Abstract
A possible synergistic effect of macrocyclic lactones’ (MLs) combination has been previously described against resistant gastrointestinal nematodes of cattle. In addition to synergism, drug-drug interactions between MLs can also result in additive or antagonistic effect, considering the different MLs pharmacokinetics, pharmacodynamics, and interactions with molecular mechanisms of resistance. Therefore, the aim of the current work was evaluated the effect of different MLs combinations against Haemonchus contortus. Infecting larvae of two isolates (one susceptible and one resistant to ivermectin) were used in the larval migration inhibition test. After estimating the half maximal effective concentration of abamectin (ABA), eprinomectin, (EPR), ivermectin (IVM), and moxidectin (MOX) for both isolates, combinations were delineated by a simplex-centroid mixture experiment, and the mixture regression analysis was applied to the special cubic model. A synergistic effect was found for the EPR + MOX against the susceptible isolate as well as the EPR + MOX, IVM + MOX, and ABA + EPR + IVM against the resistant isolate. An antagonistic effect of ABA + IVM + MOX was found against the susceptible isolate. For the susceptible isolate, a higher inhibition was found with greater proportions of EPR and lower proportions of the other drugs compared to the reference mixture. For the resistant isolate, inhibition greater than that of the reference mixture was found with higher proportions of IVM as well as lower proportions of the other drugs. The synergistic and antagonistic effects were dependent on the following: (a) parasite drug resistance profile, (b) the composition of the combination, and (c) the proportions used, with EPR and IVM exerting a greater impact on these effects.
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References
Alvinerie M, Dupuy J, Kiki-Mvouaka S, Sutra JF, Lespine A (2008) Ketoconazole increases the plasma levels of ivermectin in sheep. Vet Parasitol 157:117–122. https://doi.org/10.1016/j.vetpar.2008.06.017
Anderson N, Martin PJ, Jarret RG (1988) Mixtures of anthelmintics: a strategy against resistance. Aust Vet J 65:62–64. https://doi.org/10.1111/j.1751-0813.1988.tb07355.x
Ballent M, Lifschitz A, Virkel G, Sallovitz JM, Lanusse C (2007) Involvement of P-glycoprotein on ivermectin kinetic behaviour in sheep: itraconazole-mediated changes on gastrointestinal disposition. J Vet Pharmacol Ther 30:242–248. https://doi.org/10.1111/j.1365-2885.2007.00848.x
Ballent M, Canton C, Dominguez P, Bernat G, Lanusse C, Virkel G, Lifschitz A (2020) Pharmacokinetic-pharmacodynamic assessment of the ivermectin and abamectina nematodicidal interaction in cattle. Vet Parasitol 279:109010. https://doi.org/10.1016/j.vetpar.2019.109010
Bartram DJ, Leathwick DM, Taylor MA, Geurden T, Maeder SJ (2012) The role of combination anthelmintic formulations in the sustainable control of sheep nematodes. Vet Parasitol 186:151–158. https://doi.org/10.1016/j.vetpar.2011.11.030
Bath GF (2006) Practical implementation of holistic internal parasite management in sheep. Small Rumins 62:13–18. https://doi.org/10.1016/j.smallrumres.2005.08.006
Blackhall WJ, Prichard RK, Beech RN (2003) Selection at a gamma-aminobutyric acid receptor gene in Haemonchus contortus resistant to avermectins/milbemycins. Mol Biochem Parasitol 131:137–145. https://doi.org/10.1016/S0166-6851(03)00201-9
Borges FA, Cho HS, Santos E, Oliveira GP, Costa AJ (2007) Pharmacokinetics of a new long acting endectocide formulation containing 2.25% ivermectin and 1.25% abamectin in cattle. J Vet Pharmacol Therap 30:62–67. https://doi.org/10.1111/j.1365-2885.2007.00816.x
Borges FA, Silva HC, Buzzulini C, Soares VE, Santos E, Oliveira GP, Costa AJ (2008) Endectocide activity of a new long-action formulation containing 2.25% ivermectin +1.25% abamectin in cattle. Vet Parasitol 155:299–307. https://doi.org/10.1016/j.vetpar.2008.04.019
Borges DGL, Araújo MA, Carollo CA, Carollo ARH, Lifschitz A, Conde MH, Freitas MG, Freire ZS, Tutija JF, Nakatani MTM, Borges FA (2020) Combination of quercetin and ivermectin: in vitro and in vivo effects against Haemonchus contortus. Acta Trop 201:105213. https://doi.org/10.1016/j.actatropica.2019.105213
Brown DD, Siddiqui SZ, Kaji MD, Forrester SG (2012) Pharmacological characterization of the Haemonchus contortus GABA-gated chloride channel, Hco-UNC-49: modulation by macrocyclic lactone anthelmintics and a receptor for piperazine Vet Parasitol 185:201–209. https://doi.org/10.1016/j.vetpar.2011.10.006
Cornell J (3rd Ed.) (2002) Experiments with mixtures: designs, models, and the analysis of mixture data. John Wiley & Sons, Inc., New York. 649 pp.
