Abstract
The sheep body louse, Bovicola ovis (B. ovis), is one of the most significant ectoparasites affecting Australia’s sheep flocks. Despite this, detection methods for B. ovis infestation are limited to visual inspection and ELISA. A colourimetric loop-mediated isothermal amplification (LAMP) method was developed and evaluated for the detection of B. ovis DNA. Diagnostic sensitivity and specificity of LAMP were compared with those of visual inspection and PCR and validated using field samples collected from 22 farms. Two different DNA extraction methods using a commercial kit and a boiling method were also compared. The highest sensitivity and specificity were observed when PCR was used and DNA was extracted using a commercial kit. Compared with PCR, the LAMP assay demonstrated a sensitivity and specificity of 90% and 92% when DNA was extracted by a commercial kit and 100% and 75% when DNA was extracted by the boiling method, respectively. The LAMP test developed in this study could potentially serve as a point-of-care diagnostic tool for monitoring of sheep flocks as well as surveillance of B. ovis populations.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abdelhai M, Hassanin HM, Sun X (2016) Comparative study of rapid DNA extraction methods of pathogenic bacteria. Am J Biosci Bioeng 4:1–8
Beath K (2017) randomLCA : an R package for latent class with random effects analysis. J Stat Softw 81. https://doi.org/10.18637/jss.v081.i13
Chaouch M, Aoun K, Ben Othman S, Ben Abid M, Ben Sghaier I, Bouratbine A, Ben Abderrazak S (2019) Development and assessment of Leishmania major- and Leishmania tropica-specific loop-mediated isothermal amplification assays for the diagnosis of cutaneous leishmaniasis in Tunisia. Am J Trop Med Hyg. https://doi.org/10.4269/ajtmh.19-0097
Cotter J (2019) Sheep lice, Biosecurity can prevent introduction. http://www.liceboss.com.au/sheep-goats/prevention/sheep-lice-biosecurity-can-prevent-introduction.php. Accessed Aug 2019
DAWR (2018) Wool (FACT Sheet) Department of Agriculture and Water Resources, Australian Government. http://www.agriculture.gov.au/about/commitment/portfolio-facts/wool. Accessed Aug 2019
James PJ (2013) Biology of sheep lice (Bovicola ovis). LiceBoss. http://www.liceboss.com.au/files/pages/notes/Biology_of_sheep_lice_Bovicola_ovis.pdf. Accessed Aug 2019
James PJ, Garrett JA, Moon RD (2002) Sensitivity of two-stage sampling to detect sheep biting lice (Bovicola ovis) in infested flocks. Vet Parasitol 103:157–166. https://doi.org/10.1016/S0304-4017(01)00585-4
Joshua E, Junk G, Levot G (2010) Sheep lice (primefacts). Department of Primary Industries, NSW. https://www.dpi.nsw.gov.au/__data/assets/pdf_file/0005/318704/Sheep-lice.pdf. Accessed Aug 2019
Lane J, Jubb T, Shephard R, Webb-Ware J, Fordyce G (2015) Priority list of endemic diseases for the red meat industries. Meat and Livestock Australia Ltd, Sydney
Martzy R et al (2017) A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Enterococcus spp. in water. Water Res 122:62–69. https://doi.org/10.1016/j.watres.2017.05.023
Morcombe P, Young G, Ball M, Dunlop R (1996) The detection of lice (Bovicola ovis) in mobs of sheep: a comparison of fleece parting, the lamp test and the table locks test. Aust Vet J 73:170–173. https://doi.org/10.1111/j.1751-0813.1996.tb10020.x
Niessen L, Luo J, Denschlag C, Vogel RF (2013) The application of loop-mediated isothermal amplification (LAMP) in food testing for bacterial pathogens and fungal contaminants. Food Microbiol 36:191–206. https://doi.org/10.1016/j.fm.2013.04.017
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:e63–e63. https://doi.org/10.1093/nar/28.12.e63
Popp S, Eppleston J, Watt BR, Mansfield S, Bush RD (2012) The prevalence of lice (Bovicola ovis) in sheep flocks on the central and southern Tablelands of New South Wales. Ani Prod Sci 52:659–664. https://doi.org/10.1071/AN11240
Pritt BS (2015) Chapter 4 - Molecular diagnostics in the diagnosis of parasitic infection. In: Sails A, Tang Y-W (eds) Methods in Microbiology, vol 42. Academic Press, pp 111–160. https://doi.org/10.1016/bs.mim.2015.05.001
Reeve I, Walkden-Brown, S. (2014) Benchmarking Australian sheep parasite control: cross-sectional survey report Meat and Livestock Australia Limited Sydney
Sepahvand A, Pestehchian N, Yousefi HA, Gharehbaba RP (2017) Comparison and evaluation of four methods for extracting DNA from Giardia duodenalis cysts for PCR targeting the tpi gene. J Parasit Dis 41:263–267. https://doi.org/10.1007/s12639-016-0790-5
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680. https://doi.org/10.1093/nar/22.22.4673
Wojtek PM, Paul Y, Shiell B, Levot G (2001) Development of a lice detection test for ‘on-farm’. Paper presented at the FLICS Conference Launceston,
Zhu K, Jin H, He Z, Zhu Q, Wang B (2006) A continuous method for the large-scale extraction of plasmid DNA by modified boiling lysis. Nat Protoc 1:3088–3093. https://doi.org/10.1038/nprot.2006.452
Acknowledgements
The authors would like to thank Ag resellers and the farmers from the Riverina in NSW, who participated in this study by providing samples.
Funding
This work was supported by the Graham Centre for Agricultural Innovation (grant number 50714) and the School of Animal and Veterinary Sciences (grant number 40702) at Charles Sturt University.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
All the experiments complied with the current policies of Charles Sturt University.
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Section Editor: Douglas D. Colwell
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Wong, S.A., Woodgate, R.G., Pant, S.D. et al. Rapid detection of Bovicola ovis using colourimetric loop-mediated isothermal amplification (LAMP): a potential tool for the detection of sheep lice infestation on farm. Parasitol Res 119, 395–401 (2020). https://doi.org/10.1007/s00436-019-06552-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-019-06552-y