A modified approach to recover and enumerate Ascaris ova in wastewater and sludge

Ravindran, Vivek B.; Surapaneni, Aravind; Crosbie, Nicholas D.; Schmidt, Jonathan; Shahsavari, Esmaeil; Haleyur, Nagalakshmi; Soni, Sarvesh K.; Ball, Andrew S.

doi:10.1371/journal.pntd.0007020

Cited by 8 publications

(14 citation statements)

References 16 publications

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“…Ascaris lumbricoides and Ascaris suum eggs were donated by Helen Williams from the School of Science, RMIT University. They were purified by the zinc sulphate concentration method 58 .…”

Section: Methodsmentioning

confidence: 99%

Real-time detection and identification of nematode eggs genus and species through optical imaging

Qazi

Khalid

Poddar

et al. 2020

Sci Rep

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Nematode eggs are pervasive pathogens that infect billions of people and livestock every year. Adult parasitic nematode worms can be distinguished based on their size and morphology. However, their eggs, particularly their species Ascaris lumbricoides and Ascaris suum cannot be identified from each other. Identifying eggs of helminths from wastewater and sludge is important from a public health perspective to minimize the spread of Ascaris infections. Numerous methods exist for nematode identification, from a morphological-based approach to high throughput sequencing technology. However, these techniques are not consistent and often laborious and time-consuming. In this study, we demonstrate that non-invasive real-time identification of eggs is possible based on their intrinsic fluorescence. Using confocal microscopy, we investigate the autofluorescence properties of five species of nematode eggs and observe clear differences between genus and for the first time their species in sludge samples. This non-invasive imaging technique could lead to better understanding of these species and may assist in early control of diseases.

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“…Ascaris lumbricoides and Ascaris suum eggs were donated by Helen Williams from the School of Science, RMIT University. They were purified by the zinc sulphate concentration method 58 .…”

Section: Methodsmentioning

confidence: 99%

Real-time detection and identification of nematode eggs genus and species through optical imaging

Qazi

Khalid

Poddar

et al. 2020

Sci Rep

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“…Despite high standards of urban wastewater treatment and good sanitation systems in developed countries, the removal of Ascaris ova from wastewater remains crucial to allow the safe use of recycled water in agriculture [7]. Conventional monitoring for helminth ova in these environments relies on culture-based methods such as incubation and optical microscopy, which have many limitations, such as being time-consuming (up to 4 weeks) and laborious which hinder their usefulness as a detection tool [8].…”

Section: Methods Detailsmentioning

confidence: 99%

“…Conventional monitoring for helminth ova in these environments relies on culture-based methods such as incubation and optical microscopy, which have many limitations, such as being time-consuming (up to 4 weeks) and laborious which hinder their usefulness as a detection tool [8]. Despite the advancement in molecular techniques, the need for sophisticated devices is a major constraint, especially in endemic and resource limited settings where reliable means of power supply may not be available [7].…”

Section: Methods Detailsmentioning

confidence: 99%

Detection of helminth ova genera using in-situ biosynthesis of gold nanoparticles

Ravindran¹,

Truskewycz²,

Ball³

et al. 2019

MethodsX

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“…The A. suum infected pig faecal samples were obtained from an abattoir in Laverton, Australia. The ova of A. suum were recovered using a modified version of the Tulane method [21,52]. Aliquots of pig faecal samples with approximately 5 g DS (dissolved solid) were rinsed with milliQ water, homogenised for 1 min with a blender, and allowed to settle in 1% 7X detergent for 30 min (MP Bio, Australia).…”

Section: Source Of Ascaris Ovamentioning

confidence: 99%

“…Faecal samples were collected from pigs that were infected with T. suis. The ova were recovered using Tulane method with minor modifications (21,52) and were also used for further experiments. Genomic DNA of A. lumbricoides and H contortus were obtained from the Faculty of Veterinary and Agricultural Sciences, The University of Melbourne.…”

Section: Detection Limit and Specificitymentioning

confidence: 99%

Detection of Helminth Ova in Wastewater Using Recombinase Polymerase Amplification Coupled to Lateral Flow Strips

Ravindran

Khallaf

Surapaneni

et al. 2020

Water

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Ascaris lumbricoides is a major soil-transmitted helminth that is highly infective to humans. The ova of A. lumbricoides are able to survive wastewater treatment, thus making it an indicator organism for effective water treatment and sanitation. Hence, Ascaris ova must be removed from wastewater matrices for the safe use of recycled water. Current microscopic techniques for identification and enumeration of Ascaris ova are laborious and cumbersome. Polymerase chain reaction (PCR)-based techniques are sensitive and specific, however, major constraints lie in having to transport samples to a centralised laboratory, the requirement for sophisticated instrumentation and skilled personnel. To address this issue, a rapid, highly specific, sensitive, and affordable method for the detection of helminth ova was developed utilising recombinase polymerase amplification (RPA) coupled with lateral flow (LF) strips. In this study, Ascaris suum ova were used to demonstrate the potential use of the RPA-LF assay. The method was faster (< 30 min) with optimal temperature at 37 °C and greater sensitivity than PCR-based approaches with detection as low as 2 femtograms of DNA. Furthermore, ova from two different helminth genera were able to be detected as a multiplex assay using a single lateral flow strip, which could significantly reduce the time and the cost of helminth identification. The RPA-LF system represents an accurate, rapid, and cost-effective technology that could replace the existing detection methods, which are technically challenged and not ideal for on-site detection in wastewater treatment plants.

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A modified approach to recover and enumerate Ascaris ova in wastewater and sludge

Cited by 8 publications

References 16 publications

Real-time detection and identification of nematode eggs genus and species through optical imaging

Real-time detection and identification of nematode eggs genus and species through optical imaging

Detection of helminth ova genera using in-situ biosynthesis of gold nanoparticles

Detection of Helminth Ova in Wastewater Using Recombinase Polymerase Amplification Coupled to Lateral Flow Strips

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