Absence of Polymorphisms in Codons 167, 198 and 200 of All Seven β-Tubulin Isotypes of Benzimidazole Susceptible and Resistant Parascaris spp. Specimens from Australia.
- Journal Article
Summary
The research article explores the absence of specific genetic changes in a certain protein (-tubulin) in both benzimidazole-resistant and susceptible species of the Parascaris genus from Australia. The findings suggest that mechanisms leading to resistance to the benzimidazole class of anti-parasitic drugs vary between ascarids (Parascaris) and nematode parasites, such as strongyle worms.
Details and Methodology
A couple of key processes were used in the research:
- Genome and transcriptome data from WormBase ParaSite, an online repository for parasitic worm draft genomes and transcriptomes, were utilised to compare the complete β-tubulin gene sequence amongst Parascaris species.
- Adult Parascaris specimens were collected from horses in Australia after treatment with benzimidazole (susceptible group) or those surviving the benzimidazole treatment and later collected after ivermectin treatment (resistant group).
- These specimens were then genotyped to pinpoint any potential changes at specific codon positions (167, 198 and 200) of the β-tubulin gene. These particular positions were chosen since, in the strongyle species of nematodes, mutations at these codons are linked to resistance to the benzimidazole class of drugs.
Findings
In both the data extracted from WormBase ParaSite in addition to the specimens physically collected, no polymorphisms in the identified codons were detected. Specifically:
- Six out of seven β-tubulin genes showed a benzimidazole-susceptible genotype, indicating no deviation from the typical genetic sequence.
- The exception was the testis-specific β-tubulin D genes, which all encoded tyrosine at codon 200, and this was witnessed across the board independent of the benzimidazole-susceptibility phenotype of the Parascaris species.
Conclusion
The absence of β-tubulin polymorphisms in the stated positions in both resistant and susceptible Parascaris spp. specimens suggests that resistance to benzimidazoles in ascarids is not correlated to these identified changes as seen in strongyles. Therefore, the study concludes that different mechanisms are in play leading to benzimidazole resistance in ascarid nematodes versus strongyle nematodes. This opens up new avenues for targeted research to understand drug resistance in different classes of parasites.
Cite This Article
Publication
Researcher Affiliations
- Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, 14163 Berlin, Germany.
- Department of Parasitology (Veterinary), Ankara University Graduate School of Health Sciences, 06230 Ankara, Turkey.
- Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, 14163 Berlin, Germany.
- Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, 14163 Berlin, Germany.
- School of Agricultural, Environmental and Veterinary Sciences, Charles Sturt University, Wagga Wagga, NSW 2678, Australia.
- School of Agricultural, Environmental and Veterinary Sciences, Charles Sturt University, Wagga Wagga, NSW 2678, Australia.
- Institute for Parasitology and Tropical Veterinary Medicine, Freie Universität Berlin, 14163 Berlin, Germany.
Conflict of Interest Statement
References
- Slocombe JO. Pathogenesis of helminths in equines.. Vet Parasitol 1985 Aug;18(2):139-53.
- Hautala K, Näreaho A, Kauppinen O, Nielsen MK, Sukura A, Rajala-Schultz PJ. Risk factors for equine intestinal parasite infections and reduced efficacy of pyrantel embonate against Parascaris sp.. Vet Parasitol 2019 Sep;273:52-59.
- Saeed MA, Beveridge I, Abbas G, Beasley A, Bauquier J, Wilkes E, Jacobson C, Hughes KJ, El-Hage C, O'Handley R, Hurley J, Cudmore L, Carrigan P, Walter L, Tennent-Brown B, Nielsen MK, Jabbar A. Systematic review of gastrointestinal nematodes of horses from Australia.. Parasit Vectors 2019 Apr 29;12(1):188.
- Misuno E, Clark CR, Anderson SL, Jenkins E, Wagner B, Dembek K, Petrie L. Characteristics of parasitic egg shedding over a 1-year period in foals and their dams in 2 farms in central Saskatchewan.. Can Vet J 2018 Mar;59(3):284-292.
