Effectiveness of Two Topical Treatments

With a Combination
Fipronil/Amitraz/(S)-methoprene Against
Natural Infestations of Mites
(Sarcoptes scabiei var. canis) on Dogs.
Soila Gaxiola1
Joel Gaxiola1
Ascencion Perez1
Stephen Yoon2
Jennifer Irwin2
Lenaig Halos2
Frederic Beugnet²
Matthias Pollmeier2
Roberto Alva2
1
University of Sinaloa, School of Veterinary Medicine, Carretera Internacional,
Salida Sur, Km. 3.5 Oriente, Culiacan, Sinaloa, Mexico.
2
Merial, 3239 Satellite Blvd, Duluth, GA 30096-4640, USA

KEY WORDS: Sarcoptes scabiei var. Sarcoptes scabiei var canis as confirmed
canis; sarcoptic mange; fipronil/amitraz/(S)- by skin scrapings collected prior to alloca-
methoprene; curative efficacy tion, were randomly divided into two equal
groups. Dogs in Group 1 were not treated.
ABSTRACT
Dogs in Group 2 were treated on Days 0 and
The acaricidal efficacy of the novel topical 28. At Days 0 (pretreatment), 7, 14, 21, 28
combination of fipronil/amitraz/(S)-metho- (before second treatment), 41, and 56, four
prene (CERTIFECT®) was evaluated against skin scrapings of similar size were taken
Sarcoptes scabiei var canis when admin- from different sites with lesions consistent
istrated as topical solution to dogs natu- with sarcoptic mange. Lesion scores were
rally infested with the mites. A total of 24 assigned weekly at Day 0 (pretreatment), 7,
mixed-breed dogs of both sexes (10 males 14, 21, 28 (before second treatment), 35, 42,
and 14 females), aged from 0.3 to 5 years and 56, and silhouettes identifying the loca-
and weighing 3.2 to 20.2 kg, were studied in tion of the present lesions were completed.
this single-center, randomized, blinded, and The results showed that dogs treated with
controlled trial. Dogs, naturally infested with CERTIFECT® had significantly (p<0.05)

10 Vol. 11, No. 1, 2013 • Intern J Appl Res Vet Med.

lower mite counts than untreated control ani- dogs; nevertheless, some have limitations
mals at Days 14, 21, 41, and 56 at a percent in their use and some are associated with
reduction of 100%, 99.3%, 100%, and 100% adverse reactions (Folz, 1994; Griffin, 1993;
respectively. In addition, dogs treated with Plumb, 1995; Curtis, 1996; Wendelberger
CERTIFECT® had significantly (p<0.05) and Wagner; 1998; Bordeau et Hubert, 2000;
lower lesion scores than untreated control Curtis, 2004).
animals on Days 28, 35, 42, 49, and 56. CERTIFECT® is a novel spot-on formu-
INTRODUCTION lation which combines fipronil, amitraz and
Canine sarcoptic mange is a highly con- (S)-methoprene. The combination of fipronil
tagious, intensely pruritic parasitic skin and (S)-methoprene (FRONTLINE® Plus/
disease caused by infestation with the Combo) formulated for topical application
epidermal mite Sarcoptes scabiei var canis. to dogs is known to provide a broad spec-
The disease commonly affects domestic trum of activity against insects and acarids
dogs with a worldwide distribution (Curtis, (Pollmeier et al, 2001; Curtis, 2004; Mc-Call
1996). Over half of the dogs, and up to 50% et al, 2004; Tielemans et al, 2010), but it
of human companions, can develop skin le- has been shown recently that the addition
sions after having contact with infested dogs of amitraz to this combination potentiates
(Folz, 1984; Griffin, 1993; Ihrke, 1994). The significantly the acaricidal effects of fipronil
mite is transmissible to humans (seen in up (Pfister et al., 2011; Prullage et al, 2011.).
to 50% of cases), but because the mites can- The purpose of the present study was to con-
not breed in human skin, humans are a dead- firm the efficacy of CERTIFECT® against
end host (Guaguère and Beugnet, 2008). natural infection with Sarcoptes scabiei var
Additionally, S. scabiei var canis has been canis, when administered to dogs.
isolated from species other than domestic MATERIALS AND METHODS
and wild canids and has been established
Study design
experimentally on several mammal species.
This lack of host specificity has therapeutic This study was a randomized, blinded,
and public health implications. In theory, all controlled efficacy trial of a combination of
mammals in contact with an infested dog fipronil/(S)-methoprene and amitraz against
should be treated simultaneously to limit the natural infections of Sarcoptes scabiei var
canis. The experimental study unit was the
opportunities for cross- and re-infestation
individual dog, identified by name, and natu-
(Curtis, 2004).
rally infested with S. scabiei var canis as
There are various topical therapies confirmed by skin scrapings collected prior
(lime sulfur, amitraz, organophosphates) to allocation to the study. The morphologic
and systemic therapies (milbemycin oxime, characteristics of the agent were determined
moxidectin, selamectin) currently used in according to the key published by Baker
the treatment of S. scabiei in dogs: (1999).
• acaricide amitraz dips (Curtis, 2004) In total, 24 mixed breed dogs of mixed
• selamectin as topical treatment sex (10 males and 14 females), aged from
(Shanks et al., 2000) 0.3 to 5 years, and weighing 3.2 to 20.2 kg,
• moxidectin in topical combination were allocated to this study. The naturally-
with imidacloprid (Krieger et al., infested animals were obtained from private
2005) owners who consented to the animal’s
• the combination of metaflumizone participation in the study.
with the amitraz in a spot-on formula- The 24 Sarcoptes-infested dogs were
tion (Fourie et al., 2007). divided randomly into two equal groups.
All were reported as being generally Dogs in Group 1 were not treated. Dogs in
effective treatments for sarcoptic mange in Group 2 were treated on Days 0 and 28 with

