Plants used in Côte d'Ivoire (West Africa) against ticks: Evaluation for acaricidal activity against Rhipicephalus (Boophilus) microplus.

Rhipicephalus (Boophilus) microplus is a tick that is now a real constraint to cattle breeding in many West African countries such as Côte d'Ivoire. Antitick control is difficult because of the cost, ecotoxicity and resistance. The main objective of this study was to evaluate the acaricidal activity of plants used in Côte d'Ivoire for tick infestation control in cattle. Ethanol extracts from 17 plants were tested in vitro on eggs, larvae and adults of two amitraz resistant strains of R. (B.) microplus, using respectively LTT (Larval Tarsal Test), LPT (Larval Packet Test) and AIT (Adult Immersion Test) methods, at concentrations of 5 mg/mL -1 mg/mL. Out of the 22 extracts tested, four showed acaricidal activity against eggs (4 mg/mL) and three against larvae at 5 mg/mL. These extracts inhibited fertility and resulted in adult females mortality. These results give credence to the traditional use of active plants that may be used for plant-based bioacaricide development and R. (B.) microplus control in cattle as well.

Invitro acaricidal activity of ethnoveterinary plants and green synthesis of zinc oxide nanoparticles against Rhipicephalus (Boophilus) microplus.

The present study was designed to investigate the invitro acaricidal effects of seven ethnoveterinary plants, zinc acetate and green synthesized zinc oxide nanoparticles against the Southern cattle tick, Rhipicephalus (Boophilus) microplus. The selected ethnoveterinary plants were extracted using ethanol and aqueous (water) solvents at 0.02mg/ml and 0.04mg/ml concentrations. Of these seven plants, Lobelia leschenaultiana showed the highest percentage of tick mortality. The ethanol extracts of L. leschenaultiana showed 93.33% mortality at 0.04mg/ml and its LC50 was 0.05mg/ml. However, zinc acetate exhibited 70% mortality at 0.04mg/ml (LC50: 0.0192mg/ml). Further, we synthesized ZnO nanoparticle using the leaf extracts of L. leschenaultiana and zinc acetate as the precursor material to control R. (B.) microplus. The structural characterization of the synthesized ZnO nanoparticles (Ll-ZnO NPs) was performed by UV-vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM) and transmission electron microsopy (TEM). UV-vis spectra showed the absorption band at 383nm. XRD analysis clearly showed the crystalline nature of Ll-ZnO NPs with various Bragg's reflection peaks at 100, 002, 101, 102, 110, 103, 200, 201 and 202 planes. FTIR analysis showed the possible functional groups of Ll-ZnO NPs with strong band at 3420.63 and 2922.48cm(-1). SEM and TEM analysis revealed that the Ll-ZnO NPs were spherical and hexagonal in shape with particle size ranging between 20 and 65nm. The mortality of R. (B.) microplus after treatment with Ll-ZnO NPs was 35, 57.5 and 82.5% at 0.001, 0.002 and 0.004mg/ml. On the otherhand, 100% mortality of R. (B.) microplus was observed at 0.008mg/ml (LC50: 0.0017mg/ml). The results indicated that the Ll-ZnO NPs have good acaricidal properties compared to L. leschenaultiana leaf extract and zinc acetate.

High efficacy of a 20 amino acid peptide of the acidic ribosomal protein P0 against the cattle tick, Rhipicephalus microplus.

Current strategies to control cattle ticks use integrated control programs (ICP) that include vaccination. Reduction in the use of chemicals and in the cost of tick control, the delay or elimination of acaricide resistance and the decreasing of environmental pollution are the advantages of using these programs. This integrated program is potentially applicable to all genotypes of chemical resistant ticks. However, the problem here is to improve the efficacy of anti-tick vaccines. The P0 protein is a structural component of the ribosome of all organisms. We have identified an immunogenic region of ribosomal protein P0 from Rhipicephalus spp. ticks that is not very conserved compared to the orthologous protein in their hosts. A synthetic 20 amino acid peptide from this sequence was effective as a vaccine against Rhipicephalus sanguineus infestations in an immunization and challenge experiment using rabbits. In this paper, the same peptide used as vaccine against the cattle tick Rhipicephalus Boophilus microplus shows a significant diminution in the number of engorged females recovered, in the weight of females and the weight of egg masses. The number of eggs hatched was also significantly reduced for the vaccinated group, with an overall effectivity for the antigen pP0 of 96%. These results, together with the conserved sequence of the P0 peptide among ticks, suggest that this antigen could be a good broad spectrum vaccine candidate. It would be expected to be active against many species of ticks and thus has promise in an ICP for effective control of ticks and thereby to improve the efficiency and productivity of the livestock industry.

