Objective: To analyze the Staphylococcal enterotoxins, Staphylococcal enterotoxin genes, drug resistance and molecular typing of 41 Staphylococcus aureus isolates from 2 food-borne illness outbreaks on 21 August and 27 September 2020 in Guangzhou. Methods: A total of 41 Staphylococcus aureus isolates from 2 outbreaks were analyzed by multilocus sequence typing (MLST) and spa typing. The Staphylococcal enterotoxins typing and the Staphylococcal enterotoxin genes of the isolates were analyzed by ELISA and PCR, respectively. The antimicrobial susceptibility of the isolates was performed by disc diffusion. 21 Staphylococcus aureus isolates were characterized using whole genome sequencing (WGS). Based on the whole genome single nucleotide polymorphism (SNP), the phylogenetic tree was constructed by Snippy. Results: 41 Staphylococcus aureus isolates were divided into 2 types by MLST and spa typing: ST6-t701 and ST7-t091. 2 ST7-t091 isolates were identified as methicillin-resistant Staphylococcus aureus (MRSA). 25 ST7-t091 isolates and 14 ST6-t701 isolates were methicillin-sensitive Staphylococcus aureus (MSSA), and were resistant to 7 and 6 antibiotics, respectively. All isolates were positive for sea by PCR. WGS revealed all 21 isolates carried scn, sak, sea, hla, hld, hlgA, hlgB, hlgC, lukD virulence genes. The results showed the isolates contained an immune evasion cluster type D which located in bacteriophage ϕSa3. The SNP phylogenetic tree showed 2 MRSA ST7-t091 were constituted a separate clade from the 12 MSSA ST7-t091 isolates and 7 ST6-t701 isolates showed high similarity to each other. Conclusion: Base on the results of phylogenetic analysis, the 2 food-borne illness outbreaks occurred on 21 August and 27 September 2020 are caused by the combination of the MRSA ST7-t091 strain and the MSSA ST7-t091 strain, and the MSSA ST6-t701 strain, respectively. All isolates have high level of antibiotic resistance and carry high virulent genes.
Anti-Bacterial Agents/pharmacology* ; Disease Outbreaks ; Foodborne Diseases/epidemiology* ; Humans ; Methicillin-Resistant Staphylococcus aureus/genetics* ; Microbial Sensitivity Tests ; Multilocus Sequence Typing/methods* ; Phylogeny ; Staphylococcal Infections/epidemiology* ; Staphylococcus aureus/genetics*

Emerging antibiotic resistance is a major global health threat. The analysis of nucleic acid sequences linked to susceptibility phenotypes facilitates the study of genetic antibiotic resistance determinants to inform molecular diagnostics and drug development. We collected genetic data (11,087 newly-sequenced whole genomes) and culture-based resistance profiles (10,991 out of the 11,087 isolates comprehensively tested against 22 antibiotics in total) of clinical isolates including 18 main species spanning a time period of 30 years. Species and drug specific resistance patterns were observed including increased resistance rates for Acinetobacter baumannii to carbapenems and for Escherichia coli to fluoroquinolones. Species-level pan-genomes were constructed to reflect the genetic repertoire of the respective species, including conserved essential genes and known resistance factors. Integrating phenotypes and genotypes through species-level pan-genomes allowed to infer gene-drug resistance associations using statistical testing. The isolate collection and the analysis results have been integrated into GEAR-base, a resource available for academic research use free of charge at https://gear-base.com.
Acinetobacter baumannii ; genetics ; isolation & purification ; Bacteria ; genetics ; isolation & purification ; Cell Culture Techniques ; methods ; Drug Resistance, Microbial ; genetics ; Escherichia coli ; genetics ; isolation & purification ; Genome, Bacterial ; Genotype ; Humans ; Internet ; Microbial Sensitivity Tests ; Phenotype ; Whole Genome Sequencing

