Jundishapur Journal of Microbiology
Volume:4 Issue: 2, April-June 2011

Hepatitis D Virus (HDV) infection is a widespread disease that has affected a large number of population with hepatitis B Virus (HBV) infection in Iran. Disease is considered to be a major public health problem in Iran. Delta hepatitis is the least common form of chronic viral hepatitis and is the form most likely to lead to cirrhosis. Delta hepatitis is serologically complex, so effective therapy is difficult. The diagnosis is made on the basis of the presence of antibodies against HDV (anti-HDV) and hepatitis B surface antigen (HBsAg) in the serum of a patient with chronic liver disease. It is confirmed by the presence of the HDV antigen in liver or HDV RNA in the serum (by reverse-transcriptionpolymerase-chain-reaction assay). It is important to determine whether delta hepatitis is present because the responses to therapy of patients with this disease are less satisfactory than those with hepatitis B, and the recommended regimen of interferon alfa is different. The optimal treatment of HDV is uncertain. Thus, patients should ideally be treated as part of a clinical trial. The only treatment approved for chronic HDV is interferon alfa. Treatment should be administered for one year; whether longer duration of treatment will improve response rates remains to be established. Available data have not demonstrated an advantage from the addition of a nucleos/tide analogue.

Introduction and Klebsiella pneumoniae is an opportunistic pathogen most frequently associated with extended spectrum β-lactamase (ESBL) production. These organisms are usually resistant to most antibiotics and pose a serious threat for health care associated infections. Plant essential oils rich in carvacrol and thymol have gained importance for their antimicrobial activity. We determined the composition of Zataria multiflora essential oil of the Jandagh area in Iran and measured its activity against ESBL producing urinary isolates of K. pneumoniae. Essential oil was prepared from Z. multiflora at full flowering stage by hydrodistillation and its constituents were analyzed by a combination of capillary GC and GC-MS. Antibacterial activity was measured against 10 ESBL producing urinary isolates of K. pneumoniae as well as six ATCC bacterial standards by disc diffusion, minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) using broth microdilution. Zataria multiflora essential oil contained 25 constituents of which the major components were carvacrol (50.57%), thymol (13.38%) and p-cymene (8.27%). All tested bacteria were susceptible to the essential oil with the exception of Pseudomonas aeruginosa. Disc diffusion results showed inhibition zones of 18.3-30.3mm for the ATCC standards and 20.7- 29.7mm for the 10 clinical isolates. MIC and MBC values were 0.015- 2.0mg/ml for ATCC strains and 0.03 to 0.5mg/ml for the clinical isolates. Zataria multiflora may have the potential to be used against multidrug resistant organisms such as clinical isolates of ESBL producing K. pneumoniae.

Introduction and Oropharyngeal candidiasis is a relatively common mycotic infection in cancer patients. In vitro susceptibility of oropharyngeal Candida isolates can be useful in selecting the appropriate treatment for the best therapeutic outcome. The aim of this study was to evaluate the in vitro antifungal activity of Candida species against antifungal agents. In vitro activities of four antifungals were detected in 69 Candida isolates recovered from cancer patients in four university hospitals using the microdilution method described in the CLSI M27-A3 guideline. Only 12(17.4%) of Candida isolate were resistant to antifungal agents. Three isolates (4.4% included C. albicans, C. glabrata, C. tropicalis, and C. pelliculosa) were resistant to amphotericin B, 5(7.2% included two C. albicans, two C. glabrata, and one C. kefyr) were itraconazole resistant. Two (2.9% include one C. albicans and one C. glabrata) were fluconazole resistant. Caspofungin resistance was detected in two C. infanticola strains which were reported as a clinical isolate for the first time. All Candida isolates (n=69) taken together gave minimum inhibitory concentration (MIC90) value for amphotericin B, fluconazole, itraconazole and caspofungin of 1, 0.25, 32 and 0.25μg/ml, respectively. In total, 18.7% of C. glabrata and 7.8% of C. albicans isolates were fully resistant to both itraconazole and fluconazole. Caspofungin had activity against oropharyngeal non- albicans Candida species isolates, particularly against those with reduced susceptibility to amphotericin B, fluconazole and itraconazole.

