SciELO - Scientific Electronic Library Online

vol.25 issue2Hemostatic assessment of dogs associated with hepatic parasite load of Leishmania infantum chagasiGastrointestinal parasites of cats in Brazil: frequency and zoonotic risk author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand



  • text new page (beta)
  • English (pdf)
  • Article in xml format
  • How to cite this article
  • SciELO Analytics
  • Curriculum ScienTI
  • Automatic translation


Related links


Revista Brasileira de Parasitologia Veterinária

Print version ISSN 0103-846XOn-line version ISSN 1984-2961

Rev. Bras. Parasitol. Vet. vol.25 no.2 Jaboticabal Apr./June 2016  Epub June 07, 2016 

Research Note

In vitro effects of Pilocarpus microphyllus extracts and pilocarpine hydrochloride on Rhipicephalus (Boophilus) microplus

Efeitos in vitro do extrato de Pilocarpus microphyllus e do cloridrato de pilocarpina sobre Rhipicephalus (Boophilus) microplus

Karina Neoob de Carvalho Castro1  * 

David Fernandes Lima2 

Dolores Wolschick1 

Ivanilza Moreira de Andrade3 

Raimunda Cardoso dos Santos3 

Francisco José de Seixas dos Santos1 

Leiz Maria Costa Veras3 

Lívio Martins Costa-Júnior4 

1Laboratório de Doenças Parasitárias, Embrapa Meio-Norte, Parnaíba, PI, Brasil

2Anidro do Brasil Extrações S.A., Parnaíba, PI, Brasil

3Universidade Federal do Piauí – UFPI, Parnaíba, PI, Brasil

4Universidade Federal do Maranhão – UFMA, Chapadinha, MA, Brasil


The aim of this study was to assess the activity of aqueous (AE) and ethanolic extracts (EE) and pilocarpine hydrochloride, which were extracted and isolated from Pilocarpus microphyllus (Jaborandi), respectively, on Rhipicephalus (Boophilus) microplus. High performance liquid chromatography (HPLC) was performed to quantify these compounds. Larval packet and adult immersion tests were conducted with different concentrations. Five AE and EE concentrations, ranging from 6.2 to 100.0 mg mL–1, and six concentrations of pilocarpine hydrochloride, ranging from 0.7 to 24.0 mg mL–1, were tested. The lethal concentration (LC50) of each extract for larvae and engorged females was calculated through Probit analysis. The concentration of pilocarpine hydrochloride obtained from the EE and the AE was 1.3 and 0.3% (m/m), respectively. Pilocarpine hydrochloride presented the highest acaricidal activity on larvae (LC50 2.6 mg mL–1) and engorged females (LC50 11.8 mg mL–1) of R.(B.) microplus, followed by the EE which presented LC50 of 56.4 and 15.9 mg mL–1, for larvae and engorged females, respectively. Such results indicate that pilocarpine hydrochloride has acaricidal activity, and may be the primary compound responsible for this activity by P. microphyllus EE.

Keywords:  Acaricide; tick; control; pilocarpine hydrochloride; Pilocarpus micropyllus


O objetivo desse estudo foi avaliar a atividade dos extratos aquoso (AE) e etanólico (EE) e do cloridrato de pilocarpina, que foram, respectivamente, extraídos e isolado de Pilocarpus microphyllus (Jaborandi), sobre Rhipicephalus (Boophilus) microplus. Cromatografia líquida de alta eficiência foi realizada para quantificação dos compostos. Testes de pacote de larvas e de imersão de adultos foram realizados com diferentes concentrações. Cinco concentrações do AE e EE variando de 6,2 a 100,0 mg mL–1 e seis concentrações do cloridrato de pilocarpina variando de 0,7 a 24,0 mg mL–1 foram testadas. A concentração letal (CL50) de cada extrato para larvas e fêmeas ingurgitadas foi estimada por meio da análise Probit. A concentração de cloridrato de pilocarpina obtida do EE e AE foi de 1,3 e 0,3% (m/m), respectivamente. O cloridrato de pilocarpina apresentou a maior atividade carrapaticida sobre larvas (CL50 2,6 mg mL–1) e fêmeas ingurgitadas (CL50 11,8 mg mL–1) de R. (B.) microplus, seguido do EE que apresentou CL50 de 56,4 e 15,9 mg mL–1, para larvas e fêmeas ingurgitadas, respectivamente. Tais resultados indicam que o cloridrato de pilocarpina apresenta atividade carrapaticida e pode ser o principal responsável pela atividade acaricida do EE de P. microphyllus.

