Vancomycin-resistant S. Aureus Isolates Research Articles

Phenotypic and genotypic characterization of methicillin and vancomycin-resistant Staphylococcus aureus isolated from human clinical samples in Kerman hospitals, Iran

Today, methicillin-resistant S. aureus (MRSA) isolates are widely spread in both communities and hospitals, causing high morbidity and mortality worldwide. However, the emergence of vancomycin resistance among MRSA isolates has been perceived as a formidable threat to therapeutic management. Therefore, the present study aimed to determine the frequency and antibiotic susceptibility of MRSA and vancomycin-resistant S. aureus (VRSA) strains in Kerman hospitals in Iran. A total of 455 human clinical samples were collected from Kerman hospitals. The samples were examined for the presence of S. aureus and the isolates were tested for their susceptibility to antibiotics by disk diffusion method. MRSA and VRSA isolates were detected by a combination of phenotypic and genotypic methods. Sixty-five coagulase-positive S. aureus strains were isolated, and the higher isolation rate was from wound samples (47.7%) compared with blood (18.5%), respiratory tract secretions (10.8%), urine (10.8%), body fluids (9.2%), abscess (1.5%), and cerebrospinal fluids (1.5%). The isolates exhibited high resistance toward tetracycline (69.2%). Thirty-four isolates (52.3%) were categorized as MRSA (phenotypically resistant to cefoxitin). mecA gene was amplified in 97.1% of MRSA strains. Moreover, only one MRSA strain was resistant to vancomycin (MIC=128), and this isolate carried the vanA gene. The frequency of MRSA was not significantly associated with gender, age, sample type, and sampling locality (p>0.05). Given the alarming rate of resistance among MRSA isolates, monitoring of antibiotic resistance should be performed to reduce treatment failure in patients with staphylococcal infections. Although vancomycin remains a drug of choice for MRSA, our study suggests that its efficacy may be limited by resistance development.

Methicillin-Resistant Staphylococcus aureus and Coagulase-Negative Staphylococcus from School Dining Rooms in Argentina.

Methicillin-resistant Staphylococcus aureus (MRSA) constitutes an important cause for concern in the field of public health, and the role of the food chain in the transmission of this pathogen and in antimicrobial resistance (AMR) has not yet been defined. The objectives of this work were to isolate and characterize coagulase-positive Staphylococcus (CoPS) and coagulase-negative Staphylococcus (CoNS), particularly S. aureus, from school dining rooms located in Argentina. From 95 samples that were obtained from handlers, inert surfaces, food, and air in 10 establishments, 30 Staphylococcus strains were isolated. Four isolates were S. aureus, and the remaining ones (N = 26) belonged to 11 coagulase-negative species (CoNS). The isolates were tested for susceptibility to nine antibiotics. The presence of genes encoding toxins (luk-PV, sea, seb, sec, sed, and see), adhesins (icaA, icaD), and genes that confer resistance to methicillin (mecA) and vancomycin (vanA) was investigated. The resistance rates measured for penicillin, cefoxitin, gentamicin, vancomycin, erythromycin, clindamycin, levofloxacin, trimethoprim-sulfamethoxazole, and tetracycline were 73%, 30%, 13%, 3%, 33%, 17%, 13%, 7%, and 7% of the isolates, respectively. Seventeen AMR profiles were detected, and 11 isolates were multidrug resistant (MDR). Seven methicillin-resistant Staphylococcus isolates were detected in the hands of handlers from four establishments, two of them were MRSA. Two S. aureus isolates presented icaA and icaD, another one, only icaD. The gene vanA was found in two isolates. In relation to S. aureus, resistance to vancomycin but not to gentamicin was detected. School feeding plays a key role in the nutrition of children, and the consumption of food contaminated with MRSA and vancomycin-resistant S. aureus (VRSA) can be a serious threat to health. In particular, it was detected that the handlers were the source of MRSA, VRSA, MR-CoNS (methicillin-resistant coagulase-negative Staphylococcus), and MDR isolates. The results obtained indicate that the vigilance of this pathogen in school dining rooms should be extreme.

