Genotype Characterization of Human Hydatid Cyst Isolates From Patients in Karaj, Iran, Using COX1 Gene Sequence

Background: Cystic echinococcosis is a main zoonotic infection. It can cause serious clinical problems for human health around the world. Genotypic specification of Echinococcus granulosus in human is important due to control and prevention programs.Objective: In this investigation, genetic characteristics of human isolates of E. granulosus in Karaj, Iran, were studied.Materials and Methods: In this review, 3 isolates of surgically removed hydatid cysts were obtained from patients in Shahid Madani hospital, Karaj, Iran in 2014. DNA was extracted from the protoscolex of the cyst, and polymerase chain reaction (PCR) assay was done on the COX1 gene.Results: DNA fragments were sequenced and the results were aligned and analyzed. Among the isolates, 3 (100%) were E. granulosus (G1) strain.Conclusion: The G1 genotype was the most superior strain from human isolates of hydatid cyst in Karaj

Identifcation of Candida Species Isolated From Oral Colonization in Iranian HIV-Positive Patients, by PCR-RFLP Method.

Background: The incidence of opportunistic infections due to Candida albicans and other Candida spp. has been increasing. Rapid identifcation of candidiasis is important for the clinical management of immunocompromised patients. Polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) is a rapid, sensitive, and specifc method for detection of clinically important fungi. Objectives: The purpose of this study was to identify Candida spp. isolated from the oral cavities of HIV-infected patients in southeastern Iran (Kerman), by using PCR-based restriction enzyme digestion. Patients and Methods: We identifed 96 Candida isolates obtained from 139 Iranian patients infected with the human immunodefciency virus (HIV), between April 2009 and April 2010, by using PCR-RFLP assay. Universal primers for the internal transcribed spacer (ITS) region (ITS1–ITS4) of the fungal rRNA genes were used for this assay. Results: We successfully identifed the different Candida spp. by using the restriction enzyme MspI. C. albicans was the most commonly identifed species (82.2%), followed by C. glabrata (7.29%), C. parapsilosis and C. kefyr (both 4.1%), and C. tropicalis (2%). Conclusions: PCR-RFLP is a highly sensitive, specifc, and direct method for fungal detection and can be used for fungal epidemiological studies in HIV-positive and other immunocompromised patients

The assessment of growth inhibition effect of Copper chloride and 8-Hydroxyquinoline compound on Trichophyton verrucosum

Background: Dermatophytosis or ring worm is one of the most common fungal infections. Trichophyton verrucosum (T.verrucosum) is common among the agents that cause dermatophytosis. The aim of the present study was to evaluate of antifungal activity of copper chloride (cucl2) and 8- hydroxyquinolin compound on T.verrucosum. Methods: One standard strain and two clinical isolated of T.verrucosum were used in this study. The standardized method, M-38 A for filamentous fungi, was used according to the guidelines of National Committee for Clinical and Laboratory Standards (NCCLS) to determine MIC. The effect of CuCl2 and 8-Hydroxyquinolin investigated by using various concentrations of this compound in T.verrucosum cultures. Results: Cucl2 and 8-Hydroxyquinolin compound exhibited antifungal activity on the three mentioned strains and the MIC of this compound was 25μg/ml. Conclusion: Cutaneous The new antifungal drug scheming is required to discover new antifungal agent. The data showed that Cucl2 and 8-Hydroxyquinolin compound had antifungal effect, so they could be a good candidate for making of new antifungal compounds

PCR - RFLP patterns for the differentiation of the Fusarium species in virtue of ITS rDNA

