Methicillin-resistant Staphylococcus Aureus Strains Research Articles

Prominent Classes of Antibiotics and their Mechanism of Resistance against Methicillin-Resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) is a prominent pathogenic, antibiotic-resistant microorganism that contains a variety of virulent characteristics having the capacity to develop tolerance to several major classes of antibiotics. The ongoing creation of clones enhances this potential, transforming S. aureus into an “Anti-Infective.” MRSA has started to rise as a Hospital-Acquired MRSA, but due to evolution, new strains of MRSA have been discovered throughout the past several years. The new strains of MRSA as Community-Acquired MRSA, and Livestock-Associated MRSA are infecting the patients despite preexisting medical conditions, being as susceptible to any treatment. The continuous expansion of MRSA is still ongoing. The main goal of this article is to improve reading comprehension of MRSA by studying the prominent classes of antibiotics and their mechanism of resistance which are now susceptible or getting susceptible to the MRSA.

Detection of Novel spa Types in Clinical Isolates of Methicillin-Resistant Staphylococcus aureus in Iraq

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a gram-positive bacterium that is an opportunistic pathogen, causing infections in hospital settings and communities. MRSA has become a significant and increasing problem in Iraq. The aim of this study was to evaluate the genetic mutations of MRSA strains, especially in the spa gene, isolated from patients in Wassit, Iraq.Methods and Results: Biochemical tests were conducted to identify S. aureus isolates and then on MRSA. The MRSA was identified by the Chrome Agar method and confirmed by PCR with genotyping of the mecA gene. The disk diffusion method was used to detect antibiotic resistance to three different common antibiotics used at Wassit hospitals. The Vitek-2 compact system was utilized for the detection of the minimum inhibitory concentration (MIC) of vancomycin. All MRSA strains in this study were tested to screen the mecA gene, with 21 strains subjected to the molecular typing method for the spa gene. Out of 166 samples, 132(79.5%) contained S. aureus and 34(20.4%) were identified as MRSA. Genotyping showed that out of 34 MRSA, 31(91.2%) isolates were mecA-positive. The spa gene was detected in 21(61.8%) isolates out of 34 MRSA samples. The spa typing of 21 MRSA samples revealed four different spa types, as follows: t386 (3/14.3%), t3576 (1/4.8%), t10002 (1/4.8%), and t10234 (1/4.8%). High polymorphism rates were shown in isolates of spa type t386. Conclusion: Our data represent the first report to detect novel mutations in the spa gene in the MRSA clinical isolates from Wassit hospitals, Iraq.

Antimicrobial Resistance and Sports: The Scope of the Problem, Implications for Athletes' Health and Avenues for Collaborative Public Health Action.

Antimicrobial resistance (AMR) poses a global threat, leading to increased mortality and necessitating urgent action-however, its impact on athletes and the world of sports has hitherto been neglected. Sports environments (including athletic and aquatic) exhibit high levels of microbial contamination, potentially contributing to the spread of resistant microorganisms during physical activities. Moreover, the literature suggests that travel for sports events may lead to changes in athletes' gut microbiomes and potentially impact their antibiotic resistance profiles, raising questions about the broader implications for individual and public/global health. The prevalence of Staphylococcus aureus (S. aureus) among athletes (particularly those engaged in contact or collision sports) ranges between 22.4% and 68.6%, with MRSA strains being isolated in up to 34.9% of tested individuals. Factors such as training frequency, equipment sharing, delayed post-training showers, and a history of certain medical conditions are linked to higher colonization rates. Moreover, MRSA outbreaks have been documented in sports teams previously, highlighting the importance of implementing preventive measures and hygiene protocols in athletic settings. In light of the growing threat of AMR, there is a critical need for evidence-based treatment guidelines tailored to athletes' unique physiological demands to ensure responsible antibiotic use and mitigate potential health risks. While various initiatives-such as incorporating AMR awareness into major sporting events-aim to leverage the broad audience of sports to communicate the importance of addressing AMR, proactive measures (including improved AMR surveillance during large sporting events) will be indispensable for enhancing preparedness and safeguarding both athletes' and the general public's health. This narrative review thoroughly assesses the existing literature on AMR and antibiotic usage in the context of sports, aiming to illuminate areas where information may be lacking and underscoring the significance of promoting global awareness about AMR through sports.

