Assessment of afoxolaner efficacy against Otodectes cynotis infestations of dogs
© The Author(s). 2016
Received: 3 October 2016
Accepted: 1 December 2016
Published: 9 December 2016
The efficacy of a single 2.5 mg/kg dose of afoxolaner (NexGard®, Merial) against induced Otodectes cynotis infestations was assessed in eight afoxolaner-treated dogs, compared to eight untreated dogs.
After O. cynotis infestations were established and confirmed by otoscopic assessments in 16 dogs, all of the dogs were included in the study and allocated to two separate treatment groups. The first group of eight ear mite-infested dogs remained untreated, while afoxolaner was administered orally to the second group of dogs at the minimum recommended dose once on Day 0. Otoscopic assessments performed on all dogs (Days -7, -2, 14 and 28) confirmed the presence or absence of live mites throughout the study. No serious adverse events were recorded throughout the study, and no adverse events were likely related to the administration of NexGard.
By Day 28, seven out of eight untreated dogs were still infested with ear mites, while only two out of eight afoxolaner-treated dogs were infested, with one and four ear mites, respectively. On Day 28, the reductions of mite counts in the afoxolaner-treated group versus those of the control dogs were 98.5% based on geometric means, and 99.4% based on arithmetic means. Significantly fewer (P < 0.05) live mites were present in the afoxolaner-treated group than the untreated group on Day 28.
The results of this study demonstrated that a single oral administration of afoxolaner at the minimum recommended dose is highly effective (>98%) in treating dogs with induced O. cynotis infestations.
The ear mite Otodectes cynotis, the causative organism of otodectic mange , is a very common non-burrowing, surface-living  acarian which infests cats, dogs and many other carnivores. Otodectes cynotis is found all over the world and primarily infests the ear canals of its host; however, it has also been found on head, back and tail in heavy infestations [1, 2]. The irritation caused by the mites often results in the host scratching its ears and shaking its head, which can lead to lacerations associated with the affected ear(s) or the formation of an auricular hematoma. Secondary yeast and bacterial infections often occur in the presence of heavy ear mite infestations.
Transmission of ear mites occurs by direct contact with an infested host [1–3], most commonly with the transmission of the mites from mother to kittens or puppies before weaning. Clinical signs which can lead to the suspicion of otodectic mange include bilaterally pruritic otitis with dry, blackish-brown cerumen in the host’s ear canal; self-induced ear lesions; aural hematomas; and pruritic lesions on the face and neck. Diagnosis is usually made through otoscopic examination of the ear and microscopic examination of cerumen, which usually contains ear mites of all life stages (eggs, larvae, nymphs and adults). Additionally, skin scrapings can occasionally reveal ear mites . It is important to note that, if at least one pet in the household is infested with ear mites, all pets in the household need to be treated for ear mites, as the mites are highly contagious [1, 4].
Since O. cynotis is a very common parasite of dogs and cats, it is essential to find effective treatments for infested pets. Historically, treatment necessitated ear flushing and cleansing with a mild ceruminolytic agent. Typical treatments include mineral oil or acaricides (monosulfiram, permethrin, or amitraz [3, 4]) directly administered into the ear canal.
Sometimes treating the ears alone would not solve the issue. Often to achieve control one would also treat the whole body with an acaricide dip or topical treatment, as mites can infest other parts of the body and re-infest the ears . Later, systemic-topical treatments such as selamectin and imidacloprid/moxidectin spot on formulations were found to have good systemic activity against ear mites in the ear canal and on the body [2, 3, 5, 6].
Recently, a new class of insecticides/acaricides, the isoxazolines, has demonstrated excellent efficacy against O. cynotis and other similar parasites . Afoxolaner is an isoxazoline administered monthly to protect dogs against fleas and ticks (NexGard®) [7–9]. It is administered at a minimum dose of 2.5 mg/kg, and comparative studies have shown that monthly administrations of afoxolaner provide outstanding protection against fleas and ticks [10, 11]. Afoxolaner has also demonstrated to be highly efficacious against Sarcoptes scabiei  and Demodex canis [13, 14]. Based on these observations, the present study was undertaken to assess the efficacy of a single oral administration of afoxolaner (NexGard) against O. cynotis infestations in dogs.
Individual dog information/details, as assessed on Day -1
Group 1 (negative control)
Group 2 (afoxolaner-treated)
Body weight (kg)
Body weight (kg)
4DE 28 F
956 36 F
Arithmetic mean body weight in kg: 19.4
Arithmetic mean body weight in kg: 18.3
Study design and treatment
This study was a randomized, single center, controlled efficacy study, following a randomized block design, and all evaluations of efficacy were performed by personnel in blinded conditions. Each dog was an experimental unit. For each dog, a total of > 10 live mites in at least one ear, as well as the presence of ≥ 1 live mite in the other ear, was necessary for inclusion in the study. Dogs were enrolled as they qualified, thus Day 0 was not the same calendar day for all dogs.
Administration of NexGard (afoxolaner) at ~2.5 mg/kg to the afoxolaner-treated group (Group 2) on Day 0
Body weight (kg)
Actual final dose (mg/kg)
1 × 28.3 mg & 1 × 11.3 mg
1 × 28.3 mg & 2 × 11.3 mg
1 × 28.3 mg & 2 × 11.3 mg
4 × 11.3 mg
4 × 11.3 mg
5 × 11.3 mg
1 × 28.3 mg & 2 × 11.3 mg
1 × 28.3 mg & 2 × 11.3 mg
Otoscopic assessments were performed on all dogs on Days -7, -2, 14 and 28 to confirm the initial presence, and subsequent presence or absence, of live mites.
