Agr Type Research Articles

Molecular Characterization, Virulence Determinants, and Antimicrobial Resistance Profile of Methicillin-Resistant Staphylococcus aureus in the North of Iran; a High Prevalence of ST239-SCCmec III/t037 Clone

Objectives: Emergence and prevalence of methicillin-resistant Staphylococcus aureus (MRSA) have become a major universal health concern, limiting therapeutic options. Methods: A total number of 37 MRSA isolates, including 19 clinical isolates from hospitalized patients and 18 colonizing isolates from health care workers were identified from 3 hospitals, in Gorgan, North of Iran. Antimicrobial susceptibility test was performed using the disk diffusion method and E-test. The presence of virulence and antibiotic resistance determinants were evaluated by PCR. The genotypical characterization was further analyzed using multi-locus sequence, spa, staphylococcal cassette chromosome, mec (SCCmec), and agr typing. Results: The frequency of MRSA among S. aureus isolates was 38.14% (37/97). The most frequent S. aureus resistant isolates were found to be obstinate against penicillin (98%) and gentamicin (82.5%). Additionally, the lowest resistance rates were found against daptomycin (0%), vancomycin (2.7%), and quinupristin-dalfopristin (5.4%). All MRSA isolates were susceptible to daptomycin with minimum inhibitory concentration (MIC)50/MIC90 of 0.25/0.5 μg/mL. One isolate belonging to sequence type 239 (ST239)-SCCmecIII/t037 clone (MIC ≥16 μg/mL) was resistant to vancomycin. All but 1 isolate that shares ST22-SCCmec IV/t790 strain were positive for both tsst and pvl genes. The most predominant MRSA isolates (27%) were associated with ST239-SCCmec III/t037, and ST239-SCCmec III/t924 (16.2%) clones, subsequently. In our study, circulating MRSA strains were genetically diverse with a high prevalence of ST239-SCCmec III/t037 clone. Conclusion: These findings emphasize the need for future and continuous surveillance studies on MRSA to prevent the dissemination of existing multidrug resistance MRSA clones in an effective manner.

Identification and detection of pathogenic bacteria from patients with hospital-acquired pneumonia in southwestern Iran; evaluation of biofilm production and molecular typing of bacterial isolates

BackgroundHospital-acquired pneumonia (HAP) is the second most common nosocomial infection in intensive care units (ICUs). The present study aims to determine the prevalence of pathogenic bacteria, their biofilm formation, and molecular typing from patients with HAP in southwestern Iran.MethodsFifty-eight patients with HAP participated in this cross-sectional study. Sputum and endotracheal aspirate were collected from each patient for isolation and detection of bacteria. Biofilm formation was evaluated using Congo red agar or Microtiter plate assay. The antimicrobial susceptibility patterns of the isolates were investigated. The multiplex polymerase chain reaction (M-PCR) technique was used to determine the Staphylococcal Cassette Chromosome mec (SCCmec) types of methicillin-resistant Staphylococcus aureus (MRSA) strains. All S. aureus isolates were typed using the agr typing method. A repetitive element sequence-based PCR (rep-PCR) typing method was used for typing of Gram-negative bacteria. Data were analyzed using the Statistical Package for the Social Sciences (SPSS) software version 15 and the chi-square test.ResultsBacteria were isolated in 52 (89.7%) of patients. Acinetobacter baumannii (A. baumannii) was the most prevalent organism (37%), followed by S. aureus, Pseudomonas aeruginosa (P. aeruginosa), and Escherichia coli (E. coli). Using the PCR method, 56 bacteria were detected. A. baumannii was the most prevalent (35.7%) organism. A. baumannii and P. aeruginosa were biofilm-producing. All Gram-negative isolates were colistin-sensitive, and most of the A. baumannii isolates were multidrug-resistant (MDR). MRSA was identified in 12 (80%) S. aureus isolates, and 91.6% of MRSA were SCCmec type III. The agr type III was the most predominant. The rep-PCR analysis showed seven different patterns in 20 A. baumannii, six patterns in 13 P. aeruginosa, and four patterns in 6 E. coli.ConclusionA. baumannii was more prevalent than S. aureus in ventilator-associated pneumonia (VAP), while S. aureus is a major pathogen in non-ventilator hospital-acquired pneumonia (NV-HAP), possibly due to the tendency of the former to aquatic environments. Based on the rep-PCR typing method, it was concluded that bacteria were transmitted from patients or healthcare workers among different wards. Colistin can be used as a treatment in Gram-negative MDR isolates.

