Hospital-Related Lineage of USA300 Methicillin-Resistant Staphylococcus aureus (MRSA) to Cause Bacteremia in Iran.

Staphylococcus aureus is an important pathogen that causes bloodstream infections. This study is aimed at assessing the genotypic characteristics of S. aureus strains responsible for bloodstream infections. An epidemiological study was conducted using 85 S. aureus strains isolated from bloodstream infections. Susceptibility was tested using the broth microdilution method and disk diffusion. All detected methicillin-resistant S. aureus (MRSA) isolates were confirmed by mecA PCR assays. S. aureus isolated from bacteremia were characterized using SCCmec, spa, and multilocus sequence typing methods. The prevalence of S. aureus strains responsible for bloodstream infections was 38.8%. All isolates were MRSA. Multidrug resistance (MDR) was present in 84.7% of isolates. MRSA isolated categorized within six clonal complexes including CC8 (60%), CC22 (22.4%), CC5 (5.9%), CC30 (4.7%), CC45 (4.7%), and CC59 (2.3%). The main lineages found were USA300/CC8-MRSA-IV/t008 (41.2%), followed by ST22-SCCmecIV/t790 (9.4%), ST239-SCCmecIII/t037 (7.1%), ST22-SCCmecIV/t032 (7.1%), ST239-SCCmecIII/t631 (5.9%), ST239-SCCmecIII/t860 (5.9%), ST22-SCCmecIV/t852 (5.9%), ST5-SCCmecIV/t002 (4.7%), ST45-SCCmecIV/t038 (4.7%), ST30-SCCmecIV/t318 (4.7%), ST59-SCCmecIV/t437 (2.3%), and ST225-SCCmecII/t045 (1.1%). Resistance to vancomycin amounted to 5.9% of isolates that belonged to ST239-SCCmecIII/t037 (80%) and ST8-SCCmecIV/t008 (20%). The emergence of USA300 strains in bloodstream infections in our country is a serious alarm and highlights the significant invasion of this lineage into the healthcare system. MDR patterns among these strains appear to be becoming the biggest problem in healthcare treatment.

The clinical outcomes of gut-brain axis (GBA) microbiota influence on psychiatric disorders.

The gut microbiome plays an important role in the health of the body. The study of its effect on mental problems has become the main topic of this study. As a matter of fact, every change in the gut microbiota composition can influence on mood and anxiety and vice versa. So, considering this "microbiota-gut-brain" axis (GBA) is so important. In this narrative review, the most recent reproduced information on GBA roles in neuropsychiatric disorders, and clinical significance have been considered. The gut microbial population is formed from birth and transforms from an immature state to the postnatal period into a more intricate and diverse adult ecosystem. In this review, we had some findings that GBA implicated in some psychiatric problems which can be a dysregulation consequence. In addition, some bacteria have been implicated in causing mental disorders in humans such as depression, obsessive-compulsive disorder, psychiatric disorders, stress disorders, schizophrenia and, autism. The absence of balance in GBA natural state can cause several negative consequences on host health which leads to neurological problems. Possibly, findings were delineating an interesting new etiological pathway for future exploration.

Antibiotic susceptibility of human-associated Staphylococcus aureus and its relation to agr typing, virulence genes, and biofilm formation.

BACKGROUND AND OBJECTIVE: Carriage of virulence factors confers some evolutionary benefit to bacteria, which favors the resistant strains. We aimed to analyze whether antibiotic susceptibility of Staphylococcus aureus strains is affected by agr typing, biofilm formation ability, and virulence profiles. METHODS: A total of 123 S. aureus clinical isolates were subjected to antimicrobial susceptibility testing by disk diffusion method, biofilm formation by microtiter plate method, as well as polymerase chain reaction screening to identify virulence genes and the accessory gene regulator (agr) types I-IV. A P value < 0.05 was considered significant. RESULTS: The most prevalent virulence gene was staphyloxanthin crtN, followed by hemolysin genes, capsular cap8H, toxic shock toxin tst, and enterotoxin sea, respectively. Resistant isolates were more commonly found in the agr-negative group than in the agr-positive group. Isolates of agr type III were more virulent than agr I isolates. Strong biofilm producers showed more antibiotic susceptibility and carried more virulence genes than non-strong biofilm producers. Associations were found between the presence of virulence genes and susceptibility to antibiotics. Carriage of the virulence genes and agr was higher in the inpatients; while, resistance and strong biofilms were more prevalent in the outpatients. CONCLUSION: These findings indicated the presence of several virulence factors, biofilm production capacity, agr types and resistance to antibiotics in clinical S. aureus isolates. Considering the importance of S. aureus for human medicine, an understanding of virulence and resistance relationships would help to reduce the impact of S. aureus infections.

