Parasitology Research

, Volume 115, Issue 5, pp 1845–1851 | Cite as

Efficacy of oral afoxolaner plus milbemycin oxime chewables against induced infestations with Dermacentor reticulatus in dogs

  • Steffen RehbeinEmail author
  • Josephus J. Fourie
  • Christa de Vos
  • Andrew Anderson
  • Diane L. Larsen
  • Philippe Jeannin
Original Paper


The efficacy of afoxolaner plus milbemycin oxime (AFX + MO) combination chewables (NexGard Spectra®, Merial) and AFX single-entity chewables (NexGard®, Merial) against induced infestations with Dermacentor reticulatus ticks was evaluated in dogs. Thirty dogs were assigned to blocks of three animals each based on pre-allocation tick counts and were randomly allocated to one of three groups: untreated (control), treated with a combination of AFX + MO chewables to be as close as possible to the minimum effective dose of AFX + MO (2.5 + 0.5 mg per kg body weight), and treated with a combination of NexGard® chewables to be as close as possible to the minimum effective dose of AFX (2.5 mg per kg body weight). Treatments were administered orally once on day 0. Starting 2 days before treatment administration, each dog was infested with approximately 50 ticks weekly for six consecutive weeks. Live ticks were counted at ∼48 h post-treatment (removal count) and at ∼48 h (in situ counts) and ∼72 h (removal counts) following each post-treatment infestation. Treatment with both AFX + MO and NexGard® chewables rapidly eliminated the existing tick infestations (100 % efficacy) within 2 days following treatment administration. Weekly re-infestations were controlled for a minimum of 5 weeks with the efficacy ranging from 92.2 to 99.7 % based on ∼48 h post-treatment in situ counts and between 99.0 and 100 % based on ∼72 h post-treatment removal counts (p < 0.0001 at each occasion). This study demonstrated a high efficacy of both AFX + MO chewable and NexGard® chewable treatments against infestations of dogs with D. reticulatus ticks for at least 5 weeks. In addition, this study indicated no interference between the two compounds with respect to the acaricidal activity provided by AFX.


Dermacentor reticulatus Tick Afoxolaner Milbemycin oxime Dog 


Compliance of ethical standards

Conflict of interest

The work reported herein was funded by Merial, Inc., GA, USA. All authors are current/were employees or were contractors of Merial.


This document is provided for scientific purposes only. Any reference to a brand or trademark herein is for informational purposes only and is not intended for a commercial purpose or to dilute the rights of the respective owner(s) of the brand(s) or trademark(s).