Intern J Appl Res Vet Med • Vol. 11, No. 1, 2013. 11

the CERTIFECT® formulation of Fipronil/ mineral oil were placed on the curette prior
(S)-methoprene and amitraz concurrently scraping. The debris was removed from the
administrated at a combined dose of 1.07 collected sample with a 25-gauge needle
mL, 2.14 mL or 4.28 mL of total volume prior to placing the scraping in a Petri dish
Body Fipronil/ Amitraz Total containing 2 mL of mineral oil (one Petri
weight (S)-methoprene volume dish per dog per day of analysis). Between
(kg) (mL) (mL) (mL) each scraping, the curette was wiped with a
Up to 10.0 0.67 0.40 1.07 paper towel and thoroughly scrubbed with
10.1-20.0 1.34 0.80 2.14 alcohol, then wiped again.
20.1-40.0 2.68 1.60 4.28 The number of mites (live and dead) in
the entire sample was counted using a light
microscope.
depending on the dogs body weight: Lesion Score
The minimum doses delivered were Sarcoptic mange lesion scores were assigned
6.7 mg/kg of fipronil, 6.03 mg/kg of (S)- and their locations (if present) were noted on
methoprene, and 8 mg/kg of amitraz. For the an animal silhouette. To maintain consis-
treatment administration, the total volume tency, the same person evaluated each dog
was applied directly onto the skin on two at each time point. Evaluation of individual
separate spots placed on the midline of the dogs was conducted using a five-point scor-
neck, between the base of the skull and the ing system:
shoulder blades.
0 no skin lesions
No adverse events were observed dur-
1 areas with papules and/or small
ing observations conducted approximately
areas with alopecia, < 4 areas over the
hourly for 4 hours immediately after the
body
CERTIFECT® treatment administration.
2 areas with papules and/or small
In order to perform the mite counts and
areas with alopecia, > 4 areas over the
control the effectiveness of the topical treat-
body
ments against the mites, four skin scrapings
of similar size were taken from different 3 more than half of the body with
sites with lesions consistent with sarcoptic papule and/or alopecia, crusts present
mange on Days 0, 7, 14, 21, 28, 41, and 4 chronic infestations with crusts
56. In the absence of skin lesions, scrap- covering more than half of the body
ings were obtained from previously active surface, skin reactions including alo-
sites. The collections on Days 0 and 28 were pecia or erythema.
performed before treatments were applied to Data Analysis
Group 2. Also lesion scores were assigned Mite Counts
and their locations (if present) were identi- The counts of live mites from the skin scrap-
fied on the Lesion Score form on Days 0, 7, ings were summed for each animal for cal-
14, 21, 28, 35, 42, 49, and 56. The Lesions culation of geometric means for each group
Scores on Days 0 and 28 were evaluated at each time point. The total live mite counts
before treatments were applied to Group 2. were transformed to the natural logarithm of
Specification of Study Variables counts (count +1) for calculation of geomet-
Mite Counts ric means for each group at each time point.
Skin scrapings were collected and per- Arithmetic means also were calculated. Per-
formed as follow: For each sample, an area cent reduction from the control group mean
of one square centimeter in size was scraped was calculated for the treated group at every
with a curette until blood began to appear. post-treatment time point using the formula
To ensure the attachment of the epidermal [(C-T)/C] × 100, where C is the geometric
scrapings to the curette, one to two drops of mean for the control group (Group 1) and T