Background and Purpose: The Fusarium species are among the most important fungi in the medical, veterinary and agricultural fields. Materials and Methods: In the present study, 172 strains of these fungi have been analyzed. The high molecular weight DNAs were extracted from 23 reference strains as well as from 149 isolated Fusarium species. Using the designed nucleotide primers from rDNA of Fusarium species, PCR analysis was performed for the amplification of ITS regions. Afterwards, the location of the effective endonuclease enzymes has been evaluated within approximately 930 bp of rDNA sequence. Results: Through the selected enzymes including; HhaI, MspI, TaqI and FaqI, the mentioned Fusarium species have been divided into 33 groups. The first three enzymes were able to classify Fusarium species into 23 groups of which 19 groups included one member, one group included two members and three groups included three members of the Fusarium species. This study also revealed the possibility in the identification of F. semitectum, F. solani complex, F. pseudograminearum, F. nisikadoi, F. coeruleum and F. acuminatum species by one unique enzyme. In addition, our study indicated the ability of the differentiation of F. Compactum from F. equiseti. Conclusion: As Compared to previous studies with more endonuclease enzymes and with limited in identifications, the ITS-RFLP patterns reported here an attempted to evaluate most of the Fusarium species successfully

Effect of biogenic selenium nanoparticles on ERG11 and CDR1 gene expression in both fluconazole-resistant and -susceptible Candida albicans isolates

Background and Purpose: Candida albicans is the most common Candida species (spp.) isolated from fungal infections. Azole resistance in Candida species has been considerably increased in the last decades. Given the toxicity of the antimicrobial drugs, resistance to antifungal agents, and drug interactions, the identification of new antifungal agents seems essential. In this study, we assessed the antifungal effects of biogenic selenium nanoparticles on C. albicans and determined the expression of ERG11 and CDR1 genes. Materials and Methods: Selenium nanoparticles were synthesized with Bacillus sp. MSH-1. The ultrastructure of selenium nanoparticles was evaluated with a transmission electron microscope. The antifungal susceptibility test was performed according to the modified Clinical and Laboratory Standards Institute M27-A3 standard protocol. The expression levels of the CDR1 and ERG11 genes were analyzed using the quantitative real-time polymerase chain reaction (PCR) assay. Results: The azole-resistant C. albicans and wild type C. albicans strains were inhibited by 100 and 70 µg/mL of selenium nanoparticle concentrations, respectively. The expression of CDR1 and ERG11 genes was significantly down-regulated in these selenium nanoparticle concentrations. Conclusion: As the findings indicated, selenium nanoparticles had an appropriate antifungal activity against fluconazole-resistant and -susceptible C. albicans strains. Accordingly, these nanoparticles reduced the expression of CDR1 and ERG11 genes associated with azole resistance. Further studies are needed to investigate the synergistic effects of selenium nanoparticles using other antifungal drugs. &nbsp

Identification of Candida species by molecular and mycological methods in pregnant women with vulvovaginal candidiasis

Background: Vaginal candidiasis is common in during pregnancy. It may lead to complications like abortions, premature birth, low birth weight, chorioamnionitis and fungal systemic neonatal infection. The aim of present study was identification of Candida species by mycological and molecular methods in pregnant women with vaginal candidiasis. Methods: This cross-sectional study was performed on 80 pregnant women with or without clinical symptoms of vulvovaginal candidiasis referred to Shahid Noorani Talesh Hospital, Gilan University of Medical Sciences, Iran, from April to December 2015 (8 months). All specimens were examined by direct microscopy and culture on CHROMagar Candida medium for isolation and differentiation of major clinical-significant Candida species (spp.). Cultured media were incubated at 35 &deg;C for 48 hours and evaluated based on color and number of grown colonies. If no growth was observed, the media were incubated for several additional days. Subcultures were done on Sabouraud dextrose agar (Merck, Germany) and Corn meal agar with Tween 80 media (Micromedia, Hungary) for further study. Identification of Candida spp. carried out by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: In this study, vulvovaginal candidiasis was observed in 20 (25%) patients. Twenty-two isolates were obtained from culture of specimens on CHROMagar Candida medium (Paris, France). The most common isolated species was Candida albicans 16 (72.8%) and followed by Candida glabrata 5 (22.7%), Candida tropicalis 3 (13.6%) and Candida krusei 1 (4.5%) cases. Two patients had mixed infection with 2 different Candida species (C. albicans and C. glabrata) While using PCR-RFLP method, the Candida species were identified as 13 (59.1%) Candida albicans, 5 (22.7%) Candida glabra, 3 (13.6%) Candida tropicalis and 1 (4.5%) Candida krusei cases, respectively. In direct examination were seen yeast budding cells and pseudohyphae in 8 culture positive specimens. In the present study, results of conventional mycological method in differentiation of Candida spp. were consistent with molecular results in 80% of cases. There was also significant correlation between vulvovaginal candidiasis with clinical symptoms (P<0.0001), including diabetes mellitus (P<0.014), and taking antibacterial drugs (P<0.003) in pregnant women. Conclusion: PCR-RFLP was able to identify correctly the Candida spp. as a complementary method

