EVALUATION OF NON H. PYLORI SPIRAL ORGANISMS IN HUMAN GASTRIC BIOPSIES BY USING PCR AND MICROSCOPIC METHODS IN IRAN (FIRST REPORT)

Introduction and Objectives: The Discovery of Helicobacter pylori in 1982 increased interest in the range of other spiral bacteria that had been seen in Stomach (Marshall & Warren 1984).The power of technologies such as the polymerase chain reaction (PCR) with genus specific primers revealed that many of these bacteria belong to the genus Helicobacter. These nonpylori helicobacters are increasingly being found in human clinical specimens. Non-pylori Helicobacters are Gram-negative, motile, long, tightly coiled, Spiral bacteria ,with three to eight coils, that cause of some gastric problems like gastritis, peptic ulceration and Mcosa-Associated Lymphoid Tissue (MALT) lynphoma in animals and humans. Materials and Methods: Samples taken during endoscopy were analyzed by rapid urease test, PCR and light microscope(Giemsa and Gram staining). In this study 270 samples were collected from Patients with gastric disorders. Presence of Helicobacters confirmed by a positive urease test and Helicobacter genus specific PCR method utilized. DNA was prepared from biopsies using the Qiamp tissue kit (QIAGEN Inc., Valencia, Calif.) and frozen at −20°C (like gastric samples/biopsies). DNA samples that PCR positive were amplified with 16SrRNA gene primers against Helicobacter species. Results: In gastric biopsy specimen's non-pylori helicobacter spp., have been observed. At the end of the study we found that 71% of urease tests, 0.37% of light microscopic studies (we observed some spiral gram negative bacteria with 2-7 coils) and 0.74% of PCR tests were positive. In analysis with PCR route 2 person (both of them were Male) were infected with H.heilmanniilike organisms( one of them kept a dog for 5 years as a pet).16S rRNA gene amplification was performed on 270 DNA samples and results were positive for H.heilmannii in two cases (275-bp), but negative for H.bizzozeronnii,H.felis and H. Salmonis

Comparison of culture and PCR methods for diagnosis of vaginal infection due to Mycoplasma Hominis

Background and Objective: Mycoplasma Hominis is the smallest pathogenic bacteria, with no cell wall and free living organisms. It grows slowly and the conventional clinical microbiology techniques can not be applied due to difficulties in cultivation in particular slow growth incubation. This study was done to compare the culture and PCR methods for diagnosis of vaginal infection due to Mycoplasma Hominis. Methods: This laboratory test evaluation study was done on 150 patients with bacterial vaginosis and 50 healthy people with no infection as control, whom refereed to Imam Khomeini and Imam Zaman Hospitals in Tehran. Samples were collected in PPLO culture for growth and PBS to perform PCR method. Results: 35.3% and 76% of patients were positive using culture and PCR methods, respectively. Using PCR method 8% of control subjects was positive. There was no significant association between PCR method with abortion, place of residence and also level of educations. There was a significant association between the age (P<0.05), times of changing under wear cloths (P<0.05) and parity (P<0.05). Conclusion: PCR method is a more reliable technique to detect the vaginal infection due to Mycoplasma Hominis compared to culturing

Chemical composition, antibacterial and radical scavenging activity of essential oils from Satureja macrantha C.A.Mey. at different growth stages

