In vitro levamisole selection pressure on larval stages of Haemonchus contortus over nine generations gives rise to drug resistance and target site gene expression changes specific to the early larval stages only

Article


Sarai, Ranbir S., Kopp, Steven R., Knox, Malcolm R., Coleman, Glen T. and Kotze, Andrew C.. 2015. "In vitro levamisole selection pressure on larval stages of Haemonchus contortus over nine generations gives rise to drug resistance and target site gene expression changes specific to the early larval stages only." Veterinary Parasitology. 211 (1-2), pp. 45-53. https://doi.org/10.1016/j.vetpar.2015.05.002
Article Title

In vitro levamisole selection pressure on larval stages of Haemonchus contortus over nine generations gives rise to drug resistance and target site gene expression changes specific to the early larval stages only

ERA Journal ID5561
Article CategoryArticle
AuthorsSarai, Ranbir S. (Author), Kopp, Steven R. (Author), Knox, Malcolm R. (Author), Coleman, Glen T. (Author) and Kotze, Andrew C. (Author)
Journal TitleVeterinary Parasitology
Journal Citation211 (1-2), pp. 45-53
Number of Pages8
Year2015
Place of PublicationNetherlands
ISSN0304-4017
1873-2550
Digital Object Identifier (DOI)https://doi.org/10.1016/j.vetpar.2015.05.002
Abstract

There is some evidence that resistance to levamisole and pyrantel in trichostrongylid nematodes is due to changes in the composition of nicotinic acetylcholine receptors (nAChRs) which represent the drug target site. Altered expression patterns of genes coding for nAChR subunits, as well as the presence of truncated versions of several subunits, have been implicated in observed resistances. The studies have mostly compared target sites in worm isolates of very different genetic background, and hence the ability to associate the molecular changes with drug sensitivity alone have been clouded to some extent. The present study aimed to circumvent this issue by following target site gene expression pattern changes as resistance developed in Haemonchus contortus worms under laboratory selection pressure with levamisole. We applied drug selection pressure to early stage larvae in vitro over nine generations, and monitored changes in larval and adult drug sensitivities and target site gene expression patterns. High level resistance developed in larvae, with resistance factors of 94-fold and 1350-fold at the IC50 and IC95, respectively, in larval development assays after nine generations of selection. There was some cross-resistance to bephenium (70-fold increase in IC95). The expression of all the putative subunit components of levamisole-sensitive nAChRs, as well as a number of ancillary protein genes, particularly Hco-unc-29.1 and -ric-3, were significantly decreased (up to 5.5-fold) in the resistant larvae at generation nine compared to the starting population. However, adult worms did not show any resistance to levamisole, and showed an inverse pattern of gene expression changes, with many target site genes showing increased expression compared to the starting population. A comparison of the larval/adult drug sensitivity data with the known relationships for field-derived isolates indicated that the adults of our selected population should have been highly resistant to the drug if the larval/adult sensitivity relationships were in accordance with previous field isolates. Hence, our selected worms showed a life-stage drug sensitivity pattern quite different to that seen in the field. The present study has highlighted an association between drug target site changes and resistance to levamisole in H. contortus larvae. However, it has also highlighted the artificial nature of the larval selection method with levamisole, as the resistance phenotype and the associated molecular changes were only observed in the drug-pressured life stage. The study therefore reinforces the need for caution in extrapolating larval-based laboratory selection outcomes to field resistances.

Keywordsgene expression; haemonchus contortus; levamisole; nicotinic agonists; resistance; animals; antinematodal agents; drug resistance; gene expression regulation; haemonchiasis; haemonchus; helminth proteins; larva
ANZSRC Field of Research 2020300909. Veterinary parasitology
Public Notes

Files associated with this item cannot be displayed due to copyright restrictions.

