Molecular typing of clinical multidrug-resistant Klebsiella pneumoniae isolates.

INTRODUCTION: Klebsiella pneumoniae is an opportunistic pathogen which is an important cause of hospital-acquired and antibiotic resistance infections. Therefore, this study aimed to determine the frequency of resistance to antibiotics, as well as the molecular typing of the associated isolates, and compare multiple-locus VNTR analysis (MLVA) and Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR) methods to specify the degree to which distinctions can be separated from each other. METHODS AND MATERIALS: One hundred K. pneumoniae isolates were obtained from different sources of infections from patients admitted to hospitals. Antibiotic susceptibility testing was then performed by applying the Kirby-Bauer disk diffusion method. Typing of K. pneumoniae was done by utilizing MLVA and ERIC-PCR methods. RESULTS: Eighty-six multidrug-resistant (MDR) K. pneumoniae isolates were identified, which resistance to ampicillin, trimethoprim/sulfamethoxazole, and ceftriaxone was the most frequent in the considered isolates (100, 93, and 93%, respectively). A total of 50 different antibiotic susceptibility patterns were observed among the MDR K. pneumonia, with the most frequent pattern being resistance to all antibiotics (12.79%) and resistance to all antibiotics except amikacin (10.47%). The isolates were then divided into 37 different MLVA types and seven clonal complexes were obtained from the minimum spanning tree analysis. Finally, the isolates were assigned to 38 different ERIC types. The discriminatory power of MLVA and ERIC methods also showed a value of 0.958, and 0.974. CONCLUSION: Both PCR-typing methods with phenotypic patterns can be useful for the epidemiological typing of K. pneumoniae isolates with the highest performance in discriminating isolates.

Molecular Typing of Multidrug-Resistant Acinetobacter baumannii Isolates from Clinical Specimens by ERIC-PCR and MLVA.

Acinetobacter baumannii, a Gram-negative and oxidase-negative bacterium, is a major cause of nosocomial infections, leading to high mortality rates in hospitalized patients. The use of 2 prominent molecular typing methods (i.e., enterobacterial repetitive intergenic consensus-polymerase chain reaction [ERIC-PCR] and multiple-locus variable-number tandem repeat [VNTR] analysis [MLVA]) for genotyping A. baumannii isolates has proven to be an effective approach in assessing the clonal relation of these isolates and managing their outbreaks. A total of 100 A. baumannii isolates were collected from immunocompromised patients hospitalized in the intensive care unit (ICU) of a hospital in Zanjan City, Iran. Their antibiotic resistance ability (especially aminoglycoside resistance) was studied by disc diffusion tests. The genetic typing of A. baumannii was studied using ERIC-PCR and MLVA methods. All isolates were resistant to 3 or more antibiotics and regarded as multidrug-resistant (MDR). Additionally, 32% of the isolates were resistant to all antibiotics tested, and 91% were extensively drug-resistant (XDR). The increased rate of aminoglycoside-resistant A. baumannii in ICU patients, with an increased incidence of aminoglycoside-modifying enzymes of aac (6')-Ib, ant (3″)-I, and aph (2″)-Id. ERIC-PCR has likewise shown an increased level of diversity in A. baumannii isolates. According to the ERIC-PCR patterns, isolates were classified as 4 clusters, while according to the MLVA patterns, isolates were classified as 9 distinct clusters. ERIC-PCR and MLVA assays serve as useful genotyping methods to assess the genetic variety or clonal relatedness of A. baumannii isolates.

Antimicrobial Activity of Silver Nanoparticles Synthesized by the Green Method Using Rhus coriaria L. Extract Against Oral Pathogenic Microorganisms.