Essential oils (EOs) from medicinal and aromatic plants are interesting products to be used as natural food preservatives. The EOs from the genus Satureja are reported to inhibit foodborne pathogens being worthy of use as food preservatives. Satureja macrantha is found in Western and Northwest Iran and commonly used as a food flavoring agent and for the treatment of urinary diseases. The objective of the present study was to identify the chemical composition of S. macrantha EOs at different growth stages (vegetative, flowering and fruiting stages) and to evaluate their biological activities. Chemical compositions were analyzed using GC-FID and GC-MS. The antibacterial activity was evaluated using the broth microdilution method against the foodborne pathogenic bacteria Staphylococcus aureus (ATCC23922), Enterococcus faecalis (ATCC29212) (Gram-positive), Enterobacter aerogenes (ATCC13046) and Escherichia coli. The antioxidant activity was estimated using the DPPH, ABTS and reducing power assays. The yields of S. macrantha EOs were in the range of 1.4–1.8%, thus scalable for the manufacture of food preservatives on an industrial level. The main compounds of EOs were carvacrol (42.7–48.2%), thymol (0.2–16.5%), p-cymene (10.1–14.7%) and γ-terpinene (7.9–9.1%) in all phenological stages examined. The strongest antibacterial activity (MICs = 5–20 μg/mL) of the EOs was recorded in samples obtained during the flowering stage where carvacrol (42.7%) and thymol (16.5) were present both at high percentages. On the other hand, the antioxidant activity was found to be slightly higher in the other stages. As the EO obtained at flowering showed the best inhibitory properties against foodborne pathogenic bacteria, it is suggested that plants at this stage can be selected as main sources of food preservative agents

Non H.pylori Helicobacter Identified as H.heilmannii in Gastric Biopsy samples in humans with gasteric disorders by PCR and Microscopic Methods in IRAN (First Report)

The Discovery of Helicobacter pylori in 1982 increased interest in the range of other spiral bacteria that had been seen in Stomach.The power of technologies such as the polymerase chain reaction (PCR) with genus specific primers revealed that many of these bacteria belong to the genus Helicobacter. These non-pylori helicobacters are increasingly being found in human clinical specimens. Non-pylori Helicobacters are Gram-negative, motile, long, tightly coiled, Spiral bacteria ,with three to eight coils, that cause of some gastric problems like gastritis, peptic ulceration and Mcosa-Associated Lymphoid Tissue (MALT) lynphoma in animals and humans. Samples taken during endoscopy were analyzed by rapid urease test, PCR and light microscope(Giemsa and Gram staining). In this study 810 biopsy samples from 270 patients with gastric disorders were collected. Presence of Helicobacters confirmed by a positive urease test and Helicobacter genus specific PCR method utilized. DNA was prepared from biopsies using the Qiamp tissue kit (QIAGEN Inc., Valencia, Calif.) and frozen at -20°C (like gastric samples/biopsies). DNA samples were amplified with 16SrRNA gene primers against Helicobacter species. In gastric biopsy specimen's non-pylori helicobacter spp., have been observed. At the end of the study we found that 71% of urease tests, 0.37% of light microscopic studies (we observed some spiral gram negative bacteria with 2-7 coils) and 0.74% of PCR tests were positive. In analysis with PCR route 2 person (both of them were Male) were infected with H.heilmannii-like organisms( one of them kept a dog for 5 years as a pet).16S rRNA gene amplification was performed on 270 DNA samples and results were positive for H.heilmannii in two cases (275-bp), but negative for H.bizzozeronnii,H.felis and H. Salmoni

Phenotypic and Genotypic Assay for Detection of Extended Spectrum beta-lactamases Production by Klebsiella pneumoniae Isolates in Emam Reza Hospital in Tabriz, Iran

Objectives of this study were to investigate the prevalence of K. pneumoniae producing ESBLs, to evaluate the susceptibility of K. pneumoniae producing ESBLs towards non-beta-lactam antibiotics and to study the dominant ESBLs gene in Emam Reza hospital. K. pneumoniae producing ESBLs identified by phenotypic and genotypic methods. Polymerase Chain Reaction (PCR) performed for detection of blaSHV, TEM and CTX-M. The findings showed that 43.69, 13.59, 7.77, 11.65 and 23.3 were from UT!, ICUs, surgery ward, lesion infections and RTI, respectively. The results showed that 43.7 of isolates were ESBLs produces. The findings revealed that 26.7, 6.7, 20 and 0 of K.pneumoniae producing ESBLs were resistant to amikacin, ciprofloxacin, cotrimoxazol and imipenem, respectively. Thirty-nine blaSHV, seven blaTEM and seven blaCTX-M identified among K.pneumoniae producing ESBLs. The results reflected in cold month resistant to third generation cephalosporins were more than warm months. Generally, frequency of blaSHV was more than blaCTX-M and blaTEM