The Effectiveness of a Pyriprole \(125 Mg/Ml\) and a Metaflumizone \(150
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Article available at http://www.parasite-journal.org or http://dx.doi.org/10.1051/parasite/2008151093 THE EFFECTIVENESS OF A PYRIPROLE (125 MG/ML) AND A METAFLUMIZONE (150 MG/ML) COMBINED WITH AMITRAZ (150 MG/ML) SPOT-ON TREATMENT IN PREVENTING PHLEBOTOMUS PERNICIOSUS FROM FEEDING ON DOGS THOMAS C.*, ROQUES M.* & FRANC M.* Summary: Résumé : PYRIPROLE ET ASSOCIATION MÉTAFLUMIZONE-AMITRAZ : ÉTUDE DE L’ACTIVITÉ ANTI-GORGEMENT VIS-À-VIS DE PHLEBOTOMUS A controlled clinical trial was performed to assess the effectiveness PERNICIOSUS SUR LE CHIEN TRAITÉ PAR CES FORMULATIONS SPOT-ON of a pyriprole (125 mg/ml) and a metaflumizone (150 mg/ml) combined with amitraz (150 mg/ml) spot-on treatment Cet essai avait pour but d’étudier l’efficacité de deux spot-on (recommended dosage) in preventing adult female sandflies destinés au chien – pyriprole (125 mg/ml) et métaflumizone (Phlebotomus perniciosus) from feeding on dogs. Sandfly mortality (150 mg/ml) associée à l’amitraz (150 mg/ml) – sur les was also assessed. Twelve beagle dogs were used in the study. phlébotomes (effet létal et effet antigorgement). 12 chiens ont été Prior to treatment they were checked for their attractiveness to sand- utilisés. Ils ont été répartis en trois lots de quatre en fonction de flies, ranked accordingly to generate partner triplets of equivalent leur attractivité pour les femelles de phlébotomes. Un lot a été sensitivity to sandflies: four control dogs, four treated with the traité avec le spot-on au pyriprole, un lot avec la métaflumizone pyriprole and four with the metaflumizone spot-on. The dogs were associée à de l’amitraz, le dernier lot étant le lot témoin non challenged with 50 unfed adult female sandflies (8-10 days old), traité. Un et sept jours après le traitement, les chiens ont été in cages for one hour on Day 1 and Day 7. The sandflies were soumis pendant une heure à des infestations par 50 femelles à checked for blood feeding and mortality at one hour, 24 hours jeun de Phlebotomus perniciosus âgées de 8-10 jours. L’état de and 48 hours after exposure to the dogs. A very poor anti-feeding gorgement a été observé une heure après, la mortalité notée 24 effect (near 7 %) was seen on sandflies with the metaflumizone et 48 heures après chaque infestation. L’effet antigorgement est combined with amitraz and no antifeeding effect was seen with faible (7 %) pour la métaflumizone associée à l’amitraz et nulle pyriprole. The sandfly mortality effect as a result of exposure to pour le pyriprole. La mortalité entraînée par les deux produits est treated dogs was under 20 % for the two spot-on. The two d’environ 20 %. Ces deux formulations ne présentent pas d’intérêt formulations could not be proposed in a leishmaniosis prevention dans un programme de prévention de la leishmaniose. program. MOTS CLÉS : métaflumizone, pyriprole, Phlebotomus, prévention phlébotome, KEY WORDS : metaflumizone, pyriprole, Phlebotomus, sandflie control, anti- antigorgement, chien, spot-on. feeding, dog, spot-on. eltamethrin and permethrin are used to prevent It appears interesting for the user to know in leish- culex and sandflies from feeding on dogs maniosis area if these new formulations are able to pro- D(Killick-Kendrick et al., 1997; Xiong et al., tect dogs from sand flies biting. 1994; Xiong et al., 1995). Products with these active ingredients have the claim “antifeeding effect on culex and sandflies” and also the claim “treatment and pre- MATERIALS AND METHODS vention of fleas and ticks infestation”. Two new formulations with new active ingredient DOGS (pyriprole alone, a phenylpyrazole insecticide and metaflumizone, a semicarbazone insecticide combined welve adult male and female Beagle dogs, wei- with an acaricide the amitraz) used in spot-on appea- ghing between 8.5 and 12.1 kg were housed red on the market. The claims are similar “treatment Tindividually in stainless steel cages in an envi- and prevention of fleas and ticks infestation”. The for- ronmentally controlled room, fed a commercial dog mulation is applied on the skin (one point), then the food with water supplied ad libitum. All dogs were active ingredient enters the lipid layer of the skin and identified by ear tattoo. After two weeks acclimatiza- spread rapidly across the all surface of the dog. tion and before treatment, all were challenged with unfed adult female sandflies, ranked according to the * UMR 181 Physiopathologie et Toxicologie Expérimentales, INRA- number of sandflies biting then into four partner tri- ENVT. École Nationale Vétérinaire, 23, chemin des Capelles, 31076 plets of approximately equal sensitivity and randomly Toulouse Cedex, France. allocated to one of the three groups (treated with pyri- Correspondence: Michel Franc. Tel.: 33 (0)5 61 19 38 73 – Fax: 33 (0)5 61 19 39 71. prole spot-on, treated with metaflumizone-amitraz spot- E-mail: [email protected] on or left untreated) using a random ABC sequence. Parasite, 2008, 15, 93-96 Note de recherche 93 THOMAS C., ROQUES M. & FRANC M. None of the dogs had received any antiparasitic treat- and classified (engorged or unengorged). The sandflies ment, which had ectoparasiticidal activity within two were fed on sugar-water and checked for mortality at months of the start of acclimatization. 