In vitro and in vivo activity of the essential oil and nanoemulsion of Cymbopogon flexuosus against Trichomonas gallinae

Document Type : Original Research Article


1 Department of Microbiology and Parasitology- Federal University of Pelotas, Pelotas, RS, Brazil

2 Laboratory of Nanotechnology - Franciscan University, Santa Maria, RS, Brazil


Objective: This study was done to evaluate the in vitro and in vivo effects of the essential oil (OE-CL) and nanoemulsion (N-CL) of Cymbopogon flexuosus against Trichomonas gallinae.
Materials and Methods: In vitro assays were done with 106 parasites and OE-CL and N-CL in the concentrations: 110, 220, 330, 440, 550, 660, 770 and 880 µg/ml and four controls: CN (culture medium and trophozoites), MTZ (trophozoites plus 800 µg/ml of metronidazole), TW (trophozoites plus vehicles used for solubilization of derivatives (0.01% Tween) and NB (blank nanoemulsion 880 µg/ml). The in vivo assay was done in 35 quails (Coturnix coturnix) infected experimentally 4x104 mg/kg,  were divided in   seven groups (n=5): A (control–healthy), B (control infected), C (control TW 0.01%), D (NB 0.88 mg/kg), E (drug MTZ 25 mg/kg, F (OE-CL at 0.55 mg/kg ) and G (N-CL at 0.44 mg/kg), during 7 consecutive days.
Results: The in vitro test showed that the OE-CL (550 μg/ml) and N-CL (440 μg/ml) concentrations reduced the trophozoites viability in 100%. In the in vivo test, the treatment with OE-CL was efficient on the 4th treatment day and the N-CL after the 3rd day, and the MTZ in the therapeutic concentration was efficient on the 7th day.
Conclusion: It can be observed in this study that the lemon grass has natural potential antitrichomonal activity against T. gallinae in vitro and in vivo.


