Methicillin-resistant S. Aureus Infections Research Articles

Genomic characterization of Staphylococcus aureus isolated from patients admitted to intensive care units of a tertiary care hospital: epidemiological risk of nasal carriage of virulent clone during admission.

We conducted a molecular epidemiological study of Staphylococcus aureus using whole-genome sequence data and clinical data of isolates from nasal swabs of patients admitted to the intensive care unit (ICU) of Hiroshima University hospital. The relationship between isolate genotypes and virulence factors, particularly for isolates that caused infectious diseases during ICU admission was compared with those that did not. The nasal carriage rates of methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) in patients admitted to the ICU were 7.0% and 20.1%, respectively. The carriage rate of community-acquired (CA)-MRSA was 2.3%, accounting for 32.8% of all MRSA isolates. Whole-genome sequencing analysis of the MRSA isolates indicated that most, including CA-MRSA and healthcare-associated (HA)-MRSA, belonged to clonal complex (CC) 8 [sequence type (ST) 8] and SCCmec type IV. Furthermore, results for three disease foci (pneumonia, skin and soft tissue infection, and deep abscess) and the assessment of virulence factor genes associated with disease conditions [bacteremia, acute respiratory distress syndrome (ARDS), disseminated intravascular coagulopathy (DIC), and septic shock] suggested that nasal colonization of S. aureus clones could represent a risk for patients within the ICU. Particularly, MRSA/J and MSSA/J may be more likely to cause deep abscess infection; ST764 may cause ventilation-associated pneumonia, hospital-acquired pneumonia and subsequent bacteremia, and ARDS, and tst-1-positive isolates may cause DIC onset.IMPORTANCENasal colonization of MRSA in patients admitted to the intensive care unit (ICU) may predict the development of MRSA infections. However, no bacteriological data are available to perform risk assessments for Staphylococcus aureus infection onset. In this single-center 2-year genomic surveillance study, we analyzed all S. aureus isolates from nasal swabs of patients admitted to the ICU and those from the blood or lesions of in-patients who developed infectious diseases in the ICU. Furthermore, we identified the virulent clones responsible for causing infectious diseases in the ICU. Herein, we report several virulent clones present in the nares that are predictive of invasive infections. This information may facilitate the design of preemptive strategies to identify and eradicate virulent MRSA strains, reducing nosocomial infections within the ICU.

Collaborative Cross mice have diverse phenotypic responses to infection with Methicillin-resistant Staphylococcus aureus USA300.

Staphylococcus aureus (S. aureus) is an opportunistic pathogen causing diseases ranging from mild skin infections to life threatening conditions, including endocarditis, pneumonia, and sepsis. To identify host genes modulating this host-pathogen interaction, we infected 25 Collaborative Cross (CC) mouse strains with methicillin-resistant S. aureus (MRSA) and monitored disease progression for seven days using a surgically implanted telemetry system. CC strains varied widely in their response to intravenous MRSA infection. We identified eight 'susceptible' CC strains with high bacterial load, tissue damage, and reduced survival. Among the surviving strains, six with minimal colonization were classified as 'resistant', while the remaining six tolerated higher organ colonization ('tolerant'). The kidney was the most heavily colonized organ, but liver, spleen and lung colonization were better correlated with reduced survival. Resistant strains had higher pre-infection circulating neutrophils and lower post-infection tissue damage compared to susceptible and tolerant strains. We identified four CC strains with sexual dimorphism: all females survived the study period while all males met our euthanasia criteria earlier. In these CC strains, males had more baseline circulating monocytes and red blood cells. We identified several CC strains that may be useful as new models for endocarditis, myocarditis, pneumonia, and resistance to MRSA infection. Quantitative Trait Locus (QTL) analysis identified two significant loci, on Chromosomes 18 and 3, involved in early susceptibility and late survival after infection. We prioritized Npc1 and Ifi44l genes as the strongest candidates influencing survival using variant analysis and mRNA expression data from kidneys within these intervals.

Physalin H, physalin B, and isophysalin B suppress the quorum-sensing function of Staphylococcus aureus by binding to AgrA.

