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Akbari Z, Dastan D, Fallah M, Maghsood A H, Matini M. Preliminary phytochemical screening and antitrichomonal activity of Ferula pseudalliacea. mljgoums 2025; 19 (6) :25-28
URL: http://mlj.goums.ac.ir/article-1-1771-en.html
URL: http://mlj.goums.ac.ir/article-1-1771-en.html
1- Department of Medical Parasitology and Mycology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
2- Department of Pharmacognosy, School of Pharmacy, Hamadan University of Medical Sciences, Hamadan, Iran; Medicinal Plants and Natural Products Research Center, Hamadan University of Medical Sciences, Hamadan, Iran
3- Department of Medical Parasitology and Mycology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran ,matini@umsha.ac.ir
2- Department of Pharmacognosy, School of Pharmacy, Hamadan University of Medical Sciences, Hamadan, Iran; Medicinal Plants and Natural Products Research Center, Hamadan University of Medical Sciences, Hamadan, Iran
3- Department of Medical Parasitology and Mycology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran ,
Keywords: Antitrichomonal agents, Essential oil, Phytochemicals, Trichomonas vaginalis, Ferula pseudalliacea
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Discussion
In this research, the antitrichomonal efficacy of F. pseudalliacea was evaluated in comparison with metronidazole, the first-line treatment for trichomoniasis. The results showed that F. pseudalliacea was potentially effective against T. vaginalis. The essential oil and n-hexane extract of F. pseudalliacea were the most potent antitrichomonal agents. After 48 hours of exposure, the MLC of these two potent products was 250 µg/ml. At sub-MLC concentrations, the GI% of the oil and the n-hexane extract was 92.8% and 50.6%, respectively.
To our knowledge, the antimicrobial activity of essential oil and crude extracts of F. pseudalliacea has not been previously investigated. However, the in vitro antiplasmodial and antibacterial activities of coumarin derivatives from F. pseudalliacea have been demonstrated. Anti-Plasmodium falciparum activity of sanandajin, methyl galbanate, and kamolonol acetate was reported with IC50 values of 2.6, 7.1, and 16.1 µM, respectively (9). Sanandajin and ethyl galbanate were effective against Staphylococcus aureus and Helicobacter pylori (MIC=64 μg/ml), and methyl galbanate was effective against a vancomycin-resistant strain of Enterococcus faecium (MIC=64 μg/ml) (12).
To date, many studies have explored the antimicrobial effects of medicinal plants, some specifically targeting the Trichomonas parasite. Twenty-six Iranian medicinal plants with antitrichomonal activity were reviewed by Ziaei Hezarjaribi et al. (18). In this review, Artemisia aucheri, Zataria multiflora, and Lavandula angustifolia were highlighted as the most potent medicinal plants. Other Iranian medicinal plants with strong antitrichomonal activity include Foeniculum vulgare, Marrubium vulgare, Pistacia atlantica subsp. kurdica, Plantago lanceolata L., and Ferula gummosa.
The antitrichomonal activity of F. vulgare was investigated by Karami et al. In their study, the methanolic and hexanic extracts of the medicinal herb showed the greatest potency with an MLC of 360 µg/ml. The essential oil demonstrated lower activity (MLC=1600 μg/ml). Chemical analysis of the essential oil revealed that E-anethole was the major component (19). In another study by Akbari et al., the essential oil of M. vulgare displayed the highest activity against T. vaginalis (MLC=291 μg/ml), while the n-hexane extract showed the lowest activity (MLC=1500 μg/ml) (20).
Matini and colleagues evaluated the efficacy of P. atlantica subsp. kurdica and P. lanceolata L. extracts against T. vaginalis. The ethyl acetate extract of these plants was considered the most potent antitrichomonal product, with MLC values of 337 and 1525 μg/ml, respectively (21,22). Anti-Trichomonas properties of various extracts of F. gummosa-including ethyl acetate, n-hexane, methanol, and its essential oil-were examined by Akbari et al. Their results showed that the extracts were more effective (MLC=125 μg/ml) than the essential oil (MLC=500 μg/ml). α- and β-Pinene, and β-eudesmol were identified as the major components of F. gummosa essential oil (23).
