• 1.

    Pullan RL, Smith JL, Jasrasaria R, Brooker SJ, 2014. Global numbers of infection and disease burden of soil transmitted helminth infections in 2010. Parasit Vectors 7: 37.

    • Search Google Scholar
    • Export Citation
  • 2.

    Brooker S, 2010. Estimating the global distribution and disease burden of intestinal nematode infections: adding up the numbers—a review. Int J Parasitol 40: 11371144.

    • Search Google Scholar
    • Export Citation
  • 3.

    Liu C et al. 2015. Soil-transmitted helminths in southwestern China: a cross-sectional study of links to cognitive ability, nutrition, and school performance among children. PLoS Negl Trop Dis 9: e0003877.

    • Search Google Scholar
    • Export Citation
  • 4.

    Campbell SJ, Nery SV, Doi SA, Gray DJ, Soares Magalhães RJ, McCarthy JS, Traub RJ, Andrews RM, Clements AC, 2016. Complexities and perplexities: a critical appraisal of the evidence for soil-transmitted helminth infection-related morbidity. PLoS Negl Trop Dis 10: e0004566.

    • Search Google Scholar
    • Export Citation
  • 5.

    World Health Organization, 2012. Soil-Transmitted Helminthiases: Eliminating Soil-Transmitted Helminthiases as a Public Health Problem in Children: Progress Report 2001–2010 and Strategic Plan 2011–2020. Geneva, Switzerland: WHO.

    • Search Google Scholar
    • Export Citation
  • 6.

    Taylor-Robinson DC, Maayan N, Soares-Weiser K, Donegan S, Garner P, 2015. Deworming drugs for soil-transmitted intestinal worms in children: effects on nutritional indicators, haemoglobin, and school performance. Cochrane Database Syst Rev 7: CD000371.

    • Search Google Scholar
    • Export Citation
  • 7.

    Campbell SJ, Nery SV, McCarthy JS, Gray DJ, Soares Magalhães RJ, Clements AC, 2016. A critical appraisal of control strategies for soil-transmitted helminths. Trends Parasitol 32: 97107.

    • Search Google Scholar
    • Export Citation
  • 8.

    Albonico M, Levecke B, LoVerde PT, Montresor A, Prichard R, Vercruysse J, Webster JP, 2015. Monitoring the efficacy of drugs for neglected tropical diseases controlled by preventive chemotherapy. J Glob Antimicrob Resist 3: 229236.

    • Search Google Scholar
    • Export Citation
  • 9.

    Vercruysse J, Albonico M, Behnke JM, Kotze AC, Prichard RK, McCarthy JS, Montresor A, Levecke B, 2011. Is anthelmintic resistance a concern for the control of human soil-transmitted helminths? Int J Parasitol Drugs Drug Resist 1: 1427.

    • Search Google Scholar
    • Export Citation
  • 10.

    Wolstenholme AJ, Martin RJ, 2014. Anthelmintics—from discovery to resistance. Int J Parasitol Drugs Drug Resist 4: 218219.

  • 11.

    Leathwick DM, Luo D, 2017. Managing anthelmintic resistance—variability in the dose of drug reaching the target worms influences selection for resistance? Vet Parasitol 243: 2935.

    • Search Google Scholar
    • Export Citation
  • 12.

    Rose H et al. 2015. Widespread anthelmintic resistance in European farmed ruminants: a systematic review. Vet Rec 176: 546.

  • 13.

    Knapp-Lawitzke F, Krucken J, Ramunke S, von Samson-Himmelstjerna G, Demeler J, 2015. Rapid selection for β-tubulin alleles in codon 200 conferring benzimidazole resistance in an Ostertagia ostertagi isolate on pasture. Vet Parasitol 209: 8492.

    • Search Google Scholar
    • Export Citation
  • 14.

    Barrere V, Alvarez L, Suarez G, Ceballos L, Moreno L, Lanusse C, Prichard RK, 2012. 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 186: 344349.

    • Search Google Scholar
    • Export Citation
  • 15.