Palavras-chave:  Acaricida; carrapato; controle; cloridrato de pilocarpina; Pilocarpus micropyllus


Rhipicephalus (Boophilus) microplus is the main cattle tick throughout the tropics (GRISI et al., 2014). Infestation by this parasite results in losses, mainly due to the decreased milk and meat production, and also due to the transmission of pathogens, which cause babesiosis and anaplasmosis (PETER et al., 2005; GRISI et al., 2014).

The control of R. (B.) microplus is conducted through chemical acaricides, and this process is often performed indiscriminately (FURLONG et al., 2003). The indiscriminate use of these products has contributed to the contamination of the environment and products of animal origin such as milk and meat. It has also led to the selection of ticks that are resistant to all active compounds that are normally found in the market (CASTRO-JANER et al., 2010; MILLER et al., 2013). In many countries, Brazil included, the acaricide-resistant tick population has been increasing, to the point that few acaricides have efficacy of more than 75% of the tick population (GRAF et al., 2004; MENDES et al., 2011). On the other hand, the results of certain studies suggest that environmentally friendly acaricides can be obtained from plants (GHOSH et al., 2011; GARCIA et al., 2012). As an alternative, plant extracts are used to manage animal health and parasite control in several regions in the world (GITHIORI et al., 2006; TAMIRU et al., 2013; NYAHANGARE et al., 2015).

Currently, several experimental studies have been assessing the anti-parasitic activity of medicinal plants (BORGES et al., 2011; DOMINGUES et al., 2013; LIMA et al., 2014). However, few plant extracts or compounds are produced on an industrial scale, which would enable their use for the control of ticks in the field.

The therapeutic use of Jaborandi (Pilocarpus sp.) has long been known by indigenous tribes from northern Brazil as an antidote for venoms and toxins. Currently, this medicinal plant is intensively used in the pharmaceutical industry (VITAL & ACCO, 2006). The genus Pilocarpus belongs to the family Rutaceae and is distributed in Brazil from the Amazon region as far as Rio Grande do Sul state (JOSEPH, 1967). This genus has many species of economic and medical importance, among which is Pilocarpus microphyllus (CORRÊA, 1969). Over the last three decades this specie has been considered one of the most important in the Brazilian flora for the production of medications. There are approximately 15 million of Jaborandi plants in cultivation and these produce about 10,000 kg of leaves per year (PINHEIRO, 2002, 2006). It is important to note that P. microphyllus has alkaloids that are viewed as promising sources in the quest for anti-parasitic agents (LEITE et al., 2011; VÉRAS et al., 2012).

Therefore, this study aimed at investigating the effects of aqueous and ethanolic extracts and pilocarpine hydrochloride obtained from P. microphyllus on the larvae and engorged females of the R.(B.) microplus tick.

Materials and Methods

Plant material

Leaves of P. microphyllus were collected at Anidro do Brasil Extrações S.A. pharmaceutical chemical company, located in the city of Parnaíba, Piauí, Brazil. Reference plant specimens were deposited in the Herbarium (CEN) of the Embrapa Recursos Genéticos e Biotecnologia, under registration number 81.098.

The leaves were sun dried until they reached ≤ 15% moisture content, and ground in a knife mill, so the aqueous (AE) and ethanolic (EE) extracts of P. microphyllus could be prepared. The AE was obtained through a 20-minute infusion with distilled water at 95 °C. The EE was obtained through maceration in ethanol (analytical grade) for seven days, with solvent changes. This material was filtered and concentrated in a rotary evaporator, and then diluted in 50% ethanol and 3% DMSO. Pilocarpine hydrochloride (100% purity) from P. microphyllus was kindly donated by Anidro do Brasil Extrações S.A. company (Parnaíba, PI, Brazil).

Chemical analysis

The pilocarpine assay % (m/m) in the AE and EE was determined through high performance liquid chromatography (HPLC) with a Merck-Hitachi Lachron Elite L-2000 chromatograph; mobile phase: 5% KH2PO4 and pH 2.5; Merck/ Lichrospher® 60 RP select B chromatography column (250 × 4.6 mm, 5 µm); 50 °C oven; UV 216 nm detector; 1 mL/min. flow; external standard of pilocarpine hydrochloride as the reference (VÉRAS et al., 2013).

