Among the 24 unseeded soil samples tested, only one was positive for A. lumbricoides. The mean gene copy concentration in the positively identified soil sample was 1.0 × 105 ± 1.5 × 104 (determined by qPCR) compared to 4.9 × 104 ± 3.7 × 103 (determined by PMA-qPCR). The newly developed PMA-qPCR methods were able to detect viable helminth ova from wastewater and soil samples and could be adapted for health risk assessment.
Keyword Hookworm ova
Viability
Wastewater
PMA-qPCR
Public health
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

 
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Created: Mon, 11 Jul 2016, 17:36:05 EST by Paul Jagals on behalf of School of Public Health