Broth Microdilution
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2022 ◽  
Vol 12 ◽  
Author(s):  
Menglan Zhou ◽  
Ziran Wang ◽  
Li Zhang ◽  
Timothy Kudinha ◽  
Haoran An ◽  
...  

Background:Streptococcus pneumoniae is an important human pathogen that can cause severe invasive pneumococcal diseases (IPDs). The aim of this multicenter study was to investigate the serotype and sequence type (ST) distribution, antimicrobial susceptibility, and virulence of S. pneumoniae strains causing IPD in China.Methods: A total of 300 invasive S. pneumoniae isolates were included in this study. The serotype, ST, and antimicrobial susceptibility of the strains, were determined by the Quellung reaction, multi-locus sequence typing (MLST) and broth microdilution method, respectively. The virulence level of the strains in the most prevalent serotypes was evaluated by a mouse sepsis model, and the expression level of well-known virulence genes was measured by RT-PCR.Results: The most common serotypes in this study were 23F, 19A, 19F, 3, and 14. The serotype coverages of PCV7, PCV10, PCV13, and PPV23 vaccines on the strain collection were 42.3, 45.3, 73.3 and 79.3%, respectively. The most common STs were ST320, ST81, ST271, ST876, and ST3173. All strains were susceptible to ertapenem, levofloxacin, moxifloxacin, linezolid, and vancomycin, but a very high proportion (>95%) was resistant to macrolides and clindamycin. Based on the oral, meningitis and non-meningitis breakpoints, penicillin non-susceptible Streptococcus pneumoniae (PNSP) accounted for 67.7, 67.7 and 4.3% of the isolates, respectively. Serotype 3 strains were characterized by high virulence levels and low antimicrobial-resistance rates, while strains of serotypes 23F, 19F, 19A, and 14, exhibited low virulence and high resistance rates to antibiotics. Capsular polysaccharide and non-capsular virulence factors were collectively responsible for the virulence diversity of S. pneumoniae strains.Conclusion: Our study provides a comprehensive insight into the epidemiology and virulence diversity of S. pneumoniae strains causing IPD in China.


Author(s):  
Renru Han ◽  
Xuelin Yang ◽  
Yang Yang ◽  
Yan Guo ◽  
Dandan Yin ◽  
...  

Multidrug-resistant Gram-negative bacteria, especially for extended-spectrum β-lactamases-producing and carbapenemase-producing Enterobacterales , are disseminating rapidly around the world. Treatment options for these infections are limited, which prompt the development of novel or combinational therapies to combat the infections caused by multidrug-resistant pathogens.


Nanomedicine ◽  
2022 ◽  
Author(s):  
Jiří Trousil ◽  
Jana Matějková ◽  
You-Shan Dai ◽  
Tomáš Urbánek ◽  
Miroslav Šlouf ◽  
...  

Background: Antimicrobial submicrometer particles are being studied as promising interventions against a wide range of skin conditions, such as fungal or bacterial infections. Aims: To submicronize chloroxine, the crystalline compound 5,7-dichloro-8-hydroxyquinoline, by nanoprecipitation and characterize the resulting assemblies. Methods: The chloroxine particles were stabilized by a nonionic surfactant and were studied by a broth microdilution assay against 20 medically important bacteria and fungi. The intervention was studied using a murine model of skin irritation. Results & conclusions: Chloroxine nanoparticles with a diameter of 600–800 nm exhibit good tolerability in terms of skin irritation in vivo and good antimicrobial activity. Thus, the fabricated formulation shows great promise for interventions for both cutaneous infection control and prophylaxis.


Author(s):  
Soudabeh Alizadeh Matboo ◽  
Shahram Nazari ◽  
Ali Niapour ◽  
Mehdi Vosoughi Niri ◽  
Esrafil Asgari ◽  
...  

Abstract This study investigated the bacterial removal using TiO2 nanoparticles (NPs) modified with poly-amidoamine dendrimer macromolecule (PAMAM, G3). The PAMAM G3/TiO2 (nanohybrid) was used to specify antibacterial properties via broth microdilution (MBC-Minimum Bactericidal Concentration and MIC-Minimum Inhibitory Concentration- determination), paper disc diffusion, and surface plate count methods. The nanohybrid was characterized via the different techniques. The effects of different factors including initial bacteria count, run time, solution pH, and the nanohybrid concentration were studied. The nanohybrid cytotoxicity was studied on AGS and MKN45 cells line by MTT assay. It was revealed that the nanohybrid was effective in intercepting both bacterial strains growth. The MIC value for S. aureus and E. coli were determined to be 4 and 2 μg/mL, respectively. The MBC value for both strains were calculated to be 32 μg/mL. The results showed removal efficiency of 100% for S. aureus and E. coli bacteria in optimum situation. The decrease in cell viability in the dosage of 32 μg/mL after 72 h treatment for AGS and MKN45 cells line were shown to be 6.2 and 4.6%, respectively. The nanohybrid was able to decrease the S. aureus and E. coli count in solution, which meets the drinking water criterions aligned with WHO guidelines.