Tick preparation

Engorged R. (B.) microplus females were collected from naturally infested calves. They were washed with water and dried with paper towels. A number of the engorged females were kept in a BOD chamber (27 °C and RH > 80%) until oviposition of the larvae (for the larval packet test) was finished. Previously, acaricide tests were performed which demonstrated resistance of the tick population to amidinic and pyrethroid compounds (personal information).

Preparation of the dilutions

The AE and the EE were prepared at concentrations of 6.2, 12.5, 25.0, 50.0 and 100.0 mg mL–1 and pilocarpine hydrochloride at concentrations of 0.7, 1.5, 3.0, 6.0, 12.0 and 24.0 mg mL–1. The negative controls consisted of the solvents used in AE and EE dilutions, which were ultrapure water (C1) and 50% ethanol + 3% DMSO (C2), respectively. As the positive control, a mixture of 0.18 mg mL–1 cypermethrin, 0.30 mg mL–1 chlorpyrifos, and 0.012 mg mL–1 citronellal (Colosso®, Ouro Fino, São Paulo) was used. This solution was diluted at a 0.125% concentration in ultrapure water. All tests were replicated three times.

Larval packet test

The larval packet test was used following the method of Stone & Haydock (1962) and the Food and Agriculture Organization of the United Nations (FAO, 1971). Approximately 100 14-to-21-day-old larvae were placed between two filter papers (2 × 2 cm) forming a sandwich, after which they were impregnated with the treatments. Each “sandwich” was introduced in a filter paper envelope, and then sealed, identified, and incubated at 27 °C with RH > 80% (LEITE, 1988). Living and dead larvae were counted 24 hours later, and mortality was calculated from the arithmetic average of three replicates.

Adult immersion test

The adult immersion test of engorged R. (B.) microplus females was conducted according to the method described by Drummond et al. (1973). After they were selected based on their mobility, body integrity, and size (≥ 4.5 mm), engorged females were weighed and distributed in ten-specimen groups with similar weights. The weight of engorged females ranging between 170 to 210 mg (BENNETT, 1974). The groups of females were immersed in solutions containing the several treatments for 5 minutes, and then dried in paper towels and stored in a chamber at 27 °C and RH > 80% for 18 days. After that period, the egg mass was weighed, transferred to adapted syringes, and incubated for 20 days (27 °C and RH > 80%). Hatchability was estimated from the average number of eggs and larvae. The egg production index (EPI), oviposition reduction (OR), reproductive efficiency (RE), and product effectiveness (PE) were calculated according to the following formulas: EPI = (weight of eggs/engorged females) × 100 (BENNETT, 1974); OR = (control EPI − treated EPI/control EPI) × 100 (ROULSTON et al., 1968); RE= (Egg mass weight × % of eclosion/weight of the mass of females) × 20,000; and PE = (control RE – treated RE)/(control RE × 100) (DRUMMOND et al., 1973).

This study was performed with the approval of the Ethics Committee for Animal Experimentation of the Federal University of Maranhão under approval number 23115018061.

Statistical analysis

The lethal concentration to kill 50% of the studied population (LC50) of each extract for larvae and engorged females was calculated through Probit analysis with GraphPad Prism 6.0 software. The extract was considered to be significantly different from another one when the 95% confidence interval of LC50 was not overlapped (RODITAKIS et al., 2005). The differences among the concentrations of mortality against larvae and engorged females and the index RO, hatchability and EP were analyzed by F test of ANOVA followed by Tukey test (p<0.05).

Results and Discussion

The concentrations of pilocarpine obtained from the EE and the AE were 1.3 and 0.3% (m/m), respectively. The presence of active substances in the plant, even in small proportions or associated to other substances also in small proportions, may be responsible for the biological activity of some extracts (LIMA et al., 2014).

In this study, the EE at 100 mg mL–1 resulted in a 30.3% mortality for R. (B.) microplus larvae (Table 1). To increase mortality, higher concentrations would be necessary making its use difficult in the field (LC50 56.4 mg mL–1; 95% CI 38.98 – 81.66) (Table 2). In regards to the R. (B.) microplus engorged females, the efficiency of EE at 100 mg mL–1 was 79.2% (LC5015.9 mg mL–1; 95% CI 14.30-17.88), which demonstrates a significant effect on their reproductive capabilities (Table 1). This effect is mainly due to reduced oviposition (76.3%). There was no significant influence on the hatchability of eggs (85.5%) (Table 1). When the AE was used against larvae and engorged R. (B.) microplus females, no parameter was significantly different, and no acaricidal activity was found (Table 1).

