• 1.

    Amazigo U, 2008. The African Programme for Onchocerciasis Control (APOC). Ann Trop Med Parasitol 102 (Suppl 1 ):1922.

  • 2.

    Wolstenholme AJ, Rogers AT, 2005. Glutamate-gated chloride channels and the mode of action of the avermectin/milbemycin anthelmintics. Parasitology 131 (Suppl ):S85S95.

    • Search Google Scholar
    • Export Citation
  • 3.

    Shikiya K, Zaha O, Niimura S, Uehara T, Ohshiro J, Kinjo F, Saito A, Asato R, 1994. Clinical study on ivermectin against 125 strongyloidiasis patients. Kansenshogaku Zasshi 68: 1320.

    • Search Google Scholar
    • Export Citation
  • 4.

    Bourguinat C, Ardelli BF, Pion SD, Kamgno J, Gardon J, Duke BO, Boussinesq M, Prichard RK, 2008. P-glycoprotein-like protein, a possible genetic marker for ivermectin resistance selection in Onchocerca volvulus. Mol Biochem Parasitol 158: 101111.

    • Search Google Scholar
    • Export Citation
  • 5.

    Barnes EH, Dobson RJ, Stein PA, Le Jambre LF, Lenane IJ, 2001. Selection of different genotype larvae and adult worms for anthelmintic resistance by persistent and short-acting avermectin/milbemycins. Int J Parasitol 31: 720727.

    • Search Google Scholar
    • Export Citation
  • 6.

    Jember T, Nibret E, Amor A, Munshea A, Anegagrie M, 2020. Efficacy of single dose ivermectin against strongyloides stercoralis infection among primary school children in Amhara National Regional State. Infect Dis Res Treatment 13: 117863372093254.

    • Search Google Scholar
    • Export Citation
  • 7.

    Hays R, Esterman A, McDermott R, 2015. Type 2 diabetes mellitus is associated with Strongyloides stercoralis treatment failure in Australian aboriginals. PLoS Negl Trop Dis 9: e0003976.

    • Search Google Scholar
    • Export Citation
  • 8.

    Lespine A, Menez C, Bourguinat C, Prichard RK, 2012. P-glycoproteins and other multidrug resistance transporters in the pharmacology of anthelmintics: prospects for reversing transport-dependent anthelmintic resistance. Int J Parasitol Drugs Drug Resist 2: 5875.

    • Search Google Scholar
    • Export Citation
  • 9.

    Locher KP, 2016. Mechanistic diversity in ATP-binding cassette (ABC) transporters. Nat Struct Mol Biol 23: 487493.

  • 10.

    Palmeirim MS, Hurlimann E, Knopp S, Speich B, Belizario V Jr, Joseph SA, Vaillant M, Olliaro P, Keiser J, 2018. Efficacy and safety of co-administered ivermectin plus albendazole for treating soil-transmitted helminths: a systematic review, meta-analysis and individual patient data analysis. PLoS Negl Trop Dis 12: e0006458.

    • Search Google Scholar
    • Export Citation
  • 11.

    Reyes-Guerrero DE, Cedillo-Borda M, Alonso-Morales RA, Alonso-Diaz MA, Olmedo-Juarez A, Mendoza-de-Gives P, Lopez-Arellano ME, 2020. Comparative study of transcription profiles of the P-glycoprotein transporters of two Haemonchus contortus isolates: susceptible and resistant to ivermectin. Mol Biochem Parasitol 238: 111281.

    • Search Google Scholar
    • Export Citation
  • 12.

    Dicker AJ, Nath M, Yaga R, Nisbet AJ, Lainson FA, Gilleard JS, Skuce PJ, 2011. Teladorsagia circumcincta: the transcriptomic response of a multi-drug-resistant isolate to ivermectin exposure in vitro. Exp Parasitol 127: 351356.

    • Search Google Scholar
    • Export Citation
  • 13.

    Kerboeuf D, Guegnard F, 2011. Anthelmintics are substrates and activators of nematode P glycoprotein. Antimicrob Agents Chemother 55: 22242232.

    • Search Google Scholar
    • Export Citation
  • 14.

    Srivastava M, Misra-Bhattacharya S, 2015. Overcoming drug resistance for macro parasites. Future Microbiol 10: 17831789.

  • 15.

    James CE, Davey MW, 2009. Increased expression of ABC transport proteins is associated with ivermectin resistance in the model nematode Caenorhabditis elegans. Int J Parasitol 39: 213220.

    • Search Google Scholar
    • Export Citation
  • 16.

    Viney M, Kikuchi T, 2017. Strongyloides ratti and S. venezuelensis—rodent models of Strongyloides infection. Parasitology 144: 285294.

