Comparison of the Prevalence of Microbial Surface Components Recognizing Adhesive Matrix Molecules (MSCRAMMs) among Staphylococcus aureus Isolates in a Burn Unit with Non-Burning Units.

BACKGROUND: Staphylococcus aureus (S. aureus) is one of the most important pathogens in burn infections colonized in the nose and increase the risk of infections. METHODS: Overall, 85 S. aureus isolates were isolated from clinical and nasal hospitalized patients and health care workers (HCWs) in a burn unit and non-burn units in Isfahan from June 2016 and September 2016. Genes encoding penicillin-binding protein 2a (mecA) and adhesive surface proteins, including fibronectin-binding proteins (fnbA,fnbB), fibrinogen binding protein (fib), laminin-binding protein(eno), collagen binding protein (cna), elastin binding protein (ebps), intracellular adhesion operon (icaA and icaD) were detected using PCR method. RESULTS: The rate of methicillin-resistant S. aureus (MRSA) among burn and non-burn isolates were 62% (18/29) and 25% (14/56), respectively. The most prevalent MSCRAMMs genes in burn units were eno (86%) and fib (66%). The most common gene pattern in burn center was icaA+fib+eno. The frequency of icaD, fib and ebpS was higher in clinical samples than nasal samples. No relation was found between the MSCRAMMs genes in the burn unit and non-burn units. CONCLUSION: The high prevalence of MRSA in burn center can be a new challenge for clinicians. The higher frequency of icaD, fib and ebpS in clinical isolates than nasal isolates may reflect the important role of these genes in colonization and pathogenesis of S. aureus.

Molecular Typing of Community-acquired Methicillin-Resistant Staphylococcus aureus Isolated from 2- to 6-year old Children by Staphylococcal Protein A and Agr Typing in Isfahan, Iran.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) has become a considerable public health concern in the entire world due to the rapid spread of this bacterium in human community; also the epidemiology of MRSA has changed, as the isolation of MRSA strains from healthy and non-healthy patients. Therefore, the objective of this study is to determine the genetic diversity and antibiotic resistance profile of community-acquired (CA)-MRSA nasal carriage in the Iranian samples. MATERIALS AND METHODS: A total of 25 CA-MRSA were isolated from the anterior nares of 410 healthy preschool children. All MRSA isolates were characterized by the detection of the toxic shock syndrome toxin-1 (TSST-1) and typed by γ-hemolysin genes, agr groups, and staphylococcal protein A (spa) typing. Kirby-Buyer antibiotic susceptibility testing was performed and interpreted as per the standard guidelines. RESULTS: A total of 25 (6.1%) MRSA isolates were recovered from the anterior nares of 410 preschool children. Sixteen isolates (64%) were positive for the TSST-1 gene. Three agr specificity groups were determined, as follows: eight (32%) isolates belonged to agr Group I, five (20%) isolates belonged to agr Group II, and 12 (48%) isolates belonged to agr Group III. The repeated profiles of these spa types of 25 isolates were organized into eight different lineages groups. Five of lineages contained a single strain, three of lineages contained two strains, and three of lineages consisted of more than three strains. CONCLUSIONS: The results of our study show that the rate of MRSA in our region is significantly high. Additionally, spa type t037 was the predominant type among CA S. aureus.

Antimicrobial Resistance in Hospitalized Patients with Community Acquired Urinary Tract Infection in Isfahan, Iran.

BACKGROUND: Antibiotic resistance against uro-pathogens is a worldwide health concern. The aim of this study was to determine the causative bacteria and antibiotic susceptibility patterns among hospitalized patients with community acquired urinary tract infection (UTI). METHODS: This cross-sectional study was performed in 2016-2018 in Isfahan, Iran. Urine samples were examined for strain identification and antimicrobial resistance pattern using standard tests. Stratification was done based on gender and age (<20 and >20 years) groups. Chi-square and Fisher exact tests were applied to assess differences in etiology and susceptibility rates between groups. RESULTS: Among 1180 patients, Escherichia coli was the commonest pathogen (68.1%) followed by Enterococcus spp. (8.8%) and Klebsiella pneumonia (8.0 %). Non-E. coli pathogens were more frequent among males (41.8% versus 24.8% in females, P<0.01) and in those aged under 20 years (61.0% versus 22.2% in older than 20 years, P<0.01). Isolated bacteria revealed high susceptibility to imipenem (94.9%), meropenem (92.2%), and amikacin (91.9%); moderate sensitivity to gentamicin (64.4%), cefepime (52.6%) and ceftazidime (47.2%); and low susceptibility to ceftriaxone (41.8%), cefotaxime (40.0%), ciprofloxacin (38.6%) and trimethoprim-sulfamethoxazol (31.3%). The sensitivity of isolates to ceftriaxone, ceftazidime, cefepime, imipenem, meropenem, amikacin and ciprofloxacin was significantly higher in females. Compared to the older age group, uro-pathogens were more susceptible to ciprofloxacin, ceftazidime and gentamicin in patients aged under 20 years. CONCLUSION: We found that imipenem, meropenem and amikacin were good choices for empiric therapy of complicated or severe hospitalized patients with community acquired UTI; and gentamicin, cefepime and ceftazidime were acceptable as initial choices in non-severe infections in the area.