Effect of Recombinant Helicobacter Outer Membrane Protein H (HopH) on Nitric Oxide Production by Peripheral Macrophage in BALB/c Mice.

BACKGROUND: Some products of bacteria are reported as an immunomodulator. The Helicobacter pylori (H. pylori) outer membrane proteins play an important role in stimulation of immune system. The present study was performed to determine the in vitro effect of recombinant HopH of H. pylori on Nitric Oxide (NO) production and viability of mouse peritoneal macrophages. METHODS: H. pylori recombinant HopH was produced in this study. Mice peritoneal macrophages were purified and cultured. Different concentrations of recombinant HopH were used for stimulation of macrophages in order to evaluate NO production. The cell viability was detected by MTT assay. NO amounts released in to the supernatants of cultured macrophages and LPS-stimulated macrophages (10 µg/ml) were detected by Griess reagent. RESULTS: Results demonstrated that the suppressive effect of high concentrations of recombinant HopH on NO release and the stimulation effect of protein was shown in 15 µg/ml, compared to the control group. NO stimulation was significant in all the concentrations of LPS stimulated with HopH groups. CONCLUSION: According to our findings, recombinant HopH has a toxic effect in high concentration on cell. So it can be an anticancer candidate.

Genetic Variability of Methicillin Resistant Staphylococcus Aureus Strains Isolated from Burns Patients.

OBJECTIVES: Staphylococcus aureus is a nosocomial pathogen that provides a major challenge in the healthcare environment, especially in burns units where patients are particularly susceptible to infections. In this study, we sought to determine molecular types of S. aureus isolates collected from burns patients, based on staphylococcal protein A and coagulase gene polymorphisms. METHODS: Antibiotic susceptibility testing of 89 S. aureus strains isolated from burn wounds of patients was assessed using the Kirby-Bauer disk diffusion method. Strains were characterized by spa typing, coa typing, and resistance and toxin gene profiling. RESULTS: A total of 12 different spa types were identified with the majority being t790 (18%). Panton-Valentine leucocidin encoding genes were identified in spa types t044 (5.6%), t852 (2.2%) and t008 (2.2%). The most commonly detected antibiotic resistance gene was ant (4')-Ia (60.7%). Ten different coa types were detected and the majority of the tested isolates belonged to coa III (47.2%). All the high-level mupirocin-resistant and low-level mupirocin resistant strains belonged to coa type III. CONCLUSION: The present study illustrated that despite the high frequency of coa III and spa t790 types, the genetic background of S. aureus strains in Iranian burns patients was diverse. The findings obtained are valuable in creating awareness of S. aureus infections within burns units.

Phenotypic and Molecular Characterization of Methicillin-Resistant Staphylococcus aureus Clones Carrying the Panton-Valentine Leukocidin Genes Disseminating in Iranian Hospitals.

