Chemical composition, in vitro larvicidal and antileishmanial activities of the essential oil from Citrus reticulata Blanco fruit peel

Composição química, atividades larvicida e leishmanicida in vitro do óleo essencial da casca do fruto de Citrus reticulata Blanco

A. C. S. D. Oliveira C. C. Fernandes L. S. Santos A. C. B. B. Candido L. G. Magalhães M. L. D. Miranda About the authors

Abstract

Numerous studies have investigated the chemical composition and biological activities of essential oils from different Citrus species fruit peel, leaves and flowers. This paper aims to investigate the chemical composition, larvicidal and antileishmanial activities of essential oil from Citrus reticulata fruit peel (CR-EO). CR-EO was obtained by hydrodistillation in a Clevenger-type apparatus and its chemical composition was analyzed by GC-MS and GC-FID. Limonene (85.7%), ɣ-terpinene (6.7%) and myrcene (2.1%) were identified as its major components. CR-EO showed high activity against promastigote forms of Leishmania amazonensis (IC50 = 8.23 µg/mL). CR-EO also exhibited high larvicidal activity against third instar Aedes aegypti larvae at a lethal concentration (LC50 = 58.35 µg/mL) and 100% mortality at 150 µg/mL. This study suggests, for the first time, the potential use of CR-EO against this important mosquito-borne viral disease caused by the genus Aedes.

Keywords:
Leishmania amazonensis; Aedes aegypti; limonene

Resumo

Numerosos estudos têm investigado a composição química e as atividades biológicas de óleos essenciais extraídos de cascas dos frutos, folhas e flores de diferentes espécies de Citrus. Este trabalho tem como objetivo investigar a composição química e as atividades larvicida e leishmanicida in vitro do óleo essencial das cascas dos frutos de Citrus reticulata (CR-EO). CR-EO foi obtido pela técnica de extração em aparelho Clevenger e sua composição química foi determinada por CG-EM e CG-DIC. Limoneno (85,7%), ɣ-terpineno (6,7%) and mirceno (2,1%) foram identificados como os constituintes majoritários. CR-EO mostrou alta atividade contra as formas promastigota de Leishmania amazonensis (CI50 = 8,23 µg/mL). CR-EO também exibiu alta atividade larvicida contra as larvas do terceiro estágio do Aedes aegypti com concentração letal (CL50 = 58,35 µg/mL) e mortalidade de 100% em 150 µg/mL. Este estudo sugere, pela primeira vez, o uso potencial de CR-EO contra esta importante doença viral transmitida por mosquitos do gênero Aedes.

Palavras-chave:
Leishmania amazonensis; Aedes aegypti; limoneno

1. Introduction

Concern for the control and fight against Aedes aegypti, the mosquito that is the main vector of a severe hemorrhagic disease known as dengue, has been latent (Wankhar et al., 2015WANKHAR, W., SRINIVASAN, S. and RATHINASAMY, S., 2015. HPTLC analysis of Scoparia dulcis Linn (Scrophulariaceae) and its larvicidal potential against dengue vector Aedes aegypti. Natural Product Research, vol. 29, no. 18, pp. 1757-1760. http://dx.doi.org/10.1080/14786419.2014.999060. PMid:25573588.
http://dx.doi.org/10.1080/14786419.2014....
). The number of dengue cases has reached about 400 million per year. In the last 50 years, this disease has been endemic in 128 countries, while 36 countries are considered dengue-free ones. The highest prevalences are found in countries in the Americas, Asia and Africa, i. e., 3.97 billion inhabitants are exposed to the risk of infection (Brady et al., 2012BRADY, O.J., GETHING, P.W., BHATT, S., MESSINA, J.P., BROWNSTEIN, J.S., HOEN, A.G., MOYES, C.L., FARLOW, A.W., SCOTT, T.W. and HAY, S.I., 2012. Refining the global spatial limits of dengue virus transmission by evidence-based consensus. PLoS Neglected Tropical Diseases, vol. 6, no. 8, e1760. http://dx.doi.org/10.1371/journal.pntd.0001760. PMid:22880140.
http://dx.doi.org/10.1371/journal.pntd.0...
). In addition to the dengue virus, the vector may transmit other diseases known all over the world as chikungunya and zika (Neves et al., 2017NEVES, I.A., REZENDE, S.R.F., KIRK, J.M., PONTES, E.G. and CARVALHO, M.G., 2017. Composition and larvicidal activity of essential oil of Eugenia candolleana DC. (Myrtaceae) against Aedes aegypti. Revista Virtual de Química, vol. 9, no. 6, pp. 2305-2315. http://dx.doi.org/10.21577/1984-6835.20170138.
http://dx.doi.org/10.21577/1984-6835.201...
).

