Methicillin-resistant Staphylococcus Aureus Isolates Research Articles

Methicillin-resistant Staphylococcus aureus outbreak in a Dutch equine referral clinic.

In 2020 and 2022, nine cases of surgical site infections with a methicillin-resistant Staphylococcus aureus (MRSA) were diagnosed in horses in an equine referral clinic. Sixteen isolates (horses, n=9; environment, n=3; and staff members, n=4) were analysed retrospectively using Nanopore whole-genome sequencing to investigate the relatedness of two suspected MRSA outbreaks (2020 and 2022). The MRSA isolates belonged to ST398 and ST612. ST398 genomes from 2020 and 2022 formed three phylogenetic clusters. The first ST398 cluster from 2020 consisted of isolates from five horses and one staff member, and we suspected within clinic transmission. The second cluster of ST398 isolates from 2022 originated from two horses and two staff members but showed higher single nucleotide polymorphism (SNP) distances. One ST398 isolate from an individual staff member was not related to the other two clusters. The ST612 isolates were isolated in 2022 from two horses and three environmental samples and showed very low SNP distances (<7 SNPs), indicating the transmission of MRSA ST612 in this clinic in 2022. Molecular characterization revealed an abundant set of virulence genes and plasmids in the ST612 isolates in comparison to ST398 isolates. Phenotypic antimicrobial susceptibility showed that differences between the two sequence types were consistent with the genetic characteristics. MRSA ST612 has not been reported in Europe before, but it is a dominant clone in African hospitals and has been described in horses and people working with horses in Australia, indicating the importance of surveillance.

Panton-Valentine leucocidin gene in methicillin resistant Staphylococcus aureus isolated from tertiary care hospital in Nepal.

Methicillin-resistant Staphylococcus aureus (MRSA) expresses the Panton-Valentine leukocidin (PVL) virulence gene, which is associated with community and hospital-acquired severe MRSA infections. The objective of this study was to determine the prevalence and antibiotic susceptibility profile with a focus on the presence of the PVL gene among MRSA isolates in healthcare settings. A total of 1,207 clinical specimens and 304 hospital environment swabs were collected in a tertiary care hospital in Nepal, and investigated following basic microbiological techniques. S. aureus was confirmed with the coagulase test. An antibiotic susceptibility test (AST) was performed by the Kirby-Bauer method and screening for MRSA was carried out by the cefoxitin disc diffusion method guided by the Clinical and Laboratory Standards Institute (CLSI), 2020. DNA was extracted and used in a polymerase chain reaction (PCR) to detect mecA and PVL genes. Of the 1,511 samples, 45 (2.9%) S. aureus (23 clinical and 22 environmental) were isolated. Among them, 69.6% (16/23) and 27.3% (6/22) were MRSA in clinical and environmental isolates, respectively. Twelve (52.2%) clinical isolates and seven (31.8%) environmental isolates were multidrug resistant. The majority of isolates were susceptible to vancomycin and linezolid. The PVL gene was detected in 18.2% (n = 4/22) of the MRSA isolates, of which three were from clinical sources and one was from an environmental swab. The prevalence of MRSA, and PVL-producing S. aureus were higher in the hospital setting. Hence, immediate and urgent implementation of infection control and sanitation measures are needed in the hospital.

Methicillin-Resistant Staphylococcus aureus (MRSA): Prevalence and Antimicrobial Susceptibility Profile among Patients Attending Tertiary Care Hospital in Northern India

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important causative agents of hospital and community associated bacterial infections. The increasing rate of prevalence indicates the need to study the antimicrobial susceptibility profile patterns. This study was aimed to determine the prevalence and antibiotic susceptibility profile pattern of methicillin-resistant Staphylococcus aureus (MRSA) isolates among patients attending a tertiary care hospital in northern India. 150 clinical isolates of S. aureus were included in the study. Kirby–Bauer disc diffusion method was performed to identify antibiotic susceptibility profile, phenotypically methicillin-resistant Staphylococcus aureus (MRSA) was identified by using cefoxitin disc (30 μg). Characteristics of the isolates in this study were determined by traditional biochemical tests including catalase, coagulase, and test for DNAse, and fermentation of mannitol. Antibiotic susceptibility tests were carried out by disc agar diffusion test. Staphylococcus aureus ATCC 25923 was used as a reference control organism. Out of 150 clinical isolates of S. aureus, 97 (64.67%) were methicillin-resistant Staphylococcus aureus (MRSA), and 53 (35.33%) were methicillin-sensitive Staphylococcus aureus (MSSA). 21-30 years age group has the highest percentage of the isolates (28.00%) and the age group with lowest percentage was found in the age groups 71-80 years. The source of isolates with the highest frequency was pus (56.67%), followed by urine (20.00%), and the source with the least number of isolates was serous fluid (0.67%). Teicoplanin, Linezolid, Vancomycin, Gentamicin and Tetracycline were found to be the most effective antibiotics against MRSA isolates. The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) was 64.67%. Teicoplanin, Linezolid, Vancomycin, Gentamicin and Tetracycline found to be the most effective antibiotics against the MRSA isolates. Thus, it is believed that these antibiotics can be used in the treatment of infections caused by methicillin-resistant Staphylococcus aureus (MRSA) in the study area.