Detection of mecA Genes in Hospital-Acquired MRSA and SOSA Strains Associated with Biofilm Formation.

Methicillin-resistant (MR) Staphylococcus aureus (SA) and others, except for Staphylococcus aureus (SOSA), are common in healthcare-associated infections. SOSA encompass largely coagulase-negative staphylococci, including coagulase-positive staphylococcal species. Biofilm formation is encoded by the icaADBC operon and is involved in virulence. mecA encodes an additional penicillin-binding protein (PBP), PBP2a, that avoids the arrival of β-lactams at the target, found in the staphylococcal cassette chromosome mec (SCCmec). This work aims to detect mecA, the bap gene, the icaADBC operon, and types of SCCmec associated to biofilm in MRSA and SOSA strains. A total of 46% (37/80) of the strains were S. aureus, 44% (35/80) S. epidermidis, 5% (4/80) S. haemolyticus, 2.5% (2/80) S. hominis, 1.25% (1/80) S. intermedius, and 1.25% (1/80) S. saprophyticus. A total of 85% were MR, of which 95.5% showed mecA and 86.7% β-lactamase producers; thus, Staphylococcus may have more than one resistance mechanism. Healthcare-associated infection strains codified type I-III genes of SCCmec; types IV and V were associated to community-acquired strains (CA). Type II prevailed in MRSA mecA strains and type II and III in MRSOSA (methicillin-resistant staphylococci other than Staphylococcus aureus). The operon icaADBC was found in 24% of SA and 14% of SOSA; probably the arrangement of the operon, fork formation, and mutations influenced the variation. Methicillin resistance was mainly mediated by the mecA gene; however, there may be other mechanisms that also participate, since biofilm production is related to genes of the icaADBC operon and methicillin resistance was not associated with biofilm production. Therefore, it is necessary to strengthen surveillance to prevent the spread of these outbreaks both in the nosocomial environment and in the community.

Unravelling the complex interplay between antibiotic consumption and adaptive changes in methicillin-resistant Staphylococcus aureus.

This study aims to elucidate the genomic dynamics driving the emergence of antimicrobial resistance (AMR), with a specific focus on the interplay between AMR and antimicrobial usage. We conducted a comprehensive analysis using a ST239 methicillin-resistant Staphylococcus aureus (MRSA) dataset over a continuous 12-year period from a single hospital. Genomic analyses were performed tracking the changes in MRSA populations, particularly the emergence of reduced vancomycin susceptibility, and assessing the impact of glycopeptide use on these emergence events. Our findings reveal a significant correlation between hospital glycopeptide usage and the selection of MRSA strains with reduced vancomycin susceptibility. Genomic analyses provided insights into the molecular mechanisms driving resistance emergence, including the slowing of the molecular clock rate in response to heightened antimicrobial consumption. In conclusion, this study the highlights the complex dynamics between AMR and antimicrobial use at the hospital level. The observed correlation between antimicrobial consumption and the development of less susceptible MRSA strains underscores the importance of antimicrobial stewardship programmes and the establishment of optimal consumption thresholds for mitigating AMR effectively.

Synergistic bactericidal effects of phage-enhanced antibiotic therapy against MRSA biofilms.

The prevalence of biofilm-associated medical device infections caused by methicillin-resistant Staphylococcus aureus (MRSA) presents a pressing medical challenge. The latest research demonstrates the potential of phage-antibiotic combinations (PACs) as a promising solution, notably in vitro antibiofilm efficacy. By adopting modified checkerboard and 24-h time-kill assays, the study investigated the synergistic action of phages combined with humanized-simulated doses of daptomycin (DAP) and ceftaroline (CPT). The results were promising: a combination of DAP, CPT, and either a 2-phage or 3-phage cocktail effectively exhibited bactericidal activity against both DAP non-susceptible vancomycin intermediate S. aureus MRSA and DAP-susceptible MRSA strains within 168-h biofilm models. Moreover, post-treatment evaluations revealed no discernible rise in antibiotic resistance or complete phage resistance. This pioneering work suggests the potential of PACs in addressing MRSA biofilm infections, setting the stage for further expansive research tailored to diverse bacterial strains and treatment durations.

