In Vitro Anti-Leishmanial Activity of Glucosinolate Fraction from Alyssum linifolium Steph. ex Willd (Brassicaceae).

Objectives: The intracellular parasitic protozoan, Leishmania spp., causes several forms of diseases in humans. Cytotoxicity and emergence of new strains resistance to the current anti-leishmanial drugs have encouraged researchers to focus on new resources. Glucosinolates (GSL) are found mainly in the Brassicaceae family with potential cytotoxic and anti-parasitic properties. The present study reports in vitro antileishmanial activity of the GSL fraction from Alyssum linifolium seeds against Leishmania major. Materials and Methods: The GSL fraction was prepared by ion-exchange and reversed-phase chromatography. For the assessment of antileishmanial activity, the promastigotes and amastigotes of L. major were treated with different concentrations of the fraction (75-625 µg/mL). Results: The IC50 was 245 µg/mL for anti-promastigote effect of the GSL fraction and 250 µg/mL for its anti-amastigote effect that had a significant difference (p<0.05) with both glucantime and amphotericin B. The selectivity index of the GSL fraction (15.8), to glucantime and amphotericin B, was greater than 10, indicating the selective effect of this fraction against L. major amastigotes. Glucoiberverin was the major constituent of the GSL fraction characterized using nuclear magnetic resonance and electron ionization-mass spectrometry spectra. Based on gas chromatography-mass spectrometry data, iberverin and iberverin nitrile, the hydrolysis constituents from glucoiberverin, included 76.91% of the total seed volatiles. Conclusion: The results suggest that GSLs like glucoiberverin could be considered a new promising candidate for further studies on antileishmanial activity.

Anti-Toxoplasma gondii Activity of Curcumin: A Systematic Review of Pre-clinical Studies.

OBJECTIVE: This study has focused on anti-Toxoplasma gondii activity of curcumin. METHODS: In this systematic review, anti-parasitic activity of Curcuma longa on Toxoplasma gondii was assessed. Therefore, several databases, including PubMed, Scopus, Web of Science, Embase and Google Scholar, were searched from 2010 to 2020. RESULTS: Of the 2200 papers retrieved between 2010 and 2020, six articles were reliable and were scrutinized. In 2 in vitro studies, the most used strain was the RH strain of Toxoplasma gondii, whereas among 4 in vivo studies, RH strain was found in 2 (50%) studies, Me49 strain in 1(25%) study, and RH and Me49 strain in 1 (25%) study. In four in vivo studies, the most used animal model was BALB/c, and Swiss albino was found in 1 study (25%) and Albino rats in 1 study (25%). CONCLUSION: Curcumin and nanoparticles formulated with curcumin are new and useful agents for the treatment of parasitic diseases and reduction of drug resistance. The success of this therapeutic approach stems from the specific action of Curcuma longa against parasites and pathogens.

New strategies in the treatment of diseases caused by Acanthamoeba based on nanoparticles: a systematic review.

BACKGROUND: Acanthamoeba is one of the opportunistic parasites with a global prevalence. Currently, due to the side effects and the emergence of drug resistance to this parasite, much research has been performed on the use of nano-drugs to treat Acanthamoeba-caused diseases. Therefore, this systematic review study aims to evaluate new strategies for treating diseases caused by Acanthamoeba based on nanoparticles (NPs). METHODS: We designed a systematic review based on the articles published in English between 2000 and 2022. Our search strategy was based on syntax and specific tags for each database, including ScienceDirect, PubMed, Scopus, Ovid, and Cochrane. From the articles, those that had inclusion criteria were selected, and their data were extracted and analyzed. RESULTS: In this study, 26 studies were selected. Metallic nanoparticles were mostly used against the Acanthamoeba species (80.7%). 19.2% of the studies used polymeric nanoparticles, and 3.8% used emulsion nanoparticles. Most studies (96.1%) were performed in vitro, and only one study (3.8%) was carried out in vivo. Silver NPs were the most used metallic nanoparticles in the studies. The best effect of the anti-Acanthamoeba compound was observed for green synthesized nanoparticles based on stabilization by plant gums, loaded with citrus fruits flavonoids hesperidin (HDN) and naringin (NRG) with a 100% growth inhibition at a concentration of 50 µg/mL. CONCLUSION: This study showed that chlorhexidine and other plant metabolites loaded with silver and gold nanoparticles increase the anti-Acanthambae activity of these nanoparticles. However, green synthesized nanoparticles based on stabilization by plant gums, loaded with citrus fruits flavonoids hesperidin (HDN) and naringin (NRG), showed the best anti-Acanthambae effect. Nevertheless, further studies should be performed to determine their safety for human use.