Staphylococcus aureus Induced Wound Infections Which Antimicrobial Resistance, Methicillin- and Vancomycin-Resistant: Assessment of Emergence and Cross Sectional Study.

Wound infection is a prevalent concern in the medical field, being is a multi-step process involving several biological processes. Methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) infections often occur in areas of damaged skin, such as abrasions and open wounds. This research aims to light the incidence of MRSA and VRSA in wound swabs, the antimicrobial susceptibility configuration of isolated S. aureus patterns in pus/wound samples collected from Saudi Arabian tertiary hospital. The cross section study, β- lactamase detection, VRSA genotyping, MAR index, D-test and VRSA genotyping are methods, which used for completed this research. Patients of several ages and genders delivered specimens from two hospitals in the Al jouf area, in the northern province of Saudi Arabia. S. aureus was found in 188 (34.7%) of the 542 wounds. The traumatized wounds provided 71 isolates (38.8%), surgical wound provided 49 isolates (26.8%) and abscess were represented 16 by isolates (8.7%). In the study, 123 (65.4%) out of 188 were MRSA, 60 (31.9%) were MSSA, and five (2.7%) were VRSA. Linezolid and rifampin were found to be the most effective antimicrobials with 100% in vitro antibacterial activity against S. aureus isolates. The Multiple antimicrobials resistance (MAR) index revealed 73 isolates (38.9%) with a MAR index greater than 0.2, and 115 (61.1%) less than 0.2. The D-test showed that of MLSb phenotypes among S. aureus, 22 (11.7%) strains were D-test positive (MLSbi phenotype), 53 (28.2%) strains were constitutive MLSc phenotypes, and 17 (9%) strains were shown to have MSb phenotypes. All VRSA isolates (n=5) were found to be positive for vanA, and no vanB positive isolates were detected in the study. Regular monitoring and an antimicrobials stewardship program should be in place to provide critical information that can be utilized for empirical therapy and future prevention strategies.

Evidence and Molecular Characterization of Multidrug Resistant Staphylococcus aureus Isolated From Equines in Pakistan

The purpose of the current study was to investigate the nasal colonization and drug resistance profile of Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), and vancomycin-resistant S. aureus (VRSA) in donkeys (n =63), mules (n=42), and horses (n=98). MRSA and VRSA were confirmed based on phenotypic and molecular methods, followed by phylogenetic analysis. Furthermore, the association of various animal and management-based risk factors with S. aureus colonization was also evaluated. The presence of nuc gene on polymerase chain reaction showed an overall prevalence of 42.86% of S. aureus in equines. Based on Kirby-Bauer disc diffusion test, 26.44% of isolates showed resistance to vancomycin. Further, mecA and vanB genes were targeted which showed an overall 14.94% and 13.79% prevalence of methicillin and vancomycin-resistant isolates, respectively. The phylogenetic analysis revealed a significant variation of the study isolates with each other and with already reported sequences of mecA and vanB genes. Risk factor analysis revealed that raising purpose (P=.011), work intensity (P < .001), stocking density (P=.006), presence of other livestock animals in surroundings (P=.043), and common drinking water source (P=.023) as significant. Antimicrobial susceptibility testing of MRSA and VRSA isolates showed high resistance to various commonly used antibiotics. Furthermore, all the tested isolates showing resistance to three or more than three antibiotics were considered multiple drug-resistant. The current study manifests the molecular evidence of MRSA and VRSA isolated from equines in Pakistan which will help to address the emerging issue of multidrug resistance in S. aureus in equines and emphasizes the need for possible measures to tackle this issue.

Antibacterial and antibiofilm activities of taxifolin against vancomycin-resistant S. aureus (VRSA)

Background and objective: The medicinal effects of flavonoids are widely described in the literature; however, their antimicrobial effects against antibiotic resistant bacteria are yet to be highlighted. This study was aimed at investigating the growth and biofilm inhibitory effects of taxifolin, a flavonoid, against vancomycin-resistant Staphylococcus aureus (VRSA). Methods: Seven VRSA isolates were used to assess the antimicrobial and antibiofilm influence of taxifolin. The agar-well diffusion method was used to determine the zones of inhibition caused by taxifolin, and resazurin-based microdilution technique was used to assess the minimum inhibitory concentration. Crystal violet staining technique was used to assess the biomass of biofilms formed by the microorganisms. GraphPad Prism software was used to present the data in figures. Results: Taxifolin inhibited bacterial growth in a dose-dependent fashion and reduced bacterial viability. It similarly attenuated the biofilm production activity of bacterial isolates in a dose-dependent manner. Conclusions: Current findings suggest the antibacterial and antibiofilm influence of taxifolin against VRSA in a dose-dependent manner.