Table 1 Action of the aqueous (AE) and etanólico (EE) extract of Pilocarpus microphyllus and of pilocarpine hydrochloride (PC) on larvae (mortality) and engorged female (reduction of oviposition, hatchability and efficacy of the product) of Rhipicephalus (Boophilus) microplus

Treatment Concentration (mg mL–1) Larvae Engorged Females
Mortality (%) Weight of females (g) Weight of eggs (g) Reduction of oviposition (%) Hatchability (%) Efficacy of the product (%)
C1 - 0.7±1.3a 1.948±0.027 1.068±0.092 - 95.3±1.1a -
C2 - 0.0±0.0a 1.887±0.018 1.131±0.025 - 96.4±1.2ª -
AE 6.2 0.0±0.0a 1.923±0.009 1.153±0.007 0.9±0.5ª 94.0±3.6ª 3.4±3.3ª
12.5 0.0±0.0a 1.922±0.020 1.135±0.025 2.4±1.1ª 94.9±0.5ª 3.9±0.6ª
25.0 0.0±0.0a 1.916±0.017 1.118±0.037 3.5±2.8ª 96.2±0.3ª 3.8±2.5ª
50.0 0.0±0.0a 1.925±0.017 1.140±0.024 2.1±2.6ª 96.2±1.7ª 2.3±1.6ª
100.0 1.0±1.7ª 1.925±0.008 1.157±0.004 0.7±0.2ª 94.3±1.2ª 2.9±1.4ª
EE 6.2 1.5±1.7ª 1.891±0.002 1.000±0.023 11.8±2.1ª,b 92.3±2.5ª 14.5±1.8ª,b
12.5 2.6±2.1ª 1.877±0.014 0.842±0.092 25.1±8.5b 94.2±1.6ª 26.1±7.1b,c
25.0 3.5±2.3ª 1.870±0.008 0.378±0.113 66.3±10.0c 91.3±2.9ª 67.6±10.4d
50.0 9.4±1.4ª 1.868±0.018 0.377±0.038 66.4±3.4c 87.8±4.2ª,b 68.9±4.0d
100.0 30.3±8.3b 1.887±0.010 0.268±0.157 76.3±13.9d 85.5±5.3ª,b 79.2±11.4d,e
PC 0.7 10.2±6.1ª 1.956±0.009 1.015±0.078 7.2±6.9ª 93.7±3.4ª 10.4±3.3ª,b
1.5 32.7±14.8b 1.962±0.029 1.028±0.049 6.2±5.8ª 90.7±0.9ª 12.4±4.7ª,b
3.0 58.8±9.0c 1.983±0.007 1.017±0.033 8.2±3.2ª 95.5±1.4ª 9.6±2.1ª,b
6.0 88.4±7.6d 1.930±0.025 0.947±0.111 12.3±9.4ª,b 75.6±6.0b 31.4±11.6b,c
12.0 99.0±0.9d 1.980±0.009 0.666±0.116 30.4±12.3b 74.8±13.8b 43.2±19.1c
24.0 96.6±2.1d 1.978±0.014 0.219±0.037 77.1±3.7c,d 26.8±6.8c 93.2±2.8e

Values represent average ± standard deviation; C1: Negative control (mili-q water); C2: Negative control (ethanol 50% + DMSO 3%); Means with the same letter, for each treatment, are not statistically different (p ≤ 0.05).

Table 2 Lethal concentration of Pilocarpus microphyllus leaves ethanolic extract and of pilocarpine hydrochloride against Rhipicephalus (Boophilus) microplus larvae and engorged females. 

Treatment LC50 (mg mL–1) IC 95% R2
Ethanolic extract 56.4c 38.98-81.66 0.90
pilocarpine hydrochloride 2.6ª 2.36-2.97 0.97
Engorged female
Ethanolic extract 15.9b 14.30-17.88 0.92
pilocarpine hydrochloride 11.8b 9.60-14.41 0.88

LC50 concentration (mg/mL) at which 50% of the R. (B.) microplus died; IC 95% - Confidence limits at 95% probability; R2 = Coeficient of determination.

