Neonatal methicillin-resistant Staphylococcus aureus pneumonia-related recurrent fatal pyopneumothorax: A case report and review of literature

BACKGROUND Although neonatal Staphylococcus aureus pneumonia is common and usually curable,it can also be refractory and life-threatening.Herein,we report a case of severe neonatal community-acquired methicillin-resistant Staphylococcus aureus(CA-MRSA)necrotizing pneumonia with bilateral recurrent pyopneumothorax,respiratory failure,heart failure,and cardiac arrest.We hope our report will add to the understanding of this disease.CASE SUMMARY An 18-d-old boy presented with cough for five days,fever for three days,and dyspnea for two days.Preadmission chest radiograph revealed high-density shadows in both lungs.On admission,his oxygen saturation fluctuated around 90%under synchronized intermittent mandatory ventilation.He was unconscious,with dyspnea,weak heart sounds and hepatomegaly.Moist crackles were present throughout his left lung,while the breath sounds in the right lung were decreased.After high-frequency oscillatory ventilation,empiric antimicrobials(meropenem and vancomycin),improved circulation,and right pleural cavity drainage for right pneumothorax(approximately 90%compression),his oxygen saturation level stayed above 95%,and recruitment of the right lung was observed.His condition did not deteriorate until the 5th day of hospitalization(DOH 5).On the morning of DOH 5,his oxygen saturation decreased.Subsequent chest radiograph showed bilateral pneumothorax with nearly 100%compression of the left lung.Desaturation was not relieved after urgent left pleural cavity drainage,and cardiac arrest occurred soon thereafter.Although his spontaneous heartbeat returned through emergency resuscitation and salvage antibacterial therapy(linezolid and levofloxacin)was administered given the detection and antimicrobial susceptibility of MRSA,he showed no improvement,with recurrent pyopneumothorax and continued drainage of purulent fluid and necrotic lung tissue fragments from the pleural cavity.Eventually,his parents refused extracorporeal membrane oxygenation(ECMO)and gave up all the treatments,and the newborn passed away soon after withdrawal on DOH 13.CONCLUSION Neonatal MRSA pneumonia can be refractory and lethal,especially in cases where necrotizing pneumonia leads to extensive lung necrosis and recurrent pneumothorax.Despite treatment with linezolid and other medical measures,it may still be ineffective.Currently,ECMO has been a remedial therapy,but if the lung tissue is too severely eroded to be repaired,it may be useless unless the infection can be controlled and lung transplantation can be performed.Regardless of whether ECMO is initiated,the key to successful treatment is to achieve control over the pneumonia caused by MRSA as soon as possible and to reverse lung injury as much as possible.

Elimination of methicillin‑resistant Staphylococcus aureus biofilms on titanium implants via photothermally‑triggered nitric oxide and immunotherapy for enhanced osseointegration