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.

Clinical and Microbiological Profile of Diabetic Foot Ulcers Infected With Staphylococcus aureus in a Regional General Hospital in Bahia, Brazil.

It is necessary to know the resistance profile of Staphylococcus aureus to better control diabetic foot ulcer infections, to establish rational antibiotic therapy, and to avoid the development of resistant strains. This cross-sectional study evaluated the clinical parameters, virulence, and antimicrobial resistance profiles of S aureus in patients with diabetic foot disease admitted to a public hospital. S aureus strains were identified in patients with diabetes with amputation indication. Infected tissue samples were collected, microbes were isolated and identified. The microbial resistance profile was determined. Samples were also analyzed for biofilm formation and other virulence markers. The 34 individuals examined were mostly men, black, aged 60 years on average, and generally had a low income and education level. Most individuals had type 2 diabetes, and the mean time since diagnosis was 13.9 years. On an SF-36 (the Medical Outcomes Study 36-item short-form health survey) quality-of-life questionnaire, 75% of individuals obtained a score equal to 0 for physical impairment. S aureus specimens from 17 patients were isolated, corresponding to 50% of samples. Five isolates were classified as methicillin-resistant S aureus (MRSA). Molecular typing revealed that 20% of MRSA strains were SCCmec type V and 80% were type I. All isolates were sensitive to doxycycline; 61.5% were resistant to erythromycin, 38.5% to cefoxitin, 30.7% to clindamycin and ciprofloxacin, 23% to meropenem, 15.3% to gentamicin, 38.5% to oxacillin, and 7.7% (one strain) to vancomycin. Regarding biofilm production, 53% of samples were able to produce biofilms, and 84.6% had icaA and/or icaD genes. Additionally, the following enterotoxin genes were identified in the isolates: seb, sec, seg, and sei (5.9%, 5.9%, 11.8%, and 23.9%, respectively) and agr types 1 (5.9%) and 2 (11.8%). Genotypic evaluation made it possible to understand the pathogenicity of S aureus strains isolated from the diabetic foot; laboratory tests can assist in the monitoring of patients with systemic involvement.

Molecular Characterization of Staphylococcus aureus Isolated from Human and Food Samples in Northern Algeria.

Staphylococcus aureus is a commensal resident of the skin and nasal cavities of humans and can cause various infections. Some toxigenic strains can contaminate food matrices and cause foodborne intoxications. The present study aimed to provide relevant information (clonal complex lineages, agr types, virulence and antimicrobial resistance-associated genes) based on DNA microarray analyses as well as the origins and dissemination of several circulating clones of 60 Staphylococcus aureus isolated from food matrices (n = 24), clinical samples (n = 20), and nasal carriers (n = 16) in northern Algeria. Staphylococcus aureus were genotyped into 14 different clonal complexes. Out of 60 S. aureus, 13 and 10 isolates belonged to CC1-MSSA and CC97-MSSA, respectively. The CC 80-MRSA-IV was the predominant S. aureus strain in clinical isolates. The accessory gene regulator allele agr group III was mainly found among clinical isolates (70.4%). Panton–Valentine leukocidin genes lukF/lukS-PV were detected in 13.3% of isolates that all belonged to CC80-MRSA. The lukF/S-hlg, hlgA, and hla genes encoding for hemolysins and leucocidin components were detected in all Staphylococcus aureus isolates. Clinical and food isolates harbored more often the antibiotic resistance genes markers. Seventeen (28.3%) methicillin-resistant Staphylococcus aureus carrying the mecA gene localized on a SCCmec type IV element were identified. The penicillinase operon (blaZ/I/R) was found in 71.7% (43/60) of isolates. Food isolates belonging to CC97-MSSA carried several antibiotic resistance genes (blaZ, ermB, aphA3, sat, tetM, and tetK). The results of this study showed that all clones were found in their typical host, but interestingly, some nasal carriers had isolates assigned to CC705 thought to be absent in humans. The detection of MRSA strains among food isolates should be considered as a potential public health risk. Therefore, controlling the antibiotics prescription for a rational use in human and animal infections is mandatory.