First report of Lophomonas spp. in German cockroaches (Blattella germanica) trapped in hospitals, northern Iran.

Cockroaches are considered to be important reservoirs for protozoan and helminth parasites,particularly an emerging Lophomonas spp., protozoan parasite. German cockroaches, Blattella germanica (B. germanica), are domestic pest distributed worldwide. However, little information is available regarding the burden of Lophomonas spp. infestation among domestic cockroach population worldwide. The present study investigated parasitic agents, emphasizing Lophomonas spp. infestation among B. germanica in northern Iran. During spring and summer 2018, a total of 496 adults B. germanica (265 males and 231 females) were trapped by hand and glass traps from the teaching hospitals in Mazandaran, Sari, northern Iran. All cockroaches were identified using taxonomic keys. Each cockroaches was anesthetized with chloroform and the alimentary tract removed. Each section was smeared onto a glass slide. All the smears were examined under light microscopy to determine the presence of the parasites. Ten genera of parasites were identified from the trapped B. germanica; overall, 233 (47%) were found to be positive for helminths (n = 135) and protozoa (n = 98). The helminths included Aspiculuris tetraptera egg (67/233, 28.7%), Oxiuros sp. (39/233; 16.7%), Dentostomella translucida (17/233; 7.3%), and nematodes Ova (12/233; 5.2%). The protozoan parasites identified Gregarina sp. (61/233; 26.2%), Entamoeba sp. (19/233; 8.2%), Blastocystis sp. (5/233; 2.1%), Lophomonas (4/233; 1.7%), Nyctotherus sp. (4/233; 1.7%), unclassified flagella (5/233; 2.1%). Lophomonas spp. was detected within the gut of B. germanica, for the first time, in Iran. It has previously been identified in the gut of cockroaches a long time ago, however, this is the first attempt to detect Lophomonas in cockroaches housing in hospitals. Thereby, it possibly presenting pose a serious respiratory infection risk for patients and health care personnel.

Peganum harmala and Nigella sativa: anti-leishmanial activity against Leishmania major promastigotes and amastigotes: in vitro and ex vivo experiment.

Leishmaniosis is one of the most important vectors borne disease that is endemic in tropical and subtropical areas. There are many approved treatment for different types of leishmaniosis but all are with some adverse side effects that limited its uses. Here, we attempt to evaluate in vitro and ex vivo anti-leishmanial activities of Peganum harmala (P. harmala) and N. sativa (Nigella sativa) on promastigotes and amastigotes of L. major. The plants were extracted by maceration method and prepared in concentrations of 7.8, 3.9, 1.9, and 0.9 µg. L. major were cultured in RPMI-1640 medium alone and in J774 cell line separately. The extracts at different concentrations were assessed against promastigote (in vitro assay) and amastigotes (ex vivo assay) of L. major for 72 h at 22 and 37°C, respectively. In current work, N. sativa at highest concentration (7.8 µg/ml) showed 54.4 and 60% anti-leishmanial activity with IC50 of 5.3 and 3.278 µg/ml, respectively. Also, P. harmala at highest concentration (7.8 µg/ml) showed 68.9 and 58.6% antileishmanial activity with IC50 of 2.4 µg/ml for both of them, respectively. The SI value was 38.22 for N. sativa, 25.9 for P. harmala, 19.4 for Amphotericin B, and 16.33 for Glucantime. The results of our study indicated that N. sativa and P. harmala are effective against L. major promastigotes and amastigotes and could be consider as an alternative treatments for leishmaniosis. Therefore, it is recommended that further studies be performed to confirm the efficacy and evaluate the toxicity of the herbal extracts.

Seroprevalence of toxoplasmosis in Kermanshah City, west of Iran.