- Nielsen MK, Wang J, Davis R, Bellaw JL, Lyons ET, Lear TL, Goday C. Parascaris univalens--a victim of large-scale misidentification?. Parasitol Res 2014 Dec;113(12):4485-90.
- Bullini L, Nascetti G, Carrè S, Rumore F, Biocca E. Ricerche cariologiche ed elettroforetiche su Parascaris univalens e Parascaris equorum. Acc. Naz. Lincei Rend. Cl. Sc. Fis. Mat. Nat. 1978;65:133–141.
- von Samson-Himmelstjerna G, Janssen IJI, Ramünke S, Goday C, Borges FA, Koudela B, Niedźwiedź A, Tomczuk K, Studzińska MB, Kornas S, Krücken J. Very low intraspecific sequence variation in selected nuclear and mitochondrial Parascaris univalens genes.. Infect Genet Evol 2021 Nov;95:105035.
- Pimpinelli S, Goday C. Unusual kinetochores and chromatin diminution in Parascaris.. Trends Genet 1989 Sep;5(9):310-5.
- Matthews JB. Anthelmintic resistance in equine nematodes.. Int J Parasitol Drugs Drug Resist 2014 Dec;4(3):310-5.
- Peregrine AS, Molento MB, Kaplan RM, Nielsen MK. Anthelmintic resistance in important parasites of horses: does it really matter?. Vet Parasitol 2014 Mar 17;201(1-2):1-8.
- von Samson-Himmelstjerna G. Anthelmintic resistance in equine parasites - detection, potential clinical relevance and implications for control.. Vet Parasitol 2012 Apr 19;185(1):2-8.
- Raza A, Qamar AG, Hayat K, Ashraf S, Williams AR. Anthelmintic resistance and novel control options in equine gastrointestinal nematodes.. Parasitology 2019 Apr;146(4):425-437.
- Craig TM, Diamond PL, Ferwerda NS, Thompson JA. Evidence of Ivermectin Resistance by Parascaris equorum on a Texas Horse Farm. J. Equine Vet. Sci. 2007;27:67–71.
- Alanazi AD, Mukbel RM, Alyousif MS, AlShehri ZS, Alanazi IO, Al-Mohammed HI. A field study on the anthelmintic resistance of Parascaris spp. in Arab foals in the Riyadh region, Saudi Arabia.. Vet Q 2017 Dec;37(1):200-205.
- Armstrong SK, Woodgate RG, Gough S, Heller J, Sangster NC, Hughes KJ. The efficacy of ivermectin, pyrantel and fenbendazole against Parascaris equorum infection in foals on farms in Australia.. Vet Parasitol 2014 Oct 15;205(3-4):575-80.
- Martin F, Halvarsson P, Delhomme N, Höglund J, Tydén E. Exploring the β-tubulin gene family in a benzimidazole-resistant Parascaris univalens population.. Int J Parasitol Drugs Drug Resist 2021 Dec;17:84-91.
- Kotze AC, Hunt PW, Skuce P, von Samson-Himmelstjerna G, Martin RJ, Sager H, Krücken J, Hodgkinson J, Lespine A, Jex AR, Gilleard JS, Beech RN, Wolstenholme AJ, Demeler J, Robertson AP, Charvet CL, Neveu C, Kaminsky R, Rufener L, Alberich M, Menez C, Prichard RK. Recent advances in candidate-gene and whole-genome approaches to the discovery of anthelmintic resistance markers and the description of drug/receptor interactions.. Int J Parasitol Drugs Drug Resist 2014 Dec;4(3):164-84.
- Ridley RK, Dryden MW, Gabbert NH, Schoning P. Epidemiology and control of helminth parasites in Greyhound breeding farms. Compend. Contin. Educ. Pract. Vet. 1994;16:585–599.