The spread of Panton-Valentine leukocidin (PVL)-carrying Staphylococcus aureus strains in both hospital and the community is a significant worldwide problem. The aim of the study was to investigate the clonal dissemination pattern of PVL-producing S. aureus strains isolated from hospitalized patients in Tehran, Iran. In this cross-sectional study, 70 PVL-carrying S. aureus strains were recovered from 240 clinical specimens and characterized by antibiotic susceptibility testing, agr typing, SCCmec typing, spa typing, multilocus sequence typing, and virulence and adhesion gene profiling. All the PVL-carrying S. aureus strains were confirmed as methicillin-resistant S. aureus (MRSA) and recovered from wounds (48.6%), blood (25.7%), exudate/pus (11.4%), sputum (8.6%), and body fluid (5.7%) samples. Among the 70 PVL-carrying S. aureus strains tested, 38 (54.3%) were positive for ant (4')-Ia gene, 27 (38.6%) for aac (6')-Ie/aph (2″), 13 (18.6%) for msr(A), 13 (18.6%) for erm(C), 13 (18.6%) for tet(M), 11 (15.7%) for erm(A), 10(14.3%) for msr(B), 9 (12.9%) for aph (3')-IIIa, 5 (7.1%) for mupA, and 2 (2.9%) for erm(B) genes. Five clonal complexes (CC) and nine different clones were detected in this study. The most frequent CC was CC22 (ST22) (42.8%) followed by CC30 (ST30) (21.5%), CC8 (ST8) (17.2%), CC1 (ST772) (11.4%), and CC80 (ST80) (7.1%). In this study, ST22-SCCmec IV/t852 was the predominant PVL-positive MRSA clone (20%), followed by ST8-SCCmec IV/t008 (17.2%), ST30-SCCmec IV/t019 (12.9%), ST22-SCCmec IV/t790 (11.4%), ST22-SCCmec IV/t005 (11.4%), ST30-SCCmec IV/t021 (8.6%), ST80-SCCmec IV/t044 (7.1%), ST772-SCCmec V/t657 (7.1%), and ST772-SCCmec V/t10795 (4.3%). Diversity in clonal types of PVL-carrying MRSA strains in our study supports the need to perform a systematic surveillance of PVL-positive MRSA strains.

Identification of a Novel Cassette Array in Integronbearing Helicobacter Pylori Strains Isolated from Iranian Patients.

Helicobacter pylori as the second most common cause of gastric cancer in the world infects approximately half of the developed countries population and 80% of the population living in developing countries. Integrons as genetic reservoirs play major roles in dissemination of antimicrobial resistance genes. To the best of our knowledge, this is the first study to report carriage of class 1 and 2 integrons and associated gene cassettes in H. pylori isolates from Iran. This crosssectional study was conducted in Tehran among 110 patients with H. pylori infection. Antimicrobial susceptibility testing (AST) for H. pylori strains were assessed by the micro broth dilution method. Class 1 and 2 integrons were detected using PCR. In order to determine gene cassettes, amplified fragments were subjected to DNA sequencing of both amplicon strands. The prevalence of resistance to clarithromycin, metronidazole, clarithromycin, tetracycline, amoxicillin, rifampin, and levofloxacin were 68.2% (n=75), 25.5% (n=28), 24.5% (n=27), 19.1% (n=21), 18.2% (n=20) and 16.4% (n=18), respectively. Frequency of multidrug resistance among H. pylori isolates was 12.7%. Class 2 integron was detected in 50 (45.5%) and class 1 integron in 10 (9.1%) H. pylori isolates. The most predominant gene cassette arrays in class 2 integron bearing H. pylori were included sateraaadA1, dfrA1sat2aadA1, blaoxa2 and, aadB whereas common gene cassette arrays in class 1 integron were aadBaadA1cmlA6, aacA4, blaoxa2, and catB3. The high frequency of class 2 integron and multidrug resistance in the present study should be considered as a warning for clinicians that continuous surveillance is necessary to prevent the further spread of resistant isolates.

Phylogenetic grouping and pathotypic comparison of urine and fecal Escherichia coli isolates from children with urinary tract infection.

The aim of this study was to investigate the phylogenetic background and to assess hlyD (involved in the secretion of haemolysin A) and intI1 (encoding a class 1 integrase) in Escherichia coli isolates derived from urinary and fecal specimens. A total of 200 E. coli isolates was collected from patients presenting with urinary tract infection (UTI) during September 2009 to September 2010 and screened for hlyD and intI1 genes by polymerase chain reaction (PCR). Phylogenetic analysis showed that E. coli is composed of four main phylogenetic groups (A, B1, B2 and D) and that uropathogenic E. coli (UPEC) isolates mainly belong to groups B2 (54%) and D (34%) whereas group A (44%) and D (26%) are predominant among commensal E. coli isolates. In this study, hlyD was present in 26% of UPEC and 2% of commensal E. coli isolates. However, hemolytic activity was detected for 42% of UPEC and 6% of commensal E. coli isolates (p < 0.05). intI1 gene was more frequently expressed in UPEC (24%) in comparison with commensal E. coli isolates (12%). Resistance to aztreonam, co-trimoxazole and cefpodoxime were frequently found among UPEC isolates whereas commensal E. coli isolates were commonly resistant to co-trimoxazole, nalidixic acid and cefotaxime. Concluding, a considerable difference between UPEC and commensal E. coli isolates was observed regarding their phylogenetic groups, presence of class 1 integron and hlyD gene, hemolysin activity and resistance pattern. The detection of class 1 integrons and hlyD gene was higher among UPEC compared with commensal E. coli isolates. These findings may contribute for a better understanding of the factors involved in the pathogenesis of UPEC.