Byline AffiliationsCommonwealth Scientific and Industrial Research Organisation (CSIRO), Australia
University of Queensland
Institution of OriginUniversity of Southern Queensland
Permalink -

https://research.usq.edu.au/item/q47w8/in-vitro-levamisole-selection-pressure-on-larval-stages-of-haemonchus-contortus-over-nine-generations-gives-rise-to-drug-resistance-and-target-site-gene-expression-changes-specific-to-the-early

  • 885
    total views
  • 10
    total downloads
  • 1
    views this month
  • 0
    downloads this month

Export as

Related outputs

Prevalence of canine heartworm infection in Queensland, Australia: comparison of diagnostic methods and investigation of factors associated with reduction in antigen detection
Constantinoiu, Constantin, Croton, Catriona, Paterson, Mandy B. A., Knott, Lyn, Henning, Joerg, Mallyon, John and Coleman, Glen T.. 2023. "Prevalence of canine heartworm infection in Queensland, Australia: comparison of diagnostic methods and investigation of factors associated with reduction in antigen detection." Parasites and Vectors. 16 (1). https://doi.org/10.1186/s13071-022-05633-9
Identification and prevalence of a gammaherpesvirus in free-ranging northern brown bandicoots (Isoodon macrourus)
Langhorne, Charlotte, Sullivan, Jennifer, Hoy, Julia, Kopp, Steven, Murray, Peter and McMichael, Lee. 2021. "Identification and prevalence of a gammaherpesvirus in free-ranging northern brown bandicoots (Isoodon macrourus)." Journal of Wildlife Disease. 57 (4), pp. 912-916. https://doi.org/10.7589/JWD-D-20-00166
Haematology and serum biochemistry results for anaesthetised northern brown bandicoots (Isoodon macrourus) in south east Queensland
Langhorne, Charlotte, McMichael, Lee, Hoy, Julia, Kopp, Stephen and Murray, Peter. 2021. "Haematology and serum biochemistry results for anaesthetised northern brown bandicoots (Isoodon macrourus) in south east Queensland." Australian Veterinary Journal. 99 (9), pp. 408-411. https://doi.org/10.1111/avj.13098
Clinical pathology of captive endangered mahogany gliders (Petaurus gracilis) in responses to environmental change
McMichael, Lee, Adam, Darlene, Tribe, Andrew, Bynon, Brian, Bradshaw, Lana, Hoy, Julia, Murray, Peter and Kopp, Steven. 2021. "Clinical pathology of captive endangered mahogany gliders (Petaurus gracilis) in responses to environmental change." Journal of Zoo and Wildlife Medicine. 52 (1), pp. 268-275. https://doi.org/10.1638/2019-0163
The 'Toolbox' of strategies for managing Haemonchus contortus in goats: What's in and what's out
Kearney, P. E., Murray, P. J., Hoy, J. M., Hohenhaus, M. and Kotze, A.. 2016. "The 'Toolbox' of strategies for managing Haemonchus contortus in goats: What's in and what's out." Veterinary Parasitology. 220, pp. 93-107. https://doi.org/10.1016/j.vetpar.2016.02.028
Client perspectives on desirable attributes and skills of veterinary technologists in Australia: considerations for curriculum design
Clarke, Patricia M., Al-Alawneh, John, Pitt, Rachael E., Schull, Daniel N. and Coleman, Glen T.. 2015. "Client perspectives on desirable attributes and skills of veterinary technologists in Australia: considerations for curriculum design." Journal of Veterinary Medical Education. 42 (3), pp. 217-231. https://doi.org/10.3138/jvme.0115-001R
Susceptibility of cat fleas (Siphonaptera: Pulicidae) to Fipronil and Imidacloprid using adult and larval bioassays
Rust, M. K., Vetter, R., Denholm, I., Blagburn, B., Williamson, M. S., Kopp, S., Coleman, G., Hostetler, J., Davis, W., Mencke, N., Rees, R., Foit, S. and Tetzner, K.. 2014. "Susceptibility of cat fleas (Siphonaptera: Pulicidae) to Fipronil and Imidacloprid using adult and larval bioassays." Journal of Medical Entomology. 51 (3), pp. 638-643. https://doi.org/10.1603/ME13240