Isolation and encapsulation of bacteriophage with chitosan nanoparticles for biocontrol of multidrug-resistant methicillin-resistant Staphylococcus aureus isolated from broiler poultry farms

This study was divided into two parts. The first part, the determination of methicillin-resistant Staphylococcus aureus (MRSA) prevalence in 25 broiler chicken farms, with the detection of multidrug resistant MRSA strains. The prevalence of MRSA was 31.8% (159 out of 500 samples) at the level of birds and it was 27% (27 out of 100) in the environmental samples. The highest antimicrobial resistance of the recovered MRSA strains was recorded to streptomycin (96%). All isolates (100%) had multidrug resistance (MDR) to four or more antibiotics with 16 distinct antibiotic resistant patterns, and multiple antibiotic resistance index (MARI) of 0.4–1. The second part, implementing novel biocontrol method for the isolated multidrug resistant MRSA strains through the isolation of its specific phage and detection of its survival rate at different pH and temperature degrees and lytic activity with and without encapsulation by chitosan nanoparticles (CS-NPs). Encapsulated and non-encapsulated MRSA phages were characterized using transmission electron microscope (TEM). Encapsulation of MRSA phage with CS-NPs increasing its lytic activity and its resistance to adverse conditions from pH and temperature. The findings of this study suggested that CS-NPs act as a protective barrier for MRSA phage for the control of multidrug resistant MRSA in broiler chicken farms.

Genotyping of Methicillin Resistant Staphylococcus aureus Clinical Isolates in Sulaimani City using Pulsed -Field Gel Electrophoresis

Methicillin-resistant Staphylococcus aureus (MRSA) strains cause serious nosocomial infections. The objective of this study was to detect and find the epidemiological relatedness among MRSA isolates which was collected in Emergency burn-and-plastic surgery hospital in Sulaimani city using Pulsed-filed gel electrophoresis (PFGE). Routine methods were used for detection of Staphylococcus aureus, further molecular confirmation was done by PCR assays targeting 16S rRNA gene. The cefoxitin-disc diffusion assay was used for detection MRSA, which is subsequently confirmed by the presence of the mecA gene. PFGE was chosen to observe the genetic relatedness of 10 MRSA strains using SmaI restriction endonuclease. All the isolates (100%, 50/50) carried 16S rRNA gene specific for S.aureus and (42%, 21/50) exhibited resistance to cefoxitin and harboured the mecA gene, thereby being classified as MRSA. Among 10 MRSA isolates, the PFGE pattern were grouped into four clusters. Three isolates belonging to cluster (C3) were indistinguishable, 4 isolates belonging to cluster (C4) were closely related, 2 isolates of the second cluster (C2) were possibly related and the first cluster (C1) with 1 MRSA isolate was unrelated. This finding shows 30% of the samples had indistinguishable fingerprints with no changes in banding profiles, while 40% of the samples had closely related patterns. PFGE is a good discriminatory tool for typing MRSA strains.

Evolutionary dynamics of the novel ST22-PT methicillin-resistant Staphylococcus aureus clone co-harbouring Panton–Valentine leucocidin and duplicated toxic shock syndrome toxin 1 genes