Determination of in vitro synergy and antibiofilm activities of antimicrobials and essential oil components

Using existing adrentimicrobials with essential oil components to prevent antimicrobial resistance is an alternative strategy. This study aimed to evaluate the resistance status, synergistic combinations, and in vitro biofilm formation activities of clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA), Stenotrophomonas maltophilia and Candida albicans against antimicrobial agents and cinnamaldehyde, carvacrol, eugenol, limonene and eucalyptol. Antimicrobial activities were evaluated by microdilution, cytotoxicity by XTT, synergy by checkerboard and time-kill, and biofilm inhibition by microplate methods. Cinnamaldehyde and carvacrol showed strong antimicrobial activity. Synergistic effects were observed when using all essential oils with antimicrobials. Only two C. albicans isolates showed antagonism with cinnamaldehyde and fluconazole. The constituents showed cytotoxic effects in the L929 cell line (except limonene). A time-kill analysis revealed a bacteriostatic effect on S. maltophilia and MRSA isolates and a fungicidal effect on C. albicans isolates. These results are important for further research to improve antimicrobial efficacy or to develop new agents.

Isolation of community-associated methicillin-resistant Staphylococcus aureus (MRSA) sequence type (ST) 30 from house rats (Rattus tanezumi) in Hong Kong.

Methicillin-resistant Staphylococcus aureus (MRSA) is of major public health concern due to its resistance to multiple antibiotics. This resistance has been observed in various settings, including hospitals and communities, and has been detected in both animals and humans. Although peridomestic rat species (Rattus spp.) are well described reservoirs of several human pathogens and antimicrobial resistant bacteria, little is known about their role in MRSA epidemiology. In order to investigate whether Rattus spp. in Hong Kong are potential carriers of MRSA, 221 rats were caught from various ecological areas and nasopharyngeal samples were cultured on MRSA selective media. Genotypic characteristics of MRSA were confirmed by whole genome sequencing. Two clonal sequence type (ST) 30 MRSA isolates, harbouring mecA on staphylococcal chromosome cassette (SCC) mec type IVc, were cultured from two house rats (Rattus tanezumi) caught in two densely populated urban areas. To the best of the authors' knowledge, this is the first detection of community-associated (CA)-MRSA strain ST30 SCCmec IVc in peridomestic rodents in Hong Kong and globally. Our finding indicates that house rats can be carriers of MRSA strains that are widely distributed in the community.

New insights: mupirocin resistance among methicillin-resistant Staphylococcus aureus (MRSA) infection in southern Indian region

: Methicillin-resistant (MRSA) remains to be the most concerned pathogen with enhanced multi-drug resistance in clinical set-ups. With an immediate need to control MRSA infection, there is a steep rise in the use of mupirocin as a decolonization regime.: A total of 65 MRSA samples were analysed in the present study. infection identified by standard conventional phenotypic methods. The antibiotic susceptibility pattern of mupirocin (5 μg/ml) was found using Kirby Bauer’s disc diffusion method and the minimum inhibitory concentration (MIC) determined using MIC E-test strips of mupirocin.: All the bacterial strains presented with MIC value less than 4 μg/ml, 23 isolates falls in the range of 0.05 μg/ml and 0.19 μg/ml, 12 strains MIC ranged between 0.125 and 0.640 μg/ml. However, five MRSA isolates showed no zone against mupirocin E-strip.: In present study, 92% of isolates were sensitive to mupirocin, this gives hope that mupirocin can be promising in the elimination of MRSA strains, a major concern in clinical practice. Also, the use of 5µg mupirocin discs is economical as compared to mupirocin E-strips, mupirocin discs can be used in a low resourceful clinical setups. However, our study revealed no difference in sensitivity for both the E-strip and disc method.