Methodology and Early Results of the First Surveillance Program on Prevention and Control of Antimicrobial Resistance in Isfahan, Iran: The IAS-I Study.

BACKGROUND: Isfahan Antibiotic Resistance Surveillance System-1 has been instituted in Isfahan, Iran to construct a project for surveillance of clinically significant bacteria, and to help raise a logic regional stewardship program for prevention and control of disseminating-resistant organisms. METHODS: During March 2016 to March 2018, an antibiotic resistance surveillance system was designed and implemented by Isfahan Infectious Diseases and Tropical Medicine Research Center. The surveillance program was implemented in three general hospitals in Isfahan. In addition to the routine microbiology data, clinical data (differentiation between true infections and contamination, healthcare-associated infections (HCAI) and community-acquired infections (CAI), as well as determination of the infection site) were obtained and analyzed by WHONET software. RESULTS: During a 2-year period, from 7056 samples that revealed growth of bacteria, 3632 (51.5%) isolates were detected as contamination and 3424 (48.5%) true bacterial isolates were identified. Of these, about 32% of isolates were recognized as HCAI. Totally, the most recognized infections were urinary tract infection, bloodstream infection and skin and soft tissue infections. In patients with HCAIs, 70% of isolates were gram negative and in patients with CAIs 73% isolates were gram negative bacteria. CONCLUSIONS: The strength of the project is gathering enough clinical information in addition to microbiologic data, which would increase application of the results for empiric treatment and prevention of the infectious diseases in clinical settings.

Molecular characterization and genotyping of methicillin-resistant Staphylococcus aureus in nasal carriage of healthy Iranian children.

PURPOSE: Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has become a considerable public health concern in both developed and developing countries due to the rapid spread of this bacterium around the world, also the epidemiology of MRSA has changed, as the isolation of MRSA strains is not limited to health-care settings or patients with predisposing risk factors. Therefore, the objective of this study is to determine the genetic diversity and antibiotic resistance profile of CA-MRSA nasal carriage in Iranian children. METHODOLOGY: A cross-sectional study was conducted from April 2013 to March 2014. A total of 25 CA-MRSA were isolated from the anterior nares of 410 preschool children with no risk factors. All MRSA isolates were characterized by detection of the Panton-Valentine leukocidin (pvl) and γ-hemolysin genes, staphylococcal cassette chromosome mec (SCCmec) typing and multi-locus sequence typing (MLST). RESULTS: In 25 CA-MRSA isolates, Pvl and γ-hemolysin genes were detected in one (4%) and 18 (72 %) isolates; respectively. Overall, 92% (23/25) of isolates belonged to SCCmec type IV and 8% (2/25) of them had SCCmec type V profile. Using MLST, the 25 isolates were grouped into six clonal complexes (CC) and eight sequence types (ST) (CC5/ST6, CC22/ST22 and ST217, CC30/ST30 and ST1107, CC78/ST859, CC398/ST291 and CC97/ST405). The ST859/SCCmec IV (11/25, 44%) was the predominant clone among the isolates. ST859-MRSA-IV-pvl-negative (resistant to tetracycline) have successfully adapted to the Iranian preschool children population. CONCLUSION: Our results suggest that the genomic diversity was observed among the CA-MRSA. In addition, the current study demonstrates that pvl is not a reliable marker for CA-MRSA in our region.

Genetic diversity of Staphylococcus aureus strains from a teaching hospital in Isfahan, Iran: The emergence of MRSA ST639- SCCmec III and ST343- SCCmec III.

