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Abstract
In this study, we have evaluated the effect of essential oils of Thymus broussonetii Boiss, an endemic plant of Morocco in experimental transmission of Toxoplasma gondii cysts in mice. These oils were obtained by hydrodistillation and were administered to mice at 20 microg/animal orally at the time of infection and for several days thereafter. This resulted in total absence of intracerebral cysts in mice who received the essential oils signifying that these essential oils of thyme have a blocking effect on the appearance of the cysts. In addition, no abnormality was observed in the control mice who received the essential oils of thyme.
... e anticancer properties of T. broussonetii EOs have also been reported against various tumor cell lines like P815 mastocytoma, CEM, and K-562 [12,15,21]. Moreover, T. broussonetii was revealed to exhibit anti-inflammatory activity [28], anticorrosive potential [23], insecticidal [19,27,29], antiparasitic [30], antipyretic [22], antinociceptive [31], immunological, and behavioral effects [31]. In addition, the acute toxicological investigations of T. broussonetii EOs have shown death cases and some signs of toxicity [22]. ...
... In mice given the essential oils, no cyst was observed. In addition, no disorder was noted in the control animals given the thyme EOs [30] ( Table 6). ...
Thymus broussonetii Boiss (T. broussonetii) is a rare medicinal and aromatic plant. It is widely used in traditional medicine to treat several diseases, including diarrhea, fever, cough, irritation, skin diseases, rheumatism, respiratory ailments, influenza, and digestion problems. In this review, we have critically summarized previous data on T. broussonetii about its phytochemistry, botanical and geographical distribution, toxicological investigation, and pharmacological properties. Using scientific research databases such as Wiley Online, SciFinder, ScienceDirect, PubMed, SpringerLink, Web of Science, Scopus Wiley Online, and Google Scholar, the data on T. broussonetii were collected and discussed. The presented data regrouped bioactive compounds and biological activities of T. broussonetii. The findings of this work showed that essential oils and extracts of T. broussonetii exhibited numerous pharmacological activities (in vitro and in vivo), particularly antibacterial, antifungal, antioxidant, anticancer, anti-inflammatory, insecticidal, antipyretic, antinociceptive, and immunological and behavioral effects. While toxicological studies of T. broussonetii essential oils and extracts are lacking, modern scientific tools revealed the presence of different classes of secondary metabolites such as terpenoids, alkaloids, flavonoids, tannins, coumarins, quinones, carotenoids, and steroids. T. broussonetii essential oils, especially from the aerial parts, exhibited potent antibacterial, antifungal, and antioxidant effects. An in-depth toxicological investigation is needed to validate the efficacy and safety of T. broussonetii extracts and essential oils and their secondary metabolites. However, further pharmacokinetic and pharmacodynamic studies should be performed to validate its bioavailability.
... Thymus is one of the variety of herbs used in cooking and medicines, which is known as an antimicrobial, antispasmodic, anti-inflammatory and antioxidant medicine. Thymus genus has several species, such as T. broussontii, which is used in the treatment of diarrhea and fever [39]. Previous data have indicated that, use of the essential oil of T. broussonetii Boiss prevented the development of cystic shapes of T. gondii in the mice. ...
... Previous data have indicated that, use of the essential oil of T. broussonetii Boiss prevented the development of cystic shapes of T. gondii in the mice. In this study, in the rat groups that were initially infected with cystic shapes and, then, received T. hymus broussonetii, the cystic shapes in the brain were not seen in the days after the infection of the mice (OF1), which could be due to the inhibitory role of essential oils in the destruction of prescriptive cysts and the release of bradyzoites in the intestines of the mice [39]. ...
Background
Toxoplasma gondii is the global protozoa that could cause contamination in warm-blooded animals and is considered among the opportunistic pathogens in immunocompromised patients. Among the people at risk, toxoplasmosis infection can lead to the incidence of severe clinical manifestations, encephalitis, chorioretinitis, and even death.PurposeThe present research is focused on the new research for the treatment of toxoplasmosis parasitic disease using medicinal herbs.Methods
The search was performed in five English databases, including Scopus, PubMed, Web of Science, EMBASE, and Google Scholar up from 2010 to December 2019. Studies in any language were entered in the searching step if they had an English abstract. The words and terms were used as a syntax with specific tags of each database.ResultsOut of 1832 studies, 36 were eligible to be reviewed. The findings showed that 17 studies (47%) were performed in vitro, 14 studies (39%) in vivo, and 5 studies (14%) both in vivo and in vitro.Conclusion
The studies showed that the plant extracts can be a good alternative in reducing the toxoplasmosis effects in the host and the herbal extracts can be used to produce natural product-based drugs affecting toxoplasmosis with fewer side-effects than synthetic drugs.
... Several of them are used in traditional phytotherapy in the form of tisane or powder as an antimicrobial, antispasmodic, anti-inflammatory, antioxidant, and antifungal agent [2][3][4][5][6][7] . These biological activities are related to the presence of phenolic compounds in the extracts such as thymol and/or carvacrol and other compounds [8][9][10] . In the Moroccan flora, Thymus broussonetii Boiss. is an endemic species [11] which present a great economic importance [12] . ...
... It is also used as a tonifying agent [12,13] . Recently, the antiparasitic effect (against Toxoplasma gondii cysts) of Thymus broussonetii essential oil was found [9] . It is reported that extracts of T. broussonetii possess not only anti-inflammatory effects [14] , but also stimulatory effects on the immune system in addition to protection against stress by a neurotropic and antinociceptive activity [15] . ...
This paper reports, for the first time, a rapid and reliable micropropagation protocol
for high-frequency shoot regeneration and plant establishment of Thymus broussonetii, a
vulnerable and highly valuable aromatic and medicinal plant. This plant species is threatened
by over collection due to its socioeconomic importance. Hence, an attempt has been made for
in vitro cultivation of this plant for large scale multiplication as well as conservation. Several
treatments such as cytokinin types and concentrations, cytokinin/auxin ratio, gelling agents,
and explant density were tested on the shoot bud induction. Effects of various concentrations
of auxins were also examined on the micropropagated shoots during rooting stage. Fullstrength
Murashige and Skoog (MS) medium supplemented with 6- benzylaminopurine (4.4
μM) and naphthalene acetic acid (1 μM) promoted significantly shoot bud induction while
affecting shoot length. The use of 3 g/l of gellan gum (Phytagel™) promoted shoot bud
induction and growth. For micropropagated shoot rooting, hormone-free MS medium
exhibited the best results. The micropropagated plantlets were successfully hardened and 94%
of them were found healthy after transplantation to ex vitro conditions. The micropropagated
plants established in soil showed normal flowering did not show any morphological
abnormality during one year of growth.
... The action of Thymus broussonetii (thyme) essential oil, rich in carvacrol (83.18%), on animals infected with T. gondii (PRU strain) were observed by Dahbi et al. (2010). For fifteen days, mice were treated with essential oil and then euthanized. ...
Awareness about pollution worldwide through the excessive use of chemicals that affect every part of our environment makes the urgent need to use natural alternatives eco-friendly materials to reduce the losses come from synthetic chemicals. In recent decades, a great leap has been made to control weed by synthetic herbicides which are considered the most effective weed control method in comparison with the other methods. Although synthetic herbicides are considered less toxic between other pesticides such as insecticides and fungicides, the usual use, even if it is used at the recommended rates harmed the environment and human health. Previous works indicated that Essential oils have been demonstrated to have good phytotoxic activity on various plant species by suppressing germination and reducing growth parameters. Phytotoxic activity of essential oils can act directly as bioherbicide by effects on one or more than one of the biological processes inside plants which cause the death of plant completely or partially. This chapter highlights the desirable phytotoxic activity of essential oils and their possible uses as natural weed killers.KeywordsAllelopathyEssential oilsPhytotoxic activitySynthetic herbicidesNatural weed killer
... The action of Thymus broussonetii (thyme) essential oil, rich in carvacrol (83.18%), on animals infected with T. gondii (PRU strain) were observed by Dahbi et al. (2010). For fifteen days, mice were treated with essential oil and then euthanized. ...
Fungi are organisms that feed from organic matter and act as parasites in living organisms. They are found in different environments and their growth in food products has become a global concern, as well as the resistance to commercial fungicides, due to the economic damage generated by food deterioration and the harmful effects on human health. Thus, the search for new natural fungicides has increased, such as essential oils from plants that have been promising in combating fungi, improving the quantity and quality of foods with low toxicity. This chapter aimed to carry out a bibliographical review on promising essential oils in combating fungal species of the genera Aspergillus, Penicillium, Fusarium, Alternaria, Candida and Cladosporium, the main food spoilage, and their constituents.KeywordsFungiFoodDeteriorationEssential oil
Aspergillus
Penicillium
Fusarium
Alternaria
Candida
Cladosporium
... The action of Thymus broussonetii (thyme) essential oil, rich in carvacrol (83.18%), on animals infected with T. gondii (PRU strain) were observed by Dahbi et al. (2010). For fifteen days, mice were treated with essential oil and then euthanized. ...
