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Insect growth regulators (IGRs) have been playing a major role in the effective management of a range of stored product insect pests including species that have developed resistance to major groups of insecticides, such as organophosphates (OPs) and synthetic pyrethroids (SPs). In the present study, we evaluated the efficacy of S-methoprene alone and in combination with piperonyl butoxide (PBO), an adjuvant component of insecticides for synergy, against two strains, Lab-S (susceptible) and Met-R (Methoprene resistant) of an economically important pest species, the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae). Adults of both Lab-S and Met-R strains were exposed to wheat treated with multiple concentrations of S-methoprene ranging from 0.001 to 0.01 and 10 to 60 mg/kg, respectively, alone and in combination with PBO. A variety of concentrations, including 0.27, 0.53, 0.80, and 1.07 g/kg, were evaluated for PBO. Mortality of adults and percent reduction in progeny were assessed after 14 and 65 days of treatment, respectively. As anticipated, the adult mortality rates of both strains were not significantly affected by S-methoprene alone. However, the number of progeny produced confirmed that the Met-R strain exhibited a high level of resistance to S-methoprene alone, with over 50 F1 progeny adults surviving in wheat treated with the maximal rate, 10 mg/kg. In contrast, the toxicity of S-methoprene was increased against the same resistant strain (Met-R), by 0.80 or 1.07 g/kg of PBO in combination treatment, resulting in a significant reduction in progeny numbers (25 adults per container). Although the tested concentrations of S-methoprene and PBO were well above the currently registered rate globally, our results highlight the fact that PBO enhances the toxicity of S-methoprene to some extent, reaffirming that the mode of action of the latter involves the inhibition of mixed-function oxidases (MFOs) and highlights the need for further research into developing potential binary or triplet formulations containing these two active ingredients (AIs)., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Mosquitoes and mosquito-borne illnesses significantly impact public health and human well-being. To address this concern, environmentally compatible larvicides have become a critical component of integrated mosquito management. However, the number of available larvicides is at a historical low. Currently, larvicides that harness microbials and insect growth regulators account for most products. Screening of new active ingredients (AIs) or improvement of existing AIs is thus necessary to augment the capacity for mosquito control. S-methoprene possesses a similar molecular structure and identical function to mosquito juvenile hormone and has been one of the main targets for research and development. The efficacy and safety of S-methoprene have been well documented since the late 1960s, and numerous products have been commercialized to combat pests of economic importance. However, S-methoprene is vulnerable to environmental factors that lead to its degradation, which has created challenges in formulation development, particularly where extended efficacy is desired. A derivative of S-methoprene, namely S-methobutene, with molecular modification has become available. This derivative has demonstrated an enhanced activity of inhibition of emergence (IE) against species across the Aedes, Anopheles, and Culex genera at IE10, IE50, and IE90. Furthermore, S-methobutene consistently outperformed S-methoprene during a 120-day aging process against the southern house mosquito Cx. quinquefasciatus, where the IE% in S-methobutene was significantly higher than that in S-methoprene on most aging intervals. The former had significantly longer residual activity than the latter. The potential of S-methobutene for further development and application is discussed in consideration of its enhanced activity and stability., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Eprinomectin and praziquantel, nematodicide and cestodicide compounds, are both combined with the insecticide and acaricide compounds fipronil and (S)-methoprene in Frontline ® Protect/Broadline ® , or esafoxolaner in NexGard ® Combo. These topical feline endectoparasiticide products were tested for efficacy against fleas and intestinal helminths in a field trial in Brazil. Flea- and/or helminth-infested domestic cats were treated twice at a monthly interval following label instructions: 160 cats with Frontline ® Protect/Broadline ® and 165 cats with NexGard ® Combo. The flea and intestinal helminth infestations were evaluated using comb counts and copromicroscopy, respectively before first treatment for baseline value, then 9 and 30 days after each treatment for fleas, and 9 days after each treatment for helminths. Multiparasitism was very frequent at baseline, as amongst the 325 included cats, 295, 280, 86 and 93 cats were at least infested with Ctenocephalides fleas, Ancylostoma, Toxocara and Dipylidium caninum, respectively. Efficacies were calculated by comparing the geometric means at baseline and at post-treatment timepoints for each parasite genus/species. Inclusive of both products and of all evaluation timepoints, the Ctenocephalides, Ancylostoma, Toxocara and D. caninum efficacies were at least 98.3%, 99.8%, 99.8% and 96.3%, respectively. No adverse reactions were observed, except for a few instances of mild, transient, and self-resolving hypersalivation occurring on the day of treatment in both groups. This field trial demonstrated high-level efficacy of Frontline ® Protect/Broadline ® and NexGard ® Combo against major parasites of cats in Brazil., (© A.A. Cutolo et al., published by EDP Sciences, 2022.)

Background: The lesser grain borer, Rhyzopertha dominica is a serious pest of stored grains. Fumigation and contact insecticides play a major role in managing this pest globally. While insects are developing genetic resistance to chemicals, hormonal analogues such as s-methoprene play a key role in reducing general pest pressure as well as managing pest populations that are resistant to fumigants and neurotoxic contact insecticides. However, resistance to s-methoprene has been reported in R. dominica with some reports showing a remarkable high resistance, questioning the use of this compound and other related analogues in grain protection. The current study attempts to identify possible molecular mechanisms that contribute in resistance to s-methoprene in R. dominica., Results: Transcriptome analysis of resistant and susceptible strains of this pest species identified a set of differentially expressed genes related to cytochrome P450s, indicating their potential role in resistance to s-methoprene. Laboratory bioassays were performed with s-methoprene treated wheat grains in presence and absence of piperonyl butoxide (PBO), a cytochrome P450 inhibitor. The results indicate that PBO, when applied alone, at least at the concentration tested here, had no effect on R. dominica adult emergence, but has a clear synergistic effect to s-methoprene. The number of produced progeny decreased in presence of the inhibitor, especially in the resistant strain. In addition, we also identified CYP complement (CYPome) of R. dominica, annotated and analysed phylogenetically, to understand the evolutionary relationships with other species., Conclusions: The information generated in current study suggest that PBO can effectively be used to break resistance to s-methoprene in R. dominica.

This work evaluated tick and flea burdens and the efficacy of a single topical application of 10% fipronil + 9% (S)-methoprene spot-on against experimental infestations of Rhipicephalus sanguineus sensu lato and Ctenocephalides felis felis on dogs that were submitted to regular baths for 56-days post-treatment. Four treatments (n = 24) were evaluated: no chemical treatment and no bathing (T01); no chemical treatment + bathing (T02); chemical treatment and no bathing (T03) and chemical treatment + bathing (T04). Dogs were infested with adult ticks and fleas seven days pre-treatment (-7). The ectoparasites were then counted and removed on day -5, followed by a new infestation on day -2 and treatment on day 0. Ticks and fleas were then counted without removal on day 1 and counted with removal on day 2. The dogs were then repeatedly infested with both ectoparasites on days 5, 12, 19, 26, 33, 40, 47 and 54 post-treatment. Baths and ectoparasite counts with removal were performed every 48 h before and after experimental infestation, respectively. Animals of T04 had lower (P ≤ 0.05) tick burden than animals from T01 and T02 from day 2 to day 28. For fleas, the counts varied among days of the study. Dogs from T03 presented lower (P ≤ 0.05) flea burden than dogs from T01 from day 1 until day 56. The same was true for T04 when compared to T01 from day 1 to day 42. Animals of T02 greater (P ≤ 0.05) flea burden than animals of T04 in all dates post treatment. In addition, dogs from T02 presented higher (P ≤ 0.05) flea burden than T01 from day 7 to day 28 and from day 49 to day 56. In sum, the efficacy of the spot-on formulation used declined over the time against both ectoparasites. The efficacy for ticks ranged from 0.0 to 96.5% and 0.0 to 98.52% for T03 and T04, respectively. While for fleas, the efficacy of the formulation ranged from 67.41 to 100% for T03 and 40.18 to 100% for T04. So, bathing dogs that were not treated increased C. f. felis burden but not R. sanguineus s. l. burden. The residual efficacy of the product had a shorter duration against these ectoparasites for dogs that received subsequent bathing., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: This controlled laboratory study was designed to evaluate the efficacy of the 10 % imidacloprid/4.5 % flumethrin collar (Seresto®, Bayer Animal Health) against fleas (Ctenocephalides f. felis) on cats, when compared to fipronil (9.8 %w/w)/(s)-methoprene (11.8 % w/w) topical spot-on formulation (Frontline® Plus for Cats and Kittens, Merial)., Methods: Thirty cats were randomized into three groups of ten animals based on pre-treatment flea counts: Group 1: imidacloprid/flumethrin collar; Group 2: fipronil/(s)-methoprene topical spot-on and Group 3: non-treated controls. The imidacloprid/flumethrin collars were applied one time on Day 0, while the fipronil/(s)-methoprene spot-on was administered every 30 days from Day 0 through Day 210. Cats were infested with 100 fleas on study days 0, 7, 14, 29, 59, 89, 119, 149, 179, 209 and 239. All flea counts were conducted by combing to remove fleas on post-treatment days 2, 8, 15, 30, 60, 90, 120, 150, 180, 210 and 240., Results: The efficacy of the imidacloprid/flumethrin collar ranged from 98.2 to 100 % for eight months. The efficacy of fipronil/(s)-methoprene spot-on ranged from 68.2 to 99.9 %. Efficacy was < 85 % for fipronil/(s)-methoprene on Days 90, 150 and 210. The flea counts in both treatment groups were significantly fewer than those in the non-treated control group at every post-treatment study day (P < 0.0001). In addition, there were significantly fewer fleas in the imidacloprid/flumethrin collar group when compared to the fipronil/(s)-methoprene group on Days 90, 150 and 210 (P < 0.0001)., Conclusions: This study demonstrated that the imidacloprid/flumethrin collar (Seresto®, Bayer Animal Health) maintained excellent ( > 98.2 %) efficacy against fleas on cats for the entire 8 month study. Monthly applications of fipronil/(s)-methoprene (Frontline® Plus for Cats and Kittens, Merial) generally had high, but variable (68.2 to 99.9 %) efficacy over the course of the eight month study. Based on the very high residual efficacy achieved by the imidacloprid/flumethrin collar in this study, veterinarians should expect that this collar will control and eliminate existing flea infestations on cats and in their in-home premises as long as every flea infested host is treated.