ObjectivesGlobally, the isolation of community-associated methicillin-resistant Staphylococcus aureus (MRSA) harbouring both the Panton–Valentine leucocidin (PVL) and toxic shock syndrome toxin 1 (TSST-1) genes is rare. However, we encountered an outbreak of the ST22-PT clone exhibiting this phenotype in Japan. Notably, the TSST-1 gene was duplicated in most of the strains. This study aimed to elucidate the mechanisms underlying this gene duplication. MethodsA total of 90 MRSA isolates were collected from the skin of outpatients in Fukuoka City, Japan, between 2017 and 2019. Whole-genome sequencing was performed on MRSA strains that were PVL and TSST-1 positive. ResultsA total of 43 (47.8%) strains produced TSST-1, 20 (22.2%) produced PVL, and 16 (17.8%) produced both. Fifteen isolates were classified as ST22/SCCmec type IVa (ST22-PT clone) and one as ST1/SCCmec type V (ST1-PT clone). Three distinct ST22-PT clones were identified: Fukuoka clone I (one PVL gene and one TSST-1 gene), Fukuoka clone II (addition of a TSST-1 gene to Fukuoka clone I), and Fukuoka clone III (marked by a chromosomal inversion in a large region from Fukuoka clone II). DiscussionFukuoka clone I may have integrated a novel pathogenicity island bearing the TSST-1 gene, leading to the emergence of Fukuoka clone II with a duplicated TSST-1 gene. This duplication subsequently instigated a chromosomal inversion in a large region owing to the homologous sequence surrounding TSST-1, giving rise to Fukuoka clone III. These findings provide crucial insights into the genetic evolution of MRSA.

The Prevalence and Antimicrobial Susceptibility of Methicillin-Resistant Staphylococcus aureus Before and After the COVID-19 Pandemic in a Tertiary Saudi Hospital.

Methicillin-resistant Staphylococcusaureus (MRSA) has become a major public health problem all over the world. After the 2019 coronavirus illness (COVID-19), the pandemic may have influenced research priorities and resource allocation, potentially affecting the ability to monitor MRSA trends. The study aimed to evaluate the prevalence of S. aureus, including MRSA infections, and their antimicrobial susceptibilities over the years 2019 and 2020 in a tertiary hospital in Makkah City, KSA. A total of 2128 and 1515 laboratory (lab) samples were collected during the years 2019 and 2020, respectively. From these samples, the prevalence of S. aureus, including MRSA, and their antibiotic susceptibility were identified using standard, automated, and molecular microbiological methods. The present study shows that the lab prevalence ofall S. aureus during 2019 was found to be 35.5%, of which MRSA was 44.8%. During 2020, the frequency of S. aureus strains was 16%, of which MRSA was 41.2%. The most common MRSA isolated during both years were colonizing pus swabs and urine samples. The results showed that MRSA susceptibility against antimicrobial agents in 2019 was as follows: vancomycin (100%), linezolid (100%), trimethoprim-sulfamethoxazole (88%), and doxycycline (34.2%). The MRSA strains isolated during 2020 were as follows: vancomycin (100%), linezolid (96%), trimethoprim-sulfamethoxazole (100%), and doxycycline (24.3%). There was no significant difference in the incidence and antimicrobial resistance rates of MRSA over the two years. It was concluded that the prevalence rates of MRSA have not increased in 2020 when compared to 2019. Vancomycin, linezolid, trimethoprim-sulfamethoxazole, and doxycycline remain susceptible to the positive collected MRSA strains. There was no significant difference between the prevalence and antimicrobial resistance rates of MRSA between 2019 and 2020. Continued research efforts are needed to address this persistent public health threat. Strategies to control the spread of MRSA should include early detection of MRSA and surveillance, even during pandemics.

Assessment of minimum inhibitory concentration to vancomycin, tigecycline, linezolid, daptomycin, ceftaroline and mupirocin against methicillin resistant Staphylococcus aureus clinical isolates by antibiotic gradient strips

Background: Staphylococcus aureus infections are one of the most common and serious hospital-acquired infections seen in developing countries. Methicillin resistant Staphylococcus aureus (MRSA) is an important human pathogen and normally colonized in body parts including skin, nose, perineum and throat. MRSA is resistant not only to all β-lactam groups but also other antibiotics including aminoglycosides, tetracycline and macrolides. In the present study the efficacy of agents used in the management of MRSA infections was determined by antibiotic gradient testing. Methods: A total of 60 clinical isolates of MRSA strains were collected from various diagnostic labs in central Kerala. Clinical isolates were reconfirmed as MRSA by gram staining, yellow-coloured colonies on mannitol salt Agar (MSA). Antibiotic susceptibility testing was done by disc diffusion method as recommended by CLSI guidelines. S. aureus isolates resistant to cefoxitin (30 µg) was identified as MRSA. Antibiotic gradient testing was performed to determine the MIC of vancomycin, tigecycline, linezolid, daptomycin, ceftaroline and mupirocin against MRSA isolates. Results: All the 60 MRSA isolates tested were sensitive to vancomycin, tigecycline, linezolid, daptomycin, ceftaroline and mupirocin (100%) and none of the MRSA isolates show resistance. Conclusions: Results of present study indicates that these agents may be used alongside vancomycin in management of infection caused by MRSA.