  • 17.

    Eamudomkarn C, Sithithaworn P, Sithithaworn J, Kaewkes S, Sripa B, Itoh M, 2015. Comparative evaluation of Strongyloides ratti and S. stercoralis larval antigen for diagnosis of strongyloidiasis in an endemic area of opisthorchiasis. Parasitol Res 114: 25432551.

    • Search Google Scholar
    • Export Citation
  • 18.

    Kaewrat W et al., 2020. Improved agar plate culture conditions for diagnosis of Strongyloides stercoralis. Acta Trop 203: 105291.

  • 19.

    Viney ME, Green LD, Brooks JA, Grant WN, 2002. Chemical mutagenesis of the parasitic nematode Strongyloides ratti to isolate ivermectin resistant mutants. Int J Parasitol 32: 16771682.

    • Search Google Scholar
    • Export Citation
  • 20.

    Schmittgen TD, Livak KJ, 2008. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3: 11011108.

  • 21.

    Pulaski CN et al., 2014. Establishment of macrocyclic lactone resistant Dirofilaria immitis isolates in experimentally infected laboratory dogs. Parasit Vectors 7: 494.

    • Search Google Scholar
    • Export Citation
  • 22.

    Gonzalez-Canga A, Fernandez-Martinez N, Sahagun-Prieto A, Diez-Liebana MJ, Sierra-Vega M, Garcia-Vieitez JJ, 2009. A review of the pharmacological interactions of ivermectin in several animal species. Curr Drug Metab 10: 359368.

    • Search Google Scholar
    • Export Citation
  • 23.

    Raza A, Kopp SR, Bagnall NH, Jabbar A, Kotze AC, 2016. Effects of in vitro exposure to ivermectin and levamisole on the expression patterns of ABC transporters in Haemonchus contortus larvae. Int J Parasitol Drugs Drug Resist 6: 103115.

    • Search Google Scholar
    • Export Citation
  • 24.

    Bartley DJ, McAllister H, Bartley Y, Dupuy J, Menez C, Alvinerie M, Jackson F, Lespine A, 2009. P-glycoprotein interfering agents potentiate ivermectin susceptibility in ivermectin sensitive and resistant isolates of Teladorsagia circumcincta and Haemonchus contortus. Parasitology 136: 10811088.

    • Search Google Scholar
    • Export Citation
  • 25.

    Markov GV, Baskaran P, Sommer RJ, 2015. The same or not the same: lineage-specific gene expansions and homology relationships in multigene families in nematodes. J Mol Evol 80: 1836.

    • Search Google Scholar
    • Export Citation
  • 26.

    Figueiredo LA, Reboucas TF, Ferreira SR, Rodrigues-Luiz GF, Miranda RC, Araujo RN, Fujiwara RT, 2018. Dominance of P-glycoprotein 12 in phenotypic resistance conversion against ivermectin in Caenorhabditis elegans. PLoS One 13: e0192995.

    • Search Google Scholar
    • Export Citation
  • 27.

    Gupta DK, Patra AT, Zhu L, Gupta AP, Bozdech Z, 2016. DNA damage regulation and its role in drug-related phenotypes in the malaria parasites. Sci Rep 6: 23603.

    • Search Google Scholar
    • Export Citation
  • 28.

    Gao F, Wang R, Liu M, 2014. Trichinella spiralis, potential model nematode for epigenetics and its implication in metazoan parasitism. Front Physiol 4: 410.

    • Search Google Scholar
    • Export Citation
  • 29.

    Dabin J, Fortuny A, Polo SE, 2016. Epigenome maintenance in response to DNA damage. Mol Cell 62: 712727.

  • 30.

    Hunt VL et al., 2016. The genomic basis of parasitism in the Strongyloides clade of nematodes. Nat Genet 48: 299307.

  • 31.

    Sanpool O, Intapan PM, Rodpai R, Laoraksawong P, Sadaow L, Tourtip S, Piratae S, Maleewong W, Thanchomnang T, 2019. Dogs are reservoir hosts for possible transmission of human strongyloidiasis in Thailand: molecular identification and genetic diversity of causative parasite species. J Helminthol 94: e110.

    • Search Google Scholar
    • Export Citation
  • 32.

    Anselmi M et al., 2015. Mass administration of ivermectin for the elimination of onchocerciasis significantly reduced and maintained low the prevalence of Strongyloides stercoralis in Esmeraldas, Ecuador. PLoS Negl Trop Dis 9: e0004150.

    • Search Google Scholar
    • Export Citation
  • 33.

