Abstracts
Phytotherapy can be an alternative for the control of gastrointestinal parasites of small ruminants. This study evaluated the efficacy of Alpinia zerumbet, Coriandrum sativum, Tagetes minuta and Lantana camara essential oils by two in vitro assays on Haemonchus contortus, an egg hatch test (EHT) and larval development test (LDT). No effect was observed for L. camara in the EHT. A. zerumbet, C. sativum and T. minuta essential oils exhibited a dose-dependent effect in the EHT, inhibiting 81.2, 99 and 98.1% of H. contortus larvae hatching, respectively, at a concentration of 2.5 mg mL-1. The effective concentration to inhibit 50% (EC50) of egg hatching was 0.94, 0.63 and 0.53 mg mL-1 for A. zerumbet, C. sativum and T. minuta essential oils, respectively. In LDT, L. camara, A. zerumbet, C. sativum and T. minuta at concentration of 10 mg mL-1 inhibited 54.9, 94.2, 97.8 and 99.5% of H. contortus larval development, presenting EC50 values of 6.32, 3.88, 2.89 and 1.67 mg mL-1, respectively. Based on the promising results presented in this in vitro model, it may be possible use of these essential oils to control gastrointestinal nematodes. However, their anthelmintic activity should be confirmed in vivo.
Phytotherapy; anthelmintic; gastrointestinal nematodes
Fitoterapia pode ser uma alternativa para o controle de parasitas gastrintestinais de pequenos ruminantes. Este estudo avaliou a eficácia dos óleos essenciais de Alpinia zerumbet, Coriandrum sativum, Tagetes minuta e Lantana camara sobre Haemonchus contortus através de dois testes in vitro, teste de eclosão dos ovos (TEO) e teste de desenvolvimento larval (TDL). Nenhum efeito foi observado para L. camara no TEO. Os óleos essenciais de A. zerumbet, C. sativum e T. minuta exibiram um efeito dose dependente no TEO inibindo a eclosão das larvas de H. contortus em 81,2, 99 e 98,1%, respectivamente, na concentração de 2,5 mg mL-1. A concentração efetiva para inibir 50% (CE50) da eclosão dos ovos foi de 0,94, 0,63 e 0,53 mg mL-1 para os óleos essenciais de A. zerumbet, C. sativum e T. minuta, respectivamente. No TDL, 10 mg mL-1 de L. camara, A. zerumbet, C. sativum e T. minuta inibiram em 54,9, 94,2, 97,8 e 99,5% do desenvolvimento larvar, apresentando valores de CE50 de 6,32, 3,88, 2,89 e 1,67 mg mL-1, respectivamente. Com base nos resultados promissores apresentados neste modelo in vitro, pode ser possível a utilização destes óleos essenciais para controlar os nematoides gastrintestinais. No entanto, a sua atividade anti-helmíntica deve ser confirmada in vivo.
Fitoterapia; anti-helmínticos; nematoides gastrintestinais
Introduction
Parasitic nematodes are a major threat to livestock production worldwide
leading to huge economic losses (ADEMOLA; ELLOF, 2010), because of the support
treatments and increased manpower that the infected animals require and because of
the high mortality rate within flocks, especially in small ruminants (DIEHL et al., 2004Diehl MS, Atindehou KK, Tere H, Betschart B. Prospect for
anthelminthic plants in the Ivory Coast using ethnobotanical criteria. J
Ethnopharmacol 2004; 95(2-3): 277-284.
http://dx.doi.org/10.1016/j.jep.2004.07.012
http://dx.doi.org/10.1016/j.jep.2004.07....
). Anthelmintics have been
used to minimize the losses caused by helminth infections. However, the efficacy of
anthelmintics is increasingly endangered by development of resistance in nematode
populations (MILLER et al., 2012Miller CM, Waghorn TS, Leathwick DM, Candy PM, Oliver AMB, Watson
TG. The production cost of anthelmintic resistance in lambs. Vet Parasitol 2012;
186(3-4): 376-381.
http://dx.doi.org/10.1016/j.vetpar.2011.11.063
http://dx.doi.org/10.1016/j.vetpar.2011....