BACKGROUND AND OBJECTIVES: Staphylococcus aureus is an important pathogen that can be colonized in the nose and increase the risk of spreading infections in hospitals. The present study aimed at determining the phenotypic and genotypic characterization of S. aureus strains isolated from patients and healthcare workers (HCWs) from a teaching hospital in Isfahan, Iran. MATERIALS AND METHODS: This cross-sectional study was performed on 262 nasal swabs and 23 clinical isolates that were collected from a teaching hospital during February and April 2016. Staphylococcal cassette chromosome mec (SCCmec) and multilocus sequence typing (MLST) were performed for selected isolates. RESULTS: Overall, 23% and 18% of healthcare workers and patients were carriers, respectively. Methicillin-resistant S. aureus (MRSA) rate was 13%, 33% and 52% in nasal HCWs, nasal patients, and clinical samples, respectively. The molecular typing of MRSA isolates revealed that the most common SCCmec type is SCCmec type III (88%). The highest rate of resistance was observed against tetracycline and erythromycin, with 48.7%. The most frequently detected toxin genes among S. aureus isolates were hla (99%) and sea (44%), moreover, pvl genes were detected in (40%) of MRSA isolates. The results of MLST showed 7 different sequence types (STs): ST859 (2/9), ST6 (2/9), ST639 (1/9), ST343 (1/9), ST239 (1/9), ST291 (1/9) and ST25 (1/9). CONCLUSION: Our results revealed that ST clones associated with healthcare-associated MRSA (HA-MRSA) are actively circulating among nasal carriage in our healthcare setting, and thus, effective infection control policies are needed to reduce nasal carriage in healthcare settings.

Biofilm Formation in Nonmultidrug-resistant Escherichia coli Isolated from Patients with Urinary Tract Infection in Isfahan, Iran.

BACKGROUND: Escherichia coli is a Gram-negative, opportunistic human pathogen in which increasing antibiotic resistance is a great concern for continued human survival. Although biofilm formation is a mechanism that helps E. coli to survive in unfavorable conditions, according to the importance of biofilm formation in developing the antibiotic resistance here, we studied the relation between antibiotic resistance and in vitro qualitative rating method biofilm formation in E. coli isolated from patients with urinary tract infection (UTI). MATERIALS AND METHODS: The clinical isolates of E. coli (n = 100) were collected from urine of patients with UTI attending Isfahan Alzahra hospital. The strains were confirmed as E. coli using biochemical tests and molecular method. The Kirby-Bauer disk diffusion tests were done according to the Clinical and Laboratory Standards Institute protocol, and the biofilm synthesis was performed by microplate method. The binary logistic test was applied and P < 0.05 was considered statistically significant. RESULTS: Our results showed a high outbreak of multidrug-resistant (MDR) E. coli strains (73%) and the highest resistance was observed toward ampicillin. The prevalence of biofilm producer isolates was 80% that 29% produced strong biofilm. The distribution of non-MDR isolates was high among strong biofilm producers, which shows a significant negative correlation between biofilm production and MDR pattern (P < 0.001). CONCLUSIONS: We found a negative correlation between MDR phenotype and biofilm formation capacity. This transmits the concept that more antibiotic susceptibility of strong biofilm producers may be due to the reduced exposure to multiple antibiotics.

Prevalence of CTX-M and TEM ß-lactamases in Klebsiella pneumoniae Isolates from Patients with Urinary Tract Infection, Al-Zahra Hospital, Isfahan, Iran.

BACKGROUND: Extended-spectrum ß-lactamase (ESBL)-producing is a significant resistant mechanism to ß-lactams in Enterobacteriaceae, especially in Klebsiella pneumoniae. The main objectives of this study were to genetically characterize urinary clinical isolates of K. pneumoniae through the investigating of blaTEM, blaCTX-M and using molecular typing by Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) method. We also determined the frequency of antibiotic resistance of K. pneumoniae strains to characterize the ß-lactamases included. MATERIALS AND METHODS: A cross-sectional study was carried out to evaluate 98 strains of K. pneumoniae isolated from urine culture of outpatients referred to Al-Zahra Hospital, Isfahan, Iran. Antibiotic susceptibility testing was performed using Kirby-Bauer's method. Screening of ESBLs was carried out using double-disk screening test. PCR technique was performed to detect TEM and CTX-M genes. The total DNA of each strain was tested by ERIC-PCR. RESULTS: In 98 K. pneumoniae studied clinical isolates, 25.5% were ESBL producing and 44.9% multidrug-resistant (MDR). From 25 ESBL isolates, 23 (92%) cases showed MDR phenotype. In ESBL producing isolates, 23 (92%) were blaCTX-M and 19 (76%) blaTEM positive. The antimicrobial drug susceptibilities of ESBL isolates indicated high resistant rates for cefotaxime and ceftazidime. All 25 ESBL producing isolates were resistant to cefotaxime. Complex patterns of fingerprints isolates showed that 36% of the isolates were belonged to the cluster no 5. CONCLUSION: This study revealed high antimicrobial resistance rates among ESBL isolates which can lead to various health difficulties. Epidemiological data collection from patients is recommended to develop the strategies to manage antibiotic resistance.