Toxoplasmosis is a serious disease caused by Toxoplasma gondii. Natural products have been reported in the literature as an effective therapeutic alternative in the treatment of this infection. Among the natural products, we highlight the use of essential oils that have numerous properties in the treatment of various diseases, including anti-T. gondii. In this chapter, we discuss the effect of essential oils, the majority of them obtained from plants used as food ingredient, on the parasite that causes toxoplasmosis, their mechanisms of action, the synergistic effects between essential oil components, as well as the importance of using encapsulations. The phytochemical compounds are said to be responsible for the anti-T. gondii activity of essential oils and are therefore considered an excellent alternative for the search of new compounds for the treatment of toxoplasmosis. Further research will provide better understandings of the potential of essential oils against T. gondii, thus aiding in the search for therapeutically active solutions from natural products.
... The action of Thymus broussonetii (thyme) essential oil, rich in carvacrol (83.18%), on animals infected with T. gondii (PRU strain) were observed by Dahbi et al. (2010). For fifteen days, mice were treated with essential oil and then euthanized. ...
Essential oils are chemical substances composed mainly of terpenes and terpenoids. These compounds are widely investigated due to their lipophilic characteristics and biological and pharmacological properties. Therefore, the objective of this review was to investigate the potential of essential oils against species of the genus Leishmania, parasites causing the infectious disease leishmaniasis. Using the descriptors “Essential oil AND Leishmania” information available in the Scopus© database was collected. 114 articles met the inclusion criteria and were selected for analysis in this study. Among the tests performed, most were of the in vitro type (97.4%) and L. amazonensis (47.4%) and L. infantum (28.9%) were the species most used in these studies. Among the studies that investigated the mechanism of action, the essential oil of the species Tetradenia riparia showed the best result of IC50 (0.03μg/mL) against L. amazonensis isolates. Several works attribute the anti-Leishmania activities of essential oils with different bioactivities, such as: morphological and immunological alterations, antioxidant capacity and enzymatic action. Finally, it is concluded that essential oils have great potential for the development of new drugs against leishmaniasis, however further research is needed through in vivo tests to elucidate the mechanisms of action of such compounds.KeywordsLeishamaniasisTerpenesTerpernoidsMedicinal plantAnthropozoonosisMonoterpenesSesquiterpenes
Lutzomyia
Phlebotomus
Mechanisms of action
... Mice infected with the type II Prugniaud (PRU) T. gondii strain developed intracerebral cysts after 3 weeks (91). However, animals treated with the oil extract did not exhibit brain cysts (91). In another study, ethanolic extract from the leaves and ariel parts of Thymus vulgaris reduced the severity of chronic infection. ...
Toxoplasma gondii is an obligate intracellular parasite that mainly infects warm-blooded animals including humans. T. gondii can encyst and persist chronically in the brain, leading to a broad spectrum of neurological sequelae. Despite the associated health threats, no clinical drug is currently available to eliminate T. gondii cysts. In a continuous effort to uncover novel therapeutic agents for these cysts, the potential of nutritional products has been explored. Herein, we describe findings from in vitro and in vivo studies that support the efficacy of plant-based foods and nutraceuticals against brain cyst burden and cerebral pathologies associated with chronic toxoplasmosis. Finally, we discuss strategies to increase the translatability of preclinical studies and nutritional products to address whether nutritional therapy can be beneficial for coping with chronic T. gondii infections in humans.
... Fluorescence area indicates the growth of T. gondii during different treatment. (A-D) Different concentrations of Ov EO, (A) 70 mg/ml, (B) 35 mg/ml, (C) 17 mg/ml, (D) 9 mg/ml, and (E) no treatment; (F-H) Different concentrations of Ca, (F) 17 mg/ml, (G) 9 mg/ml, (H) 4 mg/ml, (I) SMZ, and (J) DMSO.essential oils had the inhibitory effect on T. gondii, in which, Bunium persicum (Boiss) EO(Tavakoli Kareshk et al., 2015) and Thymus broussonetii Boiss EO(Dahbi et al., 2010) played obvious roles in acute and chronic T. gondii infection in mice respectively. Ov EO is an important condiment, perfume, cosmetics, incense and preservative, with a long history of application(Diniz do Nascimento et al., 2020). ...
Toxoplasma gondii is a serious hazard to public health and animal husbandry. Due to the current dilemma of treatment of toxoplasmosis, it is urgent to find new anti- T. gondii drugs to treat toxoplasmosis. In this study, the anti- T. gondii activity of Origanum vulgare essential oil ( Ov EO) was firstly studied, and then, carvanol (Ca), the main ingredient of Ov EO was evaluated using the MTT assay on human foreskin fibroblast (HFF) cells in vitro . The cytotoxicity was evaluated using the MTT assay on HFF cells. The CC 50 of Ov EO and Ca was 134.9 and 43.93 μg/ml, respectively. Both of them exhibited anti-parasitic activity, and inhibited the growth of T. gondii in a dose-dependent manner. For the inhibition effect, Ca was better than Ov EO at the same concentration, the IC 50 of Ov EO and Ca was 16.08 and 7.688 μg/ml, respectively. In addition, treatment with Ca, was found to change the morphology of T. gondii tachyzoites and made their shapes curl up. These results showed that Ca was able to inhibit the proliferation of T. gondii by reducing invasion, which may be due to its detrimental effect on the mobility of tachyzoites. Our results indicated that Ca could be a potential new and effective drug for treating toxoplasmosis.
... The results showed a reduction in tissue cysts in the brain. The mechanism of action could be due to the lysis of the bradyzoites released in the intestine by the action of the essential oil components [42]. ...
Toxoplasma gondii, an apicomplexan protozoan that is considered an opportunistic parasite of medical and veterinary interest, causes toxoplasmosis, which may be asymptomatic in the immunocompetent host, while fatal in the immunocompromised patient. A combination of pyrimethamine-sulfadiazine is the treatment of choice; nevertheless, these two drugs produce severe side effects and are only effective against acute toxoplasmosis, hence, less toxic novel compounds with anti-toxoplasma activity are greatly needed. Natural compounds seem to be a promising source to identify lead compounds against T. gondii. In this review, the in vitro and in vivo activities of extracts, fractions, and isolated compounds obtained from different plants are described. In addition, some biological and pathological generalities of the parasite are reviewed as well. Data were obtained from a bibliographic search throughout digital faculty libraries, Google Scholar search engine, Science Direct, SciELO databases, and the National Center for Biotechnology Information (NCBI). Founded records include the evaluation of 58 extracts, 14 fractions, and 7 compounds belonging to 53 species of 33 plant families. Predominant studies were made on in vivo RH T. gondii tachyzoites strain and very few in in vivo models of both acute and chronic toxoplasmosis. Research of natural compounds against T. gondii deserves more attention in order to identify novel drugs useful in the control of toxoplasmosis.
... En el grupo infectado y sin tratamiento se observaron entre 90 y 180 quistes tisulares de cerebro en comparación a cero quistes observados en los animales del grupo tratado. En este estudio se concluyó que la administración oral del aceite esencial de T. broroussonetii previene el desarrollo de quistes tisulares de T. gondii in vivo (Dahbi et al., 2010). ...
The apicomplex parasite Toxoplasma gondii (T. gondii) causes toxoplasmosis in humans. Currently pyrimethamine and sulfadiazine are the drugs of choice to treat the disease. However, produce serious adverse effects and result in unsuccessful treatments due to drug resistance. Thus, new effective compounds are needed to treat this infection and natural products can be a good source to obtain them, medicinal plants have been used to control other apicomplexan parasites. Pleopeltis crassinervata (P. crassinervata) is a fern used in some rural areas of Mexico to treat, among other ailments, mouth ulcers, gastrointestinal problems and parasites. In previous investigations it was demonstrated that P. crassinervata has activity against Trichomonas vaginalis (T. vaginalis). Due to their ethnobotanical records, the efficacy of the extracts, fractions and isolated subfractions obtained from the P. crassinervata fronds was evaluated on the viability of T. gondii tachyzoites RH strain using the Sytox Green method. Cytotoxicity of fractions and active subfractions was determined in the HEp-2, Vero and SH-SY5Y cell lines. Hexane methanolic fraction exhibited the highest antitoxoplasmic activity with an IC50 of 16.90 mg/mL, which was not toxic to host cells at doses up to 50 mg/mL. Subfractions named FH1 and FH5 isolated from hexane fraction were the most active of the subfractions evaluated against viability of T. gondii with IC50 values of 23.69 and 28.69 μg/mL respectively, which were not cytotoxic in concentrations up to 100 μg/mL. Lead compounds isolated from P. crassinervata are interesting to continue with their preclinical evaluation to characterize the active compound and determine the action mechanism.
... EOs that were extracted from several plant species have been reported to exhibit activity against T. gondii. EO from Thymus broussonetii Boiss, has been shown to reduce prugniaud (Pru) cysts (Dahbi et al., 2010). Bunium persicum (Boiss) EO has been shown to prevent T. gondii infection in mice and significantly delay their time of death (Kareshk et al., 2015). ...