Background: Oral health is part of general health. Dental caries is the most common chronic disease worldwide. Considering the significance of plaque control, complications of chemical agents, and the optimal antimicrobial efficacy of nanoparticles, this study aimed to assess the antimicrobial activity of silver nanoparticles (AgNPs) synthesized by the green method using Rhus coriaria L. extract against oral pathogenic microorganisms. Methods: In this in vitro experimental study, Rhus coriaria fruit was dried at room temperature. It was then ground, and its aqueous extract was obtained by the maceration technique. The effects of AgNPs synthesized by the green method in different concentrations were evaluated against Enterococcus faecalis (E. faecalis), Streptococcus mutans (S. mutans), Streptococcus sobrinus (S. sobrinus), Lactobacillus acidophilus (L. acidophilus), and Streptococcus salivarius (S. salivarius) using the well-plate technique. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were also calculated. Data were analyzed by the Wilcoxon test, Kruskal-Wallis test, and Mann-Whitney test. Results: The MIC values were 1024 µg/mL for S. mutans and E. faecalis, and 512 µg/mL for S. sobrinus, S. salivarius, and L. acidophilus. The resistance of S. mutans and E. faecalis was higher than that of S. sobrinus, S. salivarius and L. acidophilus. According to the growth inhibition zones and MBC test results, S. salivarius had the highest resistance to AgNPs followed by L. acidophilus, S. sobrinus, S. mutans, and E. faecalis. Conclusion: AgNPs synthesized by the green method using Rhus coriaria extract was effective against oral pathogenic microorganisms. Thus, they may be used in the formulation of mouthwash and toothpaste.

High frequency of enterotoxigenic Bacteroides fragilis and Enterococcus faecalis in the paraffin-embedded tissues of Iranian colorectal cancer patients.

BACKGROUND: The association between specific bacteria and colorectal cancer (CRC) has been proposed. Only a few studies have, however, investigated this relationship directly in colorectal tissue with conflicting results. So, we aimed to quantitate Streptococcus gallolyticus, Fusobacterium spp, Enterococcus faecalis and enterotoxigenic Bacteroides fragilis (ETBF) in formalin-fixed and paraffin-embedded (FFPE) colorectal tissue samples of Iranian CRC patients and healthy controls. METHODS: A total of 80 FFPE colorectal tissue samples of CRC patients (n = 40) and healthy controls (n = 40) were investigated for the presence and copy number of above bacterial species using quantitative PCR. Relative quantification was determined using ΔΔCT method and expressed as relative fold difference compared to reference gene. RESULTS: Relative abundance and copy number of E. faecalis and ETBF were significantly higher in CRC samples compared to control group. E. faecalis was more prevalent than ETBF in tumor samples. Frequency of ETBF and E. faecalis in late stages (III/IV) of cancer was significantly higher than early stages (I/II). We did not detect a significant difference in abundance of S. gallolyticus and Fusobacterium spp between two groups. CONCLUSION: Our study revealed the higher concentration of E. faecalis and ETBF in FFPE samples of CRC patients than controls. However, additional investigations on fecal and fresh colorectal cancer tissue samples are required to substantiate this correlation.

Genetic diversity of colistin resistance Nosocomial Acinetobacter baumannii strains from Iran.

BACKGROUND: Drug-resistant Acinetobacter baumannii is a global health problem since its ability to acquire new resistance mechanisms. Here, we aimed to determine the association of common types of A. baumannii and assess their drug resistance of A. baumannii and contribution of integrons (Ints) and oxacillinase genes in Zanjan, Iran. MATERIALS AND METHODS: Among 68 isolated Acinetobacters from patients, 48 isolates were A. baumannii strains. Antibiotic susceptibility pattern and colistin resistance were determined by disk diffusion and broth microdilution, respectively. The presence of Int I, II, III, and oxacillinase genes examined using polymerase chain reaction. The clonal relationship of clinical isolates of A. baumannii determined by Pulsed Field Gel Electrophoresis method. RESULTS: The results showed the highest antibiotic susceptibility (58%) for colistin. 96% of isolates were considered as multidrug resistant, and 46% as extensively drug resistant, and 16% as pandrug resistant. Frequencies of Int I, II, III resistance genes were 60%, 28%, and 0%, respectively, and 12% of strains had no isoform of Ints. Frequencies of Carbapenem resistance genes were 74%, 24%, 100%, and 4% for blaOXA-23, blaOXA-24, blaOXA-51, and blaOXA-58, respectively. The above samples were group into 26 pulsotypes. CONCLUSION: The studied A. baumannii strains had several resistance genes, and the colistin resistance showed an extraordinary ascending tendency that could be a severe issue in nosocomial infections, and the presence of high genetic diversity indicated a variation in A. baumannii strains and possibly a variety of sources of contamination or infection.