In another study, Mahmoudvand and colleagues examined the apoptotic effects of three Iranian herbs on T. vaginalis. They found that Quercus infectoria (IC50=3.4 µg/ml) was significantly more effective against the parasite than Satureja khuzestanica (IC50=5.1 µg/ml) and Pistacia khinjuk (IC50=26.6 µg/ml) (24).
Across studies reviewing the antitrichomonal potential of medicinal plants, the most active species typically belong to three families: Asteraceae, Lamiaceae, and Myrtaceae. Furthermore, terpenes, β-glycosides, saponins, essential oils, and alkaloids are among the major phytochemical compounds responsible for antitrichomonal activity (25). Berberine, an isoquinoline alkaloid used traditionally as a natural antibiotic, is one of the main components of Argemone mexicana methanolic extract. The IC50 antitrichomonal activity of A. mexicana extract was 70.8 and 67.2 μg/ml for the stem and leaf, respectively (26). Other plants with strong antitrichomonal effects include Persea americana, Verbascum thapsus, and Ocimum basilicum (27). The IC50 values of P. americana seed extracts against T. vaginalis were 0.524 and 0.533 µg/ml for the chloroform and ethanolic extracts, respectively (28). Alcoholic extract of V. thapsus and O. basilicum essential oil were effective against T. vaginalis at 30 and 39.17 µg/ml, respectively, after 24 hours of exposure (29,30).
In the present study, the observed antitrichomonal properties of F. pseudalliacea may be attributed to the presence of bioactive components. Preliminary analysis demonstrated that coumarins, terpenoids, and steroids were the major bioactive constituents in the extracts. Phytochemical screening analyses are helpful for identifying bioactive compounds and supporting the discovery and development of medicinal agents. These studies also enable quantitative and qualitative evaluation of active pharmaceutical compounds in crude extracts.
One of the advantages of this study was the use of several Trichomonas isolates, which increases the reliability of the results. Additionally, essential oil and extracts were examined simultaneously, allowing a more accurate comparison of their effects. However, one of the main limitations of this study was the lack of access to metronidazole-resistant Trichomonas isolates, preventing evaluation of the compounds’ effectiveness on resistant strains.
Conclusion
Despite limitations related to access to metronidazole-resistant isolates, F. pseudalliacea demonstrated potential anti-Trichomonas activity. The findings support the possibility of using components of F. pseudalliacea in the treatment of trichomoniasis. Therefore, further investigation into the bioactive components of F. pseudalliacea is recommended.
Acknowledgement
We would like to thank the staff of the Parasitology and Pharmacognosy Laboratories of Hamadan University of Medical Sciences for their cooperation in this project.
Funding sources
We are grateful to the Vice-Chancellor for Research and Technology, Hamadan University of Medical Sciences, for financial support of this research (Project No. 9409131443).
Ethical statement
This study was approved by the Ethical Committee of Hamadan University of Medical Sciences, Hamadan, Iran (UMSHA.REC.1394.84 IR).
Conflicts of interest
The authors declare that they have no conflict of interests to disclose.
Author contributions
DD, MF, AHM, and MM designed the study. ZA, DD, and MM conducted the experiment and data collection. MM and DD analyzed and interpreted the data and prepared the manuscript. All authors read and approved the final manuscript for publication.