). Control by
anthelmintics has presented problems, such as the development of resistance in
nematodes, the poor availability and high cost of commercial products, especially to
low income farmers in developing countries, and increasing concern over the risk of
environmental contamination because the presence of drug residues in animal products
when pure compounds are administered (WALLER,
2006Waller PJ. From discovery to development: Current industry
perspectives for the development of novel methods of helminth control in
livestock. Vet Parasitol 2006; 139(1-3): 1-14.
http://dx.doi.org/10.1016/j.vetpar.2006.02.036
http://dx.doi.org/10.1016/j.vetpar.2006....
). These concerns have led to the search for and evaluation of
alternative control methods (ATHANASIADOU et al.,
2001Athanasiadou S, Kyriazakis I, Jackson F, Coop RL. Direct
anthelmintic effects of condensed tannins towards different gastrointestinal
nematodes of sheep: in vitro and in vivo studies. Vet Parasitol 2001; 99(3):
205-219. http://dx.doi.org/10.1016/S0304-4017(01)00467-8
http://dx.doi.org/10.1016/S0304-4017(01)...
). One promising alternative investigated for the control of
gastrointestinal parasites has been the use of phytotherapics (MAPHOSA et al., 2010Maphosa V, Masika PJ, Bizimenyera ES, Eloff JN. In vitro
anthelminthic activity of crude aqueous extracts of Aloe ferox, Leonotis
leonurus and Elephantorrhiza elephantina against Haemonchus contortus. Trop Anim
Health Prod 2010; 42(2): 301-307.
http://dx.doi.org/10.1007/s11250-009-9421-9
http://dx.doi.org/10.1007/s11250-009-942...
). Efforts are being made to evaluate
several medicinal plants for their anthelmintic potential in different parts of the
world. Haemonchus contortus, a highly pathogenic and one of the
most prevalent nematode parasite, has been used by researchers to evaluate the
anthelmintic effects of plants in small ruminants (EGUALE et al., 2011Eguale T, Tadesse D, Giday M. In vitro anthelmintic activity of
crude extracts of five medicinal plants against egg-hatching and larval
development of Haemonchus contortus. J Ethnopharmacol 2011; 137(1): 108-113.
http://dx.doi.org/10.1016/j.jep.2011.04.063
http://dx.doi.org/10.1016/j.jep.2011.04....
).
Among the criteria used to select plant species are their availability and
the existence of studies researching their biological activity on nematodes. Of the
plant species that have been documented as having medicinal effects against
helminths, Lantana camara, Tagetes minuta (ALBUQUERQUE et al., 2007Albuquerque UP, Medeiros PM, Almeida AL, Monteiro JM, Lins EMF
N°, Melo JG, et al. Medicinal plants of the caatinga (semi-arid) vegetation
of NE Brazil: A quantitative approach. J Ethnopharmacol 2007; 114(3): 325-354.
http://dx.doi.org/10.1016/j.jep.2007.08.017
http://dx.doi.org/10.1016/j.jep.2007.08....
), Alpinia zerumbet
(ALMEIDA, 1993) and Coriandrum sativum (HUSSAIN et al., 2008Hussain A, Khan MN, Iqbal Z, Sajid MS. An account of the botanical
anthelmintics used in traditional veterinary practices in Sahiwal district of
Punjab, Pakistan. J Ethnopharmacol 2008; 119(1): 185-190.
http://dx.doi.org/10.1016/j.jep.2008.06.034
http://dx.doi.org/10.1016/j.jep.2008.06....
) are commonly cited.
L. camara, belonging to the Verbenaceae family, exhibits
antimicrobial activity (DEENA; THOPPIL,
2000Deena MJ, Thoppil JE. Antimicrobial activity of the essential oil of
Lantana camara. Fitoterapia 2000; 71(4): 453-455.
http://dx.doi.org/10.1016/S0367-326X(00)00140-4
http://dx.doi.org/10.1016/S0367-326X(00)...
), efficacy against termite workers (VERMA; VERMA, 2006Verma RK, Verma SK. Phytochemical and termiticidal study of Lantana
camara var. aculeata leaves. Fitoterapia 2006; 77(6): 466-468.
http://dx.doi.org/10.1016/j.fitote.2006.05.014
http://dx.doi.org/10.1016/j.fitote.2006....