  1. Beck S, Schreiber C, Schein E, Krücken J, Baldermann C, Pachnicke S, Gv S-H, Kohn B (2014) Tick infestation and prophylaxis of dogs in northeastern Germany: a prospective study. Ticks Tick-Borne Dis 5:336–342CrossRefPubMedGoogle Scholar
  2. Beugnet F, Halos L, Larsen D, Labuschagné M, Erasmus H, Fourie J (2014) The ability of an oral formulation of afoxolaner to block the transmission of Babesia canis by Dermacentor reticulatus ticks to dogs. Parasites & Vectors 7:283CrossRefGoogle Scholar
  3. Bona M, Pangrácová L, Peťko B (2011) Kliešte (Ixodidae) parazitujúce na psoch v rôznych klimatých oblastiach Slovenska a ich niektoré morfologické charakterstiky. Fourth International Scientific Conference – Infectious and Parasitic Diseases of Animals, 07–08 Sept 2011, Košice, Slovakia, Proceedings, pp 298–300Google Scholar
  4. Bugmyrin S, Hokkanen TJ, Romanova L, Bespyatova L, Fyodorv F, Burenkova L, Yakimova A, Ieshenko E (2011) Ixodes persulcatus (Schulze 1930) (Acari: Ixodidae) in eastern Finland. Entomol Fenn 22:268–273Google Scholar
  5. Bugmyrin SV, Bespyatova LA, Korotkov YS, Burenkova LA, Belova OA, Romanova LI, Kozlovskaya LI, Karganova GG, Ieshenko EP (2013) Distribution of Ixodes ricinus and Ixodes persulcatus ticks in southern Karelia (Russia). Ticks Tick-Borne Dis 4:57–62CrossRefPubMedGoogle Scholar
  6. Bullová E, Lukáň M, Stanko M, Peťko B (2009) Spatial distribution of Dermacentor reticulatus tick in Slovakia in the beginning of the 21st century. Vet Parasitol 165:357–360CrossRefPubMedGoogle Scholar
  7. Chitimia-Dobler L (2015) New data on the spatial distribution of the tick Dermacentor reticulatus in Romania. 25th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP), 16–20 Aug 2015, Liverpool, UK, Abstracts, p 474Google Scholar
  8. Claerebout E, Losson B, Cochez C, Casaert S, Dalemans A-C, de Cat A, Madder M, Saegerman C, Heyman P, Lempereur L (2013) Ticks and associated pathogens collected from dogs and cats in Belgium. Parasites & Vectors 6:183CrossRefGoogle Scholar
  9. Dautel H, Dippel C, Oehme R, Hartelt K, Schettler E (2006) Evidence for increased geographical distribution of Dermacentor reticulatus in Germany and detection of Rickettsia sp. RpA4. Int. J. Med. Microbiol. 296. Suppl 1:149–156Google Scholar
  10. Dumont P, Blair J, Fourie JJ, Chester ST, Larsen DL (2014) Evaluation of the efficacy of afoxolaner against two European dog tick species: Dermacentor reticulatus and Ixodes ricinus. Vet Parasitol 201:216–219CrossRefPubMedGoogle Scholar
  11. Duscher GG, Feiler A, Leschnik M, Joachim A (2013) Seasonal and spatial distribution of ixodid tick species feeding on naturally infested dogs from Eastern Austria and the influence of acaricides/repellents on these parameters. Parasites & Vectors 6:76CrossRefGoogle Scholar
  12. European Medicines Agency (2014) Committee for Medicinal Products for Veterinary Use (CVMP) assessment report for NexGard (EMEA/V/C/002729/0000) (12 December 2013, EMA/18910/2014). London, 21 ppGoogle Scholar
  13. European Medicines Agency (2015) Committee for Medicinal Products for Veterinary Use (CVMP) assessment report for NexGard Spectra (EMEA/V/C/003842/0000) (6 November 2014, EMA/695949/2014). London, 27 ppGoogle Scholar
  14. Evstaf’ev IL, Tovpinec NN (2002) Rhipicephalus sanguineus (Ixodidae) v Krymu: èkologičeskie i èpizootologičeskie aspekty. Vestnik Zool 36(4):85–91Google Scholar
  15. Fankhauser B, Hamel D, Dorr P, Reinemeyer CR, Crafford D, Bowman DD, Ulrich M, Yoon S, Larsen DL (2016) Efficacy of oral afoxolaner plus milbemycin oxime chewables against induced infections of gastrointestinal nematodes in dogs. Vet ParasitolGoogle Scholar