12 Vol. 11, No. 1, 2013 • Intern J Appl Res Vet Med.

is the geometric mean for the treated group study on the basis both of physical examina-
(Group 2). The treated group was compared tions and of mite counts confirming infesta-
to the control group using Friedman’s test. tion with S. scabiei var canis. Dogs treated
Lesion Score with ectoparasiticides (either topical or
Lesion scores were summarized for each systemic) within 3 months of the start of the
group and by time point and were analyzed study were excluded. Dogs were observed
by Cochran-Mantel-Haenszel test. All daily for any health changes from Day -7
testing was two-sided at the significance until the end of the trial.
level α = 0.05. Supplemental descriptive RESULTS AND DISCUSSION
statistics (standard deviation, minimum, and The mite counts and the lesion score during
maximum) also were calculated. Analyses the 56 days of trial are summarized in the
were performed using procedures of SAS® Table 1 and Table 2. The 24 animals allo-
version 9.1. cated in this study were representative of the
Animal Management target population since there are no reported
As animals were obtained from private breed, sex, or age predilections for sarcoptic
owners, an informed consent and agreement mange (Curtis, 2004). All dogs presented
form was completed and signed by each skin abnormalities and clinical signs consis-
owner before any study-related activities tent with sarcoptic mange infections, which
were performed. The animals were managed included alopecia, papules, dermatitis,
similarly and with due regard for their well- erythema, and crusts.
being. They were handled in compliance CERTIFECT® demonstrated a signifi-
with Merial Institutional Animal Care and cant acaricidal efficacy from Day 14 post-
Use Committee (IACUC) approvals and re- treatment in the treated dogs. Indeed, results
quirements of the local IACUC. All animals from the mite counts showed that the treated
were housed in the same manner, individu- group had significantly (p<0.05) lower mite
ally in pens under controlled environmental counts compared to the untreated control
conditions. Food and water were delivered group at Days 14, 21, 41, and 56 (Table
ad libitum. On Day 0, all dogs were deter- 1). The percent reduction in the number
mined to be suitable for inclusion in the of mites when compared to the untreated
Table 1. Results of Mite counts by Study Day

Study Group 1 : untreated Group 2: CERTIFECT® Reduction P-value2

Day control Geometric Mean (Arith- (%)
Geometric Mean1 metic Mean)
(Arithmetic Mean)

0 19.8 (45.5) 18.4 (29.1) NA NA

7 0.7 (3.1) 0.3 (0.5) 57.1 0.6547
14 3.5 (21.8) 0 (0) 100 0.0143
21 1.5 (3.1) 0.1 (0.1) 93.3 0.0143
28 1.8 (8.1) 0.2 (0.3) 88.9 0.4142
41 2.2 (13.6) 0 (0) 100 0.0253
56 4.7 (40.8) 0 (0) 100 0.0143

1 Based on transformation to ln(count+1). There were 11 animals (one was euthanized on Day 5) in the untreated
group and 12 animals in the treated group.
2 Probability value from Friedman’s test.
NA = Not Applicable