The assessment of pilocarpine hydrochloride on larvae obtained a maximum mortality (99.0%) (Table 1) at 12 mg mL–1 (LC50 2.6 mg mL–1; 95% IC 2.36-2.97) (Table 2). At 24 mg mL–1, pilocarpine hydrochloride was found to be highly effective (93.2%) in its effect on engorged females, (LC50 11.8 mg mL–1; 95% IC 9.60-14.41). This was due to low oviposition (77.1%) and to low hatchability (26.8%) (Table 1), which demonstrates the reduced egg fertility resulting from reproductive impairment. The number of offspring was consequently reduced.

In this study, the effect of isolated pilocarpine hydrochloride on R. (B.) microplus larvae and engorged females was found to be higher than the effect of the EE of P. microphyllus, which contained 1.3% of the compound. In contrast, Chungsamarnyart & Jansawan (1996), when assessing other species in the Rutaceae family, observed a significantly reduced effect of the d-limonene constituent on R. (B.) microplus larvae and females, compared to the effect of oils from Citrus spp. species, from which it originated, indicating, in that case, a possible synergy among its compounds.

The effect of pilocarpine hydrochloride was shown to be similar to that of the positive control (a mixture of cypermethrin, chlorpyrifos, and citronellal) on larvae (mortality of 100%) in the last three concentrations (6, 12 and 24 mg mL–1) and engorged females (efficacy of 98,2%) in the last concentration (24 mg mL–1) for the three parameters (reduction of oviposition, hatchability and efficacy of the product) (Table 1). These results indicate the potential of that substance in the control of R. (B.) microplus. An advantage in the use of plant extracts and their compounds is in the possibility of controlling cattle ticks in organic production systems, where the use of synthetic pesticides is prohibited. However, this is only true when these extracts do not cause adverse reactions to the host and workers, and do not leave residues in products of animal origin. It is also an alternative way to control resistant populations (BORGES et al., 2011).

Synergistic combinations are also used to reduce the dosage of substances and the risk of parasites developing resistance (TRIPATHI et al., 2009). The effect is enhanced by the interaction of its various compounds (CHAGAS et al., 2002). The resistance develops more slowly when extracts or compounds are used, due to the mixture of active substances with different mechanisms (MULLA & SU, 1999). Therefore, an extract-based acaricide, when compared to one composed of a single main substance, has an edge in terms of both reduced resistance and cost.

P. microphyllus, from which pilocarpine is extracted, is native and widely spread in Piauí, Maranhão and Pará states of Brazil (PINHEIRO, 1997). Currently, the annual industrial production of Jaborandi leaves is 10,000 kg, of which approximately 1% is pilocarpine (VÉRAS et al., 2013) demonstrating the production potential of this medicinal species and its pilocarpine alkaloid. It is extremely important to emphasize that Brazil is the only supplier of this active pharmaceutical ingredient (API) for the international pharmaceutical industry (BRANDÃO et al., 2008) and studies with P. microphyllus alkaloids and its industrial applications are of fundamental importance for a large number of economically precarious communities whose main source of income is the species harvest in underdeveloped regions of northeastern Brazil (LIMA et. al., 2015).

Pilocarpine is a cholinergic alkaloid that, when connected to muscarinic receptors, acts on the parasympathetic nervous system and results in smooth muscle contraction. Therefore, it stimulates secretory glands, relaxes sphincters, stimulates peristalsis, and influences blood pressure in humans (PAPPANO, 2006). There is evidence that muscarinic receptors are present in the synganglion of R. (B.) microplus (TURBERG et al., 1996). Peripheral nerves emerge from the synganglion, during the several life stages of ticks, and they are responsible for the innervation of muscles, sensory structures, tegmen, and several internal organs (SONENSHINE, 1991). Pilocarpine may have inhibited engorged female reproduction and killed the larvae, as observed in this study, due to the excessive stimulation of smooth muscles in digestive, reproduction, or circulatory organs, thus producing a physiological imbalance in R. (B.) microplus.

Pilocarpine is also the most widely used agonist to induce salivation in ticks, and the salive obtained is used to develop a vaccine against them (RIBEIRO et al., 2004). However, no previous study has reported pilocarpine leading to the inhibition of the reproduction of females or the mortality of R. (B.) microplus larvae.