2022 ◽  
Vol 12 ◽  
Author(s):  
Dokyun Kim ◽  
Eun-Jeong Yoon ◽  
Jun Sung Hong ◽  
Min Hyuk Choi ◽  
Hyun Soo Kim ◽  
...  

To monitor national antimicrobial resistance (AMR), the Korea Global AMR Surveillance System (Kor-GLASS) was established. This study analyzed bloodstream infection (BSI) cases from Kor-GLASS phase I from January 2017 to December 2019. Nine non-duplicated Kor-GLASS target pathogens, including Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter spp., and Salmonella spp., were isolated from blood specimens from eight sentinel hospitals. Antimicrobial susceptibility testing, AMR genotyping, and strain typing were carried out. Among the 20,041 BSI cases, 15,171 cases were caused by one of the target pathogens, and 12,578 blood isolates were collected for the study. Half (1,059/2,134) of S. aureus isolates were resistant to cefoxitin, and 38.1% (333/873) of E. faecium isolates were resistant to vancomycin. Beta-lactamase-non-producing ampicillin-resistant and penicillin-resistant E. faecalis isolates by disk diffusion method were identified, but the isolates were confirmed as ampicillin-susceptible by broth microdilution method. Among E. coli, an increasing number of isolates carried the blaCTX–M–27 gene, and the ertapenem resistance in 1.4% (30/2,110) of K. pneumoniae isolates was mostly (23/30) conferred by K. pneumoniae carbapenemases. A quarter (108/488) of P. aeruginosa isolates were resistant to meropenem, and 30.5% (33/108) of those carried acquired carbapenemase genes. Over 90% (542/599) of A. baumannii isolates were imipenem-resistant, and all except one harbored the blaOXA–23 gene. Kor-GLASS provided comprehensive AMR surveillance data, and the defined molecular mechanisms of resistance helped us to better understand AMR epidemiology. Comparative analysis with other GLASS-enrolled countries is possible owing to the harmonized system provided by GLASS.


2022 ◽  
Author(s):  
Sônia Maria Rolim Rosa Lima ◽  
Maria Thereza Gamberini ◽  
Domingos Sávio Rodrigues ◽  
Pedro Ismael Silva Junior ◽  
Kátia Andrea de Menezes Torres

Abstract Maternal colonization by Group B Streptococcus during pregnancy increases the risk of neonatal infection due to vertical transmission from mother to fetus before or during labor. The aims of this study were to evaluate the antimicrobial activity of SP80 (obtained from RGE) and its synergism associated with the antibiotic against strains of Streptococcus agalactiae. Biomonitoring of SP80 disclosed antimicrobial activity only in fractions F18, F19, F20 and F42. The broth microdilution was used to determine the antimicrobial activity of SP80 and fractions from SP80 and to establish the MIC of SP80 (2.40 mg/mL). By using the disk diffusion method, fifty-five clinical isolates of S. agalactiae and 1 ATCC were tested against the association of SP80 with antibiotic penicillin G and ampicillin, respectively, for synergistic assessment. The association of SP80 with penicillin G showed that the mean of the inhibition halos decreased, but it was not significant, with p<0.07. In contrast, the association of SP80 with ampicillin caused the mean inhibition halos to increase with a p<0.001, a significant result. SP80 has antimicrobial activity against S. agalactiae Gram-positive bacteria, and the association with the antibiotic ampicillin showed a synergistic effect, which did not occur when in association with penicillin G.


2022 ◽  
Vol 14 (11) ◽  
Author(s):  
Mehmet Akif Gun ◽  
Yeliz Tanriverdi Cayci ◽  
Belma Durupinar ◽  
Ahmet Yilmaz Coban

Background: The quick diagnosis and early initiation of antibiotic therapy in bacteria-induced infections is of paramount importance. Accordingly, the rapid identification of the causative agent, the short-term results of antibiotic sensitivity, the selection and use of right antibiotics for treatment further highlights the significance of this issue. Objectives: This study aimed to develop a new susceptibility testing method to provide rapid results in Escherichia coli clinical isolates and report the antibiotic susceptibility test results to clinicians in a short period. Methods: In the study, one hundred and ten E. coli clinical isolates were tested. In this regard, antibiotics recommended by the "Clinical and Laboratory Standards Institute (CLSI)" for testing the sensitivity of E. coli isolates, including amoxicillin-clavulanate, cefixime, ceftriaxone, ertapenem, ciprofloxacin, gentamicin, trimethoprim-sulfamethoxazole, and nitrofurantoin were tested. For quality control, E. coli ATCC25922, E. coli ATCC35218, Staphylococcus aureus ATCC29213, and E. coli 13846NTCC strains were used. The broth microdilution method recommended by CLSI was used as the reference method. Minimum inhibitory concentration values were determined, and antimicrobial susceptibilities were then determined according to the “European Committee on Antimicrobial Susceptibility Testing (EUCAST)” criteria. In the next phase, the results of the resazurin microplate method (RMM) were compared. Results: The comparison of the RMM developed in the present study with the reference method revealed that the calculated essential agreement ratios for eight antibiotics varied from 82.72 to 100%, and the categorical agreement values ranged from 95.45 to100%. Conclusions: According to the findings, the RMM results were highly in agreement with the results of the reference method. RMM allows the detection of antibiotic susceptibility quickly (e.g., within 5 hours) as such it is preferred, especially for laboratories with limited facilities. However, further multi-center studies are recommended to use this method in routine laboratories.