) and has a nematostatic effect on the phytonematode
Meloidogyne incognita (DIAS et
al., 2000Dias CR, Schwan AV, Ezequiel DP, Sarmento MC, Ferraz S. Efeito de
extratos aquosos de plantas medicinais na sobrevivência de Junenis de
Meloidogyne incognita. Nematol Bras 2000; 24(2): 203-210.). T. minuta, from the Asteraceae family, has
fungitoxic effects (ROZWALKA et al., 2008Rozwalka LC, Lima MLRZC, Mio LLM, Nakashima T. Extratos, decoctos e
óleos essenciais de plantas medicinais e aromáticas na inibição de Glomerella
cingulata e Colletotrichum gloeosporioides de frutos de goiaba. Cienc Rural
2008; 38(2): 301-307.
http://dx.doi.org/10.1590/S0103-84782008000200001
http://dx.doi.org/10.1590/S0103-84782008...
),
activity against Anopheles stephensi larvae (HADJIAKHOONDI et al., 2005Hadjiakhoondi A, Vatandoost H, Khanavi M, Abaee RM, Karami M.
Biochemical investigation of different extracts and larvicidal activity of
Tagetes minuta L. on Anopheles stephensi larvae. Iran J Pharm Sci 2005; 1(2):
81-84.), antibacterial activity (SOUZA et al., 2000Souza CAS, Avancini CAM, Wiest JM. Atividade antimicrobiana de
Tagetes minuta L. - Compositae (Chinchilho) frente a bactérias Gram-positivas e
Gram-negativas. Braz J Vet Res Anim Sci 2000; 37(6): 429-433.
http://dx.doi.org/10.1590/S1413-95962000000600001
http://dx.doi.org/10.1590/S1413-95962000...
) and effect on H.
contortus egg and larvae (MACEDO et
al., 2012Macedo ITF, Bevilaqua CML, Oliveira LMB, Camurça-Vasconcelos ALF,
Morais SM, Machado LKA, et al. In vitro activity of Lantana camara, Alpinia
zerumbet, Mentha villosa and Tagetes minuta decoctions on Haemonchus contortus
eggs and larvae. Vet Parasitol 2012; 190(3-4): 504-509.
http://dx.doi.org/10.1016/j.vetpar.2012.07.001
http://dx.doi.org/10.1016/j.vetpar.2012....
). A. zerumbet, from the Zingiberaceae family,
exhibits antioxidant (ELZAAWELY et al.,
2007Elzaawely AA, Xuan TD, Tawata S. Essential oils, kava pyrones and
phenolic compounds from leaves and rhizomes of Alpinia zerumbet (Pers.) B.L.
Burtt. & R.M. Sm. and their antioxidant activity. Food Chem 2007; 103(2):
486-494. http://dx.doi.org/10.1016/j.foodchem.2006.08.025
http://dx.doi.org/10.1016/j.foodchem.200...
), antibacterial and antifungal activities (VICTóRIO et al., 2009Victório CP, Alviano DS, Alviano CS, Lage CLS. Chemical composition
of the fractions of leaf oil of Alpinia zerumbet (Pers.) B. L. Burtt & R. M.
Sm. and antimicrobial activity. Rev Bras Farmacogn 2009; 19(3): 697-701.
http://dx.doi.org/10.1590/S0102-695X2009000500008
http://dx.doi.org/10.1590/S0102-695X2009...
). C. sativum, belonging
to the Umbelliferae family, has antifungal activity (FURLETTI et al., 2011Furletti VF, Teixeira IP, Obando-Pereda G, Mardegan RC, Sartoratto
A, Figueira GM, et al. Action of Coriandrum sativum L. essential oil upon oral
Candida albicans biofilm formation. Evid-Based Complement Alternat Med 2011;
2011: 1-9. http://dx.doi.org/10.1155/2011/985832
http://dx.doi.org/10.1155/2011/985832...
), nematicidal activity against the phytonematode
Bursaphelenchus xylophilus (KIM
et al., 2008Kim J, Seo SM, Lee SG, Shin SC, Park IK. Nematicidal activity of
plant essential oils and components from coriander (Coriandrum sativum),
oriental sweetgum (Liquidambar orientalis), and valerian (Valeriana wallichii)
essential oils against pine wood nematode (Bursaphelenchus xylophilus). J Agric
Food Chem 2008; 56(16): 7316-7320.
http://dx.doi.org/10.1021/jf800780f
http://dx.doi.org/10.1021/jf800780f...
