Jundishapur Journal of Microbiology
Volume:3 Issue: 2, Apr-June 2010

Central nervous system (CNS) fungal infections have a high rate of morbidity and mortality that increased during last three decades. CNS fungal infections present a diagnostic and therapeutic challenge. High numbers of organ transplants, chemotherapy patients, intensive care unit hospitalizations, immunocompromised patients and haematological malignancies increase morbidity and mortality. Several fungi including, saprophytic fungi, melanized fungi, dimorphic fungi, yeast and yeasts-likes cause CNS fungal infections. New antifungal, posaconazole, voriconazole and echinocandins as well as traditionally antifungal, amphotericine B, flucytosine and itraconazole were used for CNS fungal infection therapy.

Introduction and Gastroenteritis, the inflammation of stomach and intestine, is caused by a variety of microorganisms including viruses. Especially adenoviruses type 40 and 41 of group F adenoviruses are two etiologies of gastroenteritis in newborns and infants of less than five years old. The aim of this study was to determine the prevalence of ad40 and ad41 inducing gastroenteritis in children hospitalized in Ahvaz Abuzar Hospital, Iran, during October 2007-2008. Fecal samples collected from the patients were tested first by an ELISA kit, specific for adenovirus detection. All samples including positive specimens by ELISA method were subjected to two rounds of PCR test. A specific pair of primers for adenoviruses 40 and 41 was applied PCR method. Out of 280 fecal specimens collected from diarrheic children, 7(2.5%) were positive by ELISA test and 12 (4.3%) by PCR method. All of positive samples belonged to ad41. From group F of adenoviruses, adenovirus 41 is the major etiology of gastroenteritis in Ahvaz area.

Introduction and The early secretory antigenic target 6kDa protein (ESAT-6) antigen from Mycobacterium tuberculosis is a dominant target for cell-mediated immunity in the early phase of tuberculosis (TB) in TB patients. ESAT-6 was found to distinguish TB patients from BCG-vaccinated donors. The aim of this study was cloning and expression of ESAT-6 of M. tuberculosis. DNA was extracted from M. tuberculosis H37Rv. PCR was performed using gene-specific oligonucleotide primers and the PCR products were inserted into the pET102/D vector and transferred into Escherichia coli strain TOPO10. The recombinant plasmids transferred into E. coli strain BL21. The transformed plasmid into E. coli strain BL21 was effectively expressed. The expressed fusion protein (23kDa on SDS-PAGE) was found almost entirely in the soluble form and the recombinant protein was purified by Ni-NTA column. We successfully cloned and expressed ESAT-6 protein of M. tuberculosis in E. coli. As a specific antigen, it can be useful for diagnosis of both active and latent tuberculosis with ELISA in future.

Introduction and Intestinal parasitic infection is an important problem in the Human Immunodeficiency Virus (HIV)-infected patients. The aim of this study was to investigate the prevalence of intestinal parasitic infections among HIV+ patients in Mashhad, Iran. A coproparasitological study was conducted from October 2005 to August 2006 at Emam Reza hospital, Mashhad University of Medical Sciences, Iran. It was carried out on 31 HIV+ patients admitted at the HIV clinic and 20 HIV-negative individuals as control group using direct and formalin-ether sedimentation concentration methods, trichrome and acid-fast staining. Overall prevalence of intestinal parasites among HIV+ population was 67.7% and in control group was 55% without significant difference between the two groups. More specifically, the following parasites were identified in HIV+ group: Giardia lamblia 22.6%, Blastocystis hominis 22.6%, Chilomastics mesnili 22.6%, Entamoeba coli 9.7%, and Entromonas 3.2%. In the control group Entromonas (45%), B. hominis (15%), E. coli (10%), G. lamblia (5%), and Hymenolepis nana (10%). However, the prevalence of G. lamblia, B. hominis and C. mesnili was greater for HIV+ patients (p<0.05). There were statistically significant differences between trichrome staining (28, 54.9% positive for parasites), acid fast methods (6, 11.8%), direct method (7, 13.7%) and formalin-ether method (13, 25.5%) in detection of parasites in two groups (p< 0.05). Our study shows the importance of testing for intestinal parasites in patients who are HIV-positive, and emphasizes the necessity of increasing awareness among clinicians regarding the occurrence of parasite infections in these patients. Routine examination of stool samples for parasitic infections could significantly benefit the HIV-infected individuals by contributing to reduce morbidity, mortality and improved quality of life.