Background: Efficacy of FRONTLINE TRITAK For Dogs (fipronil/(S)-methoprene/cyphenothrin, Merial, Inc., Duluth, GA) against Ixodes scapularis was evaluated in two separate, but concurrent laboratory studies., Methods: One day after topical treatment with placebo or active, dogs (n = 24) were infested with 50 unfed adult Ixodes scapularis ticks, with repeat infestations on Days 7, 14, 21 and 28. The number of live ticks was counted at 6 hours post-infestation in the first study (n = 12) and at 24 hours post-infestation in the second study (n = 12)., Results: Observed efficacies in study 1 were 93-99% at 6 hour assessments on Day 1 through Day 28 and in the second study, 98-100% at 24 hour assessments, occurring on Day 2 through Day 29., Conclusions: A single dose of FRONTLINE TRITAK For Dogs (fipronil/(S)-methoprene/cyphenothrin) (0.67 ml or 1.34 ml) prevented the establishment of a new infestation following treatment, as well as the repeated weekly re-infestations with Ixodes scapularis ticks, for 4 weeks.

Background: The efficacy of methoprene can vary with surface substrates, application methods and environmental conditions. The objectives of the present study were to examine the effects of temperature and grain type on the long-term persistence and efficacy of methoprene in controlling Rhyzopertha dominica from Guangzhou, China., Results: Methoprene applied at 1 mg kg(-1) caused >90% suppression of F1 adult progeny of R. dominica for 150-270 days. Temperature and grain type both influenced the long-term persistence of methoprene. Overall multivariate analysis of variance showed that the order of progeny reduction at different temperatures (°C) was: 24 > 28 > 32 > 36; the order of the progeny reduction on the different grains was: paddy > wheat and maize., Conclusion: The results of our experiments show the maximum effect of methoprene for R. dominica control at 24 °C on paddy., (© 2013 Society of Chemical Industry.)

Background: A recently launched topical ectoparasiticide containing fipronil, amitraz and S-methoprene has been associated with the development of an acantholytic pustular dermatitis similar to that of Promeris-triggered pemphigus foliaceus (PF)., Hypothesis/objectives: Our objectives were to describe the clinical, histological and immunological features of this PF-like cutaneous adverse drug reaction., Animals: Twenty-one dogs with a probable or definitive diagnosis of PF-like cutaneous adverse drug reaction were identified between May 2012 and February 2013., Material and Methods: Histology, direct and indirect immunofluorescence were employed to address the study objectives., Results: Most dogs were middle-aged or older (median, 9 years) and of large size (median, 23 kg). In six dogs (29%), the PF-like lesions remained confined to the site of application, while 15 dogs (71%) exhibited lesions at distant sites. One or two applications of the ectoparasiticide were sufficient to trigger PF-like lesions in seven (33%) and six (29%) dogs, respectively. Systemic signs were reported in nine dogs (43%), all with lesions extending to sites distant from application areas. Tissue-bound antikeratinocyte IgG was detected in the lesional epidermis of eight of 19 (42%) cases by direct immunofluorescence, while serum antikeratinocyte IgG was detected in 10 of 14 (71%) cases by indirect immunofluorescence. Autoantibodies were found to target canine desmocollin-1 in 11 of 14 dogs (79%), but not canine desmoglein-1, by indirect immunofluorescence on transfected cells. These immunological findings were similar in cases with localized and distant disease., Conclusions and Clinical Importance: This new topical ectoparasiticide containing fipronil, amitraz and S-methoprene is capable of triggering the development of an acantholytic pustular dermatosis that is a clinical, histological and immunological close match for Promeris-triggered PF and naturally occurring autoimmune PF in dogs., (© 2014 ESVD and ACVD.)

The efficacy of the treatment with Certifect ® (containing fipronil 6.26% w/v, amitraz 7.48% w/v, (S)-methoprene 5.63% w/v) applied topically was assessed in 18 dogs diagnosed with clinical generalized demodicosis. Three treatment regimens were compared over a 3-month period. Starting at Day 0, dogs were treated monthly (group 1) or every two weeks (group 2) with the combination of fipronil, amitraz, and (S)-methoprene or with monthly topical applications of the combination of amitraz and metaflumizone (group 3, reference treatment). Clinical examinations including deep skin scrapings were performed every month in order to evaluate the resolution of clinical signs and the reduction in mite counts. On Day 84, the percentage reduction of mite counts in group 1 was 99.8%, whereas no Demodex canis could be detected in groups 2 and 3 (i.e. 100% parasitological efficacy). As a result of the Demodex mite count reduction, the skin condition of the dogs improved significantly in all groups. This study illustrates, that both monthly and bi-weekly treatments with Certifect were effective in treating dogs with generalized demodicosis over a 3-month period., (© J. Fourie et al., published by EDP Sciences, 2013.)

Flea infestations of pets continue to persist due to the lack of knowledge of flea biology and ecology. It is not unusual that pet owners believe regular hygiene, such as shampooing their dogs can replace regular insecticidal treatment. The objective of this study was to compare in a flea simulated environment, modelling exposure similar to that found in a home, that the use of regular physiological shampoo does not control fleas adequately when compared to a long acting topical formulation. Three groups of six dogs were formed: one untreated control group, one group treated monthly with the topical formulation of fipronil/(S)-methoprene, and a third group treated weekly with a hygiene shampoo. All dogs were infested with adult unfed Ctenocephalides felis fleas (200±5) on Days -28 and -21. Each animal's sleeping box was fitted with a plastic cup mounted to the inside roof of the box. The sleeping bench of each animal was covered with a carpet to accommodate flea development. The dogs were maintained in their kennels throughout the study. In order to maintain the environmental flea challenge, C. felis pupae (100±5) were placed in the plastic cup in each animal's sleeping box on Days -14, -7, 0, 7, 14, 21, 28, 35 and 42. The dogs were combed and fleas counted weekly on Days -1, 3, 10, 17, 24, 31, 38, 45, and 51. The fleas were placed immediately back on the dogs. On Day 60, fleas were counted and removed. Flea infestations in the untreated control group at each count averaged between 46.2 and 74.2 fleas throughout the study. The average number of fleas infesting dogs was significantly different (p<0.05) between the untreated and the two treatment groups and between the two treatment groups at all counts throughout the two months study (Days 3, 10, 17, 24, 31, 38, 45, 51 and 60). The efficacy was never below 99.1% in the fipronil/(S)-methoprene group, and efficacy in the shampoo group was never above 79.2%. Weekly shampooing in treatment group 3 was intentionally delayed after Day 42, to evaluate wether missing a weekly bath would affect the flea population. The Day 48 data indicate that forgetting or delaying a single weekly shampooing resulted in a clear increase in flea numbers and a significant decrease in efficacy from 68.2% to 34.8%. The fipronil(S)/methoprene treatment allowed a continuous control as demonstrated by the high efficacy against fleas, and also the number of flea-free dogs on seven of the nine weekly assessments, in spite of what was essentially a continuous flea challenge model.