Molecular characteristics and antibiotic resistance pattern of Staphylococcus aureus nasal carriage in tertiary care hospitals of Isfahan, Iran.

Carriage of S. aureus in the anterior nares seems to play a significant role in the pathogenesis of infection. This study aimed to determine the molecular characteristics and antibiotic susceptibility pattern of S. aureus isolates obtained from the nasal carriage of health care workers (HCWs). This study was performed during July 2014 to July 2015 at three tertiary care hospitals. Nasal samples were collected from the nasal cavity of HCWs. Standard microbiological methods were used for identification of S. aureus isolates. Antibiotic susceptibility pattern was determined by the disc diffusion method. Determination of SCCmec typing and virulence genes was performed by the PCR method. From the isolates of 340 nasal swab samples of HCWs, 65 S. aureus strains (19%) including 22 (33.8%) MRSA were isolated. The highest sensitivity for MRSA isolates was towards vancomycin and rifampicin, each with 90.9%. Overall, 17% (11/65) and 92.3% (60/65) of S. aureus isolates were positive for pvl and hla genes, respectively. The rates of SCCmec types II, III, IV, V and I among MRSA isolates were 36.4 %, 22.7 %, 22.7 %, 9.1% and 4.5% respectively. The results of the present study indicate that S. aureus nasal carriage with potential virulence ability still remains a significant healthcare problem, especially in hospital environments.

Screening, isolation and molecular identification of biodegrading mycobacteria from Iranian ecosystems and analysis of their biodegradation activity.

Anthropogenic origin pollutants including pesticides, heavy metals, pharmaceuticals and industry chemicals impose many risks to human health and environment and bioremediation has been considered the strategy of choice to reduce the risk of hazardous chemicals. In the current study, we aimed to screen and characterize mycobacteria from the diverse range of Iranian aquatic and terrestrial ecosystems with harsh and unfavorable environmental conditions that can be utilized for biodegradation of target pollutants. Mycobacteria were isolated from a collection of 90 environmental samples and identified to the species level using conventional microbiological and molecular methods including the PCR amplification of hsp65 and sequence analysis of, 16S rRNA genetic markers. The growth rate of the isolates in presence of pollutants, chromatography, Gibbs and turbidometric methods were used to assess their biodegradation activity. A total of 39 mycobacterial isolates (43.3%) were recovered from 90 samples that belonged to 21 various species consisting of M. fortuitum; 6 isolates, M. flavescens and M. paragordonae; 4 isolates each, M. monacense, M. fredriksbergense and M. aurum; 2 isolates each, 7 single isolates of M. conceptionense, M. porcinum, M. simiae, M. celeriflavum, M. novocastrense, M. neoaurum, M. obuense and 12 isolates that belonged to 8 unknown potentially novel mycobacterial species. The isolates were categorized in three groups based on their bioremediation activity, i.e., 5 (12.8%) organisms without biodegradation activity, 20 (51.2%) organisms with previously reported biodegradation activity, and 14 (35.9%) organisms that showed biodegradation activity but not previously reported. Our results showed that the Iranian ecosystems harbor a good reservoir of diverse mycobacterial species with biodegrading potentiality for neutralizing environmental chemical pollutants.

Phenotypic and genotypic characterization of macrolide resistance among Staphylococcus aureus isolates in Isfahan, Iran.