The association between fecal enterotoxigenic B. fragilis with colorectal cancer.

BACKGROUND: Enterotoxigenic Bacteroides fragilis (ETBF) is an enterotoxin-producing bacterium that possibily has a role in the occurrence and progression of colorectal cancer (CRC) by modulating the mucosal immune response and inducing epithelial cell changes. The aim of this study was to investigate the frequency of ETBF in stool samples of CRC patients and healthy volunteers. METHODS: A total of 60 stool samples from confirmed CRC patients and 60 stool samples from healthy volunteers with no personal or familial history or diagnosis of colorectal disease were collected. Stool samples were screened for direct detection of B. fragilis using PCR targeting the marker genes of neu and bft. Enterotoxin isotypes bft-1, bft-2 and bft-3 were also detected in B. fragilis positive samples. RESULTS: The frequency of B. fragilis among CRC and control cases was 58.3 and 26.6%, respectively (P < 0.05). The rate of bft gene in CRC cases was significantly higher than in controls (P < 0.05). Also, the presence of bft gene in CRC patients stage III was significantly higher than stages I and II (P < 0.05). Enterotoxin isotype bft-2 was detected with higher frequency among CRC patients than healthy control (P < 0.05). CONCLUSION: Our results show the association between fecal ETBF and CRC, and we suggest that detection of ETBF may be a potential marker for colorectal cancer diagnosis. However, additional investigations on tumor and paired normal tissue samples are required to substantiate this possible correlation.

High incidence of virulence determinants, aminoglycoside and vancomycin resistance in enterococci isolated from hospitalized patients in Northwest Iran.

BACKGROUND: Multidrug resistant (MDR) enterococci are important nosocomial pathogens causing serious problem in hospitalized patients. The aim of present study was to investigate the frequency of high-level aminoglycoside-resistant and vancomycin-resistant enterococci (VRE) and virulence encoding genes in enterococci isolated from hospitalized patients. METHODS: A total of 100 enterococci isolated from urine samples of hospitalized patients with symptomatic urinary tract infections were investigated for antimicrobial susceptibility, the frequency of aminoglycoside and vancomycin resistance genes (including aac (6')-Ie-aph (2")-Ia, aph (3')-IIIa, ant (4')-Ia, aph (2")-Ic, aph (2")-Ib, aph (2")-Id, ant (3″)-III, ant (6')-Ia, vanA, vanB and vanC) and virulence encoding genes (including gelE, PAI, esp, ace, cyl, hyl and sprE). RESULTS: Enterococcus faecalis species was identified as predominant enterococci (69%), followed by "other" Enterococcus species (21%) and E. faecium (10%). Ninety three percent of isolates were resistant to one or more antimicrobial agents, with the most frequent resistance found against tetracycline (86%), ciprofloxacin (73%) and quinupristin-dalfopristin (53%). Gentamicin and streptomycin resistance were detected in 50 and 34% of isolates, respectively. The most prevalent aminoglycoside resistance genes were ant (3″)-III (78%) and aph (3')-IIIa (67%). Vancomycin resistance was detected in 21% of isolates. All E. faecium isolates carried vanA gene, whereas, the vanB gene was not detected in Enterococcus species. The most frequent virulence gene was ace (88.6%), followed by esp (67.1%), PAI (45.5%) and sprE (41.7%). CONCLUSION: Our study revealed the high frequency of gentamycin resistance and VRE in E. faecium isolates, with a high prevalence and heterogeneity of virulence and resistance genes. Due to high frequency of MDR enterococci, it seems that the appropriate surveillance and control measures are essential to prevent the emergence and transmission of these isolates in hospitals.

Virulence characteristics of multidrug resistant biofilm forming Acinetobacter baumannii isolated from intensive care unit patients.