Demeler J, Küttler U, El-Abdellati A, Stafford K, Rydzike A, Varady M, Kenyon F, Coles G, Höglunde J, Jackson F, Vercruysse J, von Samson-Himmelstjerna G (2010) Standardization of the larval migration inhibition test for the detection of resistance to ivermectin in gastrointestinal nematodes of ruminants. Vet Parasitol 174:58–64. https://doi.org/10.1016/j.vetpar.2010.01.032
Echevarria F, Armour JL, Duncan JL (1991) Efficacy of some anthelmintics on an ivermectin-resistant strain of Haemonchus contortus in sheep. Vet Parasitol 39:279–284. https://doi.org/10.1016/0304-4017(91)90044-V
Fissiha W, Kinde MZ (2021) Anthelmintic resistance and its mechanism: a review. Infect Drug Resist 14:5403–5410. https://doi.org/10.2147/IDR.S332378
Forrester SG, Prichard RK, Beech RN (2002) A glutamate-gated chloride channel subunit from Haemonchus contortus: expression in a mammalian cell line, ligand binding, and modulation of anthelmintic binding by glutamate. Biochem Pharm 63:1061–1068. https://doi.org/10.1016/S0006-2952(02)00852-3
Forrester SG, Beech RN, Prichard RK (2004) Agonist enhancement of macrocyclic lactone activity at a glutamate-gated chloride channel subunit from Haemonchus contortus. Biochem Pharmacol 67:1019–1024. https://doi.org/10.1016/j.bcp.2003.08.047
Gao D, Chundawat SPS, Krishnan C, Balan V, Dale BE (2010) Mixture optimization of six core glycosyl hydrolases for maximizing saccharification of ammonia fiber expansion (AFEX) pretreated corn stover. Bioresour Technol 101:2770–2781. https://doi.org/10.1016/j.biortech.2009.10.056
Heckler RP, Almeida GD, Santos LB, Borges DGL, Neves JPL, Onizuka MKV, Borges FA (2014) P-gp modulating drugs greatly potentiate the in vitro effect of ivermectin against resistant larvae of Haemonchus placei. Vet Parasitol 205:638–645. https://doi.org/10.1016/j.vetpar.2014.08.002
Hibbs RE, Gouaux E (2011) Principles of activation and permeation in an anion-selective Cys loop receptor. Nature 474:54–60. https://doi.org/10.1038/nature10139
Kaplan RM, Vidyashankar AN (2012) An inconvenient truth: global warming and anthelmintic resistance. Vet Parasitol 186:70–78. https://doi.org/10.1016/j.vetpar.2011.11.048
Kathawala RJ, Gupta P, Ashby CR Jr, Chen ZS (2015) The modulation of ABC transporter-mediated multidrug resistance in cancer: a review of the past decade. Drug Resist Updat 18:1–17. https://doi.org/10.1016/j.drup.2014.11.002
Kohler P (2001) The biochemical basis of anthelmintic action and resistance. Int J Parasitol 31:336–345. https://doi.org/10.1016/S0020-7519(01)00131-X
Kotze AC, Ruffell AP, Knox MR, Kelly GA (2014) Relative potency of macrocyclic lactones in in vitro assays with larvae of susceptible and drug resistant Australian isolates of Haemonchus contortus and H. placei. Vet Parasitol 203:294–302. https://doi.org/10.1016/j.vetpar.2014.04.005
Laing R, Kikuchi T, Martinelli A, Tsai IJ, Beech RN, Redman E, Holroyd N, Bartley DJ, Beasley H, Britton C, Curran D, Devaney E, Gilabert A, Hunt M, Jackson F, Johnston SL, Kryukov I, Li K, Morrison AA, Reid AJ, Sargison N, Saunders GI, Wasmuth JD, Wolstenholme A, Berriman M, Gilleard JS, Cotton JA (2013) The genome and transcriptome of Haemonchus contortus, a key model parasite for drug and vaccine discovery. Gen Bio 14:R88. https://doi.org/10.1186/gb-2013-14-8-r88
Lanusse C, Canton C, Virkel G, Alvarez L, Costa-Junior L, Lifschitz A (2018) Strategies to optimize the efficacy of anthelmintic drugs in ruminants. Trends Parasitol 34:664–682. https://doi.org/10.1016/j.pt.2018.05.005
