Do combinations of fipronil, eugenol and carvacrol have synergistic effects against Rhipicephalus sanguineus?

The tick Rhipicephalus sanguineus is one of the main ectoparasites that affects dogs, causing direct and indirect damage to parasitized animals. Currently, infestation control is mainly carried out by using synthetic acaricidal drugs. However, a decrease in efficacy and an increase in resistance to the main therapeutic protocols against tick infestations have been increasingly reported and confirmed, a factor that has driven research into the potential acaricide activity of natural compounds, including in association with synthetic molecules. The aim of this work was to evaluate whether the combinations of fipronil (FIP) and eugenol (EUG), FIP and carvacrol (CAR), and EUG and CAR would have synergistic effects against immature and unfed adult stages of R. sanguineus through in vitro bioassays. Bioassays were carried out using the larval packet test (FAO 2004) adapted for nymphs and adults. The synergistic activity was explored by combining each solution, based on the estimated LC50, in a 1:1 ratio (FIP: EUG, FIP: CAR and EUG: CAR). CompuSyn software was used to evaluate the various pairwise combinations of FIP, EUG and CAR, checking if there was synergism or antagonism between them. FIP and EUG and FIP and CAR showed combination index (CIn) values above 1.45, indicating antagonism. The synergistic activity between EUG and CAR was verified against all unfed phases of R. sanguineus, since the CIn was below 0.70, a value that indicates synergism. The combination of fipronil with either eugenol or carvacrol presented antagonistic effects against R. sanguineus larvae. On the other hand, carvacrol and eugenol had excellent pharmacological synergism against all tick stages with mortality values in the range of 80 to 100%, including the adult stage, which is less susceptible than immature stages.

Ocimum gratissimum essential oil and eugenol against Ctenocephalides felis felis and Rhipicephalus sanguineus: In vitro activity and residual efficacy of a eugenol-based spray formulation.

Fleas and ticks are among the main ectoparasites that affect pets. The indiscriminate and incorrect use of chemical antiparasitics may be related to increased insect resistance and environmental contamination, requiring prospection for active ingredients that are less harmful to animals, humans and the environment. The use of essential oils and their isolated compounds has been reported as a potential alternative to synthetic antiparasitics, but there is a lack of studies involving the design and development of stable and safe natural products-based formulations. Therefore, the aim of this study was to establish LC50 and LC90 of Ocimum gratissimum essential oil and eugenol on immature stages and adults of Ctenocephalides felis felis and Rhipicephalus sanguineus; and to design and to determine the in vitro efficacy and residual effect of a natural product-based spray formulation for flea and tick control in pets. Bioassays were carried out according to the filter paper impregnation technique for fleas and through the larval packet test for ticks. O. gratissimum essential oil and eugenol presented pulicidal and acaricidal activity in vitro against immature stages and adults of C. felis felis and immature stages of R. sanguineus. The greater potency of eugenol against fleas and ticks led to the choice of eugenol as the active ingredient in the pharmaceutical form developed. The developed eugenol-based sprays presented adequate physical and chemical characteristics and stability, had pulicidal and acaricidal efficacy after 24 h and residual effect against fleas for up to 48 days.

Fluazuron orally administered to guinea pigs: pharmacokinetic and efficacy against Amblyomma sculptum.

BACKGROUND: Brazilian spotted fever (BSF), the most lethal tick-borne disease in the Western Hemisphere, is caused by the bacterium Rickettsia rickettsii and transmitted by the bite of Amblyomma sculptum. Capybaras are considered primary hosts of this tick and amplifier hosts of R. rickettsii, generating new infected lineages of A. sculptum in BSF-endemic areas. To define a possible treatment regimen for controlling the tick A. sculptum in capybaras, the aim of this study was to establish an effective fluazuron (FLU) dose to control A. sculptum larvae in artificially infested guinea pigs. METHODS: In Study I (pharmacokinetic and pharmacodynamic analysis), 24 guinea pigs were divided into four equal groups: control group (CG; untreated) and treated groups receiving FLU administered by gavage in three doses: G1-1 mg/kg, G2-5 mg/kg and G3-10 mg/kg, once a day for 15 days (d0 to d + 14). Blood samples were collected from the animals of the treated groups before and at d + 1, + 2, + 4, + 7, + 15 and + 21. The guinea pigs were artificially infested at d + 7 with A. sculptum larvae, and specimens were recovered at d + 11 to d + 14 and kept in a climatized chamber for 14 days. In Study II (evaluation of pharmacokinetic parameters), one group of eight animals received FLU administered by gavage in a single dose of 10 mg/kg, and blood samples were collected before and on day 0 (8 h after treatment), + 1, + 4, + 7, + 15, + 21 and + 28 after single FLU administration. FLU was analyzed in plasma samples by high-performance liquid chromatography with ultraviolet detection. RESULTS: FLU plasma concentrations increased quickly, indicating rapid absorption, and decreased slowly. Some larvae from all treated groups exhibited morphological and behavioral changes. FLU interfered in molting, and the efficacy obtained was 100% for all treated groups. CONCLUSIONS: The results offer promising perspectives for the development of a palatable feed cube containing FLU for free-living capybaras to control A. sculptum and also to prevent BSF in areas where capybaras have been shown to play a primary role.