In Vitro Assay of Paecilomyces lilacinus Biocontrol Effects on Fasciola hepatica Eggs Illustrated in Scanning Electron Micrographs

Background: Fascioliasis is a zoonotic disease caused by the liver fluke Fasciola hepatica. Drug resistance, high costs of treatment and economic losses in meat production have emerged the need of alternative control measures into consideration. The aim of this study was to evaluate the in vitro ovicidal activity of Paecilomyces lilacinus fungus on F. hepatica eggs. Methods: P. lilacinus isolated from the soil of natural environment was challenged on F. hepatica eggs to observe the bio control effect of nematophagous fungi on trematode helminth eggs. The study was conducted in Tehran University of Medical Sciences, in 2015. Within 21 d of experiment, destructive effects exhibited on the eggshells were investigated using optical and Scanning Electron Microscopy. Results: The effective role of P. lilacinus on damaging the eggs of F. hepatica was noticed. Conclusion: This finding is promising for advantageous use of nematophagus fungi as a natural constituent in hyper endemic areas for certain helminthic infections like fascioliasis with diverse kinds of herbivores as egg passer hosts

Candida colonization and species identification by two methods in NICU newborn

Background: Over the last two decades invasive candidiasis has become an increasing problem in neonatal intensive care units (NICUs). Colonization of skin and mucous membranes with Candida spp. is important factor in the pathogenesis of neonatal infection and several colonized sites are major risk factors evoking higher frequencies of progression to invasive candidiasis. The aim of this study was to detect Candida colonization in NICU patients. Methods: This cross-sectional study was conducted on 93 neonates in NICUs at Imam Khomeini and Children Medical Center Hospitals in Tehran. Cutaneous and mucous membrane samples obtained at first, third, and seventh days of patients&rsquo; stay in NICUs during nine months from August 2013 to May 2014. The samples were primarily cultured on CHROMagar Candida medium. The cultured media were incubated at 35&deg;C for 48h and evaluated based on colony color produced on CHROMagar Candida. In addition, isolated colonies were cultured on Corn Meal Agar medium supplemented with tween 80 for identification of Candida spp. based on their morphology. Finally, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was performed for definite identification of isolated species. Results: Colonization by Candida spp. was occurred in 20.43% of neonates. Fifteen and four patients colonized with one and two different Candida spp., respectively. Isolated Candida spp. identified as; C. parapsilosis (n: 10), C. albicans (n: 7), C. tropicalis (n: 3), C. guilliermondii (n: 2), and C. krusei (n: 1). In present study non-albicans Candia species were dominant (69.56%) and C. parapsilosis was the most frequent isolate (43.47%). Using Fisher's exact test, the correlation between fungal colonization with low birth weight, low gestational age, and duration of hospital stay was found to be statistically significant (P=0.003). Conclusion: The results of this study imply to the candida species colonization of neonates. Neonates in NICU are at the highest risk for severe infection with Candida parapsilosis. Therefore, isolation of C. parapsilosis as the most common species (43.47%) in present study was noteworthy

Molecular Characterization of Cryptosporidium Isolates from Humans and Animals in Iran

Isolates of Cryptosporidium spp. from human and animal hosts in Iran were characterized on the basis of both the 18S rRNA gene and the Laxer locus. Three Cryptosporidium species, C. hominis, C. parvum, and C. meleagridis, were recognized, and zoonotically transmitted C. parvum was the predominant species found in humans