Effect of a spray formulation on the reproductive parameters of a susceptible population of Rhipicephalus (Boophilus) microplus / Efeitos de uma formulação pulverização sobre os parâmetros reproductivos de uma população susceptível de Rhipicephalus (Boophilus) microplus

The number of studies emphasizing the possible damage that acaricidal spray formulations can cause on engorged female ticks' reproductive parameters is small. The present study evaluated the deleterious effects of a spray formulation (dichlorvos 60% + chlorpyrifos 20%) on the reproductive parameters of a susceptible population of Rhipicephalus (B.) microplus females, using the Stall Test. The ticks were allocated randomly to treatments according to the mean numbers of females detached from each cow on days -3, -2 and -1 and the cattle pen location. The numbers of engorged female ticks that naturally detached from the cattle were counted daily from day 1 to day 30. For each group, 20 detached engorged female ticks or the available number collected daily were evaluated regarding reproductive parameters. Associations of organophosphates demonstrated elevated acaricidal efficacy, as well as deleterious effects on the reproductive parameters of R. (B.) microplus females. The engorged female weight (days 1 to 7), weight of egg masses (days 5 to 10) and larval hatching percentage (days 5 to 19) were decreased (P ≤ 0.05). It is possible that a formulation can lead to deleterious effects on R. (B.) microplus females when the tick population analyzed shows elevated sensitivity towards a particular formulation. However, further studies need to be conducted.

É relativamente pequeno o número de estudos que enfatiza os danos que uma formulação acaricida spray pode desencadear sobre os parâmetros reprodutivos das teleóginas. O presente estudo teve como objetivo avaliar os efeitos deletérios de uma formulação spray comercial (dichlorvos 60% + Clorpirifós 20%), sobre os parâmetros reprodutivos de uma população susceptível de R. (B.) microplus, desprendidas de bovinos experimentalmente infestados, utilizandose o teste de estábulo. Os animais foram alocados aos grupos de tratamentos de acordo com a contagem média de fêmeas desprendidas dos bovinos nos dias -3, -2 e -1. O número de teleóginas desprendidas foi quantificado do dia 1 ao 30. Para cada grupo, diariamente 20 fêmeas, ou a quantidade disponível, foram selecionadas e submetidas à avaliação dos parâmetros reprodutivos. A associação de organofosforados demonstrou elevada eficácia acaricida e também apresentou efeitos deletérios sob os parâmetros reprodutivos de Rhipicephalus (B.) microplus, diminuindo (P≤0,05) o peso das teleóginas (dos dias 1 ao 7), o peso da massa de ovos (dos dias 5 ao 10) e a eclodibilidade das larvas (dos dias 5 ao 19). Talvez uma formulação pode apresentar efeitos deletérios sobre os parâmetros reprodutivos de fêmeas de R. (B.) microplus, quando existe um elevado grau de sensibilidade dessa cepa de carrapato a um determinado composto. De qualquer maneira, futuros estudos devem ser realizados.

Low level deltamethrin resistance in ticks from cattle of Kerala, a south Indian state.

The deltamethrin resistance status in Rhipicephalus (Boophilus) annulatus and R. (B.) microplus ticks collected from cattle of five organized farms of Kerala, south India was evaluated. Resistance was characterized using biological (larval packet test), biochemical (esterase enzyme activity assay) and molecular tools (PCR amplification and sequencing of deltamethrin resistance-associated genes). Characterization of field isolates revealed level I resistance in ticks collected from four out of five farms. Elevated level of α/ß esterase activity was not recorded in isolates showing level I resistance. Previously reported point mutations in the carboxyl esterase (G1120A) and sodium channel (T2134A and C190A) genes were not observed in any of the field isolates. The present study showed a low level (level I) resistance is developed in the most economically important ticks infesting cattle of this state and it cautions the development of large scale resistance in future.