BACKGROUND: The emergence of carbapenem resistance among gram-negative bacilli (GNB), mediated by carbapenemase production, has necessitated the development of a simple and accurate device for detecting minimum inhibitory concentrations (MICs) and resistance mechanisms, especially carbapenemase production. We evaluated the performance of the BD Phoenix NMIC-500 panel (BD Diagnostic Systems, Sparks, MD, USA) for antimicrobial susceptibility testing (AST) and carbapenemase-producing organism (CPO) detection. METHODS: We used 450 non-duplicate clinical GNB isolates from six general hospitals in Korea (409 Enterobacteriaceae and 41 glucose non-fermenting bacilli [GNFB] isolates). AST for meropenem, imipenem, ertapenem, ceftazidime, and ceftazidime/avibactam, and CPO detection were performed using the Phoenix NMIC-500 panel. Broth microdilution was used as the reference method for AST. The rates of categorical agreement (CA), essential agreement (EA), minor error (mE), major error (ME), and very major error (VME) were calculated in each antimicrobial. In addition, PCR and sequencing were performed to evaluate the accuracy of CPO detection by the BD Phoenix NMIC-500 panel, and the rate of correct identification was calculated. RESULTS: The CA rates were >90% for all antimicrobials tested with the Enterobacteriaceae isolates, except for imipenem (87.2%). The GNFB CA rates ranged from 92.7% to 100% for all antimicrobials. The ME rates were 1.7% for Enterobacteriaceae and 0% for GNFB. The panel identified 97.2% (243/250) of the carbapenemase-producing isolates. CONCLUSIONS: The BD Phoenix NMIC-500 panel shows promise for AST and CPO detection.
Ceftazidime ; Drug Resistance, Bacterial ; Enterobacteriaceae ; Glucose ; Hospitals, General ; Imipenem ; Korea ; Methods ; Microbial Sensitivity Tests ; Polymerase Chain Reaction

OBJECTIVES: Irrigants are imperative in endodontic therapy for the elimination of pathogens from the infected root canal. The present study compared the antimicrobial efficacy of octenidine dihydrochloride (OCT) with chlorhexidine (CHX) and sodium hypochlorite (NaOCl) against Staphylococcus epidermidis (S. epidermidis) for root canal disinfection. MATERIALS AND METHODS: The minimum inhibitory concentration (MIC) was obtained using serial dilution method. The agar diffusion method was then used to determine the zones of inhibition for each irrigant. Lastly, forty 6-mm dentin blocks were prepared from human mandibular premolars and inoculated with S. epidermidis. Samples were randomly divided into 4 groups of 10 blocks and irrigated for 3 minutes with saline (control), 2% CHX, 3% NaOCl, or 0.1% OCT. Dentin samples were then collected immediately for microbial analysis, including an analysis of colony-forming units (CFUs). RESULTS: The MICs of each tested irrigant were 0.05% for CHX, 0.25% for NaOCl, and 0.0125% for OCT. All tested irrigants showed concentration-dependent increase in zones of inhibition, and 3% NaOCl showed the largest zone of inhibition amongst all tested irrigants (p < 0.05). There were no significant differences among the CFU measurements of 2% CHX, 3% NaOCl, and 0.1% OCT showing complete elimination of S. epidermidis in all samples. CONCLUSIONS: This study showed that OCT was comparable to or even more effective than CHX and NaOCl, demonstrating antimicrobial activity at low concentrations against S. epidermidis.
Agar ; Anti-Bacterial Agents ; Bicuspid ; Chlorhexidine ; Dental Pulp Cavity ; Dentin ; Diffusion ; Disinfection ; Humans ; In Vitro Techniques ; Methods ; Microbial Sensitivity Tests ; Sodium Hypochlorite ; Staphylococcus epidermidis ; Staphylococcus ; Stem Cells

OBJECTIVES: Staphylococcus aureus is a nosocomial pathogen that provides a major challenge in the healthcare environment, especially in burns units where patients are particularly susceptible to infections. In this study, we sought to determine molecular types of S. aureus isolates collected from burns patients, based on staphylococcal protein A and coagulase gene polymorphisms. METHODS: Antibiotic susceptibility testing of 89 S. aureus strains isolated from burn wounds of patients was assessed using the Kirby-Bauer disk diffusion method. Strains were characterized by spa typing, coa typing, and resistance and toxin gene profiling. RESULTS: A total of 12 different spa types were identified with the majority being t790 (18%). Panton-Valentine leucocidin encoding genes were identified in spa types t044 (5.6%), t852 (2.2%) and t008 (2.2%). The most commonly detected antibiotic resistance gene was ant (4′)-Ia (60.7%). Ten different coa types were detected and the majority of the tested isolates belonged to coa III (47.2%). All the high-level mupirocin-resistant and low-level mupirocin resistant strains belonged to coa type III. CONCLUSION: The present study illustrated that despite the high frequency of coa III and spa t790 types, the genetic background of S. aureus strains in Iranian burns patients was diverse. The findings obtained are valuable in creating awareness of S. aureus infections within burns units.
Ants ; Burns ; Coagulase ; Delivery of Health Care ; Diffusion ; Drug Resistance, Microbial ; Genetic Background ; Humans ; Leukocidins ; Methicillin Resistance ; Methicillin ; Methicillin-Resistant Staphylococcus aureus ; Methods ; Microbial Sensitivity Tests ; Mupirocin ; Staphylococcal Protein A ; Staphylococcus aureus ; Staphylococcus ; Wounds and Injuries