Genotyping of enterotoxigenic methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Staphylococcus aureus (VRSA) among commensal rodents in North Sinai, Egypt.

This study aimed to identify genotype enterotoxigenic antimicrobial-resistant Staphylococcus aureus species, mainly methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) among commensal rodents. A total of 280 samples were collected from nasal and mouth swabs, heart blood, intestinal content and lung tissues of 56 commensal rodents trapped from North Sinai, Egypt. Antimicrobial susceptibility testing was performed to bacteriologically identified S. aureus isolates against 15 antimicrobial agents by disc diffusion method. Detection was conducted for identifying coagulase gene (coA), antimicrobial-resistant genes (mecA and vanA/B), enterotoxigenic and virulence determinant genes (hlg, seb, sed and see) among the MRSA and VRSA isolates. Staphylococcus aureus species were isolated from 24 (42.86%) out of 56 rodents. Phenotypic examination revealed that all the isolates were multidrug-resistant, whereas two isolates were multiple antibiotic resistant (MAR). Out of 33 examined isolates, 33 (100%) were resistant to oxacillin and amoxicillin, 31 (93.93%) to cefoxitin and 12 (36.36%) to vancomycin. PCR assay revealed that 24 isolates revealed (100%) positivity to coA gene, 17 (70.83%) to mecA gene and 12 (50%) to vanA/B genes. Enterotoxin genes and haemolysin genes were detected among MRSA and VRSA isolates. There was a strong positive correlation between the tested antimicrobial-resistant genes and virulence genes (p>0.05). This study demonstrated the occurrence of MRSA and VRSA strains among commensal rodents in North Sinai, Egypt. The detection of enterotoxigenic and virulence genes of the isolated MRSA and VRSA strains indicated the health hazards of food contamination and zoonotic infections. This study emphasizes the role of commensal rodents in maintaining and disseminating multidrug-resistant MRSA and VRSA strains to the environment, animals and human beings.

Molecular Confirmation of Vancomycin-Resistant Staphylococcus aureus with vanA Gene from a Hospital in Kathmandu.

Staphylococcus aureus, a commensal on the skin and in the nasal cavity of humans, is one of the most serious cases of nosocomial infections. Moreover, methicillin-resistant S. aureus (MRSA) is a leading cause of morbidity and mortality worldwide. For the treatment of MRSA infections, vancomycin is considered as a drug of choice. However, the emergence of vancomycin resistance among MRSA isolates has been perceived as a formidable threat in therapeutic management. To estimate the rate of vancomycin-resistant S. aureus (VRSA) and to detect the vancomycin-resistant genes, namely, vanA and vanB, among the isolates, a hospital-based cross-sectional study was conducted from July to December 2018 in Annapurna Neurological Institute and Allied Science, Kathmandu, Nepal. S. aureus was isolated and identified from different clinical samples and processed for antibiotic susceptibility testing by the modified Kirby–Bauer disc diffusion method. The screening of MRSA was performed as per Clinical and Laboratory Standard Institute (CLSI) guidelines. VRSA was confirmed by the minimum inhibitory concentration (MIC) method by employing E-test strips. All the phenotypically confirmed VRSA were further processed to detect the vanA and vanB gene by using the conventional polymerase chain reaction (PCR) method. A total of 74 (20.3%) S. aureus were isolated, and the highest percentage of S. aureus was from the wound samples (36.5%). Of 74 S. aureus isolates, the highest number (89.2%) was resistant to penicillin, and on the other hand, linezolid was found to be an effective drug. Likewise, 45 (60.81%) were found to be MRSA, five (11.11%) were VRSA, and 93.2% of S. aureus isolates showed an MAR index greater than 0.2. Two VRSA isolates (40%) were positive for the vanA gene. The higher prevalence of MRSA and significant rate of VRSA in this study recommend routine surveillance for the MRSA and VRSA in hospital settings before empirical therapy.