- Collins JB, Jordan B, Baldwin L, Hebron C, Paras K, Vidyashankar AN, Kaplan RM. Resistance to fenbendazole in Ascaridia dissimilis, an important nematode parasite of turkeys.. Poult Sci 2019 Nov 1;98(11):5412-5415.
- Krücken J, Fraundorfer K, Mugisha JC, Ramünke S, Sifft KC, Geus D, Habarugira F, Ndoli J, Sendegeya A, Mukampunga C, Bayingana C, Aebischer T, Demeler J, Gahutu JB, Mockenhaupt FP, von Samson-Himmelstjerna G. Reduced efficacy of albendazole against Ascaris lumbricoides in Rwandan schoolchildren.. Int J Parasitol Drugs Drug Resist 2017 Dec;7(3):262-271.
- Kwa MS, Veenstra JG, Van Dijk M, Roos MH. Beta-tubulin genes from the parasitic nematode Haemonchus contortus modulate drug resistance in Caenorhabditis elegans.. J Mol Biol 1995 Mar 3;246(4):500-10.
- Kwa MS, Veenstra JG, Roos MH. Benzimidazole resistance in Haemonchus contortus is correlated with a conserved mutation at amino acid 200 in beta-tubulin isotype 1.. Mol Biochem Parasitol 1994 Feb;63(2):299-303.
- Pape M, Posedi J, Failing K, Schnieder T, von Samson-Himmelstjerna G. Analysis of the beta-tubulin codon 200 genotype distribution in a benzimidazole-susceptible and -resistant cyathostome population.. Parasitology 2003 Jul;127(Pt 1):53-9.
- Prichard R. Genetic variability following selection of Haemonchus contortus with anthelmintics.. Trends Parasitol 2001 Sep;17(9):445-53.
- Ghisi M, Kaminsky R, Mäser P. Phenotyping and genotyping of Haemonchus contortus isolates reveals a new putative candidate mutation for benzimidazole resistance in nematodes.. Vet Parasitol 2007 Mar 31;144(3-4):313-20.
- Martínez-Valladares M, Valderas-García E, Gandasegui J, Skuce P, Morrison A, Castilla Gómez de Agüero V, Cambra-Pellejà M, Balaña-Fouce R, Rojo-Vázquez FA. Teladorsagia circumcincta beta tubulin: the presence of the E198L polymorphism on its own is associated with benzimidazole resistance.. Parasit Vectors 2020 Sep 7;13(1):453.
- Mohammedsalih KM, Krücken J, Bashar A, Juma FR, Abdalmalaik AAH, Khalafalla A, Abakar A, Coles G, von Samson-Himmelstjerna G. Susceptible trichostrongyloid species mask presence of benzimidazole-resistant Haemonchus contortus in cattle.. Parasit Vectors 2021 Feb 8;14(1):101.
- Mohammedsalih KM, Krücken J, Khalafalla A, Bashar A, Juma FR, Abakar A, Abdalmalaik AAH, Coles G, von Samson-Himmelstjerna G. New codon 198 β-tubulin polymorphisms in highly benzimidazole resistant Haemonchus contortus from goats in three different states in Sudan.. Parasit Vectors 2020 Mar 2;13(1):114.
- Avramenko RW, Redman EM, Melville L, Bartley Y, Wit J, Queiroz C, Bartley DJ, Gilleard JS. Deep amplicon sequencing as a powerful new tool to screen for sequence polymorphisms associated with anthelmintic resistance in parasitic nematode populations.. Int J Parasitol 2019 Jan;49(1):13-26.
- Redman E, Whitelaw F, Tait A, Burgess C, Bartley Y, Skuce PJ, Jackson F, Gilleard JS. The emergence of resistance to the benzimidazole anthlemintics in parasitic nematodes of livestock is characterised by multiple independent hard and soft selective sweeps.. PLoS Negl Trop Dis 2015 Feb;9(2):e0003494.