), and anthelmintic activity on gastrointestinal nematodes
(EGUALE et al., 2007Eguale T, Tilahun G, Debella A, Feleke A, Makonnen E. In vitro and
in vivo anthelmintic activity of crude extracts of Coriandrum sativum against
Haemonchus contortus. J Ethnopharmacol 2007; 110(3): 428-433.
http://dx.doi.org/10.1016/j.jep.2006.10.003
http://dx.doi.org/10.1016/j.jep.2006.10....
).
The objective of this study was to evaluate the effects of essential oils of L. camara, A. zerumbet, C. sativum and T. minuta on the egg hatching and larval development of H. contortus.
Materials and Methods
Plants and essential oils
L. camara, A. zerumbet and T. minuta were collected in Fortaleza, State of Ceará, Brazil, in campus of Universidade Estadual do Ceará in January of 2010. These plants were authenticated in the Herbarium Prisco Bezerra of the Universidade Federal do Ceará and voucher specimens were deposited under the numbers 46017, 49659, and 49676, respectively. The seeds of C. sativum were purchased commercially from the local market of Fortaleza.
To prepare essential oils, fresh aerial parts of 2.0 kg of L. camara, 2.7 kg of A. zerumbet, and 0.8 kg of T. minuta and 1.0 kg of seeds of C. sativum were separately subjected to hydrodistillation for 3 hours in a Clevenger-type apparatus. The calculation of yield was done and the oils were stored at 4 °C until use.
Egg hatch test
The egg hatch test (EHT) was performed based on the methodology
described by Coles et al. (1992)Coles GC, Bauer C, Borgsteede FHM, Geerts S, Klei TR, Taylor MA, et
al. World Association for the Advancement of Veterinary Parasitology
(W.A.A.V.P.) methods for the detection of anthelmintic resistance in nematodes
of veterinary importance. Vet Parasitol 1992; 44(1-2): 35-44.
http://dx.doi.org/10.1016/0304-4017(92)90141-U
http://dx.doi.org/10.1016/0304-4017(92)9...
. Sheep
experimentally infected with H. contortus were used as a source
of fresh eggs. H. contortus eggs were recovered according to
Hubert and Kerboeuf (1992)Hubert J, Kerboeuf D. A microlarval development assay for the
detection of anthelmintic resistance in sheep nematodes. Vet Rec 1992; 130(20):
442-446. http://dx.doi.org/10.1136/vr.130.20.442
http://dx.doi.org/10.1136/vr.130.20.442...
and Oliveira et al. (2009)Oliveira LMB, Bevilaqua CML, Costa CTC, Macedo ITF, Barros RS,
Rodrigues ACM, et al. Anthelmintic Activity of Cocos nucifera L. Against Sheep
Gastrointestinal Nematodes. Vet Parasitol 2009; 159(1): 55-59.
http://dx.doi.org/10.1016/j.vetpar.2008.10.018
http://dx.doi.org/10.1016/j.vetpar.2008....
. Briefly, 10 g of
feces, collected directly from the rectum, were mixed with distilled water and
filtered through 590, 149, 101 and 30 µm mesh sieves. To increase the
aqueous solubility, the oils were diluted in 3% Tween 80. An egg suspension (250
µL) containing approximately 100 fresh eggs was incubated with 250
µL essential oils at concentrations from 0.15 to 10 mg mL-1
for 48 h at 25 °C. Drops of Lugol were added. The eggs and first larval
stage (L1) were counted under a microscope. This test had two controls: a
negative control containing the diluent (3% Tween 80) and a positive control,
0.025 mg mL-1 of thiabendazole. Three repetitions with five
replicates for each essential oil concentration and for each control were
performed.
Larval development test
A larval development test (LDT) was performed according Camurça-Vasconcelos et al. (2007)Camurça-Vasconcelos ALF, Bevilaqua CML, Morais SM, Maciel MV, Costa
CTC, Macedo ITF, et al. Anthelmintic activity of Croton zehntneri and Lippia
sidoides essential oils. Vet Parasitol 2007; 148(3-4): 288-294.
http://dx.doi.org/10.1016/j.vetpar.2007.06.012
http://dx.doi.org/10.1016/j.vetpar.2007....