Isolation and Genetic Characterization of Vancomycin-resistant and mecC+ Methicillin-resistant Staphylococcus aureus Strains in Clinical Samples of Bojnurd, Northeastern Iran

Background: The emergence of antibiotic-resistant Staphylococcus aureus strains is one of the major concerns about the various staphylococcal infections. Vancomycin is one the most important effective antibiotics on staphylococcal lethal infections. To date, vancomycin-resistant strains are increasingly isolated in different parts of the world, and it is alerting. Objectives: The current study was designed to evaluate the prevalence, and antibiotic susceptibility pattern of methicillin-resistant S. aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) isolates in the main tertiary hospital of Bojnurd, Iran. Methods: Staphylococcus aureus isolates were collected from different clinical samples in Imam Reza Hospital of Bojnurd. After identification of isolates through using conventional methods, they were evaluated by agar screening, disk diffusion, and minimum inhibitory concentration (MIC) methods to determine resistance to vancomycin and methicillin. We also performed polymerase chain reaction (PCR) for the detection of mecA, mecC, vanA, and vanB genes. After confirmation of vancomycin resistance, genetic analysis was performed using SCCmec, agr, and spa typing, and multilocus sequence typing (MLST) methods on VRSA isolates. Results: We found four vancomycin-resistant isolates (1.29%). Also, 75% of isolates were resistant to cefoxitin. Using the PCR method, mecA was found in 73%, mecC in 0.64%, and vanA in 1.29% of isolates. Interestingly, we found two mecC positive isolates in MRSA isolates. The alpha-hemolysin (81.81%) and enterotoxin C (27%) had the highest and lowest toxins percentage, respectively. Among mecA positive isolates, SCCmec IV (37%), SCCmec III (31.27%), SCCmec I (14%), SCCmec II (11%), and SCCmec V (5.7%) were the most prevalent SCCmec types, respectively. It should be noted that the two mecC positive isolates belonged to SCCmec XI. Agr I (76.29%) was the highest agr type. We recognized t037 as the dominant spa type, and ST239, ST6, ST97, and ST8 were found in VRSA isolates. Conclusions: In our study, the frequency of mecA genes in MRSA isolates was very high. It seems that the resistant isolates belonged to endemic clones of Iran.

Vancomycin resistant Staphylococcus aureus from Clinical Isolates in Benin City, Nigeria

Multidrug resistant Staphylococcus aureus has increasingly been implicated as an agent of clinical infections worldwide. This study investigated the prevalence of Vancomycin resistant S. aureus (VRSA) among S. aureus causing infections in Benin City, Nigeria. A total of 400 non-repetitive S. aureus isolates were obtained from clinical specimens in the University of Benin Teaching Hospital. These isolates were identified by standard microbiological tests. Antimicrobial susceptibility tests and screening for vancomycin resistance (using phenotypic method) was carried out for all isolates. Eighteen isolates (4.5%) were resistant to vancomycin with 10 (2.5%) VRSA isolates recovered from females while 8(2.0%) were isolated from males (p = 0.9981). Isolates from catheter tip had the highest prevalence of VRSA (33.3%) and showed statistical significance in comparison with other clinical specimens (p = 0.0036). Vancomycin resistant S. aureus isolates showed poor susceptibility to commonly used antibacterial agents (&lt; 50%). In order to mitigate the effect of selective pressure on the survival and proliferation of resistant strains, prudent use of vancomycin and other antimicrobial agents is advocated. The need for continuous surveillance, susceptibility testing and screening cannot be overemphasized.