A survey of macrocyclic lactone efficacy in Australian cyathostomin populations
Beasley, A. M., Kotze, A. C., Allen, K. and Coleman, G. T.. 2017. "A survey of macrocyclic lactone efficacy in Australian cyathostomin populations." Veterinary Parasitology: Regional Studies and Reports. 8, pp. 127-132. https://doi.org/10.1016/j.vprsr.2017.03.009
Adaptation of a 96-well plate larval migration inhibition test for measuring the sensitivity of cyathostomins to macrocyclic lactone anthelmintics
Beasley, A. M., Coleman, G. T. and Kotze, A. C.. 2017. "Adaptation of a 96-well plate larval migration inhibition test for measuring the sensitivity of cyathostomins to macrocyclic lactone anthelmintics." Veterinary Parasitology. 245, pp. 55-61. https://doi.org/10.1016/j.vetpar.2017.08.010
Drug-efflux and target-site gene expression patterns in Haemonchus contortus larvae able to survive increasing concentrations of levamisole in vitro
Sarai, Ranbir S., Kopp, Steven R., Coleman, Glen T. and Kotze, Andrew C.. 2014. "Drug-efflux and target-site gene expression patterns in Haemonchus contortus larvae able to survive increasing concentrations of levamisole in vitro." International Journal for Parasitology: Drugs and Drug Resistance. 4 (2), pp. 77-84. https://doi.org/10.1016/j.ijpddr.2014.02.001
Suspected ivermectin resistance in a south-east Queensland parascaris equorum population
Beasley, A., Coleman, G. and Kotze, A. C.. 2015. "Suspected ivermectin resistance in a south-east Queensland parascaris equorum population." Australian Veterinary Journal. 93 (9), pp. 305-307. https://doi.org/10.1111/avj.12352
Susceptibility of adult cat fleas (Siphonaptera: Pulicidae) to insecticides and status of insecticide resistance mutations at the Rdl and knockdown resistance loci
Rust, Michael K., Vetter, Richard, Denholm, Ian, Blagburn, Byron, Williamson, Martin S., Kopp, Steven, Coleman, Glen, Hostetler, Joe, Davis, Wendell, Mencke, Norbert, Rees, Robert, Foit, Sabrina, Bohm, Claudia and Tetzner, Kathrin. 2015. "Susceptibility of adult cat fleas (Siphonaptera: Pulicidae) to insecticides and status of insecticide resistance mutations at the Rdl and knockdown resistance loci." Parasitology Research. 114, pp. 7-18. https://doi.org/10.1007/s00436-015-4512-1
Use of big data in the surveillance of veterinary diseases: early detection of tick paralysis in companion animals
Guernier, Vanina, Milinovich, Gabriel J., Santos, Marcos Antonio Bezerra, Haworth, Mark, Coleman, Glen and Soares Magalhaes, Ricardo J.. 2016. "Use of big data in the surveillance of veterinary diseases: early detection of tick paralysis in companion animals." Parasites and Vectors. 9 (1). https://doi.org/10.1186/s13071-016-1590-6
Preventive health care of Pony Club horses in rural New South Wales, Australia
Buckley, P., Buckley, D., Coleman, G. T. and Morton, J. M.. 2016. "Preventive health care of Pony Club horses in rural New South Wales, Australia." Australian Veterinary Journal. 94 (8), pp. 265-270. https://doi.org/10.1111/avj.12464
Temporal variation in physiological biomarkers in black flying-foxes (Pteropus alecto), Australia
McMichael, Lee, Edson, Daniel, Mayer, David, McLaughlin, Amanda, Goldspink, Lauren, Vidgen, Miranda E., Kopp, Steven, Meers, Joanne and Field, Hume. 2016. "Temporal variation in physiological biomarkers in black flying-foxes (Pteropus alecto), Australia." Ecohealth. 13 (1), pp. 49-59. https://doi.org/10.1007/s10393-016-1113-0