  16. Földvári G, Farkas R (2005) Ixodid tick species attaching to dogs in Hungary. Vet Parasitol 129:125–131CrossRefPubMedGoogle Scholar
  17. Földvári G, Márialigeti M, Solymosi N, Lukács Z, Majoros G, Kósa JP, Farkas R (2007) Hard ticks infesting dogs in Hungary and their infection with Babesia and Borrelia species. Parasitol Res 101(Suppl 1):S25–S34CrossRefGoogle Scholar
  18. Gilot B, Diop S, Laforge ML (1992) Dynamique spatiotemporelle des populations de Rhipicephalus sanguineus (Latreille, 1806) dans una cite HLM de Marseille. Acarologia 33:127–140Google Scholar
  19. Grandes AE (1986) Ticks of the province Salamanca (central/NW Spain): prevalence and parasitization intensity in dogs and domestic ungulates. Ann Parasitol Hum Comp 61:95–107Google Scholar
  20. Jaenson TGT, Tälleklint L, Lundqvist L, Olsen B, Chirico J, Mejlon H (1994) Geographical distribution, host associations, and vector roles of ticks (Acari: Ixodidae, Argasidae) in Sweden. J Med Entomol 31:240–256CrossRefPubMedGoogle Scholar
  21. Jongejan F (2015) Novel foci of Dermacentor reticulatus ticks infected with Babesia canis and Babesia caballi in the Netherlands and in Belgium. 25th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP), 16–20 Aug 2015, Liverpool, UK, Abstracts, p 390Google Scholar
  22. Jung M, Saito A, Buescher G, Maurer M, Graf J-F (2002) Chemistry, pharmacology and safety: milbemycin oxime. In: Vercruysse J, Rew RS (eds) Macrocyclic lactones in antiparasitic therapy. CABI Publ, Wallingford, pp 51–74CrossRefGoogle Scholar
  23. Karbowiak G (2014) The occurrence of the Dermacentor reticulatus tick—its expansion to new areas and possible causes. Ann Parasitol 60:37–47PubMedGoogle Scholar
  24. Kirkova Z, Iliev P, Visser M, Knaus M (2013) Survey of ectoparasites of dogs (Canis familiaris) in Bulgaria. 12th International Symposium on Ectoparasites of Pets (ISEP), 07–10 April 2013, Munich, Germany, Proceedings, p 68Google Scholar
  25. Knaus M, Rehbein S, Blair J, Richard-Mazet A, Visser M, Dollhofer D, Kley K, Lebon W, Anderson A, Larsen D, Jeannin P (2015) Field efficacy against tick and flea infestations and safety of afoxolaner plus milbemycin oxime chewables (NexGard Spectra®, Merial) in domestic dogs in Europe. 25th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP), 16–20 Aug 2015, Liverpool, UK, Abstracts, p 507Google Scholar
  26. Krčmar S, Ferizbegović J, Lonić E, Kamberović J (2014) Hard tick infestation of dogs in the Tuzla area (Bosnia and Herzegovina). Vet Arhiv 84:177–182Google Scholar
  27. Kunkle B, Daly S, Dumont P, Drag M, Larsen D (2014) Assessment of the efficacy of orally administered afoxolaner against Rhipicephalus sanguineus sensu lato. Vet Parasitol 201:226–228CrossRefPubMedGoogle Scholar
  28. Letendre L, Huang R, Kvaternik V, Harriman J, Drag M, Soll M (2014) The intravenous and oral pharmacokinetics of afoxolaner used as monthly chewable antiparasitic for dogs. Vet Parasitol 201:190–197CrossRefPubMedGoogle Scholar
  29. Letendre L, Harriman J, Drag M, Rehbein S (2016) The intravenous and oral pharmacokinetics of afoxolaner and milbemycin oxime when used as a combination chewable ecto-parasiticide and endectocide for dogs. J Vet Pharmacol TherapGoogle Scholar
  30. Maia C, Ferreira A, Nunes M, Vieira ML, Campino L, Cardoso L (2014) Molecular detection of bacterial and parasitic pathogens in hard ticks from Portugal. Ticks Tick Borne Dis 5:409–414CrossRefPubMedGoogle Scholar