. For
this, H. contortus eggs were incubated for 24 h at 28 °C to
obtain the L1. Next, 1 mL of larval suspension containing approximately 250 L1
and 1 mL of essential oils at concentrations of 0.62 to 20 mg mL-1
were incubated with 2 g feces from a nematode-free sheep for 6 days at room
temperature. Then, the third-stage larvae (L3) were recovered according to Roberts and O'Sullivan (1950)Roberts FHS, O'Sullivan PJ. Methods for egg counts and larval
cultures for strongyles infecting the gastrointestinal tract of cattle. Aust J
Agric Res 1950; 1: 99-102. http://dx.doi.org/10.1071/AR9500099
http://dx.doi.org/10.1071/AR9500099...
and
counted under a microscope. This test had two controls, a negative with 3% Tween
80 and a positive with 0.008 mg mL-1 ivermectin. Three repetitions
with five replicates for each oil concentration and for each control were
conducted.
Chemical analysis
The chemical composition of the essential oils used in this study was determined by gas chromatography (GC) and mass spectrometry (MS). The oil was analyzed on a Hewlett-Packard 5971 instrument using the following experimental conditions: DB-1-coated fused silica capillary column (30m × 0.25 mm) - carrier gas - helio; injector temperature - 220 °C; detector temperature - 200 °C; column temperature program - 35-180 °C at 48 °C /min, then 180-250 °C at 10 °C/min. For MS, the electron impact was 70 eV.
Compounds were identified by their GC retention time, expressed by Kovat's index, which was calculated by the Van den Dool and Kratz equation using a hydrocarbon homologous series and by comparison of test compound mass spectra with those present in the National Institute for Standard Technology computer data bank (NIST; 62,235 compounds) and published spectra (ADAMS, 2001).
Statistical analysis
The effectiveness of each treatment on the EHT was determined based on the percentage of larvae, using the following formula: number of larvae/(number larvae + number eggs) × 100.
The inhibition percentage of the LDT was calculated based on the percentage reduction in the L3 recovered compared to the negative control group (number of L3 in the control group - number of L3 in the treated group)/number of L3 in control group × 100.
The results of the in vitro tests are expressed as the percentage of egg hatching or larval development inhibition. Data were analyzed using ANOVA and compared by the Tukey test (P<0.05) using the GraphPad Prism program 5.0. The effective concentrations that inhibited 50% (EC50) of egg hatching and larval development were determined by the probit method using SPSS 8.0 for Windows.
Results
The yields of the essential oil were 0.05% to L. camara, 0.14% to T. minuta, 0.24% from C. sativum and 0.38% to A. zerumbet.
The mean percentages of the effectiveness of the essential oils obtained in the EHT are presented in Table 1. L. camara was not effective against egg hatching at the concentrations tested. However, the oils of A. zerumbet, T. minuta and C. sativum present at concentrations ≥ 1.25 mg mL-1 egg hatch inhibition superior to negative control (P<0.05).
Table 2 shows the mean efficacy of essential oils by the LDT. The effective concentration of L. camara was 20 mg mL-1, did not differ from positive control (P>0.05). However, at a concentration of 10 mg mL-1, the A. zerumbet and C. sativum oils had similar efficacy to the positive control (P>0.05). Moreover, the effectiveness of 5 mg mL-1 T. minuta was not significantly different from the positive control (P>0.05).
The EC50 values for the EHT and LDT are shown in Table 3. The oils tested on the larvae and eggs of H. contortus showed a dose-dependent effect.
Effective concentration (mg.mL-1) to inhibit 50% (EC50) of egg hatch test (EHT) and larval development test (LDT) of Alpinia zerumbet, Coriandrum sativum, Tagetes minuta and Lantana camara essential oils on Haemonchus contortus.
The constituents of the essential oils are shown in Table 4. The major constituents of A. zerumbet oil were 1,8-cineole (24.69%), p-cimene (22.56%) and 4-terpineol (17.43%); L. camara contained caryophyllene oxide (50.26%); T. minuta contained piperitone (86.27%) and C. sativum contained beta linalool (73.21%).
