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.

In vitro evaluation of the activity of Cinnamaldehyde as an inhibitor of the biological cycle of Ctenocephalides felis felis.

The objective of this work was to evaluate the in vitro activity of cinnamaldehyde as an inhibitor of the biological cycle of Ctenocephalides felis felis. Tests were carried out with six repetitions containing 10 eggs of C. felis felis together with 0.5g of diet and filter paper discs with area of 23.76 cm2 impregnated with different concentrations of cinnamaldehyde (6; 8; 10; 12; 14; 16; 18; 20; 24 and 28 µg.cm-2) in Petri dishes and incubated in a climate-controlled chamber at 27 ± 1 °C and RH 75 ± 10%, for 30 days. The tests included a placebo group containing the diluent (acetone) and a negative control group, without treatment. At the end of 30 days, the percentage of inhibition of adult emergence/mortality was calculated, as well as the LC50 and LC90. Inhibition of adult emergence greater than 50% was observed from a concentration of 10 µg.cm-2 and 100% inhibition from a concentration of 14 µg.cm-2, obtaining LC50 and LC90 results of 8.75 and 13.57 µg.cm-2, respectively. We concluded that the volatile compound cinnamaldehyde is effective in vitro as an inhibitor of the biological cycle of C. felis felis.

O objetivo deste trabalho foi avaliar a atividade in vitro do cinamaldeído como inibidor do ciclo biológico de Ctenocephalides felis felis. Para realização do teste, seis repetições contendo 10 ovos de C. felis felis juntamente 0,5g de dieta e papel filtro de área 23,76 cm2 impregnado com diferentes concentrações de cinamaldeído (6; 8; 10; 12; 14; 16; 18; 20; 24 e 28 µg/cm2) foram armazenados em placas de petri e incubados em câmara climatizada à 27 ± 1°C e UR 75 ± 10%, por 30 dias. Além disso, o teste contou com um grupo placebo contendo o diluente (acetona) e um grupo controle negativo, sem tratamento. Ao final dos 30 dias, com os dados coletados foram calculados o percentual de inibição de emergência de adultos/mortalidade, como também a CL50 e CL90. Com os resultados, foi observado uma inibição de emergência de adultos superior a 50% a partir da concentração de 10 µg.cm-2 e uma inibição de 100% a partir da concentração 14 µg.cm-2 obtendo como resultados de CL50 e CL90 de 8,75 e 13,57 µg.cm-2, respectivamente. Conclui-se que o composto volátil cinamaldeído tem eficácia in vitro como inibidor do ciclo biológico de C. felis felis.

Prevalence of gastrointestinal parasites in domestic cats (Felis catus) diagnosed by different coproparasitological techniques in the municipality of Seropédica, Rio de Janeiro.

The objectives of this work were to investigate the occurrence of parasites in feces of cats, compare different coproparasitological techniques for their diagnosis and determine associations with parasitism. The samples were processed using three different coproparasitological techniques: centrifugal flotation in sucrose, centrifugal flotation in ZnSO4 and simple sedimentation. The parasitic association between parasitism and variables such as age, sex and fecal consistency was performed using the chi-square test or the G test with a significance level of 5%. A total of 237 samples were analyzed, of which 93 (39.2%) were positive, being Ancylostoma spp. (17.3%), Giardia intestinalis (12.2%), Platynosomum illiciens (8.0%), Cystoisospora spp. (6.3%), Toxoplasma gondii/ Hammondia hammondi (3.4%), Diphyllobothriidae (2.1%), Toxocara spp. (1.7%), Dipylidium caninum (1.3%) and Mesocestoides spp. (0.8%). In the parasitism association analysis, it was possible to verify a statistical difference in the age category for Cystoisospora spp. (p=0.001) observing a strong relationship between parasitism and young animals, the association with sex proved to be important for P. illiciens (p<0.001) with a higher frequency of parasitized females and fecal consistency revealed to be related to the parasites G. intestinalis (p=0.007) and P. illiciens (p=0.033) showing a higher number of positive animals for these parasites with normal fecal consistency. In conclusion, we observed a higher occurrence of Ancylostoma spp. and G. intestinalis in fecal samples from domestic cats received in routine diagnoses and the presence of other parasites with zoonotic potential, as well as the relationship of these diagnosed parasites with the categories sex, age and fecal consistency.

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.

Endoparasites of Leopardus pardalis (Carnivora, Felidae) rescued in the State of Rio de Janeiro.

Wild cats play an important role as top predators in the food chain and act as ecosystem regulators. However, in recent decades, many studies have demonstrated the potential effects of parasitic diseases on wild carnivore populations, including cats. This study reports on the endoparasites found in an injured and rescued specimen of Leopardus pardalis in the state of Rio de Janeiro. Fecal samples were collected and processed using five coproparasitological techniques: a simple flotation centrifugation, a zinc sulfate flotation centrifugation, a formalin-ether sedimentation centrifugation, a conical centrifuge tube technique, and a modified Ziehl-Neelsen staining technique for fecal smears. Helminth eggs belonging to the families Trichuridae and Diphyllobothriidae and the genus Toxocara were found in both sedimentation flotation techniques. Protozoan oocysts belonging to the genus Cryptosporidium were identified by modified Ziehl-Neelsen staining. These findings show that ocelots can harbor potentially zoonotic and pathogenic endoparasites. Further studies on the helminths and protofauna of these animals are necessary.

Os felinos selvagens desempenham um papel importante como predadores de topo na cadeia alimentar e atuam como reguladores do ecossistema. No entanto, nas últimas décadas, muitos estudos demonstraram os efeitos potenciais de doenças parasitárias em populações de carnívoros selvagens, incluindo gatos. Este estudo relata os endoparasitos encontrados em um espécime ferido e resgatado de Leopardus pardalis no estado do Rio de Janeiro. Amostras fecais foram coletadas e processadas usando cinco técnicas coproparasitológicas: uma centrifugação por flotação simples, uma centrifugação por flotação com sulfato de zinco, uma centrifugação por sedimentação com éter de formalina, uma técnica de tubo de centrífuga cônica e uma técnica modificada de coloração de Ziehl-Neelsen para esfregaços fecais. Ovos de helmintos pertencentes às famílias Trichuridae e Diphyllobothriidae e ao gênero Toxocara foram encontrados em ambas as técnicas de flotação por sedimentação. Oocistos de protozoários pertencentes ao gênero Cryptosporidium foram identificados pela coloração de Ziehl-Neelsen modificada. Esses achados mostram que jaguatiricas podem abrigar endoparasitas potencialmente zoonóticos e patogênicos. Mais estudos sobre os helmintos e a protofauna desses animais são necessários.