English

The increase in resistance pattern of Staphylococcus aureus (S. aureus) has been an emerging threat in therapeutic areas of the dairy industry throughout the globe. The current study was conducted in bovines of district Faisalabad, Pakistan to investigate the phenotypic prevalence of methicillin-resistant (MRSA) and vancomycin-resistant S. aureus (VRSA) in milk samples positive for subclinical mastitis. The study further aimed to assess the associated risk factors and antimicrobial susceptibility pattern against MRSA and VRSA isolates. A total of 385 milk samples (n=193 cattle; n=192 buffalo) collected and screened for subclinical mastitis by surf field mastitis test (SFMT) were further subjected to standard microbiological techniques for the isolation of S. aureus. The positive isolates of S. aureus were phenotypically evaluated for MRSA and VRSA by the disc diffusion method. The study results revealed that out of 385 milk samples, 45.97% (177/385) samples were found positive for subclinical mastitis on SFMT while 37.14% (143/385) samples were confirmed for the presence of S. aureus. Out of these S. aureus isolates, MRSA and VRSA were confirmed in 17.48% (25/143) and 12.58% (18/143) samples respectively. The in-vitro trials of various antibiotics for MRSA and VRSA isolates showed 100% resistance towards Cefoxitin followed by 50% towards Gentamicin, Tylosin, and Trimethoprim + Sulfamethoxazole then 25% to Oxytetracycline, and Fusidic acid while ciprofloxacin, moxifloxacin, and linezolid were found sensitive against study isolates. The public health importance of S. aureus and emerging resistance against antibiotics like methicillin and vancomycin demands regular monitoring of effective use of antimicrobial agents against the isolates of VRSA and MRSA.

Effect of silver nanoparticles on vancomycin resistant Staphylococcus aureus infection in critically ill patients

ABSTRACT A prevalent increase in antimicrobial resistance represents a universal obstacle for the treatment of Staphylococcus aureus (S. aureus) infection, especially in critically ill patients. Silver nanoparticles are defined as broad spectrum bactericidal agents, which might be effective against vancomycin resistant S. aureus (VRSA). In this study, we examined the bactericidal efficacy of silver nanoparticles on VRSA in 150 blood and sputum samples isolated from intensive care patients. Methicillin resistant S. aureus (MRSA) isolates were identified in 83 samples, with an incidence of 55.3%. Meanwhile, VRSA isolates were found in 11 and 8 isolates (a total of 19 isolates out of 150) from sputum and blood samples, with an incidence of 14.67% and 10.67%, respectively, with a total incidence of 12.67%. Vancomycin intermediate S. aureus (VISA) isolates had an inhibitory zone ranging from 9 to 13 mm, which was found in 13 out of 19 isolates, whereas VRSA isolates had an inhibitory zone ranging from 0 to 6 mm, which was detected in 6 out of 19 isolates. The findings of this study confirm that silver nanoparticles are an effective treatment against VRSA.

Identification and evaluation of Anisochilus carnosus (L. fil.) Wall. as a novel candidate with therapeutic potential against multi-drug resistant Staphylococcus aureus

BackgroundStaphylococcus aureus has been constantly emerging with newer drug resistance patterns; vancomycin intermediate S. aureus (VISA) and vancomycin resistant S. aureus (VRSA) being recent entries. Hence the growing quest for newer antibiotics has entered the realm of the plants as they are less explored and usually have negligible side effects. Although a number of medicinal plants are being used traditionally, their spectrum and mechanism of action are not completely deciphered. ObjectivesTo evaluate the aqueous (the traditionally administered extract) and acetone/ethanol (organic solvents with the highest capacity to extract phytoconstituents) extracts of ten medicinal plants traditionally used to treat respiratory infections in south India for antibacterial activity against S. aureus. MethodsThe antibacterial activity against S. aureus standard strain was evaluated by the broth dilution method. The extracts exhibiting antibacterial activity were further validated using 50 S. aureus isolates by the broth dilution and agar diffusion methods. ResultsOnly Anisochilus carnosus acetone extract (ACace; MIC-0.33 ± 0.14 mg/mL) exhibited significant antibacterial activity, and its zone diameter break points were determined. The sensitivity of the 50 S. aureus clinical isolates to ACace (72 %) was comparable to methicillin (68 %), linezolid (94 %) and clindamycin (82 %). Additionally, ACace, inhibited majority of the MRSA (87.5 %), VISA (70 %) and VRSA (83 %) isolates. ConclusionThis is the first study to report the antibacterial potential of a novel candidate, ACace against S. aureus, evaluate its inhibitory effect on clinical isolates including drug-resistant strains and highlight the importance of zone diameter break points for the evaluation of medicinal plant extracts.