Leishmaniasis is another worrisome disease worldwide which is also caused by the infected vector known as “straw mosquito” (Silva et al., 2020SILVA, F.F.A., FERNANDES, C.C., OLIVEIRA, G.A., CANDIDO, A.C.B.B., MAGALHÃES, L.G., VIEIRA, T.M., CROTTI, A.E.M., SILVA, C.A. and MIRANDA, M.L.D., 2020. In vitro antileishmanial and antioxidant activities of essential oils from different parts of Murraya paniculata (L.) Jack: a species of Rutaceae that occur in the Cerrado bioma in Brazil. Australian Journal of Crop Science, vol. 14, no. 2, pp. 347-353. http://dx.doi.org/10.21475/ajcs.20.14.02.p1966.
http://dx.doi.org/10.21475/ajcs.20.14.02...
). The estimate is that 2 million new cases of leishmaniasis occur annually and that between 15 and 20 million people have got the disease in the world (Moreira et al., 2019MOREIRA, R.R.D., SANTOS, A.G., CARVALHO, F.A., PEREGO, C.H., CREVELIN, E.J., CROTTI, A.E.M., COGO, J., CARDOSO, M.L.C. and NAKAMURA, C.V., 2019. Antileishmanial activity of Melampodium divaricatum and Casearia sylvestris essential oils on Leishmania amazonensis. Revista do Instituto de Medicina Tropical de São Paulo, vol. 61, pp. e33. http://dx.doi.org/10.1590/s1678-9946201961033. PMid:31269109.
http://dx.doi.org/10.1590/s1678-99462019...
). In the search for plants with therapeutic potential, species of the genus Citrus not only have economic importance, but also produce bioactive essential oils (EOs) which have high value in perfume, food and beverage industries (Dosoky and Setzer, 2018DOSOKY, N.S. and SETZER, W.N., 2018. Biological activities and safety of Citrus spp. Essential oils. International Journal of Molecular Sciences, vol. 19, no. 7, pp. 1966. http://dx.doi.org/10.3390/ijms19071966. PMid:29976894.
http://dx.doi.org/10.3390/ijms19071966...
).

EOs and extracts from Citrus reticulata fruit peel and leaves have several biological applications, such as antimicrobial, antioxidant, anti-inflammatory, anticancer, antiproliferative, anti-pulmonary fibrosis, hypoglycemic and insecticidal ones, besides being useful in skin care (Hamdan et al., 2016HAMDAN, D.I., MOHAMED, M.E. and EL-SHAZLY, A.M., 2016. Citrus reticulata Blanco cv. Santra leaf and fruit peel: a common waste products, volatile oils composition and biological activities. Journal of Medicinal Plants Research, vol. 10, no. 30, pp. 457-467. http://dx.doi.org/10.5897/JMPR2016.6139.
http://dx.doi.org/10.5897/JMPR2016.6139...
; Apraj and Pandita, 2016APRAJ, V.D. and PANDITA, N.S., 2016. Evaluation of skin anti-aging potential of Citrus reticulata blanco peel. Pharmacognosy Research, vol. 8, no. 3, pp. 160-168. http://dx.doi.org/10.4103/0974-8490.182913. PMid:27365982.
http://dx.doi.org/10.4103/0974-8490.1829...
). Therefore, this study aimed to evaluate the chemical composition, larvicidal and anti-Leishmania amazonensis activities of essential oil from C. reticulata fruit peel (CR-EO). So far, larvicidal activity of CR-EO against third instar A. aegypti larvae has not been investigated in the literature.

2. Material and Methods

2.1. Plant material

Fruits were collected in Rio Verde (17°99.4’63.2”S and 51°05.2’44.6”W), a city located in Goiás state, Brazil, on January 18th, 2019, at 9 a.m. The plant was identified by the botanist Luzia Francisca de Souza and a voucher specimen of Citrus reticulata was deposited in the herbarium in Rio Verde, at the Instituto Federal Goiano (IFGOIANO) under identification number #4488.

2.2. Essential oil extraction

Essential oil (EO) was extracted from C. reticulata (CR-EO) fruit peel by hydrodistillation for 3 h in a Clevenger-type apparatus. Hydrodistillation was performed in triplicate. To this end, fruit peel was divided into three 500-g samples and 500 mL distilled water was added to each sample. After manual collection of EO, remaining traces of water were removed with anhydrous sodium sulfate. Filtration was then carried out. EO was stored in an amber bottle and kept in a refrigerator at 4 °C until analysis (Carneiro et al., 2017CARNEIRO, N.S., ALVES, J.M., ALVES, C.C.F., ESPERANDIM, V.R. and MIRANDA, M.L.D., 2017. Óleo essencial das flores de Eugenia klotzschiana (Myrtaceae): composição química e atividades tripanocida e citotóxica in vitro. Revista Virtual de Química, vol. 9, no. 3, pp. 1381-1392. http://dx.doi.org/10.21577/1984-6835.20170080.
http://dx.doi.org/10.21577/1984-6835.201...
).

