Davis, CN and Tyson, F and Cutress, D and Davies, E and Jones, DL and Brophy, PM and Prescott, A and Rose, M and Williams, M and Williams, HW and Jones, RA, Rapid detection of Galba truncatula in water sources on pasture-land using loop-mediated isothermal amplification for control of trematode infections, Parasites and Vectors, 13, (1) Article 496. ISSN 1756-3305 (2020) [Refereed Article]
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Background: Fascioliasis caused by the trematodes Fasciola hepatica and F. gigantica, is a global neglected zoonotic disease estimated to cost the livestock industry over €2.5 billion annually. Farm management measures and sustainable use of anthelmintics can, in principle, effectively control trematode infection in livestock and reduce the rate of developing anthelmintic resistance. Previously, we designed an environmental DNA (eDNA) assay to identify a common trematode intermediate host, the freshwater snail Galba truncatula, in water sources to measure specific trematode infection risk areas on pasture-land. To improve this procedure, we now report a loop-mediated isothermal amplification (LAMP) assay to identify G. truncatula eDNA.
Methods: A LAMP assay was designed and optimised (e.g. temperature, time duration and primer concentration) to identify G. truncatula DNA. The ability of the LAMP assay to target G. truncatula DNA was identified, and LAMP assay limit of detection was investigated in comparison to conventional PCR. In the field, 48 water samples were collected from stream, ditch and water pool habitats in four locations at two Aberystwyth University farms over a seven week period to investigate the applicability of the LAMP assay for use on eDNA samples, in comparison to conventional PCR.
Results: The LAMP assay delivered detectable results in 30 min at 63 °C. The assay discriminated between G. truncatula DNA and non-target DNA, presenting a level of DNA detection comparable to conventional PCR. No significant difference was found between the ability of the LAMP and PCR assay to identify G. truncatula eDNA in water samples. Kappa coefficient analysis revealed a moderate level of agreement between LAMP and PCR assays.
Conclusions: This study demonstrated that the LAMP assay can detect G. truncatula eDNA in a simple and rapid manner. The LAMP assay may become a valuable tool to determine optimum pasture management for trematode parasite control.
|Item Type:||Refereed Article|
|Keywords:||loop-mediated isothermal amplification, environmental DNA, farm management, trematode, fluke, Galba truncatula, Fasciola hepatica, Calicophoron daubneyi|
|Research Division:||Agricultural, Veterinary and Food Sciences|
|Research Group:||Agricultural biotechnology|
|Research Field:||Agricultural biotechnology diagnostics (incl. biosensors)|
|Objective Division:||Animal Production and Animal Primary Products|
|Objective Group:||Livestock raising|
|Objective Field:||Sheep for meat|
|UTAS Author:||Rose, M (Dr Michael Rose)|
|Web of Science® Times Cited:||3|
|Deposited By:||Office of the Tasmanian Institute of Agriculture|
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