Acaricide activity of leaves extracts of Sambucus australis Schltdl. (Caprifoliaceae) at 2 percent on engorged females of Rhipicephalus (Boophilus) microplus / Atividade acaricida de extratos de folhas de Sambucus australis Schltdl (Caprifoliaceae) a 2 por cento sobre teleóginas de Rhipicephalus (Boophilus) microplus

Parasite infections caused by Rhipicephalus (Boophilus) microplus are responsible for the most of economic losses in producing-cattle countries in tropical and subtropical areas. Indiscriminate uses of chemical acaricides have contributed with the appearance of tick resistance to many drugs available in the marketplace, and it is a serious problem in the tick control. Flowers of Sambucus australis (South America), called "sabugueiro-from-Brazil", are used on infusions or decoctions forms in the folk medicine with diuretic, antipyretic, anti-inflammatory, and laxative purpose, also employed for treating respiratory diseases in human. The main goal of this study was to evaluate in vitro potential of S. australis leaves extracts as an acaricide agent. Ethanol extract at 70 percent has been dehydrated and a fraction was suspended in 70 percent ethanol or in distillated water at final concentration of 2 percent (0.2mg-1). Using the immersion test of engorged females the efficiency results were obtained in 34 percent and 66 percent with the leaves extract diluted in water and 70 percent ethanol, respectively. This study is the first report on acaricidal activity of S. australis against cattle tick. Further studies to determine the active metabolites in different stages of S. australis could aid to identify suitable extracts to be tested in the R. (B.) microplus control.

A infestação por Rhipicephalus (Boophilus) microplus é responsável por grandes perdas econômicas em países produtores de bovinos nas áreas tropicais e subtropicais. Usos indiscriminados de acaricidas químicos têm contribuído para o aparecimento da resistência dos ixodídeos a múltiplas drogas, representando um sério problema no controle de carrapatos. As flores da planta Sambucus australis, conhecidas pelo nome popular de sabugueiro do Brasil, são usadas, na forma de infusão ou emplastros, na medicina popular como diurético, antipirético, antiinflamatório e laxativo; também empregado no tratamento de doenças respiratórias em humanos. O objetivo principal deste trabalho foi avaliar o potencial in vitro de extratos de folhas de S. australis, como agente carrapaticida. O extrato etanólico a 70 por cento foi dessecado e retomado parte em etanol a 70 por cento ou em água destilada, numa concentração final de 2 por cento (0,2mg-1). No biocarrapaticidograma obteve-se resultados de eficácia entre 34 por cento e 66 por cento para o extrato de folhas diluído em água e etanol a 70 por cento, respectivamente. Esse estudo é o primeiro relato de ação carrapaticida de S. australis. Estudos adicionais para determinar os metabólitos ativos, em diferentes estágios de S. australis, fornecerão subsídios para identificar os extratos apropriados a serem testados no controle do R. (B.) microplus.

In vitro activity of Artemisia annua L (Asteraceae) extracts against Rhipicephalus (Boophilus) microplus / Atividade in vitro de extratos de Artemisia annua L (Asteraceae) sobre Rhipicephalus (Boophilus) microplus

The activity of plant extracts on parasites may indicate groups of substances that are potentially useful for controlling Rhipicephalus (Boophilus) microplus. The aim of the present study was to investigate the in vitro action of Artemisia annua extracts on this tick. The concentrations of the sesquiterpene lactones artemisinin and deoxyartemisinin present in plant extracts were quantified via high-performance liquid chromatography. Four extracts produced from the concentrated crude extract (CCE) were evaluated on larvae using the impregnated paper method, with readings after 24 hours of incubation. The engorged females were immersed in the CCE and in its four derived extracts for five minutes, with incubation for subsequent analysis of biological parameters. The extracts were not effective on the larvae at the concentrations tested (3.1 to 50 mg.mL-1). The CCE showed greater efficacy on engorged females (EC50 of 130.6 mg.mL-1 and EC90 of 302.9 mg.mL-1) than did the derived extracts. These results tend to confirm that the action of artemisinin on engorged females of R.(B.) microplus is conditional to their blood intake. In this case, in vitro methods would be inadequate for effective evaluation of the action of A. annua on R. (B.) microplus.