2.3. Chemical identification of essential oil from C. reticulata fruit peel (CR-EO)

CR-EO was dissolved in ethyl ether and analyzed by gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC–MS), with the use of Shimadzu QP5000 Plus and GCMS2010 Plus (Shimadzu Corporation, Kyoto, Japan) systems. The temperature of the column in GC-FID was programmed to rise from 60 to 240 °C at 3 °C/min and was held at 240 °C for 5 min; the carrier gas was H2 at a flow rate of 1.0 mL/min. The equipment was set to operate in the injection mode; the injection volume was 0.1 µL (split ratio of 1:10), while injector and detector temperatures were 240 and 280 °C, respectively. Relative concentrations of components were obtained by normalizing peak areas (%). Relative areas consisted of the average of triplicate GC-FID analyses. GC-MS conditions and the identification of essential oils have been previously reported (Lemes et al., 2018LEMES, R.S., ALVES, C.C.F., ESTEVAM, E.B.B., SANTIAGO, M.B., MARTINS, C.H.G., SANTOS, T.C.L., CROTTI, A.E.M. and MIRANDA, M.L.D., 2018. Chemical composition and antibacterial activity of essential oils from Citrus aurantifolia leaves and fruit peel against oral pathogenic bacteria. Anais da Academia Brasileira de Ciências, vol. 90, no. 2, pp. 1285-1292. http://dx.doi.org/10.1590/0001-3765201820170847. PMid:29898096.
http://dx.doi.org/10.1590/0001-376520182...
). Identification of volatile components of essential oil from C. reticulata (Table 1) was based on their retention indices on an Rtx-5MS (30 m × 0.25 mm; 0.250 µm) capillary column under the operating conditions used for GC relative to a homologous series of n-alkanes (C8-C20). Structures were computer-matched with Wiley 7, NIST 08, and FFNSC 1.2, and their fragmentation patterns were compared with literature data (Adams, 2007ADAMS, R.P., 2007. Identification of essential oil components by gas chromatography/quadrupole mass spectroscopy. 4th ed. Carol Stream: Allured Publishing Corporation, 804 p.).

Table 1
Chemical composition of essential oil from Citrus reticulata fruit peel (CR-EO).

2.4. Larvicidal assay

Larvae of A. aegypti were obtained from the Laboratório de Patologia Tropical e Saúde Pública that belongs to the Universidade Federal de Goiás, UFG, Brazil. The larvicidal assay was performed in agreement with a previously reported method (Mesquita et al., 2018MESQUITA, R.S., TADEI, W.P. and BASTOS, A.M.B., 2018. Determination of the larvicidal activity of benzoyl thiosemicarbazone and its Ni(II) complex against Aedes aegypti and Anopheles darlingi larvae in Amazonas, Brazil. Journal of Entomology and Nematology, vol. 10, no. 6, pp. 37-42. https://doi.org/10.5897/JEN2018.0207.
https://doi.org/10.5897/JEN2018.0207...
), as follows: larvae were kept in plastic trays under controlled temperature (26 ± 2 °C) and humidity (70-80%) until they reached the third final instar stage. Afterwards, 10 larvae were transferred to 50-mL plastic cups, each containing 10 mL mineral water and ground fish food (TetraMin Tropical Flakes), followed by the addition of 100 μL solution of CR-EO in dimethyl sulfoxide (DMSO) (25-500 μg/mL). After 24 hours, the number of dead larvae was counted and the lethal percentage was calculated. All experiments were carried out in quintuplicate, including a negative control treatment with DMSO, mineral water, larvae and ground fish food. Permethrin was used as a positive control. Larvicidal activities were reported as lethal concentration at 50% (LC50), representing the concentration in micrograms per milliliter that caused 50% larval mortality, with 95% confidence interval. Mortality data were assessed by probit analysis (Finney, 1971FINNEY, D.J., 1971. Probit analysis. Cambridge: Cambridge University Press.). Mortality data were treated by the Polo plus® software (Robertson et al., 2003ROBERTSON, J.L., PREISLER, H.K. and RUSSELL, R.M., 2003. Poloplus probit and logit analysis. Berkeley: Leora Software, pp. 1-36.) with 95% confidence interval and values of P < 0.05 were considered statistically significant.

2.5. Antileishmanial assay

To evaluate antileishmanial activity, L. amazonensis promastigote forms (MHOM/BR/PH8) were maintained in RPMI 1640 (Gibco) culture medium supplemented with 10% fetal bovine serum, penicillin (100 UI/mL) and streptomycin (100 μg/mL). Subsequently, about 1 x 106 parasites were distributed on 96-well plates, while CR-EO previously dissolved in 100% dimethylsulfoxide (DMSO, stock solution 100 mM) (Synth) was added to the cultures at concentrations of 6.25 to 100 μg/mL. Amphotericin B (Sigma Aldrich, 97% purity), at concentrations ranging from 0.011 to 0.19 μg/mL, was added to cultures and used as positive control. Cultures were incubated in a BOD (Quimis) incubator at 25 °C for 24 h and antileishmanial activity was determined by verifying whether growth of promastigote forms was inhibited. It was carried out by counting the total number of live promastigotes in the Neubauer (Global Glass - Porto Alegre, BR) chamber on the basis of flagellar motility. RPMI 1640 medium (Gibco) containing 0.1% DMSO (Synth) (highest concentration) was used. Results were expressed as the average of the percentage of growth inhibition relative to the negative control (0.1% DMSO) (Cabral et al., 2020CABRAL, F.D., FERNANDES, C.C., RIBEIRO, A.B., SQUARISI, I.S., TAVARES, D.C., CANDIDO, A.C.B.B., MAGALHÃES, L.G., SOUZA, J.M., MARTINS, C.H.G. and MIRANDA, M.L.D., 2020. Bioactivities of essential oils from different parts of Spiranthera odoratissima (Rutaceae). Rodriguésia, vol. 71, e00902019. http://dx.doi.org/10.1590/2175-7860202071050.
http://dx.doi.org/10.1590/2175-786020207...
).