Toxoplasmosis is a global zoonotic disease, and one-third of the human population is chronically infected by Toxoplasma gondii. Due to the limited effectiveness and prominent side effects of the existing drugs, there is a dire need for the discovery of new therapeutic options in the treatment of toxoplasmosis. In this study, five essential oils (EO) were screened for their anti-parasitic activity against T. gondii. The cytotoxicity of essential oils was evaluated using the MTT assay on human foreskin fibroblast cells. The CC50 values of Eucalyptus globulus EO, Cupressus sempervirens EO, Citrus aurantifolia EO, Melaleuca alternifolia EO, and Pelargonium X. asperum (Pa) EO were found to be 22.74, 7.25, 15.01, 6.26, and 4.77 mg/mL, respectively. Only PaEO exhibited anti-parasitic activity, and inhibited the growth of T. gondii in a dose-dependent manner. In addition, treatment with PaEO, was found to reduce the volume of T. gondii tachyzoites and make their membrane surfaces rough. These results showed that PaEO was able to inhibit the growth of T. gondii by reducing invasion, which may be due to its detrimental effect on the ability of tachyzoites to move. These findings suggest that PaEO could be a potential anti-T. gondii drug, which may facilitate the development of new and effective treatments against toxoplasmosis.
... However, the current first-line drugs can only suppress the replication of tachyzoites and are ineffective against the bradyzoite stage. Emerging studies have identified several natural compounds that can negate the brain cyst burden in murine toxoplasmosis (Dahbi et al., 2010;Schultz et al., 2014;Doggett et al., 2012;Bottari et al., 2015aBottari et al., , 2015bBottari et al., , 2016Eraky et al., 2016;Azami et al., 2018;Spalenka et al., 2020). Here, our study identifies UA as a novel compound that can interfere with cyst formation in-vitro and in-vivo. ...
Neurotoxoplasmosis, also known as cerebral toxoplasmosis, is an opportunistic chronic infection caused by the persistence of parasite Toxoplasma gondii in the latent cystic form in the brain. In wild animals, chronic infection is associated with behavioral manipulation evident by an altered risk perception towards predators. In humans, reactivation of cysts and conversion of quiescent parasites into highly invasive tachyzoites is a significant cause of mortality in immunocompromised patients. However, the current standard therapy for toxoplasmosis is not well tolerated and is ineffective against the parasite cysts. In recent years, the concept of dietary supplementation with products derived from plants has gained popularity as a natural remedy for brain disorders. Notably, urolithin-A, a metabolite produced in the gut following consumption of ellagitannins-enriched foods such as pomegranate, is reported to be blood-brain barrier permeable and exhibits neuroprotective effects in-vivo. In this study, we investigated the potential of pomegranate extract and urolithin-A as anti-neurotoxoplasmosis agents in-vitro and in-vivo. Treatment with pomegranate extract and urolithin-A reduced the parasite tachyzoites load and interfered with cysts development in differentiated human neural culture. Administration of urolithin-A also resulted in the formation of smaller brain cysts in chronically infected mice. Interestingly, this phenomenon was mirrored by an enhanced risk perception in the infected mice towards predatory cues. Together, our findings demonstrate the potential of dietary supplementation with urolithin-A enriched food as a novel natural remedy for the treatment of acute and chronic neurotoxoplasmosis.
... Benefits of the essential oils to decrease tissue cysts of T. gondii have been documented. 70 In this study, results from the acute-phase experiments revealed that the mean survival time in NE-no CR-treated mice was longer than that in negative control group. Moreover, the mean count of peritoneum tachyzoites in treatment mice was lower than that in control mice. ...
Background: The aim of this study was to prepare curcumin nanoemulsion (CR-NE) to solve the problems associated with poor water solubility and low bioavailability of CR and to test its efficiency in the treatment of acute and chronic toxoplasmosis in mouse models.
Materials and methods: CR-NE 1% was prepared using spontaneous emulsification by soybean as oil phase; a mixture of Tween 80 and Tween 85 as surfactant; ethanol as cosurfactant and distilled water. Particle size and zeta potential of NE were assessed using Nano-ZS90
dynamic light scattering. Stability testing of NE was assessed after storage for 2 months at room temperature. In vivo experiments were carried out using 50 BALB/c mice inoculated with virulent RH strain (type I) and 50 BALB/c mice inoculated with avirulent Tehran strain (type II) of Toxoplasma gondii and treated with CR-NE (1% w/v), CR suspension (CR-S, 1% w/v), and NE without CR (NE-no CR).
Results: The mean particle size and zeta potential of CR-NE included 215.66±16.8 nm and −29.46±2.65 mV, respectively, and were stable in particle size after a three freeze–thaw cycle. In acute phase experiment, the survival time of mice infected with RH strain of T. gondii and treated with CR-NE extended from 8 to 10 days post inoculation. The differences were statistically significant between the survival time of mice in CR-NE-treated group compared with negative
control group (P,0.001). Furthermore, CR-NE significantly decreased the mean counts of peritoneum tachyzoites from 5,962.5±666 in negative control group to 627.5±73 in CR-NE-treated mice (P=0.001). Growth inhibition rates of tachyzoites in peritoneum of mice receiving CR-NE, CR-S, and NE-no CR included 90%, 21%, and 11%, respectively, compared with negative control group. In chronic phase experiment, the average number and size of tissue cysts significantly decreased to 17.2±15.6 and 31.5±6.26 µm, respectively, in mice inoculated with bradyzoites of T. gondii Tehran strain and treated with CR-NE compared with that in negative control group (P,0.001). Decrease of cyst numbers was verified by downregulation of BAG1 in treatment groups compared with negative control group with a minimum relative expression in CR-NE (1.12±0.28), CR-S (11.76±0.87), and NE-no CR (14.67±0.77), respectively, (P,0.001).
Conclusion: Results from the current study showed the potential of CR-S and CR-NE in treatment of acute and chronic toxoplasmosis in mouse models for the first time. However, CR-NE was more efficient than CR-S, and it seems that CR-NE has a potential formula for the treatment of acute and chronic toxoplasmosis, especially in those with latent bradyzoites in brain.
... Benefits of the essential oils to decrease tissue cysts of T. gondii have been documented. 70 In this study, results from the acute-phase experiments revealed that the mean survival time in NE-no CR-treated mice was longer than that in negative control group. Moreover, the mean count of peritoneum tachyzoites in treatment mice was lower than that in control mice. ...
Background: The aim of this study was to prepare curcumin nanoemulsion (CR-NE) to solve the problems associated with poor water solubility and low bioavailability of CR and to test its efficiency in the treatment of acute and chronic toxoplasmosis in mouse models.Materials and methods: CR-NE 1% was prepared using spontaneous emulsification by soybean as oil phase; a mixture of Tween 80 and Tween 85 as surfactant; ethanol as cosurfactant
and distilled water. Particle size and zeta potential of NE were assessed using Nano-ZS90
dynamic light scattering. Stability testing of NE was assessed after storage for 2 months at room temperature. In vivo experiments were carried out using 50 BALB/c mice inoculated
with virulent RH strain (type I) and 50 BALB/c mice inoculated with avirulent Tehran strain
(type II) of Toxoplasma gondii and treated with CR-NE (1% w/v), CR suspension (CR-S, 1%
w/v), and NE without CR (NE-no CR).
Results: The mean particle size and zeta potential of CR-NE included 215.66±16.8 nm
and −29.46±2.65 mV, respectively, and were stable in particle size after a three freeze–thaw cycle.
In acute phase experiment, the survival time of mice infected with RH strain of T. gondii and
treated with CR-NE extended from 8 to 10 days postinoculation. The differences were statistically
significant between the survival time of mice in CR-NE-treated group compared with negative
control group (P,0.001). Furthermore, CR-NE significantly decreased the mean counts of peritoneum
tachyzoites from 5,962.5±666 in negative control group to 627.5±73 in CR-NE-treated
mice (P,0.001). Growth inhibition rates of tachyzoites in peritoneum of mice receiving CR-NE,
CR-S, and NE-no CR included 90%, 21%, and 11%, respectively, compared with negative control
group. In chronic phase experiment, the average number and size of tissue cysts significantly
decreased to 17.2±15.6 and 31.5±6.26 µm, respectively, in mice inoculated with bradyzoites of
T. gondii Tehran strain and treated with CR-NE compared with that in negative control group
(P,0.001). Decrease of cyst numbers was verified by downregulation of BAG1 in treatment
groups compared with negative control group with a minimum relative expression in CR-NE
(1.12±0.28), CR-S (11.76±0.87), and NE-no CR (14.67±0.77), respectively, (P,0.001).
Conclusion: Results from the current study showed the potential of CR-S and CR-NE in treatment
of acute and chronic toxoplasmosis in mouse models for the first time. However, CR-NE
was more efficient than CR-S, and it seems that CR-NE has a potential formula for the treatment
of acute and chronic toxoplasmosis, especially in those with latent bradyzoites in brain.
