Echinococcus granulosus sheep strain (G1) as the predominant genotype in definitive host (dogs) isolates in northeastern Iran.

Echinococcus granulosus sensu lato (E. granulosus s.l.) is a zoonotic parasite, causing cystic echinococcosis in humans. In the present study, prevalence and genotypes of E. granulosus s.l. was assessed in stools collected from 244 dogs including 138 stray and 106 domestic animals using high resolution melting curve (HRM) method. Initially, to detect taeniid eggs in feces, all samples were examined using the formalin-ether techniques. Genomic DNA was extracted from the positive samples and E. granulosus s.l. was differentiated from other Taeniidae parasites using SSU-rDNA gene and E. granulosus s.l. was analyzed for genotyping using HRM based on the cox1 gene. In total, 12.7% (31/244) of the samples were positive for Taeniidae eggs. In addition, among the positive samples, 77.4% (24/31) were positive for E. granulosus s.l.. In details, 11.3% (12/106) of the domestic dogs and 8.7% (12/138) of the stray dogs were positive for E. granulosus s.l.. The results of HRM analysis showed that all E. granulosus s.l. isolates were G1 strain. Findings of the present study indicated a considerable prevalence of E. granulosus G1 among dogs in the northeast of Iran and imply a serious risk of transmitting to humans and livestock.

A Systematic Review on Curcumin and Anti-Plasmodium berghei Effects.

BACKGROUND: Turmeric (Curcuma longa L.) is a popular spice containing curcumin that is responsible for its therapeutic effects. Curcumin with anti-inflammatory, antioxidant, anti-cancer, and antimicrobial activities has led to a lot of research focusing on it over the years. This systematic review aimed to evaluate research on the anti-Plasmodium berghei activity of curcumin and its derivatives. METHODS: Our study was performed according to PRISMA guidelines and was recorded in the database of a systematic review and preclinical meta-analysis of CAMARADESNC3Rs (SyRF). The search was performed in five databases, namely Scopus, PubMed, Web of Science, EMBASE, and Google Scholar, from 2010 to 2020. The following keywords were searched: "Plasmodium berghei", "Medicinal Plants", "Curcumin", "Concentration", Animals kind", "Treatment Durations", "Routes of Administration" and "in vivo". RESULTS: Of the 3,500 papers initially obtained, 14 articles were reliable and were thus scrutinized. Animal models were included in all studies. The most commonly used animal strain was Albino (43%), followed by C57BL/6 (22%). The other studies used various murine strains, including BALB/c (14%) and ICR (7%). Two (14%) studies did not mention the strain of animal model used. Curcumin alone or in combination with other compounds depending on the dose used, route of administration, and animal model showed a moderate to strong anti-Plasmodium berghei effect. CONCLUSION: According to the studies, curcumin has anti-malarial effects on Plasmodium berghei, and, however, its effect on human Plasmodium is unclear. Due to the side effects and drug resistance of current drugs in the treatment of human malaria, the use of new compounds with few or no side effects, such as curcumin, is recommended as an alternative or complementary treatment.

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.

Morphological and Molecular Identification of Emerged Lophomonas blattarum Infection in Mazandaran Province, Northern Iran: First Registry-Based Study.

BACKGROUND: In the last decade, several cases of bronchopulmonary lophomoniasis (BPL) have been recorded. Little information is available about epidemiological aspects on Lophomonas infection among BPL patients. The present study was aimed to investigate the prevalence of Lophomonas spp. infection in patients who were referred to the Iranian National Registry Center for Lophomoniasis (INRCL), using morphological and molecular tests. SUBJECTS AND METHODS: We examined patients enrolled in the INRCL from 2017 to 2019 at the Mazandaran University of Medical Sciences, northern Iran. All bronchoalveolar lavage fluid (BALF) and two nasal discharges of the patients were examined by both microscopic and small-subunit ribosomal RNA (SSU rRNA) PCR methods. To confirm the species of Lophomonas, two positive samples were sequenced. RESULTS: In this study, 321 specimens (including 319 BALF and 2 nasal discharges) were microscopically examined. Lophomonas spp. was found in 45(14%) (n = 44 BAL; n = 1 nasal discharge). The mean age of infected patients was 54.9 ± 17.1 years. The following morphological characteristics were observed in both fresh and Papanicolaou-stained smears to identify Lophomonas spp. All microscopically positive specimens were confirmed with genus-specific PCR technique. The obtained sequences were deposited in Gen Bank under the accession numbers (MN243135-36). The BLAST analysis of our two sequences with the only available sequence in the Gen Bank of the Thailand strain of L. blattarum, showed identity of 99-100% and 98.51%, respectively. CONCLUSION: To the best of our knowledge, this is the first registry-based study regarding lophomoniasis worldwide. According to our study, the conventional PCR test is an available and reliable tool for confirming the Lophomonas parasite in clinical samples. Moreover, the results confirmed that L. blattarum is circulating at least in our region.

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.

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.

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.

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.

Electrophoretic Patterns of Toxoplasma gondii Excreted/Secreted Antigens and Their Role in Induction of the Humoral Immune Response.

BACKGROUND: Toxoplasma gondii is an obligatory intracellular protozoan parasite on which studies are pending regarding production of vaccine. To date, the production of human vaccine has not been successful where approximately one third of the world's population is thought to be infected with T. gondii. OBJECTIVES: The present study was designed to compare the electrophoretic patterns of T. gondii excreted/secreted antigens (ESAs) and determine their role in the stimulation of the humoral immune response. MATERIALS AND METHODS: T. gondii ESAs were prepared from cell cultures (albino rat fibroblast) and cell-free mediums (RPMI-1640). Next, the SDS-PAGE technique was used for comparing molecular weights of the antigens. Forty C57BL/6 mice were divided randomly into four groups (n = 10). Immunization was performed subcutaneously at an interval of 2 weeks in two groups by injecting 100 µg of each of the above-mentioned antigens. Two groups, as negative control, also received fibroblast lysate proteins or adjuvant separately. All of the groups were then challenged with the T. gondii RH strain. Serum samples were collected from all mice and measured by immunoblotting technique for detection of immunogenic antigens. RESULTS: The electrophoretic mobility of the prepared antigens/proteins from cell culture, cell-free media, Fibroblast Lysate Proteins and Toxoplasma Lysate Antigens (TLA) showed 13, 12, 8 and 8 bands, respectively. The case groups, in challenge with T. gondii (RH strain), showed more survival prolongation than the control groups. Furthermore, the survival period was identical for both case groups with a tendency for slightly higher survival of mice receiving ESA from cell-free medium. Analysis of sera by immunoblotting also revealed one band of 65 KDa in sera from both case groups. CONCLUSIONS: We suggest that this band be extracted and its amino acids sequence determined to produce Synthetic Polypeptide for immunization studies.