It is estimated that one-third of the world's population is infected with Toxoplasma gondii. The purpose of this study was to evaluate the latest status of toxoplasmosis seroprevalence in the general population and pregnant women in the west of Iran. This retrospective cross-sectional study was conducted in 2018. Accordingly, data associated with serodiagnosis of toxoplasmosis, age, sex, anti-toxoplasmosis IgG and IgM, and pregnancy status in women were collected from 6 health centers of Kermanshah City, the west of Iran, during 2016-2017. In total, 1228 people referred to the health centers in Kermanshah City. Of 1228 people, 359 (29.23%) individuals were seropositive for toxoplasmosis, of them 294 (81.89%) individuals were seropositive only for IgG, and 65 (18.11%) individuals were both IgG and IgM seropositive. The seropositivity in men was 29.3% (n = 63), in women was 29.2% (n = 296), and in pregnant women was 25.9% (n = 44). All individuals were examined using ELISA kit. This study showed that the prevalence of this disease in the west of Iran has been decreased in comparison with the previous studies. Therefore, regular epidemiological studies of in different regions seem to be necessary in order to conclude on the decrease or increase trend of this disease in an area.

Comparative evaluation of amniotic fluid as an alternative to fetal bovine serum in the maintenance of Leishmania major and Toxoplasma gondii.

This study aimed to evaluate the efficacy of amniotic fluid (AF) as an alternative to fetal bovine serum (FBS) in the maintenance of Leishmania major promastigotes and Toxoplasma gondii tachyzoites. AF was collected by an obstetrician using sterile syringes during a cesarean section. The culture medium was supplemented with 5 different concentrations of FBS or AF including 2, 5, 10, 20, and 30%. These concentrations were used to maintain both mentioned parasites. L. major was maintained at temperatures 4 and 24 °C and examined once a week for 4 weeks, while T. gondii was maintained at temperatures 4, 24, and 37 °C and examined at hours 24, 48, 72, and 96. For L. major, at both 4 and 24 °C, we observed no significant difference between FBS and AF on day 7. However, on days 14, 21, and 28, the difference between FBS and AF was significant at both temperatures. For T. gondii, no significant difference was observed between FBS and AF at hour 24 and all temperatures. However, this difference was significant at hours 48, 72, and 96 and all temperatures. According to our results, although FBS had a greater efficacy than AF in the growth of L. major and the survival of T. gondii, the number of promastigotes increased over time in AF-containing medium and the number of tachyzoites reduced slowly with a mild slop. Therefore, AF can be a potential alternative to FBS.

Apoptotic blebs from Leishmania major-infected macrophages as a new approach for cutaneous leishmaniasis vaccination.

We focused on apoptotic blebs from Leishmania major-infected macrophages as a vaccine for cutaneous leishmaniasis. Apoptosis was induced in L. major-infected J774A.1 cells in order to prepare apoptotic blebs. Test groups of BALB/c mice were immunized with these at doses of 1 × 106, 5 × 106 or 1 × 107 blebs. An immunization control group received Leishmania lysate antigens. The results showed that as the number of apoptotic bodies increased, the lymphocyte proliferation index increased, and this was proportional to IFN-γ level in the test groups. Additionally, the difference of IFN-γ, IL-4, IFN-γ/IL-4 ratio, or total IgG (p < 0.0001) in all groups was statistically significant compared to the negative control group. The highest IFN-γ (514.0 ± 40.92 pg/mL) and IFN-γ/IL-4 ratio (2.94 ± 0.22) were observed in the group that received 1 × 107 apoptotic blebs. The highest levels of IL-4 (244.6 ± 38.8 pg/mL) and total IgG (5626 ± 377 µg/mL) were observed in the immunization control group. Reflecting these data, no lesions were observed in any of the groups vaccinated with apoptotic blebs after 12 weeks. In summary, the use of apoptotic blebs from L. major-infected macrophages is protective against the challenge with L. major in this animal model.

Actin Gene-Based Molecular Typing of Trichomonas vaginalis Clinical Isolates from the North of Iran.