Discussion
This study verified the presence of biologically active compounds that
had ovicidal effect on H. contortus in the essential oils of
A. zerumbet, C. sativum and T.
minuta. The oils inhibited egg hatching at a low concentration compared
to other plants that have been studied previously. Ocimum
gratissimum essential oil inhibited 100% of egg hatching at a
concentration of 50 mg mL-1 (PESSOA et
al., 2002Pessoa LM, Morais SM, Bevilaqua CM, Luciano JHS. Anthelmintic
activity of essential oil of Ocimun gratissimum Linn. and eugenol against
Haemonchus contortus. Vet Parasitol 2002; 109(1-2): 59-63.
http://dx.doi.org/10.1016/S0304-4017(02)00253-4
http://dx.doi.org/10.1016/S0304-4017(02)...
). The maximum effectiveness of the essential oil of
Eucalyptus globulus on eggs was 99.3% at a concentration of
21.75 mg mL-1 (MACEDO et al.,
2009Macedo ITF, Bevilaqua CML, Oliveira LMB, Camurça-Vasconcelos ALF,
Vieira LS, Oliveira FR, et al. Atividade ovicida e larvicida in vitro do óleo
essencial de Eucalyptus globulus sobre Haemonchus contortus. Rev Bras Parasitol
Vet 2009; 18(3): 62-66. http://dx.doi.org/10.4322/rbpv.01803011
http://dx.doi.org/10.4322/rbpv.01803011...
). In other study, the decoction of L. camara also did
not act on eggs and decoctions of A. zerumbet and T.
minuta exhibited inferior efficacy, inhibiting 97.5 and 96.8% of egg
hatching at a concentration 5 and 2.5 mg mL-1, respectively (MACEDO et al., 2012Macedo ITF, Bevilaqua CML, Oliveira LMB, Camurça-Vasconcelos ALF,
Morais SM, Machado LKA, et al. In vitro activity of Lantana camara, Alpinia
zerumbet, Mentha villosa and Tagetes minuta decoctions on Haemonchus contortus
eggs and larvae. Vet Parasitol 2012; 190(3-4): 504-509.
http://dx.doi.org/10.1016/j.vetpar.2012.07.001
http://dx.doi.org/10.1016/j.vetpar.2012....
). However, the crude
aqueous and hydro-alcoholic extracts of the seeds of C. sativum
completely inhibited the hatching of egg at a concentration of less than 0.5 mg
mL-1. Phytochemical screening of these extracts revealed the presence
of secondary metabolites such as alkaloids and flavonoids, which are considered to
be the chemical components that are responsible for the wide therapeutic activities
of several medicinal plants (EGUALE et al.,
2007Eguale T, Tilahun G, Debella A, Feleke A, Makonnen E. In vitro and
in vivo anthelmintic activity of crude extracts of Coriandrum sativum against
Haemonchus contortus. J Ethnopharmacol 2007; 110(3): 428-433.
http://dx.doi.org/10.1016/j.jep.2006.10.003
http://dx.doi.org/10.1016/j.jep.2006.10....
). Variation in activity between the extract types and essential oil
might be due to differences in the proportion of the active components responsible
for the tested anthelmintic activity.
The most potent oil tested in LDT was T. minuta, which
exhibited superior efficacy to other oils tested previously with an EC50 of 1.67 mg
mL-1. The essential oil of Eucalyptus citriodora had
an EC50 of 2.71 mg mL-1 (MACEDO et al.,
2011Macedo ITF, Bevilaqua CML, Oliveira LMB, Camurça-Vasconcelos ALF,
Vieira LS, Amóra SSA. Avaliação do óleo essencial de Eucalyptus citriodora sobre
nematóides gastrintestinais de caprinos. Rev Bras Parasitol Vet 2011; 20(3):
223-227. http://dx.doi.org/10.1590/S1984-29612011000300009
http://dx.doi.org/10.1590/S1984-29612011...
). The EC50 of Lippia sidoides essential oil was
2.97 mg mL-1 (CAMURçA-VASCONCELOS et al.,
2007Camurça-Vasconcelos ALF, Bevilaqua CML, Morais SM, Maciel MV, Costa
CTC, Macedo ITF, et al. Anthelmintic activity of Croton zehntneri and Lippia
sidoides essential oils. Vet Parasitol 2007; 148(3-4): 288-294.
http://dx.doi.org/10.1016/j.vetpar.2007.06.012
http://dx.doi.org/10.1016/j.vetpar.2007....