Data availability statement
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Full-Text: (4 Views)
Introduction
Trichomoniasis is a common cause of human vaginitis caused by the protozoan parasite Trichomonas vaginalis. The infection is one of the most prevalent STIs worldwide. According to the World Health Organization in 2016, there were 156 million new cases of Trichomonas infection among persons aged 15 to 49 years. Trichomoniasis in women may present with a wide variety of clinical features, from an asymptomatic infection to severe vaginitis. Adverse pregnancy outcomes, infertility, and cervical neoplasia may be seen as complications after trichomoniasis. Furthermore, the infection increases the risk of spreading HIV infection in the community (1,2).
Metronidazole therapy is commonly the standard treatment for trichomoniasis worldwide. Since 1961, when metronidazole was introduced to treat trichomoniasis, it has faced challenges. One of these is drug resistance. Drug-resistant T. vaginalis is involved in an increasing number of refractory trichomoniasis cases. The first metronidazole-resistant trichomoniasis was reported in 1962, and it has been on the rise. According to the Center for Disease Control and Prevention, 2 to 5% of clinical T. vaginalis isolates are metronidazole-resistant in the United States (3,4).
Due to the presence of bioactive compounds in plants, they are of particular interest in pharmaceutical research. Plant essential oils and extracts are known to be a rich source of natural ingredients for the treatment of various diseases and are more compatible with biological systems. Ferula is a genus of plants with about 185 species. This genus has a special position in the Apiaceae family because of its pharmaceutical and industrial importance. In the Iranian flora, the genus Ferula contains 32 species, including 15 endemic plants, and this genus is typically called koma or kema. Several species of Ferula have been used in folk remedies as treatment of stomachache, hysteria, arthritis, rheumatoid disorders, and etc. Recent studies have proven antibacterial, antileishmanial, antimalarial, antioxidant, anti-epileptic, and anti-inflammatory effects of Ferula species (5-7). In previous studies, the methanol extract of F. szowitsiana was able to inhibit the growth of T. vaginalis cells (8).
F. pseudalliacea is an indigenous species of Iran and grows in the Sanandaj Mountains (West of Iran), and their gum has been used in traditional medicine for healing wounds and relieving itching. Recently, different studies have been done on the antibacterial, antiplasmodial, phytotoxic, and anticancer activity of F. pseudalliacea (9-12). But so far, a study to evaluate the antitrichomonas effect of this species has not been done.
In this study, we have examined the antitrichomonas effect of F. pseudalliacea essential oil and extracts against T. vaginalis. Also, the preliminary phytochemical analysis of the extracts was performed by standard methods.
Methods
Plant material
The roots of F. pseudalliacea were obtained from natural habitats in western Iran. The plant was identified in the herbarium of the Department of Pharmacognosy of Hamadan University of Medical Sciences, and voucher number 234 was allocated.
Preparation of the essential oil, extract, and phytochemical screening
The plant was dried using the shade-drying method at room temperature (20±5 °C), and the dried plant materials were crushed into powder. The powder (100 g) was used to prepare the essential oil using a Clevenger-type apparatus. The obtained essential oil was kept in an airtight container in a refrigerator (4 °C) until use. Extraction of the dried plant was performed using the maceration method. Briefly, the powdered plant (100 g) was macerated separately in n-hexane, ethyl acetate, and methanol solvents (3 × 2 L, room temperature for 72 h, 25 °C). Extraction was performed using a rotary evaporator below 40 °C. The obtained extracts were kept in dark containers in a refrigerator (4 °C) until use. Phytochemical analysis of the extracts was done using standard methods, and their constituent compounds were identified according to the method of Ugochukwu and Bargah (13,14).