AIM: The aim of the current study is to evaluate the prevalence of trichomoniasis in men and women in the north of Iran and to find genotypes in the positive clinical specimens based on T. vaginalis actin gene. MATERIALS AND METHODS: Women's genital (n = 500) and men's urine (n = 1500) samples were collected from the participants referred to clinics in Mazandaran Province, northern Iran, during 2006-2018. In addition, 1500 Pap smear specimens, archived in the Bu Ali Hospital, Sari City, Mazandaran Province, northern Iran, were examined. The specimens were examined based on parasitological methods, nested polymerase chain reaction (PCR), PCR-restriction fragment length polymorphism, and phylogenetic analysis. RESULTS: Overall, 17 (0.48%) of 3500 specimens were positive by PCR. Total prevalence was 0.55% (n = 2000) for women, of which 500 (1.4%; n = 7) specimens were collected freshly, and 1500 (0.26%; n = 4) were Pap smears. Moreover, six (0.4%) out of 1500 men urine specimens were positive. Overall, genotypes G, E, and I were detected with the prevalence of seven (0.2%), seven (0.2%), and three (0.08%), respectively. There was no significant statistical difference among the prevalence of the detected genotypes (P > 0.05). CONCLUSION: As a whole, the prevalence of trichomoniasis was low in the studied area in the north of Iran and, most importantly, the genotypes of E, G, and I were distributed among men and women in the province.

Antileishmanial and antibacterial activities of the hydroalcoholic extract of Rhus coriaria L.

Leishmaniosis is one of the most serious public health concern with a worldwide distribution. Since the current treatments of leishmaniosis are toxic and expensive, frequent studies have been conducted to investigate the benefits of new resources such as medicinal plants for treatment of this infectious disease. Recent studies revealed the antiparasitic potential of Rhus coriaria. Here we investigated the potential antileishmanial and antibacterial activities of the hydroalcoholic extract of R. coriaria fruits. The fruits were extracted using 80% methanol by maceration method. The concentrations of 312, 156, 78, and 37 µg/ml of the extract were added separately to the wells containing Leishmania major (L. major) promastigotes and amastigotes. Amphotericin B was considered as positive control. Finally, the death rate was determined for the extract-treated parasites as compared to the non-treated parasite. The antibacterial activity was evaluated by measurement of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract against a set of Gram-positive and Gram-negative bacteria. The extract significantly inhibited the growth of both promastigotes (60,7%) and amastigotes (59%) at the concentration of 312 µg/ml with the IC50 values of 147 µg/ml and 233 µg/ml, respectively. The extract showed bactericidal effects against Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Acinetobacter baumannii. Totally, Grampositive bacteria were more susceptible to the extract. Our findings show that the hydroalcoholic extract of R. coriaria fruits are rich in tannins and can be considered for further in vivo studies on the antileishmanial and antibacterial activities especially on dermal lesions caused by L. major.

Tropolone alkaloids from Colchicum kurdicum (Bornm.) Stef. (Colchicaceae) as the potent novel antileishmanial compounds; purification, structure elucidation, antileishmanial activities and molecular docking studies.

Natural compounds played an important role for prevention and treatment of the disease as well as are the important compounds for the design of the new bioactive compounds. In this study, eight tropolone alkaloids were isolated from Colchicum kurdicum including colchicoside, 2-demethyl colchicine, 3-demethyl colchicine, demecolcine, colchifoline, N-deacetyl-N-formyl colchicine, colchicine and cornigerine by column and preparative thin layer chromatography. The chemical structures were identified by 1H NMR and 13C NMR spectroscopy. Moreover, the antileishmanial activity on Leishmania major, anti-inflammatory activity, iron chelating activity and toxicity studies including hemolytic activity, brine shrimp toxicity, cytotoxicity and acute toxicity and docking study of all isolated bioactive compounds were evaluated. As result, colchicoside and colchicine had potent leishmanicidal effects and N-deacetyl-N-formyl colchicine and cornigerine had the highest anti-inflammatory effects. All compounds had the significant iron chelating activity. According to toxicity studies, isolated compounds showed the low hemolytic activity and cytotoxicity, high LC50, LC90 and LD50. In the molecular docking study, colchicoside had the high dockscore. According to the study, with future studies all isolated compounds could be used for design the novel antileishmanial drugs.

