Methicillin-sensitive S. Aureus Isolates Research Articles

Microbiological analysis of skin and soft tissue infections in children presenting to a paediatric tertiary care centre in Sri Lanka

Introduction: Skin and soft tissue infections (SSTIs) are a common cause of hospitalisation in children. They are diverse in spectrum, ranging from impetigo and abscesses to life threatening necrotising infections. The main pathogens associated with SSTIs are Staphylococccus aureus and Streptococcus pyogenes. Emergence of community acquired methicillin resistant S. aureus (CA-MRSA) is of major concern when antibiotics are required, and it is important to know the prevalence of these pathogens in different geographic locations. This study was done with the aim of identifying and characterising the common bacterial pathogens implicated in SSTIs in children presenting to Sirimavo Bandaranaike Children’s Hospital (SBSCH), Peradeniya, Sri Lanka, which is one of the two paediatric referral hospitals in Sri Lanka.Method: A retrospective, descriptive study analysed data gathered from request forms accompanying pus and tissue sent for culture from patients with SSTIs to the SBSCH microbiology laboratory during a period of 13 months from January 2021 to January 2022.Results: Two hundred and ninety samples were received from suspected SSTIs of which 197 (67.9%) were positive for bacterial growth, with S. aureus present in 163 (83%). Methicillin resistant S. aureus (MRSA) was isolated from 113 (57%) and methicillin sensitive S. aureus (MSSA) from 50 (25%). The frequency of coliforms, Pseudomonas spp., and Group A streptococcus were 31 (16%), 3 (1.5%) and 4 (2%) respectively. Sensitivity of MRSA isolates to cotrimoxazole, clindamycin, ciprofloxacin and erythromycin were 97%, 86%,72% and 29% respectively. Among MSSA isolates, 94% were sensitive to cotrimoxazole, while sensitivity to clindamycin, ciprofloxacin and erythromycin were 74%, 67% and 40% respectively.Conclusion: MRSA was the most frequent pathogen associated with purulent SSTIs in children, followed by MSSA. One fourth of MSSA were resistant to ciprofloxacin and clindamycin. Ciprofloxacin and clindamycin resistance in MRSA was 15% and 28% respectively.

Molecular Characteristics, Antimicrobial Susceptibility, Biofilm-Forming Ability of Clinically Invasive Staphylococcus aureus Isolates.

This study aimed to investigate the molecular characteristics, antimicrobial resistance, and biofilm-forming ability of Staphylococcus aureus isolates from invasive infections. A total of 92 non-repetitive S. aureus isolates from invasive infections were analyzed by Multi-locus Sequence Typing (MLST), spa typing, and chromosomal cassette mec (SCCmec) typing. Antibiotic susceptibility testing was performed using the disk diffusion and agar dilution methods. Biofilm-forming ability was assessed using crystal violet assay. The presence and expression of biofilm-associated genes were examined using PCR and RT-qPCR. Among the 55 Methicillin-resistant S. aureus (MRSA) and 41 Methicillin-sensitive S. aureus (MSSA) isolates, ST59 (43.6%) predominated in MRSA, while ST7 (39.0%) was most common in MSSA. As expected, MRSA exhibited higher antibiotic resistance rates compared to MSSA isolates. Biofilm formation assays revealed that the majority of isolates (88.5%) produced biofilms, with 26.0% classified as strong producers (OD570 ≥ 1.0) and 62.5% as weak producers (0.2 ≤ OD570<1.0). MSSA exhibited a higher biofilm-forming ability than MRSA (P < 0.01), with variations across clones. Notably, ST7 isolates displayed greater biofilm-forming ability than other sequence types (ST59, ST5, and ST239). RT-qPCR results revealed that ST7 isolates exhibited higher expression levels of icaA compared to other sequence types. This study revealed significant molecular heterogeneity among invasive S. aureus isolates, with ST59 and ST7 as dominant clones. The strong biofilm-forming capacity of ST7 merits concern given its rising prevalence regionally. Continuous surveillance of emerging successful lineages is critical to help guide infection control strategies against invasive S. aureus infections.

