Effect of Echinophora platyloba , Stachys lavandulifolia, and Eucalyptus camaldulensis plants on Trichomonas vaginalis growth in vitro
Hossein Ali Youse1, Afsaneh Kazemian2, Manijeh Sereshti2, Elham Rahmanikhoh2, Elham Ahmadinia2, Mahmood Rafaian2, Reza Maghsoodi3, Hossein Y Darani1
1 Department of Parasitology and Mycology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
2 Department of Microbiology, Plant Research Center, Shahrekord, Iran
3 Faculty of Medicine, Shahrekord University of Medical Sciences, Shahrekord, Iran
|Date of Submission||22-Feb-2012|
|Date of Acceptance||03-Jun-2012|
|Date of Web Publication||31-Oct-2012|
Hossein Y Darani
Department of Parasitology and Mycology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan
Source of Support: None, Conflict of Interest: None
Background: Trichomonas vaginalis is a protozoan parasite which causes vaginitis in women worldwide. Metronidazole with vast side effects is drug of choice for this infection. In search for an alternative drug, in this study the effect of three plants on Trichomonas vaginalis has been investigated in vitro.
Materials and Methods: Alcoholic and watery extracts of Echinophora platyloba, Stachys lavandulifolia, and Eucalyptus camaldulensis were prepared. In TYIS culture medium containing alive Trichomonas vaginalis different concentrations of extracts of three plants were added. Following, 24, 48, and 72 h incubation the number of parasite in each test tube was counted.
Results: Eucalyptus camaldulensis showed a strong effect on Trichomonas vaginalis growth. However, no significant effect was observed with Echinophora platyloba or Stachys lavandulifolia extracts.
Conclusion: Eucalyptus camaldulensis can be considered as an alternative drug for treatment of infective vaginitis which is caused by bacteria, fungi and parasites.
Keywords: Echinophora platyloba , Eucalyptus camaldulensis, plants, Stachys lavandulifolia, Trichomonas vaginalis
|How to cite this article:|
Youse HA, Kazemian A, Sereshti M, Rahmanikhoh E, Ahmadinia E, Rafaian M, Maghsoodi R, Darani HY. Effect of Echinophora platyloba , Stachys lavandulifolia, and Eucalyptus camaldulensis plants on Trichomonas vaginalis growth in vitro. Adv Biomed Res 2012;1:79
|How to cite this URL:|
Youse HA, Kazemian A, Sereshti M, Rahmanikhoh E, Ahmadinia E, Rafaian M, Maghsoodi R, Darani HY. Effect of Echinophora platyloba , Stachys lavandulifolia, and Eucalyptus camaldulensis plants on Trichomonas vaginalis growth in vitro. Adv Biomed Res [serial online] 2012 [cited 2019 Jul 20];1:79. Available from: http://www.advbiores.net/text.asp?2012/1/1/79/102987
| Introduction|| |
Trichomonas vaginalis , a protozoan parasite, is a common sexually transmitted infection in women and men. , It is usually transmitted by sexual relationship, by sharing contaminated underwear clothes or by nonhygienic vaginal examination.  In women, it causes vaginitis and cystitis and in men it causes urethritis and prostatitis.  Trichomonas vaginalis is considered as a common cause of vaginitis and as a causative factor for preterm birth and low birth weight.  It has also been associated with increased human immunodeficiency virus transmission. 
Metronidazole with vast side effects is now considered as a drug of choice for this infection with a cure rate of approximately 95%.  Clinical resistance to this drug has been reported since 1962.  Therefore, it would be very important to search for an alternative drug.
Echinophora plants have four species in Iran and distributed in the west and western north of Iran.  The genus Stachys is widespread throughout the world. In Iran, many species of this genus are present, from which, 13 are endemic. Stachys lavanduifolia has been used as an anxiolytic and sedative in Iranian folk medicine.  Analysis of the crude extracts of the Eucalyptus spp. revealed the presence of saponin, saponin glycosides, steroid, cardiac glycoside, tannins, volatile oils, phenols and balsam. 
Antibacterial, antifungal or antiparasitic effects of Echinophora platyloba,,,,,, Stachys lavandulifolia, , and Eucalyptus camaldulensis, plants have been reported. In some local area of Iran these plants may be used for vaginal infection treatment. In this work, the effect of Echinophora platyloba, Stachys lavandulifolia, and Eucalyptus camaldulensis with antiparasitic activity  on Trichomonas vaginalis has been investigated in vitro.
| Materials and Methods|| |
Echinophora platyloba , Stachys lavandulifolia were collected from Chahar Mahal va Bakhtiari mountains and Eucalyptus camaldulensis was collected form Khostan trees. All plants were collected in spring of 2010 and confirmed by experts in medical plant research center of Shahrekord university o f medical sciences. Leaves of those three plants were washed with water, dried, and then ground finely in a spice small electric mill. The resulting powders were mixed with ethanol or water to make extractions.