Status of fasciolosis among domestic ruminants in Iran based on abattoir data: a systematic review and meta-analysis

Fasciolosis is considered as an ongoing neglected zoonotic disease in tropical regions of the world, relating to notable financial and public health issues. The current systematic review and meta-analysis was planned to determine the status of fasciolosis among domestic ruminants, including cattle, buffalo, goat, and sheep at the industrial slaughterhouses in Iran, between the years 2000 and 2016. Eight databases, four English and four Persian, were searched. Our findings demonstrated that 2.6% of all inspected livers of slaughtered ruminants were infected with Fasciola spp. during 2000­2016. The mean prevalence of fasciolosis for cattle, buffalo, goat, and sheep was 21%, 4.2%, 2%, and 2.4%, respectively. Additionally, most studies were performed on cattle 25 (39.6%), sheep 24 (38 %), and goat 12 (19%), respectively, and just 2 (3.1%) studies were performed on buffalo. The prevalence of animal fasciolosis has significantly decreased among domestic ruminants in Iran except for cattle. In addition, the prevalence of this disease in Northern and Western regions of the country has remained at hypo-endemic level. The results present updated gathered information on the epidemiology of fasciolosis in domestic ruminants in Iran, and will expand the screening strategies to improve health and reduce economic impacts among farm animals.

Human urogenital myiasis: A systematic review of reported cases from 1975 to 2017.

The public health importance of myiasis [infestation with dipterous (fly) larvae] remains unknown. This disease is spread worldwide in animals and humans, but baseline data on its prevalence are limited. In particular, knowledge on human urogenital myiasis (UGM) is scattered. As such, a systematic search was undertaken of five English and five Persian databases for publications describing UGM cases in English or Persian published between 1975 and 2017. In total, 45 papers reporting 59 UGM cases from various regions of the world are included in this review. All included papers were from the English databases. The age of patients ranged from 5 to 89 years, and the mean age was 40.6 years. Thirty-six of the patients were female and 19 were male. The highest number of cases (n = 12) was reported from Brazil. The most common genera causing UGM were Psychoda spp. (23.7%) and Cochliomyia spp. (11.8%). The vagina was the most commonly reported anatomical location of UGM for women, and the urogenital tract was the most commonly reported location for men. Thirteen cases were reported from rural areas and eight cases from urban areas; the location of other cases was not specified. The incidence of UGM is likely to be substantially underestimated when evaluated based on published case reports. Epidemiological studies, such as questionnaires to medical doctors, could help to gather the necessary baseline data on the occurrence of UGM.

Potent in vitro antileishmanial activity of a nanoformulation of cisplatin with carbon nanotubes against Leishmania major.

OBJECTIVES: The aim of this study was to evaluate the cytotoxicity and antileishmanial activity of cisplatin-bonded carbon nanotubes both against promastigotes and amastigotes of Leishmania major in vitro. METHODS: Cisplatin-bonded single-walled carbon nanotubes (CP-SWCNT) and cisplatin-bonded multi-walled carbon nanotubes (CP-MWCNT) were considered as test compounds. In addition, SWCNT, MWCNT, free cisplatin and meglumine antimoniate (Glucantime®) were considered as controls. The effect of each compound was evaluated both on promastigote and amastigote stages of L. major and the results were compared. RESULTS: There was a statistically significant difference between the half-maximal inhibitory concentration (IC50) of CP-SWCNT and each of the controls, including SWCNT, cisplatin and Glucantime® (P<0.05). In addition, IC50 values of CP-MWCNT and each of the controls, including MWCNT, cisplatin and Glucantime®, were significantly different both for promastigotes and amastigotes (P<0.05). However, the selectivity index (SI) of CP-SWCNT was <10 (5.23), indicating that this compound is not completely safe. Moreover, the SI values of CP-MWCNT (12.54) and Glucantime® (16.28) were >10, indicating the selective effect of these two compounds on the parasite. Moreover, the IC50 of CP-MWCNT (0.11±0.09µM) for amastigotes was 41-fold lower than that of Glucantime® (4.52±1.31µM), suggesting that a lower dose of CP-MWCNT in comparison with Glucantime® is required to kill 50% of amastigotes. CONCLUSIONS: According to the potent in vitro antileishmanial activity of CP-MWCNT at low concentration against L. major, we suggest that they are evaluated in an animal model.