Oral pharmacokinetic profile of fipronil and efficacy against flea and tick in dogs.

Fipronil (FIP) is an ectoparasiticide of the phenylpyrazole class, used in veterinary medicine in topical form. Supported by evidence of uncontrolled human exposure to FIP and environmental damage caused by commercially available formulations, its use by oral administration has become promising. The effectiveness of FIP against the flea Ctenocephalides felis felis and the tick Rhipicephalus sanguineus and its pharmacokinetics and main active metabolite, fipronil sulfone (SULF) were evaluated after single oral administration of tablets in three different doses (2, 4, and 6 mg/kg) in dogs. Through the plasma concentration curves, it was possible to observe that the FIP showed rapid absorption and metabolization and slow elimination. The values of Cmax (ß = 0.7653) and AUC0- t (ß = 0.3209) did not increase proportionally with increasing dose. At 48 h after treatment, doses of 4 mg/kg (AUC0- t  = 442.39 ± 137.35 µg/ml*h) and 6 mg/kg (AUC0- t  = 421.32 ± 102.84 µg/ml*h) provided 100% and 99% efficacy against fleas, and 95% and 98% against ticks, respectively. The estimated EC90 of FIP +SULF was 1.30 µg/ml against C. felis felis and 2.16 µg/ml against R. sanguineus. The correlation between the FIP pharmacokinetic and efficacy data demonstrated its potential for oral administration in the form of tablets for the control of ectoparasites in dogs, as a safer alternative for animals, humans, and the environment, aligned with the One Health concept.

Efficacy and residual effect of Illicium verum (star anise) and Pelargonium graveolens (rose geranium) essential oil on cat fleas Ctenocephalides felis felis.

The essential oils (EOs) of Illicium verum and Pelargonium graveolens were evaluated for lethality, inhibition of development and residual efficacy against the flea Ctenocephalides felis felis. Their chemical composition was characterized by means of gas chromatography with a flame ionization and mass spectrometry detection. Mortality at different immature stages and among adult fleas was measured through in vitro filter paper tests at different concentrations of EOs. The chemical characterization of I. verum volatile oil showed that E-anethole (79.96%) was the major constituent, while the major compounds in P. graveolens were citronellol (29.67%) and geraniol (14.85%). Insecticidal activity against both immature and adult flea stages were observed. The EO of I. verum had insecticidal activity for approximately 18 days, while the EO activity of P. graveolens lasted for 13 days. The pulicidal activity of I. verum remained above 70% for up to 9 days, while the activity of P. graveolens was 41.7% for up to 2 days. Essential oils, especially that of I. verum, showed insecticidal activity for flea control at different life cycle stages and have potential for the development of ectoparasiticides (biopesticides) for veterinary use.

Evaluation of pyriproxyfen in cattle by oral treatment: An alternative to control Haematobia irritans.

The horn fly, Haematobia irritans, is a bovine ectoparasite that causes large losses to cattle breeders, through lower meat and milk production and hide damage. Currently, the control of this parasite is mainly through chemicals. However, the indiscriminate use of these substances generates resistance. Pyriproxyfen belongs to an insect growth disruptors class with mechanisms of action for the control of immature forms of the insect. The aim of this study was to investigate the potential of pyriproxyfen administered orally to cattle for the control of the horn fly. In vitro bioassays were carried out by evaluating the number of adults emerged from 30 eggs per replicate, in triplicate, added in a substrate (fresh cattle feces) spiked with pyriproxyfen solutions in the concentration range of 2-130 ppb (ng.g-1). Probit analysis estimated LC50 value of 7.89 ppb and LC90 value of 70.08 ppb. The doses used in the in vivo evaluation were established based on the LC90 values and the anatomical and physiological aspects of the bovine digestive tract. Capsules containing pyriproxyfen at doses of 2.5 mg (G2.5) and 40 mg (G40) were produced and administered orally by bolus applicator for 12 days. The efficacy of pyriproxyfen against the immature forms of H. irritans was determined by incubating eggs in vitro in the feces collected on days 0, +3, +6, +10 and +13. Quantification of pyriproxyfen in feces was performed by UPLC-MS/MS, finding concentrations ranging from 13.4 to 22.6 ppb for G2.5 and between 268.5 to 509.0 ppb for G40. Pyriproxyfen administered orally is eliminated in the active form in the fecal mass and at a dose of 40 mg.day-1 (0.1 mg/kg/day) generates fecal concentrations able to produce 100 % prevention of adults emergence of H. irritans.