Background: Two studies evaluating the efficacy of an imidacloprid/flumethrin collar (Seresto®, Bayer Animal Health, IVP), a deltamethrin collar (Scalibor®, MSD, CP1), a fipronil/(s)-methoprene spot-on (Frontline Combo®, Merial, CP2), a dinotefuran/pyriproxyfen/permethrin spot-on (Vectra 3D®, Ceva, CP3) and an amitraz/fipronil/(s)-methoprene spot-on (Certifect®, Merial, CP4/CP5) against repeated infestations with Rhipicephalus sanguineus and Ctenocephalides felis felis on dogs were conducted over periods of 226 days and 71 days respectively., Methods: The first study comprised 4 groups of treated dogs and one untreated control group, and the second 3 groups of treated dogs and one control group. Each group consisted of 8 dogs. All dogs were infested with ticks and fleas at regular intervals. Ticks were counted 6 h, 18 h or 48 h after infestations and fleas 24 h after infestations. Efficacies of the treatments were calculated by comparison with the untreated control groups using standard descriptive statistics., Results: The protective 48 h tick efficacy was 97.8% to 100% for the IVP (226 days), 69.3% to 97.4% for CP1 (170 days), 99.6% to 43.4% for CP2 (35 days) and 98% to 61.4% for CP3 (35 days).The protective 18 h tick efficacy was 98% to 99.6% for the IVP (71 days), 100% to 86.5% for CP4 (29 days), 100% to 72.8% for CP4 after re-treatment (35 days) and 98.8% to 54.3% for CP5 (35 days).The protective 6 h tick efficacy was 85.6% at Day 7 and 90.1% to 97.1% from Day 14 onwards for the IVP (70 days), 92.3% to 70.7% for CP4 (35 days), 97.5% to 65.2% for CP4 after re-treatment (35 days) and 95.1% to 51.8% for CP5 (35 days).The protective 24 h flea efficacy was 99.5/90.9% to 100% for the IVP (71/226 days), 66.7% to 83% for CP1 (170 days), 100% to 88.5% for CP2 (35 days), 100% to 73.3% for CP3 (35 days), 100% to 98.7% for CP4 (35 days), 100% to 87.5% for CP4 after re-treatment (35 days) and 100% to 79.5% for CP5 (35 days)., Conclusions: These data suggest that the long-term efficacy provided by a medicated collar that is effective, is a means to overcome the fluctuating efficacy of spot-on treatments resulting from a lack of pet owner re-treatment compliance, and consequently protect animals successfully against ectoparasites and probably vector-borne diseases.

Thirteen cats and 7 dogs living in 14 homes were treated topically with either a dinotefuran (22%, w/w)/pyriproxyfen (3.00%, w/w) (DP) or dinotefuran (4.95%, w/w)/pyriproxyfen (0.44%, w/w)/permethrin (36.08%, w/w) (DPP) topical spot-on, respectively. Twenty cats and 7 dogs living in 16 homes were treated topically with either a fipronil (9.8%, w/w)/(S)-methoprene (11.8%, w/w) or fipronil (9.8%, w/w)/(S)-methoprene (8.8%, w/w) topical spot-on (FM), respectively. All products were applied according to label directions by study investigators on day 0 and again between days 28 and 30. Flea populations on pets were assessed using visual area counts and premise flea infestations were assessed using intermittent-light flea traps on days 0, 7, 14, 21, 28-30, 40-45, and 54-60. A single application of the DP-DPP and FM formations reduced flea populations by 87.35% and 88.44%, respectively within 7 days. Following two monthly applications of either the DP-DPP or FM formulations, pet flea burdens were reduced by 95.24% and 95.47%, respectively. Flea numbers in the indoor-premises were also markedly reduced by days 54-60, with 98.05% and 96.15% reductions in intermittent-light flea trap counts in the DP-DPP or FM treatment groups, respectively., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The sustained speed of kill against Dermacentor reticulatus of two topical combinations, one containing fipronil/amitraz/(S)-methoprene and the other, imidacloprid/permethrin, was evaluated in dogs. Two treated groups and one untreated control group of eight adult Beagle dogs each were randomly formed based on pre-infestation rates and bodyweight. Each treatment was administered topically once on Day 0, according to the recommended label dose and instructions for use. All dogs were infested with 50 adult unfed D. reticulatus starting on Day 1, then weekly, for a total of five weeks. While most studies determine tick efficacy at 48 hours (h), in this study, all remaining ticks were counted and categorized 24 h following each infestation. The numbers of ticks (living or dead) that remained attached on treated dogs were compared to those on the control animals. The percent reduction of attached ticks (disruption of attachment) at 24 h on dogs treated with fipronil/amitraz/(S)-methoprene remained above 92% for four weeks. The reduction of attached ticks at 24 h on dogs treated with imidacloprid/permethrin did not reach 80% during the entire study. The number of ticks attached at 24 h was significantly (p<0.05) lower in the fipronil/amitraz/(S)-methoprene group than in the imidacloprid/permethrin group in assessments on Days 2, 15, 22, 29 and 36. When assessing efficacy based upon live ticks on treated versus control dogs, fipronil/amitraz/(S)-methoprene 24 h efficacy was above 95% for four weeks, decreasing to 77.8% at Day 36. The 24 h efficacy of imidacloprid/permethrin ranged from 56.2% to 86.7% through Day 29, never achieving 90% throughout the study. The 24-hour efficacy of fipronil/amitraz/(S)-methoprene was significantly (p<0.05) higher than imidacloprid/permethrin at all time points, including Day 36.

In the study reported here, the pioneer fipronil/(S)-methoprene topical product (Frontline® Plus, Merial Limited, Duluth, GA) was compared to the oral spinosad product (Comfortis® Elanco, Greenfield, IN) for efficacy against adult fleas and preventing egg production. The product presentations, doses and labelling were the one applicable in the USA. Using a standard protocol, 200 cat fleas of mixed sex were applied to dogs on Days 1, 7, 14, 21, 28, 35, and 42. Dogs were combed to remove fleas 24 hours post-infestation, the fleas were counted, collected, and then reapplied to each dog following completion of their respective count. At 48 hours post-infestation, comb counts were performed and fleas were removed. No fleas were collected from any dog in the fipronil/(S)-methoprene group at any 24 or 48 hours post-infestation assessment throughout the six weeks study, yielding a preventive efficacy of 100%. For the spinosad treatment, efficacy was 100% at 24 hours and 48 hours through Day 16, and thereafter declined. The results observed in the spinosad-treated dogs were highly variable between animals. At the 24 and 48 hours counts following the Day 21 infestation, only five of eight spinosad-treated dogs (62.5%) were flea-free. Following the Day  28 infestation, spinosad efficacy fell to 85% and 89%, for the 24 hours and 48 hours counts, and only two dogs (25%) were flea free, compared to 100% flea-free dogs in the fipronil/(S)-methoprene group. No fleas were collected from the fipronil/(S)-methoprene treated dogs throughout the entire study, therefore, no eggs were collected at any time from any dog in the group. However, in the spinosad group adult fleas were found on dogs starting on Day 21 and by Day 30, 42 eggs were collected from one dog that had 107 adult fleas counted at 48 hours. At Day 37 and Day 49, more than 100 eggs were collected from each dog in the spinosad-treated and control groups.