BACKGROUND AND OBJECTIVES: Macrolide, lincosamide and streptogramin B (MLS B) are noteworthy antibiotics for the treatment of Staphylococcus aureus infections. The purpose of this study, was to determine the phenotypic and genotypic characterization of macrolide resistance, among S. aureus, isolated from clinical samples and nasal swabs. MATERIALS AND METHODS: Totally, 162 non-duplicate S. aureus isolates were collected from clinical samples and nasal swabs, from patients and healthcare workers (HCWs), between March 2016 and September 2016, at four teaching hospitals in Isfahan. The antibiotic resistance profile was determined using disk diffusion test and the presence of resistance genes was detected, using PCR. RESULTS: Of 162 S. aureus isolates, 43.8% (71/162) and 34% (55/162) isolates were erythromycin-resistant and methicillin-resistant S. aureus (MRSA), respectively. The prevalence of constitutive MLS B (cMLS B), inducible MLS B (iMLS B), macrolide-streptogramin B-resistant (MS B) and lincosamide-streptogramin-A resistance (LS A) phenotype was 32%, 6%, 6% and 2%, respectively. The most common erythromycin resistance genes, in S. aureus isolates were ermC (35.2%), followed by ermA (20.4%) and msrA (17.3%). Meanwhile, msrA was detected in 43.6% of MRSA isolates. The frequency of coexistence of ermA+ermC+msrA, in S. aureus isolates was 7% and it was only detected in MRSA isolates. CONCLUSION: In the current study, cMLS B phenotype was the most common erythromycin resistance pattern and ermC was the most prevalent gene in erythromycin-resistant isolates. The results revealed that the various mechanisms of erythromycin resistance are expanding in Isfahan.

Nasal carriage screening of community-associated methicillin resistant Staphylococcus aureus in healthy children of a developing country.

BACKGROUND: The rapid emergence and spread of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has raised considerable public health concern in both developed and developing countries. The current study aimed to address the extent of this phenomenon in healthy preschool children of a developing country. MATERIALS AND METHODS: We conducted a prospective study from April 2013 to March 2014 on 410 healthy 2-6 years old preschool children in Isfahan, Iran. Demographic medical data and nasal samples were collected from the participating children. Isolates were identified as S. aureus and MRSA based on microbiological and molecular tests, including the presence of eap and mecA genes. RESULTS: The overall prevalence of S. aureus and CA-MRSA nasal carriage was 28% (115/410) and 6.1% (25/410), respectively. The identity of isolates was confirmed by molecular assay. The factors that were independently associated with nasal carriage of S. aureus were: Children crowding in day-care nurseries and income level of families. A total of 20/90 (22.2%) of methicillin-susceptible S. aureus and all 25 CA-MRSA displayed multiple drug resistance to 3-8 antibiotics. CONCLUSIONS: The current report reflects issues and concerns that the high rate of colonization by CA-MRSA in Iranian healthy children provides obliging evidence that MRSA have established a foothold in the community and are emerging as important health threatening pathogens. It is suggested that we need more effective infection control measures to prevent transmission of nasal CA-MRSA in healthy preschool children.

Nasal colonization in children with community acquired methicillin-resistant Staphylococcus aureus.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is a frequent cause of infections. The changing epidemiology of MRSA became evident in the 1990s when CA-MRSA cases were first reported. Nasal carriage of CA-MRSA is associated with an increased risk for development of infections in various populations. MATERIALS AND METHODS: Anterior nares culture for the presence of methicillin-susceptible Staphylococcus aureus (MSSA) and MRSA was taken from 345 children attending kindergartens, who didn't have any known risk factor for MRSA colonization. Also, children demographic variables were recorded. Identification of SA and community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) with standard microbiological test was performed. Finally, the susceptibility of isolated to various antibiotics determined. The data were analyzed with Whonet 5.6 software. RESULTS: Of 345 children, 20 children (5.8%) were colonized with CA-MRSA, 86 children (24.9%) with MSSA and 239 cases (69.3%) didn't have SA colonization. The highest rate of MSSA and MRSA colonization was obtained at the age of 6 years. The frequency distribution of SA (MSSA and MRSA) colonization prevalence didn't have any significant differences based on age, gender and the admission time (P > 0.05); but it was significantly different in the urban areas (P < 0.001). The lowest resistance rate of CA-MRSA isolates, with a frequency of 10%, was detected with gentamicin, rifampin, and trimethoprim-sulfamethoxazole. CONCLUSIONS: In summary, CA-MRSA colonization was observed in child care centers remarkably. Therefore, by facing various infections due to SA especially in areas of low socio-economic status, it must be considered. Based on antibiogram test, empirical treatment with rifampin, gentamicin and ciprofloxacin is recommended during CA-MRSA infections.

Application of the Random Amplified Polymorphic DNA (RAPD) Fingerprinting to Analyze Genetic Variation in Community Associated-Methicillin Resistant Staphylococcus Aureus (CA-MRSA) Isolates in Iran.