3. Results and Discussion

Volatile compounds were identified by gas chromatography-flame ionization detection (GC-FID) and gas chromatography–mass spectrometry (GC-MS). Thirteen compounds, which represent 97.8% of all components, were identified in the oil from fruit peel. Major compounds found in CR-EO were limonene (85.7%), ɣ-terpinene (6.7%) and myrcene (2.1%) (Table 1).

Previous reports of EOs from fruit peel from other C. reticulata specimens showed that terpenes limonene, sabinene, linalool, ɣ-terpinene, octanal and capraldehyde were its major constituents (Hamdan et al., 2016HAMDAN, D.I., MOHAMED, M.E. and EL-SHAZLY, A.M., 2016. Citrus reticulata Blanco cv. Santra leaf and fruit peel: a common waste products, volatile oils composition and biological activities. Journal of Medicinal Plants Research, vol. 10, no. 30, pp. 457-467. http://dx.doi.org/10.5897/JMPR2016.6139.
http://dx.doi.org/10.5897/JMPR2016.6139...
; Martins et al., 2017MARTINS, M.H.G., FRACAROLLI, L., VIEIRA, T.M., DIAS, H.J., CRUZ, M.G., DEUS, C.C.H., NICOLELLA, H.D., STEFANI, R., RODRIGUES, V., TAVARES, D.C., MAGALHÃES, L.G. and CROTTI, A.E.M., 2017. Schistosomicidal effects of the essential oils of Citrus limonia and Citrus reticulata against Schistosoma mansoni. Chemistry & Biodiversity, vol. 14, no. 1, pp. e1600194. http://dx.doi.org/10.1002/cbdv.201600194. PMid:27936310.
http://dx.doi.org/10.1002/cbdv.201600194...
). EOs from different Citrus species are chemically similar with the predominance of the monoterpene limonene, characteristics that are pointed out by Bozkurt et al. (2017)BOZKURT, T., GULNAZ, O. and KAÇAR, Y.A., 2017. Chemical composition of the essential oils from some citrus species and evaluation of the antimicrobial activity. Journal of Environmental Science. Toxicology and Food Technology, vol. 11, pp. 29-33. http://dx.doi.org/10.9790/2402-1110032933.
http://dx.doi.org/10.9790/2402-111003293...
. High limonene concentration (74.38%) was also identified in C. reticulata peel grown in Spain (Espina et al., 2011ESPINA, L., SOMOLINOS, M., LORÁN, S., CONCHELLO, P., GARCÍA, D. and PAGÁN, R., 2011. Chemical composition of commercial citrus fruit essential oils and evaluation of their antimicrobial activity acting alone or in combined process. Food Control, vol. 22, no. 6, pp. 896-902. http://dx.doi.org/10.1016/j.foodcont.2010.11.021.
http://dx.doi.org/10.1016/j.foodcont.201...
). A recent study of ten Citrus species should be highlighted, since it reinforces limonene and aldehyde compounds in CR-EO (González-Mas et al., 2019GONZÁLEZ-MAS, M.C., RAMBLA, J.L., LÓPEZ-GRESA, M.P., BLÁZQUEZ, M.A. and GRANELL, A., 2019. Volatile compounds in Citrus essential oils: a comprehensive review. Frontiers in Plant Science, vol. 10, pp. 12. http://dx.doi.org/10.3389/fpls.2019.00012. PMid:30804951.
http://dx.doi.org/10.3389/fpls.2019.0001...
).

