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.

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.

Pyothorax caused by Nocardia sp. in a cat - Case report.

Nocardiosis is an infection caused by ubiquitous opportunistic bacteria and is rare in felines. In retrospective studies of the infection in the species, pyothorax was found in only about 5% of the cases described, with the most frequent forms being cutaneous and subcutaneous. This report describes Nocardia spp. as an etiologic agent of pyothorax in a feline leukemia virus (FeLV)-positive female cat. A 10-year-old female mixed-breed cat, positive for FeLV, with chronic kidney disease IRIS stage 2, was examined with a severe dyspneic onset, suspected of pleural effusion. It also had skin lesions in the interscapular region. The patient underwent thoracentesis, and a dense, creamy, fetid, and flocculated liquid was collected. Radiography was performed after draining the pleural fluid, which showed images suggestive of atelectasis of the left lung lobes and pneumonia. The collected material was sent for analysis, which found that it was a septic exudate. In addition, it was referred for bacterial culture in a special medium due to suspicion of nocardiosis. Long-term treatment with amoxicillin + clavulanic acid resolved the clinical case.

A nocardiose é uma infecção causada por bactérias oportunistas ubíquas e é rara em gatos. Em estudos retrospectivos da infecção na espécie, o piotórax foi encontrado em apenas cerca de 5% dos casos descritos, sendo as formas mais frequentes a cutânea e a subcutânea. Este relato descreve Nocardia spp. como agente etiológico do piotórax em um gato vírus da leucemia felina (FeLV)-positivo. Um gato mestiço de 10 anos de idade, positivo para o FeLV, com doença renal crônica IRIS estágio 2, foi examinado com dispneia grave, com suspeita de derrame pleural. Apresentava lesões cutâneas na região dorsal entre as escápulas. Após sedação, tricotomia e antissepsia, o paciente foi submetido à toracocentese e coletado líquido denso, cremoso, fétido e floculado. A radiografia foi realizada após drenagem do líquido pleural, que mostrou imagens sugestivas de atelectasia dos lobos pulmonares esquerdos e pneumonia. O material coletado foi encaminhado para análise, que constatou tratar-se de exsudato séptico. Além disso, foi encaminhado para cultura bacteriana em meio especial por suspeita de nocardiose. O tratamento prolongado com amoxicilina + ácido clavulânico foi suficiente para a resolução do caso clínico.

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.