The virulence of Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), depends on the expression of toxins and virulence factors controlled by the quorum-sensing (QS) system, encoded on the virulence accessory gene regulator (agr) locus. The aim of this study was to identify a phytochemical that inhibits Agr-QS function and to elucidate its mechanism. We screened 577 compounds and identified physalin H, physalin B, and isophysalin B--phytochemicals belonging to physalins found in plants of the Solanaceae family--as novel Agr-QS modulators. Biological analyses and in vitro protein-DNA binding assays suggested that these physalins suppress gene expression related to the Agr-QS system by inhibiting binding of the key response regulator AgrA to the agr promoters, reducing the function of hemolytic toxins downstream of these genes in MRSA. Furthermore, although physalin F suppressed gene expression in the Agr-QS system, its anti-hemolytic activity was lower than that of physalins H, B, and isophysalin B. Conversely, five physalins isolated from the same plant with the ability to suppress Agr-QS did not reduce bacterial Agr-QS activity but inhibited AgrA binding to DNA in vitro. A docking simulation revealed that physalin interacts with the DNA-binding site of AgrA in three docking states. The carbonyl oxygens at C-1 and C-18 of physalins, which can suppress Agr-QS, were directed to residues N201 and R198 of AgrA, respectively, whereas these carbonyl oxygens of physalins, without Agr-QS suppression activity, were oriented in different directions. Next, 100-ns molecular dynamics simulations revealed that the hydrogen bond formed between the carbonyl oxygen at C-15 of physalins and L186 of AgrA functions as an anchor, sustaining the interaction between the carbonyl oxygen at C-1 of physalins and N201 of AgrA. Thus, these results suggest that physalin H, physalin B, and isophysalin B inhibit the interaction of AgrA with the agr promoters by binding to the DNA-binding site of AgrA, suppressing the Agr-QS function of S. aureus. Physalins that suppress the Agr-QS function are proposed as potential lead compounds in the anti-virulence strategy for MRSA infections.

Dry and liquid formulations of IBT-V02, a novel multi-component toxoid vaccine, are effective against Staphylococcus aureus isolates from low-to-middle income countries.

Staphylococcus aureus is the leading cause of skin and soft tissue infections (SSTIs) in the U.S. as well as more serious invasive diseases, including bacteremia, sepsis, endocarditis, surgical site infections, osteomyelitis, and pneumonia. These infections are exacerbated by the emergence of antibiotic-resistant clinical isolates such as methicillin-resistant S. aureus (MRSA), highlighting the need for alternatives to antibiotics to treat bacterial infections. We have previously developed a multi-component toxoid vaccine (IBT-V02) in a liquid formulation with efficacy against multiple strains of Staphylococcus aureus prevalent in the industrialized world. However, liquid vaccine formulations are not compatible with the paucity of cold chain storage infrastructure in many low-to-middle income countries (LMICs). Furthermore, whether our IBT-V02 vaccine formulations are protective against S. aureus isolates from LMICs is unknown. To overcome these limitations, we developed lyophilized and spray freeze-dried formulations of IBT-V02 vaccine and demonstrated that both formulations had comparable biophysical attributes as the liquid formulation, including similar levels of toxin neutralizing antibodies and protective efficacy against MRSA infections in murine and rabbit models. To enhance the relevancy of our findings, we then performed a multi-dimensional screen of 83 S. aureus clinical isolates from LMICs (e.g., Democratic Republic of Congo, Palestine, and Cambodia) to rationally down-select strains to test in our in vivo models based on broad expression of IBT-V02 targets (i.e., pore-forming toxins and superantigens). IBT-V02 polyclonal antisera effectively neutralized toxins produced by the S. aureus clinical isolates from LMICs. Notably, the lyophilized IBT-V02 formulation exhibited significant in vivo efficacy in various preclinical infection models against the S. aureus clinical isolates from LMICs, which was comparable to our liquid formulation. Collectively, our findings suggested that lyophilization is an effective alternative to liquid vaccine formulations of our IBT-V02 vaccine against S. aureus infections, which has important implications for protection from S. aureus isolates from LMICs.

Exploring secretory proteome and cytokine kinetic of human peripheral blood mononuclear cells exposed to methicillin-resistant Staphylococcus aureus biofilms and planktonic bacteria.

Staphylococcus aureus is a highly successful pathogen infecting various body parts and forming biofilms on natural and artificial surfaces resulting in difficult-to-treat and chronic infections. We investigated the secreted cytokines and proteomes of isolated peripheral blood mononuclear cells (PBMCs) from healthy volunteers exposed to methicillin-resistant S. aureus (MRSA) biofilms or planktonic bacteria. Additionally, the cytokine profiles in sera from patients with community-acquired pneumonia (CAP) caused by S. aureus were investigated. The aim was to gain insights into the immune response involved and differentiate between the planktonic and sessile MRSA forms. We identified 321 and 298 targets that were significantly differently expressed in PBMCs when exposed to planktonic or biofilm-embedded bacteria, respectively. PBMCs exposed to planktonic MRSA cells secreted increased levels of TNF-α, while IL-18 was elevated when exposed to the biofilm. The machine-learning analyses of the cytokine profiles obtained for the in vitro PBMCs and CAP sera distinguished between the two types of bacteria forms based on cytokines IL-18, IL12, and IL-17, and with a lower importance IL-6. Particularly, IL-18 which has not been correlated with S. aureus biofilms so far might represent a suitable marker for monitoring chronification during MRSA infection to individualize the therapy, but this hypothesis must be proved in clinical trials.