Structural and Antibacterial Characterization of a New Benzamide FtsZ Inhibitor with Superior Bactericidal Activity and In Vivo Efficacy Against Multidrug-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a multidrug-resistant (MDR) bacterial pathogen of acute clinical significance. Resistance to current standard-of-care antibiotics, such as vancomycin and linezolid, among nosocomial and community-acquired MRSA clinical isolates is on the rise. This threat to global public health highlights the need to develop new antibiotics for the treatment of MRSA infections. Here, we describe a new benzamide FtsZ inhibitor (TXH9179) with superior antistaphylococcal activity relative to earlier-generation benzamides like PC190723 and TXA707. TXH9179 was found to be 4-fold more potent than TXA707 against a library of 55 methicillin-sensitive S. aureus (MSSA) and MRSA clinical isolates, including MRSA isolates resistant to vancomycin and linezolid. TXH9179 was also associated with a lower frequency of resistance relative to TXA707 in all but one of the MSSA and MRSA isolates examined, with the observed resistance being due to mutations in the ftsZ gene. TXH9179 induced changes in MRSA cell morphology, cell division, and FtsZ localization are fully consistent with its actions as a FtsZ inhibitor. Crystallographic studies demonstrate the direct interaction of TXH9179 with S. aureus FtsZ (SaFtsZ), while delineating the key molecular contacts that drive complex formation. TXH9179 was not associated with any mammalian cytotoxicity, even at a concentration 10-fold greater than that producing antistaphylococcal activity. In serum, the carboxamide prodrug of TXH9179 (TXH1033) is rapidly hydrolyzed to TXH9179 by serum acetylcholinesterases. Significantly, both intravenously and orally administered TXH1033 exhibited enhanced in vivo efficacy relative to the carboxamide prodrug of TXA707 (TXA709) in treating a mouse model of systemic (peritonitis) MRSA infection. Viewed as a whole, our results highlight TXH9179 as a promising new benzamide FtsZ inhibitor worthy of further development.

Whole-genome analysis of methicillin-resistant and methicillin-sensitive Staphylococcus aureus in dry-fermented salami

Dry-fermented salami (DFS) is a popular and frequently consumed ready-to-eat food product. The safety of these products is typically ensured by the addition of well-defined starter cultures and subsequent fermentation processes. This study aimed to evaluate the prevalence of Staphylococcus aureus in DFS, investigate antimicrobial resistance and selected virulence genes of the isolates obtained, and estimate the possible source of S. aureus contamination in the final products based on their detailed characteristics. A total of five multidrug-resistant methicillin-resistant (MRSA) and sixteen methicillin-sensitive S. aureus (MSSA) isolates were characterised by whole genome sequencing. A total of eight different ST types and nine spa types were identified, with ST398-t011 predominating in MRSA and ST7-t091 in MSSA isolates. Methicillin-resistant ST398 isolates were classified as livestock-associated MRSA (LA-MRSA), and we hypothesized that the source of product contamination was raw pork. Various genes encoding staphylococcal enterotoxins, virulence factors, and factors mediating host invasion and immune evasion were identified exclusively in MSSA isolates, their origin may therefore be seen in human carriers. The results suggest the importance of high standards of Good Hygienic Practices and Good Manufacturing Practices in food processing.

The prevalence, genetic diversity and antibiotic resistance of Staphylococcus aureus associated with subclinical bovine mastitis in Balikesir, Turkey

Subclinical mastitis caused by Staphylococcus aureus is very common in dairy cows and creates serious problems on dairy farms. In this study, we investigated the prevalence, genetic diversity and antimicrobial resistance of S. aureus from subclinical bovine mastitis on 12 dairy herds in Balikesir province of Turkey, by SCCmec and spa typing. Ninety-five isolates of S. aureus were isolated from 725 subclinical mastitic milk samples that exceeded the somatic cell count (SCC) limit of 4 x 105 cell/mL. The frequency of MRSA (methicillin-resistant Staphylococcus aureus) and MSSA (methicillin-sensitive S. aureus) was 6.3% (6 isolates) and 93.68% (89 isolates) respectively. SCCmec types of MRSA isolates were community-associated CA-MRSA type IVb (4 isolates) and type IVd (two isolates), while the spa types were T 005 and T 5163 (three isolates from each). The resistance rate of MRSA isolates was 100% for oxacillin and cefoxitin, 83% for penicillin, ampicillin, clindamycin, erythromycin and 66% for gentamicin and trimethoprim/sulfamethoxazole. However, compared to MRSA, the resistance of MSSA isolates was relatively lower. This study supported the scientific data on the occurrence of MRSA and MSSA in subclinical mastitis, and highlighted the need for preventive measures to eliminate or decrease S. aureus contamination of milk in dairy herds.

Staphylococcus aureus in Non-Cystic Fibrosis Bronchiectasis: Prevalence and Genomic Basis of High Inoculum β-Lactam Resistance.