A novel combination of fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) was evaluated for the prevention of attachment of ticks and its ability to cause detachment of ticks. For the two prevention of attachment studies, 20 purpose-bred beagles were allocated each to two equal groups based on pretreatment tick counts (treated and untreated). Each dog was exposed to 50 adult Rhipicephalus sanguineus and Dermacentor variabilis weekly starting 24h after treatment. In study 1 infestations with R. sanguineus were discontinued after Day 7 but continued to Day 28 for D. variabilis in both studies. Counts of ticks by species were made 2, 4 and 24h after exposure to ticks. Ticks not attaching to dogs were evaluated for viability. For the evaluation of detachment study, 16 purpose-bred beagles were allocated each to two equal groups based on pretreatment tick counts (treated and untreated). Each dog was infested with 50 unfed R. sanguineus and D. variabilis adults on Day -2. Ticks were thumb counted without removal on all dogs on Day -1, and at 4, 12, and 24h after treatment. Ticks were counted and removed at 48 h after treatment. Dogs treated with the novel combination had significantly (p<0.05) lower total numbers of attached R. sanguineus and D. variabilis than untreated controls at 4h through Day 7. For R. sanguineus, percent reduction of attachment at 24h after infestation through Day 29 ranged from 94.5% to 100%. For D. variabilis, the percent reduction of attachment at 24h through Day 22 was above 98.0%. These studies demonstrate that novel combination can disrupt attachment of R. sanguineus and D. variabilis for up to 28 days following treatment. Of those ticks that are exposed to the treatment, even if they do not attach to the dog and remain in the environment, greater than 90% (p<0.05) die within 24h for 2-3 weeks following treatment. Also, for those dogs infested with ticks at the time of treatment, the novel combination causes significant detachment (p<.05) starting at 12h and reaching 98.9% by 48 h after treatment. This product provides an effective means for controlling ticks infesting dogs and limiting the spread of tick transmitted diseases. Additionally, the mortality of ticks exposed to CERTIFECT will reduce infestation of the dog's environment., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Four laboratory studies were conducted to demonstrate that a single topical dose of a novel spot-on combination containing fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) is efficacious against the brown dog tick, Rhipicephalus sanguineus. In each study, 6-8 male and 6-8 female purpose-bred, laboratory mongrels, terrier cross or Beagles were randomly assigned to one of two study groups (treated and untreated), based on pre-treatment parasite counts. Starting on the day before treatment, each dog was infested weekly with 50 ticks. Ticks were thumb counted at various time points after treatment and weekly infestations starting as early as 6h and continued at 12, 18 and 24h depending on the study. Ticks were removed and counted at 48 h after treatment and weekly infestations. CERTIFECT provided rapid and excellent control of pre-existing and newly acquired infestations of R. sanguineus with efficacy as high as 93% within the first 12h after a single topical treatment. Excellent control (>96%) of R. sanguineus as early as 18 h, following post treatment infestations was maintained for at least 35 days., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A novel spot-on formulation combining fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) was evaluated in adult Beagle dogs in a study to determine its adulticidal efficacy against the dog flea (Ctenocephalides canis, Curtis, 1826). Sixteen dogs were randomly allocated to treatment groups: 8 dogs served as untreated controls, and 8 dogs were treated once. Treatment consisted of applying a new combination formulation to deliver at least 6.7mg fipronil/kg body weight (bw), 8.0mg amitraz/kg bw, and 6.0mg (S)-methoprene/kg bw. The combination was designed to enhance the efficacy against ticks of the original fipronil/(S)-methoprene combination. Each dog was infested with 100 adult unfed dog fleas within 24h prior to treatment and then at weekly intervals for 8 weeks after treatment. At 24h after treatment or after each subsequent infestation, each dog was combed thoroughly to remove live fleas to be counted. A single treatment with CERTIFECT provided excellent knock-down of fleas within 24h after treatment and controlled re-infestations for up to 7 weeks (efficacy ≥96.5%, p<0.05)., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Four studies were conducted to show the effectiveness of a novel combination of fipronil, amitraz and (S)-methoprene in a spot-on formulation (CERTIFECT™, Merial Limited, GA, USA) for the therapeutic and preventive control of Ixodid tick species affecting dogs in Europe: Ixodes ricinus, Dermacentor reticulatus and Rhipicephalus sanguineus. In each, untreated control dogs were compared to others treated with the novel combination. All dogs were infested with 50 adult, unfed ticks prior to treatment and at 7-day intervals after treatment. Ticks on all dogs were counted at 18, 24 and 48 h after treatment (therapeutic efficacy) or infestation (preventive efficacy). Therapeutic efficacy of fipronil, amitraz and (S)-methoprene was excellent as shown by significant (p<0.05) and greater than 97% and up to 100% reductions in the 48 h tick counts and significant (p<0.05) detachment/death of ticks evident at 18-24h after treatment for all three tick species. Preventive efficacy was demonstrated by significant (p<0.05) and greater than 93% and up to 100% reductions in tick counts at 48 h after repeat infestations out to 35 days after treatment for I. ricinus and out to 42 days after treatment for D. reticulatus and R. sanguineus. The time to substantial disruption of establishment of new tick infestations after treatment was less than 18-24h and was maintained for up to 28 days after treatment of I. ricinus and D. reticulatus infestations, and 4h to at most 18 h and maintained up to 35 days after treatment of R. sanguineus. Similar preventive efficacy profiles for each of the Ixodid species tested suggest that CERTIFECT kills all Ixodid species starting 4h after contact as demonstrated for R. sanguineus., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Five laboratory studies were conducted to confirm that a single topical dose of the novel combination of fipronil, amitraz and (S)-methoprene, CERTIFECT™ (Merial Limited, GA, USA), is efficacious for the rapid control of pre-existing infestations and the prevention of new infestations with Ixodes scapularis, Dermacentor variabilis, Amblyomma americanum and Amblyomma maculatum for at least 28 days on dogs. In each study, 8 male and 8 female purpose-bred, laboratory beagles were randomly assigned to one of two study groups (treated and untreated). Starting on the day before treatment, each dog was infested weekly with about 30 or 50 ticks, depending on the study. Treatment with the novel combination rapidly eliminated pre-existing infestations and controlled weekly re-infestations for at least 28 days. Pre-existing infestations with all four tick species were rapidly and effectively reduced, with post-treatment therapeutic efficacies ranging from 91.7 to 99.5% within 18-48 h post treatment. Amblyomma maculatum numbers were significantly (p<0.05) reduced on treated dogs from the first tick counts as early as 6h post-treatment. All subsequent infestations with each of the 4 tick species were quickly disrupted, with prophylactic efficacies greater than 90% within 18-48 h post-infestation for at least a full month. Because the combination of fipronil, amitraz and (S)-methoprene quickly starts disrupting and killing ixodid ticks within hours of treatment, with similar high levels of efficacy maintained for at least 28 days in these and other studies, the authors conclude that a single topical treatment with CERTIFECT may prevent the transmission of most infectious agents carried by ixodid ticks for at least one month., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Healthy, purpose-bred laboratory beagle dogs that had not been exposed to ticks and were seronegative for Borrelia burgdorferi and Anaplasma phagocytophilum were randomly assigned to four groups of eight dogs each. Control group 1 was not treated. Groups 2, 3 and 4 were treated with a single topical application of a new formulation of fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) at 28, 21 or 14 days prior to tick infestation, respectively. Each dog was infested with 25 female and 25 male field-collected adult Ixodes scapularis ticks that had infection rates of 66% for B. burgdorferi sensu stricto and 23% for A. phagocytophilum, as determined by polymerase chain reaction. Two and five days after tick infestation, control dogs had an average of 9.5 and 13.9 attached adult female ticks, respectively, whilst the 24 treated dogs remained tick-free aside from a single tick on the 2nd day after infestation. Serial serological tests demonstrated that the ticks successfully infected 8/8 control dogs with B. burgdorferi and co-infected 6/8 with A. phagocytophilum. B. burgdorferi infection also was confirmed in most control dogs by culture (6/8) and PCR (7/8) of skin biopsies. In contrast, CERTIFECT protected all 24 treated dogs against infection by both B. burgdorferi and A. phagocytophilum, as demonstrated by their negative serological tests throughout the study and the absence of any positive skin biopsy culture or PCR in these dogs., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Four groups of seven dogs were treated topically with a novel combination of fipronil, amitraz and (S)-methoprene in a spot-on formulation (CERTIFECT™, Merial Limited, GA, USA) on 28, 21, 14 and 7 days prior to tick infestation, respectively and acaricidal efficacy and transmission blocking compared with an untreated control group (seven dogs). All dogs were infested with adult Dermacentor reticulatus ticks harbouring Babesia canis canis. Babesia canis canis was transmitted by D. reticulatus to all seven untreated control dogs, confirmed following demonstration of clinical signs, by the detection of B. canis parasites in thin blood smears and B. canis canis PCR-RLB DNA assay on blood and the development of B. canis canis antibody titres by 14-21 days after tick infestation. The majority of treated dogs remained sero-negative for 42 days after infestation. Therefore, the treatment of dogs with CERTIFECT applied up to 28 days prior to infestation with D. reticulatus harbouring B. canis canis, successfully prevented the development of clinical signs of canine babesiosis., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The acaricidal efficacy against Dermacentor reticulatus in dogs of the commercial topical combinations fipronil/(S)-methoprene (FRONTLINE Combo spot-on dog), imidacloprid/permethrin (Advantix) and metaflumizone/amitraz (ProMeris Duo) was evaluated and compared. Three treatment groups and one untreated control group of six adult Beagle dogs each were randomly formed. Each treatment was administered topically once on Day-0, according to the recommended label dose and instructions for use. All dogs were infested weekly with approximately 50 adult unfed D. reticulatus over a period of seven weeks. Ticks were removed and counted approximately 48 hours after each infestation. The percent reduction in numbers of ticks for fipronil/(S)-methoprene was > or = 97% compared to untreated controls for all seven weekly infestations. The percent reductions for imidacloprid/permethrin and metaflumizone/amitraz were satisfactory initially but fell and stayed below 90 % after three weeks. From the third week onwards, fipronil/(S)-methoprene treated dogs had significantly fewer ticks than imidacloprid/permethrin or metaflumizone/amitraz treated dogs (p < 0.05).