Characterization of the broad-spectrum antibacterial activity of bacteriocin-like inhibitory substance-producing probiotics isolated from fermented foods

Antimicrobial peptides, such as bacteriocin, produced by probiotics have become a promising novel class of therapeutic agents for treating infectious diseases. Selected lactic acid bacteria (LAB) isolated from fermented foods with probiotic potential were evaluated for various tests, including exopolysaccharide production, antibiotic susceptibility, acid and bile tolerance, antibacterial activity, and cell adhesion and cytotoxicity to gastric cell lines. Six selected LAB strains maintained their high viability under gastrointestinal conditions, produced high exopolysaccharides, showed no or less cytotoxicity, and adhered successfully to gastric cells. Furthermore, three strains, Weissella confusa CYLB30, Lactiplantibacillus plantarum CYLB47, and Limosilactobacillus fermentum CYLB55, demonstrated a strong antibacterial effect against drug-resistant Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica serovar Choleraesuis, Enterococcus faecium, and Staphylococcus aureus. Whole genome sequencing was performed on these three strains using the Nanopore platform; then, the results showed that all three strains did not harbor genes related to toxins, superantigens, and acquired antimicrobial resistance, in their genome. The bacteriocin gene cluster was found in CYLB47 genome, but not in CYLB30 and CYLB55 genomes. In SDS-PAGE, the extract of CYLB30 and CYLB47 bacteriocin-like inhibitory substance (BLIS) yielded a single band with a size of less than 10 kDa. These BLIS inhibited the growth and biofilm formation of drug-resistant P. aeruginosa and methicillin-resistant S. aureus (MRSA), causing membrane disruption and inhibiting adhesion ability to human skin HaCaT cells. Moreover, CYLB30 and CYLB47 BLIS rescued the larvae after being infected with P. aeruginosa and MRSA infections. In conclusion, CYLB30 and CYLB47 BLIS may be potential alternative treatment for multidrug-resistant bacteria infections.

A novel extraction-free dual HiFi-LAMP assay for detection of methicillin-sensitive and methicillin-resistant Staphylococcus aureus.

Staphylococcus aureus (S. aureus) is a leading cause of bacteremia and blood stream infections. Methicillin-resistant S. aureus (MRSA) that first appeared in 1961 often caused hospital-acquired infections (HAIs) and community-acquired infections (CAIs) and was associated with high mortality rate. Accurate and rapid point-of-care testing (POCT) of MRSA is crucial for clinical management and treatment of MRSA infections, as well as the prevention and control of HAIs and CAIs. Here, we reported a novel extraction-free dual HiFi-LAMP assay for discriminative detection of methicillin-susceptible S. aureus and MRSA. The dual HiFi-LAMP assay can detect 30 copies/reaction of nuc and mecA genes with detection limits of 147 and 158 copies per 25 µL reaction, respectively. A retrospective clinical evaluation with 107 clinical S. aureus isolates showed both sensitivity and specificity of 100%. A prospective clinical evaluation with 35 clinical samples revealed a specificity of 100% and a sensitivity of 92.3%. The dual HiFi-LAMP assay can detect almost all S. aureus samples (141/142; 99.3%) within 20 min, implying that the entire HiFi-LAMP assay (including sample process) can be completed within 40 min, extremely significantly shorter than 3-5 days by the traditional clinical microbial culture and antibiotic susceptibility testing. The novel extraction-free dual HiFi-LAMP assay can be used as a robust POCT tool to promote precise diagnosis and treatment of MRSA infections in hospitals and to facilitate surveillance of MRSA at hospital and community settings.IMPORTANCEMethicillin-resistant Staphylococcus aureus (MRSA) was associated with high mortality rate and listed as a "priority pathogen" by the World Health Organization. Accurate and rapid point-of-care testing (POCT) of MRSA is critically required for clinical management and treatment of MRSA infections. Some previous LAMP-based POCT assays for MRSA might be questionable due to their low specificity and the lack of appropriate evaluation directly using clinical samples. Furthermore, they are relatively tedious and time-consuming because they require DNA extraction and lack multiplex detection capacity. Here, we reported a novel extraction-free dual HiFi-LAMP assay for discriminative detection of MRSA and methicillin-susceptible S. aureus. The assay has high specificity and sensitivity and can be completed within 40 min. Clinical evaluation with real clinical samples and clinical isolates showed excellent performance with 100% specificity and 92.3%-100% sensitivity. The novel extraction-free assay may be a robust POCT tool to promote precise diagnosis of MRSA infections and facilitate surveillance of MRSA at hospital and community settings.