Rationale: The pathobiology of Staphylococcus aureus in non-cystic fibrosis bronchiectasis (nCFB) is poorly defined. When present at high density or "inoculum," some methicillin-sensitive S. aureus (MSSA) can inefficiently degrade antistaphylococcal β-lactam antibiotics via BlaZ penicillinases (termed the "inoculum effect" [IE]). Given the high burden of organisms in bronchiectatic airways, this is particularly relevant. Objectives: Drawing from a prospectively collected biobank, we sought to understand the prevalence, natural history, potential for transmission, and antibiotic resistance profiles among nCFB-derived MSSA isolates. Methods: All individuals attending a regional consultancy nCFB clinic with sputum collected between 1981 and 2017 were considered, and those with one or more S. aureus-positive cultures composed the cohort. Each individual's most recent biobank isolate was subjected to whole-genome sequencing (including the blaZ gene), antibacterial susceptibility testing, and comparative β-lactam testing at standard (5 × 105 colony-forming unit [cfu]/ml) and high (5 × 107 cfu/ml) inocula to assess for the IE and pronounced IE. Results: Seventy-four (35.4%) of 209 individuals had one or more sputum samples with S. aureus (68 MSSA, 6 methicillin-resistant S. aureus). Those with S. aureus infection were more likely to be female. Among 60 of 74 MSSA isolates subjected to whole-genome sequencing, no evidence of transmission was identified, although specific multilocus sequence typing types were prevalent, including ST-1, ST-15, ST-30, and ST-45. Antibiotic resistance was uncommon, except for macrolides (∼20%). Among the 60 MSSA samples, the prevalence of IE and pronounced IE was observed to be drug specific: meropenem (0% and 0%, respectively), cefepime (3% and 5%, respectively), ceftazidime (8% and 0%, respectively), cloxacillin (12% and 0%, respectively), cefazolin (23% and 0%, respectively), and piperacillin-tazobactam (37% and 17%, respectively). The cefazolin IE was associated with blaZ type A (P < 0.01) and ST-30 (P < 0.01), whereas the piperacillin-tazobactam IE was associated with type C blaZ (P < 0.001) and ST-15 (P < 0.05). Conclusions:S. aureus infection was common, although no evidence of transmission was apparent in our nCFB cohort. Although routine susceptibility testing did not identify significant resistance, inoculum-related resistance was found to be relevant for commonly used nCFB antibiotics, including cefazolin and piperacillin-tazobactam. Given previous associations between IEs and negative patient outcomes, further work is warranted to understand how this phenotype impacts nCFB disease progression.

Correlation between mazEF Toxin-Antitoxin System Expression and Methicillin Susceptibility in Staphylococcus aureus and Its Relation to Biofilm-Formation.

Methicillin resistance in Staphylococcus aureus has become prevalent globally. Moreover, biofilm-formation makes it more difficult to eradicate bacteria by antibiotics. The mazEF toxin-antitoxin system encodes for mazF, which acts as an endoribonuclease that cleaves cellular mRNAs at specific sequence motifs (ACA), and mazE, which opposes the mazF action. Our goal was to detect mazEF expression in methicillin-resistant S. aureus (MRSA) isolates compared with methicillin-sensitive S. aureus (MSSA) isolates and determine its relation to methicillin susceptibility as well as biofilm-formation. According to their susceptibility to cefoxitin disks, 100 S. aureus isolates obtained from patients admitted to Cairo University Hospitals were categorized into 50 MSSA and 50 MRSA according to their susceptibility to cefoxitin disks (30 µg). Antimicrobial susceptibility and biofilm-formation were investigated using the disk diffusion method and tissue culture plate method, respectively. Finally, using real-time PCR, mazEF expression was estimated and correlated to methicillin susceptibility and biofilm formation. Both MRSA and MSSA isolates showed the best sensitivity results with linezolid and gentamicin, where about 88% of MRSA isolates and 96% of MSSA isolates were sensitive to linezolid while 76% of MRSA isolates and 84% of MSSA isolates were sensitive to gentamicin. MRSA isolates were significantly more able to form biofilm than MSSA isolates (p-value = 0.037). The mazEF expression was significantly correlated to methicillin resistance in S. aureus (p-value < 0.001), but not to biofilm-formation.

The Prevalence of Inducible Clindamycin Resistance Staphylococcus aureus among Various Clinical Specimens in Khartoum state Sudan

Background : S. aureus is frequently associated with skin infections, pneumonia, surgery wounds, bacteraemia, osteomyelitis and endocarditis, being considered one of the most important pathogens of the human being, both at the community level and at nosocomial infections, and may become serious if caused by antimicrobial resistant strains, especially methicillin-resistant S. aureus (MRSA) strains, which are resistant to most of the antimicrobial agents, methicillin-sensitive S.aureus (MSSA) and isolates with reduced susceptibility and resistance to vancomycin, which is the last drug for the treatment of MRSA infections. So, this study aimed to detect the existence of inducible resistance of S. aureus to Clindamycin in Khartoum-Sudan among patients attended to Suba University Hospital. Methods : The study was performed as cross-sectional one, 53 clinical isolates of S. aureus obtained from (34 females and 19 males) with different clinical condition among patients attended to Suba University Hospital in Khartoum-Sudan from April to August 2017. To detect inducible clindamycin resistant by using D test. In addition to that MRSA / MSSA all the isolates screened for methicillin resistant by using 1 µg oxacillin then examined for inducible clindamycin resistant by D test. In addition to that examine for antimicrobial susceptibility profile which include vancomycin, gentamycin, tetracycline and co-trimoxazole. The data were analyzed using Statistical Package for Social Science, version 22, P. value &lt;0.05 was considered statistically significant Results : out of 53 isolates, 36 S. aureus isolated resistant to Clindamycin, 26 (72.2%) were MRSA and 10 (27.8%) were MSSA by means of D test, while 17 (32.1%) of isolates were sensitive 9 (53%) MRSA and 8 (47 %) MSSA. Comparing Induced clindamycin resistance showed equally distribution among MSSA and MRSA isolates, giving no significant difference as P- value 0.167. Conclusion : This study showed that D.test to detect inducible clundamycin resistance in staphylococcus aureus.