To control West Nile Virus in the greater Toronto area of Ontario, Canada, S-methoprene (Altosid XRbriquets 2.1% AI) is applied each year to storm water catch basins. Because the efficacy of the XRbriquets to reduce adult mosquito populations had not been evaluated locally and was influenced by organic debris in a pilot study, we compared the efficacy of the briquets in 17 sediment and debris-filled catch basins versus 20 catch basins that were vacuumed free of debris. Emergence rates approached 100% in the 5 untreated control catch basins. Emergence rates were significantly lower, and S-methoprene was detected more often and at higher levels, in debris-filled basins versus cleaned catch basins. Overall, 20% of pupae emerged from clean catch basins versus only 3% from debris-filled ones, the difference between treatments becoming significant after 26 days. S-methoprene and total organic carbon concentrations in the catch basins were positively correlated (P < 0.001). We hypothesize that S-methoprene is binding to the organic fraction in the water and sediment in the debris-filled basins, prolonging S-methoprene doses, which are reflected in lower mosquito emergence rates.

This field trial was designed to test the efficacy, in terms of treatment and prevention, of the fipronil 10%+(S)-methoprene 9% combination against immature and adult stages of Rhipicephalus sanguineus in naturally infested dogs, and to assess the effect of a single plain water exposure. Twenty-four dogs of various age, sex, weight and coat length were divided into two homogeneous groups, treated (T) and control (C), and housed into twin outdoor kennels. Trial baseline was designed as day 0, when dogs from group T were treated with a commercial spot-on formulation of fipronil 10%+(S)-methoprene 9%, while subjects from group C were left untreated and served as control. After treatment, tick load for each included dog was estimated, for both adult and immature ticks, using the localization and count over the entire body surface at the following time-points: day 2, to evaluate the speed of kill and at days 7, 14, 21 and 28 to assess the persistence of efficacy. The effect of water exposure on the product efficacy was tested at day 14 of the study, when six dogs, homogenously selected from group T, were soaked through with plain water. The overall tick load in dogs from group C was high throughout the entire study period, ranging from 103.2 (day 28) to 161.3 (day 0), and confirmed the high tick pressure. Speed of kill calculated at 48 h post-treatment was slightly higher for adult ticks (96.2%) than for immature stages (91.6%). Compared to the control, dogs treated with the fipronil+(S)-methoprene maintained a significantly lower mean tick load for both adult and immature stages in the four weeks of observation. Persistence of efficacy against immature stages ranged from 97.1% the first week, 99.6% second week, 99.7 third week and 93.1% in the last week. In the same way, efficacy against adult ticks was constantly high, shifting from 94.5% to 92.5%. Overall efficacy (adults+immatures) was the strongest in the first two weeks (i.e., 96.1% and 96.6%) and lowered in the two remaining weeks (i.e., 94.2% and 93.4%). Results showed that water exposure did not affect the activity and persistence of the tested combination. On the basis of this field trial, the fipronil+(S)-methoprene combination represents a highly efficacious product to control all stages of R. sanguineus ticks on dogs., ((c) 2010 Elsevier B.V. All rights reserved.)

A controlled clinical trial was carried out to assess the effectiveness of pyriprole, metaflumizone combined with amitraz, and fipronil-(S)-methoprene commercial spot-on products in preventing adult female Culex pipiens pipiens from feeding on dogs. Twenty-four beagle dogs were tested for their attractiveness to the mosquitoes and ranked accordingly to produce four groups of equivalent sensitivity to mosquitoes; six were treated with the pyriprole spot-on, six with the metaflumizone combined with amitraz spot-on, six with the fipronil-(S)-methoprene spot-on, and six were left untreated. All the dogs were challenged with 50 unfed adult female Culex in cages for one hour seven days before the treatment, and one and seven days after it. The mosquitoes were checked for blood feeding after one hour and for mortality 24 hours after they had been released into the cages. There was a significant reduction in the number of engorged mosquitoes only with the fipronil-(S)-methoprene product and there were significantly more dead mosquitoes with this product and the pyriprole product, but their effects were too small for them to be recommended for use in a dirofilariosis prevention programme.

Four active ingredients--imidacloprid selamectin, fipronil-(S)-methoprene, and metaflumizone--were tested to assess the speed of flea kill against existing flea infestations and subsequent reinfestations. Thirty flea-infested cats were allocated to four treatment groups and one untreated control group. Flea counts were performed 6, 18, and 48 hours after treatment (day 0) and 2, 4, and 24 hours after weekly flea reinfestations. Cats were also reinfested with fleas after the 6- and 18-hour counts on day 0 and after the 2- and 4-hour counts on subsequent count days. Imidacloprid provided significantly greater flea kill at diverse time points. At the 24-hour counts, all compounds showed expected and similar high efficacies. On study day 34, imidacloprid showed the highest efficacy at 24 hours after reinfestation, with 90.8% flea reduction compared with 55.7% to 67.4% in the other treatment groups. A single topical application of imidacloprid provided a high efficacy in the early elimination of adult fleas, limiting the risk of pathogen transmission and flea allergy dermatitis.

Spot-on formulations of metaflumizone and a combination of fipronil-(S)-methoprene were evaluated in adult cats to determine the duration of 24- and 48-hour efficacy and short-term speed of kill against adult cat fleas, Ctenocephalides felis felis. Speed of kill efficacies (at 12, 18, 24, and 48 hours) were assessed against existing (day -1) infestations and against infestations at day 7, and efficacy was assessed 24 and 48 hours after weekly flea infestations through day 42. Cats treated with fipronil-(S)-methoprene had significantly (P<.01) fewer fleas than those treated with metaflumizone at 12 and 18 hours after treatment (day 0) and on the day 7 infestations. Fipronil-(S)-methoprene-treated cats also had significantly (P<.05) fewer fleas than metaflumizone-treated cats for all 24-hour counts from initial treatment on day 0 through infestation day 42 and for the 48-hour counts on day 28 through day 42.

This study evaluated overall efficacy, percentage of dogs free of live ticks, retention of ticks, and efficacy against retained ticks of fipronil (9.8% w/w) + (S)-methoprene (8.8% w/w) and imidacloprid (8.8% w/w) + permethrin (44% w/w) spot-on topical solutions against Dermacentor variabilis-infested dogs. Tick control was significantly greater (P < .05) on dogs treated with fipronil + (S)-methoprene than on control dogs for the entire 30-day study period. Conversely, the combination product of imidacloprid + permethrin demonstrated activity significantly (P < .05) different from that seen on control dogs only up to day 23. Significantly (P < .05) more dogs treated with fipronil + (S)-methoprene were free of live ticks compared with controls on days 3, 9, and 16. There was never a significant difference (P > .05) between control and imidacloprid + permethrin-treated dogs in numbers of dogs free of live ticks. After the initial 10-minute tick exposure, lower numbers of ticks were retained on the imidacloprid + permethrin-treated dogs than on the fipronil + (S)-methoprene-treated dogs. However, when evaluating the effectiveness of the acaricides on "retained ticks," it appears that while some of the ticks were rapidly knocked down on the imidacloprid + permethrin-treated dogs, efficacy against ticks still retained at 10 minutes was never greater than 50%.

