Isolation of Acanthamoeba spp. from different water sources in Isfahan, central Iran, 2014.

Acanthamoeba spp. are free-living amoeba found in a wide variety of natural habitats. The high percentage of the presence of Acanthamoeba in different environmental sources represents a sanitary risk for public health, especially immunocompromised patients and contact lens wearers. Acanthamoeba can cause granulomatous amoebic encephalitis, otitis, lung lesions, and skin infections in individuals with immune deficiencies. In the present study, the status of contamination of water sources in Isfahan, central Iran is analyzed through parasitological method. Totally 93 samples were utilized consisting of 59 samples of tap water and 34 samples of environmental water collected from Isfahan in May and June 2014. After filtering, cultivation was done in non-nutrient agar medium, and then the cultured media were kept at 25-30 °C. The samples were analyzed based on the morphological criteria. Acanthamoeba spp. were found in 25 (73.53 %) out of 34 environmental water samples and 17 (28.8 %) out of 59 tap water. Generally, Acanthamoeba spp. were found in 42 (45.16 %) of the samples. The results of the present study showed that the water contamination with Acanthamoeba spp. in different regions of Isfahan can be a potential infection source for at high risk people. It could be suggested that public education and precaution are quiet necessary.

Antileishmanial activity of prenylated coumarins isolated from Ferulago angulata and Prangos asperula.

Leishmaniasis has a wide spectrum of signs and symptoms due to infection to numbers of Leishmania species and makes enormous mortality and morbidity. There are clues of antileishmanial effects of prenylated coumarins. Apiaceae family is one of the most important sources of coumarins. Air-dried aerial parts of Ferulago angulata and fruits of Prangos asperula were extracted with n-hexane, using a soxhlet apparatus. The solvents were evaporated under reduced pressure. Column chromatography and crystallization process resulted to isolation of three prenylated coumarins. (1)H-nuclear magnetic resonance, electron ionization Mass and Infrared spectra were used for elucidation of isolated compounds. Leishmanicidal activity of isolated coumarins was assessed on Leishmania major strain (MRHO/IR/75/ER) for the first time. Suberosin epoxide and suberosin were isolated from aerial parts of F. angulata and osthol was extracted from grounded fruits of P. asperula. Osthol showed a significant antileishmanial effect on promastigotes in early hours of exposure with IC50 of 14.40 µg/mL but suberosin epoxide showed only a weak antileishmanial activity. IC50 of osthol and suberosin epoxide after 48 h were 10.79 and 54.0 µg/mL, respectively. Suberosin showed no remarkable effect in these concentrations. This is the first report on the pharmacological activity of suberosin epoxide. Substantial difference between efficacies of two isomers, osthol and suberosin remarks the importance of prenyl substituent location on C-8.

A simple, efficient and inexpensive method for malaria parasites' DNA catching from fixed Giemsa-stained blood slides.

As parasitological or microscopic method is the gold standard and the best method for diagnosis of malaria, so fixed Geimsa-stained blood slides in the form of thick and thin blood smears are the most important data collections of malaria, especially historical slides. The parasites are dead but their DNA is valuable for many molecular biologic researches. A simple and efficient method for catching and extraction malaria parasites' DNA with a desired yield from dried and stained blood on slides is the first and major step. Introduction of an applicable, efficient and inexpensive DNA catching method and assessment of its performance in following molecular applications  was the main objective of present study.

Anti-Leishmania Activity of Osthole.

BACKGROUND: Treatment of cutaneous leishmaniasis (CL) is occasionally highly resistant to pentavalent antimonials, the gold standard in pharmacotherapy of CL. Since there is no effective vaccine, the discovery of natural antileishmanial products as complementary therapeutic agents could be used to improve the current regimens. OBJECTIVE: In this study in vitro and in vivo antileishmanial activities of osthole, a natural coumarin known to possess antibacterial and parasiticidal activities are evaluated. MATERIALS AND METHODS: Leishmania major infected J774.A1 macrophages were treated with increasing concentrations of osthole. CL lesions of BALB/c mice were treated topically with 0.2% osthole. RESULTS: Osthole exhibited dose-dependent leishmanicidal activity against intracellular amastigotes with IC50 value of 14.95 µg/ml. Treatment of CL lesions in BALB/c mice with osthole significantly declined lesion progression compared to untreated mice (P < 0.05), however did not result in recovery. CONCLUSION: Osthole demonstrated remarkable leishmanicidal activity in vitro. Higher concentrations of osthole may demonstrate the therapeutic property in vivo. SUMMARY: In vitro and in vivo antileishmanial activities of osthole, a pernylated coumarin extracted from Prangos asperula Boiss., are studied against Leishmania major.

Seroprevalence of toxoplasma-specific antibodies in patients suspected to have active toxoplasmosis: A cross-sectional survey.