Prevalence of Panton-Valentine leucocidin (pvl) and exfoliative toxin A (eta) gene within methicillin resistant and susceptible Staphylococcus aureus in an urban tertiary hospital: A molecular epidemiology pilot study

Background Staphylococcus aureus is well known to cause a multitude of clinical manifestations, from mild to severe bloodstream infections that could lead to death. Infections are common, either in community-acquired or hospital-acquired settings, and treatment remains a challenge due to methicillin-resistant Staphylococcus aureus (MRSA). The pathogenesis of S. aureus is mediated by several cell-surface and secreted virulence factors. The virulence factors discussed in this study are Panton-Valentine leucocidin (pvl) and exfoliative toxin A (eta). Identifying both pvl and eta gene may help in studying bacterial pathogenesis and biology thus creating possible therapeutic pathway or intervention. Our pilot study aimed to observe pvl and eta as virulence gene prevalence in a North Sumatera tertiary referral health center. Methods Our study was a descriptive-analytical observational study with a cross-sectional design in which we collected isolates over a single time period. The frequency of genes is reported as a percentage comparison between MRSA and methicillin-susceptible S. aureus (MSSA). Qualitative gene prevalence analysis was carried out using the polymerase chain reaction (PCR). Results Our results showed that from 38 MRSA sample isolates, 32 samples were found to be pvl-positive, or 84,3% of the total samples. From 40 MSSA sample isolates, one sample was found to be pvl-positive MSSA, or 97,5%. Regarding eta, from 38 MRSA sample isolates, 81,6% of the total sample did not have eta, while from 40 MSSA sample isolates, all samples were found to be positive for eta. We found that both pvl and eta were significantly more likely to be expressed in the MSSA strain. Conclusions Our study shows that pvl and eta are more likely expressed in MSSA strains than in MRSA strains in Indonesia.

Comparative effectiveness of daptomycin versus vancomycin among patients with methicillin-resistant Staphylococcus aureus (MRSA) bloodstream infections: A systematic literature review and meta-analysis.

In the treatment of methicillin-resistant Staphylococcus aureus (MRSA) bloodstream infections (BSIs), vancomycin stands as the prevalent therapeutic agent. Daptomycin remains an alternative antibiotic to treat MRSA BSIs in cases where vancomycin proves ineffective. However, studies have conflicted on whether daptomycin is more effective than vancomycin among patients with MRSA BSI. To compare the effectiveness of daptomycin and vancomycin for the prevention of mortality among adult patients with MRSA BSI. Systematic searches of databases were performed, including Embase, PubMed, Web of Science, and Cochrane Library. The Newcastle Ottawa Scale (NOS) and Revised Cochrane risk-of-bias tool for randomized trials (RoB 2) were used to assess the quality of individual observational and randomized control studies, respectively. Pooled odd ratios were calculated using random effects models. Twenty studies were included based on a priori set inclusion and exclusion criteria. Daptomycin treatment was associated with non-significant lower mortality odds, compared to vancomycin treatment (OR = 0.81; 95% CI, 0.62, 1.06). Sub-analyses based on the time patients were switched from another anti-MRSA treatment to daptomycin demonstrated that switching to daptomycin within 3 or 5 days was significantly associated with 55% and 45% decreased odds of all-cause mortality, respectively. However, switching to daptomycin any time after five days of treatment was not significantly associated with lower odds of mortality. Stratified analysis based on vancomycin minimum inhibitory concentration (MIC) revealed that daptomycin treatment among patients infected with MRSA strains with MIC≥1 mg/L was significantly associated with 40% lower odds of mortality compared to vancomycin treatment. Compared with vancomycin, an early switch from vancomycin to daptomycin was significantly associated with lower odds of mortality. In contrast, switching to daptomycin at any time only showed a trend towards reduced mortality, with a non-significant association. Therefore, the efficacy of early daptomycin use over vancomycin against mortality among MRSA BSIs patients may add evidence to the existing literature in support of switching to daptomycin early over remaining on vancomycin. More randomized and prospective studies are needed to assess this association.