The active ingredients in Chinese peony pods synergize with antibiotics to inhibit MRSA growth and biofilm formation

Staphylococcus aureus (S. aureus) is a zoonotic pathogen that infects both humans and animals. The rapid spread of methicillin-resistant S. aureus (MRSA) and its resistance to antibiotics, along with its ability to form biofilms, poses a serious challenge to the clinical application of traditional antibiotics. Peony (Paeonia lactiflora Pall.) is a traditional Chinese medicine with multiple pharmacological effects. This study observed the strong antibacterial and antibiofilm activity of the water extract (WE) and ethyl acetate extract (EA) of Chinese peony pods against MRSA. The combination of EA and vancomycin, cefotaxime, penicillin G or methicillin showed a synergistic or additive antibacterial and antibiofilm effects on MRSA, which is closely related to the interaction of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (PG) and methyl gallate (MG). The active ingredients in peony pods have been found to increase the sensitivity of MRSA to antibiotics and demonstrate antibiofilm activity, which is mainly related to the down-regulation of global regulatory factors sarA and sigB, extracellular PIA and eDNA encoding genes icaA and cdiA, quorum sensing related genes agrA, luxS, rnaIII, hld, biofilm virulence genes psma and sspA, and genes encoding clotting factors coa and vwb, but is not related to genes that inhibit cell wall anchoring. In vivo test showed that both WE and EA were non-toxic and significantly prolonged the lifespan of G. mellonella larvae infected with MRSA. This study provides a theoretical basis for further exploration of the combined use of PG, MG and antibiotics to combat MRSA infections.

Oral and rectal colonization of methicillin-resistant Staphylococcus aureus in long-term care facility residents and their association with clinical status.

Staphylococcus aureus is a commensal bacterium in humans, but it sometimes causes opportunistic infectious diseases such as suppurative skin disease, pneumonia, and enteritis. Therefore, it is important to determine the prevalence of S. aureus and methicillin-resistant S. aureus (MRSA) in individuals, especially older adults. In this study, we investigated the prevalence of S. aureus and MRSA in the oral cavity and feces of residents in long-term care facilities (LTCFs). S. aureus was isolated from the oral cavity of 61/178 (34.3%) participants, including 28 MRSA-positive participants (15.7%), and from the feces of 35/127 (27.6%) participants, including 16 MRSA-positive participants (12.6%). S. aureus and MRSA were isolated from both sites in 19/127 individuals (15.0%) and 10/127 individuals (7.9%), respectively. Among 19 participants with S. aureus isolation from both sites, 17 participants showed the same sequence type (ST) type. Then, we analyzed the correlation of S. aureus and MRSA in the oral cavity and rectum with the participant's condition. S. aureus and MRSA positivity in the oral cavity was significantly related to tube feeding, while there was no correlation of rectal S. aureus/MRSA with any factors. Our findings regarding the oral inhabitation of MRSA and its risk factors indicate the importance of considering countermeasures against MRSA infection in LTCFs.

Effect of Bioactive Peptide, KR-12-a5, on Growth of Clinical Methicillin-resistant Staphylococcus aureus Isolates

Staphylococcus aureus is one of the most frequent infectious agents causing hospitals and community associated infections. Developing the treatment for methicillin resistant S. aureus (MRSA) is a challenging arena for effective therapeutics. Application of antimicrobial peptides is an alternative option to control MRSA infection. The potential of the bioactive peptide against MRSA was previously revealed; however, the study on the effect of KR-12-a5 on the various clinical MRSA growth was not elucidated. This study aimed to investigate the effect of bioactive peptide KR-12-a5 on the growth of MRSA isolated from patients. Evaluation of KR-12-a5 peptide using antimicrobial susceptibility test was performed. Antimicrobial activity assay showed that the KR-12-a5 at 2-64 μM could inhibit the growth of S. aureus ATCC 29213 in the range of 55.0±0.8%-81.0±6.2%. In case of KR-12-a5 treatment at 2, 4, and 8 μM, growth of all twenty clinical MRSA isolates could be inhibited less than 90% in the range of 0.0-75.9±1.6%, 0.0-73.0±4.2%, and 0.0-77.1±1.7%, respectively. Whereas the treatment of those at 16, 32, and 64 μM, growth of clinical MRSA could be significantly inhibited more than 90% (p < 0.05) in the range of 94.3±6.7%-100.4±0.4% in 3 isolates (15%), 97.3±0.3%-99.5±0.2% in 4 isolates (20%), and 91.6±0.8%-99.0±0.1% in 9 isolates (45%), respectively. These findings indicated that the KR-12-a5 effectively controls growth of both S. aureus ATCC 29213 and clinical MRSA isolates