A Comparative Analysis of MRSA: Epidemiology and Antibiotic Resistance in Greece and Romania.

This study provides a comparative analysis of 243 Methicillin-resistant Staphylococcus aureus (MRSA) isolated strains from Greece and Romania, focusing on their epidemiology and antibiotic resistance patterns. Laboratory procedures included phenotypic and automated identification methods, susceptibility testing, DNA isolation, and PCR for detecting antibiotic resistance genes (MecA, SCCmec). Our study results show significant regional differences. In both regions, males have higher MRSA infection rates than females, but the percentages vary. Greece has a higher incidence of MRSA in younger age groups compared to Romania. The majority of MRSA infections occur in inpatient settings in both countries, highlighting the necessity for enhanced infection control measures. Antibiotic resistance profiles reveal higher resistance to several antibiotics in Greece compared to Romania. A molecular analysis shows a widespread distribution of antibiotic resistance genes among MRSA isolates in Greece. These results highlight the necessity for accomplished preventive strategies and optimized treatment protocols.

Antibacterial and antibiofilm potentials of vancomycin-loaded niosomal drug delivery system against methicillin-resistant Staphylococcus aureus (MRSA) infections

The threat of methicillin-resistant Staphylococcus aureus (MRSA) is increasing worldwide, making it significantly necessary to discover a novel way of dealing with related infections. The quick spread of MRSA isolates among infected individuals has heightened public health concerns and significantly limited treatment options. Vancomycin (VAN) can be applied to treat severe MRSA infections, and the indiscriminate administration of this antimicrobial agent has caused several concerns in medical settings. Owing to several advantageous characteristics, a niosomal drug delivery system may increase the potential of loaded antimicrobial agents. This work aims to examine the antibacterial and anti-biofilm properties of VAN-niosome against MRSA clinical isolates with emphasis on cytotoxicity and stability studies. Furthermore, we aim to suggest an effective approach against MRSA infections by investigating the inhibitory effect of formulated niosome on the expression of the biofilm-associated gene (icaR). The thin-film hydration approach was used to prepare the niosome (Tween 60, Span 60, and cholesterol), and field emission scanning electron microscopy (FE-SEM), an in vitro drug release, dynamic light scattering (DLS), and entrapment efficiency (EE%) were used to investigate the physicochemical properties. The physical stability of VAN-niosome, including hydrodynamic size, polydispersity index (PDI), and EE%, was analyzed for a 30-day storage time at 4 °C and 25 °C. In addition, the human foreskin fibroblast (HFF) cell line was used to evaluate the cytotoxic effect of synthesized niosome. Moreover, minimum inhibitory and bactericidal concentrations (MICs/MBCs) were applied to assess the antibacterial properties of niosomal VAN formulation. Also, the antibiofilm potential of VAN-niosome was investigated by microtiter plate (MTP) and real-time PCR methods. The FE-SEM result revealed that synthesized VAN-niosome had a spherical morphology. The hydrodynamic size and PDI of VAN-niosome reported by the DLS method were 201.2 nm and 0.301, respectively. Also, the surface zeta charge of the prepared niosome was − 35.4 mV, and the EE% ranged between 58.9 and 62.5%. Moreover, in vitro release study revealed a sustained-release profile for synthesized niosomal formulation. Our study showed that VAN-niosome had acceptable stability during a 30-day storage time. Additionally, the VAN-niosome had stronger antibacterial and anti-biofilm properties against MRSA clinical isolates compared with free VAN. In conclusion, the result of our study demonstrated that niosomal VAN could be promising as a successful drug delivery system due to sustained drug release, negligible toxicity, and high encapsulation capacity. Also, the antibacterial and anti-biofilm studies showed the high capacity of VAN-niosome against MRSA clinical isolates. Furthermore, the results of real-time PCR exhibited that VAN-niosome could be proposed as a powerful strategy against MRSA biofilm via down-regulation of icaR gene expression.