CR-EO had its larvicidal activity investigated against third instar A. aegypti larvae. Initially, larvae in contact with CR-EO showed accelerated movement. However, after prolonged exposure, they began to exhibit tremor and slow or lethargic movement, even when artificially stimulated. CR-EO also caused darkening of the entire larval body. Doses of 12.5, 25, 50 and 100 µg/mL resulted in 10.2, 25.1, 51.7 and 85.3% of dead larvae, respectively, while 150 µg/mL ensured 100% mortality. LC50 of CR-EO was 58.35 µg/mL. Even though EOs from several Citrus species have already been tested against A. aegypti larvae, this is the first promising report of C. reticulata. Larvicidal and insecticidal activities have already been evaluated in EOs from C. sinensis, C. limon, C. grandis, C. aurantifolia, C. hystrix, C. maxima and C. medica (Araújo et al., 2016ARAÚJO, A.F.O., RIBEIRO-PAES, J.T., DEUS, J.T., CAVALCANTI, S.C.H., NUNES, R.S., ALVES, P.B. and MACORIS, M.L.G., 2016. Larvicidal activity of Syzygium aromaticum (L.) Merr and Citrus sinensis (L.) Osbeck essential oil and their antagonistic effects with temephos in resistant populations of Aedes aegypti. Memórias do Instituto Oswaldo Cruz, vol. 111, no. 7, pp. 443-449. http://dx.doi.org/10.1590/0074-02760160075. PMid:27384083.
http://dx.doi.org/10.1590/0074-027601600...
; Gomes et al., 2019GOMES, P.R.B., OLIVEIRA, M.B., SOUSA, D.A., SILVA, J.C., FERNANDES, R.P., LOUZEIRO, H.C., OLIVEIRA, R.W.S., PAULA, M.L., FILHO, V.E.M. and FONTENELE, M.A., 2019. Larvicidal activity, molluscicide and toxicity of the essential oil of Citrus limon peels against, respectively, Aedes aegypti, Biomphalaria glabrata and Artemia salina. Eclética Química Journal, vol. 44, no. 4, pp. 85-95. http://dx.doi.org/10.26850/1678-4618eqj.v44.4.2019.p85-95.
http://dx.doi.org/10.26850/1678-4618eqj....
; Sarma et al., 2017SARMA, R., KHANIKOR, B. and MAHANTA, S., 2017. Essential oil from Citrus grandis (Sapindales: Rutaceae) as insecticide against Aedes aegypti (L) (Diptera: Culicidae). Intertnational Journal of Mosquito Research, vol. 4, pp. 88-92., 2019SARMA, R., ADHIKARI, K., MAHANTA, S. and KHANIKOR, B., 2019. Insecticidal activities of Citrus aurantifolia essential oil against Aedes aegypti (Diptera: culicidae). Toxicology Reports, vol. 6, pp. 1091-1096. http://dx.doi.org/10.1016/j.toxrep.2019.10.009. PMid:31687359.
http://dx.doi.org/10.1016/j.toxrep.2019....
; Soonwera, 2015SOONWERA, M., 2015. Efficacy of essential oils from Citrus plantas against mosquito vectors Aedes aegypti (Linn.) and Culex quinquefasciatus (Say). Agricultural Technology, vol. 11, pp. 669-681.). High larvicidal activity could be due to the high limonene concentration (85.7%), a botanical insecticide which has been patented as an active agent of larvicidal formulations (Dias and Moraes, 2014DIAS, C.N. and MORAES, D.F.C., 2014. Essential oils and their compounds as Aedes aegypti L. (Diptera: Culicidae) larvicides. Parasitology Research, vol. 113, no. 2, pp. 565-592. http://dx.doi.org/10.1007/s00436-013-3687-6. PMid:24265058.
http://dx.doi.org/10.1007/s00436-013-368...
).

Regarding antileishmanial activity, CR-EO was promising against promastigote forms of L. amazonensis (IC50 = 8.23 µg/mL) (Table 2).

Table 2
Antileishmanial activity of essential oil from Citrus reticulata fruit peel (CR-EO).

Several authors have reported that EOs whose values are IC50 < 10 µg/mL are highly active (Silva et al., 2020SILVA, F.F.A., FERNANDES, C.C., OLIVEIRA, G.A., CANDIDO, A.C.B.B., MAGALHÃES, L.G., VIEIRA, T.M., CROTTI, A.E.M., SILVA, C.A. and MIRANDA, M.L.D., 2020. In vitro antileishmanial and antioxidant activities of essential oils from different parts of Murraya paniculata (L.) Jack: a species of Rutaceae that occur in the Cerrado bioma in Brazil. Australian Journal of Crop Science, vol. 14, no. 2, pp. 347-353. http://dx.doi.org/10.21475/ajcs.20.14.02.p1966.
http://dx.doi.org/10.21475/ajcs.20.14.02...
; Almeida et al., 2020ALMEIDA, K.C.R., SILVA, B.B., ALVES, C.C.F., VIEIRA, T.M., CROTTI, A.E.M., SOUZA, J.M., MARTINS, C.H.G., RIBEIRO, A.B., SQUARISI, I.S., TAVARES, D.C., BERNABÉ, L.S., MAGALHÃES, L.G. and MIRANDA, M.L.D., 2020. Biological properties and chemical composition of essential oil from Nectandra megapotamica (Spreng.) Mez. leaves (Lauraceae). Natural Product Research, vol. 34, no. 21, pp. 3149-3153. http://dx.doi.org/10.1080/14786419.2019.1608539. PMid:31084218.
http://dx.doi.org/10.1080/14786419.2019....
). The range of IC50 values that considers EOs highly active has been described by Estevam et al. (2016)ESTEVAM, E.B.B., MIRANDA, M.L.D., ALVES, J.M., EGEA, M.B., PEREIRA, P.S., MARTINS, C.H.G., ESPERANDIM, V.R., MAGALHÃES, L.G., BOLELA, A.C., CAZAL, C.M., SOUZA, A.F. and ALVES, C.C.F., 2016. Composição química e atividades biológicas dos óleos essenciais das folhas frescas de Citrus limonia Osbeck e Citrus latifolia Tanaka (Rutaceae). Revista Virtual de Química, vol. 8, pp. 1842-1854. http://dx.doi.org/10.21577/1984-6835.20160124.
http://dx.doi.org/10.21577/1984-6835.201...
, who studied EOs from C. limonia and C. latifolia fresh leaves. It should be highlighted that results found by the study reported by this paper do not agree with the ones reported by Monzote et al. (2019)MONZOTE, L., HERRERA, I., SATYAL, P. and SETZER, W.N., 2019. In-vitro evaluation of 52 commercially-available essential oils against Leishmania amazonensis. Molecules, vol. 24, no. 7, pp. 1248. http://dx.doi.org/10.3390/molecules24071248. PMid:30934998.
http://dx.doi.org/10.3390/molecules24071...
, since the IC50 value of commercially obtained CR-EO was 70.7 µg/mL. This fact leads to questioning the nature of EOs that are commercialized with no compliance with regulations. On the other hand, high antileishmanial activity of CR-EO found by this study may also be justified by the amount of limonene, a monoterpene that exhibits this activity against parasites that belong to the genus Leishmania (Arruda et al., 2009ARRUDA, D.C., MIGUEL, D.C., YOKOYAMA-YASUNAKA, J.K.U., KATZIN, A.M. and ULIANA, S.R.B., 2009. Inhibitory activity of limonene against Leishmania parasites in vitro and in vivo. Biomedicine and Pharmacotherapy, vol. 63, no. 9, pp. 643-649. http://dx.doi.org/10.1016/j.biopha.2009.02.004. PMid:19321295.
http://dx.doi.org/10.1016/j.biopha.2009....
).

