Resumo A atividade carrapaticida in vitro do extrato metanólico (EM) e da fração de alcaloides (FA) de Prosopis juliflora foi investigada, frente ao Rhipicephalus microplus, e relacionada com a inibição da enzima acetilcolinesterase (AChE). A predição in silico das interações de alcaloides dessa fração com a AChE1 de R. microplus (RmAChE1) foi realizada por acoplamento molecular. A atividade carrapaticida foi avaliada, utilizando-se os ensaios de imersão de adultos e larvas. Maior efeito sobre parâmetros reprodutivos de teleóginas foi verificado para a FA, com valor de Concentração Efetiva 50% (CE50) (31.6 mg/mL), quatro vezes menor do que o valor do EM (121 mg/mL). A FA induziu mortalidade de larvas (Concentração Letal de 50% - CL50 = 13,8 mg/mL). A inibição da atividade da AChE de larvas do carrapato foi observada para EM e FA, sendo a FA mais ativa (Concentração Inibitória 50%- CI50 de 0,041mg/mL). As análises químicas da FA permitiram a identificação dos alcaloides prosopina (1), juliprosinina (2) e juliprosopina (3). No ensaio in silico, observou-se que esses alcaloides podem interagir com o sítio ativo da RmAChE1, principalmente por ligações de hidrogênio e interações cátion-pi. Os alcaloides de P. juliflora têm atividade carrapaticida contra R. microplus, atuando através do mecanismo anticolinesterásico.

Abstract We investigated the in vitro acaricide activity of the methanolic extract (ME) and alkaloid-rich fraction (AF) of Prosopis juliflora on Rhipicephalus microplus and correlated this effect with acetylcholinesterase (AChE) inhibition. The acaricide activity was evaluated using adult and larval immersion tests. Also, we studied the possible interaction mechanism of the major alkaloids present in this fraction via molecular docking at the active site of R. microplus AChE1 (RmAChE1). Higher reproductive inhibitory activity of the AF was recorded, with effective concentration (EC50) four times lower than that of the ME (31.6 versus 121 mg/mL). The AF caused mortality of tick larvae, with lethal concentration 50% (LC50) of 13.8 mg/mL. Both ME and AF were seen to have anticholinesterase activity on AChE of R. microplus larvae, while AF was more active with half-maximal inhibitory concentration (IC50) of 0.041 mg/mL. The LC-MS/MS analyses on the AF led to identification of three alkaloids: prosopine (1), juliprosinine (2) and juliprosopine (3). The molecular docking studies revealed that these alkaloids had interactions at the active site of the RmAChE1, mainly relating to hydrogen bonds and cation-pi interactions. We concluded that the alkaloids of P. juliflora showed acaricide activity on R. microplus and acted through an anticholinesterase mechanism.

O carrapato Rhipicephalus(Boophilus) microplusé responsável por grandes perdas na pecuária brasileira, principalmente devido à redução no desempenho produtivo dos animais. A resistência do parasita a diferentes classes de acaricidas tem sido amplamente relatada, bem como os problemas de ocorrência de resíduos desses compostos em produtos de origem animal e no ambiente. Esse estudo teve como objetivo avaliar a eficiência de diversos acaricidas sobre populações de R.(B.) microplus provenientes de propriedades rurais da Região Centro-Norte do Estado da Bahia, Brasil. Teleóginas ingurgitadas foram colhidas em diferentes propriedades da região e submetidas a ensaio in vitro de eficiência de acaricidas, baseado na imersão dos carrapatos em soluções de acaricidas feitas seguindo as recomendações dos fabricantes. Os resultados obtidos indicam níveis variados de eficiência das bases estudadas, com os acaricidas benzofenilureia, lactona macrocíclica e fipronil apresentando os índices mais altos de eficiência, 100%, 100% e 97.34%, respectivamente. Pode-se concluir que, para a região estudada, os acaricidas apresentam níveis diversos de eficiência, e muitos deles possuem eficácia de menos de 95%, como recomendado pela legislação brasileira.