The prevalence of vanA gene in clinical isolates of vancomycin-resistant Staphylococcus aureus in a hospital in Mazandaran, Iran

Staphylococcus aureus is one of the most important causes of infections in hospitals. Although vancomycin is often prescribed for the treatment of methicillin-resistant S. aureus (MRSA), vancomycin-resistant S. aureus (VRSA) isolates remain a major problem in hospitals. In this study, we investigated the prevalence of vancomycin-resistant S. aureus isolates and also vanA gene in these clinical isolates in a hospital in Mazandaran Province, Iran. In this cross-sectional study, a total number of 60 S. aureus samples were identified from different clinical specimens after standard biochemical and microbiological tests. Disk agar diffusion test using Kirby-Bauer method was applied for antibiotics against S. aureus isolates. The presence of vanA gene was investigated in VRSA and intermediate resistance (VISA) isolates by PCR method using specific primers. Over 40% of isolates were resistant to commonly-used antibiotics, including gentamicin (46.67%), ceftazidime (45%) and carbenicillin (43.34%). Only few, however, were sensitive to gentamicin (33.33%) and ceftazidime (35%). Vancomycin was the most effective antibiotic against S. aureus isolates (56.66% sensitivity). Eleven isolates (18.34%) were resistant (VRSA) and 15 isolates (25%) were intermediate resistance (VISA) to vancomycin. Molecular analysis of vanA gene in 11 VRSA and 15 VISA showed that 8 VRSA (72.72%) and 6 VISA (40%) isolates were positive for vanA gene. The incidence of VRSA and VISA strains, as well as the frequency of vanA gene in these isolates are high and emerging in Mazandaran hospitals. There is a need to keep the emergence and spread of these strains to a minimum level.

Vancomycin resistance among Staphylococcus aureus isolates in a rural setting, Egypt.

With the increased occurrence of methicillin resistant S. aureus (MRSA), the consumption of vancomycin, the drug of choice, has also increased. As a consequence, strains of S. aureus resistant to vancomycin have started to emerge. This study aimed to evaluate the level of vancomycin resistance among clinical and nasal S. aureus isolates in a rural town in Egypt. This cross-sectional study was held in the general hospital at the rural town of Kafr Eldawar in Egypt, during the period from January 2013 to January 2014. S. aureus isolates were collected from clinical samples and from nasal swabs. Two hundred S. aureus isolates were collected, 80 (40%) from clinical samples and 120 (60%) from nasal carriage samples. Vancomycin resistant S. aureus (VRSA) was only detected in clinical samples, all collected from the outpatient clinic. Eleven VRSA isolates (13.8% of total S. aureus clinical isolates) and one strain of vancomycin-intermediate S. aureus (from nasal carriage) were detected. VRSA isolates were most resistant to ciprofloxacin (90.9%) and erythromycin (81.8%). Five isolates were resistant to all tested antibiotics: ciprofloxacin, clindamycin, erythromycin, linezolid, oxacillin, penicillin and trimethoprim-sulfamethoxazole. MRSA was found to constitute 43.8% of clinical S. aureus isolates. The MRSA colonization rate among community individuals was 43.6%, 42.9% among healthcare workers and 51.4% among patients. The prevalence of VRSA was high in clinical samples suggesting that there is a high level of VRSA strains in Egypt that goes undetected since most laboratories only use disk diffusion for detection of vancomycin resistance.

Molecular characterization of vancomycin-resistant Staphylococcus aureus strains isolated from clinical samples: A three year study in Tehran, Iran.