Molecular Characterization of Staphylococcus aureus Obtained from Blood Cultures of Paediatric Patients Treated in a Tertiary Care Hospital in Mexico.

PurposeStaphylococcus aureus is one of the main causative agents of hospital-acquired (HA) infections. In Mexico, information about the characteristics of clinical S. aureus isolates is limited. Our aim was to characterize S. aureus strains obtained from blood cultures of paediatric patients treated in a tertiary care hospital.Materials and MethodsWe analysed 249 S. aureus isolates over the period from 2006 to 2019, and their resistance profiles were determined. The isolates were classified into methicillin-resistant S. aureus (MRSA) or methicillin-sensitive S. aureus (MSSA). Staphylococcal cassettes chromosome mec (SCCmec) were detected. Virulence genes (cna, clfA, clfB, eta, etb, fnbA, fnbB, hla, pvl, sec, and tsst) were amplified, and their clonal relationships were established by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and clonal complex (CC) typing. We reviewed one hundred medical files to collect clinical information.ResultsThirty-eight percent of the isolates were MRSA and showed an expanded profile of resistance to other non-beta-lactam antibiotics, while MSSA strains presented a reduced resistance profile. SCCmec-II was the most frequent element (86.3%). Eight virulence factors were detected in MSSA and six in MRSA. The pvl gene was detected in four MRSA-SCCmec-IV isolates (P≤0.0001). MRSA isolates were distributed among 14 clones and were classified into 15 sequence types (ST); the most frequent was ST1011 (17%). The most common CC in MRSA was CC5 (69%, P≤0.0001), and in MSSA, it was CC30 (30%, P≤0.0001). Eighty-seven percent of MRSA isolates were HA-MRSA, and 13% were community-acquired MRSA (CA-MRSA). Of 21 HA-MRSA isolates, 17 had SCCmec-II, while two CA-MRSA isolates had SCCmec-IV. Of MSSA isolates, 77% were derived from HA infections and 23% from CA infections.ConclusionMSSA isolates had more virulence factors. MRSA isolates were resistant to more non-beta-lactam antibiotics, and those with SCCmec-IV expressed a greater variety of virulence factors. Most S. aureus isolates belonged to CC5.

Is COVID-19 really sparing children?

Background Staphylococcus aureus is an important nosocomial pathogen worldwide, with two major Classes Methicillin resistant S. aureus (MRSA) and Methicillin sensitive S. aureus (MSSA). Aim To compare the distribution frequency and antimicrobial sensitivity of MRSA and MSSA S. aureus isolates in different clinical specimen from hospitalized Iraqi patients. Materials and Methods S. aureus isolates from clinical specimens were investigated in 203 hospitalizes patients with wide range of ages during the period from February to May 2017. API and Vitek were used for identification and a panel of antibiotics was used to define the antimicrobial sensitivity of the isolates. Results The highest S. aureus isolates were from burn swab (35%), followed by urine specimen and blood samples with (30 and 26% respectively). MSSA isolates represents (57.5%) of the total and the rest was MRSA isolates (42.5%). MRSA isolates was higher in burns and wound specimens (485 and 13% respectively) whereas the MSSA isolates were higher in blood, urine and ear specimens (29%, 38% and 3.5% respectively). MRSA were multidrug resistance to 7 antibiotics in comparison to MSSA (only two antibiotics). Conclusion MSSA isolate are more common than MRSA in clinical specimens with variable proportions in different clinical specimens. Multidrug resistance was more evident among the MRSA than MSSA.

Comparative study for the efficacy of Sofinox cream against Staphylococcus aureus with Fucidin cream, T-Bact ointment and screening for resistant mutants.