In order to prepare alcoholic extracts, 40 g of each dried powder plant was added to 1 L of 96% ethanol and left for 12 days. The mixture clarified by filtration through a sterile filter paper. The product was placed in the vacuum rotary evaporation device to remove solvent alcohol and it was dried in the 40°C oven.
In order to prepare watery extracts, 40 g of each dried powder plant soaked in 1 L distilled water and left for 48 h. The mixture clarified by filtration through a sterile filter paper. Vacuum rotary evaporation device was used to remove the solvent water. Stock solutions of dried extracts were reconstituted in 50% DMSO.
Trichomonas vaginalis parasite was isolated from vaginal discharge of women referred to Hajar hospital clinic in Shahrekord city, Iran and transferred to TYIS culture medium. A pooled of five parasite isolates was used for experiments. In test tubes containing 10 ml of TYIS culture medium different concentrations of dried extracts of three plants were added. In one tube metronidazole (5 μg/ml) was added as positive control and one tube left intact as negative control. Then 50 μl of medium containing about 100 live Trichomonas vaginalis were added to each tube. All tubes incubated at 37°C and the number of parasites in each tube was counted following, 24, 48, and 72 h incubation. For parasite counting, each tube was first shacked and 10 μl of it was observed on a microscope slide. Trichomonas vaginalis parasites with active flagella were considered alive. Each experiment was performed in triplicate.
| Results|| |
In tubes contained metronidazole (5 μg /ml), 60 or 90 μg Eucliptus, no parasite was observed after 72 h, while in control test tubes parasite had a normal growth. Details of parasite counts in all test tubes have been presented in [Table 1]. No significant effect on Trichomonas vaginalis growth was observed when alcoholic or watery extract of aerial parts of Stachys lavandulifolia were used. Details of parasite counts of this experiment presented in [Table 2]. Also no significant effect was observed when Echinophora platyloba was used.
|Table 1: Results of Trichomonas vaginalis counts in test tubes treated with Eucaliptus extract in comparison with appropriate controls|
Click here to view
|Table 2: Number of Trichomonas vaginalis in 20 µl of TYIS-33 culture medium 72 h after treatment with different concentrations of Stachys lavandulifolia extracts or metronidazole|
Click here to view
| Discussion|| |
Results of this investigation revealed that Eucalyptus camaldulensis but not Echinophora platyloba or Stachys lavandulifolia had a strong effects on Trichomonas vaginalis growth in vitro.
Antimicrobial, antifungal, and antiparasitic effects of Eucaliptus spp. have been shown in different investigations. ,,, Adeniyi demonstrated that Eucaliptus camalddlensis had a strong effect on Helicobcter pylori in vitro.  Drug of choice for these bacteria is metronidazole, same drug for treatment of Trichomonas vaginalis. In another study Safaei et al. showed that essential oil of Eucaliptus spp. had a strong antimicrobial activities.  Nathan et al. also showed that essential oil of Eucaliptus spp. possess anti trypanosomal activity in vitro and this effects was dose dependent.  Ramazani et al. demonstrated antifungal activity of Eucaliptus spp.  Satorelli et al. investigated antimicrobial and antifungal activities essential oil of Eucaliptus Spp. They demonstrated that this plant presented the highest growth inhibition against Staphylococcus aureus, Esherichia coli, and Candida albicans.  Mahdi et al. studied effects of two plants, Myrtus communcs and Eucaliptus comaldensis, on Trichomonas vaginalis growth in vitro. They showed that Eucaliptus extract caused death of parasite at pH 5.3.  Results of this investigation are in agreement with our findings.
| Conclusion|| |
According to results of present investigation Eucalyptus camaldulensis plant could be considered as an alternative drug for Trichomonas vaginalis treatment. So it will be very worthwhile to recognize the efficient components of this plant with anti Trichomonas activities in further investigation. Moreover, antifungal and antiparasitic activities have been reported for Eucaliptus spp. ,,,,, Therefore this plant has a potential to be considered as a unique drug for treatment of infective vaginitis which is caused by bacteria, fungi or parasites. Further investigations are recommended to test this hypothesis.
| Acknowledgments|| |
This work was supported by a grant from the Shahrekord University of Medical Sciences, Iran.