Evaluating of Wistar rat and BALB/c mouse as animal models for congenital, cerebral and ocular toxoplasmosis.

This study was conducted to evaluate the potential of cyst production by Toxoplasma (T.) gondii, RH strain, in Wistar rat and BALB/c mouse and the purpose of this study was to introduce an animal model suitable for congenital, cerebral, and ocular toxoplasmosis. The mice and rats, considered as cerebral and ocular toxoplasmosis models, were intraperitoneally infected by different number of the parasite and their eyes and brain were evaluated for the presence of T. gondii cyst using the microscopic examination and the bioassay method. Moreover, the pregnant mice and rats, considered as congenital toxoplasmosis models, were intraperitoneally infected by different number of the parasite and their infants were examined by the method mentioned above. The best result for the cerebral toxoplasmosis model was observed in the rats infected with the 107 parasites, so that all infants (100%) were infected with the parasite when examined using the bioassay method. Furthermore, the best result was observed for the congenital cerebral toxoplasmosis model with 100% infection rate in the infants born to mothers infected with the 107 parasites. Overall, just few the ocular samples were positive using bioassay method. The best result in the current study was for the congenital cerebral toxoplasmosis model where the pregnant rats were infected with the 107 parasites and all infants were infected (100%). Therefore, these infants can be used as a congenital cerebral toxoplasmosis model when they are in the fetal stage, and can be used as a cerebral toxoplasmosis model one month after birth.

Vaccination against Toxoplasma gondii using rhoptry antigens: a systematic review.

Toxoplasma gondii is an obligate intracellular parasitic protozoan that infects a wide variety of vertebrates as intermediate hosts. The aim of the current systematic review study is to clarify the latest status of studies in the literature regarding rhoptry-associated recombinant proteins or rhoptry-associated recombinant DNAs as potential vaccines against toxoplasmosis. The search was performed systematically in 8 databases, four in English and four in Persian, up to February 2017. Overall, ROP2 was the most commonly used ROPs in DNA vaccines (27.27%) and protein vaccines (6.81%). Furthermore, regarding the type of adjuvants, route and dose of vaccination, animal models, challenge methods, and measurement of immune responses has been discussed in the text. It is hoped that this article help researchers to conduct more effective studies in the field of immunization against T. gondii.

Investigating in vitro anti-leishmanial effects of silibinin and silymarin on Leishmania major

Cutaneous leishmaniosis is an important zoonotic disease caused by various Leishmania species. The aim of this study was to investigate the effects of silibinin and silymarin on the in vitro growth and proliferation of promastigotes and amastigotes of Leishmania major compared to glucantime-treated parasites. The promastigotes and amastigotes of this parasite were treated with the two drugs, silibinin and silymarin, in several concentrations (25­100 µM). The highest effect on promastigotes was for silymarin in concentration of 100 µM with 90% and 91% death rate at hours 48 and 72, respectively. Regarding amastigotes, the highest effect at 48 hours was for silibinin in concentration of 100 µM with 35% death rate. However, at 72 hours, silymarin showed the highest effect with 63% death rate in concentration of 100 µM. The highest observed maximal 50% lethal concentration (LC50) for promastigotes was for silymarin with 19.34 µM at 48 hours and 18.22 µM at 72 hours. Likewise, maximal LC50 for amastigotes was for silymarin with 191 µM at 48 hours and 24.27 µM at 72 hours. Our findings demonstrated that both medications have suitable effects like Glucantime® on the parasite in vitro. Therefore, clinical assessment of the anti-leishmanial activity of silibinin and silymarin for treating the dermal lesions caused by L. major is recommended.

In vitro anti-leishmanial activity of Satureja hortensis and Artemisia dracunculus extracts on Leishmania major promastigotes.