    Robinson MW, McFerran N, Trudgett A, Hoey L, Fairweather I, 2004. A possible model of benzimidazole binding to beta-tubulin disclosed by invoking an inter-domain movement. J Mol Graph Model 23: 275284.

    • Search Google Scholar
    • Export Citation
  • 16.

    Lubega GW, Prichard RK, 1990. Specific interaction of benz-imidazole anthelmintics with tubulin: high-affinity binding and benzimidazole resistance in H. contortus. Mol Biochem Parasitol 38: 221232.

    • Search Google Scholar
    • Export Citation
  • 17.

    Schwab AE, Boakye DA, Kyelem D, Prichard RK, 2005. Detection of benzimidazole resistance-associated mutations in the filarial nematode Wuchereria bancrofti and evidence for selection by albendazole and ivermectin combination treatment. Am J Trop Med Hyg 73: 234238.

    • Search Google Scholar
    • Export Citation
  • 18.

    Nana-Djeunga H, Bourguinat C, Pion SD, Kamgno J, Gardon J, Njiokou F, Boussinesq M, Prichard RK, 2012. Single nucleotide polymorphisms in β-tubulin selected in Onchocerca volvulus following repeated ivermectin treatment: possible indication of resistance selection. Mol Biochem Parasitol 185: 1018.

    • Search Google Scholar
    • Export Citation
  • 19.

    Albonico M, Wright V, Bickle Q, 2004. Molecular analysis of the beta-tubulin gene of human hookworms as a basis for possible benzimidazole resistance on Pemba Island. Mol Biochem Parasitol 134: 281284.

    • Search Google Scholar
    • Export Citation
  • 20.

    Diawara A et al. 2013. Association between response to albendazole treatment and β-tubulin genotype frequencies in soil-transmitted helminths. PLoS Negl Trop Dis 7: e2247.

    • Search Google Scholar
    • Export Citation
  • 21.

    Diawara A, Schwenkenbecher JM, Kaplan RM, Prichard RK, 2013. Molecular and biological diagnostic tests for monitoring benzimidazole resistance in human soil-transmitted helminths. Am J Trop Med Hyg 88: 10521061.

    • Search Google Scholar
    • Export Citation
  • 22.

    Schwenkenbecher JM, Albonico M, Bickle Q, Kaplan RM, 2007. Characterization of beta-tubulin genes in hookworms and investigation of resistance-associated mutations using real-time PCR. Mol Biochem Parasitol 156: 167174.

    • Search Google Scholar
    • Export Citation
  • 23.

    Humphries D et al. 2013. Hookworm infection among school age children in Kintampo north municipality, Ghana: nutritional risk factors and response to albendazole treatment. Am J Trop Med Hyg 89: 540548.

    • Search Google Scholar
    • Export Citation
  • 24.

    Humphries D et al. 2011. Epidemiology of hookworm infection in Kintampo north municipality, Ghana: patterns of malaria coinfection, anemia, and albendazole treatment failure. Am J Trop Med Hyg 84: 792800.

    • Search Google Scholar
    • Export Citation
  • 25.

    Humphries D, Nguyen S, Kumar S, Quagraine J, Otchere J, Harrison L, Wilson M, Cappello M, 2017. Effectiveness of albendazole against hookworm varies widely by community and correlates with nutritional factors: a cross-sectional study of school-age children in Ghana. Am J Trop Med Hyg 96: 347354.

    • Search Google Scholar
    • Export Citation
  • 26.

    World Health Organization, 2013. Assessing the Efficacy of Anthelminthic Drugs against Schistosomiasis and Soil-Transmitted Helminthiases. Geneva, Switzerland: WHO.

    • Search Google Scholar
    • Export Citation
  • 27.

    Chu D, Bungiro RD, Ibanez M, Harrison LM, Campodonico E, Jones BF, Mieszczanek J, Kuzmic P, Cappello M, 2004. Molecular characterization of Ancylostoma ceylanicum Kunitz-type serine protease inhibitor: evidence for a role in hookworm-associated growth delay. Infect Immun 72: 22142221.

    • Search Google Scholar
    • Export Citation
  • 28.

    Germer S, Holland MJ, Higuchi R, 2000. High-throughput SNP allele-frequency determination in pooled DNA samples by kinetic PCR. Genome Res 10: 258266.

