Genetic Diversity of Helicobacter pylori Isolates from Patients with Gastric Diseases in Isfahan.

Background: Helicobacter pylori (H. pylori), a spiral-shaped bacterium colonizing the human stomach, is generally acquired in childhood. This pathogen is highly diverse and can be used as genetic markers for predict the history of human migrations. This study aimed to determine the genetic diversity of H. pylori isolates from patients with dyspepsia by the multi-locus sequence typing (MLST) and update data on the prevalence of H. pylori among Iranian dyspeptic patients. Materials and Methods: In this descriptive cross-sectional study, 165 gastric biopsy specimens were obtained from patients with dyspepsia referred to Dr. Shariati Hospital of Isfahan, Iran, from April to July 2018. The status of H. pylori infection was determined by FISH in paraffin-embedded biopsy specimens. MLST of seven housekeeping genes was performed for 20 H. pylori isolates. The phylogenetic tree was plotted using CLC v8 and iTol software. Results: The overall prevalence of H. pylori infection was 53.3%. In the results of the analysis of MLST, a total of 14 new STs were recorded. The results of the global analysis showed that all the isolates, with a wide diversity, have a genetic affinity with members of the European population, such as Italy and Russia, and are in the hpEurope haplotype. Conclusion: Given the high prevalence of H. pylori infection in this region, early and accurate identification of patients seems necessary. Sequence analysis and determination of the origin of the phylogeny of strains can be effective in clinical management and monitoring of risk factors for chronic and recurrence of infection.

The characteristic and potential therapeutic effect of isolated multidrug-resistant Acinetobacter baumannii lytic phage.

BACKGROUND: Widespread misuse of antibiotics caused bacterial resistance increasingly become a serious threat. Bacteriophage therapy promises alternative treatment strategies for combatting drug-resistant bacterial infections. In this study, we isolated and characterized a novel, potent lytic bacteriophage against multi-drug resistant (MDR) Acinetobacter baumannii and described the lytic capability and endolysin activity of the phage to evaluate the potential in phage therapy. METHODS: A novel phage, pIsf-AB02, was isolated from hospital sewage. The morphological analysis, its host range, growth characteristics, stability under various conditions, genomic restriction pattern were systematically investigated. The protein pattern of the phage was analyzed, and the endolysin activity of the phage was determined under the non-denaturing condition on SDS-PAGE. The optimal lytic titer of phage was assessed by co-culture of the phage with clinical MDR A. baumannii isolates. Finally, HeLa cells were used to examine the safety of the phage. RESULTS: The morphological analysis revealed that the pIsf-AB02 phage displays morphology resembling the Myoviridae family. It can quickly destroy 56.3% (27/48) of clinical MDR A. baumannii isolates. This virulent phage could decrease the bacterial host cells (from 108 CFU/ml to 103 CFU/ml) in 30 min. The optimum stability of the phage was observed at 37 °C. pH 7 is the most suitable condition to maintain phage stability. The 15 kDa protein encoded by pIsf-AB02 was detected to have endolysin activity. pIsf-AB02 did not show cytotoxicity to HeLa cells, and it can save HeLa cells from A. baumannii infection. CONCLUSION: In this study, we isolated a novel lytic MDR A. baumannii bacteriophage, pIsf-AB02. This phage showed suitable stability at different temperatures and pHs, and demonstrated potent in vitro endolysin activity. pIsf-AB02 may be a good candidate as a therapeutic agent to control nosocomial infections caused by MDR A. baumannii.

Molecular Assessment of Resistance to Clarithromycin in Helicobacter pylori Strains Isolated from Patients with Dyspepsia by Fluorescent In Situ Hybridization in the Center of Iran.

BACKGROUND AND AIMS: Helicobacter pylori is a common infectious bacterium mostly found in gastroduodenal diseases. The increased prevalence of clarithromycin-resistant H. pylori strains is a major challenge in the successful treatment of infections caused by this organism. The present study is aimed at detecting the clarithromycin resistance pattern of H. pylori strains isolated from gastric biopsies and evaluating point mutations of the 23S rRNA gene. Patients and methods. In the present descriptive cross-sectional study, 165 patients with gastrointestinal disorders, who were referred to the Endoscopy Center of Dr. Shariati Hospital of Isfahan, Iran, were enrolled from April to July 2018. H. pylori infection was diagnosed by culture, and susceptibility of the isolates to clarithromycin was assessed by the E-test. Minimum inhibitory concentration (MIC) values were obtained based on EUCAST recommendations. Also, fluorescence in situ hybridization (FISH) was used to determine point mutations associated with clarithromycin resistance. RESULTS: By using culturing, H. pylori was isolated from 50.3% (83/165) gastric biopsy specimens. The overall frequency of resistance to clarithromycin was 25.3% (21/83) by the E-test. In the resistance genotypic analysis, 19 isolates had mutations. The prevalence of A2143G and A2144G mutations was 68.4% (13/19) and 31.5% (6/19), respectively. A2143C mutation was not tracked in any isolate. Two isolates with MIC > 0.5 µg/mL had no mutations that could be related to other mechanisms of resistance. CONCLUSION: As presented in the study, the high prevalence of clarithromycin-resistant H. pylori due to point mutations of the 23S rRNA gene indicates the necessity of revising the standard treatment regimen based on antibiotic susceptibility pattern of each region.

Evaluation of biofilm removal activity of Quercus infectoria galls against Streptococcus mutans.

BACKGROUND: Dental caries is one of the most prevalent infectious diseases affecting humans of all ages. Streptococcus mutans has an important role in the development of dental caries by acid production. The purpose of this study was to evaluate the antibacterial and biofilm disinfective effects of the oak tree Quercus infectoria galls against S. mutans. MATERIALS AND METHODS: The bacterial strain used in this study was S. mutans (ATCC: 35668). Two kinds of galls, Mazouj and Ghalghaf were examined. Galls were extracted by methanol, ethanol and acetone by Soxhlet apparatus, separately. Extracts were dissolved in sterile distilled water to a final concentration of 10.00, 5.00, 2.50, 1.25, 0.63, 0.31, and 0.16 mg/ml. Microdilution determined antibacterial activities. The biofilm removal activities of the extracts were examined using crystal violet-stained microtiter plate method. One-way ANOVA was used to compare biofilm formation in the presence or absence of the extracts. RESULTS: The methanolic, ethanolic, and acetonic extracts of Q. infectoria galls showed the strong inhibitory effects on S. mutans (P < 0.05). The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values for the Mazouj and Ghalghaf gall extracts against S. mutans were identical. The MIC values ranged from 160 µg/ml to 320 µg/ml, whereas the MBC values ranged from 320 µg/ml to 640 µg/ml. All extracts of Q. infectoria galls significantly (P < 0.05) reduced biofilm biomass of S. mutans at the concentrations higher than 9.8 µg/ml. CONCLUSION: Three different extracts of Q. infectoria galls were similar in their antibacterial activity against S. mutans. These extracts had the highest biofilm removal activities at 312.5 µg/ml concentration. The galls of Q. infectoria are potentially good sources of antibacterial and biofilm disinfection agent.