IntroductionEmergence of vancomycin-intermediate Staphylococcus aureus (VISA) and vancomycin-resistant S. aureus (VRSA) strains has led to great concern in global public health in both developing and developed countries. This study investigated distribution and molecular characterization of VRSA strains in Tehran’s hospitals using a combination of molecular typing methods.Materials and methodsA total of 1789 S. aureus isolates obtained between 2014 and 2017 and were characterized using antibiogram, SCCmec typing, spa typing, and multilocus-sequence typing. Resistance to vancomycin was determined by E-test method. After confirmation of the isolated VRSA strain, genetic analysis was performed by evaluating vanA and vanB genes presence.The presence of resistance (ermA, ermB, ermC, mupA, msrA, msrB, tetM, ant (4΄)-Ia, aac (6΄)-Ie/aph (2˝), aph (3΄)-IIIa) and toxin (etb, eta, pvl, tst) encoding genes was investigated by the polymerase chain reaction (PCR) technique.ResultsOf all S. aureus tested isolates, four isolates were confirmed as VRSA isolates and two isolates confirmed as VISA isolates. ST5- SCCmec II/t002 and ST239-SCCmec III/t037 strains had MIC values of 512μg/ml, ST239-SCCmec III/t037 and ST8-SCCmecIV/t008 strains had MIC values of 64μg/ml and ST22-SCCmec IV/t790 and ST239-SCCmec III/t030 strains had MIC values ≥ 8 μg/ml. pvl-encoding gene was confirmed in ST8-SCCmecIV/t008 and ST22-SCCmec IV/t790 strains. The isolates differed in the carriage of resistance and toxin encoding genes.ConclusionsThe study revealed the existence of VRSA strains in capital of Iran, Tehran. To our knowledge, this is the first report of ST239-SCCmec III/t037 as VRSA strain. These findings support the need for future surveillance studies on VRSA strains to keep the emergence and transmission of these isolates to a minimum.

Genomic analysis of vancomycin-resistant Staphylococcus aureus VRS3b and its comparison with other VRSA isolates.

High-level vancomycin resistance among Staphylococcus aureus poses a grave threat to global health as the treatment options for this pathogen are very limited. A detailed evaluation of the genetic background of vancomycin-resistant S. aureus (VRSA) is expected to facilitate the understanding of its origin and pathogenicity. In this study, we performed the genetic analysis of the clinical VRSA isolates and identified the genetic basis of resistance to multiple antibiotics among these strains, based on the available draft genome sequences. In addition, we generated the draft genome of the strain VRS3b, which was considered to be same as VRS3a based on its isolation from the same patient. We found that strain VRS3b did not harbor the genes responsible for tetracycline and gentamicin, which was further confirmed by the sensitivity towards these antibiotics. Our results suggest that the strains VRS3a and VRS3b are different from the view of antibiotic resistance and highlight the possibility of generation of two distinct VRSA strains from the same patient.

Characterization of SCCmec and spa types of methicillin-resistant Staphylococcus aureus isolates from health-care and community-acquired infections in Kerman, Iran

Spread of methicillin-resistant Staphylococcus aureus (MRSA) isolates is a worldwide problem. Molecular typing is a useful tool to understand MRSA epidemiology. Herein, we determined vancomycin-resistant, SCCmec and spa types among MRSA isolates recovered from healthcare and community-acquired infections in Kerman, Iran. A total of 170 S. aureus isolates were collected from different patients who were admitted to affiliated hospitals of Kerman University of Medical science. MRSA and vancomycin-resistant S. aureus (VRSA) isolates were detected by phenotypic methods. Polymerase chain reaction (PCR) technique was used for detection of mecA, vanA and vanB genes. Staphylococcal cassette chromosomemec (SCCmec) and spa typing were used for molecular typing of among MRSA isolates. Overall, 53% of isolates were considered as MRSA. Two MRSA isolates were resistant to vancomycin and vanA was detected in only one of VRSA isolates. SCCmec type III belonged to spa types t030 and t459 which they were the dominant spa types among community-associated MRSA (CA-MRSA) and healthcare-acquired MRSA (HA-MRSA) isolates. Our findings showed that the SCCmec type I and III spread from hospital settings to community, although the SCCmec type IV spread from community to healthcare systems. We have also reported VRSA isolates from hospitalized patients, therefore, appropriate policies should be enforced in order to prevent the spread of antibiotic resistance isolates in hospitals settings.