  31. Marchiondo AA, Holdsworth PA, Fourie LJ, Rugg D, Hellmann K, Snyder DE, Dryden MW (2013) World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) second edition: Guidelines for evaluating the efficacy of parasiticides for the treatment, prevention and control of flea and tick infestations on dogs and cats. Vet Parasitol 194:84–97CrossRefPubMedGoogle Scholar
  32. Marotel MLS (2006) Tiques des carnivores domestiques en regions Rhône-Alpes, Auvergne, Limousin, Midi-Pyrénées, Aquitaine. Enquête de Juin à Décembre 2004. Thèse Doct Vét, Ecole Nat Vét ToulouseGoogle Scholar
  33. Mateescu R, Tudor P, Mateescu C (2011) A study regarding canine babesiosis in a veterinary clinic in Târgovişte-Dâmboviţa. Univ Agr. Sci Vet Med Bucharest, Sci Works. Ser C 57:279–285Google Scholar
  34. Medlock JM, Jameson LJ, Phipps LP (2011) Status of Dermacentor reticulatus in the UK. Vet Rec 168:386–387CrossRefPubMedGoogle Scholar
  35. Milutinović M, Radulović Z (2002) Ecological notes on ticks (Acari: Ixodidae) in Serbia (central regions). Acta Vet (Beograd) 52:49–58CrossRefGoogle Scholar
  36. Nijhof AM, Bodaan C, Postigo M, Nieuwenhuijs H, Opsteegh M, Franssen L, Jebbink F, Jongejan F (2007) Ticks and associated pathogens collected from domestic animals in the Netherlands. Vector-Borne Zoonotic Dis 7:1–11CrossRefGoogle Scholar
  37. Nolan TJ, Lok TB (2012) Macrocyclic lactones in the treatment and control of parasitism in small companion animals. Curr Pharmacol Biotechnol 13:1078–1094CrossRefGoogle Scholar
  38. Ogden NH, Cripps P, Davison CC, Owen G, Parry JM, Timms BJ, Forbes AB (2000) The ixodid tick species attaching to domestic dogs and cats in Great Britain and Ireland. Med Vet Entomol 14:332–338CrossRefPubMedGoogle Scholar
  39. Omeragic J (2011) Ixodid ticks in Bosnia and Herzegovina. Exp Appl Acarol 53:301–309CrossRefPubMedGoogle Scholar
  40. Otranto D, Dantas-Torres P (2010) Canine and feline vector-borne diseases in Italy: current situation and perspectives. Parasites & Vectors 3:2CrossRefGoogle Scholar
  41. Papadopoulos B, Morel PC, Aeschlimann A (1996) Ticks of domestic animals in the Macedonia region of Greece. Vet Parasitol 63:25–40CrossRefPubMedGoogle Scholar
  42. Papazahariadou MG, Saridomichelakis MN, Koutinas AF, Papadopoulos EG, Leontides L (2003) Tick infestation of dogs in Thessaloniki, northern Greece. Med Vet Entomol 17:110–113CrossRefPubMedGoogle Scholar
  43. Paulauskas A, Radzijevskaja J, Karvelienė B, Grigonis A, Aleksandravičienė A, Zamokas G, Babickaitė L, Sabūnas V, Petkevičius S (2014) Detection and molecular characterization of canine babesiosis causative agent Babesia canis in the naturally infected dog in Lithuania. Vet Parasitol 205:702–706CrossRefPubMedGoogle Scholar
  44. Pavlović I, Milutinović M, Petković D, Terzin D, Terzin V (2002) Epizootiological research of canine babesiosis in the Belgrade district. J Protozool Res 12:10–15Google Scholar
  45. Petney TN, Skuballa J, Muders S, Pfäffle M, Zetlmeisl C, Oehme R (2012) The changing distribution patterns of ticks (Ixodida) in Europe in relation to emerging tick-borne diseases. In: Mehlhorn H (ed) Arthropods as vectors of emerging diseases. Springer, Heidelberg, New York, Dordrecht, London, pp 151–166CrossRefGoogle Scholar
  46. Rehbein S, Dorr P, Bowman DD, Crafford D, Kusi I, Postoli R, Yoon S, Chester ST, Dollhofer D, Visser M, Larsen DL (2016) Efficacy of afoxolaner plus milbemycin oxime chewables against naturally acquired intestinal nematodes in dogs. Vet ParasitolGoogle Scholar