- Baltrušis P, Halvarsson P, Höglund J. Exploring benzimidazole resistance in Haemonchus contortus by next generation sequencing and droplet digital PCR.. Int J Parasitol Drugs Drug Resist 2018 Dec;8(3):411-419.
- Fávero FC, Dos Santos LB, Araújo FR, Ramünke S, Krücken J, von Samson-Himmelstjerna G, Borges FA. Haemonchus sp. in beef cattle in Brazil: species composition and frequency of benzimidazole resistance alleles.. Prev Vet Med 2020 Dec;185:105162.
- Ramünke S, Melville L, Rinaldi L, Hertzberg H, de Waal T, von Samson-Himmelstjerna G, Cringoli G, Mavrot F, Skuce P, Krücken J, Demeler J. Benzimidazole resistance survey for Haemonchus, Teladorsagia and Trichostrongylus in three European countries using pyrosequencing including the development of new assays for Trichostrongylus.. Int J Parasitol Drugs Drug Resist 2016 Dec;6(3):230-240.
- Esteban-Ballesteros M, Rojo-Vázquez FA, Skuce PJ, Melville L, González-Lanza C, Martínez-Valladares M. Quantification of resistant alleles in the β-tubulin gene of field strains of gastrointestinal nematodes and their relation with the faecal egg count reduction test.. BMC Vet Res 2017 Mar 20;13(1):71.
- Ademola IO, Krücken J, Ramünke S, Demeler J, von Samson-Himmelstjerna G. Absence of detectable benzimidazole-resistance associated alleles in Haemonchus placei in cattle in Nigeria revealed by pyrosequencing of β-tubulin isotype 1.. Parasitol Res 2015 May;114(5):1997-2001.
- Martínez-Valladares M, Donnan A, Geldhof P, Jackson F, Rojo-Vázquez FA, Skuce P. Pyrosequencing analysis of the beta-tubulin gene in Spanish Teladorsagia circumcincta field isolates.. Vet Parasitol 2012 Mar 23;184(2-4):371-6.
- Barrère V, Alvarez L, Suarez G, Ceballos L, Moreno L, Lanusse C, Prichard RK. Relationship between increased albendazole systemic exposure and changes in single nucleotide polymorphisms on the β-tubulin isotype 1 encoding gene in Haemonchus contortus.. Vet Parasitol 2012 May 25;186(3-4):344-9.
- Chaudhry U, Redman EM, Raman M, Gilleard JS. Genetic evidence for the spread of a benzimidazole resistance mutation across southern India from a single origin in the parasitic nematode Haemonchus contortus.. Int J Parasitol 2015 Sep;45(11):721-8.
- Chaudhry U, Miller M, Yazwinski T, Kaplan R, Gilleard J. The presence of benzimidazole resistance mutations in Haemonchus placei from US cattle.. Vet Parasitol 2014 Aug 29;204(3-4):411-5.
- Baltrušis P, Halvarsson P, Höglund J. Utilization of droplet digital PCR to survey resistance associated polymorphisms in the β tubulin gene of Haemonchus contortus in sheep flocks in Sweden.. Vet Parasitol 2020 Dec;288:109278.
- Sargison ND, MacLeay M, Morrison AA, Bartley DJ, Evans M, Chaudhry U. Development of amplicon sequencing for the analysis of benzimidazole resistance allele frequencies in field populations of gastrointestinal nematodes.. Int J Parasitol Drugs Drug Resist 2019 Aug;10:92-100.
- Diawara A, Halpenny CM, Churcher TS, Mwandawiro C, Kihara J, Kaplan RM, Streit TG, Idaghdour Y, Scott ME, Basáñez MG, Prichard RK. Association between response to albendazole treatment and β-tubulin genotype frequencies in soil-transmitted helminths.. PLoS Negl Trop Dis 2013;7(5):e2247.
