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.

Evaluation of the susceptibility to fipronil of Rhipicephalus microplus larvae from egg masses incubated at different times of oviposition.

The objective of this work was to evaluate the susceptibility of R. microplus larvae from different oviposition times to fipronil. The LPT was performed in sextuplicate, at concentrations of 18.75, 37.5, 75, 150 and 300 µg.mL-1. The LC50 found for the egg masses incubated with +7, +14 and +21 days were respectively 105.87, 110.71 and 121.22 µg.mL-1. The larvae originating from egg masses from the same group of engorged females, incubated on different days, presented similar mortality rates compared to the evaluated fipronil concentrations, facilitating the maintenance of laboratory colonies of this tick species.

O objetivo deste trabalho foi avaliar a susceptibilidade de larvas de R. microplus oriundas de diferentes momentos da oviposição frente ao fripronil. O TPL foi realizado em sextuplicata, nas seguintes concentrações 18,75; 37,5; 75; 150; 300 µg.mL-1. Não houve diferença estatística entre as mortalidades das larvas oriundas de posturas incubadas nos dias mais sete, +14 e +21, expostas ao fipronil. As CL50 encontradas para as posturas incubadas com +7, +14 e +21 dias foram respectivamente 105,87; 110,71 e 121,22 µg.mL-1. Observou-se que as larvas oriundas de posturas, do mesmo grupo de fêmeas ingurgitadas, incubadas em dias diferentes apresentam taxas de mortalidade parecidas frente as concentrações de fipronil avaliadas, facilitando a manutenção das colônias laboratorias desta espécie de carrapato.

Insecticidal activity in vitro of the essential oil of Pogostemon cablin against Ctenocephalides felis felis.

The aim of this study was to evaluate the in vitro insecticidal activity of the essential oil (EO) of P. cablin against immature and adult stages of C. felis. The chemical composition of P. cablin EO was determined by gas chromatography (GC). To evaluate the insecticidal activity, the egg, larva, pupa, and adult stages of C. felis were exposed to different concentrations of P. cablin EO diluted in acetone and impregnated in filter paper in a concentration range of 25 to 200 µg.cm-2. After exposure, the material was incubated in climatized chambers and mortality assessment was performed after 24 h for the larval and adult stages, 72 h for the egg stage and 15 days for the pupal stage. After GC analysis, the major constituents of the EO of P. cablin were found to be patchoulol (27.4%), α-bulnesene (19.7%) and α-guayene (15.0%). The of mortality percentage was 100% at the concentration of 200 µg.cm-2 for all stages. The LCs50 values were 46.1, 49.2, 60.6 and 62.7 µg.cm-2, respectively for the pupal, egg, larva and adult stages. Based on the results obtained, we can conclude that the EO of P. cablin contained patchoulol, α-bulnesene, and α-guayene as major constituents and had insecticidal activity against the immature and adult stages of C. felis.

O objetivo deste estudo foi avaliar a atividade inseticida in vitro do óleo essencial (OE) de Pogostemon cablin frente estágios imaturos e adultos de Ctenocephalides felis. A composição química do OE de P. cablin foi determinada por cromatografia gasosa (CG). Para a avaliação da atividade inseticida os estágios de ovo, larva, pupa e adultos de C. felis foram expostos a diferentes concentrações do OE de P. cablin diluídos em acetona e impregnados em papel filtro com a faixa de concentração de 25 a 200 µg.cm-2. Após a exposição, o material foi incubado em câmaras climatizadas e a avaliação da mortalidade foi realizada após 24 h para os estágios de larvas e adultos, 72 h para o estágio de ovo e 15 dias para o estágio de pupa. Após análise cromatográfica, os constituintes majoritários para o OE de P. cablin foram: patchoulol (27,4%), α-bulneseno (19,7%) e α-guaieno (15%). Foi obtido 100% de mortalidade na concentração de 200 µg.cm-2 para todos os estágios. As CLs50 foram de 46,1; 49,2; 60,6 e 62,7 µg.cm-2, respectivamente para os estágios de pupa, ovo, larva e adultos. Com base nos resultados obtidos é possível concluir que o OE de P. cablin apresentou o patchoulol, α-bulneseno, e α-guaieno como constituintes majoritários e atividade inseticida frente aos estágios imaturos e adultos de C. felis.

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.