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

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

Agr Typing and Detection of Mupirocin Resistance in Multi-Drug Resistant Clinical Isolates of Staphylococcus aureus in Northern Iran

Agr Typing and Detection of Mupirocin Resistance in Multi-Drug Resistant Clinical Isolates of Staphylococcus aureus in Northern Iran

Regulation of virulence and \u03b2-lactamase gene expression in Staphylococcus aureus isolates: cooperation of two-component systems in bloodstream superbugs

BackgroundMethicillin-resistant Staphylococcus aureus (MRSA)-bloodstream infections (BSI) are predominantly seen in the hospital or healthcare-associated host. Nevertheless, the interactions of virulence factor (VFs) regulators and β-lactam resistance in MRSA-BSI are unclear. This study aims to characterize the molecular relationship of two-component systems of VFs and the expression of the β-lactamase gene in MRSA-BSI isolates. In this study, 639 samples were collected from BSI and identified by phenotypic methods. We performed extensive molecular characterization, including SCCmec type, agr type, VFs gene profiles determinations, and MLST on isolates. Also, a quantitative real-time PCR (q-RT PCR) assay was developed for identifying the gene expressions.ResultsNinety-one (91) S. aureus and 61 MRSA (67.0%) strains were detected in BSI samples. The presence of VFs and SCCmec genes in MRSA isolates were as follows: tst (31.4%), etA (18.0%), etB (8.19%), lukS-PVL (31.4%), lukF-PV (18.0%), lukE-lukD (16.3%), edin (3.2%), hla (16.3%), hlb (18.0%), hld (14.7%), hlg (22.9%), SCCmecI (16.3%), SCCmecII (22.9%), SCCmecIII (36.0%), SCCmecIV (21.3%), and SCCmecV (16.3%). Quantitative real-time PCR showed overexpression of mecRI and mecI in the toxigenic isolates. Moreover, RNAIII and sarA genes were the highest expressions of MRSA strains. The multi-locus sequence typing data confirmed a high prevalence of CC5, CC8, and CC30. However, ST30, ST22, and ST5 were the most prevalent in the resistant and toxigenic strains.ConclusionWe demonstrated that although regulation of β-lactamase gene expressions is a significant contributor to resistance development, two-component systems also influence antibiotic resistance development in MRSA-BSI isolates. This indicates that resistant strains might have pathogenic potential. We also confirmed that some MLST types are more successful colonizers with a potential for MRSA-BSI.

Antimicrobial Resistance and Genetic Lineages of Staphylococcus aureus from Wild Rodents: First Report of mecC-Positive Methicillin-Resistant S. aureus (MRSA) in Portugal.

Simple SummaryStaphylococcus aureus is present in the microbiota of both humans and some animal species, being recognized as one of the most important opportunistic human pathogens. S. aureus is responsible for causing a variety of infections. Methicillin-resistant S. aureus (MRSA) is particularly important, as it is becoming increasingly prevalent in the population. MRSA has been increasingly reported among wild free-living animals which may impose a public health concern due to its zoonotic potential. To investigate the prevalence and antimicrobial resistance of S. aureus and MRSA in wild synanthropic rodent populations, we conducted this study on 204 rodents captured in port areas in Portugal. The antimicrobial resistance was investigated in all isolates as well as virulence genes and genetic lineages. Thirty-eight S. aureus were isolated. The results showed that six MRSA were detected with particularly interesting mecC-carrying MRSA isolates which had not yet been found in Portugal. A low frequency of antibiotic resistance and virulence genes was observed among the isolates. Nevertheless, a high diversity of clonal lineages was detected among S. aureus some of which are associated with livestock.The frequent carriage of Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), by wild animals along with its zoonotic potential poses a public health problem. Furthermore, the repeated detection of the mecA gene homologue, mecC, in wildlife raises the question whether these animals may be a reservoir for mecC-MRSA. Thus, we aimed to isolate S. aureus and MRSA from wild rodents living in port areas and to characterize their antimicrobial resistance and genetic lineages. Mouth and rectal swab samples were recovered from 204 wild rodents. The samples were incubated in BHI broth with 6.5% of NaCl and after 24 h at 37 °C the inoculum was seeded onto Baird-Parker agar, Mannitol Salt agar and ORSAB (supplemented with 2 mg/L of oxacillin) plates. Species identification was confirmed by MALDI-TOF MS. The antimicrobial susceptibility testing was performed by the Kirby–Bauer disc diffusion method against 14 antibiotics. The presence of virulence and resistance genes was performed by PCR. The immune evasion cluster (IEC) system was investigated in all S. aureus. All isolates were characterized by MLST, spa- and agr typing. From 204 samples, 38 S. aureus were isolated of which six MRSA were detected. Among the six MRSA isolates, three harbored the mecC gene and the other three, the mecA gene. All mecC-MRSA isolates were ascribed to sequence type (ST) 1945 (which belongs to CC130) and spa-type t1535 whereas the mecA isolates belonged to ST22 and ST36 and spa-types t747 and t018. Twenty-five S. aureus were susceptible to all antibiotics tested. S. aureus isolates were ascribed to 11 MLST and 12 spa-types. S. aureus presents a great diversity of genetic lineages in wild rodents. This is the first report of mecC-MRSA in Portugal.