4. Conclusion

This study described the chemical composition of CR-EO, in which thirteen compounds were identified, and showed their high larvicidal activity against Aedes aegypti larvae and anti-Leishmania amazonensis promastigote forms. Limonene, as the major compound in CR-EO, could explain their high larvicidal and antileishmanial activities. This communication is the first report of larvicidal evaluation of CR-EO.

Acknowledgements

The authors are grateful to FAPEG, CNPq, CAPES and IF GOIANO – Campus Rio Verde for their financial support.

References

  • ADAMS, R.P., 2007. Identification of essential oil components by gas chromatography/quadrupole mass spectroscopy 4th ed. Carol Stream: Allured Publishing Corporation, 804 p.
  • ALMEIDA, K.C.R., SILVA, B.B., ALVES, C.C.F., VIEIRA, T.M., CROTTI, A.E.M., SOUZA, J.M., MARTINS, C.H.G., RIBEIRO, A.B., SQUARISI, I.S., TAVARES, D.C., BERNABÉ, L.S., MAGALHÃES, L.G. and MIRANDA, M.L.D., 2020. Biological properties and chemical composition of essential oil from Nectandra megapotamica (Spreng.) Mez. leaves (Lauraceae). Natural Product Research, vol. 34, no. 21, pp. 3149-3153. http://dx.doi.org/10.1080/14786419.2019.1608539 PMid:31084218.
    » http://dx.doi.org/10.1080/14786419.2019.1608539
  • APRAJ, V.D. and PANDITA, N.S., 2016. Evaluation of skin anti-aging potential of Citrus reticulata blanco peel. Pharmacognosy Research, vol. 8, no. 3, pp. 160-168. http://dx.doi.org/10.4103/0974-8490.182913 PMid:27365982.
    » http://dx.doi.org/10.4103/0974-8490.182913
  • ARAÚJO, A.F.O., RIBEIRO-PAES, J.T., DEUS, J.T., CAVALCANTI, S.C.H., NUNES, R.S., ALVES, P.B. and MACORIS, M.L.G., 2016. Larvicidal activity of Syzygium aromaticum (L.) Merr and Citrus sinensis (L.) Osbeck essential oil and their antagonistic effects with temephos in resistant populations of Aedes aegypti. Memórias do Instituto Oswaldo Cruz, vol. 111, no. 7, pp. 443-449. http://dx.doi.org/10.1590/0074-02760160075 PMid:27384083.
    » http://dx.doi.org/10.1590/0074-02760160075
  • ARRUDA, D.C., MIGUEL, D.C., YOKOYAMA-YASUNAKA, J.K.U., KATZIN, A.M. and ULIANA, S.R.B., 2009. Inhibitory activity of limonene against Leishmania parasites in vitro and in vivo. Biomedicine and Pharmacotherapy, vol. 63, no. 9, pp. 643-649. http://dx.doi.org/10.1016/j.biopha.2009.02.004 PMid:19321295.
    » http://dx.doi.org/10.1016/j.biopha.2009.02.004
  • BOZKURT, T., GULNAZ, O. and KAÇAR, Y.A., 2017. Chemical composition of the essential oils from some citrus species and evaluation of the antimicrobial activity. Journal of Environmental Science. Toxicology and Food Technology, vol. 11, pp. 29-33. http://dx.doi.org/10.9790/2402-1110032933
    » http://dx.doi.org/10.9790/2402-1110032933
  • BRADY, O.J., GETHING, P.W., BHATT, S., MESSINA, J.P., BROWNSTEIN, J.S., HOEN, A.G., MOYES, C.L., FARLOW, A.W., SCOTT, T.W. and HAY, S.I., 2012. Refining the global spatial limits of dengue virus transmission by evidence-based consensus. PLoS Neglected Tropical Diseases, vol. 6, no. 8, e1760. http://dx.doi.org/10.1371/journal.pntd.0001760 PMid:22880140.
    » http://dx.doi.org/10.1371/journal.pntd.0001760
  • CABRAL, F.D., FERNANDES, C.C., RIBEIRO, A.B., SQUARISI, I.S., TAVARES, D.C., CANDIDO, A.C.B.B., MAGALHÃES, L.G., SOUZA, J.M., MARTINS, C.H.G. and MIRANDA, M.L.D., 2020. Bioactivities of essential oils from different parts of Spiranthera odoratissima (Rutaceae). Rodriguésia, vol. 71, e00902019. http://dx.doi.org/10.1590/2175-7860202071050
    » http://dx.doi.org/10.1590/2175-7860202071050
  • CARNEIRO, N.S., ALVES, J.M., ALVES, C.C.F., ESPERANDIM, V.R. and MIRANDA, M.L.D., 2017. Óleo essencial das flores de Eugenia klotzschiana (Myrtaceae): composição química e atividades tripanocida e citotóxica in vitro. Revista Virtual de Química, vol. 9, no. 3, pp. 1381-1392. http://dx.doi.org/10.21577/1984-6835.20170080