Efficacy and residual effect of Illicium verum (star anise) and Pelargonium graveolens (rose geranium) essential oil on cat fleas Ctenocephalides felis felis / Eficácia e efeito residual dos óleos essenciais de Illicium verum (anis-estrelado) e Pelargonium graveolens (gerânio rosa) em pulgas de gato Ctenocephalides felis felis

Abstract The essential oils (EOs) of Illicium verum and Pelargonium graveolens were evaluated for lethality, inhibition of development and residual efficacy against the flea Ctenocephalides felis felis. Their chemical composition was characterized by means of gas chromatography with a flame ionization and mass spectrometry detection. Mortality at different immature stages and among adult fleas was measured through in vitro filter paper tests at different concentrations of EOs. The chemical characterization of I. verum volatile oil showed that E-anethole (79.96%) was the major constituent, while the major compounds in P. graveolens were citronellol (29.67%) and geraniol (14.85%). Insecticidal activity against both immature and adult flea stages were observed. The EO of I. verum had insecticidal activity for approximately 18 days, while the EO activity of P. graveolens lasted for 13 days. The pulicidal activity of I. verum remained above 70% for up to 9 days, while the activity of P. graveolens was 41.7% for up to 2 days. Essential oils, especially that of I. verum, showed insecticidal activity for flea control at different life cycle stages and have potential for the development of ectoparasiticides (biopesticides) for veterinary use.

Resumo Os óleos essenciais (OE) de Illicium verum e Pelargonium graveolens foram avaliados quanto à letalidade, inibição do desenvolvimento e eficácia residual contra a pulga Ctenocephalides felis felis. Sua composição química foi caracterizada por meio de cromatografia gasosa com detector de ionização de chama e espectrometria de massas. A mortalidade entre os diferentes estágios imaturos e pulgas adultas foi avaliada por meio de testes in vitro em papel filtro, contendo diferentes concentrações de OEs. A caracterização química do óleo volátil de I. verum mostrou que o E-anetol (79,96%) foi o constituinte majoritário, enquanto os principais compostos de P. graveolens foram citronelol (29,67%) e geraniol (14,85%). Foi observada atividade inseticida contra os estágios imaturos e adulto da pulga. O OE de I. verum teve atividade inseticida por aproximadamente 18 dias, enquanto o de P. graveolens durou 13 dias. A atividade pulicida de I. verum permaneceu acima de 70% até o 9º dia, enquanto a atividade de P. graveolens foi de 41,7% até o 2º dia. Os óleos essenciais, principalmente de I. verum, apresentam atividade inseticida para o controle de pulgas em diferentes estágios do ciclo de vida e têm potencial para o desenvolvimento de ectoparasiticidas (biopesticidas) de uso veterinário.

In vitro acaricidal activity of different ectoparasiticide classes against Amblyomma sculptum larvae.

Zoonoses are major causes of morbidity and mortality worldwide. Among them, Brazilian Spotted Fever (BSF) is an important one that occurs in some regions of South America and can be transmitted by the "star tick" Amblyomma sculptum. Application of acaricides against the larval stage is important as strategy of population control. However, there is still a deficiency of studies on chemical control of A. sculptum and the present work aims to evaluate the in vitro acaricidal activity of cypermethrin, flumethrin, deltamethrin, fipronil, coumaphos and chlorpyrifos against A. sculptum larvae. Bioassays were performed using the larval immersion test method. A discriminatory analysis between the antiparasitic classes most used for tick control was carried out, which made it possible to determine the classes with higher potential for controlling A. sculptum larvae. Our results showed that A. sculptum larvae present highest sensitivity to the synthetic pyrethroid group, followed by the phenylpyrazole, organophosphate and macrocyclic lactone groups. These findings may support studies on improvement of tick control as in animals as in the environment.

Fipronil Tablets: Development and Pharmacokinetic Profile in Beagle Dogs.

Increased human-pet interactions have led to concerns related to the prevention and treatment of ectoparasite infestations. Fipronil (FIP) is a widely used ectoparasiticide in veterinary medicine available for topical administration; however, its use may cause damage to the owners and the environment. The aim of the study was to develop immediate-release tablets of FIP, as well as to determine its pharmacokinetic properties after oral administration in beagle dogs. The prepared FIP tablets were evaluated for pre-compression (angle of repose, speed flow, and Carr's index) and post-compression (weight variation, friability, thickness, hardness, disintegration time, and dissolution rate) parameters. Orally administered FIP at a dose of 2 mg/kg was rapidly absorbed with Cmáx of 3.13 ± 1.39 µg/mL at 1.83 ± 0.40 h post treatment (P.T.) and metabolized with 1.27 ± 1.04 µg/mL at 2.33 ± 0.82 h P.T. for fipronil sulfone (SULF) (the primary metabolite). The elimination of FIP and SULF occurred slowly and had maintained quantifiable plasma levels in the blood for up to 28 days P.T. The goal of the study is aligned with the concept of One Health, which aims to collaboratively achieve the best health for people, animals, and the environment. Therefore, the use of FIP tablets for the control of ectoparasites in dogs may be a safer alternative for owners and the environment.