Phage-antibiotic synergy against daptomycin-nonsusceptible MRSA in an ex vivo simulated endocardial pharmacokinetic/pharmacodynamic model.

Phage-antibiotic combinations (PAC) offer a potential solution for treating refractory daptomycin-nonsusceptible (DNS) methicillin-resistant Staphylococcus aureus (MRSA) infections. We examined PAC activity against two well-characterized DNS MRSA strains (C4 and C37) in vitro and ex vivo. PACs comprising daptomycin (DAP) ± ceftaroline (CPT) and a two-phage cocktail (Intesti13 + Sb-1) were evaluated for phage-antibiotic synergy (PAS) against high MRSA inoculum (109 CFU/mL) using (i) modified checkerboards (CB), (ii) 24-h time-kill assays (TKA), and (iii) 168-h ex vivo simulated endocardial vegetation (SEV) models. PAS was defined as a fractional inhibitory concentration ≤0.5 in CB minimum inhibitory concentration (MIC) or a ≥2 log10 CFU/mL reduction compared to the next best regimen in time-kill assays and SEV models. Significant differences between regimens were assessed by analysis of variance with Tukey's post hoc modification (α = 0.05). CB assays revealed PAS with Intesti13 + Sb-1 + DAP ± CPT. In 24-h time-kill assays against C4, Intesti13 + Sb-1 + DAP ± CPT demonstrated synergistic activity (-Δ7.21 and -Δ7.39 log10 CFU/mL, respectively) (P < 0.05 each). Against C37, Intesti13 + Sb-1 + CPT ± DAP was equally effective (-Δ7.14 log10 CFU/mL each) and not significantly different from DAP + Intesti13 + Sb-1 (-Δ6.65 log10 CFU/mL). In 168-h SEV models against C4 and C37, DAP ± CPT + the phage cocktail exerted synergistic activities, significantly reducing bio-burdens to the detection limit [2 log10 CFU/g (-Δ7.07 and -Δ7.11 log10 CFU/g, respectively)] (P < 0.001). At 168 h, both models maintained stable MICs, and no treatment-emergent phage resistance occurred with DAP or DAP + CPT regimens. The two-phage cocktail demonstrated synergistic activity against two DNS MRSA isolates in combination with DAP + CPT in vitro and ex vivo. Further in vivo PAC investigations are needed.

Staphylococcal scalded skin syndrome in neonate: Another face of CA-MRSA.

Staphylococcal scalded skin syndrome (SSSS), also known as Ritter's disease, in its severe form occurs predominantly in infants and children. It is caused by infection with group II (often phage group 71) Staphylococcus aureus. The foci of infection include nasopharynx, less commonly umbilicus, urinary tract, superficial abrasion, conjunctivae, and blood. Staphylococci are non-motile, non-spore-forming, catalase-positive, gram-positive cocci that appear predominantly as grape-like clusters. Although this organism is frequently a part of normal human microbial flora, it can cause significant opportunistic infections under certain conditions such as when extremes of age groups are involved, the presence of indwelling medical devices, and intravenous (iv) drug abuse. Staphylococcus aureus may cause a variety of infectious manifestations ranging from relatively benign skin infections to life-threatening systemic illnesses. SSSS caused by S. aureus strains produces exfoliative toxins which result in the development of blisters, erythema, and desquamation. Here, we present a case of an 11-day-old neonate who was diagnosed with SSSS. The causative agent responsible for this syndrome was identified as methicillin-resistant Staphylococcus aureus (MRSA). The molecular characterization of the gene Panton-Valentine leukocidin (PVL) was done by polymerase chain reaction (PCR) and was detected positive for PVL which is a distinctive virulence factor seen almost in all of the community-acquired MRSA strains. The patient was discharged after parenteral clindamycin therapy with almost complete resolution of symptoms.

Factors associated with foreign body infection in methicillin-resistant Staphylococcus aureus bacteremia.