Keywords: curcumin nanoemulsion, curcumin suspension, Toxoplasma gondii RH strain,
Toxoplasma gondii Tehran strain, soybean oil, blood–brain barrier, spontaneous emulsification
... Benefits of the essential oils to decrease tissue cysts of T. gondii have been documented. 70 In this study, results from the acute-phase experiments revealed that the mean survival time in NE-no CR-treated mice was longer than that in negative control group. Moreover, the mean count of peritoneum tachyzoites in treatment mice was lower than that in control mice. ...
Background: The aim of this study was to prepare curcumin nanoemulsion (CR-NE) to solve the problems associated with poor water solubility and low bioavailability of CR and to test its efficiency in the treatment of acute and chronic toxoplasmosis in mouse models.
Materials and methods: CR-NE 1% was prepared using spontaneous emulsification by soybean as oil phase; a mixture of Tween 80 and Tween 85 as surfactant; ethanol as cosurfactant and distilled water. Particle size and zeta potential of NE were assessed using Nano-ZS90 dynamic light scattering. Stability testing of NE was assessed after storage for 2 months at room temperature. In vivo experiments were carried out using 50 BALB/c mice inoculated with virulent RH strain (type I) and 50 BALB/c mice inoculated with avirulent Tehran strain (type II) of Toxoplasma gondii and treated with CR-NE (1% w/v), CR suspension (CR-S, 1% w/v), and NE without CR (NE-no CR).
Results: The mean particle size and zeta potential of CR-NE included 215.66±16.8 nm and -29.46±2.65 mV, respectively, and were stable in particle size after a three freeze–thaw cycle. In acute phase experiment, the survival time of mice infected with RH strain of T. gondii and treated with CR-NE extended from 8 to 10 days postinoculation. The differences were statistically significant between the survival time of mice in CR-NE-treated group compared with negative control group (P<0.001). Furthermore, CR-NE significantly decreased the mean counts of peritoneum tachyzoites from 5,962.5±666 in negative control group to 627.5±73 in CR-NE-treated mice (P<0.001). Growth inhibition rates of tachyzoites in peritoneum of mice receiving CR-NE, CR-S, and NE-no CR included 90%, 21%, and 11%, respectively, compared with negative control group. In chronic phase experiment, the average number and size of tissue cysts significantly decreased to 17.2±15.6 and 31.5±6.26 µm, respectively, in mice inoculated with bradyzoites of T. gondii Tehran strain and treated with CR-NE compared with that in negative control group (P<0.001). Decrease of cyst numbers was verified by downregulation of BAG1 in treatment groups compared with negative control group with a minimum relative expression in CR-NE (1.12±0.28), CR-S (11.76±0.87), and NE-no CR (14.67±0.77), respectively, (P<0.001).
Conclusion: Results from the current study showed the potential of CR-S and CR-NE in treatment of acute and chronic toxoplasmosis in mouse models for the first time. However, CR-NE was more efficient than CR-S, and it seems that CR-NE has a potential formula for the treatment of acute and chronic toxoplasmosis, especially in those with latent bradyzoites in brain.
Keywords: curcumin nanoemulsion, curcumin suspension, Toxoplasma gondii RH strain, Toxoplasma gondii Tehran strain, soybean oil, blood–brain barrier, spontaneous emulsification
... Dahbi et al. reported the total absence of intra-cerebral cysts in mice who received thyme EO (20 μg), signifying that they blocked the appearance of the cysts. No abnormalities were observed in the control mice who received the EO of thyme [131]. Other studies on mice have shown that extracts of Nigella sativa oil in combination with pyrimethamine had a synergistic effect in the treatment of toxoplasmosis. ...
Toxoplasmosis is an infection caused by Toxoplasma gondii, a widespread zoonotic protozoan which poses a great threat to human health and economic well-being worldwide. It is usually acquired by ingestion of water contaminated with oocysts from the feces of infected cats or by the ingestion of raw or undercooked foodstuff containing tissue cysts. The oocyst can contaminate irrigation water and fresh edible produce. It is estimated that approximately one-third of the human population worldwide harbor this parasite. Infection with T. gondii is an important cause of diseases of the central nervous system and the eye in immunocompromised and immunocompetent individuals. The purpose of this study was to evaluate the efficacy and applicability of thermal (heating, cooking, freezing and low temperature), non-thermal (high pressure processing, ionizing irradiation and curing) and chemical and biochemical (disinfection, essential oils and biochemical methods such as enzymes, nanoparticles, antibiotics and immune response) treatments for the inactivation, inhabitation or to kill T. gondii in foodstuff intended for public consumption and under experimental conditions.
... Therefore, grape seed oil was selected to prepare nanoemulsion based on the hidrophilic-lipophilic balance of the oil. Benefits of essential oils to decrease tissue cyst number of T. gondii have already been documented (Dahbi et al., 2010). Various doses of atovaquone were used due to metabolic nature and sensitivity of T. gondii strains; from which, doses of 12.5-25 mg/kg were potential in most strains (Alves and Vitor, 2005;Tomavo and Boothroyd, 1995). ...
Treatment of toxoplasmosis is necessary in congenital form and immunocompromised patients. Atovaquone is a powerful suppressor of protozoan parasites with a broad-spectrum activity, but an extremely low water solubility and bioavailability. In this study, nanoemulsion of this drug was prepared with grape seed oil using spontaneous emulsification method to increase bioavailability and efficacy of atovaquone for treatment of toxoplasmosis. In vitro activity of atovaquone nanoemulsion against T. gondii, RH and Tehran strains, was assessed in HeLa cell culture. For in vivo assessment, BALB/c mice were infected with RH and Tehran strains and then treated with nanoemulsion of atovaquone, compared to that treated with free atovaquone. Concentration of atovaquone nanoemulsion showed in vitro anti-parasitic effects in both strains of T. gondii. Furthermore, oral administration of atovaquone nanoemulsion increased oral bioavailability, tissue distribution and mice survival time and reduced parasitemia and number and size of the brain cysts. Decrease of cyst numbers was verified by down regulation of BAG1 using real-time polymerase chain reaction (real-time PCR) assay. Effective therapeutic activity of atovaquone at a reduced dose is the major achievement of this study.
... Botanically, the essential oil of Thymus broussonetii Boiss (Arabic names are za'itra,za'atar el-hamir and ra'atar essouiri, whereas the English name is broussonet thyme) was administered to mice at 20 µg/ animal orally at the time of infection with T. gondii and for several days thereafter. This resulted in total absence of intracerebral cysts in treated mice (Dahbi et al., 2010). ...
... Antiprotozoal activity also was observed against Trypanosoma cruzi ( Santoro et al., 2007;Escobar et al., 2010a). Previous studies demonstrated anti- Toxoplasma activity versus the PRU strain of T. gondii using a phenol-terpenoid (carvacrol) compound ( Dahbi et al., 2010). However, there are no previously published reports regarding the effects of the other monoterpenes, such as thymol, against the ME49 strain of T. gondii in a murine model. ...
Toxoplasmosis is caused by Toxoplasma gondii an obligatory intracellular protozoan. Normally benign, T. gondii infections can cause devastating disease in immunosuppressed patients and through congenital infection of newborn babies. Few prophylactic and therapeutic drugs are available to treat these infections. The goal of the present study was to assess the anti-Toxoplasma effects in a congenital and non-congenital model of toxoplasmosis (using ME49 strain), besides assessing immunological changes, in vitro cytotoxicity and in vivo acute toxicity of commercial estragole and thymol. The congenital experimental model was used with intermediate stages of maternal infection. The serum levels of IgM, IgG, IL-10, IL-12 and IFN-γ were quantified from infected and treated C57Bl/6 mice. Estragole and thymol respectively exhibited low-to-moderate in vivo toxicity and cytotoxicity. Animals treated with estragol showed high IFN-γ and strong Th1 response. Both compounds were active against T. gondii ME49 strain. Furthermore, orally administered estragole in infected pregnant mice improved the weight of offspring compared to untreated controls. Subcutaneous administration of both compounds also increased the weight of mouse offspring born to infected mothers compared to untreated controls. Estragole and thymol display important anti-Toxoplasma activity. Further studies are needed to elucidate the mechanism of action of these compounds.
... To examine the use of essential oils (EOs) or their components as antiparasitic agents, Dahbi et al. extracted the EO of Thymus broussonetti Boiss (Lamiaceae), endemic to Morocco, by hydrodistillation and evaluated the effect of the EO on the production of cysts in the brains of OF1 mice infected orally with 200 cysts of the Prugniaud strain (PRU) of T. gondii [30]. No tissue cysts were found in the brains of animals treated with 20 g/mL of the EO, in contrast to the control animals, where 122 tissue cysts were observed on average. ...
