International Journal of Molecular and Clinical Microbiology
Volume:2 Issue: 1, Winter and Spring 2012

The aims of this study were to detect Helicobacter pylori (H. pylori) by culture, polymerase chain reaction (PCR) and histopathologic methods, to determine the prevalence of active H. pylori infection in Elazig Province, East of Turkey and to evaluate the relationship between H. pylori infection and sex. Antrum and corpus samples of 184 Turkish patients (85 male and 99 female, age range 17 to 92 years, average 49) with gastrointestinal complaints attending the Gastroenterology Department of Firat University Hospital during 2009 and 2010 were used in this study and examined for the presence of H. pylori using culture, PCR and histopathological examination. Patients were grouped as gastritis (G) in 155 cases, peptic ulcer (PU) in 26 cases, gastric cancer (GC) in 3 cases at the time of endoscopy. H. pylori was isolated in 61 (33.2%) samples. By PCR, H. pylori was detected in 140 (76.1%) patients, 115 (74.2%) cases with G, 23 (88.5%) cases with PU and 2 (66.7%) cases with GC. Fifteen of 155 patients with G were excluded from the histopathological evaluation due to inadequate material given. Histopathological examination of the 140 patients with G was detected to be H. pylori positive in 96 (68.6%). Further investigations are required to determine significant differences between tests used for detecting H. pylori.

Infectious bursal disease virus (IBDV) is the causative agent of Gumboro disease, an infectious disease of global economic importance in poultry. Structural protein VP2 of IBDV is the most frequently studied protein due to its significant roles in virus attachment, protective immunity, and serotype specificity. The objective of the present study was to improve the expression of hypervariable region of VP2 protein (hvVP2) in Escherichia coli (E.coli). The results showed that the hvVP2 was expressed in very low amount in E.coli. But, codon optimized hvVP2 protein showed significantly enhanced protein expression level. The coding sequence of hvVP2 was amplified and then identified by polymerase chain reaction (PCR) and sequencing. To achieve high-level expression of hvVP2 protein, we optimized hvVP2 gene base on E. coli preferred codons and synthesized the optimized gene. The synthetical gene was cloned into expression vector pET-26b and expressed in E. coli BL21 (DE3). After induction with Isopropyl-D-1-Thiogalactopyranoside (IPTG) and optimization the conditions of expression, the hvVP2 protein was relatively increased and identified by SDS-PAGE and Western blotting. Productive conformation can now be used for structure-based design purposes as well as structure-function relation of VP1 protein. It is suggested that the codon optimized hvVP2-His protein may be a useful option (but it is not enough) for developing diagnostic tests and immunization proposes.

Southern margin of the Caspian Sea due to keeping of domestic animals and abundance of surface water reservoirs is prone to infection with Leptospira sp.. This study was performed to determine anti-Leptospira antibody frequency in Golestan province, southeast of the Caspian Sea in the north of Iran. This study was performed on 1028 people in Golestan province. Sampling was stratified. Leptospira antibody (IgG) was measured with DRG (USA) enzyme linked immunosorbent assay (ELISA). 107 cases (10.4%) had anti-Leptospira sera antibody. Distribution of positive cases was 59 (10.9%) in women and 48 (9.8%) in men. Most positive cases were in the 35-44 years old group. The frequency of positive cases in the people who was living in the towns and villages in Golestan Province were 9.4% and 11.1%, respectively. No significant difference was seen between the groups. According to the data presented in this study and compared to the published data it was shown that the frequency of antibody against Leptospira in Golestan province is lower than both Guilan and Mazandaran provinces in the north of Iran.