  47. Richter SH, Eydal M, Skírnisson K, Ólafsson E (2013) Tick species (Ixodidae) identified in Iceland. Icland Agr Sci 26:3–10Google Scholar
  48. Salman M, Tarrés-Call J (2013) Ticks and tick-borne diseases. CABI Publ, WallingfordGoogle Scholar
  49. Santos-Silva MM, Beati L, Santos AS, De Sousa R, Núncio MS, Melo P, Santos-Reis M, Fonseca C, Formosinho P, Vilela C, Bacellar F (2011) The hard-tick fauna of mainland Portugal (Acari: Ixodidae): an update on geographical distribution and known associations with hosts and pathogens. Exp Appl Parasitol 55:85–121Google Scholar
  50. Shoop WL, Hartline EJ, Gould BR, Waddel ME, McDowell RG, Kinney JB, Lahm GP, Long JK, Xu M, Wagerle T, Jones GS, Dietrich RF, Cordova D, Schroeder ME, Rhoades DF, Benner EA, Confalone PN (2014) Discovery and mode of action of afoxolaner, a new isoxazoline parasiticide for dogs. Vet Parasitol 201:179–189CrossRefPubMedGoogle Scholar
  51. Široky P, Kubelová M, Bednář M, Modrý D, Hubálek Z, Tkadlec E (2011) The distribution and spreading pattern of Dermacentor reticulatus over its threshold area in the Czech Republic—how much is range of this vector expanding? Vet Parasitol 183:130–135CrossRefPubMedGoogle Scholar
  52. Smith FD, Ballantyne R, Morgan ER, Wall R (2011) Prevalence, distribution and risk associated with tick infestation of dogs in Great Britain. Med Vet Entomol 25:377–384CrossRefPubMedGoogle Scholar
  53. Snyder DE, Wiseman S (2012) Dose confirmation and non-interference evaluations of the oral efficacy of a combination of milbemycin oxime and spinosad against the dose limiting parasites, adult cat flea (Ctenocephalides felis) and hookworm (Ancylostoma caninum), in dogs. Vet Parasitol 184:284–290CrossRefPubMedGoogle Scholar
  54. Sréter T, Széll Z, Varga I (2005) Spatial distribution of Dermacentor reticulatus and Ixodes ricinus in Hungary: evidence for change? Vet Parasitol 128:347–351CrossRefPubMedGoogle Scholar
  55. Tielemans E, Lebon W, Dumont P, Genchi M, Jeannin P, Larsen D (2015) Efficacy of oral afoxolaner plus milbemycin oxime chewable (NexGard Spectra®, Merial) to prevent heartworm disease in dogs after inoculation with third stage larvae of Dirofilaria immitis. 25th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP), 16–20 Aug 2015, Liverpool, UK, Abstracts, p 512Google Scholar
  56. Torina A, Khoury C, Caracappa S, Maroli M (2006) Ticks infesting livestock on farms in western Sicily, Italy. Exp Appl Acarol 38:75–86CrossRefPubMedGoogle Scholar
  57. Tudor P, Brăslaşu DE, Fernoagă C (2010) Study on natural infestation with hard ticks on dogs in Bucharest. Univ Agr Sci Vet Med Bucharest, Sci Works, Ser C 56:459–463Google Scholar
  58. Xhaxhiu D, Kusi I, Rapti D, Visser M, Knaus M, Lindner T, Rehbein S (2009) Ectoparasites of dogs and cats in Albania. Parasitol Res 105:1577–1587CrossRefPubMedGoogle Scholar
  59. Zygner W, Wędrychowicz H (2006) Occurrence of hard ticks in dogs from Warsaw area. Ann Agric Environ Med 13:355–359PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Steffen Rehbein
    • 1
    Email author
  • Josephus J. Fourie
    • 2
  • Christa de Vos
    • 2
  • Andrew Anderson
    • 3
  • Diane L. Larsen
    • 3
  • Philippe Jeannin
    • 4
  1. 1.Merial GmbH, Kathrinenhof Research CenterRohrdorfGermany
  2. 2.ClinVet International (Pty) Ltd.BloemfonteinRepublic of South Africa
  3. 3.Merial, Inc.DuluthUSA
  4. 4.Merial S.A.S., Centre de Recherche de Saint-VulbasSaint-VulbasFrance

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