In-vitro Activity of Trigonella foenum graecum Seeds Against a Clinical Strain of Acanthamoeba Genotype T4.

Acanthamoeba keratitis (AK) is a sight-threatening infection of the cornea disease that often presents with a lengthy and not fully effective treatment. Current therapeutic options against Acanthamoeba are not very effective against the cyst. Calibrated trophozoite/cyst suspension was incubated with the same volume of serial dilutions of the Trigonella foenum graecum aqueous extract (200, 250, 350, 450, 600, and 750 mg/mL) in microcentrifuge tubes and mixed by pipetting up and down. After that, the tubes were incubated at 26 ºC for 24, 48, and 72 h. The obtained result revealed that incubation of the extract (at concentrations ranging from 200 to 750 mg/mL) with Acanthamoeba was able to decrease the number of viable trophozoites and cysts. In the presence of up to 450 mg/mL non-viable trophozoites were observed whereas cysts were only eliminated when incubated with 750 mg/mL of the extract after 24 h. Furthermore, no cytotoxicity of the extract even at the highest concentration tested in the study showed to be toxic for corneal cells. Further studies should be carried out in order to elucidate the active compounds involved in the observed anti-Acanthamoeba activities which could be used for the development of novel therapeutic approaches against Acanthamoeba infections.

Occurrence of Naegleria species in therapeutic geothermal water sources, Northern Iran.

Potentially pathogenic Free-Living Amoebae include members belonging to Naegleria genus. The species N. fowleri is known worldwide as the causative agent of the lethal Primary Amoebic Meningoencephalitis (PAM). Only one clinical case of N. fowleri has been reported in Iran. Several species of Naegleria have been reported to be natural carriers of other potentially pathogenic microbial agents. The thermotolerance properties of this genus facilitates their presence in geothermal water sources including hot springs and spas. In the current study water samples were collected from 22 therapeutic hot springs, Northern Iran and investigated for the presence of Naegleria spp. using morphological keys and PCR/DNA sequencing based methods. Incubation of collected samples were done at both 30°C and 45°C in order to detect Naegleria spp. and N. fowleri, respectively. Thermotolerance assay and flagellation tests were also performed. The obtained results revealed that 54% of the investigated water samples were positive for Naegleria spp. including N. australiensis, N. americana, N. dobsoni, N. pagei, N. polaris and N. fultoni. The pathogenic N. fowleri was not detected. The most detected Naegleria was belonged to N. australiensis. This is the first report on the Naegleria spp. occurrence in hot springs in Northern Iran showing that most of the surveyed hot spring sources were contaminated with non-pathogenic Naegleria spp. However, due to the recent report of PAM in the country, further studies to investigate the presence of pathogenic N. fowleri in the environment and clinical samples is needed in the region and worldwide.

Frequency of Cutaneous Leishmaniasis and Species Identification in Suspected Individuals from Golestan Province, Northern Iran in 2014.

BACKGROUND: Leishmaniasis is a zoonotic disease caused by species of protozoa of the genus Leishmania. In recent years, incidence of cutaneous leishmaniasis has increasing trend in Golestan Province, North of Iran. The aim of the present study was to identify the frequency of cutaneous leishmaniasis using PCR-RFLP in patients referred to Kalaleh Health Center, during 2013-14. METHODS: This descriptive cross-sectional study was conducted on 70 individuals with suspected cutaneous leishmaniasis that referred to health center of Kalaleh County, Golestan Province, Northern Iran, from Sep 2013 to Nov 2014. Samples of cutaneous lesions were examined microscopically. DNA was extracted from all of the positive smears and PCR was done on ITS-1 gene. RFLP was performed using HaeIII enzyme for species identification. RESULTS: Totally, 38 out of the 70 (54.3%) suspected individuals including 22 males (57.9%) were found positive by microscopic examination. All of microscopically positive samples were confirmed to be positive for Leishmania DNA (approximately 340 bp bands were detected). RFLP revealed 140 bp and 200 bp bands (approximate size), indicative of L. major. CONCLUSION: The detected species of studied region was L. major. Cutaneous leishmaniasis has high prevalence in Kalaleh County, thus more studies on leishmaniasis in the animal reservoirs, comparison of homology of animal and human isolates and a survey regarding natural infection of vectors in this region is highly recommended.