Abstract
Accurate evaluation of viable Ascaris ova in wastewater is the key to mitigating Ascaris reinfections in endemic regions. In this study, the viability of Ascaris ova in raw wastewater was determined using three different detection methods: culture-based, BacLight Live/Dead staining and propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR). Furthermore, comparative assessment of viability utilising the aforementioned detection methods was performed using seeded experiments in wastewater. The percentage of viability was: culture-based (82%), BacLight Live/Dead staining (87%) and PMA-qPCR (85%) respectively. Despite the fact that no statistical difference was shown in the viability determination among the three methods, PMA-qPCR-based viability determination would be preferable over the other two methods for evaluating potential public health risks with A. suum ova due to its accuracy, being least subjective and its rapid reaction time.
Highlights
Wastewater use is expected to increase globally to meet rapidly growing demands for potable and non-potable water supplies due to growing population and changing climate (Hanjra et al )
Viability determination of A. suum ova recovered from pig faeces using the BacLight Live/Dead staining and observation by confocal laser scanning microscopy (CLSM) showed that the inner lipoprotein layer of most of the ova was stained with both Syto 9 and PI; the internal structures were unstained, denoting that these ova were viable
The mean percentage of viability determined using the BacLight live/dead staining method for untreated ova in raw wastewater was 87%, slightly higher, not statistically significant (Fisher exact test, p > 0.05), than the viability determined using either the culture-based method (82%) or Propidium monoazide (PMA)-quantitative polymerase chain reaction (qPCR) (85%) (Table 2)
Summary
Wastewater use is expected to increase globally to meet rapidly growing demands for potable and non-potable water supplies due to growing population and changing climate (Hanjra et al ). It has been estimated that around 20 million ha of agricultural lands are irrigated with treated as well as raw wastewater (Jiménez & Asano ). Direct application of raw wastewater into agricultural land is a common practice in developing countries, an activity that is rarely reported. Wastewater represents a reusable water source; yet it has significant potential to act as a reservoir for waterborne disease transmission (Sidhu & Toze ; Li et al ).
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