Background:Treatment of methicillin-resistant Staphylococcus aureus(MRSA)biofilm infections in implant placement surgery is limited by the lack of antimicrobial activity of titanium(Ti)implants.There is a need to explore more effective approaches for the treatment of MRSA biofilm infections.Methods:Herein,an interfacial functionalization strategy is proposed by the integration of mesoporous polydopamine nanoparticles(PDA),nitric oxide(NO)release donor sodium nitroprusside(SNP)and osteogenic growth peptide(OGP)onto Ti implants,denoted as Ti-PDA@SNP-OGP.The physical and chemical properties of Ti-PDA@SNP-OGP were assessed by scanning electron microscopy,X-ray photoelectron spectroscope,water contact angle,photothermal property and NO release behavior.The synergistic antibacterial effect and elimination of the MRSA biofilms were evaluated by 2′,7′-dichlorofluorescein diacetate probe,1-N-phenylnaphthylamine assay,adenosine triphosphate intensity,O-nitrophenyl-β-D-galactopyranoside hydrolysis activity,bicinchoninic acid leakage.Fluorescence staining,assays for alkaline phosphatase activity,collagen secretion and extracellular matrix mineralization,quantitative real‑time reverse transcription‑polymerase chain reaction,and enzyme-linked immunosorbent assay(ELISA)were used to evaluate the inflammatory response and osteogenic ability in bone marrow stromal cells(MSCs),RAW264.7 cells and their co-culture system.Giemsa staining,ELISA,micro-CT,hematoxylin and eosin,Masson's trichrome and immunohistochemistry staining were used to evaluate the eradication of MRSA biofilms,inhibition of inflammatory response,and promotion of osseointegration of Ti-PDA@SNP-OGP in vivo.Results:Ti-PDA@SNP-OGP displayed a synergistic photothermal and NO-dependent antibacterial effect against MRSA following near-infrared light(NIR)irradiation,and effectively eliminated the formed MRSA biofilms by inducing reactive oxygen species(ROS)-mediated oxidative stress,destroying bacterial membrane integrity and causing leakage of intracellular components(P<0.01).In vitro experiments revealed that Ti-PDA@SNP-OGP not only facilitated osteogenic differentiation of MSCs,but also promoted the polarization of pro-inflammatory M1 macrophages to the anti-inflammatory M2-phenotype(P<0.05 or P<0.01).The favorable osteo-immune microenvironment further facilitated osteogenesis of MSCs and the anti-inflammation of RAW264.7 cells via multiple paracrine signaling pathways(P<0.01).In vivo evaluation confirmed the aforementioned results and revealed that Ti-PDA@SNP-OGP induced ameliorative osseointegration in an MRSA-infected femoral defect implantation model(P<0.01).Conclusions:Ti-PDA@SNP-OGP is a promising multi-functional material for the high-efficient treatment of MRSA infections in implant replacement surgeries.

Genotypic Characterization of Methicillin-resistant Staphylococcus aureus Isolated from Pigs and Retail Foods in China

Objective To investigate the genotypic diversity of Methicillin-resistant Staphylococcus aureus （MRSA） isolated from pigs and retail foods from different geographical areas in China and further to study the routes and rates of transmission of this pathogen from animals to food. Methods Seventy-one MRSA isolates were obtained from pigs and retail foods and then characterized by multi-locus sequencing typing （MLST）, spa typing, multiple-locus variable number of tandem repeat analysis （MLVA）, pulsed-field gel electrophoresis （PFGE）, and antimicrobial susceptibility testing. Results All isolated MRSA exhibited multi-drug resistance （MDR）. Greater diversity was found in food-associated MRSA （7 STs, 8 spa types, and 10 MLVA patterns） compared to pig-associated MRSA （3 STs, 1 spa type, and 6 MLVA patterns）. PFGE patterns were more diverse for pig-associated MRSA than those of food-associated isolates （40 vs. 11 pulse types）. Among the pig-associated isolates, CC9-ST9-t899-MC2236 was the most prevalent clone （96.4%）, and CC9-ST9-t437-MC621 （20.0%） was the predominant clone among the food-associated isolates. The CC9-ST9 isolates showed significantly higher antimicrobial resistance than other clones. Interestingly, CC398-ST398-t034 clone was identified from both pig- and food-associated isolates. Of note, some community- and hospital-associated MRSA strains （t030, t172, t1244, and t4549） were also identified as food-associated isolates. Conclusion CC9-ST9-t899-MC2236-MDR was the most predominant clone in pigs, but significant genetic diversity was observed in food-associated MRSA. Our results demonstrate the great need for improved surveillance of MRSA in livestock and food and effective prevention strategies to limit MDR-MRSA infections in China.