Parasite culture and solutions
Five clinical T. vaginalis isolates were cultured in TYI-S-33 medium supplemented with 10% heat-inactivated adult bovine serum and antibiotics (100 IU/ml penicillin and 100 µg/ml streptomycin). After several 48-hour subcultures, pure trophozoites in the log phase of growth were used for the susceptibility assay (15,16). Metronidazole (Sigma-Aldrich, St Louis, USA) was dissolved in distilled water. The plant products were dissolved in dimethyl sulfoxide (D2650 SIGMA, BioReagent) or distilled water according to solubility. Solubility of the essential oil and the extracts in the culture medium was considered as the criterion for determining plant product concentrations, and susceptibility testing was started with the highest concentration. Solutions were prepared in 2-fold dilutions in culture medium for susceptibility assays at the following concentrations: 200, 100, 50, 25, 12.5, 6.2, 3.1, 1.6, 0.8, 0.4, 0.2, 0.1 µg/ml for metronidazole, and 4000, 2000, 1000, 500, 250, 125, 62.5 µg/ml for the plant products.
Susceptibility assay
The minimum lethal concentration (MLC) corresponds to the lowest concentration of the antitrichomonal agents that kills all trophozoites after exposure. Growth inhibition due to sub-MLC and lower concentrations of the agents was considered as the percentage of growth inhibition (GI% = a – b / a × 100; a = mean number of viable trophozoites in the negative control well, b = number of viable trophozoites in the test well at ≤ sub-MLC concentration). Susceptibility testing was assessed according to the method recommended by the CDC (17).
The experiments were performed in 96-well microtiter plates. First, the required serial dilutions of the agents were prepared, and 100 µL of the prepared solutions was dispensed into the wells. Then, the parasites in the logarithmic growth phase were counted with a hemocytometer (Neubauer cell chamber), and 100 µL of parasite-containing medium (2 × 10⁵ trophozoites/mL) was added to each test well. Finally, the number of Trichomonas cells was set to 2 × 10⁴ cells/well. The plates were aerobically incubated at 35.5 °C. After 24 and 48 hours of exposure, the test plates were examined with an inverted microscope to determine the MLC. The lowest concentration of the essential oil and extracts in the test well where no motile parasites were observed was considered the MLC concentration.
To evaluate the GI% rate of the agents, the number of parasites in the test wells was counted and compared with the number of parasites in the negative control well according to the equation mentioned above. The experiments were repeated in pairs and twice separately under sterile conditions. In each run, control wells (Negative control and metronidazole control) were used to check the experimental conditions. At the end, exposed parasites were cultured in fresh medium for MLC confirmation.
Statistical analysis
Analysis was performed using SPSS statistical software, version 16. The results were shown as MLC and mean values. The Friedman test was used to compare the averages. A P value less than 0.05 was considered statistically significant.
Results
Susceptibility testing revealed that the extracts and essential oil of F. pseudalliacea had a lethal effect on the Trichomonas parasite. The antitrichomonal activity of F. pseudalliacea depended on concentration and time of exposure, as shown in Tables 1 and 2. At MLC concentrations, the tested agents were able to kill all trophozoites, which was confirmed by culturing treated trophozoites in fresh medium. The essential oil of F. pseudalliacea exhibited the highest anti-Trichomonas potential with an MLC of 250 µg/ml, followed by the ethyl acetate and n-hexane extracts with MLC of 500 µg/ml, after 24 hours of incubation (P=0.002). After 48 hours, the antitrichomonal activity of the n-hexane extract increased to an MLC of 250 µg/ml (Table 2). At the sub-MLC concentrations, growth inhibition of the trichomonads was observed, and the number of live trophozoites was remarkably reduced compared to the control (Tables 1 and 2). GI% of the agents at the sub-MLC concentrations ranged from 35.2% to 87.0% after 24 hours, and from 42.8% to 92.8% after 48 hours. Drug susceptibility testing demonstrated that the Trichomonas isolates were susceptible to metronidazole, with MLCs ranging from 6.2 to 12.5 µg/ml (Table 3).
Preliminary phytochemical tests for extracts were studied by standard methods. The results showed F. pseudalliacea extracts contain bioactive constituents including coumarins, terpenoids, steroids, phenolics, tannins, and glycosides (Table 4). The major constituents were coumarins, terpenoids, and steroids.