The aim of this study was to apply RAPD technique to analyze the genetic variability among the Iranian CA-MRSA isolates.The RAPD amplification was implemented on 25 strains isolated from the anterior nares of 410 healthy children using four randomly selected oligonucleotide primers from the stocks available in our laboratory, including the primers 1254, GE6, OLP6 and OLP13 from our stock. The amplified PCR products were detected on a 1.5% agarose gel and subjected to further analysis to establish the band profiles and genetic relationships using the Gel Compar® program.The Iranian CA-MRSA isolates produced distinct RAPD patterns which varied based on the primer used, however, the primer 1254 revealed highly polymorphic patterns consisting 5 discernable RAPD types (RT), "RT1" (12, 48%), "RT2" (8, 32%), "RT3" (3, 12%), and "RT4 and RT5", (a single RAPD type each, 4%). Phylogenetic analysis based on RAPD profiles divided most of the CA-MRSA isolates into 2 distinct but related RAPD clusters, a small group and two single unrelated RAPD types.This study shows that the simple and cost-effective but rather difficult to optimize RAPD fingerprinting could be used to evaluate genetic and epidemiological relationships of CA-MRSA isolates on condition that the patterns are obtained from carefully optimized laboratory tests.

Study on Prevalence, Antibiotic Susceptibility, and tuf Gene Sequence-Based Genotyping of Species-Level of Coagulase-Negative Staphylococcus Isolated From Keratitis Caused by Using Soft Contact Lenses.

OBJECTIVE: To study on antibiotic susceptibility and identify coagulase-negative Staphylococcus (CoNS) species based on tuf gene sequencing from keratitis followed by using soft contact lenses in Isfahan, Iran, 2013. METHODS: This study examined 77 keratitis cases. The samples were cultured and the isolation of CoNS was done by phenotypic tests, and in vitro sensitivity testing was done by Kirby-Bauer disk diffusion susceptibility method. RESULTS: Thirty-eight of isolates were conveniently identified as CoNS. In this study, 27 (71.1%), 21 (55.3%), and 16 (42.1%) were resistant to penicillin, erythromycin, and tetracycline, respectively. One hundred percent of isolates were sensitive to gentamicin, and 36 (94.7%) and 33 (86.8%) of isolates were sensitive to chloramphenicol and ciprofloxacin, respectively. Also, resistances to cefoxitin were 7 (18.4%). Analysis of tuf gene proved to be discriminative and sensitive in which all the isolates were identified with 99.0% similarity to reference strains, and Staphylococcus epidermidis had the highest prevalence among other species. CONCLUSIONS: Results of this study showed that CoNS are the most common agents causing contact lens-associated microbial keratitis, and the tuf gene sequencing analysis is a reliable method for distinguishing CoNS species. Also gentamycin, chloramphenicol, and ciprofloxacin are more effective than the other antibacterial agents against these types of bacteria.

Antibacterial effect of triantibiotic mixture, chlorhexidine gel, and two natural materials Propolis and Aloe vera against Enterococcus faecalis: An ex vivo study.

BACKGROUND: The aim of this ex vivo study was to compare the antimicrobial effect of triantibiotic paste, 0.2% chlorhexidine gel, Propolis and Aloe vera on Enterococcus faecalis in deep dentin. MATERIALS AND METHODS: Ninety fresh extracted single-rooted teeth were used in a dentin block model. Seventy-five teeth were infected with E. faecalis and divided into four experimental groups (n = 15). Experimental groups were treated with triantibiotic mixture with distilled water, 0.2% chlorhexidine gel, 70% ethanol + Propolis and Aloe vera. Fifteen teeth treated with distilled water as the positive control and 15 samples, free of bacterial contamination, were considered as the negative control. Gates-Glidden drill #4 was used for removal of surface dentin and Gates-Glidden drill #5 was used to collect samples of deep dentin. The samples were prepared and colony-forming units were counted. Data were analyzed by one-way ANOVA and post hoc Tukey tests. Statistical significance was defined at P < 0.05. RESULTS: Triantibiotic mixture group exhibited the least bacterial growth. However, the rate of bacterial growth showed no significant differences between chlorhexidine and Propolis groups (P > 0.05). Aloe vera had antibacterial effects on E. faecalis, but in comparison with other medicaments, it was less effective (P < 0.05). CONCLUSION: This experimental study showed that triantibiotic mixture, 0.2% chlorhexidine gel, Propolis and Aleo vera were relatively effective against E. faecalis. All the intracanal medicements had similar effects on E. faecalis in deep dentin except for Aloe vera.