Potentiating activity of luteolin on membrane permeabilizing agent and ATPase inhibitor against methicillin-resistant Staphylococcus aureus

Objective:To investigate the mechanism of antibacterial activity of luteoiin(LUT) against methicillin-resistant Staphylococcus aureus(MRSA).Methods:The mechanism of anti-MRSA activity of LUT was analyzed by the viability assay in membrane permeabilizing agent ATPase inhibitors,and peptidoglycan(PGN) derived from Staphylococcus aureus(S.aureus).Also,transmission electron microscopy was used to monitor survival characteristics and changes in S.aureus morphology.Results:Compared to the LUT alone,the optical density of suspensions treated with the combination of 125 μg/mL Tris and 230 μg/mL DCCD were reduced to 60%and 46%,respectively.PGN(15.6 μg/mL) gradually impeded the activity of LUT,and PGN(62.5 μg/mL) completely blocked the activity of LUT on S.aureus.Conclusions:Increased susceptibility to LUT with me Tris and DCCD combinations is evident in all tested MRSA isolates.The results indicate LUT synergy in increasing cytoplasmic membrane permeability and inhibiting ATPase.S.aureus PGN directly blocks the antibacterial activity of LUT,suggesting the direct binding of LUT with PGN.These findings may be validated for the development of antibacterial agent for low MRSA resistance.

Postoperation of preauricular fistula cellulitis caused by methicillin-resistant staphylococcus aureus infection

A one-year-old baby girl with one-month history of recurrent pus fluid exuding from her left preauricular sinus orifice, who failed multiple courses of surgical drainage of the abscess and persistent debridement for the wound, presented with MRSA infection. The patient was treated with linezolid for three days. Her pain and paresthesia resolved, and C-reactive protein decreased to normal.

Teicoplanin combined with conventional vancomycin therapy for the treatment of pulmonary methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis infections

BACKGROUND Vancomycin and teicoplanin are both antibiotics that have significant antimicrobial effects on Gram-positive cocci.AIM To explore the value of teicoplanin combined with conventional(vancomycin only)anti-infective therapy for the treatment of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis pulmonary infections.METHODS A total of 86 patients with methicillin-resistant Staphylococcus aureus or methicillin-resistant Staphylococcus epidermidis pulmonary infections,treated in our hospital between January 2018 and February 2020,were assigned to the study and control groups using a random number table method,with 43 patients in each group.The control group received conventional treatment(vancomycin),and the study group received both teicoplanin and conventional treatment.The following indicators were assessed in both groups:the time required for symptom relief,treatment effectiveness,serum levels of inflammatory factors(procalcitonin,interleukin-1β,tumor necrosis factor-α,C-reactive protein),clinical pulmonary infection scores before and after treatment,and the incidence of adverse reactions.RESULTS Patients in the study group were observed to have faster cough and expectoration resolution,white blood cell count normalization,body temperature normalization,and rales disappearance than patients in the control group(all P<0.05);the total rate of effectiveness was 93.02%in the study group,higher than the 76.74%in the control group(P<0.05).The pre-treatment serum levels of procalcitonin,interleukin-1β,tumor necrosis factor-α,and C-reactive protein as well as the clinical pulmonary infection scores were similar among the patients in both groups.However,the post-treatment serum levels of procalcitonin,interleukin-1β,tumor necrosis factor-α,and C-reactive protein as well as the clinical pulmonary infection scores were significantly lower in the study group than in the control group(P<0.05).There was no significant difference in the incidence of adverse reactions between the groups.CONCLUSION Compared with conventional(vancomycin only)therapy,teicoplanin and vancomycin combination therapy for patients with pulmonary methicillinresistant Staphylococcus aureus and methicillin-resistant Staphylococcus epidermidis infections can improve patient clinical symptoms,modulate serum inflammatory factor levels,and improve treatment efficacy,without increasing the risk of adverse reactions.