Biofilm-producing ability of methicillin-resistant Staphylococcus aureus clinically isolated in China

BackgroundStaphylococcus aureus, a commensal bacterium, colonizes the skin and mucous membranes of approximately 30% of the human population. Apart from conventional resistance mechanisms, one of the pathogenic features of S. aureus is its ability to survive in a biofilm state on both biotic and abiotic surfaces. Due to this characteristic, S. aureus is a major cause of human infections, with Methicillin-Resistant Staphylococcus aureus (MRSA) being a significant contributor to both community-acquired and hospital-acquired infections.ResultsAnalyzing non-repetitive clinical isolates of MRSA collected from seven provinces and cities in China between 2014 and 2020, it was observed that 53.2% of the MRSA isolates exhibited varying degrees of ability to produce biofilm. The biofilm positivity rate was notably high in MRSA isolates from Guangdong, Jiangxi, and Hubei. The predominant MRSA strains collected in this study were of sequence types ST59, ST5, and ST239, with the biofilm-producing capability mainly distributed among moderate and weak biofilm producers within these ST types. Notably, certain sequence types, such as ST88, exhibited a high prevalence of strong biofilm-producing strains. The study found that SCCmec IV was the predominant type among biofilm-positive MRSA, followed by SCCmec II. Comparing strains with weak and strong biofilm production capabilities, the positive rates of the sdrD and sdrE were higher in strong biofilm producers. The genetic determinants ebp, icaA, icaB, icaC, icaD, icaR, and sdrE were associated with strong biofilm production in MRSA. Additionally, biofilm-negative MRSA isolates showed higher sensitivity rates to cefalotin (94.8%), daptomycin (94.5%), mupirocin (86.5%), teicoplanin (94.5%), fusidic acid (81.0%), and dalbavancin (94.5%) compared to biofilm-positive MRSA isolates. The biofilm positivity rate was consistently above 50% in all collected specimen types.ConclusionsMRSA strains with biofilm production capability warrant increased vigilance.

Molecular Detection of the mecA and some Virulence Determinants in Methicillin-Resistant Staphylococcus aureus

Background: Methicillin Resistant Staphylococcus aureus (MRSA) is globally acknowledged as a prominent contributor to both hospital-acquired and community infections. Understanding key virulence factors including coagulase production, hemolysis ability and biofilm formation, is crucial. Objective: The study aimed to establish a molecular characterization of mecA gene and virulence factors genes (hla, icaA, and coa) in clinical isolates of MRSA obtained from two hospitals in Baghdad. Materials and Methods: A hundred and five isolates were obtained from clinical sources from November 2022 to March 2023 and their antibiotic sensitivity was assessed using the agar diffusion test against seven different antibiotics (Azithromycin, Ciprofloxacin, Nitrofurantoin, Rifampin, Trimethoprim, Ofloxacin and Oxacillin), through Conventional Polymerase Chain Reaction, the presence of virulence factor genes including mecA, hla, icaA, and coa, was determined in MRSA isolates. Results: All MRSA isolates (100%) harbored the mecA, hla, and icaA genes while the coa gene was recognized in 50% of the isolates. Regarding antibiotic susceptibility, all MRSA isolates (100%) demonstrated sensitivity to Nitrofurantoin. Additionally, 96.8% of the isolates were sensitive to Oxacillin. Conclusion: Molecular detection of methicillin resistance genes and virulence genes can be used to diagnose MRSA isolates in hospitals. The presence of these genes may affect their pattern of sensitivity to antibiotics.

Assessing Mupirocin Resistance in MRSA Isolates in Hospitals in Cleveland, OH and Detroit, MI