Intern J Appl Res Vet Med • Vol. 11, No. 1, 2013. 13

control Group was 100%, 93.3%, 100%, and lesions typical of sarcoptic mange, because
100% respectively (Table 1). No mites could by the end of the study the dogs presented
be found in any CERTIFECT®-treated dogs almost no skin lesions anymore.
14 days following each treatments (ie, Days Efficacy of treatments against sarcoptic
14 and 41) and at the end of the study 28 mange are known to vary considerably; cure
days after the second treatment (ie,, Day 56). rates (based on mite counts and/or clinical
Furthermore the overall treatment suc- signs) can range from 60% to 100% (Wag-
cess was stated as a marked decrease in the ner and Wendelberger, 2000; Curtis, 2004;
severity and extent of mite-associated clini- Krieger et al, 2005; Fourie et al, 2006).
cal signs. The results of this study showed Despite treatments being reported as gener-
a significant therapeutic effect of CERTI- ally effective for sarcoptic mange control in
FECT, illustrated by the calculation of the dogs, some have limitations in their use and
Lesion Scores. Both groups of dogs pre- others are associated with adverse reactions
sented an average Lesion Score of 3 before (Folz, 1984; Griffin, 1993; Plumb, 1995;
treatment at the start of the study. From Day Curtis, 1996; Wendelberger and Wagner,
28 until the end of the experiment (ie, Day 1998; Bordeau and Hubert, 2000; Curtis,
56), the group of treated dogs presented sig- 2004). In this study, two treatments applied
nificant (p<0.05) lower Lesion Scores com- 1 month apart with a spot-on formulation
pared to the untreated control group (Table of fipronil/(S)-methoprene plus amitraz
2). Whereas the control group remained at a resulted in rapid reduction of mite counts
score ranging from 2 to 2.5 throughout the and in the cure of clinical signs. No adverse
56 days of the experiment, CERTIFECT®- events were observed following the treat-
treated dogs presented a substantial decrease ment administrations.
in the Lesion Scores ranging from 0 to 0.5 This study provided evidence that treat-
from Day 35 until the end of experiment. ment with CERTIFECT®, applied twice at
Importantly, the data indicated that treatment 28-day intervals, is highly effective against
resulted in a complete resolution of the skin Sarcoptes scabiei infection in dogs.
Table 2. Summary of Lesion Scores by Study Day
Study Group 1 : untreated control Group 2: CERTIFECT® P-value1
Day Median (Arithmetic Mean) Median (Arithmetic Mean)
0 3 (2.8) 3 (2.9) NA
7 3 (2.9) 3 (2.8) 1.0000
14 3 (2.9) 2.5 (2.7) 0.6547
21 3 (2.9) 2 (2.3) 0.1573
28 2 (2.5) 2 (1.5) 0.0348
35 2 (2.6) 0.5 (0.8) 0.0114
42 2 (2.6) 0.5 (0.8) 0.0114
49 2 (2.6) 0 (0.5) 0.0114
56 2.5 (2.6) 0 (0.1) 0.0027
1 Probability value from Cochran-Mantel-Haenszel test.
Lesion Score: 0 no skin lesions
1 areas with papules and/or small areas with alopecia, < 4 areas
2 areas with papules and/or small areas with alopecia, > 4 areas
3 more than half of the body with papules and/or alopecia, crusts present
4 chronic infestations with crusts covering more than half the body surface, skin reactions including alopecia or
erythema
NA = Not Applicable

14 Vol. 11, No. 1, 2013 • Intern J Appl Res Vet Med.

ACKNOWLEDGEMENT 12. Bordeau W. and Hubert B.: Treatment of 36 cases of
canine Sarcoptes using a 0.25 % fipronil solution.
CERTIFECT® and FRONTLINE® are Veterinary Dermatology, 2000, 11 (Suppl. 1): 27.
registered trademarks of Merial; all other 13. Pollmeier M., Pengo G., Barrick R.A. & Jeannin P.:
trademarks are the property of their respec- Efficacy of FRONTLINE® Spot-on against Ixodes
ricinus ticks in dogs. Proceedings of the 17th
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