Assessing Tetracycline susceptibility in multidrug-resistant Staphylococcus aureus: A clinical study in Kathmandu

Introduction: Staphylococcus aureus, recognized as an opportunistic pathogen, is frequently implicated in diverse human infections. Managing multidrug-resistant S. aureus poses a significant challenge in the clinical sector. Beyond B-lactam antibiotics, healthcare practitioners commonly prescribe Glycopeptides, Tetracycline, Quinolones, and Macrolide-Lincosamide-Streptogramin B antibiotics as alternative treatments for such infections. Consequently, this study was undertaken to evaluate the susceptibility of multidrug-resistant S. aureus to tetracycline in clinical samples. Methods: The study spanned 15 months, from March 2020 to May 2021, during which clinical samples were collected from three distinct hospitals in Kathmandu-Dirghayu Guru Hospital, Chabahil; Bhaktapur Cancer Hospital, Bhaktapur; and Shankarapur Hospital, Jorpati. Sample processing and laboratory work were conducted in the Microbiology Laboratory of Dirghyau Guru Hospital to assess the prevalence of different multidrug-resistant (MDR) phenotypes among S. aureus and their correlation with Methicillin resistance. The study included 266 isolates of S. aureus, and Methicillin resistance was determined using the Cefoxitin disc diffusion method. Results: Among 2004 clinical samples processed, 266 (13.27%) isolates were S. aureus. The multidrug resistant strain isolated from Outpatient department was 56.84% (54/95) and from inpatient department, the data was 43.15% (41/95). Among the 266 S. aureus, 28.94% Methicillin resistant S. aureus (MRSA) and 71.05% Methicillin sensitive S. aureus (MSSA) isolates were detected. Tetracycline resistance was found in 9.8% of total S. aureus isolates whereas 27.36% (26/95) among multidrug resistant S. aureus isolates. Resistance to Tetracycline was higher in MRSA as compared to MSSA. Conclusion: The detection rate of MRSA emphasizes the urgency to improve healthcare practices to control the spread of MRSA infections. Tetracycline can be used as a viable alternative antibiotic for combating S. aureus infections, particularly in Kathmandu.

Antibiofilm and Antivirulence Properties of 6-Polyaminosteroid Derivatives against Antibiotic-Resistant Bacteria.

The emergence of multi-drug resistant pathogens is a major public health problem, leading us to rethink and innovate our bacterial control strategies. Here, we explore the antibiofilm and antivirulence activities of nineteen 6-polyaminosterol derivatives (squalamine-based), presenting a modulation of their polyamine side chain on four major pathogens, i.e., carbapenem-resistant A. baumannii (CRAB) and P. aeruginosa (CRPA), methicillin-resistant S. aureus (MRSA), and vancomycin-resistant E. faecium (VRE) strains. We screened the effect of these derivatives on biofilm formation and eradication. Derivatives 4e (for CRAB, VRE, and MRSA) and 4f (for all the strains) were the most potent ones and displayed activities as good as those of conventional antibiotics. We also identified 11 compounds able to decrease by more than 40% the production of pyocyanin, a major virulence factor of P. aeruginosa. We demonstrated that 4f treatment acts against bacterial infections in Galleria mellonella and significantly prolonged larvae survival (from 50% to 80%) after 24 h of CRAB, VRE, and MRSA infections. As shown by proteomic studies, 4f triggered distinct cellular responses depending on the bacterial species but essentially linked to cell envelope. Its interesting antibiofilm and antivirulence properties make it a promising a candidate for use in therapeutics.