Toxoplasma gondii is an intracellular parasite widely distributed in nature. Infection is asymptomatic in immunocompetent individuals; however, various clinical manifestations may occur in immunocompromised individuals. Although there are medications for the treatment of toxoplasmosis, such as pyrimethamine and sulfonamide, they cannot always be used due to adverse reactions or to therapeutic failures related to intolerance or malabsorption of drugs and to parasite drug resistance. In recent years, the search for new antimicrobial agents derived from plants has intensified because a quarter of synthetic drugs that are currently prescribed have been isolated from a plant source, demonstrating that natural products are important in the development of new drugs. A systematic literature search was conducted to evaluate the use of natural products as an alternative for the treatment of T. gondii infection. The search was conducted for the 2000-2014 period in Science Direct, Scopus, PubMed, EMBASE, and SciELO databases, using the following MeSH terms: anti-Toxoplasma activity, toxoplasma AND natural products, toxoplasma AND plant extracts. Ethnobotanical and experimental evidence (in vitro/in vivo) was found supporting the use of natural products as a source for the discovery of new therapies against T. gondii.
... Botanically, the essential oil of Thymus broussonetii Boiss (Arabic names are za'itra,za'atar el-hamir and ra'atar essouiri, whereas the English name is broussonet thyme) was administered to mice at 20 µg/ animal orally at the time of infection with T. gondii and for several days thereafter. This resulted in total absence of intracerebral cysts in treated mice (Dahbi et al., 2010). ...
The aims of the present study were to diagnose toxoplasmosis in pregnant ewes and women serologically and molecularly, treat naturally infected ewes, and diagnose congenital toxoplasmosis. Blood samples were taken from 30 and 60 pregnant ewes and women, respectively, and used for diagnosis of toxoplasmosis through Latex agglutination test (LAT). Seropositive samples were confirmed by PCR for detection of acute infection. Ten infected pregnant ewes were classified into two groups. The first group was treated with sulfadimidine 33.3%, 200 mg (0.6 ml) / kg.b.wt, and the other group was treated with normal saline. At titers ≥ 1:64, serological diagnosis indicated that 16 (53.33%) ewes and 29 (48.3%) women were seropositive and the seroprevalence increased in older ewe and younger women. Positive LAT samples were used for amplification of DNA and showed bands at 193 bp, analogues to that of the RH strain in 12 (40%) and 15 (25%) blood samples of ewes and women, respectively. All treated ewes with sulfadimidine 33.3% delivered healthy lambs with normal gestation period, whereas untreated ewes delivered 4 abortuses and 3 stillbirths. The tissue cysts were demonstrated microscopically in stained smears from tissues of dead fetuses. Local strain of T. gondii was isolated through intra peritoneal injection in mice from tissues of abortuses and stillbirths and maintained in the lab
... Recent studies have shown strong antibacterial, antifungal, antiviral, antiparasitic, spasmolytic and antioxidant activities of Thymus species. It has been considered that a part of these effects is due to the volatile constituents78910111213. Therefore, there is a considerable research interest in the chemical composition analysis of Thymus essential oils and their biological activities. ...
Thymus longicaulis C. Presl is a small aromatic perennial herb used as a traditional remedy for cold, flu and cough. Composition of the essential oil of T. longicaulis from Croatia and its in vitro antimicrobial activity against the most common respiratory pathogens were evaluated. The yield of essential oil obtained by hydrodistillation from aerial plant parts was 1.2%. According to the GC-MS analysis, a total of forty one compounds (99%) were identified. Thymol (46.3%), γ-terpinene (16.2%), thymyl methyl ether (11.4%), and p-cymene (9.4%) were the main components. Antimicrobial activity of the essential oil against six clinically isolated bacterial and yeast strains was determined using standard disc agar diffusion method and microdilution broth assay. The essential oil exhibited antimicrobial activity towards all tested respiratory pathogens. The most sensitive strains were Haemophilus influenzae and Streptococcus pneumoniae(MIC=0.78 mg/mL), while Staphylococcus aureus was the most resistant (MIC>25.00 mg/mL). Our results indicate that T. longicaulis essential oil could be effective against clinically relevant respiratory pathogens which have the ability to develop resistance to antimicrobial drugs.
Identification of drug target in protozoan T. gondii is an important step in the development of chemotherapeutic agents. Likewise, exploring phytochemical compounds effective against the parasite can lead to the development of new drug agent that can be useful for prophylaxis and treatment of toxoplasmosis. In this review, we searched for the relevant literature on the herbs that were tested against T. gondii either in vitro or in vivo, as well as different phytochemicals and their potential activities on T. gondii. Potential activities of major phytochemicals such as alkaloid, flavonoid, terpenoids and tannins on various target sites on T. gondii as well as other related parasites was discussed. It is believed that the phytochemicals from natural sources are potential drug candidates for the treatment of toxoplasmosis with little or no toxicity to human.
The objective of the current study was to systematically review papers discussing the efficacy of medicinal herbs
against Toxoplasma gondii. Data were systematically collected from published papers about the efficacy of herbs used against
T. gondii globally from 1988 to 2015, from PubMed, Google Scholar, ISI Web of Science, EBSCO, Science Direct, and Scopus.
Forty-nine papers were included in the current systematic review reporting the evaluation of medicinal plants against T. gondii
globally, both in vitro and in vivo. Sixty-one plants were evaluated. Most of the studies were carried out on Artemisia annua. The
second highest number of studies were carried out on Glycyrrhiza glabra extracts. RH and ME49 were the predominant parasite
strains used. Additionally, Swiss-Webster and BALB/c mice were the major animal models used. Alcoholic and aqueous extracts
were used more than other types of extracts. Natural compounds mentioned here may be developed as novel and more effective
therapeutic agents that improve the treatment of toxoplasmosis due to their lower side effects, higher availability, and better
cultural acceptance compared with those of the chemical drugs that are currently being used.
The objective of the current study was to systematically review papers discussing the efficacy of medicinal herbs against Toxoplasma gondii. Data were systematically collected from published papers about the efficacy of herbs used against T. gondii globally from 1988 to 2015, from PubMed, Google Scholar, ISI Web of Science, EBSCO, Science Direct, and Scopus. Forty-nine papers were included in the current systematic review reporting the evaluation of medicinal plants against T. gondii globally, both in vitro and in vivo. Sixty-one plants were evaluated. Most of the studies were carried out on Artemisia annua. The second highest number of studies were carried out on Glycyrrhiza glabra extracts. RH and ME49 were the predominant parasite strains used. Additionally, Swiss-Webster and BALB/c mice were the major animal models used. Alcoholic and aqueous extracts were used more than other types of extracts. Natural compounds mentioned here may be developed as novel and more effective therapeutic agents that improve the treatment of toxoplasmosis due to their lower side effects, higher availability, and better cultural acceptance compared with those of the chemical drugs that are currently being used.
Various species of Thymus L. are used for medical, flavouring and food preservation purposes. In the current study, the essential oils of inflorescences and leaves of four thyme species (T. broussonetii, T. pallidus, T. maroccanus and T. satureioides) from South-Western Morocco were analysed by simultaneous GC-FID and GC-MS. The main compound was carvacrol, except for T. pallidus (p-cymene) and T. satureioides leaves (borneol). Additionally, antibacterial activities were assessed by agar diffusion and agar dilution assay. Although the essential oils affected most of the Gram-negative test strains on a mediocre level, they exhibited strong activities against Gram-positive bacteria and several Gram-negative bacteria, thus supporting the traditional fields of application of thyme species.
Various species of Thymus L. are used for medical, flavouring and food preservation purposes. In the current study, the essential oils of inflorescences and leaves of four thyme species (T. broussonetii, T. pallidus, T. maroccanus and T. satureioides) from SouthWestern Morocco were analysed by simultaneous GC-FID and GC-MS. The main compound was carvacrol, except for T. pallidus (p-cymene) and T. satureioides leaves (borneol). Additionally, antibacterial activities were assessed by agar diffusion and agar dilution assay. Although the essential oils affected most of the Gram-negative test strains on a mediocre level, they exhibited strong activities against Gram-positive bacteria and several Gram-negative bacteria, thus supporting the traditional fields of application of thyme species. Zusammenfassung Viele Arten von Thymus L. werden als Heil-und Würzmittel und zur Haltbarmachung von Lebensmitteln verwendet. In der vorliegenden Studie wurden die ätherischen Öle der Blüten-stände und Blätter von vier Thymianarten (T. broussonetii, T. pallidus, T. maroccanus und T. satureioides) aus Südwest-marokko mittels simultaner GC-FID und GC-MS analysiert. Als Hauptbestandteil wurde Carvacrol identifiziert, außer für T. pal-lidus-(p-Cymen) und T. satureioides-Blätter (Borneol). Zusätzlich wurde die antibakterielle Wirkung mittels Agardif-fusions-und Agarverdünnungstest untersucht. Obwohl die ätherischen Öle auf den Großteil der Gram-negativen Stämme eher mäßig wirkten, zeigten sie eine starke Aktivität gegenüber Gram-positiven Bakterien sowie einigen Gram-negativen Bak-terien, wodurch die traditionellen Anwendungsgebiete der Thy-mianarten gerechtfertigt werden.
