Jundishapur Journal of Microbiology
Volume:4 Issue: 3, Jul-Sep, 2011

Introduction and Nowadays, widespread application of antibiotics results in resistant microorganisms all over the world. Thus screening researches for the products with antimicrobial activity have been lead to probe natural sources of antimicrobial agents to find new pharmaceutical products. Here we describe the antagonistic activity of a bacterium isolated from digestive system of Iran endemic slug, Parmacella iberica, its biochemical identification and phylogenetic relationships and extraction of antimicrobial substance. Samples from digestive system of slugs were immediately transferred to broth medium. Antimicrobial agent-producing bacteria were isolated using serial dilution method on nutrient agar. One of them was selected for more investigation. Disk diffusion test was used against some laboratory standard strains to screen the antimicrobial potential. Identification of bacteria was done with 16S ribosomal DNA amplification and standard biochemical tests. Phylogenetic analysis of 16S rDNA gene of this bacterium showed similarity to Pseudomonas aeruginosa confirmed by the result of biochemical tests. The results of the antibiogram assay implied that the antimicrobial substance is broad-spectrum. This substance is nonpolar since it is extracted by different solvents. The cell growth measurements revealed that this compound was produced at first logarithmic phase. High antimicrobial effects of the bacterium are evident. Due to increscent need to new antibiotics, purification and identification of this antimicrobial compound seem to be necessary for introducing new medicinal source. Significance and impact of the study: The isolation of bacterial strain producing antimicrobial agent with the broad spectrum antimicrobial properties is proposed.

Introduction and Chronic infection with hepatitis C virus (HCV) is a major cause of cirrhosis and hepato-cellular carcinoma. Since response to anti-viral therapy in sufferers depends on HCV genotypes, determination of such genotype is of great significance to the treatment. This study seeks to estimate the HCV genotype prevalence in Shiraz, southern Iran and help specify the treatment course. A RT-PCR available kit with genotypes specific primer sets for major HCV genotypes (1a, 1b, 2 and 3a) was used. These primers amplified different parts of 5' un-translating region-core region of HCV genome. Genotyping test was performed for 634 patients with positive qualitative RT-PCR results. Of the 634 studied participants, 550(86.8%) were male and 84(13.2%) were female. Two hundred fifty nine (40.9%) of them were infected with 3a, 137(26.2%) with 1a, 55(8.7%) with 1b and 15(2.4%) with genotype 2 of hepatitis C. Mixed infection was found in 12 patients [1a+3a in 5(0.8%), 1a+1b in 4(0.6%) and 1a+2 in 3(0.5%)]. The extracted nucleic acid from 156(24.6%) samples did not react to the primer sets. This might be due to the presence of a genotype other than the above, or no sufficient copy of the virus. The results revealed the highest level of infection belonging to 3a followed by 1a. Since a considerable proportion of chronic hepatic C infected patients were intravenous drug abusers in the region; genotype 3a appears to be more prevalent among this group.Significance and impact of the study: The present study determined some critical information about the distribution of HCV genotypes in southern Iran.

Introduction and Asymptomatic bacteriuria (ASB) is the presence of significant numbers (≥105CFU/ml) of bacteria in the urine accompanying. The occurrence of ASB is widespread among elderly population. This study detected the prevalence of ASB in the elderly referred to the Taleghani hospital in Abadan, Iran. A total of 265 outpatients female and male aged between 60-90 years old who attended of Taleghani hospital in Abadan south western of Iran with no dysuria, frequency and urgency, fever, chills and flank pain, and were screened by collecting mid-stream urine samples for identification of the asymptomatic bacteriuria. The bacteria were isolated and disk diffusion method was used to study their patterns of antibacterial susceptibility. Asymptomatic bacteriuria was observed in 10.9% cases with predominant prevalence in female. A total of 29 isolated bacteria, Staphylococcus saprophyticus was considered as the most frequent organism which was accountable for 34.5% of the cases of ASB, and followed by Escherichia coli (31%), Klebsiella pneumoniae (17.2%), Proteus mirabilis (6.9%), S. aureus (6.9%) and Pseudomonas aeruginosa (3.4%). The resistance of the isolated bacteria to cephalotin and nitrofurantion were 78.3 and 69.1%, respectively. Most of the bacteria were sensitive to cefixime and ofloxacin. Nearly 11% of elderly person were identified to suffer from asymptomatic bacteriuria, and S. saprophyticus was a common pathogen. Significance and impact of the study: The elderly over 60 years has substantial risk of suffering from ASB, therefore they indispensable an accidental perform usual urine culture.