Objective: To determine the Minimum Inhibition Concentration of Sofinox cream, Fucidin cream, T Bact ointment against Staphylococcus aureus and to screen for the development of resistant mutants and Whole Genome Sequencing(WGS) of the strains used and the strains showing raised MIC following serial passages. Material and Methods: An in vitro study was conducted to determine the Minimum Inhibition Concentration of Sofinox cream, Fucidin cream, T Bact ointment against seven strains of Staphylococcus aureus (ATCC 25923, ATCC 43300, ATCC 700699, 2 clinical isolates of methicillin-sensitive S. aureus (MSSA) and 2 methicillin-resistant S. aureus (MRSA). We also screened the same strains for the development of resistant mutants of Staphylococcus aureus by sub MIC exposure to Sofinox cream, Fucidin cream and T Bact ointment up to 70 serial passage and their WGS. Results: The MIC values of the three topical antibiotics ranged between 16 µg to 64 µg for all the strains of Staphylococcal strains tested. The MIC value did not change for three strains of Staphylococcus aureus and a one-fold rise in MIC value occurred for four strains of the Staphylococcus aureus strains for Sofinox cream after 70 passages at sub MIC concentration. WGS analysis of the strains with Sofinox cream treatment, it was noted that 150 number significant genetic variations to MSSA (ATCC 25923) and 84 number for MRSA (ATCC 700699) and no significant changes with other strains. For two of the strains the MIC value remained the same, three strains showed the one-fold rise and two strains showed a four-fold rise in MIC value for Fucidin cream after 70 passages at sub MIC concentration. Through the WGS analysis of the strains, it was noted that 185 significant genetic variations to one of the clinical isolates of MSSA followed by 102 to MRSA standard strain ATCC700699, the strains showing a one-fold rise in MIC showed 95, 85 and 76 number of genetic variations respectively. All the strains of Staphylococcus aureus showed a rise in MIC for T Bact which ranged from one to four-fold rise. WGS analysis of the strains with T Bact ointment treatment, it was noted the variations ranging from 85 to 158 with a one-fold rise to a four-fold rise in MIC value. Conclusions: In the present study it was observed that Sofinox cream had low resistance potential in vitro compared to Fucidin cream whereas T bact ointment had more resistance potential when exposed to sub MIC concentration of the cream/ointment. Even the WGS analysis showed variations correlating with MIC values. Hence there is a less likely chance of development of resistant mutants with the topical use of Sofinox cream even up to 8 weeks.

Characterisation of antibiotic resistance, virulence, clonality and mortality in MRSA and MSSA bloodstream infections at a tertiary-level hospital in Hungary: a 6-year retrospective study

BackgroundStaphylococcus aureus bloodstream infections (BSI) cause significant morbidity and mortality due to the frequent antibiotic resistance, toxin and adhesin production of the bacterium. These characteristics differ significantly in methicillin resistant (MRSA) and methicillin sensitive S. aureus (MSSA) and also among isolates of different MRSA clones, contributing to the outcome of S. aureus bacteraemia.MethodsIn this study, all MRSA BSI isolates from Semmelweis University, Budapest, Hungary, isolated between 2011–2016 and the same number of matched MSSA (overall 306 isolates) were characterised in terms of antibiotic susceptibility, virulence genes, clonality and their association with all-cause 30-day mortality. Effect of patient related variables, such as age, gender and comorbidities were also investigated.ResultsST22-MRSA-IV and ST5-MRSA-II were the most prevalent clones in our study. SCCmec I isolates showed the highest resistance rates and SCCmec II carried most virulence genes. Infections caused by SCCmec IV isolates were associated with the highest mortality rate (42.2%), despite the similar comorbidity rates of the different patient groups. All-cause 30-day mortality was 39.9% in the MRSA and 30.7% in the MSSA group. Increased teicoplanin MIC was associated with high mortality rate. Resistance to ciprofloxacin, erythromycin and clindamycin was common in MRSA, whereas MSSA isolates were more sensitive to all antibiotics with the exception of doxycycline. All MRSA isolates were sensitive to glycopeptides and linezolid; resistance to rifampicin and sulfamethoxazole-trimethoprim was low. MRSA isolates carried more adhesion genes, superantigens were more frequent in MSSA. Panton-Valentine leukocidin was found in 2.3% of the isolates.ConclusionThis study provides insight into the clonal composition and associated mortality of BSI S. aureus isolates in Hungary. The results suggest that the outcome of the infection is determined by the antibiotic resistance, genotype of the bacterium, and patient-related factors; rather than the virulence factors carried by the bacteria.

Prevalence, Genetic Diversity, and Temporary Shifts of Inducible Clindamycin Resistance Staphylococcus aureus Clones in Tehran, Iran: A Molecular-Epidemiological Analysis From 2013 to 2018.