Antimicrobial activity and synergism of Sami-Hyanglyun-Hwan with ciprofloxacin against methicillin-resistant Staphylococcus aureus

Objective: To investigate the antibacterial activity of SHHextracted with either water or ethanol against methicillin-resistant Staphylococcus aureus(MRSA) and combinatory antimicrobial effect with ciprofloxacin(CIP) by time kill assay and checkerboard dilution test. Methods: The antibacterial activity determined by broth dilution method indicated that the antibacterial activity of Sami-Hyanglyun-Hwan(SHH) water extract(SHHW) and SHH ethanol extract(SHHE) ranged from 250 to 2000 μg/m L and 125 to 1000 μg/m L against MRSA, respectively. Results: In the checkerboard method, the combinations of SHHE with CIP had a partial synergistic or synergistic effect against MRSA. The time-kill curves showed that a combined SHHE and CIP treatment reduced the bacterial counts dramatically after 24 h. Conclusions: The present study demonstrates the therapeutic ability of SHHE against MRSA infections.

Surface-engineered liposomes for dual-drug delivery targeting strategy against methicillin-resistant Staphylococcus aureus(MRSA)

This study focused on the encapsulation of vancomycin(VAN) into liposomes coated with a red blood cell membrane with a targeting ligand, daptomycin–polyethylene glycol–1,2-distearoyl-sn-glycero-3-phosphoethanolamine, formed by conjugation of DAPT and Nhydroxysuccinimidyl-polyethylene glycol-1,2-distearoyl-sn-glycero-3-phosphoethanolamine.This formulation is capable of providing controlled and targeted drug delivery to the bacterial cytoplasm. We performed MALDI-TOF, NMR and FTIR analyses to confirm the conjugation of the targeting ligand via the formation of amide bonds. Approximately 45% of VAN could be loaded into the aqueous cores, whereas 90% DAPT was detected using UV–vis spectrophotometry. In comparison to free drugs, the formulations controlled the release of drugs for > 72 h. Additionally, as demonstrated using CLSM and flow cytometry, the resulting formulation was capable of evading detection by macrophage cells. In comparison to free drugs, red blood cell membrane–DAPT–VAN liposomes, DAPT liposomes, and VAN liposomes reduced the MIC and significantly increased bacterial permeability, resulting in > 80% bacterial death within 4 h. Cytotoxicity tests were performed in vitro and in vivo on mammalian cells,in addition to hemolytic activity tests in human erythrocytes, wherein drugs loaded into the liposomes and RBCDVL exhibited low toxicity. Thus, the findings of this study provide insight about a dual antibiotic targeting strategy that utilizes liposomes and red blood cell membranes to deliver targeted drugs against MRSA.

Daptomycin and linezolid for severe methicillin-resistant Staphylococcus aureus psoas abscess and bacteremia:A case report and review of the literature

BACKGROUND Vancomycin remains a first-line treatment drug as per the treatment guidelines for methicillin-resistant Staphylococcus aureus(MRSA)bacteremia.However,a number of gram-positive cocci have developed resistance to several drugs,including glycopeptides.Therefore,there is an urgent need for effective and innovative antibacterial drugs to treat patients with infections caused by drugresistant bacteria.CASE SUMMARY A 24-year-old male was admitted to hospital owing to lumbago,fever,and hematuria.Computed tomography(CT)results showed an abscess in the psoas major muscle of the patient.Repeated abscess drainage and blood culture suggested MRSA,and vancomycin was initiated.However,after day 10,CT scans showed abscesses in the lungs and legs of the patient.Therefore,treatment was switched to daptomycin.Linezolid was also added considering inflammation in the lungs.After 10 d of the dual-drug anti-MRSA treatment,culture of the abscess drainage turned negative for MRSA.On day 28,the patient was discharged without any complications.CONCLUSION This case indicates that daptomycin combined with linezolid is an effective remedy for bacteremia caused by MRSA with pulmonary complications.