BACKGROUND: Nosocomial infections and persistence of multidrug resistant biofilm forming Acinetobacter baumannii in hospitals has made it as a serious problem in healthcare settings worldwide. METHODS: A total of 100 A. baumannii clinical isolates from immunocompromised patients hospitalized in ICU were investigated for biofilm formation, the presence of biofilm related genes (bap, ompA, csuE, fimH, epsA, blaPER-1, bfmS, ptk, pgaB, csgA, kpsMII), integron characterization and molecular typing based on REP-PCR. RESULTS: All isolates were resistant to three or more categories of antibiotics and considered as multidrug resistant (MDR). A total of 32 isolates were resistant to all tested antibiotics and 91% were extensively drug-resistance (XDR). All isolates were able to produce biofilm and 58% of isolates showed strong ability to biofilm formation. All strong biofilm forming A. baumannii isolates were XDR. All A. baumannii isolates carried at least one biofilm related gene. The most prevalent gene was csuE (100%), followed by pgaB (98%), epsA and ptk (95%), bfmS (92%) and ompA (81%). 98% of isolates carried more than 4 biofilm related genes, simultaneously. Class I integron (67%) was more frequent in comparison with class II (10%) (P < 0.05). The REP-PCR patterns were classified as 8 types (A-H) and 21 subtypes. The A1 (23%) and C1 (15%) clusters were the most prevalent among A. baumannii isolates (P < 0.05). According to the REP-PCR patterns, 23% of all isolates had a clonal relatedness. CONCLUSION: Our study revealed the high frequency of biofilm forming XDR A. baumannii in ICU patients, with a high prevalence of biofilm related genes of csuE and pgaB. It seems that the appropriate surveillance and control measures are essential to prevent the emergence and transmission of XDR A. baumannii in our country.

Diversity of virulence genes in multidrug resistant Pseudomonas aeruginosa isolated from burn wound infections.

Multidrug resistant Pseudomonas aeruginosa has frequently been reported as the cause of nosocomial outbreaks of burn wound infections. The pathogenesis of P. aeruginosa is partly due to the production of several cell-associated and extracellular virulence factors. A total of 93 P. aeruginosa isolated from burn wound infections were investigated for antimicrobial susceptibility and distribution of virulence genes. All (100%) isolates were resistant to one or more antimicrobial agents. The most frequent resistance found against ampicillin (91.4%), co-trimoxazole (77.4%), gentamicin (68.8%), cefotaxime (50.5%), aztreonam and piperacillin (41.9%). A total of 88 (94.6%) isolates were resistant to at least three different classes of antimicrobial agents and considered as multidrug resistance MDR. All isolates carried at least two or more different virulence genes. The most prevalent virulence gene was toxA (97.8%), followed by plcH (96.7%), phzI (96.7%), exoY (93.1%) and phzII (90.3%). exoU was not detected in P. aeruginosa isolates. The frequency of pilB (17.2%), exoT (20.4%), pilA (24.7%) and phzS/phzH (27.9%) was lower than other virulence genes. Twenty nine (31.2%) isolates had simultaneously 8 virulence genes, 22 (23.7%) isolates had 6 virulence genes and 19 (20.4%) isolates had 7 virulence genes. All MDR isolates carried at least 5 virulence factors. These results indicate a high frequency and heterogeneity of virulence gene profiles among multidrug resistant P. aeruginosa isolates recovered from burn wound infections. Therefore, appropriate surveillance and control measures are essential to prevent the further spread of these isolates in hospitals.

Synergistic activity of sub-inhibitory concentrations of curcumin with ceftazidime and ciprofloxacin against Pseudomonas aeruginosa quorum sensing related genes and virulence traits.

Quorum sensing (QS) system in Pseudomonas aeruginosa may be an important target for pharmacological intervention. The present study aimed to investigate the synergetic activity of sub-MIC concentrations of curcumin (C) with ceftazidime (CAZ) and ciprofloxacin (CIP) against P. aeroginusa QS system. We determined the MIC and synergistic activity of C, CAZ and CIP against P. aeroginusa PAO1 using broth microdilution and checkerboard titration methods. The activity of sub-MIC (1/4 and 1/16 MIC) concentrations of C on the QS signal molecules was assessed using a reporter strain assay. The influence of sub-MIC of C, CAZ and CIP alone and in combination on motility and biofilm formation was also determined and confirmed by RT-PCR to test the expression of QS regulatory genes lasI, lasR, rhlI and rhlR. The addition of C decreased the MIC of CAZ and CIP. Curcumin showed synergistic effects with CAZ and additive activity with CIP. Treated PAO1 cultures in the presence of C showed significant reduction of signals C12-HSL and C4-HSL (P < 0.05). Sub-MIC concentrations (1/4 and 1/16 MIC) of C, CAZ and CIP alone and in combination significantly reduced swarming and twitching motilities and biofilm formation. Expression of QS regulatory genes lasI, lasR, rhlI, and rhlR using 1/4 MIC of C, CAZ and CIP alone and in combination was repressed significantly relative to untreated PAO1. Our results indicate that a combination of the sub-MIC concentration of C and CAZ exhibited synergism against P. aeroginusa QS system. This combination could lead to the development of a new combined therapy against P. aeruginosa.