Phylogenetic grouping and pathotypic comparison of urine and fecal Escherichia coli isolates from children with urinary tract infection

The aim of this study was to investigate the phylogenetic background and to assess hlyD (involved in the secretion of haemolysin A) and intll (encoding a class 1 integrase) in Escherichia coli isolates derived from urinary and fecal specimens. A total of 200 E. coli isolates was collected from patients presenting with urinary tract infection (UTI) during September 2009 to September 2010 and screened for hlyD and intll genes by polymerase chain reaction (PCR). Phylogenetic analysis showed that E. coli is composed of four main phylogenetic groups (A, B1, B2 and D) and that uropathogenic E. coli (UPEC) isolates mainly belong to groups B2 (54%) and D (34%) whereas group A (44%) and D (26%) are predominant among commensal E. coli isolates. In this study, hlyD was present in 26% of UPEC and 2% of commensal E. coli isolates. However, hemolytic activity was detected for 42% of UPEC and 6% of commensal E. coli isolates (p < 0.05). intll gene was more frequently expressed in UPEC (24%) in comparison with commensal E. coli isolates (12%). Resistance to aztreonam, co-trimoxazole and cefpodoxime were frequently found among UPEC isolates whereas commensal E. coli isolates were commonly resistant to co-trimoxazole, nalidixic acid and cefotaxime. Concluding, a considerable difference between UPEC and commensal E. coli isolates was observed regarding their phylogenetic groups, presence of class 1 integron and hlyD gene, hemolysin activity and resistance pattern. The detection of class 1 integrons and hlyD gene was higher among UPEC compared with commensal E. coli isolates. These findings may contribute for a better understanding of the factors involved in the pathogenesis of UPEC.

Determination of integron frequency by a polymerase chain reaction-restriction fragment length polymorphism method in multidrug-resistant Escherichia coli, which causes urinary tract infections.

The purpose of this study was to determine the presence of integrons in Escherichia coli, which cause urinary tract infections, and to define the association between integrons and antimicrobial susceptibility. Susceptibility of 200 isolates from urine samples of patients suffering from urinary tract infections to 13 antibiotics was determined by the Kirby-Bauer disk diffusion method. The existence of class1 and 2 integrons in resistant isolates was assessed by polymerase chain reaction-restriction fragment length polymorphism and sequencing. Antibiotic resistance patterns were observed as follows: amoxicillin 78%, tetracycline 76.1%, co-trimoxazole 67.7%, cephalotin 60%, nalidixic acid 57.4%, chloramphenicol 49%, gentamicin 46.4%, ceftazidim 38.1%, ciprofloxacin 36.2%, nitrofurantoin 33.5%, amikacin 32.1%, norfloxacin 36.1%, and imipenem 27.1%. Of 200 isolates, 155 (77.5%) were multidrug resistant (MDR). The existence of integrons was confirmed in 50.3% of isolates. Three class 1 integron types, aadA2 being the most frequently found, and four class 2 integron types are described. Significant association between resistance to gentamicin, co-trimoxazole, cephalotin, ceftazidim, imipenem, chloramphenicol, and nalidixic acid with the existence of integrons was observed. Multidrug resistance suggests that the strategy for treatment of patients with E.coli infections needs to be revised. Furthermore, it was shown that integrons may be partly responsible for multidrug resistance. Imipenem and norfloxacin were the most effective antibiotics against isolates.

Study Prevalence of Verotoxigenic E.coli Isolated from Urinary Tract Infections (UTIs) in an Iranian Children Hospital.