Introduction and Otomycosis is a fungal infection of external auditory meatus. The acute form of the disease causes secretion and pruritus. The usual prescribed medicines for otomycosis are topical clotrimazole 1%, amphotericin B and otosporin. The aim of the present study was to evaluate the efficacy of treatment with isopropyl alcohol and acetic acid for otomycosis. In the present study 910 patients examined and those suspected to have otomycosis referred to medical mycology laboratory of Golabchi, Kashan. A questionnaire was also filled for each patient. Both direct and culture examinations were used to confirm otomycosis in the patients. Then the patients were treated with the mixture of isopropyl alcohol+acetic acid. Out of 910 examined patients, 60 patients were suspected to have otomycosis and referred to medical mycology lab. Mycological examinations confirmed otomycosis in 52 patients (86.7%). Most of the patients (78.8%) were cured perfectly after therapy with the mixture of alcohol and acetic acid. After three weeks, in addition to elimination of clinical signs further smear showed no sign of disease. However in four patients there was a relapse of the disease. Due to therapeutic effect of the mixture of isopropyl alcohol and acetic acid for otomycosis, its low side effects and low rate of relapse, it is recommended to use this mixture for the treatment of otomycosis.

Introduction and Caspases belong to a distinct class of cysteine proteases which also includes hemoglobinases, gingipains, clostripains, and separases, the proteases involved in chromosome segregation in mitosis and meiosis. Two families of predicted Caspase Homoglobinase Fold (CHF)-proteases were identified and shown to be more closely related to the caspases than to other proteases of this class and hence dubbed paracaspases and metacaspases. The aim of present study was to isolate genes particularly caspase-related ones up-regulated in the earliest stages of the stationary phase (up to four hours) and using clones to perform a PCR on cDNA prepared from exponential and stationary phase mRNA. After the use of the clorimetric assay to get evidence of caspase activity, both strands of cDNA were used for cloning the PCR products. The whole plasmid and freeze-dried material were sent for automated sequencing. By searching in NCBI homepage, particularly Blast part, all the alignment sequences were shown and then after similar genes were recognized as well. There was no sign of any cDNA in the midlog and 4h stationary phases which shows no RNA synthesis in those phases. The extended sequence of the gene was found to have a high level of identity (87%) with a metacaspase from Schizosaccharomyces pombe through a BLASTX search. This information may be used to fabricate microarrays which would enable a genome-wide analysis of gene expression of Aspergillus fumigatus as it enters and during the stationary phase.

Introduction and Oral metronidazole is reported before as effective treatment of cutaneous leishmaniasis. The aim of the present study was to evaluate the efficacy of intralesional metronidazole injection versus intralesional meglumine antimoniate (Glucantim®) in the treatment of cutaneous leishmaniasis. The 36 patients with clinical and parasitologic diagnosis of cutaneous leishmaniasis participated in this study. The patients were randomly divided into two groups. Group one, 18 patients were treated with weekly intralesional of Glucantim injections, and group two, 18 patients were treated with weekly intralesional injections of metronidazole. Intralesional injections was administered enough to blanch the lesions surfaces (0.5-2ml for each lesion in both groups). Twenty eight patients completed the study. Sixteen patients in group one and 12 patients in group two. In group one, 13 patients recovered with eight injections (81%). and of group two, only three patients recovered with eight injections (16.6%). The pain of intralesional injections of metronidazole was much more than the Glucantim injections. Intralesional metronidazole injections have little effect for the treatment of cutaneous leishmaniasis.

Introduction and Today methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase negative Staphylococci (MRCNS) are frequent causes of nosocomial infection. Extensive burn injuries, extended hospitalization and inappropriate antibiotic therapy have been identified as risk factors for MRSA and MRCNS carriage and infection. The aim of this study was to assess the prevalence of methicillin resistance among clinical isolates of Staphylococci taken from burn patients using four separated methods and also determination of susceptibility pattern to amikacin, ciprofloxacin, vancomycin, carbenicillin and gentamicin. A total of 185 clinical staphylococcal isolates from wound and blood specimens were evaluated for susceptibility to oxacillin using oxacillin and cefoxitin disk diffusion method, agar screening containing 6 microgram oxacillin /ml, oxacillin E test and polymerase chain reaction for predicting mecA gene. The results showed that 27.8% of wound and blood specimens were infected by Staphylococci and among these 60% were identified as methicillin resistant. We found no significant differences between the results of PCR assay and conventional disk diffusion method by oxacillin and cefoxitin disk, however the results of cefoxitin disk was more significant than oxacillin and gave better results. Both of the sensitivity and specificity value were similar (99%, 100%) for E test and agar screen test. Furthermore in E test for detection of minimum inhibitory concentration (MIC), more than 93% of MRSA and 15% of MRCNS isolates had MIC value more than 256µg/ml. We also determined a significant difference pattern between methicillin resistant and methicillin susceptible Staphylococci to five antimicrobial agents. In conclusion, our results showed that the prevalence of methicillin resistant Staphylococci in our center was very high and cefoxitin disk test is reliable alternation for detection of methicillin resistant Staphylococci.