Ear flushing and mite counts
On Day 28, a qualitative assessment of the ear canals was performed prior to the flushing procedure to determine the presence of viable mites. This was followed by the quantitative assessment of ear mites by ear duct flushing, complete mite collection and count. The ear ducts were filled with an appropriate solution of a 5% aqueous solution of docusate sodium (3 to 4 ml per ear canal), then the ears massaged externally until sufficient melting of the ear duct content was observed. The melted solution for each ear was collected into a labeled collection container, followed by a saline flush of the ear duct into the same container. The contents of the container were drained into a 38 μm sieve, which was then rinsed with clean water immediately, and the contents for each ear were transferred separately into labeled bottles. This procedure was conducted on each ear until otoscopic examinations revealed that both ears were clean. The contents of each ear were collected, examined, and stored separately in the labeled bottles. All collected material was observed microscopically on the same day, and all observed live mites (adult and immature) were counted and recorded for each ear.
The primary efficacy criterion was the number of live mites collected from the NexGard-treated group on Day 28, compared to that of the untreated control group on the same day. The statistical unit was the individual animal. The average percent reduction in mite counts for each group was calculated using geometric means: Efficacy (%) against ear mites = 100 × (GMC – GMT)/GMC, where GMC is the geometric mean number of live mites in dogs in the untreated group and GMT is the geometric mean number of live mites in dogs in the treated group. The groups were compared using a one-way ANOVA (Proc GLM procedure in SAS) with an administration effect after a logarithmic transformation on the mite (count + 1) data, and by using an ANOVA (Proc GLM procedure in SAS) with an administration effect on untransformed mite count data.
Additionally, due to large variability in mite counts in the negative control group, a non-parametric alternative test (the Mann-Whitney U-test) was used to assess efficacy by comparing median values. SAS version 9.3 TS Level 1 M2  was used for all statistical analyses. Statistical significance for geometric mean and arithmetic mean assessments was declared at a two-sided P-value of 0.05.
Clinical examinations and health observations
None of the clinical signs (mild pruritus, intermittent head-shaking) associated with the mite infestations observed during any of the clinical examinations were deemed as serious or affecting overall health and well-being. The examining veterinarian carefully examined and judged all enrolled dogs to be fit to continue in the study, at each assessment throughout the study. No serious health issues were recorded during the study, with only mild signs consistent with O. cynotis infestation observed. No adverse events related to the administration of afoxolaner were observed.
Efficacy against Otodectes cynotis based on geometric mean and arithmetic median number of mites recovered from both Group 1 (negative control) and Group 2 (afoxolaner-treated) dogs
Group 1 mean
Group 2 mean (% efficacy)
Group 1 mean
Group 2 mean (% efficacy)
Otodectes cynotis is a commonly occurring surface-dwelling mite that can easily transfer between suitable hosts [1–3]. Since these mites can exist on infected hosts outside the ear canals [1, 2], in order to effectively treat existing infestations, or potentially prevent infestations, effective systemic acaricidal products should be utilized. Properly ascertaining the overall efficacy of antiparasitic products often requires multiple studies be performed. Typically, the first such study performed is a controlled laboratory study utilizing the minimum target dose, realizing that studies of different scales will follow, should the results indicate a potential for success.
This study was designed to assess the efficacy of the minimum dose of afoxolaner against existing infestations of O. cynotis in dogs under controlled, laboratory conditions. The present study demonstrates that treatment with a single oral dose (at or approaching the minimum recommended dosage of 2.5 mg/kg) of afoxolaner (NexGard® Chewables) resulted in a rapid reduction in ear mite numbers, suggesting that a single oral administration of NexGard is highly effective (> 98%) against O. cynotis infestations in dogs, as assessed 28 days post-administration. This is comparable to efficacy achieved when afoxolaner was tested against other mite species infesting dogs, namely Sarcoptes scabiei (100% efficacy based on mite counts)  and Demodex spp., with > 99% efficacy based on mite counts in a well-controlled study  and resolution of cases under field conditions .
The high level of efficacy achieved with a single, minimum dose of NexGard® (afoxolaner) in the present study would seem to justify additional work to gather further evidence of the treatment and control capabilities of afoxolaner against O. cynotis, also when considering high efficacy obtained against other mite species infesting dogs.
The authors would like to thank L. van Zyl and H. Jooste for the statistical analysis.
This clinical study was funded by Merial, Inc.
Availability of data and material
Data supporting the conclusions of this article are included within the article.
DC, JF, CdV, and AL designed the study protocol. JF, CdV and AL were responsible for the conduct of the study. DC and JC drafted the first version of the manuscript, which was substantially revised by all authors. All authors read and approved the final manuscript.
Doug Carithers is an employee of Merial, Inc., and Jordan Crawford is a contractor for Merial Inc. ClinVet, of which Josephus Fourie, Christa de Vos and Alta Lotriet are employees, is an independent South African Contract Research Organization contracted to conduct the study. All authors voluntarily publish this article and have no personal interest in these studies, other than publishing the scientific findings in which they have been involved via planning, initiating, monitoring, and conducting the investigations and analyzing the results.
Consent for publication
The design and conditions of this study were approved by the Merial Institutional Animal Care and Use Committee (IACUC) and the local Animal Welfare Ethics Committee in accordance with Good Clinical Practice by the European Agency for the Evaluation of Veterinary Medicinal Products (CVMP/VICH GL9, July 2000; CVMP/VICH GL19, July 2001).
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