- Roose S, Avramenko RW, Pollo SMJ, Wasmuth JD, Ame S, Ayana M, Betson M, Cools P, Dana D, Jones BP, Mekonnen Z, Morosetti A, Venkatesan A, Vlaminck J, Workentine ML, Levecke B, Gilleard JS, Geldhof P. Characterization of the β-tubulin gene family in Ascaris lumbricoides and Ascaris suum and its implication for the molecular detection of benzimidazole resistance.. PLoS Negl Trop Dis 2021 Sep;15(9):e0009777.
- Saunders GI, Wasmuth JD, Beech R, Laing R, Hunt M, Naghra H, Cotton JA, Berriman M, Britton C, Gilleard JS. Characterization and comparative analysis of the complete Haemonchus contortus β-tubulin gene family and implications for benzimidazole resistance in strongylid nematodes.. Int J Parasitol 2013 May;43(6):465-75.
- Driscoll M, Dean E, Reilly E, Bergholz E, Chalfie M. Genetic and molecular analysis of a Caenorhabditis elegans beta-tubulin that conveys benzimidazole sensitivity.. J Cell Biol 1989 Dec;109(6 Pt 1):2993-3003.
- Dilks CM, Hahnel SR, Sheng Q, Long L, McGrath PT, Andersen EC. Quantitative benzimidazole resistance and fitness effects of parasitic nematode beta-tubulin alleles.. Int J Parasitol Drugs Drug Resist 2020 Dec;14:28-36.
- Dilks CM, Koury EJ, Buchanan CM, Andersen EC. Newly identified parasitic nematode beta-tubulin alleles confer resistance to benzimidazoles.. Int J Parasitol Drugs Drug Resist 2021 Dec;17:168-175.
- Hahnel SR, Zdraljevic S, Rodriguez BC, Zhao Y, McGrath PT, Andersen EC. Extreme allelic heterogeneity at a Caenorhabditis elegans beta-tubulin locus explains natural resistance to benzimidazoles.. PLoS Pathog 2018 Oct;14(10):e1007226.
- Li L, Lü L, Nadler SA, Gibson DI, Zhang LP, Chen HX, Zhao WT, Guo YN. Molecular Phylogeny and Dating Reveal a Terrestrial Origin in the Early Carboniferous for Ascaridoid Nematodes.. Syst Biol 2018 Sep 1;67(5):888-900.
- Wilkes EJA, Cowling A, Woodgate RG, Hughes KJ. Inhomogeneity of the density of Parascaris spp. eggs in faeces of individual foals and the use of hypothesis testing for treatment decision making.. Vet Parasitol 2016 Oct 15;229:131-138.
- Wang J, Mitreva M, Berriman M, Thorne A, Magrini V, Koutsovoulos G, Kumar S, Blaxter ML, Davis RE. Silencing of germline-expressed genes by DNA elimination in somatic cells.. Dev Cell 2012 Nov 13;23(5):1072-80.
- Wang J, Gao S, Mostovoy Y, Kang Y, Zagoskin M, Sun Y, Zhang B, White LK, Easton A, Nutman TB, Kwok PY, Hu S, Nielsen MK, Davis RE. Comparative genome analysis of programmed DNA elimination in nematodes.. Genome Res 2017 Dec;27(12):2001-2014.
- Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool.. J Mol Biol 1990 Oct 5;215(3):403-10.
- Howe KL, Bolt BJ, Shafie M, Kersey P, Berriman M. WormBase ParaSite - a comprehensive resource for helminth genomics.. Mol Biochem Parasitol 2017 Jul;215:2-10.
- Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets.. Mol Biol Evol 2016 Jul;33(7):1870-4.
- Xia X. DAMBE7: New and Improved Tools for Data Analysis in Molecular Biology and Evolution.. Mol Biol Evol 2018 Jun 1;35(6):1550-1552.
- Xia X, Lemey P. Assessing substitution saturation with DAMBE. In: Lemey P., Salemi M., Vandamme A.-M., editors. The Phylogenetic Handbook: A Practical Approach to Phylogenetic Analysis and Hypothesis Testing. Cambridge University Press; Cambridge, UK: 2009. pp. 615–630.