Trichomoniasis is a common cause of human vaginitis caused by the protozoan parasite Trichomonas vaginalis. The infection is one of the most prevalent STIs worldwide. According to the World Health Organization in 2016, there were 156 million new cases of Trichomonas infection among persons aged 15 to 49 years. Trichomoniasis in women may present with a wide variety of clinical features, from an asymptomatic infection to severe vaginitis. Adverse pregnancy outcomes, infertility, and cervical neoplasia may be seen as complications after trichomoniasis. Furthermore, the infection increases the risk of spreading HIV infection in the community (1,2).
Metronidazole therapy is commonly the standard treatment for trichomoniasis worldwide. Since 1961, when metronidazole was introduced to treat trichomoniasis, it has faced challenges. One of these is drug resistance. Drug-resistant T. vaginalis is involved in an increasing number of refractory trichomoniasis cases. The first metronidazole-resistant trichomoniasis was reported in 1962, and it has been on the rise. According to the Center for Disease Control and Prevention, 2 to 5% of clinical T. vaginalis isolates are metronidazole-resistant in the United States (3,4).
Due to the presence of bioactive compounds in plants, they are of particular interest in pharmaceutical research. Plant essential oils and extracts are known to be a rich source of natural ingredients for the treatment of various diseases and are more compatible with biological systems. Ferula is a genus of plants with about 185 species. This genus has a special position in the Apiaceae family because of its pharmaceutical and industrial importance. In the Iranian flora, the genus Ferula contains 32 species, including 15 endemic plants, and this genus is typically called koma or kema. Several species of Ferula have been used in folk remedies as treatment of stomachache, hysteria, arthritis, rheumatoid disorders, and etc. Recent studies have proven antibacterial, antileishmanial, antimalarial, antioxidant, anti-epileptic, and anti-inflammatory effects of Ferula species (5-7). In previous studies, the methanol extract of F. szowitsiana was able to inhibit the growth of T. vaginalis cells (8).
F. pseudalliacea is an indigenous species of Iran and grows in the Sanandaj Mountains (West of Iran), and their gum has been used in traditional medicine for healing wounds and relieving itching. Recently, different studies have been done on the antibacterial, antiplasmodial, phytotoxic, and anticancer activity of F. pseudalliacea (9-12). But so far, a study to evaluate the antitrichomonas effect of this species has not been done.
In this study, we have examined the antitrichomonas effect of F. pseudalliacea essential oil and extracts against T. vaginalis. Also, the preliminary phytochemical analysis of the extracts was performed by standard methods.
Methods
Plant material
The roots of F. pseudalliacea were obtained from natural habitats in western Iran. The plant was identified in the herbarium of the Department of Pharmacognosy of Hamadan University of Medical Sciences, and voucher number 234 was allocated.
Preparation of the essential oil, extract, and phytochemical screening
The plant was dried using the shade-drying method at room temperature (20±5 °C), and the dried plant materials were crushed into powder. The powder (100 g) was used to prepare the essential oil using a Clevenger-type apparatus. The obtained essential oil was kept in an airtight container in a refrigerator (4 °C) until use. Extraction of the dried plant was performed using the maceration method. Briefly, the powdered plant (100 g) was macerated separately in n-hexane, ethyl acetate, and methanol solvents (3 × 2 L, room temperature for 72 h, 25 °C). Extraction was performed using a rotary evaporator below 40 °C. The obtained extracts were kept in dark containers in a refrigerator (4 °C) until use. Phytochemical analysis of the extracts was done using standard methods, and their constituent compounds were identified according to the method of Ugochukwu and Bargah (13,14).