Our results indicate that the EE of P. microphyllus has significant effects on R. (B.) microplus larvae and engorged females and that pilocarpine hydrochloride may be one of the main agents responsible for these effects. Based on our results, we infer that higher pilocarpine contents in the EE may enhance the effects obtained on R. (B.) microplus. The use of certain processes to optimize extraction of the P. microphyllus EE may result in the possibly more efficient recovery of extracts with higher concentrations of pilocarpine alkaloid. The acidification of P. microphyllus leaves and the further extraction with polar solvents such as water and ethanol favor the extraction of total alkaloids that can be subsequently submitted to fractionation and prepurification processes (VÉRAS et al., 2013). Due to the high cost of obtaining isolated pilocarpine hydrochloride, obtaining extracts with a higher content of that alkaloid is an alternative that could possibly be used as a low-cost acaricide. Complementary studies should be performed to establish the safety of the use of these extracts in animals.


The ethanolic extracts of P. microphyllus and pilocarpine hydrochloride have significant effects on R. (B.) microplus larvae and, most notably, on engorged females. These results suggest the possibility of using these extracts as acaricides, conducting new studies with optimized extracts and fractions of P. microphyllus with a higher pilocarpine content and with the goal of enhancing its effects on R. (B.) microplus. Furthermore, studies that add potential biotechnological value to this species will benefit underdeveloped communities whose main source of income is the harvest of leaves from native species in the states of Piauí, Maranhão and Pará in northeast Brazil.


The authors would like to thank Mr. João Batista Alves de Souza and Mr. Antônio de Almeida Paz Neto for their valuable cooperation with the experiments.


Bennett GF. Oviposition of (Canestrini) (Acarida: Ixodidae). I. Influence of tick size on egg production. Boophilus microplusAcarologia 1974; 16(1): 52-61. PMid:4463680. [ Links ]

Borges LMF, Sousa LAD, Barbosa CS. Perspectivas para o uso de extratos de plantas para o controle do carrapato de bovinos ( RhipicephalusBoophilus) microplus.Rev Bras Parasitol Vet 2011; 20(2): 89-96. PMid:21722481. [ Links ]

Brandão MG, Zanetti NN, Oliveira P, Grael CF, Santos AC, Monte-Mór RL. Brazilian medicinal plants described by 19th century European naturalists and in the Official Pharmacopoeia. J Ethnopharmacol 2008; 120(2): 141-148. PMid:18762237. [ Links ]

Castro-Janer E, Martins JRS, Mendes MCD, Namindome AB, Klafke GMB, Schumaker TTS. Diagnoses of fipronil resistance in Brazilian cattle ticks (Rhipicephalus (Boophilus) microplus) using in vitro larval biossays. Vet Parasitol 2010; 173(3-4): 300-306. PMid:20688434. [ Links ]

Chagas ACS, Passos WM, Prates HT, Leite RC, Furlong J, Fortes ICP. Efeito acaricida de óleos essenciais e concentrados emulsionáveis de Eucalyptus spp em Boophilus microplus.Braz J Vet Res Anim Sci 2002; 39(5): 247-253. [ Links ]

Chungsamarnyart N, Jansawan W. Acaricidal activity of peel oil of spp. on CitrusBoophilus microplus.Kasetsart J 1996; 30: 112-117. [ Links ]

Corrêa MP. Dicionário das plantas úteis do Brasil e exóticas cultivadas. Rio de Janeiro: Ministério da Agricultura; 1969. 765 p. vol. 4. [ Links ]

Domingues LF, Giglioti R, Feitosa KA, Fantatto RR, Rabelo MD, Oliveira MCS, et al. activity of pineapple extracts (. In vitroAnanas comosus, Bromeliaceae) on Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)Exp Parasitol 2013; 134(3): 400-404. PMid:23541882. [ Links ]

Drummond RO, Ernst SE, Trevino JL, Gladney WJ, Graham OH. and . Boophilus annulatusB. microplus: laboratory tests for insecticidesJ Econ Entomol 1973; 66(1): 130-133. PMid:4690254. [ Links ]

Food and Agriculture Organization of the United Nations – FAO. Recommended methods for the detection and measurement of resistance of agricultural pests to pesticides. Tentative methods for larvae of cattle tick Boophilus spp. Rome: FAO; 1971. p. 15-18. FAO method. vol. 7. FAO Plant Protection Bulletin. [ Links ]