BACKGROUND: The aim of this study was to investigate the presence and distribution of anti-toxoplasma-specific IgM and IgG tantibodies in patients suspected to have toxoplasmosis and investigate for any association between IgM and IgG antibodies and some toxoplasmosis risk factors as well. MATERIALS AND METHODS: In a comparative cross-sectional study, 70 patients suspected to had active toxoplasmosis and 30 control volunteers, who gave informed consent, entered the study. In each group, patient age, sex, signs of appearance, education level, residency status (urban / rural), occupation, frequency of toxoplasma-specific IgG and IgM antibodies, abortion history, and some risk factors (Direct cat exposure, Occupational exposure to raw meat, and Raw vegetable consumption) were recorded. The enzyme-linked immunosorbent assay (ELISA) kits (EUROIMMUN(®), United Kingdom) were used for the evaluation of anti-toxoplasma IgG and IgM antibodies according to the manufacturer's instructions. All analyses were done using SPSS-20. RESULTS: The frequency of toxoplasma-specific IgG and IgM antibodies like: Direct cat exposures, Occupational exposure to raw meat, and Raw vegetable consumption were not statistically significant between the two groups (P > 0.05). The history of previous abortions in women in the toxoplasmosis-suspected group was significantly higher than that in the controls (31.4% versus 6.7%; P = 0.009). CONCLUSION: The frequency of specific IgM and IgG antibodies in toxoplasmosis suspected in the toxoplasmosis and control groups was not statistically significant.

Scolicidal effects of squash (Corylus spp) seeds, hazel (Curcurbia spp) nut and garlic (Allium sativum) extracts on hydatid cyst protoscolices.

BACKGROUND: Because there is no effective drug therapy for hydatid cyst yet, assessment and finding of some new agents especially from herbal origin with a desired scolicidal effect attracts great attention for treatment and pre-surgical use to prevent the hydatid cyst recurrence. Hazelnut, squash seeds and garlic chloroformic and hydro-alcoholic extracts' scolicidal effects were examined. MATERIALS AND METHODS: Suspension of protoscolices was obtained from infected liver and or lung of sheep and goats from Ziyaran abattoir. The chloroformic and hydro-alcoholic extracts from hazelnut, squash seeds and garlic were extracted using the succilate method. Scolicidal effect of each extract assessed in different concentrations and effected time using microscopy and 0.1% eosin solution stained only killed protoscolices. RESULTS: Present study showed that garlic had more potent scolicidal effects among all the 3 plants and the chloroformic extract of garlic was the most potent protoscolicid among all of the extracts and killed 98% of protoscolices in 50 mg/ml on a minimum of 20 minutes exposure. CONCLUSION: Garlic chloroformic extract is a safe and potent protoscolicid and might be used in hydatid cyst treatment and pre-surgery to prevent secondary cyst recurrence.

Identification of genotypes of Giardia duodenalis human isolates in Isfahan, Iran, using polymerase chain reaction - Restriction Fragment Length polymorphism.

BACKGROUND: Giardia duodenalis is one of the most prevalent intestinal parasites of human. It also infects a wide range of mammals. Two genotype of G.duodenalis (A and B) were commonly reported among humans with different frequency of distribution in different geographical locations. This work was conducted to discriminate genotypes of Giardia duodenalis human isolates in Isfahan city using PCR- RFLP. This is the first molecular study on human isolates of G.duodenalis in the area. METHODS: Samples were collected from different health centers of Isfahan city during June 2011 and February 2012. From 175 Giardia positive stool samples 67 specimens were selected randomly. Cysts of Giardia positive samples were concentrated by flotation sucrose. Extraction of genomic DNA from trophozoite and cysts was performed using QIAamp Stool Mini kit with a modified protocol. PCR- RFLP method was used to amplify a fragment of 458bp at the glutamate dehydrogenase locus, and restriction enzymes BspLI and RsaI differentiated human genotypes A and B and their subgroups. RESULTS: PCR - RFLP assay of 67 isolates showed 40(59.7%) isolates as Genotype A group II, 23(34.32%) samples as Genotype B Group III and two (2.98%) sample as Genotype B group IV. Mixed genotype of (AII and B) was detected only in two isolates (2.98%). CONCLUSIONS: PCR - RFLP assay targeting gdh locus is a sensitive tool and discriminates genotypes, sub genotypes and mixed type of G.duodenalis. Results of our study suggest both anthroponotic and zoonotic origins for the infections respectively.

Do mutations in Plasmodium falciparum dihydropteroate synthase and dihydrofolate reductase confer resistance to sulfadoxine-pyrimethamine in Iran?

The key codons of dihydropteroate synthase (dhps) and dihydrofolate reductase (dhfr) genes implicated in sulfadoxine-pyrimethamine (SP) resistance were determined by allele-specific polymerase chain reaction in Plasmodium falciparum isolates collected in 1999 from 35 Iranian patients treated with SP. Seven isolates had Glu-540 dhps allele but 5 of these isolates were characterized to possess wild-type dhfr alleles. Seven additional isolates were polyclonal with mixed Lys- and Glu-540. The key dhfr mutation associated with pyrimethamine resistance, Asn-108, was found in 4 isolates. In one patient the presence of Lys- and Glu-540 in dhps and Asn-108 and Arg-59 in dhfr was associated with treatment failure. However, more studies are needed to determine whether clinical response to SP and mutations in these genes are correlated in Iran.