Determination of the frequency of bacteria isolated from blood culture and the pattern of their antibiotic sensitivity is epidemiologically of great importance and can help choosing primary antimicrobial treatment. The aim of the current study was to determine the most common bacterial factors of blood infection and their antibiotic sensitivity in a one year period from October 2010 till the end of September 2011 in Imam Khumeini hospital of Ardebil. Data collected from total of 1469 patients susceptible to blood infection hospitalized in Imam Khumeini hospital and analyzed using SPSS (version 18) statistical software and investigated with descriptive statistics and frequency diagrams. Only values with P≤0.05 were considered as significant. The highest rate of bacteremia prevalence observed in emergency unit. Of 1469 blood sample, 91 (6.19%) were infected with bacteria. Infection prevalence reported higher in male (55.4%) than to female (44.6%). The most common bacteria isolated were, Negative-coagulase Staphylococcus (37.3%), Escherichia coli (22%), Staphylococcus aureus (11%) and Staphylococcus epidermidis (11%), respectively. The highest antibiotic sensitivity was associated with Gentamicin with the frequency rate of 42 (51.3%) and Ciprofloxacin with the frequency rate of 35 (42.7%). Gentamicin and Ciprofloxacin determined as two completely effective antibiotics on isolated strains. Regarding the increased prevalence of infection with Negative-coagulase Staphylococcus in the current study and other similar studies, the morbidity and mortality rate of the disease can be decreased using proper antibiotic treatments complying with the exact results of the disc diffusion test method.

Bacillary dysentery, or shigellosis, is a disease of humans in which the colonic epithelium is invaded by bacteria and subjected to inflammatory destruction. The aim of this study was to develop a polymerase chain reaction(PCR) test for detection of virulent Shigella spp.. For this purpose, the primers were designed to amplify a 526-bp internal region of the Shigella spp. icsA gene, which encodes IcsA (intracellular spread)/VirG protein, a 116-kDa surface exposed outer membrane protein that mediates actin polymerization to aid bacterial movement inside the cell. The use of PCR to amplify a specific icsA gene fragment serves as a highly specific and sensitive method to detect virulent bacteria of the genus Shigella. Specific DNA band was obtained by using isolated plasmid DNA of Shigella and a bacterial suspension. Amplification of extracted DNA from all other genera of the family Enterobacteriaceae and various other gram-positive bacteria yielded negative results. Therefore this PCR method, can serve as a routine protocol for detecting and identifying virulent Shigella spp. from clinical samples.

Nanotechnology is a new and specific method for therapy of diseases. Nanosilver particle is one of the functional fields of nanotechnology. It has been shown that these nanoparticles have antibacterial and antifungal properties. Different studied have been shown that nanoparticles may have important role in medicine. For example, it has been shown to have inhibitory effects on the treatment of many diseases. The shape and size of nanoparticles are very important. In this study the inhibitory effects of silver on E. coli has been investigated. Nanosilver in different concentrations on inseminated blank antibiogram disk was placed on cultivated nutrient agar environment by 0.5 Mac Farland`s standard. After the first, second, and eight day the diameter of disks was measured. It was found that nanosilver in a concentration of 50ppm is more effective than the other concentrations for growth inhibition of E. coli. The best time for induction of inhibitory effects on E.coli was 3 day post treatment with silver nanoparticles. Investigation of inhibitory effects for nanosilver concentration on E. coli is an example of functional effects of new nanobiotechnlogy. It was shown that eucalyptus and nanosilver has the maximum inhibitory effects on the growth of E.coli 1 day post treatment. The results from this study suggest that silver nanoparticles in combination with eucalyptus extracts may be useful for the therapy of human bacterial diseases.

Medicinal plants are a source of great economic value all over the world. Nature has bestowed on us a very rich botanical wealth and large number of diverse types of plants grows in different parts of the country. In this study the in vitro antimicrobial activity of crude ethanolic, methanolic and water extracts of the stem bark of O.basilicum were investigated. The extracts exhibited antimicrobial activities with zones of inhibition ranging from 5 to 12, 8 to 20 and 0 to 8 mm for ethanol, methanol and water extracts, respectively. The minimum inhibitory concentration (MIC) of the ethanol extract was between 0.5 and 6.25 mg/ml while that of methanol extract ranged from 0.5 to 10 mg/ml. The minimum bactericidal concentration (MBC) for ethanol extract ranged between 2.0 and 12.50 mg/ml, while this value for methanol ranged from 2.0 to 20 mg/ml. All the extracts exhibited appreciable activity against Candida albicans. The zones of inhibition exhibited by the extracts against C. albicans ranged between 15 and 18, 15 and 20 and 5 and 10 mm for ethanol, methanol and water extracts, respectively. Primary phytochemical screening revealed the presence of saponin, steroids, tannins, glycosides, alkaloids and flavonoids in the extracts. The ability of the crude stem extracts of O. basilicum to inhibit the growth of bacteria and fungi is an indication of its broad spectrum antimicrobial potential which may be employed in the management of microbial infections. It is also concluded that O. basilicum stem bark could be a potential source of active antimicrobial agents, and future works will be concentrated on the in vivo potencies and toxicological profiles.