    • Search Google Scholar
    • Export Citation
  • 29.

    Alvarez-Sanchez MA, Perez-Garcia J, Cruz-Rojo MA, Rojo-Vazquez FA, 2005. Real time PCR for the diagnosis of benzimidazole resistance in trichostrongylids of sheep. Vet Parasitol 129: 291298.

    • Search Google Scholar
    • Export Citation
  • 30.

    Roy S, Schreiber E, 2014. Detecting and quantifying low level gene variants in sanger sequencing traces using the ab1 peak reporter tool. J Biomol Tech 25 (Suppl 1): S13S14.

    • Search Google Scholar
    • Export Citation
  • 31.

    Kotze AC et al. 2014. 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 4: 164184.

    • Search Google Scholar
    • Export Citation
  • 32.

    Awadzi K et al. 2004. An investigation of persistent microfilaridermias despite multiple treatments with ivermectin, in two onchocerciasis-endemic foci in Ghana. Ann Trop Med Parasitol 98: 231249.

    • Search Google Scholar
    • Export Citation
  • 33.

    Rashwan N, Bourguinat C, Keller K, Gunawardena NK, de Silva N, Prichard R, 2016. Isothermal diagnostic assays for monitoring single nucleotide polymorphisms in Necator americanus associated with benzimidazole drug resistance. PLoS Negl Trop Dis 10: e0005113.

    • Search Google Scholar
    • Export Citation
  • 34.

    Kotze AC, Cowling K, Bagnall NH, Hines BM, Ruffell AP, Hunt PW, Coleman GT, 2012. Relative level of thiabendazole resistance associated with the E198A and F200Y SNPs in larvae of a multi-drug resistant isolate of Haemonchus contortus. Int J Parasitol Drugs Drug Resist 2: 9297.

    • Search Google Scholar
    • Export Citation
  • 35.

    Medley GF, Hollingsworth TD, 2015. MDA helminth control: more questions than answers. Lancet Glob Health 3: e583e584.

  • 36.

    Allen T, Parker M, 2016. Deworming delusions? Mass drug administration in east African schools. J Biosoc Sci 48 (Suppl 1): S116S147.

  • 37.

    Albonico M, Engels D, Savioli L, 2004. Monitoring drug efficacy and early detection of drug resistance in human soil-transmitted nematodes: a pressing public health agenda for helminth control. Int J Parasitol 34: 12051210.

    • Search Google Scholar
    • Export Citation
  • 38.

    Keiser J, Utzinger J, 2008. Efficacy of current drugs against soil-transmitted helminth infections: systematic review and meta-analysis. JAMA 299: 19371948.

    • Search Google Scholar
    • Export Citation
  • 39.

    Okoyo C et al. 2016. Monitoring the impact of a national school based deworming programme on soil-transmitted helminths in Kenya: the first three years, 2012–2014. Parasit Vectors 9: 113.

    • Search Google Scholar
    • Export Citation
  • 40.

    Truscott JE, Hollingsworth TD, Brooker SJ, Anderson RM, 2014. Can chemotherapy alone eliminate the transmission of soil transmitted helminths? Parasit Vectors 7: 266.

    • Search Google Scholar
    • Export Citation
  • 41.

    Quinnell RJ, Slater AF, Tighe P, Walsh EA, Keymer AE, Pritchard DI, 1993. Reinfection with hookworm after chemotherapy in Papua New Guinea. Parasitology 106: 379385.

    • Search Google Scholar
    • Export Citation
  • 42.

    Jia TW, Melville S, Utzinger J, King CH, Zhou XN, 2012. Soil-transmitted helminth reinfection after drug treatment: a systematic review and meta-analysis. PLoS Negl Trop Dis 6: e1621.

    • Search Google Scholar
    • Export Citation
  • 43.

    Drogemuller M, Failing K, Schnieder T, von Samson-Himmelstjerna G, 2004. Effect of repeated benzimidazole treatments with increasing dosages on the phenotype of resistance and the beta-tubulin codon 200 genotype distribution in a benzimidazole-resistant cyathostomin population. Vet Parasitol 123: 201213.