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a common pathogen responsible for nosocomial and community-acquired infections with high morbidity and mortality1. MRSA nasal colonization is a major risk factor for developing infection in the hospital setting2,3. Decolonization of MRSA carriers is a strategy to decrease recurrence or to prevent new MRSA infections3,4. Decolonization with nasal mupirocin 2% and chlorhexidine baths has been shown to decrease the risk of MRSA infection after hospital discharge3. Mupirocin is an RNA synthetase inhibitor with activity against MRSA5. Resistance of MRSA isolates to mupirocin has been described previously6. As topical disinfectants play a crucial role in prevention of MRSA infection in a variety of settings, it is important to monitor the emergence of resistance. The goal of this study was to determine the prevalence of mupirocin resistance among MRSA samples isolated from two different regions in the United States (U.S). Methods: Our study had a total of 474 MRSA samples that were obtained from hospitals in Detroit, MI (287 samples) and Cleveland, OH (187 samples). After whole genome sequencing using NextSeq (Illumina Inc., CA) platform the data was analyzed using ResFinder 4.1, to identify antimicrobial resistance which can be either acquired or chromosomally mediated mutations. To visualize the presence of genes of interest the resistance genes were tallied on a spread sheet. Results: Mupirocin resistance gene was detected in five of 287 (1.74%) MRSA samples from the Detroit hospitals, all of which were associated with the mupA gene. Samples collected from the Cleveland area hospital demonstrated mupirocin resistance in seven samples of 187 (3.74%), all associated again with the mupA gene. One sample from the Detroit group showed resistance to both mupirocin and chlorhexidine. Conclusions: Prevalence of mupirocin resistance gene varied between the two hospital locations. Resistance to mupirocin has been documented in association with mutations in the mupA gene as well as chromosomal point mutations that can lead to either low or high-level resistance7,8. Although the mechanisms are not fully clear, mupA gene has been associated with high-level resistance9. Mupirocin resistance among MRSA isolates has increased over time9. MRSA infections remain an important etiology of nosocomial and community-acquired infections and common practice to combat this issue is universal decolonization with mupirocin10. It is critical to understand and monitor for development of mupirocin resistance as mupirocin remains one of the most effective tools to prevent invasive infection with MRSA in many patient populations.

Assessing chlorhexidine resistance in MRSA isolates from hospitals in Cleveland, OH and Detroit, MI

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common causes of procedure-related, skin, and soft-tissue infections. Hospitalized patients who are colonized with MRSA are at a higher risk of developing invasive infections after discharge. Chlorhexidine, an antiseptic/disinfectant, has been used to reduce carriage and prevent infections in these patients. Studies have shown chlorhexidine resistance among MRSA strains. Chlorhexidine resistance is associated with qac genes, which encode multidrug efflux pumps that increase bacterial tolerance to disinfectant agents. The global distribution and prevalence of qacA and qacB genes are highly variable. One study reported that qacA and qacB genes could be found in 0.9% - 83.3% of clinical MRSA isolates worldwide. The goal of this study was to determine the prevalence of chlorhexidine resistance and identify the resistance-associated genes from our MRSA samples using whole genome sequencing (WGS). Methods: 474 MRSA samples were obtained from hospitals in Detroit, MI (287) and Cleveland, OH (187). Whole genome sequencing was performed using the NextSeq (Illumina Inc., CA) platform. The sequencing data was analyzed using ResFinder 4.1, a publicly available database that can be used to identify acquired genes and chromosomal mutations mediating antimicrobial resistance. The output was organized into a data sheet to visualize the presence of the genes of interest. Results: The qacA gene was present in only one MRSA sample from the Cleveland area hospital. In the samples from Detroit, 14 out of 287 showed disinfectant resistance genes. The qacA, qacB, and qacD were present in 1, 6, and 7 samples, respectively. The prevalence of any qac gene in the Cleveland area samples was 0.5%. Meanwhile, the prevalence of any qac gene in Detroit area samples was 4.9%. Among the 7 samples that have qacD gene, 6 samples have more than one copy of qacD. Conclusions: The prevalence of the “qac” gene varied widely based on the origin of the samples. Detroit area samples had more qac genes prevalence than Cleveland area samples. Chlorhexidine is a widely used antiseptic/disinfectant, and it plays a vital role in reducing carriage and preventing infection among hospitalized patients colonized with MRSA. Monitoring and addressing MRSA-reduced susceptibility to chlorhexidine is imperative for maintaining the effectiveness of infection control practices such as decolonization.

Comparison of the effectiveness of core genome multilocus sequence typing and polymerase chain reaction-based open reading frame typing in tracing nosocomial methicillin-resistant Staphylococcus aureus transmission