Synergistic Effect between Fucoidan and Antibiotics against Clinic Methicillin-Resistant <i>Staphylococcus aureus</i>

Fucoidan is a sulfated polysaccharide that is primarily extracted from brown seaweeds which has been broadly studied in recent years due to its numerous biological properties, including anticoagulant, antithrombotic, antitumor, and antiviral activities. In this study, fucoidan was evaluated against clinic isolated methicillin-resistant Staphylococcus aureus (MRSA) 1 - 20, either alone or with antibiotics, via broth dilution method and checkerboard and time kill assay. Minimum inhibitory concentrations (MICs)/Minimum bactericidal concentrations (MBCs) values for the fucoidan against all the tested bacteria ranged between 64 - 512/256 - 2048 microg/mL, for ampicillin 32 - 1024/64 - 1024 microg/mL and for oxacillin 8 - 64/16 - 256 microg/mL respectively. Furthermore, the MIC and MBC were reduced to one half-eighth as a result of the combination of the fucoidan with antibiotics. 2 - 6 hours of treatment with 1/2 MIC of fucoidan with 1/2 MIC of antibiotics resulted from an increase of the rate of killing in units of CFU/mL to a greater degree than was observed with alone. These results suggest that fucoidan could be employed as a natural antibacterial agent against multi-drug bacteria.

Treatment of methicillin-resistant Staphylococcus aureus infections:Importance of high vancomycin minumum inhibitory concentrations

Staphylococcus aureus(SA) infections remain a major cause of morbidity and mortality despite the availability of numerous effective anti-staphylococcal antibiotics.This organism is responsible for both nosocomial and community-acquired infections ranging from relatively minor skin and soft tissue infections to life-threateningsystemic infections.The increasing incidence of methicillin-resistant strains has granted an increasing use of vancomycin causing a covert progressive increase of its minimum inhibitory concentration(MIC)(dubbed the MIC "creep").In this way,the emergence of vancomycinintermediate SA(VISA) strains and heteroresistantVISA has raised concern for the scarcity of alternative treatment options.Equally alarming,though fortunately less frequent,is the emergence of vancomycin-resistant SA.These strains show different mechanisms of resistance but have similar problems in terms of therapeutic approach.Ultimately,various debate issues have arisen regarding the emergence of SA strains with a minimum inhibitory concentration sitting on the superior limit of the sensitivity range(i.e.,MIC = 2 μg/mL).These strains have shown certain resilience to vancomycin and a different clinical behaviour regardless of vancomycin use,both in methicillin-resistant SA and in methicillin-sensitive SA.The aim of this text is to revise the clinical impact and consequences of the emergence of reduced vancomycin susceptibility SA strains,and the different optimal treatment options known.

The prevalence of methicillin-resistant Staphylococcus aureus colonization in emergency department fast track patients

BACKGROUND:Over the past two decades,methicillin-resistant Staphylococcus aureus(MRSA) has evolved from a hospital-associated infection to a significant public health threat in the community,causing outbreaks of soft tissue infections in otherwise healthy individuals.The goal of this study was to determine the prevalence of nasal MRSA colonization in low acuity Emergency Department(ED) Fast Track patients in order to better characterize the epidemiology of this pathogen.METHODS:We conducted a cross-sectional study of a convenience sample of adult patients from our ED Fast Track.Nasal swabs were analyzed for MRSA using a polymerase chain reaction assay.Study participants completed a survey assessing traditional risk factors for CA-MRSA colonization.RESULTS:A total of 106 ED Fast Track patients were tested.Four(3.8%,95%CI 1.5%-9.3%)were MRSA positive.Three traditional CA-MRSA risk factors(personal history of abscess,family history of abscess,and participation in contact sports) were examined.In patients with a positive MRSA nasal swab,only a personal prior history of abscess retained significance(OR 33,95%C11.7-676,P=0.02).CONCLUSION:This study found a higher prevalence of nasal MRSA colonization in low acuity ED Fast Track patients compared with historical community surveillance studies.A personal history of prior abscess was a significant risk for CA-MRSA carriage.