We aimed to compare patient characteristics, MRSA sequence types, and biofilm production of MRSA strains that did and did not cause a foreign body infection in patients with MRSA bloodstream infections (BSI). All adult patients with MRSA BSI hospitalized in two hospitals were identified by clinical microbiology laboratory surveillance. Only patients who had at least one implanted foreign body during the episode of BSI were included. In July 2018 - March 2022, of 423 patients identified with MRSA BSI, 118 (28%) had ≥1 foreign body. Among them, 51 (43%) had one or more foreign body infections. In multivariable analysis, factors associated with foreign body infection were history of MRSA infection in the last year (OR=4.7 [1.4-15.5], p=0.012) community-associated BSI (OR=68.1 [4.2-1114.3], p=0.003); surgical site infection as source of infection (OR=11.8 [2-70.4], p=0.007); presence of more than one foreign body (OR=3.4 [1.1-10.7], p=0.033); interval between foreign body implantation and infection <18 months (OR=3.3 [1.1-10], p=0.031); and positive blood culture ≥48h (OR=16.7 [4.3-65.7], p<0.001). The most prevalent sequence type was ST8 (39%), followed by ST5 (29%), and ST105 (20%) with no significant difference between patients with or without foreign body infection. Only 39% of MRSA isolates formed a moderate/strong biofilm. No significant difference was observed between patients with foreign body infection and those without foreign body infection. In multivariable analysis, subjects infected with a MRSA isolate producing moderate/strong in vitro biofilm were more likely to have a history of MRSA infection in the last year (OR=3.41 [1.23-9.43]), interval between foreign body implantation and MRSA BSI <18 months (OR=3.1 [1.05-9.2]) and ST8 (OR=10.64 [2-57.3]). Most factors associated with foreign body infection in MRSA BSI were also characteristic of persistent infections. Biofilm-forming isolates were not associated with a higher risk of foreign-body infection but appeared to be associated with MRSA genetic lineage, especially ST8.

Co-colonization of methicillin-resistant Staphylococcus aureus and Candida spp. in children with malignancies

This study aimed to evaluate the interaction between methicillin-resistant Staphylococcus aureus(MRSA) and Candida spp. in the oral cavity of children with malignancies under chemotherapy. We evaluated the expression level of Als3p and mecA in Candida spp. and MRSA strains in both single colonization and co-colonization condition. Oral and nasal samples were collected by dry sponge swabs in 10 ml of sterile phosphate-buffered saline. The MRSA and Candida spp. was confirmed using the PCR method and mecA and Als3p genes, respectively. The SYBR Green-based quantitative real-time PCR was used to evaluate the relative expression levels of mecA and Als3p genes in MRSA and Candida spp., respectively. The frequency of S. aureus in oral-only and nasal-only swab samples were 14.1% (n = 24/170). 58.3% (n = 14/24) and 29.2% (n = 7/24) of S. aureus isolated from oral and nasal samples were MRSA, respectively. Among Candida species, C. albicans (n = 28/170; 16.5%) had the highest frequency. The oral co-colonization of MRSA and Candida spp. was detected in 4.7% (n = 8/170) patients. The overall average of gene expression levels among all Candida spp. and MRSA isolates indicated that the mecA and Als3p genes expression increased six and two times in co-colonization conditions compared to single colonization conditions, respectively. Our findings revealed the importance of polymicrobial infection in clinical settings and stated that it is possible that Candida spp. facilitates the infection of S. aureus and can lead to systemic infection in co-colonized patients.

Sensitizing methicillin-resistant Staphylococcus aureus (MRSA) to cefuroxime: the synergic effect of bicarbonate and the wall teichoic acid inhibitor ticlopidine.