Frequency assessment of ß-lactamase enzymes in Escherichia coli and Klebsiella isolates in patients with urinary tract infection.

BACKGROUND: Production of ß-lactamase enzymes is the most common and important mechanism of resistance in Gram-negative bacteria. The objective of this study was to assess frequency of three main ß-lactamase enzymes, including extended spectrum ß-lactamases (ESBLs), metallo-ß-lactamase (MBL), and Klebsiella pneumoniae carbapenemase (KPC) enzymes in Escherichia coli and Klebsiella spp. isolated from nosocomial and community urinary tract infections (UTI). MATERIALS AND METHODS: In a cross-sectional study from March to December 2012, midstream urine samples were obtained from patients suspicious of UTI who were hospitalized or referred to Al-Zahra Hospital, Isfahan, Iran. Samples were cultured and E. coli and Klebsiella spp. were isolated. Prevalence of ESBLs, KPC, and MBLs producing E. coli and Klebsiella spp. were studied by double-disk (combined-disk), the modified Hodge test and imipenem-ethylenediaminetetraacetic acid combined disc methods respectively. In addition, their antimicrobial susceptibility patterns determined and resistant to carbapenem drugs confirmed by minimum inhibitory concentrations based on E-test method. RESULTS: A total of 1080 E. coli and 484 Klebsiella strains were isolated during study period. Among 720 E. coli and 384 Klebsiella isolates from hospitalized patients, 300 (41.7%) and 198 (51.5%) were ESBLs producers, respectively. In out-patients samples, the rate of ESBLs production was 25% (90/360) and 40% (40/100) in E. coli and Klebsiella isolates, respectively. Prevalence of MBLs producing in hospital E. coli and Klebsiella isolates were 0.3% (2/720) and 2.6% (10/384), and for KPC data were 1.4% (10/720) and 48.4% (186/384), respectively. No MBLs and KPC producing isolate was seen in non-hospital E. coli and Klebsiella isolates except for one non-hospital KPC producing Klebsiella isolate. CONCLUSION: The result of our study showed high prevalence of ESBLs and KPC, but low prevalence of MBLs in cultured bacteria from urine samples of patients with acute UTI. In addition, KPC was the main carbapenem resistance mechanism in Klebsiella and E. coli isolates.

Antimicrobial activity of a UV-stable bacteriocin-like inhibitory substance (BLIS) produced by Enterococcus faecium strain DSH20 against vancomycin-resistant Enterococcus (VRE) strains.

BACKGROUND/PURPOSE: The narrow spectrum of action of most bacteriocins is an important limitation for their application as antimicrobial agents. The current study describes a novel bacteriocin-like inhibitory substance (BLIS) that display extended spectrum antimicrobial activity against vancomycin-resistant Enterococcus (VRE) strains. METHODS: Acquired resistance profiles of Enterococcus isolates determined based on the European Centre for Disease Prevention and Control (ECDC) and the Centers for Disease Control and Prevention (CDC) definition as multidrug-resistant (MDR), extensively drug-resistant (XDR) and pandrug resistant (PDR). BLIS activity of Enterococcus isolates was investigated against Enterococcus faecalis (E. faecalis) ATCC 29212 as the indicator strain and clinical isolates including VRE, methicillin resistant Staphylococcus aureus (MRSA) and Gram-negative bacteria containing Pseudomonas aeruginosa (P. aeruginosa), Klebsiella, Acinetobacter, and Escherichia coli (E. coli). RESULTS: Among 273 Enterococcus isolates, 27 and 2 VRE isolates, respectively, were XDR and PDR and eight isolates had BLIS activity against the indicator strain. One of these isolates, identified as E. faecium strain DSH20 based on its phenotypical and biochemical properties, as well as its 16S rRNA gene sequence, had potent BLIS production against all 29 VRE strains, but had no activity against MRSA, P. aeruginosa, Klebsiella, Acinetobacter, and E. coli strains. It was heat stable up to 121°C for 15 minutes (autoclave condition), active within the pH range of 3-9 and had UV stability, but its activity disappeared by treatment with proteinase K, pepsin, and trypsin, demonstrating its proteinaceous nature. It was designated as an approximately 35kDa peptide using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method. CONCLUSION: This peptide is a potential agent for use as an alternative antibacterial agent for the treatment of drug-resistant strains of VRE infection.