In the present study, the antifungal activity of selected essential oils obtained from plants used as spices was evaluated against both fluconazole-resistant and fluconazole-susceptible Candida spp. The Candida species studied were Candida albicans, Candida dubliniensis, Candida tropicalis, Candida glabrata, and Candida krusei. For comparison purposes, they were arranged in groups as C. albicans, C. dubliniensis, and Candida non-albicans. The essential oils were obtained from Cinnamomum zeylanicum Breyn, Lippia graveolens HBK, Ocimum basilicum L., Origanum vulgare L., Rosmarinus officinalis L., Salvia officinalis L., Thymus vulgaris L., and Zingiber officinale. The susceptibility tests were based on the M27-A2 methodology. The chemical composition of the essential oils was obtained by gas chromatography-mass spectroscopy and by retention indices. The results showed that cinnamon, Mexican oregano, oregano, thyme, and ginger essential oils have different levels of antifungal activity. Oregano and ginger essential oils were found to be the most and the least efficient, respectively. The main finding was that the susceptibilities of fluconazole-resistant C. albicans, C. dubliniensis, and Candida non-albicans to Mexican oregano, oregano, thyme, and ginger essential oils were higher than those of the fluconazole-susceptible yeasts (P<0.05). In contrast, fluconazole-resistant C. albicans and Candida non-albicans were less susceptible to cinnamon essential oil than their fluconazole-susceptible counterparts (P<0.05). A relationship between the yeasts' susceptibilities and the chemical composition of the essential oils studied was apparent when these 2 parameters were compared. Finally, basil, rosemary, and sage essential oils did not show antifungal activity against Candida isolates at the tested concentrations.
This study utilized infectivity bioassays in cats and mice to assess the efficacy of inactivation of Toxoplasma gondii cysts in experimentally infected sheep meat and tissues subjected to chilling, freezing, heating, microwave cooking, and gamma ray irradiation. Heating at 60 degrees C or 100 degrees C for 10 minutes, freezing at either -10 degrees C for 3 days or -20 degrees C for 2 days, or irradiation at doses of 75 or 100 krad was sufficient to kill tissue cysts. Meanwhile, neither cooking in a microwave nor chilling at 5 degrees C for 5 days was sufficient to kill tissue cysts.
The essential oils obtained from Thymus vulgaris L. harvested at four ontogenetic stages were evaluated for their biological activity and chemical composition. The thyme essential oils were tested for their inhibitory effects against nine strains of gram-negative bacteria and six strains of gram-positive bacteria. The bioimpedance method was chosen for studying the antibacterial activity of the essential oils and the parameter chosen for defining and quantifying the antibacterial activity of the essential oils was the detection time. The plate counting technique was used to study the inhibitory effect by direct contact. All the thyme essential oils examined had a significant bacteriostatic activity against the microorganisms tested. This activity was more marked against the gram-positive bacteria. The oil from thyme in full flower was the most effective at stopping the growth of the microbial species examined. The oils tested were also shown to have good antibacterial activity by direct contact, which appeared to be more marked against the gram-negative bacteria. Only a few of the species were capable of recovering at least 50% of their metabolic function after contact with the inhibitor, while most of the strains were shown to have been inactivated almost completely. Escherichia coli O157:H7 was the most sensitive species, given that after contact with even the lowest concentration of oil cells could not be recovered.
Carvacrol is a component of several essential oils and has been shown to exert antimicrobial activity. The structural requirements for the activity of carvacrol were determined by comparison to structurally related (nonessential oil) compounds. Removal of the aliphatic ring substituents of carvacrol slightly decreased the antimicrobial activity. The effect of the hydroxyl group of carvacrol on activity could not be determined by simply comparing it to p-cymene, because this compound is immiscible with water; therefore, 2-amino-p-cymene, the amino analogue of carvacrol, which has a similar hydrophobicity and structural characteristics, was used. 2-Amino-p-cymene had similar membrane disruption and bacterial killing characteristics as carvacrol showing that, contrary to previous reports, the hydroxyl group of carvacrol itself is not essential for the antimicrobial activity. However, the observed 3-fold lower activity for 2-amino-p-cymene as compared to carvacrol indicates special features in the antimicrobial mode of action of carvacrol due to the hydroxyl group.
In Morocco, Thymus broussonetii is widely used in folk medicine for the treatment of a variety of diseases including gastroenteric and bronchopulmonary disorders and to relieve dolorous process. The antinociceptive effect of the aqueous, butanolic and ethyl acetate extracts of this species was examined in rats and mice using chemical and thermal models. The results obtained showed that aqueous and butanolic extracts exerted an antinociceptive activity in the two phases of formalin (50-300 mg/kg), tail immersion and writhing tests. Whereas, the ethyl acetate extract reduced the nociceptive response only in the second phase of formalin (100-300 mg/kg) and writhing tests. The aqueous extract, which is the most effective, contains active analgesic principles acting both centrally and peripherally. Furthermore, this antinociceptive effect has been avoided by naloxone at a dose of 1mg/kg in the first phase of formalin and hot plate tests indicating that this extract acts partly through an opioid-mediated mechanism. The present results demonstrated that Thymus broussonetii contains active constituents which possess antinociceptive activity justifying its popular use to relieve some pains.
Enhanced or pumped pork products represent a significant proportion (40 to 50%) of the commercially available pork cuts available to consumers at the retail level. In a previous study, pork loins containing viable Toxoplasma gondii tissue cysts were pumped with solutions containing 2% sodium chloride or 1.4% or higher potassium or sodium lactate and stored at 4 degrees C for 7 days. This treatment prevented transmission of T. gondii to cats. In the present study, enhanced pork loins were stored for 0, 8, 16, 24, 32, or 40 h at 4 degrees C and then fed to T. gondii-seronegative cats to determine how quickly the loss of tissue cyst viability occurred. In a second experiment, pork loins collected from pigs experimentally infected with T. gondii were stored at temperatures found in retail meat cases and then fed to T. gondii-seronegative cats to determine the effect of typical meat case storage temperatures on tissue cyst viability. In both experiments, cat feces were examined for 14 days after the infected meat meal to assess oocyst shedding. The results indicate that solutions containing 2% sodium chloride or 1.4% potassium or sodium lactate are effective within 8 h of injection for killing T. gondii tissue cysts in pork loins and that storage at meat case temperatures at or below 0 degrees C (32 degrees F) for 7 days also killed T. gondii tissue cysts in pork loins.
The composition of the essential oil of Thymus pulegioides and its antifungal activity on Candida, Aspergillus and dermatophyte fungal strains were studied. Essential oil from the aerial parts of the plant was obtained by hydrodistillation and analysed by GC and GC-MS. The oil showed high contents of carvacrol and thymol. The MIC and minimal lethal concentration were used to evaluate the antifungal activity against Candida (seven clinical isolates and four ATCC type strains), Aspergillus [five clinical isolates, and two Colección Española de Cultivos Tipo (CECT) and two ATCC type strains] and five clinical dermatophyte strains. Antifungal activity was evaluated for the essential oil and for its main components. To clarify its mechanism of action on yeasts and filamentous fungi, flow-cytometric studies of cytoplasmic membrane integrity were performed, and the effect on the amount of ergosterol was investigated. Results showed that T. pulegioides essential oil exhibited a significant activity against clinically relevant fungi, mainly due to lesion formation in the cytoplasmic membrane and a considerable reduction of the ergosterol content. The present study indicates that T. pulegioides essential oil has considerable antifungal activity, deserving further investigation for clinical applications.
The anti-tumor effect of the Moroccan endemic thyme (Thymus broussonettii) essential oil (EOT) was investigated in vitro using the human ovarian adenocarcinoma IGR-OV1 parental cell line OV1/P and its chemoresistant counterparts OV1/adriamycin (OV1/ADR), OV1/vincristine (OV1/VCR), and OV1/cisplatin (OV1/CDDP). All of these cell lines elicited various degrees of sensitivity to the cytotoxic effect of EOT. The IC50 values (mean +/- SEM, v/v) were 0.40 +/- 0.02, 0.39 +/- 0.02, 0.94 +/- 0.05, and 0.65 +/- 0.03% for OV1/P, OV1/ADR, OV1/VCR, and OV1/CDDP, respectively. Using the DBA-2/P815 (H2d) mouse model, tumors were developed by subcutaneous grafting of tumor fragments of similar size obtained from P815 (murin mastocytoma cell line) injected in donor mouse. Interestingly, intra-tumoral injection of EOT significantly reduced solid tumor development. Indeed, by the 30th day of repeated EOT treatment, the tumor volumes of the animals were 2.00 +/- 0.27, 1.35 +/- 0.20, and 0.85 +/- 0.18 cm(3) after injection with 10, 30, or 50 microL per 72 h (six times), respectively, as opposed to 3.88 +/- 0.50 cm(3) for the control animals. This tumoricidal effect was associated with a marked decrease of mouse mortality. In fact, in these groups of mice, the recorded mortality by the 30th day of treatment was 30 +/- 4, 18 +/- 4, and 8 +/- 3%, respectively, while the control animals showed 75 +/- 10% of mortality. These data indicate that the EOT which contains carvacrol as the major component has an important in vitro cytotoxic activity against tumor cells resistant to chemotherapy as well as a significant antitumor effect in mice. However, our data do not distinguish between carvacrol and the other components of EOT as the active factor.