Furlong J, Martins JR, Prata MC. Carrapato dos bovinos: controle estratégico nas diferentes regiões brasileiras. Juiz de Fora: Embrapa Gado de Leite; 2003. 5 p. vol. 36. Comunicado Técnico. [ Links ]

Garcia MV, Matias J, Barros JC, Lima DP, Lopes RS, Andreotti R. Chemical identification of Tagetes minuta Linnaeus (Asteraceae) essential oil and its acaricidal effect on ticks. Rev Bras Parasitol Vet 2012; 21(4): 405-411. PMid:23295821. [ Links ]

Ghosh S, Sharma AK, Kumar S, Tiwari SS, Rastogi S, Srivastava S, et al. In vitro and in vivo efficacy of Acorus calamus extract against Rhipicephalus (Boophilus) microplus.Parasitol Res 2011; 108(2): 361-370. PMid:20886235. [ Links ]

Githiori JB, Athanasiadou S, Thamsborg SM. Use of plants in novel approaches for control of gastrointestinal helminths in livestock with emphasis on small ruminants. Vet Parasitol 2006; 139(4): 308-320. PMid:16725262. [ Links ]

Graf JF, Gogolewski R, Leach-Bing N, Sabatini GA, Molento MB, Bordin EL, et al. Tick control: an industry point of view. Parasitology 2004;129(7 Suppl): 427-442. PMid:15938522. [ Links ]

Grisi L, Leite RC, Martins JRS, Barros ATM, Andreotti R, Cançado PHD, et al. Reassessment of the potential economic impact of cattle parasites in Brazil. Rev Bras Parasitol Vet 2014; 23(2): 150-156. PMid:25054492. [ Links ]

Joseph CJ. Revisão sistemática do gênero (ssp. brasileiras). PilocarpusMecânica Popular 1967; 40: 1-9. [ Links ]

Leite JRSA, Miura LMCV, Lima DF, Carneiro SMP, Carvalho FAA, Moraes J, et alProcesso de obtenção da epiisopiloturina e sua aplicação no combate à infecções parasitáriasPatent PI 0904110-9 A22011May31 [ Links ]

Leite RC. [Boophilus microplus (Canestrini, 1887): susceptibilidade, uso atual e retrospectivo de carrapaticidas em propriedades das regiões fisiogeográficas da baixada do Grande-Rio e Rio de Janeiro: uma abordagem epidemiológicaThesis]. Itaguaí: Universidade Federal Rural do Rio de Janeiro; 1988. [ Links ]

Lima AS, Sousa-Filho JGN, Pereira SG, Guillon G, Santos LS, Costa-Júnior LM. Acaricide activity of different extracts from fruits against Piper tuberculatumRhipicephalus microplus.Parasitol Res 2014; 113(1): 107-112. PMid:24221883. [ Links ]

Lima DF, Silva RAO, Marques LGA, Véras LMC, Simões ERB, Leite JRSA, et al. Technological Forecasting of Jaborandi (Pilocarpus microphyllus): economically important specie in the north and northeast of Brazil. GEINTEC 2015; 5(1): 1626-1638. [ Links ]

Mendes MC, Lima CK, Nogueira AH, Yoshihara E, Chiebao DP, Gabriel FH, et al. Resistance to cypermethrin, deltamethrin and chlorpyriphos in populations of (Acari: Ixodidae) from small farms of the State of São Paulo, Brazil. Rhipicephalus (Boophilus) microplusVet Parasitol 2011; 178(3-4): 383-388. PMid:21306827. [ Links ]

Miller RJ, Almazán C, Ortíz-Estrada M, Davey RB, George JE, De León AP. First report of fipronil resistance in Rhipicephalus (Boophilus) microplus of Mexico. Vet Parasitol 2013; 191(1-2): 97-101. PMid:23026557. [ Links ]

Mulla MS, Su T. Activity and biological effects of neem products against arthropods of medical and veterinary importance. J Am Mosq Control Assoc 1999; 15(2): 133-152. PMid:10412110. [ Links ]

Nyahangare ET, Mvumi BM, Mutibvu T. Ethnoveterinary plants and practices used for ecto-parasite control in semi-arid smallholder farming areas of Zimbabwe. J Ethnobiol Ethnomed 2015; 11(1): 30. PMid:25925402. [ Links ]

Pappano AJ. Drogas ativadoras dos receptores colinérgicos e inibidores da colinesterase. In: Katzung BG. Farmacologia básica e clínica. 9th ed. Rio de Janeiro: Guanabara Koogan; 2006. p. 79-90. [ Links ]