Phenotypic and Molecular Characterization of Extended-Spectrum ß-Lactamase Produced by Escherichia coli, and Klebsiella pneumoniae Isolates in an Educational Hospital.

BACKGROUND: Extended-spectrum beta-lactamases (ESBLs) are a group of enzymes that hydrolyze antibiotics, including those containing new cephalosporins, and they are found in a significant percentage of Escherichia coli and Klebsiella pneumoniae strains. With the widespread use of antibiotics, difficulties with infection therapy caused by drug resistant organisms, especially those that have acquired resistance to beta-lactams, such as broad-spectrum cephalosporins, have amplified the above-mentioned organisms. OBJECTIVES: This study was conducted to characterize ESBLs among E. coli and K. pneumonia isolates by molecular and phenotypic methods. MATERIALS AND METHODS: Different strains of E. coli and K. pneumonia were collected from patients with urinary tract infections. The ESBL phenotype was determined by a double disk diffusion test (DDDT). In addition, polymerase chain reaction (PCR) analysis specific for ß-lactamase genes of the TEM and SHV family was carried out. The PCR products were run on agarose and examined for DNA bands. RESULTS: A total of 245 E. coli and 55 K. pneumonia strains were isolated from different samples. In total, 128 of the 300 isolates were confirmed as potential ESBLs producers as follows: 107 (43.67%) E. coli and 21 (38.18%) K. pneumonia. ESBLs genes were found in 24 isolates (18.75%): 21 E. coli and 3 K. pneumonia isolates. The TEM gene was present in 13 (12.14%) E. coli strains, but it was not detected in K. pneumonia. In addition, the SHV gene was present in 8 (7.47%) E. coli and 3 (14.28%) K. pneumonia isolates. Five (4.67%) of the E. coli isolates harbored both TEM and SHV genes. All isolates (100%) were susceptible to imipenem. The lowest rates of resistance to other antibiotics were observed for; piperacillin-tazobactam (6.25%), amikacin (12.5%) and gentamicin (14.84%). The rates of resistance to other antibiotics were as follow: nitrofurantoin (16.4%), nalidixic acid (23.43), co-trimoxazole (25%), cefepime (32%), ciprofloxacin (55.46%), ampicillin (69.53%), ceftazidime (100%), and cefotaxime (100%). CONCLUSIONS: The results of this study indicate the widespread prevalence of ESBLs and multiple antibiotic resistance in E. coli and K. pneumoniae. Therefore, beta-lactam antibiotics and beta-lactamase inhibitors or carbapenems should be prescribed based on an antibacterial susceptibility test.

Susceptibility pattern of E. coli-associated urinary tract infection (UTI): a comparison of spinal cord injury-related and nosocomial UTI.

BACKGROUND: Evolving resistant uropathogens are associated with elevated risk of morbidity and mortality both in nosocomial and in spinal cord injury (SCI)-associated urinary tract infections (UTIs). Clinico-epidemiological studies are therefore essential to facilitate empirical therapy and decrease treatment time. MATERIAL/METHODS: This study was performed at Al-Zahra Hospital, Isfahan University of Medical Sciences, Isfahan, Iran. All E. coli-infected UTI hospitalized patients, except those with symptoms of UTI prior to admission or in the first 48 hours of hospitalization, were included in this study as group A. All E. coli-infected SCI-associated UTIs were included in group B. Urine samples were cultured and the MICs of nine antibiotics on the isolated bacteria were determined by the gradient concentration method with E. coli ATCC25922 as the standard control. RESULTS: Antibiotic-resistant uropathogens were more prevalent among the nosocomial than the SCI-associated UTIs. The antibiotic-resistance rates of isolated E. coli in the nosocomial and SCI-associated UTIs were 13.9% and 5.9% for amikacin, 65% and 40.5% for ceftazidime, 69.8% and 32.4% for ceftriaxone, 50% and 41.2% for ciprofloxacin, 56.9% and 13.6% for gentamicine, 2.1% and 0% for imipenem, 3.9% and 0% for meropenem, and 73.6% and 63.6% for trimethoprim/sulfamethoxazole, respectively. Only resistance to nalidixic acid was mildly higher in SCI E. coli. The MIC50 of all the antibiotics except nalidixic acid was higher in nosocomial E. coli. CONCLUSIONS: Despite more risk factors for antibiotic-resistant E. coli in SCI patients, antibiotic-resistant E. coli was observed more in nosocomial UTIs, which may be attributable to the hospital environment.