IntroductionA clonal shift from staphylococcal cassette chromosome mec (SCCmec) type II/ST5 methicillin-resistant Staphylococcus aureus (MRSA) to SCCmec type IV/clonal complex (CC)1 MRSA has occurred rapidly in Japan. Our previous research in a geriatric hospital found SCCmec type IV/CC1 MRSA prevalence in long-term care wards. Due to intensive personal care requirements, frequent contact with healthcare providers can potentially cause unintentional nosocomial MRSA transmission. We performed polymerase chain reaction-based open reading frame typing (POT) and core genome multilocus sequence typing (cgMLST) to investigate the occurrence of nosocomial transmission and to compare the results of these methods. MethodsPOT and whole genome sequencing were performed in 83 MRSA isolates. Commercial automated software (Ridom SeqSphere+) was used to perform cgMLST. MRSA isolates with 0–8 allelic differences were considered related, and medical records were consulted in these cases. ResultsSCCmec type IV/CC1 MRSA was the most frequently detected clone (n = 56, 67.5 %), which was divided into 14 POT types, followed by SCCmec type I/ST8 (n = 9) and SCCmec type IV/ST8 (n = 8). Identical POT types were found across 7 of 11 wards. However, cgMLST analysis identified only three cases (six strains) of high genetic similarity, indicating nosocomial transmission; only one involved SCCmec type IV/CC1 (two strains). The mean allelic difference in the core genomes between strains with identical POT types in the same ward was 55.3 ± 22.0. ConclusionsThe cgMLST method proved more effective for identifying nosocomial transmissions compared to POT, highlighting its utility in tracking MRSA spread in healthcare settings.

Proteomic and metabolomic profiling of methicillin resistant versus methicillin sensitive Staphylococcus aureus using a simultaneous extraction protocol.

Understanding the biology of methicillin resistant Staphylococcus aureus (MRSA) is crucial to unlocking insights for new targets in our fight against this antimicrobial resistant priority pathogen. Although proteomics and metabolomic profiling offer the potential to elucidating such biological markers, reports of methodological approaches for carrying this out in S. aureus isolates remain limited. We describe the use of a dual-functionality methanol extraction method for the concurrent extraction of protein and metabolites from S. aureus and report on the comparative analysis of the proteomic and metabolomic profiles of MRSA versus methicillin sensitive S. aureus (MSSA). Bacterial reference strains MRSA ATCC43300 and MSSA ATCC25923 were used. The conventional urea methodology was used for protein extraction and a methanol based method was used for concurrent proteins and metabolites extraction. Proteomic and metabolomic profiling was carried out using TimsTOF mass spectrometry. Data processing was carried out using the MaxQuant version 2.1.4.0. This study represents the first report on the utilization of the methanol extraction method for concurrent protein and metabolite extraction in Gram positive bacteria. Our findings demonstrate good performance of the method for the dual extraction of proteins and metabolites from S. aureus with demonstration of reproducibility. Comparison of MRSA and MSSA strains revealed 407 proteins with significantly different expression levels. Enrichment analysis of those proteins revealed distinct pathways involved in fatty acid degradation, metabolism and beta-lactam resistance. Penicillin-binding protein PBP2a, the key determinant of MRSA resistance, exhibited distinct expression patterns in MRSA isolates. Metabolomic analysis identified 146 metabolites with only one exclusive to the MRSA. The enriched pathways identified were related to arginine metabolism and biosynthesis. Our findings demonstrate the effectiveness of the methanol-based dual-extraction method, providing simultaneous insights into the proteomic and metabolomic landscapes of S. aureus strains. These findings demonstrate the utility of proteomic and metabolomic profiling for elucidating the biological basis of antimicrobial resistance.

SCC mec typing among the clinical isolates of Methicillin Resistant Staphylococcus aureus (MRSA) from a Tertiary Care Hospital, Puducherry

Introduction: Methicillin resistance is exhibiteddue to presence of mecA gene and is commonly referred as SCCmec types in MRSA. SCCmec typing helps and acts a way to characterize MRSA infections and gain knowledge in its epidemiology. Our study aims to characterize mecA gene responsible in methicillin resistance by SCCmec typing and emphasize the use of SCCmec typing in routine diagnosis for the classification of MRSA. Methods: A total of 191 clinical isolates were included for the study.Documentation like clinical details and associated risk factors for MRSA acquisition were done. SCCmec typing (type I-V) was done by using M-PCR as per Boyeet al., 2007 protocol. Results: In our study, five different SCCmec types namely SCCmec type I, type II, type III, type IV and type V were detected among the clinical isolates of MRSA. Sccmec type V (53%) was found to be the most predominant type followed by SCCmec type III (47%) and SCCmec type IV (37.6%). Among the 191 MRSA isolates, 7 clinical isolates were SCCmec non-typeable isolates. Conclusion: Our study reports existence of multiple SCCmec types among the MRSA clinical isolates. SCCmec typing is helpful in determining the contribution of the isolates in understanding their association being CA-MRSA or HA-MRSA which in turn helps in designing effective surveillance and control strategies in the management of MRSA infections.

Molecular and Phylogenomic Analysis of a Vancomycin Intermediate Resistance USA300LV Strain in Chile.