Vancomycin resistance among methicillin resistant Staphylococcus aureus isolates from intensive care units of tertiary care hospitals in Hyderabad

Background & objectives:Multidrug resistant methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of nosocomial and community acquired infections and is on the rise. The glycopeptide vancomycin has been proposed as the drug of choice for treating such infections. The present study aimed at identifying the vancomycin resistance both phenotypically and genotypically among the MRSA isolates from two tertiary care hospitals in Hyderabad, south India.Methods:MRSA were isolated and identified from different clinical samples collected from ICUs of tertiary care hospitals in Hyderabad using conventional methods. Antibiogram of the isolates and vancomycin MIC were determined following CLSI guidelines. vanA was amplified by PCR using standard primers.Results:All vancomycin resistant S. aureus (VRSA) isolates were MRSA. The VRSA isolates were positive for vanA gene, except one which was negative. All VRSA had a vancomycin MIC in the range of 16-64 mg/l.Interpretation & conclusions:The increase in vancomycin resistance among MRSA and excessive use of antimicrobial agents have worsened the sensitivity. Larger studies need to be done in various geographical regions of the country to better define the epidemiology, mechanism of vancomycin resistance in S. aureus and its clinical implications.

In vitro activity of oritavancin against community-associated meticillin-resistant Staphylococcus aureus (CA-MRSA), vancomycin-intermediate S. aureus (VISA), vancomycin-resistant S. aureus (VRSA) and daptomycin-non-susceptible S. aureus (DNSSA)

Isolates of community-associated meticillin-resistant Staphylococcus aureus (CA-MRSA), vancomycin-intermediate S. aureus (VISA), vancomycin-resistant S. aureus (VRSA) and daptomycin non-susceptible S. aureus (DNSSA) are increasing in frequency and new antistaphylococcal therapies are needed. Microdilution testing using Mueller–Hinton broth was used to determine the minimal inhibitory concentrations (MICs) of oritavancin and nine additional antimicrobial agents against 92 CA-MRSA, 23 VISA, 7 DNSSA and 10 VRSA isolates. Minimal bactericidal concentrations were also determined. Pulsed-field gel electrophoresis (PFGE) was performed. Staphylococcal cassette chromosome mec (SCC mec) typing as well as assays for Panton–Valentine leukocidin (PVL) and arginine catabolic mobile element (ACME) genes were performed. Oritavancin was more bactericidal than any of the other comparators against CA-MRSA and demonstrated excellent activity against VRSA and VISA.

Antimicrobial activities of ceragenins against clinical isolates of resistant Staphylococcus aureus.

The rise in the rates of glycopeptide resistance among Staphylococcus aureus isolates is concerning and underscores the need for the development of novel potent compounds. Ceragenins CSA-8 and CSA-13, cationic steroid molecules that mimic endogenous antimicrobial peptides, have previously been demonstrated to possess broad-spectrum activities against multidrug-resistant bacteria. We examined the activities of CSA-8 and CSA-13 against clinical isolates of vancomycin-intermediate S. aureus (VISA), heterogeneous vancomycin-intermediate S. aureus (hVISA), as well as vancomycin-resistant S. aureus (VRSA) and compared them to those of daptomycin, linezolid, and vancomycin by susceptibility testing and killing curve analysis. We also examined CSA-13 for its concentration-dependent activity, inoculum effect, postantibiotic effect (PAE), and synergy in combination with various antimicrobials. Overall, the MICs and minimal bactericidal concentrations of CSA-13 were fourfold lower than those of CSA-8. Time-kill curve analysis of the VRSA, VISA, and hVISA clinical isolates demonstrated concentration-dependent bactericidal killing. An inoculum effect was also observed when a higher starting bacterial density was used, with the time required to achieve 99.9% killing reaching 1 h with a 6-log10-CFU/ml starting inoculum, whereas it was>or=24 h with a 8- to 9-log10-CFU/ml starting inoculum with 10x the MIC (P<or=0.001). A concentration-dependent PAE was demonstrated with CSA-13, nearly doubling from 2x to 4x the MIC (P=0.03). With respect to the CSA-13 antimicrobial combinations, time-kill curve analysis showed no difference in the log10 CFU/ml at 24 h for the majority of the organisms tested. However, early synergy at 4 to 8 h was detected against the VRSA Pennsylvania strain (2002) when CSA-13 was tested in combination with gentamicin, while early additivity was demonstrated against all of the other organisms.