Medicinal plants are potential of antimicrobial compounds. The present study deals with the antibacterial activity of methanolic extract of Thymus daenensis. Aerial parts of the plant were collected from Alvand mountainside (Hamadan, Iran) in May 2005, air-dried and extracted by methanol. The dried extract was redissolved in methanol to make a 100 mg/ml solution and then filtered. Antibacterial activity of the extract was evaluated against various Gram-positive and Gram-negatives bacteria using disk diffusion technique. Blank paper disks were loaded with 40 microl of the methanol solution and then dried up. The impregnated disks were placed on Mueller-Hinton agar inoculated with bacterial suspension equal to 0.5 McFarland. The extract inhibited the growth Gram-positive bacteria, i.e., Staphylococcus aureus, Micrococcus luteus, Entrococcus faecalis, Streptococcus pyogenes, but it showed no activity against Gram-negative bacteria. The most significant effect was seen against S.aureus including MRSA, which are important nosocomial pathogens. MIC90 of the extract was determined against Gram-positive bacteria (3.12 mg/ml) and 11 MRSA strain (1.56 mg/ml).
Three chemotypes of Thymus longicaulis C. Presl subsp. chaoubardii (Reichenb. fil.) Jalas were collected from the same population growing in one m area. Their oils were examined by GC and GC/MS. Limonene (18.7%) and thymol (19–4%); geraniol (56.8%) and geranyl acetate (7.6%); linalool (63.1%) and α-terpinyl acetate (20.4%) were the predominant components in each of the three different chemotypes, respectively. The oils also exhibited moderate to strong antimicrobial activity against the six Gram (±) tested bacteria
The antifungal activity of commercial Thymus vulgaris oils on the development of mycelium of the the phytopathogenic fungi Fusariumsolani (Mart.) Sacc., Rhizoctoniasolani Kühn and Colletotrichumlindemuthianum (Sacc. et Magn.) Briosi et Cav. was studied. The gas chromatography data showed that the essential oils were rich in thymol (22–38%) and its biogenetic precursors γ-terpinene and p-cymene. The carvacrol content was relatively low in all the oils tested, ranging between 1% and 2%. The fungicidal activity of the oils was correlated with their thymol content.The principal chemical components of thyme oils were then tested using the same concentrations as in the thyme oils. Only the fungicidal activity of thymol was high and close to that of the oils with the same thymol content. The effects of the thyme oils and thymol on the hyphae cytomorphology of the studied fungi was an increased vacuolization of the cytoplasm and accumulation of lipid bodies, undulations of the plasmalemma, and alterations of the mitochondria and endoplasmic reticulum.
In the present study, the antifungal activity of selected essential oils obtained from plants used as spices was evaluated against both fluconazole-resistant and fluconazole-susceptible Candida spp. The Candida species studied were Candida albicans, Candida dubliniensis, Candida tropicalis, Candida glabrata, and Candida krusei. For comparison purposes, they were arranged in groups as C. albicans, C. dubliniensis, and Candida non-albicans. The essential oils were obtained from Cinnamomum zeylanicum Breyn, Lippia graveolens HBK, Ocimum basilicum L., Origanum vulgare L., Rosmarinus officinalis L., Salvia officinalis L., Thymus vulgaris L., and Zingiber officinale. The susceptibility tests were based on the M27-A2 methodology. The chemical composition of the essential oils was obtained by gas chromatography-mass spectroscopy and by retention indices. The results showed that cinnamon, Mexican oregano, oregano, thyme, and ginger essential oils have different levels of antifungal activity. Oregano and ginger essential oils were found to be the most and the least efficient, respectively. The main finding was that the susceptibilities of fluconazole-resistant C. albicans, C. dubliniensis, and Candida non-albicans to Mexican oregano, oregano, thyme, and ginger essential oils were higher than those of the fluconazole-susceptible yeasts (P<0.05). In contrast, fluconazole-resistant C. albicans and Candida non-albicans were less susceptible to cinnamon essential oil than their fluconazole-susceptible counterparts (P<0.05). A relationship between the yeasts' susceptibilities and the chemical composition of the essential oils studied was apparent when these 2 parameters were compared. Finally, basil, rosemary, and sage essential oils did not show antifungal activity against Candida isolates at the tested concentrations.
In the present study, four separate samples of Origanum onites L., three separate samples of Satureja thymbra L., Origanum vulgare L. ssp. hirtum (Link) Ietswaart, and Thymus cilicicus Boiss. & Bal. were collected from various regions of Mugla, Turkey. The essential oils of these plants were obtained by the hydrodistillation method. Antimicrobial activities of the essential oils against microorganisms, including multiple antibiotic-resistant bacteria, were investigated using the disc diffusion method. Different antibiotic discs were used for comparison to the inhibition zones. All the essential oils used in this study were very effective against Gram-positive and Gram-negative bacteria, which included multiple resistant strains, except Pseudomonas aeruginosa ATCC 27853 and Pseudomonas fluorescens MU 87. The essential oils of the O. onites, O. vulgare ssp. hirtum, and S. thymbra were especially very effective against the resistant strains such as Stenotrophomonas maltophilia MU 64, S. maltophilia MU 99, and Chryseomonas luteola MU 65. The maximum antimicrobial activity was observed with the essential oils of O. onites. The antimicrobial activities of the essential oils varied depending on the species, subspecies, or variety. In fact, the essential oils of some plants belonging to the same species that were collected from different locations showed different levels of antimicrobial activities.
The relationship between traditional and modern medicine should be seen as the evolvement of cosmopolitan medicine because the development of modern medicine is based on the achievements of many past civilizations. This article discusses the vital role that traditional medicine plays in Morocco--it is used by 80% of the population in concert with modern medicine. The decision to go to the fqih and the herbalist depends on such factors as cost, convenience, beliefs, personal prestige of the practitioner, and a person's own self-esteem. The majority of times, a traditional practitioner will be chosen over the medical system. The positive aspects of traditional medicine include such factors as similarities in background, culture and language between the practitioner and the patient; care is provided in patients' homes and the cure includes seeking a balance to any discords between health and disease. Health is perceived as a balance between the person's physical, mental, emotional, moral and social well-being. The Moroccan pharmacopeia is well-known to the people, and only has a few dangerous products. However, there are some negative aspects to traditional medicine. The concept of causality has not been refined and often misses the diagnosis; exact forms of measurement and administration are being lost and popularization or the ritualization of a medicine has made its use become too common (e.g. plants such as artemisia, thyme, marjoram, cumin). Public health administrators should study the advantages of integrating both health systems to achieve the best care possible by drawing on all existing resources in the country.
Flavonoids from Thymus vulgaris L. have been studied in vitro for spasmolytic activity on the smooth muscles of the guinea-pig ileum and trachea and of the rat vas deferens. The flavones and thyme extracts inhibit responses to agonists which stimulate specific receptors (acetylcholine, histamine, L-noradrenaline) as well as to agents whose actions are not mediated via specific receptors (BaCl2). Cumulative dose-response curves point to a non-competitive antagonism. Inhibition of Ca2+ contractions on K+ depolarized smooth muscles suggest inhibition of availability of Ca2+ for muscle contraction. pD'2 values were determined to compare the potency of these components to each other and to standard drugs. Flavones induce relaxation of the carbachol contracted tracheal strip without stimulation of the beta 2-receptors, which were blocked by propranolol.
A thyme extract, the preparation of which is described, produced a considerable stimulation of leucopoiesis and also an elevation of the thrombocyte count in the blood. In a comparative trial, the immunopotentiator 2-cyanoethylurea (BA1-4) has been found to be even more effective. The elevation of the leucocyte count is of importance not only to the various possibilities of immunostimulation, but also directly, in connexion with an elevation of the thrombocyte count, to the tumour-damaging process in the cancer multistage therapy, due to the promotion of the selective haemostasis set up on this occasion in the tumour tissue. The activation of immunological processes by intensive variants of the oxygen multistage therapy (O2-MT) increases in particular the efficiency of immunopotentiators so the latter should be used, in connexion with O2-MT, in any cancer therapy.
The antimicrobial activity of thymol, carvacrol, cinnamaldehyde and eugenol alone or combined was tested by micromethods on eight oral bacteria. The compounds showed an inhibitory activity on seven microorganisms and a synergistic effect was observed with certain combinations. The four compounds can be used alone or combined, as eugenol and thymol, eugenol and carvacrol, thymol and carvacrol, during the treatment of oral infectious diseases.