Introduction and Tannin acyl hydrolase (tannase E.C. 3.1.1.20), commonly referred to as tannase, hydrolyses the ‘ester’ bond (galloyl ester of an alcohol moiety) and the ‘depside’ bond (galloyl ester of gallic acid) in substrates such as tannic acid, methylgallate and m-digallic acid. This is an inducible enzyme produced by various filamentous fungi. The aim of present research was to isolate, clone and sequence partial gene of tannase from Aspergillus niger. Aspergillus niger was grown in a selective medium and then genomic DNA was directly extracted from fungal mycelium. Using designed primers, PCR carried out and a 950bp expected band obtained that was ligated into cloning vector and then cloned into Escherichia coli Top-10F´. In order to screen transformed cells, the blue/white bacterial colony selection was carried out and expected the inserted DNA was extracted from putative transformants cells. Extracted DNA was sequenced using ABI automatic system and a 908bp intronlees motif was obtained using BlastX analaysing software that showed 36% identity with tannase precursor of A. oryzae and 29% with Debaryomyces hansenii tannase gene in amino acid level. In conclusion, the identification, isolation, cloning and sequencing of a 908bp partial sequence of tannase gene from A. niger which is considered as an important bioreactor and industrial fungus were reported.

Introduction and The potential risk of application of high dosage of traditional medicinal plants has not been fully understood. Appropriate microbial biosensors have been constructed for monitoring the toxicity of many harmful chemical compounds. The aim of this research was to see how effective are the different concentration of two medicinal plants extracts (Crocus sativus (saffron) and Cannabis sativa) on a bioluminescent marker system indicating their side effects. The stability and light intensity of Escherichia coli SM10 λpir were previously characterized and confirmed. Several concentrations of saffron and cannabis water extracts were prepared. The light intensity was measured for a mixture of 450µl of aqueous saffron extract and 50µl of biosensor using a luminometer. Results showed gradual decrease on light output in the way that luminescence decreased from 538859 RLU/s for 0.001g/ml aqueous saffron extract to 4830 RLU/s for 0.2g/ml concentration. Although induced increase in bioluminescence was observed for low concentration (0.001 and 0.01 v/v) of cannabis extract 0.25 and 1v/v concentration showed significant decrease in bioluminescent activity. Calculation of % INH of luminescent indicated the correct sensitivity of luminescent biosensor E. coli SM10 S1 to various concentration of saffron and cannabis extracts. The results show the appropriate interaction of constructed biosensor to different concentrations which can be used for further investigation on other ranges of concentrations. Application of luminescent microbial biosensor for investigation of the quality of products such as saffron and cannabis is new.

Introduction and

The uses of traditional medicinal plants for primary health care have steadily increased worldwide in recent years. Triphala has been used in the traditional medicine for the treatment of variety of diseases and therefore it becomes immense to study the phytochemical compounds and antibacterial activities. Aqueous and alcoholic extracts of both Triphala and its individual components were used, to evaluate antimicrobial activity.

Phytochemical (phenolic, flavonoid and carotenoid) and antibacterial activities of aqueous ethanolic extracts of Triphala and its individual components (Terminalia chebula, T. belerica and Emblica officinalis) were tested against several bacterial isolates. Isolates were recovered from urethral swabs, seminal fluid, urine, high vaginal swabs, skin swabs, blood, and sputum specimen of HIV infected patients.

Terminalia chebula has high phytochemical content followed by T. belerica and E. officinalis. In anti-bacterial activity, most of the bacterial isolates were inhibited by the ethanolic and aqueous extracts of T. chebula followed by T. belerica and E. officinalis in both disk diffusion and minimum inhibitory concentration (MIC) methods. But as a whole, Triphala did not show antibacterial activity. MIC of aqueous and ethanolic extracts of Triphala and its individual plant components were observed to vary from 0.1-100µg/ml.

In conclusion, this study showed that both ethanolic and aqueous extract of Triphala has potent antibacterial action against the wide variety of bacterial isolates from the HIV infected patients.