1656. Evaluation of Using Oxacillin versus Vancomycin for Empiric Therapy of Late-Onset Neonatal Sepsis in NICU

Abstract Background Our Pediatric Antimicrobial Stewardship Program (PedASP) assessed methicillin-sensitive (MSSA) and methicillin-resistant S. aureus (MRSA) infections in neonatal intensive care unit (NICU) and created a NICU-specific antibiogram. Due to a rising trend of NICU MSSA infections seen in our 84-bed NICU, empiric therapy of late-onset neonatal sepsis (LONS) was modified from vancomycin to oxacillin in January 2021. We report the two-year results of using empiric oxacillin for LONS.Figure 1.Summary of S. aureus identified in LONS cases Methods We assessed MSSA versus MRSA isolated from blood and non-blood cultures in NICU between January 2019 and December 2022. Infants with weight less than 750 grams, previous MRSA infections, maternal MRSA colonization, or serious infections based on clinical judgement were advised to use empiric vancomycin. We examined the vancomycin eligibility criteria, antibiotic-pathogen mismatch rate, 30-day mortality, and frequency of peripheral intravenous (PIV) access loss after vancomycin or oxacillin administration. The cost-savings was estimated by comparing drug and lab costs associated with vancomycin and oxacillin use. Results LONS cases are summarized in Figure 1. There was no statistical difference in incidence of S. aureus bacteremia before and after protocol implementation (p=0.45). Antibiotic-pathogen mismatch rate was 0%. Cases of MRSA bacteremia increased from 0 to 3 post protocol initiation; all cases met vancomycin eligibility criteria. The 30-day mortality among MRSA infections increased from 0 to 1 case. Post protocol implementation, the PIV loss rate decreased from 4.7 to 3.3% (p=0.04), and vancomycin use decreased by 1154 doses while oxacillin use increased by 485 doses. Based on antibiotic use and 141 vancomycin levels averted, the estimated cost-savings was $140,229. Conclusion Our PedASP utilized local susceptibility data to change empiric antibiotics for LONS, reinforcing the benefit of using antibiogram to optimize antibiotic use in NICU. Implementing a NICU-specific antibiogram improved the effectiveness of antibiotic treatment and achieved significant cost savings. Disclosures All Authors: No reported disclosures

Studying the Anti-Virulence Activity of Meta-Bromo-Thiolactone against Staphylococcus aureus and MRSA Phenotypes

Quorum-sensing inhibitors have recently garnered great interest, as they reduce bacterial virulence, lower the probability of resistance, and inhibit infections. In this work, meta-bromo-thiolactone (mBTL), a potent quorum and virulence inhibitor of Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA), was formulated in chitosan nanoparticles (ChNPs) using the ionic gelation method. The mBTL-loaded-ChNPs were characterized by their particle size, polydispersity index, zeta potential, morphology, and drug release profile. The results show that the mBTL-loaded-CNPs comprised homogenized, spherical nanoparticles ranging from 158 ± 1.3 to 284 ± 5.6 nm with a sustainable release profile over 48 h at 37 °C. These findings confirm the successful preparation of mBTL-loaded-ChNPs. Confocal laser scanning microscopy showed a significant reduction in the number of viable cells, indicating the antibacterial efficacy of mBTL. Biofilms were observed by scanning electron microscopy, which showed that the bacterial cells in the control experiment were enclosed in thick biofilms. In the presence of mBTL, the bacterial cells remained disordered and did not form a biofilm. mBTL-loaded-ChNPs represent a potential approach to overcoming antimicrobial resistance in the treatment of MRSA infection.

Machine learning to predict risk for community-onset Staphylococcus aureus infections in children living in southeastern United States.

Staphylococcus aureus (S. aureus) is known to cause human infections and since the late 1990s, community-onset antibiotic resistant infections (methicillin resistant S. aureus (MRSA)) continue to cause significant infections in the United States. Skin and soft tissue infections (SSTIs) still account for the majority of these in the outpatient setting. Machine learning can predict the location-based risks for community-level S. aureus infections. Multi-year (2002-2016) electronic health records of children <19 years old with S. aureus infections were queried for patient level data for demographic, clinical, and laboratory information. Area level data (Block group) was abstracted from U.S. Census data. A machine learning ecological niche model, maximum entropy (MaxEnt), was applied to assess model performance of specific place-based factors (determined a priori) associated with S. aureus infections; analyses were structured to compare methicillin resistant (MRSA) against methicillin sensitive S. aureus (MSSA) infections. Differences in rates of MRSA and MSSA infections were determined by comparing those which occurred in the early phase (2002-2005) and those in the later phase (2006-2016). Multi-level modeling was applied to identify risks factors for S. aureus infections. Among 16,124 unique patients with community-onset MRSA and MSSA, majority occurred in the most densely populated neighborhoods of Atlanta's metropolitan area. MaxEnt model performance showed the training AUC ranged from 0.771 to 0.824, while the testing AUC ranged from 0.769 to 0.839. Population density was the area variable which contributed the most in predicting S. aureus disease (stratified by CO-MRSA and CO-MSSA) across early and late periods. Race contributed more to CO-MRSA prediction models during the early and late periods than for CO-MSSA. Machine learning accurately predicts which densely populated areas are at highest and lowest risk for community-onset S. aureus infections over a 14-year time span.