Engineering a platelet-rich plasma-based multifunctional injectable hydrogel with photothermal, antibacterial, and antioxidant properties for skin regeneration.

Wound healing remains a significant challenge worldwide, necessitating the development of new wound dressings to aid in the healing process. This study presents a novel photothermally active hydrogel that contains platelet-rich plasma (PRP) for infected wound healing. The hydrogel was formed in a one pot synthesis approach by mixing alginate (Alg), gelatin (GT), polydopamine (PDA), and PRP, followed by the addition of CaCl2 as a cross-linker to prepare a multifunctional hydrogel (AGC-PRP-PDA). The hydrogel exhibited improved strength and good swelling properties. PDA nanoparticles (NPs) within the hydrogel endowed them with high photothermal properties and excellent antibacterial and antioxidant activities. Moreover, the hydrogels sustained the release of growth factors due to their ability to protect PRP. The hydrogels also exhibited good hemocompatibility and cytocompatibility, as well as high hemostatic properties. In animal experiments, the injectable hydrogels effectively filled irregular wounds and promoted infected wound healing by accelerating re-epithelialization, facilitating collagen deposition, and enhancing angiogenesis. The study also indicated that near-infrared light improved the healing process. Overall, these hydrogels with antibacterial, antioxidant, and hemostatic properties, as well as sustained growth factor release, show significant potential for skin regeneration in full-thickness, bacteria-infected wounds.

Phylogenetic groups and antimicrobial resistance characteristics of Escherichia coli strains isolated from clinical samples in North Iran.

BACKGROUND AND STUDY AIM: Extraintestinal pathogenic Escherichia coli (ExPEC) is one of the most common bacterial pathogens, which causes a remarkable amount of morbidity and mortality. This study was designed to determine the antibiotic resistance profiles, phylogenetic groups, and subgroup analyses among the ExPEC strains isolated from hospitalized patients in north Iran. PATIENTS AND METHODS: This cross-sectional investigation was conducted at five educational hospitals in Rasht in north Iran. Using standard microbiological tests, 150 E. coli isolates were identified. The antibiotic susceptibility pattern of all isolates was determined using the disk diffusion method. The double disk phenotypic confirmatory test was performed to detect extended-spectrum ß-lactamase (ESBL)-producing isolates. A triplex polymerase chain reaction (PCR) was performed to determine the phylogenetic group of each strain. RESULTS: The results of antibiogram pattern showed that E. coli isolates were mostly non-susceptible to ampicillin (79.3%), followed by nalidixic acid (75.3%) and cephalothin (70%), whereas nitrofurantoin (94.7%) was the most effective agent, followed by imipenem (92.7%). The rate of ESBL-producing isolates was 53.3% (80/150). Multiplex PCR screening revealed that the most common phylogroup was the B2 group (97 isolates; 64.6%), followed by the D group (34, 22.7%). In contrast, phylogroup analyses showed that B23 (50.7%) and D2 (16.4%) were the most common subgroups. CONCLUSIONS: Our findings indicated a considerable rate of antibiotic resistance and ESBL-producing isolates among E. coli strains isolated from clinical samples. Moreover, we reported a tendency that most isolates belonged to the B2 and D phylogroups. As a result, the detection of genotypic identical or similar isolates indicated that these isolates have an endurance capability in the hospital environment and could be transmitted among patients.

High frequency of enterotoxin encoding genes of Staphylococcus aureus isolated from food and clinical samples.