Methicillin-resistant Staphylococcus aureus (MRSA) strains are a major challenge for clinicians due, in part, to their resistance to most β-lactams, the first-line treatment for methicillin-susceptible S. aureus. A phenotype termed "NaHCO3-responsiveness" has been identified, wherein many clinical MRSA isolates are rendered susceptible to standard-of-care β-lactams in the presence of physiologically relevant concentrations of NaHCO3, in vitro and ex vivo; moreover, such "NaHCO3-responsive" isolates can be effectively cleared by β-lactams from target tissues in experimental infective endocarditis (IE). One mechanistic impact of NaHCO3 exposure on NaHCO3-responsive MRSA is to repress WTA synthesis. This NaHCO3 effect mimics the phenotype of tarO-deficient MRSA, including sensitization to the PBP2-targeting β-lactam, cefuroxime (CFX). Herein, we further investigated the impacts of NaHCO3 exposure on CFX susceptibility in the presence and absence of a WTA synthesis inhibitor, ticlopidine (TCP), in a collection of clinical MRSA isolates from skin and soft tissue infections (SSTI) and bloodstream infections (BSI). NaHCO3 and/or TCP enhanced susceptibility to CFX in vitro, by both minimum inhibitor concentration (MIC) and time-kill assays, as well as in an ex vivo simulated endocarditis vegetations (SEV) model, in NaHCO3-responsive MRSA. Furthermore, in experimental IE (presumably in the presence of endogenous NaHCO3), pre-exposure to TCP prior to infection sensitized the NaHCO3-responsive MRSA strain (but not the non-responsive strain) to enhanced clearances by CFX in target tissues. These data support the notion that NaHCO3 is acting similarly to WTA synthesis inhibitors, and that such inhibitors have potential translational applications in the treatment of certain MRSA strains in conjunction with specific β-lactam agents.

Susceptibility to postbiotics - enterocins of methicillin-resistant Staphylococcus aureus strains isolated from rabbits

There is a major problem with the rising occurrence of highly virulent and multiply-resistant strains, including methicillin-resistant Staphylococcus aureus (MRSA), because of their difficult treatment. This study aimed to evaluate the antibacterial and antibiofilm effect of new enterocins (Ent) against potential pathogenic MRSA strains isolated from rabbits. Staphylococci were identified with PCR and screened for methicillin/oxacillin/cefoxitin resistance (MR) using the disk diffusion method and the PBP2’ Latex Agglutination Test Kit. Enzyme production, hemolysis, DNase activity, slime production, and biofilm formation were tested in MRSA strains. The susceptibility of MRSA to eight partially-purified enterocins (Ent) produced by E. faecium and E. durans strains was checked using agar spot tests. The antibiofilm activity of Ents was tested using a quantitative plate assay. Out of 14 MRSA, PBP testing confirmed MR in 8 strains. The majority of MRSA showed DNase activity and β-hemolysis. Slime production and moderate biofilm formation were observed in all strains. MRSA were susceptible to tested Ents (100–12,800 AU/mL; except Ent4231). The antibiofilm effect of Ents (except Ent4231) was noted in the high range (64.9–97.0%). These results indicate that enterocins offer a promising option for the prevention and treatment of bacterial infections caused by biofilm-forming MRSA.

Water extracts of pomegranate peel and hojicha tea leaf inhibit the growth of methicillin-resistant staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) strains capable of causing infections and food poisoning have emerged worldwide in response to the use of various antimicrobial agents. To date, several drugs have been used to combat these bacteria. We focused herein on functional foods (particularly food waste products), which are safer and exhibit fewer side effects than drugs. The objective of present study is to elucidate the growth inhibitory effects of several food waste products on Staphylococcus aureus using three different Staphylococcus aureus strains, JCM2413 as the standard strain, and MW2 and N315 as MRSA strains. One percent water extracts of ten food waste products (grapefruit peel, pineapple leaf, dragon fruit peel, persimmon peel, turnip leaf, pomegranate peel, apple peel, potato peel, hojicha tea leaf, and banana peel) were added to the medium, and colonies were counted after a 24-hour incubation. Significant decreases were observed in MW2 and N315 colonies with pomegranate peel and hojicha tea leaf extracts. By contrast, no significant difference was observed in JCM2413 in any of the extracts. We demonstrated that water extracts of pomegranate peel and hojicha tea leaves possess bacteriostatic activity specific to MRSA. Our findings help to improve food safety with reducing food loss. However, further studies are needed to elucidate the detailed mechanism of the effects of pomegranate peel and hojicha tea on MRSA.