2022 ◽  
Vol 12 ◽  
Author(s):  
Lulu Zhang ◽  
Weifeng Yang ◽  
Yajun Chu ◽  
Bo Wen ◽  
Yungchi Cheng ◽  
...  

Methicillin-resistant Staphylococcus aureus (MRSA) is a superbacterium, and when it forms biofilms, it is difficult to treat even with the first-line of antibiotic linezolid (LNZ). Reyanning mixture (RYN), a compound-based Chinese medicine formula, has been found to have inhibitory effects on biofilms. This study aims to explore the synergistic inhibitory effect and corresponding mechanisms of their (LNZ&amp;RYN) combination on the planktonic as well as biofilm cells of MRSA. Broth microdilution and chessboard methods were employed for the determination of minimum inhibitory concentrations (MICs) and synergistic concentration of LNZ&amp;RYN, respectively. The effect of the combined medication on biofilm and mature biofilm of MRSA were observed by biofilm morphology and permeability experiments, respectively. To unveil the molecular mechanism of action of the synergistic combination of LNZ and RYN, RT-PCR based biofilm-related gene expression analysis and ultra-high pressure liquid chromatography-time-of-flight mass spectrometry based endogenous metabonomic analysis were deployed. The results indicated that 1/16RYN as the best combined dose reduced LNZ (4 μg/ml) to 2 μg/ml. The combined treatment inhibited living MRSA before and after biofilm formation, removed the residual structure of dead bacteria in MRSA biofilms and affected the shape and size of bacteria, resulting in the improvement of biofilm permeability. The mechanism was that biofilm-related genes such as agrC, atlA, and sarA, as well as amino acid uptake associated with the metabolism of 3-dehydrocarnitine, kynurenine, L-leucine, L-lysine and sebacic acid were inhibited. This study provides evidence for the treatment of MRSA and its biofilms with LNZ combined with RYN.


Author(s):  
Siquan Shen ◽  
Xiangning Huang ◽  
Qingyu Shi ◽  
Yan Guo ◽  
Yang Yang ◽  
...  

Providencia rettgeri is a nosocomial pathogen associated with urinary tract infections related to hospital-acquired Infections. In recent years, P. rettgeri clinical strains producing New Delhi Metallo-β-lactamase (NDM) and other β-lactamase which reduce the efficiency of antimicrobial therapy have been reported. However, there are few reports of P. rettgeri co-producing two metallo-β-lactamases in one isolate. Here, we first reported a P. rettgeri strain (P138) co-harboring blaNDM-1, blaVIM-1, and blaOXA-10. The specie were identified using MALDI-TOF MS. The results of antimicrobial susceptibility testing by broth microdilution method indicated that P. rettgeri P138 was resistant to meropenem (MIC = 64μg/ml), imipenem (MIC = 64μg/ml), and aztreonam (MIC = 32μg/ml). Conjugation experiments revealed that the blaNDM-1-carrying plasmid was transferrable. The carbapenemase genes were detected using PCR and confirmed by PCR-based sequencing. The complete genomic sequence of the P. rettgeri was identified using Illumina (Illumina, San Diego, CA, USA) short-read sequencing (150bp paired-end reads), and many common resistance genes had been identified, including blaNDM-1, blaVIM-1, blaOXA-10, aac(6’)-Il, aadA5, ant(2’’)-Ia, aadA1, aac(6’)-Ib3, aadA1, aph(3’)-Ia, aac(6’)-Ib-cr, qnrD1, qnrA1, and catA2. The blaNDM-1 gene was characterized by the following structure: IS110–TnpA–IntI1–aadB–IS91–GroEL–GroES–DsbD–PAI–ble–blaNDM-1–IS91–QnrS1–IS110. Blast comparison revealed that the blaNDM-1 gene structure shared &gt;99% similarity with plasmid p5_SCLZS62 (99% nucleotide identity and query coverage). In summary, we isolated a P. rettgeri strain coproducing blaNDM-1, blaVIM-1, and blaOXA-10. To the best of our acknowledge, this was first reported in the world. The occurrence of the strain needs to be closely monitored.


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