BACKGROUND: Staphylococcus aureus is recognized as an important cause of food poisoning related to the consumption of raw, undercooked, or mishandled foods worldwide. METHODS: A total of 90 individual meat samples and 200 clinical specimens were collected and investigated the frequency of S. aureus and classical enterotoxin genes. The samples were cultured on Baird-Parker and Mannitol salt agar and subjected for confirmatory biochemical tests and molecular detection of femA, sea, seb, sec, sed, and see genes. RESULTS: A total of 31 (34.5%) meat samples and 81 (40.5%) clinical specimens were positive for the presence of S. aureus. These isolates were detected with slightly higher frequency in clinical specimens than food samples (P> 0.05). Furthermore, the frequency of S. aureus in raw meat (23.4%) was higher than that in cooked meat samples (11.1%) (P< 0.05). Staphylococcal enterotoxin (SE) genes were identified in 18 (58.1%) of 31 meat isolates and 42 (51.8%) of 81 clinical isolates. The frequency of SE genes (except see) in meat isolates was slightly higher than that in clinical isolates (P> 0.05). We found sea and see genes with higher frequency than others in both meat and clinical samples. Furthermore, 55.5% of meat isolates and 38.1% of clinical isolates possessed more than one se gene. CONCLUSION: Detection of enterotoxigenic S. aureus in clinical and raw meat samples shows a probable risk for public health. Therefore, intensive and continuous monitoring of potentially pathogenic S. aureus is strongly recommended in order to evaluate the human health risk arising from food consumption.

Staphylococcus aureus Putative Vaccines Based on the Virulence Factors: A Mini-Review.

Since the 1960s, the frequency of methicillin-resistant Staphylococcus aureus as a recurrent cause of nosocomial infections has increased. Since multidrug-resistant Staphylococcus has overcome antimicrobial treatment, the development of putative vaccines based on virulence factors could be a great help in controlling the infections caused by bacteria and are actively being pursued in healthcare settings. This mini-review provides an overview of the recent progress in vaccine development, immunogenicity, and therapeutic features of some S. aureus macromolecules as putative vaccine candidates and their implications against human S. aureus-related infections. Based on the reviewed experiments, multivalent vaccines could prevent the promotion of the diseases caused by this bacterium and enhance the prevention chance of S. aureus infections.

A comparative study on the antimicrobial activity of irreversible hydrocolloid mixed with silver nanoparticles and chlorhexidine.

BACKGROUND: Impressions taken from patients have the potential of cross-transmission of infection among dentistry personnel. The present study aimed to compare the antimicrobial activity of chlorhexidine (CHX) and silver nanoparticles (AgNPs) combined with irreversible hydrocolloid. MATERIALS AND METHODS: This experimental study examined the in vitro antimicrobial effects of irreversible hydrocolloid mixed with silver nanoparticles and chlorhexidine using four groups, namely CHX (0.2%) solution and mouthwash mixed with irreversible hydrocolloid Groups 1 and 2), AgNPs (0.1 and 0.2%) (Groups 3 and 4), and specimens mixed with distilled water as a control group (Group 5) on bacterial strains, namely, Staphylococcu saureu s,Enterococcu sfaecali s,Pseudomona saeruginos a,Escherichi acoli, andStaphylococcu sepidermidi s through disc diffusion method. There were three replications per bacterial species. As data were not normally distributed, Kruskal-Wallis test was used at a significance level of 0.05. RESULTS: No antimicrobial activity was observed in the control groups. In S. aureus, CHX mouthwash had the highest antimicrobial activity, and AgNPs 0.1% and 0.2% groups had lower antimicrobial activity, and there was a significant difference between the two concentrations of AgNPs (P < 0.05). InE.faecali s, the effects of CHX compounds and AgNPs 0.2% were similar to each other and were higher than the effect of AgNPs 0.1% (P < 0.05). In E. coli, CHX compounds exhibited the highest efficacy relative to other materials (P < 0.05), and the AgNPs had no effect. In P. aeruginosa, AgNPs showed the highest growth inhibition zone, which had a significant difference compared to other materials (P ≤ 0.01), whereas the CHX compounds were not effective. InS.epidermidi s, the effect of CHX compounds was similar to one another and was higher than the effect of AgNPs (P ≤ 0.01). CONCLUSION: According to our observations, the antimicrobial activity of AgNPs at 0.1 and 0.2% against five tested bacterial strains was similar to those of pure CHX 0.2% solution and CHX 0.2% mouthwash.