Parasite culture and solutions
Five clinical T. vaginalis isolates were cultured in TYI-S-33 medium supplemented with 10% heat-inactivated adult bovine serum and antibiotics (100 IU/ml penicillin and 100 µg/ml streptomycin). After several 48-hour subcultures, pure trophozoites in the log phase of growth were used for the susceptibility assay (15,16). Metronidazole (Sigma-Aldrich, St Louis, USA) was dissolved in distilled water. The plant products were dissolved in dimethyl sulfoxide (D2650 SIGMA, BioReagent) or distilled water according to solubility. Solubility of the essential oil and the extracts in the culture medium was considered as the criterion for determining plant product concentrations, and susceptibility testing was started with the highest concentration. Solutions were prepared in 2-fold dilutions in culture medium for susceptibility assays at the following concentrations: 200, 100, 50, 25, 12.5, 6.2, 3.1, 1.6, 0.8, 0.4, 0.2, 0.1 µg/ml for metronidazole, and 4000, 2000, 1000, 500, 250, 125, 62.5 µg/ml for the plant products.
Susceptibility assay
The minimum lethal concentration (MLC) corresponds to the lowest concentration of the antitrichomonal agents that kills all trophozoites after exposure. Growth inhibition due to sub-MLC and lower concentrations of the agents was considered as the percentage of growth inhibition (GI% = a – b / a × 100; a = mean number of viable trophozoites in the negative control well, b = number of viable trophozoites in the test well at ≤ sub-MLC concentration). Susceptibility testing was assessed according to the method recommended by the CDC (17).
The experiments were performed in 96-well microtiter plates. First, the required serial dilutions of the agents were prepared, and 100 µL of the prepared solutions was dispensed into the wells. Then, the parasites in the logarithmic growth phase were counted with a hemocytometer (Neubauer cell chamber), and 100 µL of parasite-containing medium (2 × 10⁵ trophozoites/mL) was added to each test well. Finally, the number of Trichomonas cells was set to 2 × 10⁴ cells/well. The plates were aerobically incubated at 35.5 °C. After 24 and 48 hours of exposure, the test plates were examined with an inverted microscope to determine the MLC. The lowest concentration of the essential oil and extracts in the test well where no motile parasites were observed was considered the MLC concentration.
To evaluate the GI% rate of the agents, the number of parasites in the test wells was counted and compared with the number of parasites in the negative control well according to the equation mentioned above. The experiments were repeated in pairs and twice separately under sterile conditions. In each run, control wells (Negative control and metronidazole control) were used to check the experimental conditions. At the end, exposed parasites were cultured in fresh medium for MLC confirmation.
Statistical analysis
Analysis was performed using SPSS statistical software, version 16. The results were shown as MLC and mean values. The Friedman test was used to compare the averages. A P value less than 0.05 was considered statistically significant.
Results
Susceptibility testing revealed that the extracts and essential oil of F. pseudalliacea had a lethal effect on the Trichomonas parasite. The antitrichomonal activity of F. pseudalliacea depended on concentration and time of exposure, as shown in Tables 1 and 2. At MLC concentrations, the tested agents were able to kill all trophozoites, which was confirmed by culturing treated trophozoites in fresh medium. The essential oil of F. pseudalliacea exhibited the highest anti-Trichomonas potential with an MLC of 250 µg/ml, followed by the ethyl acetate and n-hexane extracts with MLC of 500 µg/ml, after 24 hours of incubation (P=0.002). After 48 hours, the antitrichomonal activity of the n-hexane extract increased to an MLC of 250 µg/ml (Table 2). At the sub-MLC concentrations, growth inhibition of the trichomonads was observed, and the number of live trophozoites was remarkably reduced compared to the control (Tables 1 and 2). GI% of the agents at the sub-MLC concentrations ranged from 35.2% to 87.0% after 24 hours, and from 42.8% to 92.8% after 48 hours. Drug susceptibility testing demonstrated that the Trichomonas isolates were susceptible to metronidazole, with MLCs ranging from 6.2 to 12.5 µg/ml (Table 3).