). However, T. minuta essential oil was similar to
Eucalyptus staigeriana essential oil, which had an EC50 of 1.70
mg mL-1 (MACEDO et al., 2010Macedo ITF, Bevilaqua CML, Oliveira LMB, Camurça-Vasconcelos ALF,
Vieira LS, Oliveira FR, et al. Anthelmintic effect of Eucalyptus staigeriana
essential oil against goat gastrointestinal nematodes. Vet Parasitol 2010;
173(1-2): 93-98. http://dx.doi.org/10.1016/j.vetpar.2010.06.004
http://dx.doi.org/10.1016/j.vetpar.2010....
).
Transcuticular diffusion is a common method of entry into in helminth
parasites for non-nutrient and non-electrolyte substances (EGUALE et al., 2007Eguale T, Tilahun G, Debella A, Feleke A, Makonnen E. In vitro and
in vivo anthelmintic activity of crude extracts of Coriandrum sativum against
Haemonchus contortus. J Ethnopharmacol 2007; 110(3): 428-433.
http://dx.doi.org/10.1016/j.jep.2006.10.003
http://dx.doi.org/10.1016/j.jep.2006.10....
). It has also been shown that this route,
rather than oral ingestion, predominates for the uptake of major broad-spectrum
anthelmintics. It is easier for lipophilic anthelmintics to cross the external
surface of helminths than it is for hydrophilic compounds (GEARY et al., 1999Geary TG, Sangster NC, Thompson DP. Frontiers in anthelmintic
pharmacology. Vet Parasitol 1999; 84(3-4): 275-295.
http://dx.doi.org/10.1016/S0304-4017(99)00042-4
http://dx.doi.org/10.1016/S0304-4017(99)...
). The low density of plant oils and their
rapid diffusion across cell membranes can enhance the targeting of the active
components of essential oils into endoparasites (ANTHONY et al., 2005Anthony JP, Fyfe L, Smith H. Plant active components - a resource
for antiparasitic agents? Trends Parasitol 2005; 21(10): 462-468.
http://dx.doi.org/10.1016/j.pt.2005.08.004
http://dx.doi.org/10.1016/j.pt.2005.08.0...
).
Essential oils are composed of a mixture of chemical substances whose interaction can result in compounds that can interfere with nematode metabolism, inhibiting or disorganizing vital functions from the initial stages of development onward, and can furthermore interfere with drive mechanisms due to possible destructuring of the nervous system (OKA et al., 2000Oka Y, Nacar S, Putievsky E, Ravid U, Zohara Y, Spiegel Y. Nematicidal activity of essential oils and their components against the root-knot nematode. Phytopathology 2000; 90(7): 710- 715.). Essential oils can interact with the cytoplasmic membrane and may disrupt the structure of polysaccharides, lipids and phospholipids, promoting membrane depolarization and the consequent alteration of its permeability (BAKKALI et al., 2008), as reported for fungi exposed to essential oils (HAMMER et al., 2004Hammer KA, Carson CF, Riley TV. Antifungal effects of Melaleuca alternifolia (tea tree) oil and its components on Candida albicans, Candida glabrata and Saccharomyces cerevisiae. J Antimicrob Chemother 2004; 53(6): 1081-1085.).
Chemical analysis identified components with high concentrations in
essential oils that may be responsible for their anthelmintic activity. The main
component of A. zerumbet was 1,8-cineol, which has previously been
shown to have antibacterial activity (CHA et al.,
2007Cha JD, Jung EK, Kil BS, Lee KY. Chemical composition and
antibacterial activity of essential oil from Artemisia feddei. J Microbiol
Biotechnol 2007; 17(12): 2061-2065.), antifungal activity (TERZI et al.,
2007Terzi V, Morcia C, Faccioli P, Vale G, Tacconi G, Malnati M. In
vitro antifungal activity of the tea tree (Melaleuca alternifolia) essential oil
and its major components against plant pathogens. Lett Appl Microbiol 2007;
44(6): 613-618.
http://dx.doi.org/10.1111/j.1472-765X.2007.02128.x
http://dx.doi.org/10.1111/j.1472-765X.20...