The goal of this study was to assess the insect growth regulator activity of the combination product fipronil-(S)-methoprene under a severe challenge model. Gravid fleas were allowed to feed on untreated donor cats for 48 hours before being transplanted onto untreated control cats and treated cats (treated once on day 0); 24 hours later, adult fleas were collected from all cats and counted to assess the 24-hour kill efficacy against the transplanted fleas, and flea eggs were collected and incubated to assess viability. The process was repeated weekly for 11 weeks. The 24-hour efficacy against transplanted adult fleas in the treated group was about 100% for the first 3 weeks and gradually declined to 93.4% by week 6. Egg production numbers were reduced on the treated cats compared with controls, with geometric mean egg counts on treated cats reduced from 76.9% to 96.3% during the initial 6 weeks of the study. The combination product was 100% ovicidal through day 56 and was still about 98% effective against eggs at the end of the study (day 76).

A study was conducted to evaluate the efficacy of selamectin and fipronil-(S)-methoprene against adult cat fleas (Ctenocephalides felis), flea egg production, and the viability of flea eggs collected from treated cats. Cats were infested with approximately 50 adult fleas 2 days before treatment and weekly thereafter; flea eggs were collected and counted on days 0, 1, 2, and 3 and 48 and 72 hours after each weekly flea infestation. Live fleas were collected approximately 72 hours after treatment or infestation. Compared with fipronil-(S)-methoprene, selamectin provided significantly greater control of adult fleas from days 24 to 31 and significantly greater reduction in egg production from days 16 to 45. For the most part, both products significantly impacted larval and adult emergence for the entire 6-week study, with fipronil-(S)-methoprene providing significantly greater reduction in larval and adult emergence at week 6.

This study evaluated the effectiveness of two topical spot-on formulations -- imidacloprid(8.8% w/w)-permethrin (44.0% w/w) and fipronil (9.8% w/w)-(S)-methoprene (8.8% w/w)--to repel, prevent the attachment of, and kill adult Ixodes scapularis and Amblyomma americanum on dogs. Twelve purpose-bred beagles were distributed into three groups of four dogs each; one group served as untreated controls, and each of the other two groups received one of the test products. Dogs were exposed to 25 adult ticks of each species for 10 minutes on posttreatment days 3, 7, 14, 21, and 28. Unattached or repelled ticks were collected and evaluated for viability, and on-dog tick counts were conducted at 3, 24, and 48 hours after tick exposure. The imidacloprid-permethrin formulation provided significant repellency against I. scapularis for up to 3 weeks after treatment, and both formulations provided good overall control of I. scapularis and A. americanum during the study period.

This study evaluated the effectiveness of two topical spot-on formulations, imidacloprid (8.8% w/w)--permethrin (44.0% w/w) and fipronil (9.8% w/w)--(S)-methoprene (8.8% w/w), to repel, prevent the attachment of, and kill adult Rhipicephalus sanguineus and Dermacentor variabilis on dogs. Twelve purpose-bred beagles were distributed into three groups of four dogs each; one group served as untreated controls and each of the other two groups received one of the test products. Dogs were exposed to 25 adult ticks of each species for 10 minutes on posttreatment days 3, 7, 14, 21, and 28. Nonattached or repelled ticks were collected and evaluated for viability, and on-dog tick counts were conducted at 3, 24, and 48 hours after tick exposure. The imidacloprid-permethrin formulation provided significant repellency of R. sanguineus and D. variabilis for up to 3 and 4 weeks after treatment, respectively; and provided good overall control for R. sanguineus and D. variabilis during the study period. The fipronil--(S)-methoprene formulation provided good overall tick control during the study period.

The red imported fire ant, Solenopsis invicta (Buren), is a major pest in the United States because of its painful sting. Toxic bait has been an important management tool against fire ants, but site registrations prohibit applications of most baits on grazed pastures. Extinguish, containing the insect growth regulator methoprene, was selected for this study because it has a broad site registration that includes grazed pastures. The primary objective of this research was to evaluate the efficacy for control of red imported fire ants by using broadcast applications of methoprene bait at a label rate of 1,121 g/ha versus applications around the perimeter of a target area at the reduced rate of 280 g/ha. Grazed pastures in Lee County, Alabama, and Chambers County, Alabama, were selected for this study, with broadcast treatments, perimeter treatments, and controls replicated three times at each site. All mounds were counted and rated using the USDA population index before applications and then at 8 and 16 wk posttreatment. Perimeter applications did not significantly reduce S. invicta mound abundance, but bait treatments significantly reduced mound abundance at 16 wk posttreatment at site 1 where applications were conducted in early evening. However, broadcast applications were not effective at site 2 where treatments were conducted in early morning with warmer temperatures. Emergence of winged alates was observed at 12 wk posttreatment, followed by a high density of incipient mounds that may have masked the full treatment effect of methoprene applications at site 2. Methoprene bait was effective in reducing abundance of S. invicta only when full label rates were applied.

This study compared the efficacy of two of the most widely veterinary-dispensed topical products for control of ticks on dogs: fipronil-(S)-methoprene and imidacloprid-permethrin. Eighteen healthy beagle dogs of both sexes were divided into three groups of six dogs. Group 1 served as the untreated control. Dogs in group 2 were treated with fipronil 10% + (S)-methoprene 9% w/v applied once on day 0, and those in group 3 were treated with imidacloprid 8.8% and permethrin 44% w/v, applied once on day 0. All dogs were infested with approximately 50 unfed Dermacentor reticulatus ticks on days 1, 7, 14, 21, 28, 35, and 42, and ticks remaining were counted and removed 48 hours after each infestation. The 48-hour efficacy of the fipronil-(S)-methoprene combination remained at 100% at all assessment points through and including day 37, declining to 95.30% on day 44. The 48-hour efficacy of the imidacloprid-permethrin combination peaked at 86.46% on day 9, decreased to 73.37% by the third week, and fell to 63.53% by the end of the study (day 44). Significantly (P < .05) fewer ticks were recovered from dogs treated with fipronil-(S)-methoprene or imidacloprid-permethrin than from untreated control dogs at each infestation. Notably, the dogs treated with fipronil-(S)-methoprene had significantly (P < .05) lower tick counts at every assessment than the dogs treated with imidacloprid-permethrin.

The speed of kill of selamectin, imidacloprid, and fipronil-(S)-methoprene against Ctenocephalides felis infestations on cats for one month following a single treatment was evaluated. Eighty cats were randomly allocated so that there were 20 cats in four different treatment groups. On Days -2, 7, 14, 21, and 28, each cat was infested with 100 adult C. felis from the Kansas 1 flea strain. Following initial application only imidacloprid had caused a significant reduction in adult fleas on treated cats within 6 hours, but by 24 hours all three formulations had killed 96.7% of the fleas. At 7 days post treatment, all three formulations reduced flea populations within 6 and 24 hours by 68.4% and 99.4%, respectively. At 21 and 28 days after treatment, none of the formulations killed significant numbers of fleas as compared to controls within 6 hours of infestation. At 28 days after treatment, selamectin, fipronil-(S)-methoprene, and imidacloprid had killed 99.0%, 86.4%, and 72.6% of the fleas within 48 hours of infestation, respectively. This study demonstrates that the speed of kill of residual flea products on cats decreases throughout the month following application. It also demonstrated that selamectin provided the highest level of residual activity on cats against the Kansas 1 flea strain.