    Buonfrate D et al., 2019. Multiple-dose versus single-dose ivermectin for Strongyloides stercoralis infection (Strong Treat 1 to 4): a multicentre, open-label, phase 3, randomised controlled superiority trial. Lancet Infect Dis 19: 11811190.

    • Search Google Scholar
    • Export Citation
 
 
 

 

 
 
 

 

 

 

 

 

 

Repeated Ivermectin Treatment Induces Ivermectin Resistance in Strongyloides ratti by Upregulating the Expression of ATP-Binding Cassette Transporter Genes

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  • 1 Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand;
  • | 2 Medical Technology Unit of Bangkatum Hospital, Phitsanulok, Thailand;
  • | 3 Chronic Kidney Disease Prevention in the Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand;
  • | 4 Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand;
  • | 5 Faculty of Medical Technology, Nakhonratchasima College, Nakhon Ratchasima, Thailand;
  • | 6 Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand;
  • | 7 Science Program in Biomedical Science, Khon Kaen University, Khon Kaen, Thailand;
  • | 8 Department of Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand;
  • | 9 Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand

ABSTRACT.

Ivermectin (IVM) is a widely used anthelmintic. However, with widespread use comes the risk of the emergence of IVM resistance, particularly in strongyloidiasis. Adenosine triphosphate (ATP)-binding cassette (ABC) transporter genes play an important role in the IVM-resistance mechanism. Here, we aimed to establish an animal experimental model of IVM resistance by frequent treatment of Strongyloides ratti with subtherapeutic doses of IVM, resistance being evaluated by the expression levels of ABC transporter genes. Rats infected with S. ratti were placed in experimental groups as follows: 1) untreated control (control); 2) treated with the mutagen ethyl methanesulfonate (EMS); 3) injected with 100 µg/kg body weight of IVM (IVM); 4) treated with a combination of EMS and IVM (IVM+EMS). Parasites were evaluated after four generations. Extent of IVM resistance was assessed using IVM sensitivity, larval development, and expression of ABC genes. By the F4 generation, S. ratti in the IVM group exhibited significantly higher levels of IVM resistance than did other groups according to in vitro drug-sensitivity tests and inhibition of larval development (IC50 = 36.60 ng/mL; 95% CI: 31.6, 42.01). Expression levels of ABC isoform genes (ABCA, ABCF, and ABCG) were statistically significantly higher in the IVM-resistant line compared with the susceptible line. In conclusion, IVM subtherapeutic doses induced IVM resistance in S. ratti by the F4 generation with corresponding upregulation of some ABC isoform genes. The study provides a model for inducing and assessing drug resistance in Strongyloides.

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Author Notes

Address correspondence to Somchai Pinlaor, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. E-mail: psomec@kku.ac.th

Financial support: This study was granted by Faculty of Medicine, Khon Kaen University, Thailand (Grant number IN62135), the project of CKDNET (Grant number CKDNET2559007), and the Postgraduate Scholarship to Mr. Chatchawan Sengthong, Faculty of Medicine, Khon Kaen University, Thailand.

Authors’ addresses: Chatchawan Sengthong, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand, Medical Technology Unit of Bangkatum Hospital, Phitsanulok, Thailand, and Chronic Kidney Disease Prevention in the Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand, E-mail: chachasengthong@gmail.com. Manachai Yingklang, Nuttanan Hongsrichan, Thewarach Laha, and Somchai Pinlaor, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand, and Chronic Kidney Disease Prevention in the Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand, E-mails: manatao@gmail.com, nuttho@kku.ac.th, thewa_la@kku.ac.th, and psomec@kku.ac.th. Kitti Intuyod, Chronic Kidney Disease Prevention in the Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand, and Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand, E-mail: kittin@kku.ac.th. Ornuma Haonon, Chronic Kidney Disease Prevention in the Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand, and Faculty of Medical Technology, Nakhonratchasima College, Nakhon Ratchasima, Thailand, E-mail: ornuma.h@nmc.ac.th. Porntip Pinlaor, Chronic Kidney Disease Prevention in The Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand, and Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand, E-mail: porawa@kku.ac.th. Chanakan Jantawong, Chronic Kidney Disease Prevention in the Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand, and Science Program in Biomedical Science, Khon Kaen University, Khon Kaen, Thailand, E-mail: chanakan19@hotmail.com. Sirirat Anutrakulchai, Chronic Kidney Disease Prevention in the Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand, and Department of Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand, E-mail: sirirt_a@kku.ac.th. Ubon Cha’on, Chronic Kidney Disease Prevention in the Northeast of Thailand (CKDNET), Faculty of Medicine, Khon Kaen University, Khon Kaen Province, Thailand, and Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand, E-mail: ubocha@kku.ac.th. Paiboon Sithithaworn, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand, E-mail: paibsit@gmail.com.

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