The prevalence of Staphylococcus aureus as an aggressive pathogen resistant to multiple antibiotics causing nosocomial and community-acquired infections is increasing with limited therapeutic options. Macrolide-lincosamide streptogramin B (MLSB) family of antibiotics represents an important alternative therapy for staphylococcal infections. This study was conducted over a period of five years from August 2013 to July 2018 to investigate the prevalence and molecular epidemiology in Iran of inducible resistance in S. aureus. In the current study, 126 inducible methicillin-resistant S. aureus (MRSA) (n = 106) and methicillin-sensitive S. aureus (MSSA) (n = 20) isolates were characterized by in vitro susceptibility analysis, resistance and virulence encoding gene distribution, phenotypic and genotypic analysis of biofilm formation, prophage typing, S. aureus protein A locus (spa) typing, staphylocoagulase (SC) typing, staphylococcal cassette chromosome mec (SCCmec) typing, and multilocus sequence typing. Of the 126 isolates, 76 (60.3%) were classified as hospital onset, and 50 (39.7%) were classified as community onset (CO). Biofilm formation was observed in 97 strains (77%). A total of 14 sequence types (STs), 26 spa types, 7 coagulase types, 9 prophage types, 3 agr types (no agr IV), and 9 clonal complexes (CCs) were identified in this study. The prevalence of the inducible MLSB (iMLSB) S. aureus increased from 7.5% (25/335) to 21.7% (38/175) during the study period. The iMLSB MRSA isolates were distributed in nine CCs, whereas the MSSA isolates were less diverse, which mainly belonged to CC22 (7.95%) and CC30 (7.95%). High-level mupirocin-resistant strains belonged to ST85-SCCmec IV/t008 (n = 4), ST5-SCCmec IV/t002 (n = 4), ST239-SCCmec III/t631 (n = 2), and ST8-SCCmec IV/t064 (n = 2) clones, whereas low-level mupirocin-resistant strains belonged to ST15-SCCmec IV/t084 (n = 5), ST239-SCCmec III/t860 (n = 3), and ST22-SCCmec IV/t790 (n = 3) clones. All the fusidic acid–resistant iMLSB isolates were MRSA and belonged to ST15-SCCmec IV/t084 (n = 2), ST239-SCCmec III/t030 (n = 2), ST1-SCCmec V/t6811 (n = 1), ST80-SCCmec IV/t044 (n = 1), and ST59-SCCmec IV/t437 (n = 1). The CC22 that was predominant in 2013–2014 (36% of the isolates) had almost disappeared in 2017–2018, being replaced by the CC8, which represented 39.5% of the 2017–2018 isolates. This is the first description of temporal shifts of iMLSB S. aureus isolates in Iran that identifies predominant clones and treatment options for iMLSB S. aureus–related infections.

Spa gene-based molecular typing of nasal methicillin-susceptible staphylococcus aureus from patients and health-care workers in a dialysis center in southeast Iran

ABSTRACTStaphylococcus aureus is an important infectious agent in hemodialysis patients. This study was conducted to determine the frequency of S. aureus nasal carriage in hemodialysis patients (HD) and health-care workers (HCW) at the main dialysis center of Bam city, located in southeast of Iran. In this cross-sectional study, a total of 52 nasal swabs were obtained from health-care workers and hemodialysis patients to detect methicillin-sensitive S. aureus (MSSA) isolates. The resistance to different antibacterial agents was determined by disk diffusion method. Also, Panton-Valentine Leukocidin (PVL) – encoding gene as well as Staphylococcal protein A (spa) type were determined. The nasal carriage rate of S. aureus was found to be 24.4% and 18.8% in patients on hemodialysis and health-care workers, respectively. Among identified isolates, no methicillin-resistant S. aureus (MRSA) was found. Only two MSSA isolates (16.7%) were resistant to trimethoprim/sulfamethoxazole. One isolate (8.6%) was positive for pvl gene. Moreover, 8 spa types were found. According to BURP analysis, six out of the 12 S. aureus isolates (50%) belonged to the same clone, indicating a prevalence of a major clone among MSSA in carriage, including patients and HCW. Mupirocin is still the appropriate drug for reducing nasal colonization in our setting. Accumulation of isolates from patients and staff in one spa clonal complex is alarming for the necessity of more serious infection control in this center. Therefore, it is necessary to screen patients and health-care workers as a health priority, in order to prevent cross transmission.

Identification of ica-dependent biofilm production by Staphylococcus aureus clinical isolates and antibiofilm effects of ascorbic acid against biofilm production

AimsStaphylococcus aureus (S. aureus) is a life-threatening pathogen with high morbidity and mortality rates which causes nosocomial and community-acquired infections. Biofilm, considered to be a common virulence factor for pathogens,...

Prevalence of Methicillin-Resistant Staphylococcus aureus in Shrines.

Methicillin-resistant Staphylococcus aureus (MRSA) infection in human beings and animals is concerning; it stands out as one of the leading agents causing nosocomial and community infections. Also, marginally increasing drug resistance in MRSA has limited therapeutic options. This study focuses on estimating the prevalence of MRSA in shrines, a place where human and animal interaction is frequent, sharing antibiotic-resistant bacteria, antibiotic-resistant genes, and diseases. A total of 120 environmental swabs were collected from targeted areas during the study period, March 2018 to May 2018. Staphylococcus aureus was identified by growth on mannitol salt agar (MSA), and MRSA by growth on mannitol salt agar containing 4 μg Oxacillin, Gram staining, and conventional biochemical test. Isolates of S. aureus were characterized by antibiotic susceptibility testing using the disc diffusion method. MRSA and methicillin-sensitive S. aureus (MSSA) proportion were 19% and 81%, respectively; a high rate of MRSA was observed in isolates from Thapathali (28.6%). MSSA isolates showed a high rate of resistance to erythromycin (64.7%). MRSA isolates were resistant to gentamicin (50%), cotrimoxazole (25%), erythromycin (50%), and ciprofloxacin (25%). The isolates were susceptible to linezolid (100%), clindamycin (100%), ciprofloxacin (75%), erythromycin (50%), tetracycline (100%), and cotrimoxazole (75%). Intermediate resistance was also found in gentamicin (50%). Of the 11 MSSA isolates that were erythromycin resistant and clindamycin sensitive, 6 (54.5%) showed the inducible clindamycin resistance (ICR) pattern and 2 MRSA isolates that were erythromycin resistant and clindamycin sensitive showed ICR pattern. Fifteen MSSA isolates were β-lactamase positive, whereas only two MRSA isolates showed β-lactamase production. There exists a minimal research work on infectious diseases that are shared between primates and animals. This study suggests the pervasiveness of MRSA/MSSA in the shrines, which may be a primary place for pathogen exchange between humans and primates.