Preventing tick bites is a fundamental step towards reducing the impact of tick-borne protozoal, bacterial and viral diseases (TBDs) in humans and animals. The aim of this study was to evaluate the efficacy of a combination of imidacloprid 10%/permethrin 50% and of fipronil 10%/S-methoprene 12% against ticks in naturally infected dogs and to assess methodological parameters to calculate drug efficacy on tick immature stages. From July to August 2004, 45 privately owned dogs of various sexes, ages, breeds, coat length and habits were enrolled in a trial carried out in an area (radius approximately 50km) in Southern Italy. Three homogeneous groups (both for dog population and tick population) were formed: 15 dogs treated with imidacloprid 10% and permethrin 50% spot-on (group A), 15 dogs treated with fipronil 10% and methoprene 12% spot-on (group B) and 15 untreated dogs (group C). The dogs in each group were then sub-grouped according to their age and weight. Two different treatments were administered (time 0 and +28 days) to groups A and B, and the dogs were checked weekly for tick infestation until day +56 post-treatment (p.t.). Twenty-four areas distributed on the whole body surface were examined for ticks at each follow-up, while only at time 0 and at day +56 p.t., ticks were collected from the dogs and identified. For the immature stages a semi-quantitative method was adopted and the load of immature stages was evaluated and grouped into four classes up to day +56 p.t. when the mean number of immature ticks (MIT) for each infection class was evaluated. All the adult ticks collected were identified as brown dog ticks (Rhipicephalus sanguineus). Immature stages were first compared at day +28 p.t.. The efficacy of both products used in groups A and B on adult ticks was high and generally very similar. Conversely, the efficacy of imidacloprid 10% and permethrin 50% against immatures was higher than that of fipronil 10% and methoprene 12% throughout the observation period with statistically significant differences (p<0.05) at day +28 p.t. (i.e. group A=98.52%, group B=72.40%). On the whole, in analysing the efficacy of both products against adult plus immature ticks, it was found that the combination of imidacloprid 10% and permethrin 50% was more effective than fipronil 10% and methoprene 12%, with the differences being statistically significant at day +28 p.t. (group A=98.43%, group B=77.56%).

The inhibitory activities of fipronil (10% (w/v) solution), (S)-methoprene (9% (w/v) solution), and fipronil/(S)-methoprene (10 and 9% (w/v) solution, respectively) combination against eggs and emerging adult cat fleas (Ctenocephalides felis) and adulticidal activity were tested on experimentally infested dogs. Thirty-two Beagle dogs were selected for this study and eight replicates of four animals were formed based on body weight within sex. One dog in each replicate was randomly allocated to treatment with: (1) untreated control; (2) fipronil 10% (w/v) solution, (3) (S)-methoprene 9% (w/v) solution, and (4) fipronil 10% (w/v) and (S)-methoprene 9% (w/v) combination solution. Treatments were applied once topically on Day 0 at the rate of 0.067 ml/kg. On Days -12, -1, 21, and weekly to Day 84 each dog was infested with approximately 200 fleas and comb counted approximately 24h later, or 2 days (our 48 h) after in the case of Day -1 infestation. On Days -11, 1, 22, and weekly to Day 85 each dog was again infested with approximately 200 fleas. Flea eggs were collected over approximately 24 h beginning 3 days after infestation. Fleas were combed off of the dogs and counted at the end of the egg collection period (approximately 96 h count). One aliquot of up to about 100 eggs, if available, from each animal at each infestation time was incubated for approximately 72 h to determine larval hatch and the other for 35 days to determine the number of adults that developed. The 10% (w/v) fipronil spot-on provided excellent control (>95%) of adult fleas on dogs for 5 weeks. Similarly, the combination spot-on of 10% (w/v) fipronil and 9% (w/v) (S)-methoprene provided excellent control of adult fleas, i.e., >95% for 5 weeks. From week 6 post-treatment onward, the relatively low inhibition of adult flea emergence substantiated the lack of significant ovicidal/larvicidal activity in the fipronil (10%, w/v) treatment group. However, the combination product provided excellent (>90%) ovicidal activity for 8 weeks and high (91.4%) inhibition of adult flea emergence for 12 weeks. In addition, a synergistic effect of the two compounds in combination was demonstrated with fipronil enhancing the ovicidal and inhibition of adult flea emergence activity of (S)-methoprene against cat flea eggs. When all stages of the life cycle of the cat flea are considered, the combination spot-on product provided a high level of total flea control yielding a curative effect against adult fleas and inhibition of flea development stages with little to no potential reinfestation pressure on the animal or in the environment for 12 weeks.

Effectiveness and residual activity tests of granular formulations of 2 insect growth regulators (IGRs), s-methoprene and pyriproxyfen, against laboratory-reared larvae of 5 colonized mosquitoes, Aedes aegypti, Aedes albopictus, Aedes taeniorhynchus, Anopheles quadrimaculatus, and Culex nigripalpus, were conducted in the laboratory and outdoors in plastic tubs. Culex quinquefasciatus was exposed to these two IGRs in the laboratory only. Each IGR formulation was applied at 0.02 and 0.05 ppm active ingredient (Al) against 5 of the 6 mosquito species both in the laboratory and the outdoor evaluations, whereas Cx. quinquefasciatus was exposed to 0.2 and 0.4 ppm AI of s-methoprene, and 0.1 and 0.2 ppm AI of pyriproxyfen in the laboratory. s-Methoprene at 0.02 and 0.05 ppm AI resulted in variable levels (<39-100%) of inhibition of adult emergence in the 5 species monitored for 6 weeks after treatment under both test conditions. Aedes taeniorhynchus was the most susceptible to s-methoprene in terms of initial and residual activity. Culex quinquefasciatus and Ae. albopictus were the most tolerant to s-methopene, with maximum emergence inhibitions amounting to 84% in Cx. quinquefasciatus at 0.4 ppm and 44.3% in Ae. albopictus at 0.05 ppm during the 1st week in the laboratory. Pyriproxyfen at comparable treatment rates to s-methoprene caused very high levels (>80-100% in most cases) of initial and residual emergence inhibitions of the tested species in the laboratory as well as outdoors. In several species, pyriproxyfen induced complete inhibition of adult emergence for several weeks after treatment, even at the lower rate of 0.02 ppm. The World Health Organization has recently recommended the use of pyriproxyfen for the control of some mosquito species at specified rates in certain habitats.

Evidence that amphibian deformities are on the rise has prompted a number of researchers to question a variety of natural and man-made substances, including chemicals such as s-methoprene (trademark Altosid), an insect growth regulator applied to water bodies for mosquito control. Despite conclusions reached by the U.S. Environmental Protection Agency and a considerable body of scientific evidence demonstrating that it is unlikely that s-methoprene or any of its degradation products are causing the high rates of deformities found in Minnesota and other states, concerns persist about potential negative impacts of s-methoprene on nontarget species, specifically anurans. Water analyses in field and laboratory conditions and a comparison of reported Altosid use with reported frog deformities in Minnesota demonstrate that a connection between frog deformities and Altosid use is unlikely. These results indicate that factors other than s-methoprene and its degradation products are contributing to the recent outbreak of frog deformities.

Mosquito larval control by biorational larvicides plays a crucial role in mosquito and mosquito-borne disease management. However, the availability of larvicides that meet the criteria of efficacy, safety, and quality is limited and conventional pesticides are no longer preferred for larval control. Although efforts are made to research new active ingredients (AIs), it is equally important to innovate new formulations based on currently available AIs such as microbial agents and insect growth regulators. Studies were therefore conducted to compare the laboratory activity and semi-field efficacy of OmniPrene® G and Altosid® Pellets with DR-tech, both containing 4.25% S-methoprene, at 2.8 kg/ha and 11.2 kg/ha against the yellow fever mosquito Aedes aegypti (L.) in outdoor microcosms. Both products performed equally in bioassays against the test species with comparable inhibition of emergence activities. In the semi-field study, the lower dose of Altosid Pellets at 2.8 kg/ha, showed lower efficacy than OmniPrene G during the initial 6 weeks; this difference became negligible on week 7, followed by higher efficacy in Altosid Pellets on weeks 8 and 9. More uniform efficacy was observed at the higher dose of 11.2 kg/ha. Equal performance was revealed during weeks 2 to 6, with the OmniPrene G outperforming the Altosid Pellets in week 1, but the opposite during weeks 7 to 9. Mortality patterns were similar in both products, i.e., majority of mortality occurred before emergence, although more incomplete emergence was noted in lower doses, particularly in Altosid Pellets. Overall, newly available OmniPrene G provided comparable activity and efficacy with Altosid Pellets against the test species, with the advantages of fast initial AI release and even coverage, particularly when applied at low doses., (Copyright © 2024 by The American Mosquito Control Association, Inc.)