Peter RJ, Van den Bossche P, Penzhorn BL, Sharp B. Tick, fly, and mosquito control-Lessons from the past, solutions for the future. Vet Parasitol 2005; 132(3-4): 205-215. PMid:16099104. [ Links ]

Pinheiro CUB. Jaborandi ( sp., Rutaceae): a wild species and its rapid transformation into a crop. PilocarpusEcon Bot 1997; 51(1): 49-58. [ Links ]

Pinheiro CUB. Extrativismo, cultivo e privatização do jaborandi ( stapf ex Holm.; Rutaceae) no Maranhão, Brasil. Pilocarpus microphyllusActa Bot Bras 2002; 16(2): 141-150. [ Links ]

Pinheiro CUB. Extractivism, domestication, and privatization of a native plant resource: the case of Jaborandi (. In: Pilocarpus microphyllus Stapf ex Holmes) in Maranhão, BrazilPosey DA, Balick MJ. Human impacts on Amazonia: The role traditional ecological knowledge in conservation and development. New York: Columbia University Press; 2006. p. 210-221. [ Links ]

Ribeiro JMC, Zeidner NS, Ledin K, Dolan MC, Mather TN. How much pilocarpine contaminates pilocarpine-induced tick saliva? Med Vet Entomol 2004; 18(1): 20-24. PMid:15009442. [ Links ]

Roditakis E, Roditakis NE, Tsagkarakou A. Insecticide resistance in Bemisia tabaci (Homoptera: Aleyrodidae) populations from Crete. Pest Manag Sci 2005; 61(6): 577-582. PMid:15712366. [ Links ]

Roulston WJ, Schnitzerling HJ, Schuntner CA. Acetylcholinesterase insensitivity in the Biarra strain of the cattle tick Boophilus microplus, as a cause of resistance to organophosphorus and carbamate acaricides. Aust J Biol Sci 1968; 21(4): 759-767. PMid:5680996. [ Links ]

Sonenshine DE. Biology of ticks. New York: Oxford University Press; 1991. [ Links ]

Stone BF, Haydock KP. A method for measuring the acaricide susceptibility of the cattle tick (Can.). Boophilus microplusBull Entomol Res 1962; 53(3): 563-578. [ Links ]

Tamiru F, Terfa W, Kebede E, Dabessa G, Roy RK, Sorsa M. Ethnoknowledge of plants used in veterinary practices in Dabo Hana District, West Ethiopia. J Med Plant Res 2013; 7(40): 2960-2971. [ Links ]

Tripathi AK, Upadhyay S, Bhuiyan M, Bhattacharya PRA. A review on prospects of essential oils as biopesticide in insect-pest management. J Pharmacognosy Phytother 2009; 1(5): 52-63. [ Links ]

Turberg A, Schroder I, Wegener S, Londershausen M. Presence of muscarinic acetylcholine receptors in the cattle tick and in epithelial tissue culture cells of Boophilus microplusChironomus tentans.Pestic Sci 1996; 48(4): 389-398.<389::AID-PS502>3.0.CO;2-Q. [ Links ]

Véras LMC, Cunha VRR, Lima FCDA, Guimarães MA, Vieira MM, Campelo YDM, et al. Industrial scale isolation, structural and spectroscopic characterization of epiisopiloturine from Pilocarpus microphyllus stapf leaves: a promising Alkaloid against Schistosomiasis. PLoS One 2013; 8(6): e66702. PMid:23840522. [ Links ]

Véras LMC, Guimarães MA, Campelo YDM, Vieira MM, Nascimento C, Lima DF, et al. Activity of epiisopiloturine against Schistosoma mansoni.Curr Med Chem 2012; 19(13): 2051-2058. PMid:22420337. [ Links ]

Vital MABF, Acco A. Agonistas e antagonistas colinérgicos. In: Spinosa HS, Górniak SL, Bernardi MM. Farmacologia aplicada à medicina veterinária. Rio de Janeiro: Guanabara Koogan; 2006. cap. 7, p. 84-85. [ Links ]

Received: September 25, 2015; Accepted: September 29, 2015

*Corresponding author: Karina Neoob de Carvalho Castro. Embrapa Meio-Norte, Rodovia BR 343, Km 35, CEP 64200-970, Parnaíba, PI, Brasil. e-mail:

Creative Commons License This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.