Clonal diversity and genomic characterization of Panton-valentine Leukocidin (PVL)-positive Staphylococcus aureus in Tehran, Iran

BackgroundSome Staphylococcus aureus strains produce Panton-Valentine leukocidin (PVL), a bi-component pore-forming toxin, which causes leukocyte lysis and tissue necrosis. Currently, there is very limited information on the molecular epidemiology of PVL-encoding S. aureus strains in Iran. This study aimed to determine the molecular epidemiology and genetic background of PVL-positive S. aureus clinical strains isolated from Iranian patients.MethodsA total of 28 PVL-positive S. aureus strains were detected from 600 S. aureus isolates between February 2015 and March 2018 from different hospitals in Tehran, Iran. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Molecular genotyping was performed using SCCmec and accessory gene regulator (agr) typing, PVL haplotyping, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE).ResultsThe highest antibiotic resistance rate was found to be against erythromycin (57.1%), followed by ciprofloxacin (42.8%) and clindamycin (35.7%). Moreover, 19 (67.9%) out of 28 S. aureus isolates were identified as MRSA, including CA-MRSA (14/19, 73.7%) and HA-MRSA (5/19, 26.3%). SCCmec type IVa was detected as the predominant type (10/19, 52.6%), followed by type III (5/19, 26.3%) and type V (4/19, 21.1%). The agr type I was identified as the most common type (14/28, 50%), and H and R haplotype groups were observed at frequencies of 67.9 and 32.1%, respectively. Among H variants, the predominant variant was H2 (78/9%). The isolates encompassed 21 different sequence types (STs), including 16 new STs (ST5147 to ST5162). Based on eBURST analysis, the isolates were clustered into five CCs, including CC30, CC22, CC1, CC8, and CC5 (ST5160), and nine singletons. PFGE typing showed that 24 isolates were clustered into A (4 pulsotypes), B (9 pulsotypes), and C (11 pulsotypes) clusters.ConclusionsA high prevalence of PVL-positive CA-MRSA strains was detected in Iran. The majority of PVL-positive isolates were of H (mostly H2) variant, while R variant was harbored by 100% of PVL-positive MRSA strains. Also, CC8, CC22, and CC30 were identified as the dominant clones among PVL-encoding S. aureus strains. This study promotes a better understanding of the molecular epidemiology and evolution of PVL-positive S. aureus strains in Iran.

Correlations of Host and Bacterial Characteristics with Clinical Parameters and Survival in Staphylococcus aureus Bacteremia.