Cutaneous leishmaniasis is one of the important skin diseases with diverse clinical manifestations. With an incidence of 0.7-1.3 million cases annually, this disease is often reported from six countries, including Iran. Accordingly, the purpose of this study was to evaluate the anti-leishmanial effect of the three plant hydroalcoholic extracts including fleawort (Plantago psyllium L.), savory (Satureja hortensis L.) and tarragon (Artemisia dracunculus L.) on Leishmania major promastigotes. The hydroalcoholic extract from each plant was extracted and its anti-leishmanial effect was evaluated in different concentrations (100-1000 µg/ml) and at various hours (24, 48 and 72 h). Savory herb inhibitory concentration 50 % (IC50) at 24, 48 and 72 h was 790.81, 398.11 and 298.42 µg/ml, respectively. In addition, tarragon herb IC50 at 24, 48 and 72 h was 962.03, 688.36 and 585.51 µg/ml, respectively. Moreover, the fleawort extract was showed the lowest effect, considering that its effect at the concentration of 1000 µg/ml was 48 % after 72 h (P > 0.05). Furthermore, the statistical analysis showed a significant difference for interaction between concentration and time regarding the tarragon and savory extracts with a P value of lower than 0.05. According to the results, the anti-leishmanial effect of the tarragon and savory extracts may make it possible to use them in the treatment of cutaneous leishmaniasis as a complementary or alternative therapy; however, further studies are necessary and should be evaluated in cell culture and in vivo conditions to confirm it.

Manipulation of IL-10 gene expression by Toxoplasma gondii and its products.

BACKGROUND: This study was designed to evaluate whether or not T. gondii and its derivatives can change the gene expression level of IL-10 in murine leukocytes in vivo. METHODS: Fifty BALB/c mice were divided into 5 groups, four of which received the excretory/secretory product (ESP) from cell culture medium, the ESP from cell free medium, the Toxoplasma lysate product (TLP) and the active tachyzoites, respectively. The fifth group was considered as control and received PBS. The peritoneal leukocytes from the mice were collected. Their total RNA were extracted and converted to cDNA and the gene expression levels of IL-10 in the samples were evaluated by quantitative real time-PCR using the REST-2009 software. RESULTS: The findings showed a decrease in the expression level of IL-10 in the TLP group (p=0.004). Moreover, the IL-10 gene expression level was upregulated in the group of the ESP from cell culture medium (p=0.04) and the active tachyzoite group (p=0.04). The expression of IL-10 gene in the group of ESP from cell-free medium was not significant compared to the control one (p=0.45). CONCLUSION: T. gondii and its derivatives are able to increase (the active T. gondii tachyzoite and the ESP from cell culture medium) and decrease (the TLP) the gene expression level of IL-10 in a murine model. The question remains to be examined in further study about which molecules are involved in this process.

Comparison of Eight Cell-Free Media for Maintenance of Toxoplasma gondii Tachyzoites.

BACKGROUND: Toxoplasmosis is considered as one of the most common infectious diseases caused by the protozoan parasite Toxoplasma gondii. Tachyzoite is the main form of Toxoplasma and continuously is maintained in cell culture or injected into the mice peritoneal cavity. This study was designed to evaluate the survival rate of RH strain of T. gondii tachyzoites in different cell free, nutrient and biological media at different temperatures. METHODS: This experimental study was performed at the Toxoplasmosis Research Center, Mazandaran University of Medical Sciences, Sari, Iran, in 2010. One ml of each solution including hypotonic saline (0.3%), normal saline (0.85%), RPMI-1640 (RPMI), RPMI with 10% fetal bovine serum (FBS), RPMI with 20% FBS, ovine hydatid cyst fluid, pasteurized milk of cow, and phosphate buffered saline (PBS) along with 4×10(4) T. gondii tachyzoites were added to plate wells and incubated in 4 °C, 22 °C, 37 °C, and 37 °C under 5% CO2. The survival rate and viability assessment of parasites were performed daily and the results were analyzed using Univariate tests. RESULT: Tachyzoites survival rate in PBS (4 °C) and normal saline (4 °C) were considerably high, compared to other solutions in different conditions (P<0.001). The best temperature for Toxoplasma maintenance was 4 °C (P<0.001). CONCLUSION: This study introduces two available and economical solutions, PBS (4 °C) and normal saline (4 °C) media, for maintenance of Toxoplasma tachyzoites as appropriate choice media for a noticeable period of time (11 days) in vitro.