Preliminary phytochemical tests for extracts were studied by standard methods. The results showed F. pseudalliacea extracts contain bioactive constituents including coumarins, terpenoids, steroids, phenolics, tannins, and glycosides (Table 4). The major constituents were coumarins, terpenoids, and steroids.
|
Table 1. Efficacy of essential oil and extracts of F. pseudalliacea on T. vaginalis after 24 hours’ exposure
.PNG) a Minimum Lethal Concentration (MLC) is related to the lowest concentration of the antitrichomonal agents that kill all trichomonads b Sub-MLC concentration is related to inhibition of trichomonads growth Table 2. Efficacy of essential oil and extracts of F. pseudalliacea on T. vaginalis after 48 hours’ exposure .PNG) a Minimum Lethal Concentration (MLC) is related to the lowest concentration of the antitrichomonal agents that kill all trichomonads b Sub-MLC concentration is related to inhibition of trichomonads growth Table 3. Efficacy of metronidazole on T. vaginalis .PNG) a Minimum Lethal Concentration (MLC) is related to the lowest concentration of the antitrichomonal agents that kill all trichomonads b Sub-MLC concentration is related to inhibition of trichomonads growth |
|
Table 4. Preliminary phytochemical screening of F. pseudalliacea extracts
.PNG) + Presence; - Absence |
Discussion
In this research, the antitrichomonal efficacy of F. pseudalliacea was evaluated in comparison with metronidazole, the first-line treatment for trichomoniasis. The results showed that F. pseudalliacea was potentially effective against T. vaginalis. The essential oil and n-hexane extract of F. pseudalliacea were the most potent antitrichomonal agents. After 48 hours of exposure, the MLC of these two potent products was 250 µg/ml. At sub-MLC concentrations, the GI% of the oil and the n-hexane extract was 92.8% and 50.6%, respectively.
To our knowledge, the antimicrobial activity of essential oil and crude extracts of F. pseudalliacea has not been previously investigated. However, the in vitro antiplasmodial and antibacterial activities of coumarin derivatives from F. pseudalliacea have been demonstrated. Anti-Plasmodium falciparum activity of sanandajin, methyl galbanate, and kamolonol acetate was reported with IC50 values of 2.6, 7.1, and 16.1 µM, respectively (9). Sanandajin and ethyl galbanate were effective against Staphylococcus aureus and Helicobacter pylori (MIC=64 μg/ml), and methyl galbanate was effective against a vancomycin-resistant strain of Enterococcus faecium (MIC=64 μg/ml) (12).
To date, many studies have explored the antimicrobial effects of medicinal plants, some specifically targeting the Trichomonas parasite. Twenty-six Iranian medicinal plants with antitrichomonal activity were reviewed by Ziaei Hezarjaribi et al. (18). In this review, Artemisia aucheri, Zataria multiflora, and Lavandula angustifolia were highlighted as the most potent medicinal plants. Other Iranian medicinal plants with strong antitrichomonal activity include Foeniculum vulgare, Marrubium vulgare, Pistacia atlantica subsp. kurdica, Plantago lanceolata L., and Ferula gummosa.
The antitrichomonal activity of F. vulgare was investigated by Karami et al. In their study, the methanolic and hexanic extracts of the medicinal herb showed the greatest potency with an MLC of 360 µg/ml. The essential oil demonstrated lower activity (MLC=1600 μg/ml). Chemical analysis of the essential oil revealed that E-anethole was the major component (19). In another study by Akbari et al., the essential oil of M. vulgare displayed the highest activity against T. vaginalis (MLC=291 μg/ml), while the n-hexane extract showed the lowest activity (MLC=1500 μg/ml) (20).
Matini and colleagues evaluated the efficacy of P. atlantica subsp. kurdica and P. lanceolata L. extracts against T. vaginalis. The ethyl acetate extract of these plants was considered the most potent antitrichomonal product, with MLC values of 337 and 1525 μg/ml, respectively (21,22). Anti-Trichomonas properties of various extracts of F. gummosa-including ethyl acetate, n-hexane, methanol, and its essential oil-were examined by Akbari et al. Their results showed that the extracts were more effective (MLC=125 μg/ml) than the essential oil (MLC=500 μg/ml). α- and β-Pinene, and β-eudesmol were identified as the major components of F. gummosa essential oil (23).