    » http://dx.doi.org/10.21577/1984-6835.20170080
  • DIAS, C.N. and MORAES, D.F.C., 2014. Essential oils and their compounds as Aedes aegypti L. (Diptera: Culicidae) larvicides. Parasitology Research, vol. 113, no. 2, pp. 565-592. http://dx.doi.org/10.1007/s00436-013-3687-6 PMid:24265058.
    » http://dx.doi.org/10.1007/s00436-013-3687-6
  • DOSOKY, N.S. and SETZER, W.N., 2018. Biological activities and safety of Citrus spp. Essential oils. International Journal of Molecular Sciences, vol. 19, no. 7, pp. 1966. http://dx.doi.org/10.3390/ijms19071966 PMid:29976894.
    » http://dx.doi.org/10.3390/ijms19071966
  • ESPINA, L., SOMOLINOS, M., LORÁN, S., CONCHELLO, P., GARCÍA, D. and PAGÁN, R., 2011. Chemical composition of commercial citrus fruit essential oils and evaluation of their antimicrobial activity acting alone or in combined process. Food Control, vol. 22, no. 6, pp. 896-902. http://dx.doi.org/10.1016/j.foodcont.2010.11.021
    » http://dx.doi.org/10.1016/j.foodcont.2010.11.021
  • ESTEVAM, E.B.B., MIRANDA, M.L.D., ALVES, J.M., EGEA, M.B., PEREIRA, P.S., MARTINS, C.H.G., ESPERANDIM, V.R., MAGALHÃES, L.G., BOLELA, A.C., CAZAL, C.M., SOUZA, A.F. and ALVES, C.C.F., 2016. Composição química e atividades biológicas dos óleos essenciais das folhas frescas de Citrus limonia Osbeck e Citrus latifolia Tanaka (Rutaceae). Revista Virtual de Química, vol. 8, pp. 1842-1854. http://dx.doi.org/10.21577/1984-6835.20160124
    » http://dx.doi.org/10.21577/1984-6835.20160124
  • FINNEY, D.J., 1971. Probit analysis Cambridge: Cambridge University Press.
  • GOMES, P.R.B., OLIVEIRA, M.B., SOUSA, D.A., SILVA, J.C., FERNANDES, R.P., LOUZEIRO, H.C., OLIVEIRA, R.W.S., PAULA, M.L., FILHO, V.E.M. and FONTENELE, M.A., 2019. Larvicidal activity, molluscicide and toxicity of the essential oil of Citrus limon peels against, respectively, Aedes aegypti, Biomphalaria glabrata and Artemia salina. Eclética Química Journal, vol. 44, no. 4, pp. 85-95. http://dx.doi.org/10.26850/1678-4618eqj.v44.4.2019.p85-95
    » http://dx.doi.org/10.26850/1678-4618eqj.v44.4.2019.p85-95
  • GONZÁLEZ-MAS, M.C., RAMBLA, J.L., LÓPEZ-GRESA, M.P., BLÁZQUEZ, M.A. and GRANELL, A., 2019. Volatile compounds in Citrus essential oils: a comprehensive review. Frontiers in Plant Science, vol. 10, pp. 12. http://dx.doi.org/10.3389/fpls.2019.00012 PMid:30804951.
    » http://dx.doi.org/10.3389/fpls.2019.00012
  • HAMDAN, D.I., MOHAMED, M.E. and EL-SHAZLY, A.M., 2016. Citrus reticulata Blanco cv. Santra leaf and fruit peel: a common waste products, volatile oils composition and biological activities. Journal of Medicinal Plants Research, vol. 10, no. 30, pp. 457-467. http://dx.doi.org/10.5897/JMPR2016.6139
    » http://dx.doi.org/10.5897/JMPR2016.6139
  • LEMES, R.S., ALVES, C.C.F., ESTEVAM, E.B.B., SANTIAGO, M.B., MARTINS, C.H.G., SANTOS, T.C.L., CROTTI, A.E.M. and MIRANDA, M.L.D., 2018. Chemical composition and antibacterial activity of essential oils from Citrus aurantifolia leaves and fruit peel against oral pathogenic bacteria. Anais da Academia Brasileira de Ciências, vol. 90, no. 2, pp. 1285-1292. http://dx.doi.org/10.1590/0001-3765201820170847 PMid:29898096.
    » http://dx.doi.org/10.1590/0001-3765201820170847
  • MARTINS, M.H.G., FRACAROLLI, L., VIEIRA, T.M., DIAS, H.J., CRUZ, M.G., DEUS, C.C.H., NICOLELLA, H.D., STEFANI, R., RODRIGUES, V., TAVARES, D.C., MAGALHÃES, L.G. and CROTTI, A.E.M., 2017. Schistosomicidal effects of the essential oils of Citrus limonia and Citrus reticulata against Schistosoma mansoni. Chemistry & Biodiversity, vol. 14, no. 1, pp. e1600194. http://dx.doi.org/10.1002/cbdv.201600194 PMid:27936310.