Knowledge, Practices, and Nasal Carriage Rate of MRSA Amongst Dental Professionals

Resistance of Staphylococcus aureus (S. aureus) against common antibiotics has been increasing given the rampant use of antibiotics. Methicillin-resistant S. aureus (MRSA) is particularly important because it has been reported to present in hospital-acquired as well as community-acquired infections. Our aim was to study the types and subtypes of MRSA isolated from nasal swabs taken from volunteering dental school students and to assess and analyse the knowledge of dental health care workers (DHCWs) on MRSA and prevention of its infection. A total of 100 participants, which included staff, students, and dental auxiliaries, from the School of Dental Sciences, Krishna Institute of Medical Sciences, Deemed to Be University, Karad, India, were included in the study which was conducted from June 2020 to 1 June 2021. All the participants completed a survey and underwent collection of nasal swabs. The samples were cultured and underwent microbiological and molecular analysis for MRSA. The questionnaire was sent out electronically via SurveyMonkey. Sixty-three percent of the participants were female. The majority were undergraduate students (66%) and younger than 25 years (77%). The prevalence of MRSA in the sample was the MRSA type SCCmec type V (54.8%). The survey reports a lack of knowledge amongst dental students on MRSA and prevention of its infection. The majority (69%) of participants had not heard of MRSA infection and had inadequate knowledge of MRSA infection. Only 29% of the participants think that a healthy person may have MRSA without feeling ill. Eighty-five percent of the participants reported that they have written polices and guidelines to control MRSA. Eighty-five percent of the participants indicated that they are less confident in their knowledge on MRSA infection control protocols, and 94% mentioned that the dental clinics have written guidelines for hand hygiene. DHCWs have limited knowledge of MRSA infection, emphasising the need for appropriate clinical training. Academic institutions should promote effective infection control training to protect students, faculty, and other employees.

Harnessing diversity and antagonism within the pig skin microbiota to identify novel mediators of colonization resistance to methicillin-resistant Staphylococcus aureus.

The microbiota mediate multiple aspects of skin barrier function, including colonization resistance to pathogens such as Staphylococcus aureus. The endogenous skin microbiota limits S. aureus colonization via competition and direct inhibition. Novel mechanisms of colonization resistance are promising therapeutic targets for drug-resistant infections, such as those caused by methicillin-resistant S. aureus (MRSA). Here, we developed and characterized a swine model of topical microbiome perturbation and MRSA colonization. As in other model systems, topical antimicrobial treatment had a little discernable effect on community diversity though the overall microbial load was sensitive to multiple types of intervention, including swabbing. In parallel, we established a porcine skin culture collection and screened 7,700 isolates for MRSA inhibition. Using genomic and phenotypic criteria, we curated three isolates to investigate whether prophylactic colonization would inhibit MRSA colonization in vivo. The three-member consortium together, but not individually, provided protection against MRSA colonization, suggesting cooperation and/or synergy among the strains. Inhibitory isolates were represented across all major phyla of the pig skin microbiota and did not have a strong preference for inhibiting closely related species, suggesting that relatedness is not a condition of antagonism. These findings reveal the porcine skin as an underexplored reservoir of skin commensal species with the potential to prevent MRSA colonization and infection. IMPORTANCE The skin microbiota is protective against pathogens or opportunists such as S. aureus, the most common cause of skin and soft tissue infections. S. aureus can colonize normal skin and nasal passages, and colonization is a risk factor for infection, especially on breach of the skin barrier. Here, we established a pig model to study the competitive mechanisms of the skin microbiota and their role in preventing colonization by MRSA. This drug-resistant strain is also a livestock pathogen, and swine herds can be reservoirs of MRSA carriage. From 7,700 cultured skin isolates, we identified 37 unique species across three phyla that inhibited MRSA. A synthetic community of three inhibitory isolates provided protection together, but not individually, in vivo in a murine model of MRSA colonization. These findings suggest that antagonism is widespread in the pig skin microbiota, and these competitive interactions may be exploited to prevent MRSA colonization.

The clinical spectrum of Staphylococcus aureus infections in children admitted to Chris Hani Baragwanath Academic Hospital, South Africa: A retrospective, descriptive study