Antimicrobial resistance is a major global health problem, and, among Gram-positive bacteria, methicillin-resistant Staphylococcus aureus (MRSA) represents a serious threat. MRSA causes a wide range of infections, including bacteremia, which, due to the limited use of β-lactams, is difficult to treat. This study aimed to analyze 51 MRSA isolates collected in 2018 from samples of patients with bacteremia from two hospitals of the Metropolitan Health Service of Santiago, Chile, both in their resistance profile and in the identification of virulence factors. In addition, genomic characterization was carried out by the WGS of an isolate that was shown to be the one of greatest concern (N°. 42) due to its intermediate resistance to vancomycin, multiple virulence factors and being classified as ST8 PVL-positive. In our study, most of the isolates turned out to be multidrug-resistant, but there are still therapeutic options, such as tetracycline, rifampicin, chloramphenicol and vancomycin, which are currently used for MRSA infections; however, 18% were PVL positive, which suggests greater virulence of these isolates. It was determined that isolate N°42 is grouped within the USA300-LV strains (ST8, PVL+, COMER+); however, it has been suggested that, in Chile, a complete displacement of the PVL-negative ST5 clone has not occurred.

Antibacterial and antibiofilm activities of zingerone and niosomal zingerone against methicillin-resistant Staphylococcus aureus (MRSA).

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of nosocomial and community acquired infections. Nanoparticles are considered as proper tools to overcome the therapeutic problem of antimicrobial-resistant infections because of the drug concentration increment at the desired location and protection from enzymatic degradation. The goal of this study was to evaluate the effect of the antibacterial and antibiofilm activities of zingerone and niosome containing zingerone against pre-formed biofilm of MRSA isolates. 62 MRSA isolates cultured from patients with diabetic ulcers were investigated. Niosomes were synthesized and characterized by X-ray diffraction, zeta potential and scanning electron microscopy (SEM). The size of niosomal particles measured by SEM and zetasizer. The surface charge of prepared niosomes was about -37 mV. The effect of the zingerone and noisome containing zingerone was evaluated against biofilms of MRSA isolates. Also, the antibiofilm activity of prepared niosomes on gene expression of MRSA biofilms was evaluated using Real Time PCR. Our results demonstrated that the niosome containing zingerone had a diameter of 196.1 nm and a -37.3-mV zeta potential. Zingerone removed one and three-day old biofilms of MRSA at the concentration of 1000 μg/ml, while the zingerone-laoded niosomes removed 1, 3- and 5-days old biofilms at the concentration of 250 μg/ml, 250 μg/ml, and 500 μg/ml. The results indicated that niosome containing zingerone eliminated MRSA and its biofilms faster compared with free zingerone and it suggested that zingerone-encapsulated niosomes could be considered as a promising treatment against MRSA and its biofilms.

Phenotypic and Molecular Characterization of Vancomycin and Linezolid Resistance in Clinical Isolates of Staphylococcus aureus

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of healthcare-associated infections and poses a significant global challenge due to its high pathogenicity and rapid spread among patients. Vancomycin and linezolid are key antibiotics for treating staphylococcal MRSA infections. Objectives: This study aimed to investigate resistance to vancomycin and linezolid in clinical isolates of S. aureus. Methods: In this study, 270 S. aureus isolates were collected from patients admitted to hospitals in Tehran and Qazvin. Strain identification was performed using biochemical methods and femA gene amplification. Resistance to methicillin and vancomycin was determined using disk diffusion, minimum inhibitory concentration (MIC) determination, and E-test methods. Methicillin resistance was also assessed by detecting the presence of the mecA and mecC genes. Resistance to vancomycin and linezolid was evaluated by examining the presence of vanA, B, C, and cfr genes using PCR and sequencing. Results: Out of 270 S. aureus isolates, 152 (56.3%) were identified as MRSA, with 144 (94.7%) of these MRSA isolates containing the mecA gene. Two (0.7%) VRSA isolates were observed, and the vanA gene was detected in both. Additionally, four VISA (1.4%) isolates were identified, but the vanA gene was not detected in any of the VISA strains. None of the isolates contained the mecC, vanB, or vanC genes. Among the 10 (3.7%) linezolid-resistant isolates, none contained the cfr gene. Conclusions: The results of this study demonstrated a high prevalence of MRSA isolates, which can lead to treatment failure with beta-lactam antibiotics. The emergence and spread of VRSA and VISA isolates pose a serious concern for the healthcare system, particularly in the treatment of MRSA infections. Therefore, the findings emphasize the need for appropriate monitoring and control measures to prevent the emergence and transmission of MRSA and VRSA strains. Vancomycin and linezolid remain suitable drugs for treating these infections.