Adukwu EC, Allen SC, Phillips CA. 2012. The
anti-biofilm activity of lemongrass
(Cymbopogon flexuosus) and grapefruit
(Citrus paradisi) essential oils against five
strains of Staphylococcus aureus. J Appl
Microbiol, 113:1217-27.
Ahmad A, Viljoen A. 2015. The in vitro
antimicrobial activity of Cymbopogon
essential oil (lemon grass) and its interaction
with silver ions. Phytomedicine, 22:657-
Bahamonde-Norambuena D, Molina-Pereira A,
Cantin M, Muñoz M, Zepeda K, Vilos C.
2015. Polymeric nanoparticles in
dermocosmetic. Int J Morphol, 33:1563-
Bondurant RH, Honigberg BM. 1994.
Trichomonads of veterinary importance. In:
Academic Press (Eds.), Parasitic Protozoa,
p. 111–206, New York.
Bunbury N, Jones CG, Greenwood AG, Bell
DJ. 2007. Trichomonas gallinae in
Mauritian Columbids: Implications for an
endangered endemic. J Wild Dis, 43:399–
Cepicka I, Kutišová K, Tachezy J, Kulda J,
Flegr J. 2005 Cryptic species within the
Tetratrichomonas gallinarum species
complex revealed by molecular
polymorphism. Vet Parasitol, 128:11–21.
Cudmore SL, Delgaty KL, HaywardMcClelland SF, Petrin DP, Garber GE.
2004. Treatment of infections caused by
metronidazole-resistant Trichomonas
vaginalis. Clin Microbiol Rev, 17:783-93.
Diamond, LS. 1957. The establishment of
various trichomonads of animals and man in
axenic cultures. J Parasitol, 43:488-490.
Dingsdag SA, Hunter N. 2018. Metronidazole:
an update on metabolism, structure–
cytotoxicity and resistance mechanisms. J
Antimicrob Chemother, 73:265-79.
Dunne RL, Dunn LA, Upcroft P, O’Donoghue
PJ, Upcroft JA. 2003. Drug resistance in the
sexually transmitted protozoan
Trichomonas vaginalis. Cell Res, 13:239-
Fang L, Du D, Ding GZ, Si YK, Yu SS, Liu Y,
Wang WJ, Ma SG, Xu S, Qu J, Wang JM,
Liu YX. 2010. Neolignans and glycosides
from the stem bark of Illicium difengpi. J
Nat Product, 73: 818-824.
Forrester DJ, Foster GW. 2009. Trichomonosis.
In: Atkinson, C.T., Thomas, N.J.; Hunter,
D.B. (Eds.), Parasitic Diseases of Wild
Birds, pp.120-153, Oxford, UK: WileyBlackwell.
Franssen FFJ, Lumeij JT. 1992. In vitro
nitroimidazole resistance of Trichomonas
gallinae and successful therapy with an
increased dosage of ronidazole in racing
pigeons (Columba livia domestica). J Vet
Pharmacol Therapeut, 15:409–415.
Ganjewala D1, Luthra R. 2010. Essential oil
biosynthesis and regulation in the genus
Cymbopogon. Nat Prod Commun, 5:163-72.
Gaspar da Silva D, Barton E, Bunbury N,
Lunness P, Bell DJ, Tyler KM. 2007.
Molecular identity and heterogeneity of
trichomonad parasites in a closed avian
population. Infect Genet Evolut, 7:433-440.
Gerhold RW, Tate CM, Gibbs SE, Mead DG,
Allison AB, FISCHER JR. 2007. Necropsy
findings and arbovirus surveillance in
mourning dovesfrom southeastern United
States. J Wild Dis, 43:129-135.
Gündel SS, de Souza MB, Quatrin PM, Klein
B, Wagner R, Gündel A, Vaucher R de A,
Santos RCV, Ourique AF. 2018.
Nanoemulsions containing Cymbopogon
flexuosus essential oil: Development,
characterization, stability study and
evaluation of antimicrobial and antibiofilm
activities. Microb Pathog, 118:268-276.
Hosseini SF, Zandi M, Rezaei M,
Farahmandghavi F. 2013. Two-step method
for encapsulation of oregano essential oil in
chitosan nanoparticles: preparation,
characterization and in vitro release study.
Carbohydr Polym, 95:50-56.
Hussain AI, Anwar F, Shahid M, Ashraf M,
Przybylski R, 2010. Chemical composition,
antioxidant and antimicrobial activities of
essential oil of spearmint (Mentha spicata
L.) from Pakistan. J Essent Oil Res, 22:78-
Hussein R, El-Halabi M, Ghaith O, Jurdi N,
Baccega et al.
AJP, Vol. 11, No. 1, Jan-Feb 2021 42
Azar C, Mansour N, Sharara AI. 2011.
Severe hepatotoxicity associated with the
combination of spiramycin plus
metronidazole. Arab J Gastroenterol, 12:44–
Kumar S, Stecher G, Li M, Knyaz C, Tamura
K. 2018. MEGA X: Molecular Evolutionary
Genetics Analysis across computing
platforms. Mol Biol Evolut, 35:1547-1549.
Lumeij JT, Zwijnenberg RJG. 1990. Failure of
nitro-imidazole drugs to control
trichomoniasis in the racing pigeon
(Columba livia domestica). Avian Pathol,
Mehlhorn H, Walldorf V, Klimpel S, Schmahl
G, Al-Quraishy A, Walldorf U, Mehlhorn B,
Bätza HJ. 2009. Entomological survey on
vectors of Bluetongue virus in North RhineWestphalia (Germany during 2007–2008).
Parasitol Res, 102:321-9.
Munoz E, Castella J, Gutierrez J. 1998. In vivo
and sensitivity of Trichomonas gallinae to
some nitroimidazole drugs. Vet Parasitol,
31: 239-46.
Nair J, Christensen D, Yu P, Beattie AD,
McAllister T, Damiran D, Preston N, Fuhr
L, McKinnon JJ. 2016. A nutritional
evaluation of common barley varieties
grown for silage by beef and dairy producers
in western Canada. Can J Anim Sci, 96:598-