Anthrax, caused by Bacillus anthracis, is a non-contagious infectious disease that affects a wide range of animal species (primarily ruminants) including humans. Due to the often-fatal outcome in humans, quick administration of definitely effective antimicrobials is crucial either as prophylaxis or as a clinical case therapy. In this study, 110 B. anthracis strains, temporally, geographically, and genetically different, isolated during anthrax outbreaks in Italy from 1984 to 2017, were screened using a broth microdilution method to determine their susceptibility to 16 clinically relevant antimicrobial agents. The strains were isolated from various matrices (human, animal, and environmental samples) and were representative of thirty distinct genotypes previously identified by 15-loci multiple-locus variable-number of tandem repeats analysis. The antimicrobials tested were gentamicin, ceftriaxone, streptomycin, penicillin G, clindamycin, chloramphenicol, vancomycin, linezolid, cefotaxime, tetracycline, erythromycin, rifampin, amoxicillin, ciprofloxacin, doxycycline, and trimethoprim. All isolates were susceptible to most of the tested antimicrobials, with the exception of trimethoprim for which all of them showed high minimal inhibitory concentration values. An intermediate level of susceptibility was recorded for ceftriaxone and cefotaxime. Although the Centers for Disease Control and Prevention recommend the use of doxycycline, ciprofloxacin, penicillin G, and amoxicillin for treatment of human cases and for post-exposure prophylaxis to anthrax spores, this study shows a high degree of in vitro susceptibility of B. anthracis to many other antimicrobials, suggesting the possibility of an alternative choice for prophylaxis and therapy.
Amoxicillin ; Animals ; Anthrax ; Anti-Infective Agents ; Bacillus anthracis ; Bacillus ; Cefotaxime ; Ceftriaxone ; Centers for Disease Control and Prevention (U.S.) ; Chloramphenicol ; Ciprofloxacin ; Clindamycin ; Communicable Diseases ; Disease Outbreaks ; Doxycycline ; Erythromycin ; Genotype ; Gentamicins ; Humans ; In Vitro Techniques ; Italy ; Linezolid ; Methods ; Microbial Sensitivity Tests ; Penicillin G ; Post-Exposure Prophylaxis ; Rifampin ; Spores ; Streptomycin ; Tandem Repeat Sequences ; Tetracycline ; Trimethoprim ; Vancomycin

The aim of this study was to investigate the susceptibility of Mutans streptococci in both planktonic and biofilm states to erythrosine.S. mutans was cultured in brain-heart infusion (BHI) broth. Erythrosine was diluted in BHI broth and prepared at a concentration range of 0.02 – 10000 µg/L. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were measured using the microdilution method. After forming biofilms on 96-well plates, the minimum biofilm inhibitory concentration (MBIC) and minimum biofilm eradication concentration (MBEC) were measured.S. mutans was susceptible to erythrosine in both planktonic and biofilm states. MIC and MBC values were both 19.5 µg/L for the planktonic state, while MBIC and MBEC values were 313 µg/L and 2500 µg/L, respectively, for the biofilm state.Erythrosine (19.5 µg/L) exhibited a bactericidal effect on S. mutans (killing 99.9%) in the planktonic state. For biofilms, erythrosine inhibited biofilm growth and eradicated 99.9% of biofilm bacteria at higher concentrations than MIC and MBC. These MBIC and MBEC concentrations are much lower than known noxious doses, and the MIC, MBC, and MBIC values were even lower than clinical concentrations.
Bacteria ; Biofilms ; Erythrosine ; Methods ; Microbial Sensitivity Tests ; Plankton

BACKGROUND: Timely intervention in the treatment of bloodstream infection is important for prescription of appropriate antimicrobials. With prompt determination of the antimicrobial susceptibility of a causative agent, rapid antimicrobial susceptibility test (AST) can help select the appropriate antimicrobial therapy. This clinical study is for evaluation of the clinical performance of the QMAC-dRAST for rapid AST directly from positive blood culture (PBC)s with Gram-positive cocci. METHODS: A total of 115 PBC samples with Gram-positive organisms (76 Staphylococcus spp. and 39 Enterococcus spp.) were evaluated by the QMAC-dRAST system, and their pure culture isolates were evaluated by the MicroScan WalkAway (Beckman Coulter, USA) as the comparative AST system. Thirteen antimicrobial agents were included, and the agreement and discrepancy rates of the QMAC-dRAST system (Quantamatrix Inc., Republic of Korea) compared to the MicroScan WalkAway were calculated. To resolve discrepancies, the broth microdilution method was performed. RESULTS: The QMAC-dRAST system exhibited a categorical agreement rate of 94.9% (1,126/1,187) and an essential agreement rate of 98.3% (1,167/1,187). The QMAC-dRAST system yielded very major (false-susceptible) errors at 1.0% (5/485), major (false-resistant) errors at 1.3% (9/693), and minor errors at 4.0% (47/1,187) compared to the MicroScan WalkAway. The QMAC-dRAST system significantly eliminated 30 hours of total turnaround time by combination of direct inoculation of PBC and an image-based approach. CONCLUSION: The results of the QMAC-dRAST system were highly accurate. Thereby, the QMAC-dRAST may provide essential information to accelerate therapeutic decisions for earlier and adequate antibiotic treatment and patient management in clinical settings.
Anti-Infective Agents ; Bacteremia ; Bioengineering ; Clinical Study ; Drug Resistance, Microbial ; Enterococcus ; Gram-Positive Cocci* ; Humans ; Methods ; Microbial Sensitivity Tests ; Prescriptions ; Staphylococcus