Genomic investigation and clinical correlates of the in vitro β-lactam: NaHCO3 responsiveness phenotype among methicillin-resistant Staphylococcus aureus isolates from a randomized clinical trial.

NaHCO3 responsiveness is a novel phenotype where some methicillin-resistant Staphylococcus aureus (MRSA) isolates exhibit significantly lower minimal inhibitory concentrations (MIC) to oxacillin and/or cefazolin in the presence of NaHCO3. NaHCO3 responsiveness correlated with treatment response to β-lactams in an endocarditis animal model. We investigated whether treatment of NaHCO3-responsive strains with β-lactams was associated with faster clearance of bacteremia. The CAMERA2 trial (Combination Antibiotics for Methicillin-Resistant Staphylococcus aureus) randomly assigned participants with MRSA bloodstream infections to standard therapy, or to standard therapy plus an anti-staphylococcal β-lactam (combination therapy). For 117 CAMERA2 MRSA isolates, we determined by broth microdilution the MIC of cefazolin and oxacillin, with and without 44 mM of NaHCO3. Isolates exhibiting ≥4-fold decrease in the MIC to cefazolin or oxacillin in the presence of NaHCO3 were considered "NaHCO3-responsive" to that agent. We compared the rate of persistent bacteremia among participants who had infections caused by NaHCO3-responsive and non-responsive strains, and that were assigned to combination treatment with a β-lactam. Thirty-one percent (36/117) and 25% (21/85) of MRSA isolates were NaHCO3-responsive to cefazolin and oxacillin, respectively. The NaHCO3-responsive phenotype was significantly associated with sequence type 93, SCCmec type IVa, and mecA alleles with substitutions in positions -7 and -38 in the regulatory region. Among participants treated with a β-lactam, there was no association between the NaHCO3-responsive phenotype and persistent bacteremia (cefazolin, P = 0.82; oxacillin, P = 0.81). In patients from a randomized clinical trial with MRSA bloodstream infection, isolates with an in vitro β-lactam-NaHCO3-responsive phenotype were associated with distinctive genetic signatures, but not with a shorter duration of bacteremia among those treated with a β-lactam.

Antimicrobial Susceptibility of Various MRSA Clinical Isolates and the Impact of Glycopeptide MICs on Clinical and Microbiological Outcomes.

While vancomycin has remained the mainstay of the treatment for methicillin-resistant Staphylococcus aureus (MRSA) infections, there is growing evidence of the clinical impact of increased glycopeptide minimum inhibitory concentrations (MICs) in MRSA isolates. This study aimed to determine the susceptibility of various MRSA isolates to different antibiotics with antistaphylococcal activity and the impact of glycopeptide MICs on clinical and microbiological outcomes. This retrospective cohort study, conducted between 2013 and 2017, evaluated the susceptibility of MRSA strains isolated from various clinical samples to antistaphylococcal antibiotics using the gradient strip method. The clinical and laboratory features of patients infected with MRSA isolates with elevated glycopeptide MICs (>1 mg/L) and with isolates that had low glycopeptide MICs (≤1 mg/L) were compared. A total of 104 patients infected with MRSA strains were included in this study. Male sex (odds ratio [OR]=2.48, 95% confidence interval [CI]=1.01-6.10, p=0.048), two or more comorbidities (OR=2.48, 95% CI=1.03-6.50, p=0.044), history of MRSA infection (OR=4.91, 95% CI=1.70-14.28, p=0.003) and a longer hospital stay prior to MRSA infection (OR=2.32, 95% CI=1.05-7.85, p=0.040) were independent risk factors for high glycopeptide MICs. In MRSA infections with a teicoplanin MIC of >0.75mg/L, the microbiological and treatment failures were 46.2% (p=0.044) and 60.6% (p=0.042), respectively. This study showed that the critical MIC value, which suggested treatment failure as well as microbiological failure in the teicoplanin-treated MRSA infections, was >0.75 mg/L rather than >1 mg/L in our study cohort. The identification of high-risk patients;for treatment failures and mortality considering gradient strip method MIC values is crucial for the effective management of MRSA infections.