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      Laboratory evaluation of the efficacy of lotilaner (Credelio™) against Amblyomma cajennense ( sensu lato) infestations of dogs

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          Abstract

          Background

          The ixodid tick Amblyomma cajennense ( sensu lato) complex, widespread throughout South and Central America, is also present in Mexico, Texas and Florida. As a vector of Rickettsia rickettsii, and potentially of other pathogens, infestations with A. cajennense present a substantial health risk to humans, dogs and other mammals. Oral administration of lotilaner flavored chewable tablets (Credelio TM, Elanco) to dogs was previously shown to rapidly provide killing activity of infesting ticks. This study investigated lotilaner’s efficacy against A. cajennense ( s.l.).

          Methods

          Twenty purpose-bred Beagles (10 male and 10 female) were ranked by Day -5 burdens of nymphal A. cajennense ( s.l.) and randomized to either treatment with lotilaner or to a sham-treated control group. On Day 0, dogs were fed within approximately 30 min prior to oral lotilaner administration at as close as possible to 20 mg/kg, the minimum dose rate. For efficacy assessments, tick counts were completed 48 h post-treatment or 48 h after experimental challenge infestations with 200 nymphal A. cajennense ( s.l.) on Days -7, -2, 7, 14, 21 and 28.

          Results

          Tick infestations in the control group dogs ranged from a low of 43 to 95, with the average infestation remaining above 25% at each assessment, thereby meeting the requirement for efficacy comparison with the treated group. Lotilaner efficacy was 100% within 48 h post-treatment, and at nine days post-treatment. Efficacy was greater than 99% at all subsequent assessments through Day 30. No treatment-related adverse events were observed.

          Conclusion

          The results demonstrate that lotilaner, administered orally to dogs at a minimum dose of 20 mg/kg is well tolerated, provides rapid reduction of existing A. cajennense ( s.l.) tick infestations, and provides sustained residual protection for at least 30 days against subsequent infestation by A. cajennense ( s.l.).

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          The novel isoxazoline ectoparasiticide fluralaner: selective inhibition of arthropod γ-aminobutyric acid- and L-glutamate-gated chloride channels and insecticidal/acaricidal activity.

          Isoxazolines are a novel class of parasiticides that are potent inhibitors of γ-aminobutyric acid (GABA)-gated chloride channels (GABACls) and L-glutamate-gated chloride channels (GluCls). In this study, the effects of the isoxazoline drug fluralaner on insect and acarid GABACl (RDL) and GluCl and its parasiticidal potency were investigated. We report the identification and cDNA cloning of Rhipicephalus (R.) microplus RDL and GluCl genes, and their functional expression in Xenopus laevis oocytes. The generation of six clonal HEK293 cell lines expressing Rhipicephalus microplus RDL and GluCl, Ctenocephalides felis RDL-A285 and RDL-S285, as well as Drosophila melanogaster RDLCl-A302 and RDL-S302, combined with the development of a membrane potential fluorescence dye assay allowed the comparison of ion channel inhibition by fluralaner with that of established insecticides addressing RDL and GluCl as targets. In these assays fluralaner was several orders of magnitude more potent than picrotoxinin and dieldrin, and performed 5-236 fold better than fipronil on the arthropod RDLs, while a rat GABACl remained unaffected. Comparative studies showed that R. microplus RDL is 52-fold more sensitive than R. microplus GluCl to fluralaner inhibition, confirming that the GABA-gated chloride channel is the primary target of this new parasiticide. In agreement with the superior RDL on-target activity, fluralaner outperformed dieldrin and fipronil in insecticidal screens on cat fleas (Ctenocephalides felis), yellow fever mosquito larvae (Aedes aegypti) and sheep blowfly larvae (Lucilia cuprina), as well as in acaricidal screens on cattle tick (R. microplus) adult females, brown dog tick (Rhipicephalus sanguineus) adult females and Ornithodoros moubata nymphs. These findings highlight the potential of fluralaner as a novel ectoparasiticide. Copyright © 2014 Elsevier Ltd. All rights reserved.
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            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.