711. Exebacase (Lysin CF-301) Activity Against Staphylococcus aureus (S. aureus) Isolates From Bacteremic Patients Enrolled in a Phase 2 Study (CF-301-102)

BackgroundExebacase (CF-301) is a novel, recombinantly-produced, bacteriophage-derived lysin (cell wall hydrolase) which is the first lysin to report Phase 2 (Ph2) results which demonstrated 42.8% higher clinical responder rates with a single dose of exebacase used in addition to standard of care antibiotics (SOC) vs. SOC alone for the treatment of methicillin-resistant S. aureus (MRSA) bacteremia including endocarditis. We examined exebacase activity by broth microdilution (BMD) against baseline methicillin-sensitive S. aureus (MSSA) and MRSA isolates from each of the 116 participants in the recently complete exebacase”first in-patient’ Ph2 study (NCT03163446).MethodsPatients with complicated bacteremia or endocarditis caused by S. aureus were enrolled into Study CF-301-102 at study centers in the United States, EU, Latin America, Israel, and Russia from 2017 and 2018. Baseline isolates from blood cultures were collected prior to administration of exebacase. Exebacase MICs against 117 isolates of MSSA (n = 74) and MRSA (n = 43) were determined at a central laboratory using a modified BMD approved by the CLSI for exebacase AST.ResultsThe exebacase MICs of baseline patient isolates from the Ph2 study ranged from 0.125 – 2 µg/mL and the MIC50/90 values for all MSSA and MRSA isolates were 0.5/1 µg/mL. Exebacase MICs reported in a recent surveillance study were similar, with MIC50/90 values of 0.5/1 µg/mL. Of the 6 total subjects with EXE MICs of 2, 3 were clinical responders, 2 were indeterminate (not available for assessment), and 1 was a clinical nonresponder at Day 14.ConclusionExebacase was highly active against all baseline S. aureus isolates from blood cultures obtained from bacteremic patients enrolled in the Ph2 study. Based on data from previously presented exposure target attainment animal studies, PK/PD modeling and preliminary nonclinical breakpoint assessments, we expected that strains with MIC values of ≤2 µg/mL will have been susceptible to the Ph2 clinical exebacase dose determined based on target attainment studies under study in Ph2.Disclosures All authors: No reported disclosures.

Bacterial content and characterization of antibiotic resistant Staphylococcus aureus in Danish sushi products and association with food inspector rankings

This study examined the prevalence and phenotypic and genotypic antibiotic resistance patterns of Staphylococcus aureus in sushi from 20 Danish outlets. Microbial quality of sushi products and food inspector ranking of outlets were assessed and results for thirteen of the outlets were compared with findings from a previous study in 2012. Inspector rankings were similar in the two studies. The mesophilic aerobic counts were slightly lower (p = 0.0296) in 2017 than in 2012 with average values of the 13 shops of 5.2 log CFU/g and 5.7 log CFU/g, respectively. In both studies E. coli was only found in the products from outlets that did not have consistently superior rankings. On the other hand prevalence and average counts of Staphylococcus spp. were slightly higher in 2017 (p = 0.0286) but no methicillin-resistant S. aureus (MRSA) were observed in the present study. Methicillin-sensitive S. aureus (MSSA) were, however, isolated from 18.7% of sushi products with an average count below 2 log CFU/g. Based on spa and multilocus sequence typing (MLST), isolates belonged to clonal complex CC7 (t2016), CC20 (t7836), CC45 (t065, t127, t362), CC88 (t1998) and CC398 (t164, t331, t1451). The Panton-Valentine leukocidin (PVL)-encoding gene lukF was detected only in isolates of the t065 spa-type whereas the scn gene from the ΦSa3 prophage was detected in 76.5% of the isolates, supporting that the majority of isolates were of likely human origin. Thirty-six isolates (70.6%) were resistant to at least one of the antibiotic compounds tested. Antibiotic resistance genes that confer resistance to β-lactams (blaZ) and macrolides (ermC) were detected in 33.3% and 9.8% of isolates, respectively. The tet(K) gene that encode tetracycline resistance was only found in a t7836 strain. Overall, this study indicates that S. aureus in sushi products in Denmark do not represent a major food safety hazard due to, firstly, the low temperature and limited time of storage of product may prevent significant growth and production of toxic levels of enterotoxin of this species. Secondly, the S. aureus isolates obtained did not include MRSA variants and none of them encoded PVL that constitute one of the virulence factors in pathogenesis. Several MSSA isolates contained however genes encoding antibiotic resistance, which emphasize the potential role of foods as vehicles for transmission of such variants.