In another study, Mahmoudvand and colleagues examined the apoptotic effects of three Iranian herbs on T. vaginalis. They found that Quercus infectoria (IC50=3.4 µg/ml) was significantly more effective against the parasite than Satureja khuzestanica (IC50=5.1 µg/ml) and Pistacia khinjuk (IC50=26.6 µg/ml) (24).
Across studies reviewing the antitrichomonal potential of medicinal plants, the most active species typically belong to three families: Asteraceae, Lamiaceae, and Myrtaceae. Furthermore, terpenes, β-glycosides, saponins, essential oils, and alkaloids are among the major phytochemical compounds responsible for antitrichomonal activity (25). Berberine, an isoquinoline alkaloid used traditionally as a natural antibiotic, is one of the main components of Argemone mexicana methanolic extract. The IC50 antitrichomonal activity of A. mexicana extract was 70.8 and 67.2 μg/ml for the stem and leaf, respectively (26). Other plants with strong antitrichomonal effects include Persea americana, Verbascum thapsus, and Ocimum basilicum (27). The IC50 values of P. americana seed extracts against T. vaginalis were 0.524 and 0.533 µg/ml for the chloroform and ethanolic extracts, respectively (28). Alcoholic extract of V. thapsus and O. basilicum essential oil were effective against T. vaginalis at 30 and 39.17 µg/ml, respectively, after 24 hours of exposure (29,30).
In the present study, the observed antitrichomonal properties of F. pseudalliacea may be attributed to the presence of bioactive components. Preliminary analysis demonstrated that coumarins, terpenoids, and steroids were the major bioactive constituents in the extracts. Phytochemical screening analyses are helpful for identifying bioactive compounds and supporting the discovery and development of medicinal agents. These studies also enable quantitative and qualitative evaluation of active pharmaceutical compounds in crude extracts.
One of the advantages of this study was the use of several Trichomonas isolates, which increases the reliability of the results. Additionally, essential oil and extracts were examined simultaneously, allowing a more accurate comparison of their effects. However, one of the main limitations of this study was the lack of access to metronidazole-resistant Trichomonas isolates, preventing evaluation of the compounds’ effectiveness on resistant strains.
Conclusion
Despite limitations related to access to metronidazole-resistant isolates, F. pseudalliacea demonstrated potential anti-Trichomonas activity. The findings support the possibility of using components of F. pseudalliacea in the treatment of trichomoniasis. Therefore, further investigation into the bioactive components of F. pseudalliacea is recommended.
Acknowledgement
We would like to thank the staff of the Parasitology and Pharmacognosy Laboratories of Hamadan University of Medical Sciences for their cooperation in this project.
Funding sources
We are grateful to the Vice-Chancellor for Research and Technology, Hamadan University of Medical Sciences, for financial support of this research (Project No. 9409131443).
Ethical statement
This study was approved by the Ethical Committee of Hamadan University of Medical Sciences, Hamadan, Iran (UMSHA.REC.1394.84 IR).
Conflicts of interest
The authors declare that they have no conflict of interests to disclose.
Author contributions
DD, MF, AHM, and MM designed the study. ZA, DD, and MM conducted the experiment and data collection. MM and DD analyzed and interpreted the data and prepared the manuscript. All authors read and approved the final manuscript for publication.
Data availability statement
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Research Article: Research Article |
Subject:
Parasitology
Received: 2024/01/7 | Accepted: 2024/04/20 | Published: 2025/12/27 | ePublished: 2025/12/27
Received: 2024/01/7 | Accepted: 2024/04/20 | Published: 2025/12/27 | ePublished: 2025/12/27
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