Laothanachareon T, Bunterngsook B, Suwannarangsee S, Eurwilaichitr L, Champreda V (2015) Synergistic action of recombinant accessory hemicellulolytic and pectinolytic enzymes to Trichoderma reesei cellulase on rice straw degradation. Bioresour Technol 198:682–690. https://doi.org/10.1016/j.biortech.2015.09.053
Leathwick DM, Hosking BC, Bisset SA, McKay CH (2009) Managing anthelmintic resistance: is it feasible in New Zealand to delay the emergence of resistance to a new anthelmintic class? N Z Vet J 57(4):181–192. https://doi.org/10.1080/00480169.2009.36900
Lespine A, Martin S, Dupuy J, Roulet A, Pineau T, Orlowski S, Alvinerie M (2007) Interaction of macrocyclic lactones with P-glycoprotein: structure–affinity relationship. Europ J Pharm Scie 30:84–94. https://doi.org/10.1016/j.ejps.2006.10.004
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 2:58–75. https://doi.org/10.1016/j.ijpddr.2011.10.001
Lifschitz A, Virkel G, Sallovitz J, Imperiale F, Pis A, Lanusse C (2002) Loperamide-induced enhancement of moxidectin availability in cattle. J Vet Pharmacol Ther 25:111–120. https://doi.org/10.1046/j.1365-2885.2002.00396.x
Lifschitz A, Suarez VH, Sallovitz J, Cristel SL, Imperiale F, Ahoussou S, Schiavi C, Lanusse C (2010) Cattle nematodes resistant to macrocyclic lactones: comparative effects of P-glycoprotein modulation on the efficacy and disposition kinetics of ivermectin and moxidectin. Exp Parasitol 125:172–178. https://doi.org/10.1016/j.exppara.2010.01.009
Lloberas M, Alvarez L, Entrocasso C, Virkel G, Ballent M, Mate L, Lanusse C, Lifschitz A (2013) Comparative tissue pharmacokinetics and efficacy of moxidectin, abamectin and ivermectin in lambs infected with resistant nematodes: Impact of drug treatments on parasite P-glycoprotein expression. Int J Parasitol Drugs Drug Res 3:20–27. https://doi.org/10.1016/j.ijpddr.2012.11.001
Molento M, Prichard R (1999) Effects of the multidrug-resistance-reversing agents verapamil and CL 347,099 on the efficacy of ivermectin or moxidectin against unselected and drug-selected strains of Haemonchus contortus in jirds (Meriones unguiculatus). Parasitol Res 85:1007–1011. https://doi.org/10.1007/s004360050673
Palmiera A, Sousa E, Vasconcelos MH, Pinto MM (2012) Three decades of P-gp inhibitors: skimming through several generations and scaffolds. Curr Med Chem 19(13):1946–2025. https://doi.org/10.2174/092986712800167392
Portillo V, Jagannathan S, Wolstenholme AJ (2003) Distribution of glutamate-gated chloride channel subunits in the parasitic nematode Haemonchus contortus. J Comp Neurol 462:213–222. https://doi.org/10.1002/cne.10735
Rao VT, Siddiqui SZ, Prichard RK, Forrester SG (2009) A dopamine-gated ion channel (HcGGR3*) from Haemonchus contortus is expressed in the cervical papillae and is associated with macrocyclic lactone resistance. Mol Biochem Parasitol 166:54–61. https://doi.org/10.1016/j.molbiopara.2009.02.011
Rao VT, Accardi MV, Siddiqui SZ, Beech RN, Prichard RK, Forrester SG (2010) Characterization of a noveltyramine-gated chloride channel from Haemonchus contortus. Mol Biochem Parasitol 173:64–68. https://doi.org/10.1016/j.molbiopara.2010.05.005
Rao VTS, Forrester SG, Keller K, Prichard RK (2011) Localisation of serotonin and dopamine in Haemonchus contortus. Int J Parasitol 41:249–254. https://doi.org/10.1016/j.ijpara.2010.09.002
Raza A, Kopp SR, Jabbar A, Kotze AC (2015) Effects of third generation P-glycoprotein inhibitors on the sensitivity of drug-resistant and -susceptible isolates. Vet Parasitol 211:80–88. https://doi.org/10.1016/j.vetpar.2015.04.025
