Abstract
The objective of this study was to compare the results of an in vitro egg hatch test (EHT), micro-agar larval development test (MALDT) and in vivo faecal egg count reduction test (FECRT) between worm strains obtained from goats and sheep identically infected with the gastrointestinal parasitic nematode Haemonchus contortus. Results from the in vivo and in vitro tests were compared with benzimidazole (BZ)-resistance-associated β-tubulin allele frequencies determined using Pyrosequencing™. BZ resistance was not detected by the in vivo FECRT, where reductions of > 99% for both the resistant and the susceptible H. contortus strains were detected in both species. Discriminating doses in EHT and MALDT for the resistant strain indicated a low level (approx. 25%) of resistant individuals. Genotyping indicated that the susceptible strain had 10% BZ-resistant β-tubulin codon 200 alleles and the resistant strain had 26% respective resistant alleles. The in vitro tests and allele-frequency distribution suggested low levels of resistance in both strains; however, the FECRT did not support the evidence of resistant individuals of either strain in either species, suggesting a potential underestimation of low-level resistance in sheep and goats when employing this test.
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Acknowledgements
The authors thank S. Spišáková and M. Krčmárik for their technical assistance. The English has been revised throughout the whole manuscript by a native English language editor, Dr. William Blackhall.
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This study was supported by funds from the Scientific Grant Agency VEGA 2/0099/19 and Slovak Research and Development Agency APVV-18-0131.
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The Ethics Committee of the Institute of Parasitology of the Slovak Academy of Sciences approved animal use and experimental design under the European Community guidelines (EU Directive 2010/63/EU for animal experiments).
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Königová, A., Urda Dolinská, M., Babják, M. et al. Experimental evidence for the lack of sensitivity of in vivo faecal egg count reduction testing for the detection of early development of benzimidazole resistance. Parasitol Res 120, 153–159 (2021). https://doi.org/10.1007/s00436-020-06965-0
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DOI: https://doi.org/10.1007/s00436-020-06965-0