The survival of sporulated Toxoplasma gondii oocysts in water at -10 C to 70 C for various periods was investigated. Infectivity of T. gondii was tested by bioassay in mice. There was no marked loss of infectivity of oocysts stored at 10 C, 15 C, 20 C, and 25 C for 200 days, whereas there was a 100-fold loss of infectivity of oocysts stored at 30 C for 107 days. Oocysts stored at 35 C were infective for 32 days but not 62 days, at 40 C oocysts were infective for 9 days but not 28 days, at 45 C, oocysts were infective for 1 day but not 2 days, at 50 C oocysts were infective for 1 hr but not 2 hr. At 55 C and 60 C oocysts were rendered noninfective in 2 and 1 min, respectively. Oocysts remained infective up to 54 mo at 4 C and there was no loss of infectivity in oocysts stored for 106 days at -5 C and at -10 C and for 13 mo at 0 C.
Toxoplasma gondii oocysts are excreted nonsporulated in the feces of the cats into the environment. These oocysts must undergo sporulation to become infectious. Little is known about the factors that influence sporulation of T. gondii oocysts. The present study examined the survival of nonsporulated oocysts under refrigerated conditions over 11-week observation period. Microscopic examination of oocysts indicated that no visible development occurred under refrigerator conditions. The nonsporulated oocysts retained their ability to sporulate when placed at room temperature. The numbers of visually viable appearing oocysts decreased over time. Some oocysts in all samples were infectious for mice despite being refrigerated for up to an 11 weeks before undergoing sporulation. Results indicate that nonsporulated oocysts can survive in the environment for at least 3 months and retain their ability to become infectious when placed under appropriate conditions.
The topical anti-inflammatory activity of four extracts from Thymus broussonetii Boiss (Labiatae) leaves, a herbal drug used in Moroccan traditional medicine, has been studied using the croton oil ear test in mice. A bioassay-oriented fractionation revealed that the pharmacological activity is mainly in the chloroform extract. Fractionation and analysis of this extract allowed the identification of ursolic acid and oleanolic acid as the main anti-inflammatory principles. Some flavonoids (luteolin, eriodictyol, thymonin) and glycosides (luteolin-7-O-glucoside, luteolin-3'-O-glucuronide, eriodictyol-7-O-glucoside) were also isolated from the methanol extract.
Four extracts at increasing polarity were prepared from the leaves of Thymus satureioides Coss. (Labiatae) and assayed for the in vivo topical anti-inflammatory effect using the croton oil ear test in mice, and for in vitro both antioxidant (DPPH degrees test) and anti-bacterial (broth microdilution method) activities. The chloroform extract showed a topical anti-inflammatory activity (ID50=282 microg cm(-2)), only three times lower than that of the reference drug indomethacin (ID50=93 microg cm(-2)) and its active components were identified as ursolic and oleanolic acids. The methanol extract, showing a significant radical-scavenging effect (SC50=14.54 microg), was characterized by the isolation and identification of some flavonoids. On the contrary, the extracts did not show any anti-bacterial effect against four standard aerobial bacteria strains.
In the present work, we had tried to evaluate the immunotropic and behavioural effects of Thymus broussonetii Boiss. So, we tested the neurostimulant effects of four extracts. This preliminary study allowed to identify both the immunostimulant and the neurotropic antistress effects of the studied extracts. Among the four extracts, only the aqueous and ethyl acetate ones showed an apparent effect on the tested biological activities, whereas the butanolic extract and the essential oil did not show any interesting effect (data not shown). These results showed that the aqueous and ethyl extracts of this endemic species are of interest for two reasons: stimulation of the immunizing system and protection against the stress by a neurotropic activity. Thyme extracts increased in vivo the number of leucocyte categories studied including polynuclears, total lymphocytes, TCD4+, TCD8+ and NK cells. These data suggest that the intraperitoneal administration of Thymus broussonetii extract has a potent direct effect on leucocytes in vivo. The elevation of leucocyte and thrombocyte counts produced by thyme in the peripheral blood was already reported in the literature. These results could be of practical importance in the field of phytotherapy in the treatment of some cases of human immunodeficiency such as cancer, leukaemia and AIDS.
Tachyzoites, bradyzoites (in tissue cysts), and sporozoites (in oocysts) are the three infectious stages of Toxoplasma gondii. The prepatent period (time to shedding of oocysts after primary infection) varies with the stage of T. gondii ingested by the cat. The prepatent period (pp) after ingesting bradyzoites is short (3-10 days) while it is long (18 days or longer) after ingesting oocysts or tachyzoites. The conversion of bradyzoites to tachyzoites and tachyzoites to bradyzoites is biologically important in the life cycle of T. gondii and it has been proposed that the pp can be used to study stage conversion. In the present study, infectivity of oocysts and bradyzoites released from tissue cysts of a recent isolate of T. gondii, TgCkAr23, to cats and mice was compared. Ten-fold dilutions of oocysts or bradyzoites were administered orally to cats, and orally and subcutaneously to mice. Of the 29 cats each fed 1-10 million oocysts only one cat shed oocysts and the pp was 23 days; all cats remained asymptomatic. In contrast, all mice administered the same 10-fold dilutions of oocysts either orally or subcutaneously died of toxoplasmosis. The results confirm that infectivity of the oocysts to cats is lower than for mice and that oocysts are non-pathogenic for cats. Of the 41 cats each fed 1-1,000 free bradyzoites, 15 shed oocysts with a short pp of 4-9 days, and all remained asymptomatic. The infectivity of bradyzoites to mice by the oral route was approximately 100 times lower than that by the subcutaneous route. The results confirm the hypothesis that the pp in cats is stage and not dose dependent, and that transmission of T. gondii is most efficient when cats consume tissue cysts (carnivory) or when intermediate hosts consume oocysts (fecal-oral transmission).
The antimicrobial effect of thyme essential oil (EO) at 0.3%, 0.6%, or 0.9%, nisin at 500 or 1000IU/g, and their combination against Listeria monocytogenes was examined in both tryptic soy broth (TSB) and minced beef meat. Thyme EO at 0.3% possessed a weak antibacterial activity against the pathogen in TSB, whereas at 0.9% showed unacceptable organoleptic properties in minced meat. Thus, only the level of 0.6% of EO was further examined against the pathogen in minced meat. Treatment of minced beef meat with nisin at 500 or 1000IU/g showed antibacterial activity against L. monocytogenes, which was dependent on the concentration level of nisin and the strains used. Treatment of minced beef meat with EO at 0.6% showed stronger inhibitory activity against L. monocytogenes than treatment with nisin at 500 or 1000IU/g. All treatments showed stronger inhibitory activity against the pathogens at 10 degrees C than at 4 degrees C. The combined addition of EO at 0.6% and nisin at 500 or 1000IU/g showed a synergistic activity against the pathogen. Most efficient among treatments was the combination of EO at 0.6% with nisin at 1000IU/g, which decreased the population of L. monocytogenes below the official limit of the European Union recently set at 2logcfu/g, during storage at 4 degrees C.
Essential oils (EOs) and methanol extracts obtained from aerial parts of Thymus vulgaris and Pimpinella anisum seeds were evaluated for their single and combined antibacterial activities against nine Gram-positive and Gram-negative pathogenic bacteria: Staphylococcus aureus, Bacillus cereus, Escherichia coli, Proteus vulgaris, Proteus mirabilis, Salmonella typhi, Salmonella typhimurium, Klebsiella pneumoniae and Pseudomonas aeruginosa. The essential oils and methanol extracts revealed promising antibacterial activities against most pathogens using broth microdilution method. Maximum activity of Thymus vulgaris and Pimpinella anisum essential oils and methanol extracts (MIC 15.6 and 62.5mug/ml) were observed against Staphylococcus aureus, Bacillus cereus and Proteus vulgaris. Combinations of essential oils and methanol extracts showed an additive action against most tested pathogens especially Pseudomonas aeruginosa.
Chemical composition and antibacterial properties of Thymus longicaulis subsp. chaubardii oils: three chemotypes in the same population Spasmolytic activity of the flavonoids from Thymus vulgaris Structural requirements for the antimicrobial activity of carvacrol
- O E Verykokidou
- Roussis V Chinou
O, Verykokidou E, Roussis V, Chinou I (1998) Chemical composition and antibacterial properties of Thymus longicaulis subsp. chaubardii oils: three chemotypes in the same population. J Essent Oil Res 10:97–99 Van Den Broucke CO, Lemli JA (1983) Spasmolytic activity of the flavonoids from Thymus vulgaris. Pharm Weekbl Sci 25:9– 14 Veldhuizen EJ, Tjeerdsma-van Bokhoven JL, Zweijtzer C, Burt SA, Haagsman HP (2006) Structural requirements for the antimicrobial activity of carvacrol. J Agric Food Chem 8:1874–1879
Les plantes médiciles du Maroc
- A Sijelmassi
The effect of low temperature on the activity of Toxoplasma gondii cysts in sheep meat
- S A Michael
- Ahh El Rafaii
- T A Morsy
- SA Michael