Synthesis, characterization and assessment of anti-quorum sensing activity of copper(II)-ciprofloxacin complex against Pseudomonas aeruginosa PAO1.

Quorum sensing (QS) inhibition by metal-antibiotic complexes is a promising strategy for the management and control of multidrug resistant Pseudomonas aeruginosa infections. We investigated the anti-quorum sensing activity of sub-minimum inhibitory concentration (sub-MIC) of copper(II) sulfate pentahydrate-ciprofloxacin (Cu-CIP) complex and free ciprofloxacin (free-CIP) against P. aeruginosa PAO1. Copper-CIP complex was synthesized and its characterization was assessed using spectroscopic methods and single crystal X-ray analysis. The effect of sub-MIC (1/4 and 1/16 MIC) concentrations of Cu-CIP and free-CIP on cell growth, biofilm formation, motility, alginate and pyocyanin production, H2O2 susceptibility and expression of QS circuit genes lasI and lasR in PAO1 was determined. Minimum inhibitory concentration of Cu-CIP complex and free-CIP was determined as 0.125 µg/ml. Copper-CIP complex did not show significant effect on the cell growth at concentrations of 1/4 and 1/16 MIC. However, sub-MIC concentrations (1/4 and 1/16 MIC) of Cu-CIP showed the significant reduction in violacein production, motility, biofilm formation, alginate and pyocyanin production and sensitivity to H2O2 in a concentration dependent manner (P < 0.001). Copper-CIP at the concentration of 1/4 MIC showed the greatest reduction in lasI and lasR transcriptional expression (89.5% and 96.2% respectively). Considering the biological effects of Cu-CIP complex and its inhibitory activity on QS related virulence traits at low concentrations (0.03 and 0.007 µg/ml), it may be used as an effective approach in the management of infections caused by P. aeruginosa.

Inhibitory activity of metal-curcumin complexes on quorum sensing related virulence factors of Pseudomonas aeruginosa PAO1.

The use of metal complexes to reduce or inhibit virulence factors of Pseudomonas aeruginosa is a promising strategy for the management and control of infections caused by this multidrug-resistant pathogen. The present study aimed to investigate the anti-quorum sensing activity of sub-minimum inhibitory concentrations (sub-MIC) of copper(II) sulfate pentahydrate-curcumin complex (Cu-CUR), iron(III) nitrate nonahydrate -curcumin complex (Fe-CUR), zinc(II) chloride-curcumin complex (Zn-CUR) and free curcumin (free-CUR) against P. aeruginosa PAO1. Metal-CUR complexes were synthesized and characterized by spectroscopic methods. The effect of sub-MIC (1/4 and 1/16 MIC) concentrations of metal-CUR complexes and free-CUR on cell growth, biofilm formation, motility, alginate and pyocyanin production, H2O2 susceptibility and expression of lasI and lasR genes in PAO1 was determined. MIC of metal-CUR complexes and free-CUR was determined as 62.5 and 125 µg/ml, respectively. Metal-CUR complexes at concentration of 62.5 µg/ml significantly reduced the cell growth to 1.5%-3.3%. Although we did not measure the anti-QS activity of metal-CUR complexes directly against PAO1, they indicated anti-QS activity in C. violaceum CV026. Copper-CUR complex at the concentration of 1/4 MIC showed the greatest inhibitory effect on swarming and twitching motilities, biofilm formation, alginate and pyocyanin production, sensitivity to H2O2 and reduction in the expression levels of lasI and lasR genes (P < 0.001). Considering the biological effects of Cu-CUR complex and its inhibitory activity on virulence factors, it may be used as an effective compound for treatment and control of infections caused by P. aeruginosa.

Frequency of hemolysin BL and non-hemolytic enterotoxin complex genes of Bacillus cereus in raw and cooked meat samples in Zanjan, Iran.