This study aimed to evaluate the antimicrobial effects of Acanthopanax sessiliflorum fruit (ASF; Ogaza) extracts on Streptococcus mutans and Streptococcus sobrinus, which are agents that cause dental caries, and on Streptococcus mitis and Streptococcus salivarius, the microbial flora of the oral cavity. The ASF extracts obtained using 70% ethanol were fractionated in the order of ethyl acetate and n-Butanol, concentrated under reduced pressure, and lyophilized to give powdery solvent extracts. The antimicrobial activity of ASF extracts from each solvent was examined using the disk diffusion method. As a result, only those extracts obtained using an ethyl acetate solvent showed antimicrobial activity. These extracts were selected, and the minimum inhibitory concentration was measured by disk diffusion method at various extract concentrations. Results showed a minimum inhibitory concentration of 32 mg/ml. The viable cell count was measured to confirm the minimum bactericidal concentration. Results showed a minimum bactericidal concentration of 64 mg/ml. In the cytotoxicity test using normal human dermal fibroblast cells, the absorbance value of the test group was similar to that of the control group at 0.64, 1.28, and 6.4 mg/ml. The bacteria and their colonies were examined using a scanning electron microscope. Boundaries between the antimicrobial activity region and non-antimicrobial activity region were observed around the paper disk, which was immersed in the extract with 32 mg/ml concentration. Bacterial colonization was not observed in the area with antimicrobial activity. This finding suggests that ASF extracts can inhibit the growth of some microorganisms in the oral cavity, in addition to the effects of these extracts known to date. In particular, ASF extracts may be used as a preparation for preventing dental caries by adding the extract to the toothpaste or oral mouthwash.
1-Butanol ; Bacteria* ; Cell Count ; Colon ; Dental Caries ; Diffusion ; Eleutherococcus* ; Ethanol ; Fibroblasts ; Fruit* ; Humans ; Methods ; Microbial Sensitivity Tests ; Mouth ; Streptococcus ; Streptococcus mitis ; Streptococcus mutans ; Streptococcus sobrinus ; Toothpastes

BACKGROUND: From January 2014 to December 2015, 69 clones of Enterobacter cloacae showing multidrug resistance to six classes of antimicrobial agents were collected from two medical centers in Korea. METHODS: Minimum inhibitory concentrations were determined using the E-test method, and 17 genes were detected using polymerase chain reaction (PCR). The epidemiological relatedness of the strains was identified using repetitive element sequence-based PCR and multilocus sequence typing. RESULTS: The 69 E. cloacae clones produced extended spectrum β lactamase (ESBL) and AmpC and showed multidrug resistance to cefotaxime, ceftazidime, and aztreonam. We identified the following sequence types: ST56 of type VI for ESBL SHV (N=12, 17.4%); ST53, ST114, ST113, and ST550 of types I, IV, VI, and VII, respectively, for CTX-M (N=11, 15.9%); and ST668 of type III for the carbapenemase NDM gene (N=1, 1.5%). The AmpC DHA gene (N=2, 2.89%) was confirmed as ST134, although its type was not identified, whereas EBC (MIR/ACT; N=18, 26.1%) was identified as ST53, ST24, ST41, ST114, ST442, ST446, ST484, and ST550 of types V, I, III, IV, VII, and VI, respectively. The formed subclasses were bla CTX-M-3 and bla CTX-M-22 by CTX-M-1, bla CTX-M-9 and bla CTX-M-125 by CTX-M-9, bla DHA-1 by DHA, and bla MIR-7 and bla ACT-15,17,18,25,27,28 by EBC (MIR/ACT). CONCLUSIONS: There were no epidemiological relationships between the gene products and the occurrence of resistance among the strains.
Anti-Infective Agents ; Aztreonam ; Cefotaxime ; Ceftazidime ; Cloaca ; Clone Cells ; Drug Resistance, Multiple* ; Enterobacter cloacae* ; Enterobacter* ; Korea ; Methods ; Microbial Sensitivity Tests ; Multilocus Sequence Typing ; Polymerase Chain Reaction