The cattle tick Rhipicephalus(Boophilus) microplus is responsible for major losses in the Brazilian livestock, mainly due to reduction in cattle productive performance. Resistance development to major classes of acaricide widely used nowadays has been extensively reported, as well as the occurrence of residues from these compounds in animal products and the environment. This study aimed to evaluate the efficiency of acaricides onR. (B.)microplus collected from rural properties in the North-Central region of Bahia State. Ticks were collected in several cattle farms in the cited region, and an in vitro acaricide efficiency assay was performed based on the immersion of ticks in acaricide solutions made according to manufacturers' recommendations. The results obtained in the experiments indicated varying degrees of efficiency of the several bases studied, with the products benzofenilurea, macrocyclic lactone and fipronil showing the highest levels of efficiency, 100%, 100% and 97.34%, respectively. It was possible to conclude that, for the region studied, there are different degrees of commercial acaricides efficiency, and many present less than 95% effectiveness, value determined as acceptable by the Brazilian legislation.

Domestic dogs are considered to be the main reservoirs of zoonotic visceral leishmaniasis. In this work, we evaluated a protocol to induce Leishmania infantum/Leishmania chagasi-specific cellular and humoral immune responses in dogs, which consisted of two injections of Leishmania promastigote lysate followed by a subcutaneous inoculation of viable promastigotes. The primary objective was to establish a canine experimental model to provide positive controls for testing immune responses to Leishmania in laboratory conditions. After inoculation of viable promastigotes, specific proliferative responses of peripheral blood mononuclear cells (PBMCs) to either Leishmania lysate or recombinant proteins, the in vitro production of interferon-γ by antigen-stimulated PBMCs and a significant increase in circulating levels of anti-Leishmania antibodies were observed. The immunized dogs also displayed positive delayed-type hypersensitivity reactions to Leishmania crude antigens and to purified recombinant proteins. An important finding that supports the suitability of the dogs as positive controls is that they remained healthy for the entire observation period, i.e., more than seven years after infection. Following the Leishmania antigen lysate injections, the infection of dogs by the subcutaneous route appears to induce a sustained cellular immune response, leading to an asymptomatic infection. This provides a useful model for both the selection of immunogenic Leishmania antigens and for immunobiological studies on their possible immunoprotective activities.

A patogenicidade do fungo Beauveria bassiana (isolado 986) foi avaliada no desenvolvimento da fase parasitária do Anocentor nitens, acompanhando o desenvolvimento de larvas não alimentadas até o estágio adulto. Foram utilizados dois tratamentos e um grupo controle com cinco repetições cada. No tratamento 1 (T1), as orelhas dos bovinos foram banhadas com a suspensão fúngica de 10(8) conídios ml-1; no tratamento 2 (T2), as orelhas foram banhadas com água destilada e espalhante adesivo. No grupo controle (C), as orelhas foram banhadas com água destilada. Após 24 horas dos tratamentos, as orelhas de cada animal foram infestadas com 100mg de larvas. Os parâmetros analisados, em cada tratamento, foram: número e peso de fêmeas recuperadas; períodos de pré-postura, postura, incubação, eclosão e longevidade; percentual de eclosão; peso da massa de ovos e da quenógina; índices de eficiência reprodutiva e nutricional. Verificou-se que, no T1 e T2, o número de fêmeas recuperadas, após 35 dias, foi significantemente menor (P<0,05) que no C. O número de fêmeas recuperadas foi 35, 70 e 117 fêmeas, no T1, T2 e C, respectivamente. As fêmeas oriundas das larvas sobreviventes dos grupos tratados não sofreram ação deletéria do entomopatógeno avaliado.

This study evaluated the pathogenicity of the fungus Beauveria bassiana (Isolate 986) on the development of the parasitic phase of Anocentor nitens by accompanying the development of unfed larvae until the adult stage. Two treated and one control group were used with five repetitions each. In the first treated group (T1), cattle ears were bathed with a fungal suspension of 10(8) conidia ml-1, second treated group (T2), the ears were bathed with distilled water and Tween 80 1% and the control group (C), the ears were bathed with distilled water. One day after treatment, the ears of each animal were infested with 100 mg of larvae. The parameters analyzed in each treatment were: number and weight of females recovered; periods of pre-oviposition, oviposition, incubation, eclosion and longevity; percentage of eggs hatched; weight of the egg mass and of females after oviposition. We found that in the two treated groups the number of females recovered was significantly lower (P<0.05) than in the control group. 35, 70 and 117 females were recovered from the T1, T2 and C, respectively. The females obtained from the surviving larvae of the treated groups did not suffer any deleterious effects from the entomopathogen studied.