Staphylococcus aureus bacteremia (SAB) is a frequent, severe condition that occurs in patients of all age groups and affects clinical departments of all medical fields. It is associated with a high mortality rate of 20–30%. In this study, we analyzed patient mortality associated with SAB at our tertiary care university hospital, assessed the clinical management in terms of administered antimicrobial therapy, and determined which factors have an impact on the clinical course and outcome of patients with this disease. We collected clinical data and blood culture isolates of 178 patients diagnosed with SAB between May 2013 and July 2015. For this study, bacteria were cultured and analyzed concerning their phenotype, hemolysis activity, biofilm formation, nuclease activity, prevalence of toxin genes, spa and agr type. Overall mortality was 24.2% and 30-day mortality was 14.6%. Inadequate initial therapy was administered to 26.2% of patients and was associated with decreased survival (p = 0.041). Other factors associated with poor survival were patient age (p = 0.003), agr type 4 (p ≤ 0.001) and pathological leukocyte counts (p = 0.029 if elevated and p = 0.003 if lowered). The type of infection focus, spa clonal complex and enterotoxin genes seg and sei had an impact on severity of inflammation. Our results indicate that mortality and burden of disease posed by SAB are high at our university hospital.

Biofilm formation, icaABCD genes and agr genotyping of Staphylococcus aureus from fish and ground beef

AbstractA total of 46 Staphylococcus aureus isolates from fish and ground beef were tested for the agr types, icaABCD genes, and biofilm formation at 12, 25 and 37 °C by the microtiter plate and the MTT assays. All isolates were positive for the icaABD genes, while 97.8% were positive for the icaC. All isolates produced biofilms at 37 and 25 °C, but 93.5% of them were also biofilm producers at 12 °C. There was no significant difference in biofilm formation between 25 and 37 °C using the crystal violet assay (P &gt; 0.05). However, statistically significant differences were detected between 12 and 25 °C as well as 12 and 37 °C (P &lt; 0.05). All isolates were significantly different in biofilm production by the MTT assay at all tested temperatures. Furthermore, a relationship between the presence of the icaABCD genes and biofilm formation was observed. The agr type I was the most prevalent (54.4%) among the isolates, followed by agr type II (41.3%) and agr type III (9.6%). In this study, the S. aureus isolates exhibited biofilm formation ability responsible for persistence of bacteria in foods, which may lead to food spoilage and human health problems.

Clinical and molecular characteristics of Staphylococcus aureus isolated from Chinese children: association among the agr groups and genotypes, virulence genes and disease types.

This study was aimed to investigate the clinical and molecular epidemiology of Staphylococcus aureus (S. aureus) isolated from Chinese children and determine the possible relationship among the accessory gene regulator (agr) groups and genotypes, as well as among the virulence genes and disease types. S. aureus strains were isolated from Beijing Children's Hospital between October 2017 and October 2019. The isolates and 19 virulence genes were characterized using multi-locus sequence typing, staphylococcal protein A (spa), staphylococcal cassette chromosome mec, and agr typing. A total of 191 non-repetitive S. aureus clinical isolates were divided into 33 sequence types (STs), 18 clonal complexes (CCs), and 59spatypes. ST59 (39.8%), t437 (37.7%), and agr I (84.8%) were the predominant types. CC59, CC25, CC22, CC951, CC8, and CC398 belonged to agr I. CC5 and CC15 were assigned to agr II, and CC30 was characterized as agr III. CC121 was classified under agr IV. The eta, etb, and bbp genes were more prevalent in agr IV (P < 0.001 for each), while tst was more prevalent in agr group III compared to the other groups (P < 0.001). Nearly all isolates that harbored lukS/F-PV belonged to agr I (P = 0.005). However, the correlation between disease types and agr groups was not significant (P > 0.05). An association among the agr groups and genotypes, as well as specific toxin genes, was observed among the S. aureus strains isolated from Chinese children. However, a statistical correlation was not found among the agr groups and disease types.

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.

Molecular characterization and genetic diversity of Staphylococcus aureus isolates of dairy production farms in Rio de Janeiro, Brazil