Crohn's disease complicated by intestinal infection with methicillin-resistant Staphylococcus aureus

We report on a 24-year-old male patient with history of bloody diarrhea, abdominal pain and vomiting. Endoscopy revealed massive ulcerative discontinuous proctosigmoiditis with deep, sharply demarcated epithelial denudations and enterotoxigenic methicillin-resistant Staphylococcus aureus (MRSA) was detected in mucosal biopsies. After treatment with linezolide and steroids, a significant amelioration of colitis was detected and testing for MRSA became negative. In face of the case presented here, we suggest that in patients with refractory inflammatory bowel disease (IBD), microbiological assessment should be performed to detect a possible Staphylococcus aureus infection in order to initiate an antimicrobial treatment in addition to IBDspecific treatment.

Molecular study on methicillin-resistant Staphylococcus aureus strains isolated from dogs and associated personnel in Jordan

Objective: To determine the prevalence, genetic relatedness, and pattern of antimicrobial susceptibility in methicillin-resistant Staphylococcus aureus(S. aureus)(MRSA) isolated from household dogs, farm dogs, and stray dogs, compared to isolates from their associated personnel.Methods: MRSA was isolated from 250 nasal swabs(150 swabs from dogs and 100 swabs from humans). PCR assays were used to detect the presence of both the nuc and mec A genes,which con firmed the identity of S. aureus isolates and the presence of methicillin resistance,respectively. Disk diffusion was used to determine the antibiotic susceptibility against 15 antimicrobial agents along with an E-test that determined the minimum inhibitory concentration for oxacillin. Pulsed field gel electrophoresis was conducted to determine the genetic relatedness of MRSA isolates from dogs to those from associated and unassociated personnel.Results: The prevalence of S. aureus in dogs and humans was 12.7% and 10.0%respectively, while the prevalence of MRSA isolates in dogs and humans was 5.3% and5.0%, respectively. The prevalence of MRSA isolates in household dogs, farm dogs, and stray dogs was 7.8%, 4.7%, and 0.0%, respectively. MRSA isolates demonstrated a significantly higher rate of multi-resistance against three or more antimicrobial agents than methicillin-susceptible S. aureus(MSSA). Trimethoprim-sulphamethoxazole and chloramphenicol were the most effective antibiotics against all MRSA isolates. Pulsed field gel electrophoresis revealed a strong association between dog MRSA isolates and MRSA isolates from strongly associated personnel.Conclusions: MRSA is prevalent in house dogs, as well as in dog rearing centers and among their strongly associated personnel. A strong association was found between the MRSA isolates from dogs and those from humans who are in close contact. In addition,MRSA isolates showed a high rate of multi-resistance compared to MSSA isolates.

A case of methicillin-resistant Staphylococcus aureus infection following bile duct stenting

AIM: To present a case of methicillin-resistant Staphylococcus aureus (MRSA) infection following bile duct stenting in a patient with malignant biliary obstruction.METHODS: A 78-year-old male patient was admitted to a community hospital with progredient painless jaundice lasting over two weeks, weight loss and sweating at night.Whether a stent should be implanted pre-operatively in jaundiced patients or whether these patients should directly undergo surgical resection, was discussed.RESULTS: ERC and a biopsy from the papilla of Vater revealed an adenocarcinoma. In addition, a 7-Ch plastic stent was placed into the common bile duct. Persistent abdominal pain, increasing jaundice, weakness and indigestion led to the transfer of the patient to our hospital.A pylorus-preserving pancreatoduodenectomy wasperformed. Intraoperatively, bile leaked out of the transected choledochus andthe stent was found to be dislocated in the duodenum. A smear of the bile revealed an infection with MRSA, leading to post-operative isolation of the patient.CONCLUSION:As biliary stents can cause severe infection of the bile, the need for pre-operative placement of biliary stents should be carefully evaluated in each individual case.