Introduction and Directed evolution is an incredibly powerful tool for altering the properties of enzymes. This approach uses a “sloppy” version of PCR, in which the polymerase has a fairly high error rate to amplify the wild-type sequence. The aim of this study was to produce the Bacillus subtilis mutant lipase which can be active at low temperatures using the error-prone PCR method. The pGEM-T vector containing B. subtilis lipase gene was transformed into the Escherichia coli DH5α. The lipase gene was mutated using the error-prone PCR technique. Screening of the mutants was carried out using both microtiter and Luria-Bertani plates containing 1% tributyrin and 100mg/ml ampicillin. Ten out of 1300 clones which showed the lipase activity at 10ºC were isolated. Among them, three desirable mutants named pFJK1, pFJK2 and pFJK3 were selected. Clones containing the wild type and the mutated lipase gene were both purified and characterized. The enzyme assay at different temperatures was carried out. The optimum temperature for the activity of lipase was achieved at 10ºС and 30ºС for the mutants and control, respectively. The relative enzyme activity of pFJK1, pFJK2 and pFJK3 at 10ºC was more than two times of their activity at 30ºC. The enzyme retained 70% of its activity at 10ºС and there was a drastic decline in enzyme activity at 50ºС. Based on the present findings, the enzyme was characterized as a cold adapted lipase which showed the highest activity at 10ºС.Significance and impact of the study: Cold adapted lipase can be developed for industrial applications such as additives in detergents.

Introduction and Today mining operations in the metallurgy generate secondary products which cannot be used directly in the basic technology. The mineral industry is increasingly faced with the need to process low grade ores and waste from current mining operations. Heap bioleaching of low grade sulfide is a developing technology that has been applied to the extraction of metal from secondary sulfide minerals. The microorganisms used in the bioleaching of the low grade sulfide ores are mostly different strains of Thiobacillus. The resistance of indigenous Acidithiobacillus ferrooxidans to metals such Cd+2, Co+2 and Zn+2 was investigated in the process of bioleaching tailing and low grade ore. Iron-sulfur oxidizing bacterium, At. ferrooxidans, has been isolated from pregnant leaching solution (PLS) of the Sarcheshmeh copper mine. At. Ferrooxidans was isolated, then it’s resistance to some heavy metals was investigated with shaking flasks at 150rpm and 30°C. The results indicated indigenous At. ferrooxidans tolerated high levels of Zn2+ about 30 (g/L) concentration, but it had less tolerance to Cd2+ and Co2+ (10g/L Cd2+ and 110ppm Co2+). This experiment demonstrated that Zn+2 toxicity is less than Cd+2 and Co+2 for isolated At. ferrooxidans. For this reason, isolated At. ferrooxidans can be used for bioleaching of tailing with high amount of zinc and waste with relative amount of cadmium.Significance and impact of the study: Using of isolated At. ferrooxidans in bioleaching of low grade sulfide ore which can tolerate presence of other heavy metals in mineral concentrate.

Introduction and Toxoplasma gondii is an intra cellular protozoan parasite which infects 30 to 60% of the world population. A wide range of toxoplasmosis conditions has been studied, but there are still unknown damages which must be explored. In the present research, the effect of this parasite on testosterone and cortisol changes in the infected men and women was examined. A total of 180 patients (73 females and 107 males), were examined for lgG anti-Toxoplasma antibody, cortisol and testosterone in their plasma. In addition, the patients also filled in personality questionnaire DASS21 (depression, anxiety, and stress). Results showed that 24(%13/33) females and 39(%20) males were positive with lgG anti-Toxoplasma antibody, respectively. A statistically significant correlation between Toxoplasma infection and testosterone and cortisol increase in women and men were observed. Stress and anxiety index also increased in men and women whereas depression index increased only in men. The results show a direct relation between raise Toxoplasma infection and cortisol, and testosterone increase in bout men and women and high DASS21 test score in Toxoplasma infected group. Significance and impact of the study: Our results may indicate that following Toxoplasma infection, the probability of stressful condition is high.

Introduction and Otitis externa, commonly called ear disease, is characterized by inflammation of the external ear canal. The aim of this study was to evaluate the antimicrobial potential of Cassia fistula flowers, leaves and bark extracts against Staphylococcus aureus, Proteus mirabilis, Escherchia coli, Pseudomonas aeruginosa, Acinetobacter sp., and Candida albicans, pathogens causing otitis externa and their comparison with locally available ear drops. Methanol, ethanol, acetone, aqueous (hot and cold) extracts from the flowers, leaves and bark of C. fistula were tested for their antimicrobial activity against six ear pathogens causing otitis externa determined by agar well diffusion method. Organic flower and bark extracts displayed activity against all tested ear pathogens whereas leaf extract showed activity against four tested bacteria and aqueous extracts were unable to exhibit any antimicrobial activity. Of the three organic solvents evaluated, acetonic flower extract was found to be best against S. aureus followed by bark extract and leaf extract. The acetonic flower extract showed larger inhibition zones compared to the herbal ear drops with minimum inhibitory concentration (MIC) of 6.25mg/ml. Acetonic extract of C. fistula flower may be used to treat otitis externa especially caused by S. aureus. However, more detailed studies such as in vivo testing and pharmacokinetics properties are needed to determine its therapeutic potential. Significance and impact of the study: Due to antimicrobial effects of amaltas organic extracts on ear pathogens, use of flower acetonic extract in herbal ear drops is recommended.