Oliveira MMM, Brugnera DF, Nascimento JA
& Piccoli RH. 2012. Control of planktonic
and sessile bacterial cells by essential oils.
Food and Bioproducts Processing, 90:809-
Rein MF. Trichomonas vaginalis. 1995. In:
Mandell, Douglas, Bennet. Principles and
practice of infectious diseases (Eds.)
Churchill Livingstone, 2493-7, New York:
Rouffaer L, Adriaensen C, de Boeck C,
Claerebout E, Martel A. 2014. Racing
pigeons: a reservoir for nitro-imidazoleresistant Trichomonas gallinae. J Parasitol,
Salem W, Leitner DR, Zingl FG, Schratter G,
Prassl R, Goessler W, Reidl J, Schild S.
2015. Antibacterial activity of silver and
zinc nanoparticles against Vibrio cholerae
and enterotoxic Escherichia coli. Int J Med
Microbiol, 305:85–95.
Sansano-Maestre J, Garijo-Toledo MM,
Gómez-Muñoz MT. 2009. Prevalence and
genotyping of Trichomonas gallinae in
pigeons and birds of prey. Avian Pathol, 38:
Santoro GF, Cardoso MG, Guimarães LG,
Salgado AP, Mena-Barreto RF, Soares MJ
2007b. Effect of oregano (Origanum
vulgare L.) and thyme (Thymus vulgaris L.)
essential oils on Trypanosoma cruzi
(Protozoa: Kinetoplastida) growth and
ultrastructure. Parasitol Res 100: 783-790.
Seddiek A, El-Shorbagy MM, Khater HF, Ali
AM. 2014. The antitrichomonal efficacy of
garlic and metronidazole against
Trichomonas gallinae infecting domestic
pigeons. Parasitol Res, 113:1319–1329.
Seddiek SA, Ali MM, Khater HF, El-Shorbagy
MM. 2011 Anthelmintic activity of the
white wormwood, Artemisia herba-alba
against Heterakis gallinarum infecting
turkey poults. J Med Plant Res, 5:3946–
Sena-Lopes Â, das Neves RN, Bezerra FSB, de
Oliveira Silva MT, Nobre PC, Perin G,
Alves D, Savegnago L, Begnini KR, Seixas
FK, Collares T, Borsuk S. 2017.
Antiparasitic activity of 1,3-dioxolanes
containing tellurium in Trichomonas
vaginalis. Biomed Pharmacot, 89: 284-287.
Shukla R, Kumar A, Singh P, Dubey NK. 2009.
Efficacy 1 of Lippia alba (Mill) N. E. Brown
essential oil and its monoterpenes aldehyde
constituents aganst fungi isolated from some
edible legume seeds and aflatoxin B1
production. Int J Food Microbiol, 135:165-
Silva CB, Guterres SS, Weisheimer V,
Schapoval E.E. 2008. Antifungal activity of
the lemongrass oil and citral against
Candida spp. Braz J Inf Dis, 12:1763- 1766.
Solans C, Izquierdo P, Nolla J, Azemar N,
Garcia-Celma MJ. 2005. Nanoemulsions.
Curr Opin Colloid Int, 10:102-110.
Sonneville-Aubrun O, Simonnet JT, L'alloret F.
2004. Nanoemulsions a new vehicle for
skincare products. Adv Col Interf Sci, 108-
Sun H, Liu K, Liu W, Wang W, Guo C, Tang
B, Gu J, Zhang J, Li H, Mao X, Zou Q, Zeng
H. 2012. Development and characterization
of a novel nanoemulsion drugdelivery
system for potential application in oral
delivery of protein drugs. Int J Nanomed,
Tabari MA, Youssefi MR, Moghadamnia AA.
2017. Antitrichomonal activity of Peganum
harmala alkaloid extract against
trichomoniasis in Pigeon (Columba livia
In vitro and in vivo activity of the Cymbopogon flexuosus on Trichomonas gallinae
AJP, Vol. 11, No. 1, Jan-Feb 2021 43
domestica), Br Poult Sci, 58: 236-241.
Tao N, OuYang Q, Jia L. 2014. Citral
inhibitsmycelial growth of Penicillium
italicum by a membrane damage
mechanism. Food Control, 41:116–121.
Tracy JW, Webster LT. 1996.Drugs used in the
chemoterapy of protozoal infections. In:
Goodman and Gilman's (Eds.), The
pharmacological basis of therapeutics.:
McGraw-Hill Companies, p. 995-8, New
Trados T, Izquierdo P, Esquena J, Solans C.
2004. Formation and stability of nano
emulsions. Adv coll Interf Sci, 108:303-318.
Upcroft P, Upcroft JA. 2001 Drug targets and
mechanisms of resistance in the anaerobic
protozoa. Clin Microbiol Ver, 14:150-64.
Wen-Chien L, Huang DW, Wang CR, Yeh CH,
Tsai JC, Huang YT, Li PH. 2018.
Preparation, characterization, and
antimicrobial activity of nanoemulsions
incorparating citral essential oil. J Food
Grug anal, 26:82-89.
Xavier MN, Alves JM, Carneiro NS, Souchie
EL, Silva EAJ, Martins CHG, Ambrosio
MALV, Egea MB, Alves CCF, Miranda
MLD. 2016. Composição química do óleo
essencial de Cardiopetalum calophyllum
Schltdl. (Annonaceae) e suas atividades
antioxidante, antibacteriana e antifúngica.
Rev Virt Quím, 8:1433.
Youssefi MR, Tabari MA, Moghadamnia AA.
2017. In vitro and in vivo activity of
Artemisia sieberi against Trichomonas
gallinae. Iran J Veterin Res, 18:25.
Zhou H, Tao N, Jia L. 2014. Antifungal activity
of citral, octanal and 𝛼-terpineol against
Geotrichumcitri-aurantii,” Food Control,
37: 277–283.
Zimre-Grabensteiner E, Arshad N, Amin A,
Hess M. 2011. Genetically different clonal
isolates of Trichomonas gallinae, obtained
from the same bird, can vary in their drug
susceptibility, an in vitro evidence. Parasitol
Int, 60:213-5.