Accessory gene regulator (agr) dysfunction was unusual in Staphylococcus aureus isolated from Chinese children

BackgroundStaphylococcus aureus (S. aureus) with accessory gene regulator (agr) dysfunction occurs in health care settings. This study evaluated the prevalence and the molecular and drug resistance characteristics of S. aureus with dysfunctional agr in a pediatric population in Beijing, China.ResultsA total of 269 nonduplicate S. aureus clinical isolates were isolated from Beijing Children’s Hospital, including 211 methicillin-resistant S. aureus (MRSA) from September 2010–2017 and 58 methicillin-sensitive S. aureus (MSSA) from February 2016–2017. Only 8 MRSA and 2 MSSA isolates were identified as agr dysfunction, and the overall prevalence rate was 3.7%. For MRSA isolates, ST59-SCCmec IV and ST239-SCCmec III were the most common clones, and the prevalence rate of agr dysfunction in ST239-SCCmec III isolates (17.39%) was significantly higher than in ST59-SCCmec IV (1.69%) and other genotype strains (P = 0.006). Among the agr dysfunctional isolates, only one MRSA ST59 isolate and one MSSA ST22 isolate harbored pvl. No significant difference was detected between agr dysfunction and agr functional isolates regarding the biofilm formation ability (P = 0.4972); however, 9/10 agr dysfunctional isolates could effectuate strong biofilm formation and multidrug resistance. Among MRSA, the non-susceptibility rates to ciprofloxacin, gentamicin, and trimethoprim-sulfamethoxazole were significantly higher in agr dysfunctional isolates than in isolates with functional agr (P < 0.05). Two isolates belonging to ST239 had no mutations in agr locus, but a synonymous mutation was found in agrA in another ST239 isolate. The inactivating mutations were detected in other seven agr dysfunctional isolates. The variants were characterized by non-synonymous changes (n = 5) and frameshift mutations (insertions, n = 2), which mainly occurred in agrC and agrA.ConclusionsThe results showed that agr dysfunctional S. aureus was not common in Chinese children, and ST59-SCCmec IV was associated with lower prevalence of agr dysfunction as compared to ST239-SCCmec III isolates. The agr dysfunctional isolates were healthcare-associated, multidrug resistant and form strong biofilm, which suggested that agr dysfunction might offer potential advantages for S. aureus to survive in a medical environment.

Antibacterial effects of vancomycin in combination with methicillin against methicillin-resistant and methicillinsensitive Staphylococcus aureus

Background and aims: Staphylococcus aureus is considered as a frequent cause of infections. Therefore, antibacterial agents including β-lactam are normally used to treat these infections while the emergence of antibiotic resistance is one of the major clinical problems in this respect. Accordingly, combination antibacterial therapy is one way to alleviate this problem. As a result, the present study aimed to investigate the combined effects of vancomycin and methicillin on clinical isolates of methicillin-resistant S. aureus (MRSA) and methicillinsensitive S. aureus (MSSA). In addition, a series of follow-up studies were performed to evaluate the antibacterial activity. Methods: The current cross-sectional study was conducted on 40 hospitalized patients from various clinical samples such as pus/wound swabs, blood, urine and sputum during a 6-month period in 2017. To this end, the antibacterial activities of vancomycin and methicillin, alone and in combination were investigated against MRSA and MSSA. Eventually, the inhibitory effects of vancomycin and methicillin alone and in combination were studied on the growth profile of MRSA and MSSA, as well as the expression of mecA gene. Results: Based on the results, the significant synergistic and partial synergistic activity with fractional inhibitory concentration (FIC) and fractional bactericidal concentration (FBC) indexes ranged from 0.27-0.938 and 0.313-0.844, respectively, in the combination of vancomycin and methicillin in MRSA and MSSA isolates. Further, the obtained data indicated that the combination of vancomycin and methicillin had a synergistic effect against MRSA isolates (P&lt;0.01). The expression levels of the mecA gene were down-regulated by 5.25- fold in the combination of vancomycin and methicillin in MRSA isolate (P&lt;0.001). Conclusion: In general, these events may reflect the potential uses of vancomycin and methicillin combination against MRSA. However, a greater understanding of the underlying molecular mechanisms of vancomycin and methicillin in combination could contribute to the development of therapeutic strategies.