) and activity against the nematode Anisakis simplex
(NAVARRO et al., 2008Navarro MC, Noguera MA, Romero MC, Montilla MP, Selgas JMG, Valero
A. Anisakis simplex s.l.: Larvicidal activity of various monoterpenic
derivatives of natural origin against L3 larvae in vitro and in vivo. Exp
Parasitol 2008; 120(4): 295-299.
http://dx.doi.org/10.1016/j.exppara.2008.07.014
http://dx.doi.org/10.1016/j.exppara.2008...
). The main
component of L. camara was caryophyllene oxide, an antifungal agent
(YANG et al., 1999Yang D, Michel L, Chaumont JP, Millet-Clerc J. Use of caryophyllene
oxide as an antifungal agent in an in vitro experimental model of onychomycosis.
Mycopathologia 1999; 148(2): 79-82.) that has analgesic
and anti-inflammatory activity (CHAVAN et al.,
2010Chavan MJ, Wakte PS, Shinde DB. Analgesic and anti-inflammatory
activity of Caryophyllene oxide from Annona squamosa L. bark. Phytomedicine
2010; 17(2): 149-151.
http://dx.doi.org/10.1016/j.phymed.2009.05.016
http://dx.doi.org/10.1016/j.phymed.2009....
). The main component of C. sativum was linalool,
which has antibacterial activity (FISHER et al., 2007) and insecticidal activity
against Tribolium castaneum (STAMOPOULOS et al., 2007Stamopoulos DC, Damos P, Karagianidou G. Bioactivity of five
monoterpenoid vapours to Tribolium confusum (du Val) (Coleoptera :
Tenebrionidae). J Stored Prod Res 2007; 43(4): 571-577.
http://dx.doi.org/10.1016/j.jspr.2007.03.007
http://dx.doi.org/10.1016/j.jspr.2007.03...
), Oryzaephilus surinamensis,
Musca domestica and Blattella germanica (LEE et al., 2003Lee S, Peterson CJ, Coats JR. Fumigation toxicity of monoterpenoids
to several stored product insects. J Stored Prod Res 2003; 39: 77-85.
http://dx.doi.org/10.1016/S0022-474X(02)00020-6
http://dx.doi.org/10.1016/S0022-474X(02)...
), and nematicidal activity
against juveniles of the root-knot nematode, Meloidogyne incognita
(LEELA et al., 1992Leela NK, Khan RM, Reddy PP, Nidiry ESJ. Nematicidal activity of
essential oil of Pelargonium graveolens against the root-knot nematode
Meloidogyne incognita. Nematol Medit 1992; 20(1): 57-58.). The major
constituent of T. minuta, piperitone was an antioxidant (DAR et al., 2011Dar MY, Shah WA, Rather MA, Qurishi Y, Hamid A, Qurishi MA. Chemical
composition, in vitro cytotoxic and antioxidant activities of the essential oil
and major constituents of Cymbopogon jawarancusa (Kashmir). Food Chem 2011;
129(4): 1606-1611.
http://dx.doi.org/10.1016/j.foodchem.2011.06.016
http://dx.doi.org/10.1016/j.foodchem.201...
), which has insecticidal
activity against larvae of Spodoptera littoralis and fungicidal
activity (ABDELGALEIL et al., 2008Abdelgaleil SAM, Abbassy MA, Belal ASH, Rasoul MAAA. Bioactivity of
two major constituents isolated from the essential oil of Artemisia judaica L.
Bioresour Technol 2008; 99(13): 5947-5950.
http://dx.doi.org/10.1016/j.biortech.2007.10.043
http://dx.doi.org/10.1016/j.biortech.200...
).
The plants evaluated in the current study had already been reported as anthelmintic agents. Therefore, the current finding is the first step to justify their use in folk medicine. Further investigation of isolated fractions at different dose levels should be pursued. In addition, tests with animals can be performed with A. zerumbet, C. sativum and T. minuta essential oils to evaluate the toxicity and to confirm their anthelmintic activity in target species.
This work received financial support from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico. Dr. Bevilaqua has a grant from CNPq.
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Publication Dates
-
Publication in this collection
Oct-Dec 2013
History
-
Received
28 Feb 2013 -
Accepted
24 June 2013