title: Rapid concentration and sensitive detection of hookworm ova from wastewater matrices using a real-time PCR method.
abstract: The risk of human hookworm infections from land application of wastewater matrices could be high in regions with high hookworm prevalence. A rapid, sensitive and specific hookworm detection method from wastewater matrices is required in order to assess human health risks. Currently available methods used to identify hookworm ova to the species level are time consuming and lack accuracy. In this study, a real-time PCR method was developed for the rapid, sensitive and specific detection of canine hookworm (Ancylostoma caninum) ova from wastewater matrices. A. caninum was chosen because of its morphological similarity to the human hookworm (Ancylostoma duodenale and Necator americanus). The newly developed PCR method has high detection sensitivity with the ability to detect less than one A. caninum ova from 1 L of secondary treated wastewater at the mean threshold cycle (CT) values ranging from 30.1 to 34.3. The method is also able to detect four A. caninum ova from 1 L of raw wastewater and from ∼4 g of treated sludge with mean CT values ranging from 35.6 to 39.8 and 39.8 to 39.9, respectively. The better detection sensitivity obtained for secondary treated wastewater compared to raw wastewater and sludge samples could be attributed to sample turbidity. The proposed method appears to be rapid, sensitive and specific compared to traditional methods and has potential to aid in the public health risk assessment associated with land application of wastewater matrices. Furthermore, the method can be adapted to detect other helminth ova of interest from wastewater matrices.
authors: CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia; School of Public Health, The University of Queensland, Herston Road, QLD 4006, Australia. Electronic address: Pradip.Gyawali@csiro.au.
authors: P
authors: P
authors: Gyawali
authors: CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia; School of Public Health, The University of Queensland, Herston Road, QLD 4006, Australia.
authors: J P S
authors: JP
authors: Sidhu
authors: CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia; Faculty of Science, Health and Education, University of the Sunshine Coast, Maroochydore, DC, QLD 4558, Australia.
authors: W
authors: W
authors: Ahmed
authors: School of Public Health, The University of Queensland, Herston Road, QLD 4006, Australia.
authors: P
authors: P
authors: Jagals
authors: CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia; School of Public Health, The University of Queensland, Herston Road, QLD 4006, Australia.
authors: S
authors: S
authors: Toze
mesh_terms: D000722:Ancylostoma / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D018509:DNA, Helminth / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000302:isolation & purification; D004285:Dogs; D005243:Feces / Q000469:parasitology; D057230:Limit of Detection; D010063:Ovum; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012722:Sewage / Q000469:parasitology; D062065:Wastewater / Q000469:parasitology*
chemical_list: D018509:DNA, Helminth; D004275:DNA, Ribosomal; D012340:RNA, Ribosomal, 5.8S; D012722:Sewage; D062065:Wastewater
keywords: Health risk assessment; Helminth; Hookworm; Real-time PCR; Sludge; Wastewater