Reyes-Guerrero DE, Cedillo-Borda M, Alonso-Morales R, Alonso-Díaz M, Olmedo-Juárez A, Mendoza-de-Gives P, López-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. https://doi.org/10.1016/j.molbiopara.2020.111281
Ringstad N, Abe N, Horvitz HR (2009) Ligand-gated chloride channels are receptors for biogenic amines in C. elegans. Sci 325:96–100. https://doi.org/10.1126/science.1169243
Roberts FHS, O’Sullivan PJ (1950) Methods for egg counting and larval cultures for strongyles infesting the gastrointestinal tract of cattle. Aust J Agr Res 1:99–102. https://doi.org/10.1071/AR9500099
Shalaby HA (2013) Anthelmintics Resistance; How to Overcome it? Iranian J Parasitol 8(1):18–32.
Smith G (1990) A mathematical model for the evolution of anthelmintic resistance in a direct life cycle nematode parasite. Int J Parasitol 20:913–921. https://doi.org/10.1016/0020-7519(90)90030-Q
Sutherland IA (2015) Recent developments in the management of anthelmintic resistance in small ruminants - an Australasian perspective. N Z Vet J 63:183–187. https://doi.org/10.1080/00480169.2015.1019947
Ueno H, Gonçalves PC (4th Ed.) (1998) Manual para diagnóstico das helmintoses de ruminantes. UFBA:UFRGS: Jap Int Coop Ag, Tokyo, 143 pp.
van Wyk JA (2001) Refugia–overlooked as perhaps the most potent factor concerning the development of anthelmintic resistance. Onderstepoort J Vet Res 68(1):55–67.
Willis HH (1921) A simple levitation method for the detection of hookworm ova. Med J Australia 8:375–376. https://doi.org/10.5694/j.1326-5377.1921.tb60654.x
Wolstenholme AJ, Fairweather I, Prichard R, von Samson-Himmelstjerna G, Sangster N (2004) Drug resistance in veterinary helminths. Trends Parasitol 20:469–476. https://doi.org/10.1016/j.pt.2004.07.010
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The authors thank Professor Doctor Alessandro Francisco Talamini do Amarante for generously furnishing the isolate of H. contortus susceptible to ivermectin.
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All authors contributed to the study conception and design. Material preparation and data collection were carried out by Matheus Takemi Muchon Nakatani, Dyego Gonçalves Lino Borges, Mário Henrique Conde, Mariana Green de Freitas, Juliane Francielle Tutija, Vinícius Duarte Rodrigues, and Guilherme Henrique Reckziegel. Data analysis were performed by Matheus Takemi Muchon Nakatani, Carlos Alexandre Carollo, and Fernando de Almeida Borges. The first draft of the manuscript was written by Matheus Takemi Muchon Nakatani, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Figure 1S
Response optimization plots of combinations with synergistic effect according to optimal proportions of abamectin (ABA), eprinomectin (EPR), ivermectin (IVM) and moxidectin (MOX) in mixtures against susceptible (RsHco1) and resistant (FAMEZHco1) isolates. Current proportion in mixture (dashed line and values in red), estimated relative inhibition of larval migration (dashed line and valued [%] in blue -y), inhibition curve according to proportion (solid black line), lower limit (grey area, 50% for susceptible isolate and 60% for resistant isolate). (PNG 469 kb)
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Nakatani, M.T.M., Borges, D.G.L., Conde, M.H. et al. Synergism of macrocyclic lactones against Haemonchus contortus. Parasitol Res 122, 867–876 (2023). https://doi.org/10.1007/s00436-023-07790-x
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DOI: https://doi.org/10.1007/s00436-023-07790-x