Staphylococcus aureus is an important pathogen involved in subclinical bovine mastitis, causing high economic losses for the dairy industry. The successful persistence of this pathogen in the host occurs due to a series of factors associated with colonization ability and the acquisition of virulence factors. This bacterial species carries genetic heterogeneity, and genetically diverse strains characterize the population. Analysis of the genetic variation is an important tool for epidemiological studies. For this study, S. aureus strains were randomly selected by molecular profiling. All strains were originated from the milk of cows of subclinical mastitis on farms in the State of Rio de Janeiro. Strains of S. aureus were profiled using virulence gene analysis profiles, agr and spa typing, Pulsed Field Gel Electrophoresis (PFGE), and Multilocus Sequence Typing (MLST). 47% (8/17) of the tested strains were positive strains for icaA gene; 82.3% (14/17) for icaD gene; 41% (7/17) for fbnA gene; 47% (8/17) positive for fbnB gene; 94% (16/17) for the hlA gene and 70.5% (12/17) for hlB gene. These virulence results generated 11 different profiles. Most strains (58.8% - 10/17) were classified as type-II by agr system. spa typing identified seven different spa types. PFGE analysis found extensive genetic heterogeneity and no clones were observed. MLST analysis generated five different types of ST/CC. Considering the results observed at the present study, a high genetic variety of S. aureus strains associated of the presence of different virulence factors justified the absence of clonal strains at the properties evaluated. Besides, that, the definition of S. aureus clonal strain as well dynamic population in not fully understood since there are a limited number of studies in S. aureus associated with bovine mastitis.

Short communication: Association between the accessory gene regulator (agr) group and the severity of bovine mastitis caused by Staphylococcus aureus

Staphylococcus aureus can elicit mild to more severe degrees of mastitis in cattle, depending on the response of the host's immune system and the virulence factors of the specific isolate. Several virulence factors are controlled by a global regulatory system, designated accessory gene regulator (agr). Thus, the objective was to examine associations between different capsular and agr types and the severity of bovine mastitis caused by S. aureus. All isolates were obtained from bovine subclinical (n = 50), mild clinical (n = 73), and moderate clinical mastitis cases (n = 28). Isolates containing the agrI gene and lacking the agr locus (agr-) were more prevalent among subclinical than clinical mastitis cases, whereas isolates containing the agrII and agrIII genes were more prevalent among clinical mastitis cases. The capsular types 5 (cap5) and 8 (cap8) were found in 42 and 44%, respectively, of the isolates obtained from subclinical cases and in 38.6 and 58.4%, respectively, of those isolated from clinical mastitis cases. Capsular type was not associated with type of mastitis (subclinical, mild clinical, or moderate clinical). We found a strong association between agr type and type of mastitis, suggesting that knowledge of S. aureus genetic profiles could be an additional tool to control this disease.

Molecular typing, biofilm formation, and analysis of adhesion factors in Staphylococcus aureus strains isolated from urinary tract infections

Staphylococcus aureus is considered one of the principal bacteria challenges in public health today and is found particularly in patients with urinary tract infections. The data is still insufficient regarding biofilm-forming strains isolated from UTIs in Iran. In the present survey, 110 S. aureus strains were detected by antimicrobial susceptibility testing and biofilm formability. S. aureus isolates were also further investigated by carriage of biofilm and adhesion genes, and accessory gene regulator (agr) specificity groups using PCR and multiplex PCR. Out of 110 isolates tested, MRSA and MSSA accounted for 56.4% and 43.6%, respectively. Seventy-seven (70%) isolates were established as biofilm makers with complete dominant clfA, clfB genes and a markedly high prevalence of fnbA (80%), followed by fnbB (77.3%), sdrC (68.2%), icaA (62.7%), icaD (58.2%) eno (58.2%), sdrD (53.6%), ebps (38.2%), cna (25.5%), fib (20%), sdrE (17.3%), and bap (3.6%). A total of 4 agr types; type III (54.5%), II (20.9%), I (18.2%) and IV (6.4%) were detected in this study. The strong biofilm producers were assigned to particular agr type I (3.9%, 3/77), IV (1.3%, 1/77), and III (57.1%, 44/77), moderate biofilm producers to agr types I, II, and III, and weak biofilm producers to agr type II. Our findings illustrate corrections between biofilm formation and agr type III. We detected a high rate of MDR and biofilm formation in MRSA and MSSA strains isolated from UTI in varying degrees, which confirms the act of formation of biofilm in promoting antibiotic resistance.

Molecular characterization of invasive Staphylococcus aureus strains isolated from patients with diabetes in Iran: USA300 emerges as the major type.