    • Search Google Scholar
    • Export Citation
  • 44.

    de Lourdes Mottier M, Prichard RK, 2008. Genetic analysis of a relationship between macrocyclic lactone and benzimidazole anthelmintic selection on Haemonchus contortus. Pharmacogenet Genomics 18: 129140.

    • Search Google Scholar
    • Export Citation
 
 
 

 

 
 
 

 

 

 

 

 

 

Genetic Markers of Benzimidazole Resistance among Human Hookworms (Necator americanus) in Kintampo North Municipality, Ghana

View More View Less
  • 1 Yale Partnerships for Global Health, Department of Pediatrics, Yale School of Medicine, New Haven, Connecticut;
  • | 2 Department of Parasitology, Noguchi Memorial Institute for Medical Research, University of Ghana, Legon, Ghana;
  • | 3 Department of Ecology and Evolutionary Biology, Yale University, New Haven, Connecticut;
  • | 4 Department of Epidemiology of Microbial Diseases, Yale School of Public Health, New Haven, Connecticut

Hookworm infection causes anemia, malnutrition, and growth delay, especially in children living in sub-Saharan Africa. The World Health Organization recommends periodic mass drug administration (MDA) of anthelminthics to school-age children (SAC) as a means of reducing morbidity. Recently, questions have been raised about the effectiveness of MDA as a global control strategy for hookworms and other soil-transmitted helminths (STHs). Genomic DNA was extracted from Necator americanus hookworm eggs isolated from SAC enrolled in a cross-sectional study of STH epidemiology and deworming response in Kintampo North Municipality, Ghana. A polymerase chain reaction (PCR) assay was then used to identify single-nucleotide polymorphisms (SNPs) associated with benzimidazole resistance within the N. americanus β-tubulin gene. Both F167Y and F200Y resistance–associated SNPs were detected in hookworm samples from infected study subjects. Furthermore, the ratios of resistant to wild-type SNP at these two loci were increased in posttreatment samples from subjects who were not cured by albendazole, suggesting that deworming drug exposure may enrich resistance-associated mutations. A previously unreported association between F200Y and a third resistance-associated SNP, E198A, was identified by sequencing of F200Y amplicons. These data confirm that markers of benzimidazole resistance are circulating among hookworms in central Ghana, with unknown potential to impact the effectiveness and sustainability of chemotherapeutic approaches to disease transmission and control.

Author Notes

Address correspondence to Michael Cappello, Department of Pediatrics, Yale School of Medicine, P.O. Box 208081, New Haven, CT 06520. E-mail: michael.cappello@yale.edu

Financial support: This study was supported by the National Institutes of Health (AI099623; AI132452); Yale University (Wilbur Downs International Research Fellowship, Dean’s Research Fellowship); Jorge Paulo Lemann Foundation; University of Vermont (Brennan Summer Research Fellowship). The sponsors had no role in study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the article for publication.

Authors’ addresses: Ambrose R. Orr, Peter Suwondo, Santosh George, Lisa M. Harrison, and Michael Cappello, Department of Pediatrics, Yale School of Medicine, New Haven, CT, E-mails: ambrose.orr@gmail.com, peter.suwondo@aya.yale.edu, santosh.george@yale.edu, lisa.harrison@yale.edu, and michael.cappello@yale.edu. Josephine E. Quagraine and Michael D. Wilson, Department of Parasitology, Noguchi Memorial Institute for Medical Research, University of Ghana, Legon, Ghana, E-mails: jquagraine2@gmail.com and mwilson@noguchi.ug.edu.gh. Fabio Pio Dornas, Department of Pharmacy, School of Pharmaceutical Sciences, University of São Paulo, Ribeirao Preto, Brazil, E-mail: fabiopiod154@gmail.com. Benjamin Evans and Adalgisa Caccone, Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT, E-mails: benjamin.evans@yale.edu and adalgisa.caccone@yale.edu. Debbie Humphries, Department of Public Health, Yale University, New Haven, CT, E-mail: debbie.humphries@yale.edu.

These authors contributed equally to this work.

Save