Food safety has emerged as an important global issue with international trade and public health implications. Bacillus cereus is an important cause of food poisoning worldwide. A total of 200 individual meat samples were collected from meat retail outlets and restaurants and investigated the frequency of B. cereus and hemolysin BL (Hbl), non-hemolytic enterotoxin (Nhe) complex genes. The meat samples were immediately homogenized and cultured on Bacillus cereus selective agar and subjected for confirmatory biochemical tests and molecular detection of gyrB, hblA, hblC, hblD, nheA, nheB and nheC genes. A total of 29 (14.5 %) meat samples were positive for the presence of B. cereus. The frequency of B. cereus in raw meat (14.1 %) was similar to cooked beef samples (15 %) (P >  0.05). Twenty six (89.6 %) isolates carried at least one or more enterotoxin genes. We found nheA (58.6 %) and hblD (51.7 %) genes with higher frequency than others. Hemolysin BL complex genes were found in lower frequency than Nhe complex (P >  0.05). Detection of enterotoxigenic B. cereus in meat samples shows a probable risk for public health. Therefore, the reliable molecular methods for monitoring of potentially pathogenic B. cereus are strongly recommended for the routine food examination.

Molecular Characterization of Methicillin and Vancomycin Resistant Staphylococcus aureus Strains Isolated from Hospitalized Patients.

Objectives: Nosocomial infections caused by methicillin resistant Staphylococcus aureus (MRSA) and emergence of vancomycin resistant S. aureus (VRSA) have led to great concern in healthcare settings worldwide. Methods: A total of 100 S. aureus clinical isolates from hospitalized patients were investigated for antimicrobial susceptibility, the presence of resistance (mecA, vanA, and vanB) and virulence (hlaA, fnbpA, cna, clfA, tsst-1, eta, and spa) encoding genes, and molecular typing based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of spa gene. Results: All isolates were resistant to one or more antibiotics, with the most frequent resistance found against amoxicillin (69%). A total of 46 isolates were MRSA, and 40% of isolates were multidrug resistant (MDR). Of all S. aureus isolates, two isolates were confirmed as VRSA and four isolates confirmed as vancomycin intermediate S. aureus (VISA). The frequency of clfA, cna, tsst-1, and eta genes among MRSA isolates was significantly higher than methicillin sensitive S. aureus (MSSA). The significant correlation between MDR isolates and the carriage of multiple virulence genes was seen. All MDR isolates carried at least four virulence genes. Furthermore, biofilm formation in MRSA isolates was significantly higher than MSSA. The spa gene PCR products generated 4 major and 10 minor types. After digestion of spa amplicons with HindIII restriction enzyme, 10 different patterns ranging 174-938 bp were detected. S2b and S2a subtypes were detected frequently in MRSA isolates. Conclusions: It seems that the appropriate surveillance and control measures are essential to prevent the emergence and transmission of MRSA and VRSA strains in our country.

Resistance pattern and distribution of carbapenemase and antiseptic resistance genes among multidrug-resistant Acinetobacter baumannii isolated from intensive care unit patients.

PURPOSE: Nosocomial infections caused by multidrug resistant Acinetobacter baumannii have emerged as a serious problem in healthcare settings worldwide. METHODOLOGY: A total of 100 A. baumannii clinical isolates from immunocompromised patients hospitalized in ICUs in Iran were investigated for antimicrobial susceptibility and the presence of carbapenemase and antiseptic resistance genes. RESULTS: All isolates were resistant to one or more antibiotics, with the most frequent resistance found against ciprofloxacin and imipenem (100 %) and piperacillin (99 %). The MICs of biocides were determined by the agar dilution method. No apparent resistance to biocides was seen among the 100 A. baumannii isolates. All isolates were effectively inhibited by the user's defined concentrations of cetrimide, benzalkonium chloride and glutardaldehyde. The intrinsic ß-lactamase gene, blaOXA-51-like, was detected in all A. baumannii isolates. Coexistence of blaOXA-51 andblaOXA-23 was encountered in 89 % of isolates. However, genes blaOXA-58, blaSIM and blaIMP were not detected in any isolates. While A. baumannii isolates were sensitive to biocides, they carried qac genes with the qacEΔ1 gene being the most common, at a frequency of 91 %. CONCLUSION: Our study revealed the high frequency of multidrug- and carbapenem-resistant isolates of A. baumannii in ICU patients, with a high prevalence of the genes blaOXA-23 and blaOXA-51. However, no apparent biocide resistance was seen in A. baumannii isolates. It appears that appropriate surveillance and control measures are essential to prevent the emergence and transmission of MDR A. baumannii in Iran.