There have been few studies focused on the molecular characterization of invasive Staphylococcus aureus strains in patients with diabetes in Iran. In the present study, 20 invasive S. aureus strains recovered from the patients with diabetes characterized by the virulence and resistance analysis, biofilm formation, staphylocoagulase (SC) typing, S. aureus protein A locus (spa) typing staphylococcal cassette chromosome mec (SCCmec) typing, and multilocus sequence typing (MLST). Virulence gene detection indicated a high prevalence of strains encoding the pvl genes (50%), a low prevalence of the tst and seg gene (each of them was 5%) and a markedly high prevalence of fnbB (95%), fnbA (85%), icaD (75%), icaA (65%). A total of 3 coagulase types (III, 85%; II, 10%; V, 5%), 2 agr types (I, 90%; II 10%) and 2 SCCmec types (IV, 65%; III, 35%) and four different clones namely ST8-MRSA-IV/t008 (50%) (USA300), ST239-MRSA-III/t030 (35%), ST5-MRSA-IV/t002 (10%), and ST45-MRSA-IV/t038 (5%) were detected in this study. Eighty-five percent of the isolates were biofilm producers. All the 4 high-level mupirocin resistance (HLMUPR) strains belonged to CC/ST8-MRSA-IV/t008 clone and carried mupA gene. Fusidic acid-resistant isolate belonged to ST239-SCCmec III/t030 clone. One vancomycin-intermediate resistance isolates was detected in our study, which belonged to ST5-MRSA-IVt002. Circulating clone in MRSA strains (USA300) isolated from the patients with diabetes highlighting the possibility of transmission of these microorganisms' clones between hospital, community, and environments. However, further studies require providing critical insights into the importance of continued controlling and treatment of S. aureus infections in patients with diabetes.

First report of methicillin-resistant Staphylococcus aureus carrying the mecC gene in human samples from Iran: prevalence and molecular characteristics

There is a lack of information concerning mecC clinical methicillin-resistant Staphylococcus aureus (MRSA) strains throughout the world. In the present survey, 345 MRSA strains were characterized by antimicrobial susceptibility testing and staphylococcal cassette chromosome mec element (SCCmec) typing. mecC-positive MRSA isolates were characterized by study of biofilm formability, adhesion and virulence analysis, multilocus sequence typing, accessory gene regulator (agr) typing, S. aureus protein A locus (spa) typing and staphylocoagulase typing. The present study found ten SCCmec types, with the majority being SCCmec type III (38.3%). The presence of mecC was confirmed in three isolates from skin wounds (two isolates) and burn wounds (one isolate). All the mecC-positive isolates carried SCCmec XI and belonged to coa type III. Molecular typing showed that these isolates belonged to clonal complex/ST130-spa type t843-agr type III (two isolates) and clonal complex/ST599-spa type 5930-agr type I. The presence of SCCmec type IV confirms the hypothesis of extensive infiltration from the community to the hospital. Detection of MRSA isolates harbouring the mecC gene highlights the need to perform routine detection methods and molecular investigations in order to identify these emerging strains and limit their transfer in hospitals and communities.

Prevalence and molecular epidemiology of ceftaroline non-susceptible methicillin-resistant Staphylococcus aureus isolates, first clinical report from Iran.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major pathogens in Iran with a high prevalence and a high level of antibiotic resistance. Ceftaroline is a fifth generation cephalosporin binding and inhibiting penicillin binding protein (PBP2a). In the present study, 228 clinical MRSA isolates were collected from four cities of Iran and their susceptibility to ceftaroline was evaluated by E-test and the disk diffusion method. Our results showed a high susceptibility rate (97.3%) to ceftaroline in MRSA strains from Iran. Six isolates were found to be ceftaroline non-susceptible (CPT-NS) with Minimum inhibitory concentration (MIC) ≥2 µg/mL. All CPT-NS isolates were isolated from blood and tracheal aspirate and belonged to SCCmec type III as well as agr type I and were all susceptible to vancomycin. Out of six isolates, three, two and one belonged to spa type t030, t4864, and t969, respectively. Vancomycin, quinupristin/dalfopristin, linezolid, chloramphenicol, and tigecycline were the most active agents against CPT-NS isolates. Due to the broad-spectrum activity and low toxicity of ceftaroline as well as the increased rate of vancomycin resistance among MRSA strains in recent years, ceftaroline can be considered as a novel approach to treat MRSA-induced infections.