Background: Spinosad consists of spinosyn A and spinosyn D that are produced by the soil-dwelling actinomycete Saccharopolyspora spinosa. It has been used to control a wide variety of arthropod pests of economic importance. Formulations of spinosad have been used to control larval mosquitoes since approximately 2010. However, the target site- and metabolism-based resistance to this neurotoxin has been reported since 2000 in agricultural pests The current studies aim to further evaluate resistance risk and develop resistance prevention tactics in the southern house mosquito Culex quinquefasciatus., Results: The first comparison group involved selection at lethal concentration, 75% (LC 75 ) of technical spinosad for 30 generations, resulting in baseline resistance ratios (RRs) of 51.1-fold at LC 50 and 45.4-fold at LC 90 in Cx. quinquefasciatus. However, under the same conditions, selection by a combination of spinosad and Bacillus thuringiensis subsp. israelensis (B.t.i.), negated resistance development to spinosad, RRs ranging 1.00-1.75-fold at LC 50 and 0.83-1.76-fold at LC 90 . At the same time, the selected population remained susceptible to the combination throughout the selection process, RRs fluctuating 0.74-1.38-fold at LC 50 and 0.63-1.23-fold at LC 90 ., Conclusion: Combination of spinosad and B.t.i. negates resistance development to spinosad, as opposed to spinosad alone in Cx. quinquefasciatus. Moreover, the species tested does not develop resistance to this combination upon repeated exposures, implying the potential for further developing this combination as a viable product for larval mosquito control. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Mosquito coil is commonly used in many African households for protection against mosquito bites. The coil usually has semi-volatile pyrethroids as an active ingredient, which usually diffuse across open space, and the cloud either kills mosquitoes that are exposed, or mosquitoes can be exposed to sublethal doses of the insecticides. This study was conducted to assess the impact of sublethal doses of mosquito coil on the development of insecticide resistance in Aedes aegypti, a major vector for dengue fever and several other arboviral diseases. A laboratory colony of Ae. aegypti was exposed to sublethal doses of a meperfluthrin-based mosquito coil in a Peet-Grady chamber once per generation for 16 generations. The susceptibility of the exposed colony to a diagnostic dose of the mosquito coil as well as to three other insecticides was determined. Three different kdr mutations and five enzyme activities were evaluated in both the exposed and control colonies. After 16 generations of sublethal exposure to mosquito coils, the full diagnostic dose of the coil caused 68% mortality to the exposed colony compared to 100% mortality in the control colony. Mortality caused by deltamethrin (0.05%) was also significantly lower in the exposed colony. The frequency of 1016I kdr mutation as well as MFO and alpha esterase activities were higher in the exposed colony compared to the control colony. This study provides evidence of the development of pyrethroid resistance in an Ae. aegypti population due to sublethal exposure to mosquito coil for 16 generations. Given the large-scale use of mosquito coils in many African households, its role as a pyrethroid resistance selection source should be taken into consideration when designing resistance management strategies., (© 2024 Royal Entomological Society.)

Background: Insect growth regulators (IGRs) generally are considered to have safer eco-toxicological profiles than the more commonly used neurotoxins and metabolic inhibitors, and are extremely effective against several insect groups, including some invasive ant species. However, use of an IGR product in a large-scale eradication program for a widespread invasive ant (Anoploepis gracilipes; yellow crazy ant) was ineffective. We tested the IGR in question (active ingredient: (S)-methoprene) on A. gracilipes colonies in a laboratory environment to evaluate efficacy., Results: We found that treatment with (S)-methoprene resulted in lower egg production with subsequently decreased numbers of larvae, pupae, and workers over the 135 days of the experiment. None of the treated colonies died, and the number of worker ants in treated colonies was 36% of that seen in control colonies 135 days post-treatment. Treated queen egg production was 39% lower than queens in control colonies, but we saw no effect of treatment on the internal physiology of dissected queens. Treatment had no effect on worker activity levels., Conclusion: Our results show that although (S)-methoprene treatment reduced production of larvae, pupae and workers in treated colonies, the magnitude of reduction was lower than might be expected considering the responses of other species against which this IGR has been tested. Our findings highlight a need for testing species-specific responses to IGR-based insecticides in a controlled environment, before broad-scale field applications that could result in suboptimal management of the target species. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Background: While the clinical features were described recently, the histopathological characterisation of trunk-dominant canine pemphigus foliaceus (PF) is lacking, and whether it differs from classic facial or insecticide-triggered PF is unknown., Hypothesis/objectives: This study describes the histopathological findings of trunk-dominant PF, and compares the results to classic facial and insecticide-triggered PF., Animals: Skin biopsies from 103 dogs with clinically characterised trunk-dominant (n = 33), classic facial (n = 26) and insecticide-triggered PF (n = 44) were included., Materials and Methods: Histological sections, randomised and blinded, were scored for over 50 morphological parameters of pustules, epidermis, dermis, adnexa and crusts. Intact pustule area and width were measured by digital microscopy., Results: In trunk-dominant PF, 77 intact pustules were predominantly subcorneal (0.0019-1.940 mm 2 area, 0.0470-4.2532 mm wide), and contained from one to over 100 acantholytic keratinocytes. Pustules had boat acantholytic cells, corneocytes, perinuclear eosinophilic rings, neutrophil rosettes, acantholytic cell necrosis, rafts, cling-ons and/or eosinophils. Peripustular epidermal spongiosis, necrosis and lymphocyte exocytosis occurred, as did follicular pustules. Mixed dermal inflammation often contained eosinophils. Trunk-dominant PF did not differ from the other PF groups except for few parameters, such as having fewer rafts (p = 0.003). Additional autoimmune inflammatory patterns occurred in all PF groups., Conclusions and Clinical Relevance: Trunk-dominant PF and other canine PF variants are histologically similar, which indicates shared pathomechanisms. The identification of common boat acantholytic cells and corneocyte separation has implications for the mechanisms of acantholysis. The diversity of histopathological and polyautoimmunity features support complicated immune mechanisms. Finally, results indicate that diagnostic biopsies cannot differentiate between these PF variants in dogs., (© 2023 The Authors. Veterinary Dermatology published by John Wiley & Sons Ltd on behalf of ESVD and ACVD.)

Background: Mosquitoes serve as the vector of canine heartworm (Dirofilaria immitis), which represents a significant and persistent threat to canine health. A reduction in the longevity and/or reproductive success of mosquitoes that take a blood meal from fluralaner-treated dogs may consequently reduce the local transmission of heartworm and prevent new infections. A novel secondary effect of an oral formulation of the ectoparasiticide fluralaner (Bravecto ® ) against a laboratory strain of the mosquito Aedes aegypti, a potential major vector of canine heartworm, was investigated in this study., Methods: Six dogs were administered a single dose of fluralaner orally in the form of Bravecto ® Chews (at the labeled fluralaner dose of 25 mg/kg body weight), while six control dogs received no treatment. Mosquitoes were fed on blood that was collected from each dog prior to treatment and weekly for 15 weeks post-treatment to assess the continued effects of fluralaner as its serum level decreased. Mosquito fitness was assessed by three parameters: rate of successful blood-feeding, survival, and egg laying., Results: Successful blood-feeding rate was similar between control and treatment groups. In the fluralaner treatment, mosquito survival was significantly reduced within the first 24 h after blood-feeding, for the first 12 weeks post-treatment of the dogs (efficacy range = 33.2-73.3%). Survival of mosquitoes up until a potentially heartworm-infective timepoint (14 days post-blood-feeding) was significantly reduced in the fluralaner-treated group at several timepoints (1, 2, 5, 11, 12, 13, 14, and 15 weeks post-treatment; efficacy range = 49.4-91.4%), but was less consistently reduced at the other timepoints. Egg laying by mosquitoes was almost completely suppressed for the first 13 weeks following treatment of the dogs with fluralaner (treatment efficacy ≥ 99.8%)., Conclusions: Mosquitoes fed blood from fluralaner-treated dogs experienced a significant reduction in survival and fecundity. These findings support the potential for a reduction in heartworm transmission directly by lethal effects on the vector and indirectly through a reduction of the local vector population when mosquitoes are exposed to animals treated with fluralaner., (© 2023. The Author(s).)