BACKGROUND AND OBJECTIVES: Urinary tract infections (UTI) caused by enterohemorrhagic Escherichia coli (EHEC) is one of the most important diseases in infants and children. If there would not be any useful diagnosis and treatment it may be resulted in diseases such as acute renal failure, thrombocytopenia and hemolytic anemia. The aim of this study was to determine frequency of verotoxigenic E.coli isolates in urine of children with (UTIs) in Mofid children Hospital. METHODS: During one year from September 2008 to august 2009, urine specimens were taken from children who suspected to UTI admitted to Mofid Children Hospital. E.coli strains that indicated beta hemolytic on sheep blood agar, negative sorbitol fermentation on SMAC (sorbitol macconky agar) and negative motility on SIM were tested by PCR and serologic (VITEC-RPLA kit) methods for detecting toxin genes and production of toxin, respectively. RESULTS: Among 12572 urine specimens were taken from children admitted to Mofid hospital, we isolated 378 E.coli from urine samples which only 9 isolates were EHEC. Only five EHEC strains (55%) which produced vtx genes, were detected by serologic and PCR methods. CONCLUSION: The prevalence of urinary infections caused by EHEC strains is very significant because it causes aggravating pathologic effects. Thus we suggest rapid method for identification of this bacteria and proper treatment to Inhibition of unwanted complications.

Torque teno virus and hepatitis C virus co-infection in Iranian pediatric thalassemia patients.

OBJECTIVE: Torque teno virus (TTV) infects patients at risk for parenteral exposure and chronic blood transfusion, such as those with ß-thalassemic. This study aimed to assess the prevalence of TTV infection and co-infection of TTV and hepatitis C virus (HCV) in pediatric thalassemia patients receiving chronic blood transfusion. MATERIAL AND METHODS: The study included 90 pediatric thalassemia patients receiving chronic blood transfusion that presented to the Mofid Children's Hospital, Tehran, Iran. The control group included 90 healthy volunteer children. Serum TTV DNA detection via semi-nested PCR and HCV Ab were performed in all the participants. Demographic characteristics and clinical data were collected from each participant for statistical analysis. RESULTS: In all, 64.4% of the patients had TTV infection, versus 24.4% of the controls (P < 0.01). The thalassemia patients had a greater probability of having TTV and HCV infections than the controls, with a common OR of 5.60 (95% CI: 2.94-10.69) and 2.15 (95% CI: 1.83-2.50), respectively. In total, 17.2% (10/58) of the patients that were TTV positive were also HCV positive, whereas 6.3% (2/32) of the TTV-negative patients were anti-HCV antibody (Ab) positive (P = 0.14). CONCLUSION: The prevalence of TTV and HCV infection was higher in the Iranian thalassemia patients on chronic transfusion therapy than in the controls. The high prevalence of TTV in pediatric thalassemia patients on chromic transfusion therapy may indicate the superiority of the parenteral route compared to other routs of TTV transmission.

Clinical outcomes of Torque teno virus-infected thalassemic patients with and without hepatitis C virus infection.

BACKGROUND: Although a marked proportion of thalassemic patients acquire Torque teno virus (TTV) through blood transfusion, its clinical importance is unclear. This study was designed to investigate the clinical importance of TTV infection in thalassemic patients with and without hepatitis C virus (HCV) co-infection in Iran. METHODS: In this case-control study, 107 thalassemic patients on chronic transfusion and 107 healthy individuals were selected. According to HCV and TTV infection status (detected by semi-nested PCR), patients were categorized into 4 groups: TTV and HCV negative, TTV positive, HCV positive, and TTV and HCV positive. Blood ferritin, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels in these 4 groups were assessed. RESULTS: Approximately half of the thalassemic patients (50.5%) and 27.1% of controls had TTV infection. Thalassemic patients had a greater chance of TTV infection compared to the control group with a sex-adjusted OR of 4.13 (95% CI=2.28-8.13). The increased levels of ALT, AST, and ferritin in the TTV and HCV-infected group were not significantly different from those in the TTV and HCV negative group. Co-infection with TTV and HCV did not significantly increase ALT, AST, and ferritin levels compared to infection with TTV alone. CONCLUSION: Although common in thalassemic patients, TTV infection appears to have a negligible role in increasing the severity of liver disease, even when co-infection with HCV occurs.