Introduction and Cryptosporidium parvum is an intracellular, extracytoplasmic, zoonotic, coccidian parasite that infects the microvillus regions of the digestive tract, and respiratory tract and causes lethal disease mainly among individuals suffering from immune deficiency and children. Suitable animal models are needed to identify different aspects of Cryptosporidium infection and the means of treating and preventing this infection. This study was conducted to represent a simple and usable animal model for C. parvum so that it can be used in the drug-related studies. Forty neonatal BALB/c mice and forty neonatal rats as group I (case) were inoculated with 5×105 oocysts. At the same time the animals in group II (control) received only sterile PBS solution. The intensity of infection was determined through the microscopic observation of the homogenized intestines of all the samples at 6, 9, 12, and 16 days of post infection. In mice and rats, the intensity of infection in group I (case) reached its peak on the 9th day after infection, then it reduced gradually and on day 16 post infection, it showed its lowest point. Meanwhile the non-infected animals in group II (control) had no detectable oocysts. As the intensity of infection in both models showed a similar pattern; and because of rat physiological similarity to humans and easier handling and manipulation during the procedure the rat model was considered experimentally preferable.Significance and impact of the study: This study introduces an indigenous laboratory animal model which can be used in cryptosporidium-related investigations and in vivo oocysts reproduction and it has no precedence in Iran.

Introduction and The soils are the major source of fungi. Some of the soil fungi can be pathogen to humans and animals. Rich keratinous materials in soil are the most reason for the incidence and growth of keratinophilic fungi. The purpose of this survey was isolation of keratinophilic and dermatophytic fungi from the soil samples which can be pathogen to humans and animals. Forty-five soil samples were collected from three different locations in Ahvaz, Khuzestan province, Iran. The isolated fungi were identified after slide culturing according to studies of their gross and micro-morphological features. Totally 644 colonies from soil samples collected from three different locations in Ahvaz. The dermatophytes isolated were identified as: Microsporum gypseum 3(0.47%) and Trichophyton mentagrophytes 2(0.31%). The most common non-dermatophytes fungi isolated were: Penicillium sp. 250(38.81%), Yeasts 115(17.86%), Fusarium sp. 81(12.58%), Chrysosporium sp. 57(8.85), Aspergillus flavus 34(5.28%), Mycelia sterilia 32(4.97%) and A. niger 25(3.88%). Our research shows the keratinophilic fungi and related dermatophytes present in the soils of Ahvaz, Iran. Significance and impact of the study: Survey results revealed the occurrence of keratinophilic fungi in soils of different locations in Ahvaz.

Introduction and Naphthalene as a cyclic aromatic hydrocarbon is one of the major part of pollution in oil-contaminated environments. Methyl tert-butyl ether (MTBE), an additive to gasoline, is widely used. MTBE is highly water soluble, toxic and causes cancer. Naphthalene degraders have gene for co-oxidation of other toxic compounds therefore Naphthalene degrader could utilize and oxidize other toxic contaminant. The aim of this study is to isolate and identify bacteria that could degrade Naphthalene and MTBE as a circular and liner component respectively. Freshwater was contaminated with Naphthalene and after several subcultures, strain SA86 was isolated and identified as Pseudomonas by biochemical test. Then bacterium was grown in culture with determined amount (1000ppm) of Naphthalene and the disappearance of Naphthalene was monitored with Gas Chromatography (GC) after six days. Then inoculate of the above bacteria was transferred to Teflon cape tube containing 500ppm MTBE and incubated for 24h. The removal of MTBE was measured by GC. The results showed that strain Pseudomonas (SA86) can degrade 99% of Naphthalene during six days incubation period. In addition this strain can remove 70% of MTBE. Meanwhile by products resulted from oxidation of MTBE were not observed. The results emphasise that Naphthalene degrading bacterium isolated in this study can remove MTBE as well as Naphthalene, while there are different in their chemical structures. Obviously Naphthalene induced the MTBE oxidation enzymes. Significance and impact of the study: Usually co-metabolism reaction is necessary for MTBE oxidation. However, here it is shown that Naphthalene grown cells are useful for MTBE removal with no co-metabolism reaction.

We report a 24-year-old woman with a Mycoplasma pneumonitis associated with subacute meningoencephalitis and acute cerebellitis that caused a cerebellar atrophy. Electroencephalogram showed diffuse dysfunction in the brain. There was few white blood cell but normal glucose and protein in the cerebrospinal fluid (CSF). Brain MRI showed bilateral atrophy of cerebellum.