A atividade de extratos vegetais sobre parasitas pode indicar grupos de substâncias de uso potencial no controle de Rhipicephalus (Boophilus) microplus. O objetivo do presente estudo foi investigar a ação in vitro de extratos de Artemisia annua sobre esta espécie. A concentração das lactonas sesquiterpênicas artemisinina e deoxiartemisinina presentes nos extratos vegetais, foi quantificada via cromatografia líquida de alta eficiência. Quatro extratos produzidos a partir do extrato bruto concentrado (EBC) foram avaliados sobre larvas pela metodologia do papel impregnado, com leitura após 24 horas de incubação. As fêmeas ingurgitadas foram imersas por cinco minutos no EBC e nos seus quatro extratos derivados, e incubadas para posterior análise dos parâmetros biológicos. Os extratos não tiveram eficácia sobre as larvas nas concentrações avaliadas (de 3,1 a 50 mg.mL-1). O EBC apresentou melhor eficácia sobre as fêmeas ingurgitadas (CE 50 de 130,6 mg.mL-1 e CE 90 de 302,9 mg.mL-1) que os extratos derivados. Esses resultados tendem a confirmar que a ação da artemisinina sobre as fêmeas ingurgitadas de R. (B.) microplus estaria condicionada à sua ingestão através do sangue. Nesse caso, os métodos in vitro seriam inadequados para a efetiva avaliação da ação de A. annua R.(B.) microplus.

Comparison of efficiency and reproductive aptitude indexes between a reference and field strains of the cattle tick Rhipicephalus (Boophilus) microplus, in Sinaloa, Mexico / Comparação dos índices de eficiência e aptidão reprodutiva entre uma cepa de referência e outra de campo do carrapato do gado, Rhipicephalus (Boophilus) microplus, em Sinaloa, México

The objective of this work was to study the reproductive behavior of two strains of R. (B.) microplus, one wild-caught (Native) and one reference strain (Media Joya) for two years, measuring the reproductive efficiency index (REI) and reproductive aptitude index (RAI). Engorged ticks from each strain were collected monthly from February 2002 to February 2004. These were weighed and kept in the laboratory until egg-laying. Oogenic masses were individually weighed and incubated until larvae emergence. REI and RAI were calculated from each sample, grouping ticks by weight in 100 mg range classes. An analysis of variance and Duncan's multiple rank tests were carried out on the data (P < 0.05). Pearson's correlation coefficients were estimated (P < 0.01) in order to determine the relationship between engorgement weight and the weight and number of laid and hatched eggs. The Native strain showed a higher engorgement weight than Media Joya strain. In both strains REI and RAI were similar in all weight classes; with a quadratic tendency in Media Joya and linear-quadratic in the Native strain (P < 0.05). Significant differences where observed when REI and RAI of both strains were compared along the duration of the study (P < 0.01) with the Native strain performing better in both indexes, including better performance during the dry season. All parameters studied showed highly significant correlation; eggs laid and eggs hatched were notably highly correlated, 0.94 and 0.91, for Media Joya strain and Native strain respectively.

O objetivo deste trabalho foi estudar a conduta reprodutiva de duas cepas de R. (B.) microplus, uma coletada no campo (Nativa) e outra de referência (Média Jóia), por dois anos, calculando o índice de eficiência reprodutiva (REI) e o índice de aptidão reprodutiva (RAI). Foram coletados mensalmente carrapatos fêmeas engurgitadas de cada cepa no período de Fevereiro de 2002 a Fevereiro de 2004. Estas foram pesadas e conservadas no laboratório até ovipositar. As massas de ovos foram pesadas individualmente e incubadas até a eclosão das larvas. O REI e o RAI foram calculados para cada coleta, agrupando os carrapatos em classe de peso de 100 mg. Os dados foram submetidos a uma análise de variância e prova de classe múltipla de Duncan (P < 0,05). Foram calculados os coeficientes de correlação de Pearson (P < 0,01) para estimar a associação entre o peso dos carrapatos engurgitados e o peso e número de ovos postos e eclosionados. A cepa Nativa mostrou um peso ao engurgitamento mais alto do que a cepa Média Jóia. Os valores de REI e de RAI em ambas cepas foram similares em todos as classes de peso, mostrando tendência cuadrática na Média Jóia e linear-cuadrática na cepa Nativa (P < 0,05). Ambos os índices por classe de peso no engurgitamento foram sempre mais altos na cepa Nativa (P < 0,05). Os valores de REI e de RAI de ambas cepas ao longo do estudo mostraram diferenças (P < 0,01), sendo a cepa Nativa a de melhor desempenho em ambos índices, incluindo um melhor desenvolvimento durante a estação seca. Observou-se uma correlação altamente significativa em todos os parâmetros, sobressaindo o número de ovos postos com o número de ovos eclodidos, 0,94 e 0,91, para a cepa Média Jóia e a Nativa, respectivamente.