- Xia X, Xie Z, Salemi M, Chen L, Wang Y. An index of substitution saturation and its application.. Mol Phylogenet Evol 2003 Jan;26(1):1-7.
- Castresana J. Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis.. Mol Biol Evol 2000 Apr;17(4):540-52.
- Nguyen LT, Schmidt HA, von Haeseler A, Minh BQ. IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies.. Mol Biol Evol 2015 Jan;32(1):268-74.
- Trifinopoulos J, Nguyen LT, von Haeseler A, Minh BQ. W-IQ-TREE: a fast online phylogenetic tool for maximum likelihood analysis.. Nucleic Acids Res 2016 Jul 8;44(W1):W232-5.
- Kalyaanamoorthy S, Minh BQ, Wong TKF, von Haeseler A, Jermiin LS. ModelFinder: fast model selection for accurate phylogenetic estimates.. Nat Methods 2017 Jun;14(6):587-589.
- Hoang DT, Chernomor O, von Haeseler A, Minh BQ, Vinh LS. UFBoot2: Improving the Ultrafast Bootstrap Approximation.. Mol Biol Evol 2018 Feb 1;35(2):518-522.
- Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, Gascuel O. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0.. Syst Biol 2010 May;59(3):307-21.
- Anisimova M, Gil M, Dufayard JF, Dessimoz C, Gascuel O. Survey of branch support methods demonstrates accuracy, power, and robustness of fast likelihood-based approximation schemes.. Syst Biol 2011 Oct;60(5):685-99.
- Beech RN, Wolstenholme AJ, Neveu C, Dent JA. Nematode parasite genes: what's in a name?. Trends Parasitol 2010 Jul;26(7):334-40.
- Vercruysse J, Albonico M, Behnke JM, Kotze AC, Prichard RK, McCarthy JS, Montresor A, Levecke B. Is anthelmintic resistance a concern for the control of human soil-transmitted helminths?. Int J Parasitol Drugs Drug Resist 2011 Dec;1(1):14-27.
- . World Health Organisation Guideline: Preventive Chemotherapy to Control Soil-Transmitted Helminth Infections in At-Risk Population Groups. [(accessed on 20 March 2022)]. Available online: https://apps.who.int/iris/handle/10665/258983.
- Ma Z, Michailides TJ. Advances in understanding molecular mechanisms of fungicide resistance and molecular detection of resistant genotypes in phytopathogenic fungi. Crop Prot. 2005;24:853–863.
- Stasiuk SJ, MacNevin G, Workentine ML, Gray D, Redman E, Bartley D, Morrison A, Sharma N, Colwell D, Ro DK, Gilleard JS. Similarities and differences in the biotransformation and transcriptomic responses of Caenorhabditis elegans and Haemonchus contortus to five different benzimidazole drugs.. Int J Parasitol Drugs Drug Resist 2019 Dec;11:13-29.
- Stuchlíková LR, Matoušková P, Vokřál I, Lamka J, Szotáková B, Sečkařová A, Dimunová D, Nguyen LT, Várady M, Skálová L. Metabolism of albendazole, ricobendazole and flubendazole in Haemonchus contortus adults: Sex differences, resistance-related differences and the identification of new metabolites.. Int J Parasitol Drugs Drug Resist 2018 Apr;8(1):50-58.
- Kellerová P, Raisová Stuchlíková L, Matoušková P, Štěrbová K, Lamka J, Navrátilová M, Vokřál I, Szotáková B, Skálová L. Sub-lethal doses of albendazole induce drug metabolizing enzymes and increase albendazole deactivation in Haemonchus contortus adults.. Vet Res 2020 Jul 23;51(1):94.
Citations
This article has been cited 1 times.- Cain JL, Nielsen MK. The equine ascarids: resuscitating historic model organisms for modern purposes.. Parasitol Res 2022 Oct;121(10):2775-2791.