Background. Staphylococcus aureus infection is associated with considerable morbidity and mortality. There are relatively few studiesdescribing invasive S. aureus infections in children, particularly in low- and middle-income countries.Objectives. To describe the clinical spectrum and outcomes associated with S. aureus infection in children &lt;14 years of age hospitalised atChris Hani Baragwanath Academic Hospital (CHBAH), South Africa, and to identify risk factors of invasive disease.Methods. A retrospective, descriptive study was conducted at CHBAH to define the spectrum of clinical presentation, risk factors,duration of treatment and outcomes of paediatric S. aureus infections for the period January through December 2013. Data were soughtfor all children &lt;14 years of age.Results. Four hundred and twenty-two episodes of S. aureus infection were identified. Three hundred and forty-two (81%) infectionswere caused by methicillin-susceptible S. aureus (MSSA) and 80 (19%) by methicillin-resistant S. aureus (MRSA). Clinical data wereobtained for 286 (67.8%) cases, on which all further analyses were based. Clinical presentations for MSSA bacteraemia included skin andsoft tissue infection (45%), pneumonia (10%), meningitis (6%), bone/joint infections (5%) and urinary tract infections (3%). Five (8.3%)cases of MRSA were community-acquired. Admission to intensive care unit (ICU) was the only risk factor associated with MRSA infection(adjusted odds ratio (aOR) 125.55; 95% confidence interval (CI) 11.67 - 1 350.68). Hospital-acquired S. aureus infection was the only factorassociated with mortality on multivariate analysis (aOR 8.70; 95% CI 1.55 - 48.77).Conclusion. S. aureus is frequently isolated in paediatric bacterial infections. Closer attention to infection control would impact on MRSAand S. aureus mortality rates in our setting.

Predicting Which Species of Bacteria Will Cause an Infection After Fracture Surgery.

The aim of this study was to develop and validate risk prediction models for deep surgical site infection (SSI) caused by specific bacterial pathogens after fracture fixation. A retrospective case-control study was conducted at a level I trauma center. Fifteen candidate predictors of the bacterial pathogens in deep SSI were evaluated to develop models of bacterial risk. The study included 441 patients with orthopedic trauma with deep SSI after fracture fixation and 576 control patients. The main outcome measurement was deep SSI cultures positive for methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant S aureus (MRSA), gram-negative rods (GNRs), anaerobes, or polymicrobial infection within 1 year of injury. Prognostic models were developed for five bacterial pathogen outcomes. Mean area under the curve ranged from 0.70 (GNRs) to 0.74 (polymicrobial). Strong predictors of MRSA were American Society of Anesthesiologists (ASA) classification of III or greater (odds ratio [OR], 3.4; 95% CI, 1.6-8.0) and time to fixation greater than 7 days (OR, 3.4; 95% CI, 1.9-5.9). Gustilo type III fracture was the strongest predictor of MSSA (OR, 2.5; 95% CI, 1.6-3.9) and GNRs (OR, 3.4; 95% CI, 2.3-5.0). ASA classification of III or greater was the strongest predictor of polymicrobial infection (OR, 5.9; 95% CI, 2.7-15.5) and was associated with increased odds of GNRs (OR, 2.7; 95% CI, 1.5-5.5). Our models predict the risk of MRSA, MSSA, GNR, anaerobe, and polymicrobial infections in patients with fractures. The models might allow for modification of preoperative antibiotic selection based on the particular pathogen posing greatest risk for this patient population. [Orthopedics. 2024;47(1):e19-e25.].

Integrated transcriptomic analysis reveals T and B cell immune signatures influencing resolving outcome in MRSA bacteremia

Abstract Staphylococcus aureus bacteremia is a potentially lethal infection particularly when involving methicillin-resistant S. aureus (MRSA). Contrary to MRSA bacteremia that resolves on its own (RB), persistent MRSA bacteremia (PB) happens when the infection persists after receiving antibiotic therapy. Host immune responses in PB vs. RB are poorly understood. Our goal was to identify proteomic, epigenetic, and transcriptomic features associated with PB and RB clinical outcomes. We examined the whole-blood transcriptomes of propensity score matched PB (n = 28) and RB (n = 30) patients. Gene expression (GE) modules were analysed by weighted gene co-expression network analysis (WGCNA) and quantified by module eigengene analysis. Enriched pathways and protein-protein interaction networks were identified. The correlation between host DNA methlytransferase-3A (DNMT3A) genotype and serum IL-10 level was examined in the identified GE modules. Upregulation of T and B cell immune response genes early during MRSA infection discriminate RB from PB patients. T and B cell signalling pathways were significantly increased in RB patients and correlated with lower serum IL-10 levels and the DNMT3A heterozygous A/C genotype. These findings indicate that patients with DNMT3A A/A genotypes have dysregulated adaptive immune responses characterized by impaired T and B cell function that increase the risk of PB outcome. These findings suggest that combining the genotype of the host with transcriptome and proteomic profiles might help determine the outcome of MRSA bacteremia. This knowledge may result in a better understanding of S. aureus pathogenesis and development of new anti-infective therapies and vaccines to improve patient outcomes. These studies were supported by in-part by the following NIH grants U01-AI124319, R01-AI068804 and R33-AI111661(to MRY) and U01-AI124319 and U19AL128913 (to EFR).