Introduction and Brucellosis is a zoonosis disease among animal and human, and has been endemic in Iran. The most important virulence factors of Brucella are related to their capability of intraphagocytic survival. Because of the side effects of brucellosis treatment regime, it is necessary to find new antimicrobial agents. The hop plant (Humulus lupulus) extract reported as having antimicrobial effects. The present study evaluated the antimicrobial activity of hops extracts against Brucella abortus 544 and B. melitensis 16M. Hops extracts were prepared in water, acetone and ethanol. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for two strains of Brucella determined by broth macrodilution and agar well diffusion methods. Effect of extracts on intramacrophge surviving of Brucella strains studied on cell culture of mouse peritoneal macrophages. Results indicated that MIC for aqueous extract was 1:80 (0.625mg/ml) and for acetonic and ethanolic extracts was 1:160 (0.05mg/ml). MBC for aquatic extract was 1:40 (1.2mg/ml) and for two other extracts was 1:80 (0.1mg/ml). In cell culture results indicated that all of extracts were effective and eradicated intramacrophage Brucella strains in 1:40 (1.2mg/ml for aqueous and 0.2mg/ml for ethanolic and acetonic extracts), 1:80 (0.625mg/ml for aqueous and 0.1mg/ml for ethanolic and acetonic extracts) and 1:160 (0.312mg/ml for aqueous and 0.05mg/ml for ethanolic and acetonic extracts) of extracts dilutions after 24h. Overall this study indicated that aquatic, acetonic and ethanolic extracts of hops showed antimicrobial effect against B. abortus and B. melitensis, therefore, they are useful in the treatment of brucellosis.

Introduction and Extremely halophilic Archaea belonging to the order Halobacteriales had been isolated from various hyper saline environments such as the Dead Sea, the Great Salt Lake, Sabkhas Lake, and natural or artificial Salterns. The main aim of this study was to isolate and characterize Halobacterium salinarum strains from samples of saline soils and lake water collected from four distinct regions of Iran. Saline soil samples and lake water samples were collected from different zones of Iran including a high salinity lake in Qom and Fars and also Seawater of Oromiea Lake and Persian Gulf. For isolation of H. salinarum from collected samples, Halobacterium salinarum medium were used. Isolates were identified by biochemical tests such as motility and morphological features, then identification confirmed by molecular characterization. Optimum NaCl requirement, pH and temperature were determined. The obtained results showed two isolates were motile, rod, Gram-negative, oxidase and catalase positive, with red pigmentation which grew in the presence of 25% NaCl. Isolates could hydrolyze casein and gelatin. The result of molecular characterization showed that two strains were H. salinarum. Basic local alignment search tool (BLAST) analysis showed 99% homology between these strains and H. salinarum strain R1. In this investigation we succeeded to isolate and identify H. salinarum in Iran saline ecosystems. There is a large diversity among halophilic bacteria in Iran ecosystems and isolation of H. salinarum from saline lakes will provide better information about halophilic bacteria in Iran ecosystems.

Introduction and Human astrovirus (HAstVs), belonging to a family of non enveloped, icosahedral RNA viruses and causes gastroenteritis both in infants and adults. The aim of this study was to determine the relative frequency of viral gastroenteritis caused by astrovirus among children less than five years referred to Ahvaz Aboozar hospital. Astrovirus infection was detected with Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). The authenticity of PCR products was confirmed by sequencing. Astrovirus infection was detected in 29 cases of 184 (15.77%), 13 positive samples belonged to the children up to six months. The relationship between gender and the prevalence of astroviral gastroenteritis was not significant. Most cases occurred during the coldest months of the year. After the sequencing, genotypes eight and four were the dominant types in this study. It was shown that human astrovirus plays an important role in gastroenteritis in Ahvaz, south west of Iran. The prevalence of the infection was very high. To decrease prevalence of astroviral infection, education and personal hygiene is advised.

Children with urinary tract abnormalities are susceptible to bacterial urinary infection (UTI); fungal infection, although rare, is reported to be increasing. Here, we describe a seven month-old male infant with posterior urethral valves who had developed both bacterial and fungal (Candidal) urinary tract infections. Direct and culture examinations of urine sample confirmed urinary tract candidiasis. Urine culture yielded more than 3×104 Candida albicans. Fluconazole started in the dose of 4mg/kg/day, single dose daily (25mg or 1/2 tablet) for two weeks. After two weeks antifungal therapy, urine sample was negative for C. albicans in both direct and culture examination. We concluded that fungal infection should be considered in the children with obstructing urinary tract abnormalities because its diagnosis may be missed.