The aim of this study was to determine the resistance rates of Helicobacter pylori (H. pylori) to various antimicrobial agents. The agar disk diffusion method (Kirby Bauer) was used to determine the sensitivity of H. pylori isolates to various antimicrobials. Of the 61 H. pylori isolates tested, no isolates was resistant to amoxycillin and tetracycline. The resistance rates were 42.6% for metronidazole, 21.3% for clarithromycin, and 3.3% for levofloxacin. Compared to clarithromycin and metronidazole, levofloxacin showed the lowest resistance. This is the first report on the resistance rates of H. pylori to antibiotics in Elazig Province, East of Turkey. This study suggests that the large scale studies is needed to help us to understand better the effect of resistance on the H. pylori eradication.

This study aimed to investigate the frequency of Varicella Zoster Virus (VZV) in benign and malignant breast tumors. A total of 24 carcinomas and 24 fibroadenomas paraffin embedded tumoral tissue samples were obtained from the pathology sections of Toos and Firoozgar hospitals in Tehran, Iran. All samples were collected from patients from June 2011 to February 2012. DNA was extracted from all samples and the infection with VZV was examined by PCR technique. The results obtained in this study showed that 3 out of 24 carcinoma samples (12.5%) were infected by VZV, while the number of fibroadenoma samples infected by this virus was 3 (12.5%). Based on the results obtained by this study, VZV was observed in both benign and malignant tumors. However, no association was observed between VZV infection and the formation of malignant or benign tumors based on the Chi-square test. Although it has been proven that VZV plays role in occurrence of latent infections after chemotherapy, no relationship was confirmed between this virus and breast benign and malignant tumors. Future studies are needed to elucidate the possible role of the virus in the disease. It is concluded that VZV is detectable in both malignant and benign tumors of the breast and the virus may play a role in the pathophysiology of breast tumors. However, further studies are needed to confirm the role of VZV in tumor formation.

According to the increased fungi contaminations and related damages, microbiologist's incentive in considering the fungal contaminations in human habitats had increased. Some of fungi cause disease through production of toxins in animals as well as humans. Since these toxins are not easily distinguishable, then it is crucial to study their characteristics. Aspergillus are among the most important toxigenic fungi which are found abundantly in Northern Iran habitats where is one of most important habitats and the main source of many feed and food stuffs in the state. Thus, we have decided to study the Deoxynivalenol (DON) production characteristics in culture medium by collection of Aspegillus isolates in Northern Iran. Samples were collected from Northern Alborz and southern Caspian Sea agricultural plants culturing and processing centers. The mould samples were isolated and identified based on CBS environmental sampling rules and ICPA diagnostics standards. They were then cultured to stimulate the toxin production till the targeted toxin to be measured at culturing substrate and fungi biomass. Afterward, they were exposed to extraction and then DON was measured by enzyme linked immunosorbent assay (ELISA). Amongst studied species, A. melleus (subgenus Circumdati/82.581 ppb) had the highest DON toxin production followed by A. spp. VI (subgenus Unclassifiable/50.803 ppb) and A. parasiticus (subgenus Circumdati/49.108 ppb). This report gives important information about the presence of DON in foodstuff. DON presence is a threat in Feed-Food processing; storing and packing which may be controllable by regular test and measuring of DON.