Emergence of staphylococcal cassette chromosome mec type Ⅰ with high-level mupirocin resistance among methicillin-resistant Staphylococcus aureus

Objective: To investigate the molecular epidemiology and antimicrobial resistance patterns of methicillin-resistant Staphylococcus aureus(MRSA) among healthcare workers and patients.Methods: MRSA isolates were recovered from nasal swabs collected at a tertiary care hospital of Nepal and confirmed on the basis of Gram staining, conventional biochemical tests, and PCR amplification of mec A gene. PCRs were also used for detection of the different resistance genes and staphylococcal cassette chromosome(SCC) mec types.Antibiotic susceptibility patterns of isolates were assessed by disc diffusion method and minimum inhibitory concentrations were determined by E-test.Results: A total of 29 MRSA were isolated from 536 nasal swabs(5.4%) of health care workers and patients at a tertiary care hospital in Nepal. All isolates were susceptible to amikacin, gentamicin, vancomycin(minimal inhibitory concentrations < 2 mg/m L), tigecycline, tetracycline, nitrofurantoin, rifampicin, quinupristin-dalfopristin, and linezolid. Among the 29 MRSA isolates, resistance to erythromycin(72%), ciprofloxacin(75%), co-trimoxazole(62%), clindamycin(10%), and chloramphenicol(10%) was found, and fifteen isolates(51%)exhibited high-level mupirocin resistance(minimal inhibitory concentrations > 1 024 mg/m L).Fourteen isolates were found harboring the mup A gene and one isolate was found carrying the novel mup B gene. High prevalence(68%) of SCCmec I type was found, followed by SCCmec V(13%) and SCCmec III(3%) among all the MRSA isolates.Conclusions: We found the emergence of SCCmec type I with high-level mupirocin resistance among MRSA in Nepal. Data also suggest that MRSA SCCmec type V strain has spread from the community to the hospital.

Screening agars for MRSA:evaluation of a stepwise diagnostic approach with two different selective agars for the screening for methicillin-resistant Staphylococcus aureus(MRSA)

Background: Colonization with methicillin-resistant Staphylococcus aureus(MRSA) poses a hygiene risk that does not spare field hospitals or military medical field camps during military deployments. Diagnostic options for unambiguously identifying MRSA isolates are usually scarce in military environments. In this study, we assessed the stepwise application of two different selective agars for the specific identification of MRSA in screening analyses.Methods: Nasal swabs from 1,541 volunteers were subjected to thioglycollate broth enrichment and subsequently screened on CHROMagar MRSA selective agar for the identification of MRSA. The MRSA identity of suspiciouslooking colonies was confirmed afterwards or excluded by another selective agar, chrom ID MRSA. All isolates from the selective agars with MRSA-specific colony morphology were identified by biochemical methods and mass spectrometry.Results: The initial CHROMagar MRSA screening identified suspicious colonies in 36 out of 1541 samples. A total of 25 of these 36 isolates showed MRSA-like growth on chrom ID agar. Out of these 25 isolates, 24 were confirmed as MRSA, while one isolate was identified as Staphylococcus kloosii. From the 11 strains that did not show suspicious growth on chrom ID agar, 3 were methicillin-sensitive Staphylococcus aureus(MSSA, with one instance of cocolonization with Corynebacterium spp.), 2 were confirmed as MRSA(with 1 instance of co-colonization with MSSA), 2 were lost during passaging and could not be re-cultured, one could not be identified by the applied approaches, and the remaining 3 strains were identified as Staphylococcus saprophyticus, Staphylococcus hominis(co-colonized with Macrococcus caseolyticus) and Staphylococcus cohnii, respectively.Conclusion: The application of the selective agar CHROMagar MRSA alone proved to be too non-specific to allow for a reliable diagnosis of the presence of MRSA. The combined use of two selective agars in a stepwise approach reduced this non-specificity with an acceptably low loss of sensitivity. Accordingly, such a stepwise screening approach might be an option for resource-restricted military medical field camps.