    » http://dx.doi.org/10.1002/cbdv.201600194
  • MESQUITA, R.S., TADEI, W.P. and BASTOS, A.M.B., 2018. Determination of the larvicidal activity of benzoyl thiosemicarbazone and its Ni(II) complex against Aedes aegypti and Anopheles darlingi larvae in Amazonas, Brazil. Journal of Entomology and Nematology, vol. 10, no. 6, pp. 37-42. https://doi.org/10.5897/JEN2018.0207
    » https://doi.org/10.5897/JEN2018.0207
  • MONZOTE, L., HERRERA, I., SATYAL, P. and SETZER, W.N., 2019. In-vitro evaluation of 52 commercially-available essential oils against Leishmania amazonensis. Molecules, vol. 24, no. 7, pp. 1248. http://dx.doi.org/10.3390/molecules24071248 PMid:30934998.
    » http://dx.doi.org/10.3390/molecules24071248
  • MOREIRA, R.R.D., SANTOS, A.G., CARVALHO, F.A., PEREGO, C.H., CREVELIN, E.J., CROTTI, A.E.M., COGO, J., CARDOSO, M.L.C. and NAKAMURA, C.V., 2019. Antileishmanial activity of Melampodium divaricatum and Casearia sylvestris essential oils on Leishmania amazonensis. Revista do Instituto de Medicina Tropical de São Paulo, vol. 61, pp. e33. http://dx.doi.org/10.1590/s1678-9946201961033 PMid:31269109.
    » http://dx.doi.org/10.1590/s1678-9946201961033
  • NEVES, I.A., REZENDE, S.R.F., KIRK, J.M., PONTES, E.G. and CARVALHO, M.G., 2017. Composition and larvicidal activity of essential oil of Eugenia candolleana DC. (Myrtaceae) against Aedes aegypti. Revista Virtual de Química, vol. 9, no. 6, pp. 2305-2315. http://dx.doi.org/10.21577/1984-6835.20170138
    » http://dx.doi.org/10.21577/1984-6835.20170138
  • ROBERTSON, J.L., PREISLER, H.K. and RUSSELL, R.M., 2003. Poloplus probit and logit analysis Berkeley: Leora Software, pp. 1-36.
  • SARMA, R., ADHIKARI, K., MAHANTA, S. and KHANIKOR, B., 2019. Insecticidal activities of Citrus aurantifolia essential oil against Aedes aegypti (Diptera: culicidae). Toxicology Reports, vol. 6, pp. 1091-1096. http://dx.doi.org/10.1016/j.toxrep.2019.10.009 PMid:31687359.
    » http://dx.doi.org/10.1016/j.toxrep.2019.10.009
  • SARMA, R., KHANIKOR, B. and MAHANTA, S., 2017. Essential oil from Citrus grandis (Sapindales: Rutaceae) as insecticide against Aedes aegypti (L) (Diptera: Culicidae). Intertnational Journal of Mosquito Research, vol. 4, pp. 88-92.
  • SILVA, F.F.A., FERNANDES, C.C., OLIVEIRA, G.A., CANDIDO, A.C.B.B., MAGALHÃES, L.G., VIEIRA, T.M., CROTTI, A.E.M., SILVA, C.A. and MIRANDA, M.L.D., 2020. In vitro antileishmanial and antioxidant activities of essential oils from different parts of Murraya paniculata (L.) Jack: a species of Rutaceae that occur in the Cerrado bioma in Brazil. Australian Journal of Crop Science, vol. 14, no. 2, pp. 347-353. http://dx.doi.org/10.21475/ajcs.20.14.02.p1966
    » http://dx.doi.org/10.21475/ajcs.20.14.02.p1966
  • SOONWERA, M., 2015. Efficacy of essential oils from Citrus plantas against mosquito vectors Aedes aegypti (Linn.) and Culex quinquefasciatus (Say). Agricultural Technology, vol. 11, pp. 669-681.
  • WANKHAR, W., SRINIVASAN, S. and RATHINASAMY, S., 2015. HPTLC analysis of Scoparia dulcis Linn (Scrophulariaceae) and its larvicidal potential against dengue vector Aedes aegypti. Natural Product Research, vol. 29, no. 18, pp. 1757-1760. http://dx.doi.org/10.1080/14786419.2014.999060 PMid:25573588.
    » http://dx.doi.org/10.1080/14786419.2014.999060

Publication Dates

  • Publication in this collection
    28 June 2021
  • Date of issue
    2023

History

  • Received
    13 Jan 2021
  • Accepted
    03 Mar 2021
Creative Common - by 4.0
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.
Instituto Internacional de Ecologia R. Bento Carlos, 750, 13560-660 São Carlos SP - Brasil, Tel. e Fax: (55 16) 3362-5400 - São Carlos - SP - Brazil
E-mail: bjb@bjb.com.br