            These second edition guidelines, updated from the 2007 version (Marchiondo et al., 2007), are intended to assist the planning and conduct of laboratory and clinical studies to assess the efficacy of ectoparasiticides applied to dogs or cats for the purpose of treating, preventing and controlling flea and tick infestations. Major revisions to this second edition include guidelines on the assessment of systemic flea and tick products, an update of the geographical distribution of the common fleas and ticks species on dogs and cats, determination of flea and tick efficacy based on geometric versus arithmetic means with respect to geographic regulatory agencies, modification of tick categorization in the assessment of efficacy, expanded guidelines on repellency and anti-feeding effects, enhanced practical field study guidance, and considerations on the ranges of flea and ticks for infestations in laboratory studies. The term ectoparasiticide includes insecticidal and acaricidal compounds, as well as insect growth regulators. The range of biological activities from animal treatment that are considered include: repellency and anti-feeding effects, knockdown, speed of kill, immediate and persistent lethal effects, and interference with egg fertility and subsequent development of off-host life cycle stages. Information is provided on the selection of animals, dose determination, dose confirmation and field studies, record keeping, interpretation of results and animal welfare. These guidelines are also intended to assist regulatory authorities involved in the approval and registration of new topical or systemic ectoparasiticides, and to facilitate the worldwide adoption of harmonized procedures.
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              Detection of Rickettsia rickettsii in the tick Amblyomma cajennense in a new Brazilian spotted fever-endemic area in the state of Minas Gerais

              The present study evaluated rickettsial infection in Amblyomma spp. ticks collected in a farm in Coronel Pacheco, a Brazilian spotted fever (BSF) endemic area. A total of 78 A. cajennense and 78 A. dubitatum free-living adult ticks were collected and tested by polymerase chain reaction (PCR) targeting a fragment of the rickettsial gene gltA. Only one pool of three A. cajennense ticks showed the expected product by PCR. This pool was further tested by PCR using sets of primers targeting the rickettsial genes gltA, ompA, and ompB. All reactions yielded the expected bands that by sequencing, showed 100% identity to the corresponding sequences of the Rickettsia rickettsii gene fragments gltA (1063-bp), ompA (457-bp), and ompB (720-bp). The minimal infection rate of R. rickettii in the A. cajennense population was 1.28% (at least one infected tick within 78 ticks).The present study showed molecular evidence for the presence of R. rickettsii in A. cajennense from a BSF-endemic area in Coronel Pacheco, state of Minas Gerais. Although R. rickettsii has been previously reported infecting A. cajennense ticks in Brazil and other Latin American countries, the present study performed the first molecular characterization of R. rickettsii from the tick A. cajennense.
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                Author and article information

                Contributors
                p_lasmar@elanco.com
                murphy_martin_gerard@elanco.com
                nanchen_steve@elanco.com
                drake_jon_j@elanco.com
                katherinac@gmail.com
                deb_vet@hotmail.com
                pricardim@ig.com.br
                scott.fabio@gmail.com
                Journal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                1756-3305
                3 October 2018
                3 October 2018
                2018
                : 11
                : 537
                Affiliations
                [1 ]Elanco Animal Health, Av. Morumbi, 8264, São Paulo, SP 04703–002 Brazil
                [2 ]Elanco Animal Health, Mattenstrasse 24a, CH-4058 Basel, Switzerland
                [3 ]ISNI 0000 0004 0638 9782, GRID grid.414719.e, Elanco Animal Health, ; 2500 Innovation Way, Greenfield, IN 46140 USA
                [4 ]ISNI 0000 0001 2294 473X, GRID grid.8536.8, Rural Federal University of Rio de Janeiro, ; BR 465 km 7 UFRRJ, Seropédica, RJ Brazil
                Author information
                http://orcid.org/0000-0001-6203-0852
                Article
                3116
                10.1186/s13071-018-3116-x
                6171179
                30285898
                87bb8919-4726-4f03-a427-a1c677ed15f6
                © The Author(s). 2018

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 15 June 2018
                : 18 September 2018
                Funding
                Funded by: Elanco Animal Health (US)
                Categories
                Research
                Custom metadata
                © The Author(s) 2018

                Parasitology
                canine,ticks,isoxazoline,effectiveness,cayenne tick
                Parasitology
                canine, ticks, isoxazoline, effectiveness, cayenne tick

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