| References|| |
|1.||Krieger JN. Consider diagnosis and treatment of trichomoniasis in men. Sex Transm Dis 2000;27:241-2. |
|2.||Petrin D, Delgaty K, Bhatt R, Garber G. Clinical and microbiological aspects of Trichomonas vaginalis. Clin Microbiol Rev 1998;11:300-17. |
|3.||King A, Nicol C. Venereal diseases. 7 & 8 Henrietta Street, London WC2E 8QE: Bailliére Tindall; 1975. |
|4.||Kuberski T. Trichomonas vaginalis associated with nongonococcal urethritis and prostatitis. Sex Transm Dis;1980;7:135-136. |
|5.||Cotch M, Pastorek JG, Nugent RP, Hillier SL, Gibbs RS, Martin DH, et al. Trichomonas vaginalis associated with low birth weight and preterm delivery. Sex Transm Dis 1997;24:353-60. |
|6.||Sorvillo F, Kovacs A, Kerndt P, Stek A, Muderspach L, Sanchez-Keeland L. Risk factors for trichomoniasis among women with human immunodeficiency virus (HIV) infection at a public clinic in Los Angeles County, California: Implications for HIV prevention. Am J Trop Med Hyg 1998;58:495-500. |
|7.||Durel P, Roiron V, Siboulet A, Borel LJ. Systemic treatment of trichomoniasis with a derivative of nitro-imidazole, 8823 RP. Br J Vener Dis 1960;36:21-6. |
|8.||Meingassner JG, Thurner J. Strain of Trichomonas vaginalis resistant to metronidazole and other 5-nitroimidazoles. Antimicrob Agents Chemother 1979;15:254-7. |
|9.||Entezari M, Hashemi M, Ashki M, Ebrahimian S, Bayat M, Azizi Saraji A. Studying the effect echinophora platyloba extract on bactira (staphilococus aureus and pseudomonas aeroginosa) and Fungi (Candidia albicans, Aspergilus flavus and Aspergilus niger) in vitro. World J Med Sci 2009;4:89-92. |
|10.||Rabbani M, Sajjadi S, Zarei H. Anxiolytic effects of Stachys lavandulifolia on the elevated plus-maze model of anxiety in mice. J Ethnopharmacol 2003;89:271-6. |
|11.||Falahati M, Tabrizib NO, Jahaniani F. Anti dermatophyte activities of Eucalyptus camaldulensis in comparison with Griseofulvin. Iranian Journal of Pharmacology & Therapeutics (IJPT) 2005;80-3. |
|12.||Adeniyi CB, Lawal TO, Mahady GB. In vitro susceptibility of Helicobacter pylori to extracts of Eucalyptus camaldulensis and Eucalyptus torelliana. Pharm Biol 2009;47:99-102. |
|13.||Javad SG, Atefeh A. Antimicrobial and antifungal properties of the essential oil and methanol extracts of Eucalyptus largiflorens and Eucalyptus intertexta. Pharmacogn Mag 2010;6:172-5. |
|14.||Mahdi N, Gany Z, Sharief M. Alternative drugs against Trichomonas vaginalis. East Mediterr Health J 2006;12:679-84. |
|15.||Nathan H, Abel S. Evaluation of in vitro activity of essential oils against trypanosoma brucei brucei and Trypanosoma evansi. J Parasitol Res 2010. |
|16.||Ramezani H, Singh H, Batish D, Kohli R. Antifungal activity of the volatile oil of Eucalyptus citriodora. Fitoterapia 2002;73:261-2. |
|17.||Sartorelli P, Marquioreto AD, Amaral Baroli A, Lima ME, Moreno PR. Chemical composition and antimicrobial activity of the essential oils from two species of Eucalyptus. Phytother Res 2007;21:231-3. |
|18.||Ghasemi Pirbalouti A, Malekpoor F, Enteshari S, Yousefi M, Momtaz H, Hamedi B. Antibacterial activity of some folklore medicinal plants used by Bakhtiari tribal in Southwest Iran. International Journal of Biology 2010; 55-64. |
|19.||Saeedi M, Morteza-Semnani K, Mahdavi M, Rahimi F. Antimicrobial studies on extracts of four species of Stachys. Indian J Pharm Sci 2008;70:403-6. |
|20.||Avijgan M, Saadat M, Nilforoshzadeh M, Hafizi M. Antifungal effect of echinophora platylobaâ€™s extract against candida albicans. Iranian J Pharmacol Res 2006;4:285-9. |
[Table 1], [Table 2]