24 and 48 hours after exposure to the dog. Dogs were observed regularly throughout the study. DATA ANALYSIS SANDFLIES MAINTENANCE AND SUPPLY The data collected at each time point were: number of Phlebotomus perniciosus (Diptera, Psychodidae), ori- live and engorged sandflies, number of live and un- ginally sourced from Lisbona (Dr Carlos PIRES-Por- engorged sandflies, number of dead and engorged sand- tugal), were cultured at ENVT using a 5-week egg to flies, number of dead and un-engorged sandflies. Effects adult cycle. 50 adult females 8-10 days old were used were assessed on a group basis (total numbers per treat- to challenge each dog. ment group). The statistical unit was the individual dog. The mortality and the anti-feeding rates were deter- EXPERIMENTAL PROCEDURE mined for each treatment group at each occasion of The dogs were treated with the unit label dose of each exposure of the dogs to sandflies (at 24 hours and at formulation respecting the laboratories recommenda- 48 hours after exposure) as follows: tions. For each product, the unit label dose of solu- total number of dead sandflies = mortality rate tion was applied topically by first parting the hair on total number of recovered sandflies the neck between the shoulders and applying the for- total number of unfed sandflies mulation directly on a single spot on the skin. = anti-feeding rate Dogs of the group A were untreated (Control group); total number of recovered sandflies dogs of group B were treated with a pyriprole spot-on The mortality effect and the anti-feeding effect of the ® ® (Prac-tic 137.5 mg or Prac-tic 275 mg depending of the treatment was calculated for each time point of assess- weight of the dog, Novartis) at the dosage of 13.7- ment by the following formula: 24.9 mg/kg; the dogs of group C were treated with a metaflumizone-amitraz spot-on (PromerisDuo 499.5 mg + (mortality rate in treated dogs)- ® (mortality rate in untreated dogs) 499.5 mg Fort Dodge) at the dosage of 39-46 mg/kg = mortality effect of each active ingredient. 1-(mortality rate in untreated dogs) The sandfly challenge assessment cages (60 cm × (anti-feeding rate in treated dogs)- × 40 cm 50 cm) were constructed from sandflies net- (anti-feeding rate in untreated dogs) = anti-feeding effect ting mounted on a wooden frame and placed in envi- 1-(anti-feeding rate in untreated dogs) ronmentally controlled rooms. Fifty unfed female sand- flies were placed in each of these four hours before Statistical analyses: comparisons between groups were introducing the dogs, which were immobilised using performed using the non parametric test Kruskal-Wallis ketamine (Imalgene® 9 mg/kg, Merial) and medetomi- (p < 0.05) dine (Domitor® 4 µg/kg, Pfizer) anaesthesia throughout the one hour exposure to the sand flies. Three partner pairs were challenged with sandflies at RESULTS any one time and all challenge assessments were done in virtual darkness except for some borrowed light to he results (Fig. 1) show that the experimental enable the dogs to be observed throughout the chal- infestations with Phlebotomus perniciosus were lenge period. Tsuccessful, with a low percentage of mortality At the end of the exposure period the dogs were (4 to 18 %) of the insects exposed on untreated dogs. checked carefully for any sandflies remaining on them, Between 41 and 48 mosquitoes were steel alive 48 hours then removed from the cage and allowed to regain full after each exposure to the control dogs. consciousness. All the live female sandflies collected, The mortality effect 48 hours after exposure to the trea- observed and classified (engorged or unengorged) ted dogs was approximately 20 % on D1 and 10 % on and placed in holding cages for observation 48 hours. D7 with pyriprole and 16-11 % with metaflumizone. The dead present in the assessment cage were counted (Table I and Fig. 1). The mean number of alive female 24 hours 48 hours Day infestation Pyriprole Metaflumizone Pyriprole Metaflumizone D10 7.20 0.00 19.86 6.32 D10 2.10 3.60 9.21 11.13 Table I. – Sandflie mortality effect (%). Parasite, 2008, 15, 93-96 94 Note de recherche PREVENTING P. PERNICIOSUS FROM FEEDING ON DOGS a a aaabbaab 50 45 40 35 Fig. 1. – Mean number of alive female 30 Pyriprole sandflies. 25 Metaflumizone Groups with different super- scripts are statistically significant 20 Control at p < 0.05. 15 female sandflies 10 Groups with different superscripts are statistically significant at p < 0.05.