Inhibition mechanisms of secretome proteins from Paenibacillus polymyxa Kp10 and Lactococcus lactis Gh1 against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus

Objective:To determine the inhibition mechanisms of secretome protein extracted from Paenibacillus polymyxa Kp10(Kp10)and Lactococcus lactis Gh1(Gh1)against methicillin-resistant Staphylococcus aureus(MRSA)and vancomycin-resistant Enterococcus(VRE).Methods:The sensitivity and viability of MRSA and VRE treated with secretome proteins of Kp10 and Gh1 were determined using minimal inhibitory concentration,minimum bactericidal concentration,and time-to-kill assays.The morphological changes were observed using scanning electron microscopy and transmission electron microscopy.To elucidate the antimicrobial mechanism of secretome protein of Kp10 and Gh1 against MRSA and VRE,2D gel proteomic analysis using liquid chromatography-mass spectrometry was run by comparing upregulated and downregulated proteins,and the proton motive force study including the efflux of ATP,pH gradient,and the membrane potential study were conducted.Results:MRSA and VRE were sensitive to Kp10 and Gh1 secretome protein extracts and displayed apparent morphological and internal composition changes.Several proteins associated with cellular component functions were either downregulated or upregulated in treated MRSA and VRE by changing the membrane potential gradient.Conclusions:Kp10 and Gh1 secretome proteins reduce the growth of VRE and MRSA by damaging the cell membrane.Cell division,cell wall biosynthesis,and protein synthesis are involved in the inhibition mechanism.

Methicillin-resistant Staphylococcus aureus infected gluteal compartment syndrome with rhabdomyolysis in a bodybuilder

Gluteal compartment syndrome(GCS) is a rare condition. We present a case of gluteal muscle strain with hematoma formation, methicillin-resistant Staphylococcus aureus(MRSA) superinfection, leading to acute GCS, rhabdomyolysis and acute kidney injury. This combination of diagnoses has not been reported in the literature. A 36-year-old Caucasian male presented with buttock pain, swelling and fever after lifting weights. Gluteal compartment pressure was markedly elevated compared with the contralateral side. Investigations revealed elevated white blood cell, erythrocyte sedimentation rate, C-reactive protein, creatine kinase, creatinine and lactic acid. Urinalysis was consistent with myoglobinuria. Magnetic resonance imaging showed increased T2 signal in the gluteus maximus and a central hematoma. Cultures taken from the emergency debridement and fasciotomy revealed MRSA. He had repeat, debridement 2 d later, and delayed primary closure 3 d after. GCS is rare and must be suspected when patients present with pain and swelling after an inciting event. They are easily diagnosed with compartment pressure monitoring. The treatment of gluteal abscess and compartment syndrome is the same and involves rapid surgical debridement.

Sanchen powder extract combined with vancomycin against methicillin-resistant Staphylococcus aureus

Objective: To explore the effects of the extract of Sanchen powder(ESCP) combined with vancomycin on methicillin-resistant Staphylococcus aureus(MRSA) planktonic cells, biofilms, and virulence factors.Methods: The herbs in Sanchen powder(SCP) were extracted separately with 50% ethanol. Then, the content of hydroxysafflower yellow A(HSYA) and cholic acid in the extract of artificial cattle bezoar(artificial Calculus bovis) and safflower(Carthamus tinctorius L.) was measured by ultraviolet visible-vis spectrophotometry and high-performance liquid chromatography. The effects of ESCP combined with vancomycin on MRSA by observing its biofilm viability were assessed using a 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium5-carboxanilide reduction assay and scanning electron microscopy.Additionally, enzyme activity was measured by plasma coagulase test and DNase test.Results: The cholic acid content of the artificial C. bovis extract was 7.34(0.81) mg/g, and the HSYA content of the C. tinctorius extract was 9.18(0.09) mg/g. The minimal inhibitory concentrations of ESCP and vancomycin were 25.6 mg/m L and 2 μg/m L. The minimum bactericidal concentration of ESCP was51.2 mg/m L. ESCP combined with vancomycin could inhibit the expression of coagulase and bacteria in mature biofilms. Neither ESCP nor vancomycin had a significant effect on DNase.Conclusion: This study is the first to show that ESCP combined with vancomycin inhibits coagulase and MRSA embedded in mature biofilms and that it represents a promising treatment for MRSA infection.