Microdilution Method
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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.


Antibiotics ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 87
Author(s):  
Karolína Švarcová ◽  
Marcela Pejchalová ◽  
David Šilha

The purpose of this study was to test the in vitro effects of ampicillin, ciprofloxacin, clindamycin, erythromycin, gentamicin, and tetracycline on planktonic cells of Arcobacter-like microorganisms and on their biofilm formation ability. The minimum inhibitory concentrations (MICs) were determined by the microdilution method. Further, biofilm formation ability in the presence of various concentrations of antibiotics was evaluated by a modified Christensen method. Most of the 60 strains exhibited high susceptibility to gentamicin (98.3%), ciprofloxacin (95.0%), and erythromycin (100.0%). High level of resistance was observed to clindamycin and tetracycline with MIC50 and MIC90 in range of 4–32 mg/L and 32–128 mg/L, respectively. Combined resistance to both clindamycin and tetracycline was found in 38.3% of tested strains. In general, higher biofilm formation was observed especially at lower concentrations of antibiotics (0.13–2 mg/L). However, a significant decrease in biofilm formation ability of Pseudarcobacter defluvii LMG 25694 was exhibited with ampicillin and clindamycin at concentrations above 32 or 8 mg/L, respectively. Biofilm formation represents a potential danger of infection and also a risk to human health, in particular due to antimicrobial-resistant strains and the ability to form a biofilm structure at a concentration that is approximately the MIC determined for planktonic cells.


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.


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 >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.


2022 ◽  
Vol 15 (1) ◽  
pp. 55
Author(s):  
Lais Cavalcanti dos Santos Velasco de Souza ◽  
Lucas Martins Alcântara ◽  
Pãmella Antunes de Macêdo-Sales ◽  
Nathália Faria Reis ◽  
Débora Sena de Oliveira ◽  
...  

Recently, the well-known geographically wide distribution of sporotrichosis in Brazil, combined with the difficulties of effective domestic feline treatment, has emphasized the pressing need for new therapeutic alternatives. This work considers a range of synthetic derivatives as potential antifungals against Sporothrix brasiliensis isolated from cats from the hyperendemic Brazilian region. Six S. brasiliensis isolates from the sporotrichotic lesions of itraconazole responsive or non-responsive domestic cats were studied. The minimum inhibitory concentrations (MICs) of three novel hydrazone derivatives and eleven novel quinone derivatives were determined using the broth microdilution method (M38-A2). In silico tests were also used to predict the pharmacological profile and toxicity parameters of these synthetic derivatives. MICs and MFCs ranged from 1 to >128 µg/mL. The ADMET computational analysis failed to detect toxicity while a good pharmacological predictive profile, with parameters similar to itraconazole, was obtained. Three hydrazone derivatives were particularly promising candidates as antifungal agents against itraconazole-resistant S. brasiliensis from the Brazilian hyperendemic region. Since sporotrichosis is a neglected zoonosis currently spreading in Latin America, particularly in Brazil, the present data can contribute to its future control by alternative antifungal drug design against S. brasiliensis, the most virulent and prevalent species of the hyperendemic context.


2021 ◽  
Vol 18 (4) ◽  
pp. 33-40
Author(s):  
D. V. Tapalski ◽  
E. V. Karpova

Objective. To assess the susceptibility of K.pneumoniae and A.baumanii strains isolated from hospitalized COVID-19 patients to antibiotics and their combinations.Materials and methods. The minimum inhibitory concentrations (MICs) of meropenem and colistin were determined for 47 A.baumannii and 51K.pneumoniaestrains isolated from the hospitalized COVID-19 patients by the broth microdilution method. The susceptibility to 11 antibiotic combinations was assessed using the method of multiple combination bactericidal testing.Results. Colistin resistance was detected in 31.9 % of A.baumannii strains (MIC50 — 0.5 mg/l, MIC90 — 16 mg/l) and in 80.4 % of K.pneumoniaestrains (MIC50 — 16 mg/l, MIC90 — 256 mg/l). It has been shown that double antibiotic combinations with the inclusion of colistin exhibit bactericidal or bacteriostatic activity against 76.6–87.2 % of A.baumannii strains. Combinations with the addition of meropenem, colistin and macrolides exhibited bactericidal activity against 78.4–80.4 % of K.pneumoniae strains. Combinations of two carbapenems were not active, the combination of meropenem-colistin had a bactericidal effect only in 13.7 % of K.pneumoniae strains.Conclusion. Widespread colistin resistance was found in carbapenem-resistant K.pneumoniae and A.baumannii strains isolated from the hospitalized COVID-19 patients. The combinations of antibiotics that have a synergistic antibacterial effect in their pharmacokinetic/pharmacodynamic concentrations have been determined.


2021 ◽  
Vol 23 (1) ◽  
pp. 284
Author(s):  
Kamila Korzekwa ◽  
Anna Kędziora ◽  
Bartłomiej Stańczykiewicz ◽  
Gabriela Bugla-Płoskońska ◽  
Dorota Wojnicz

The aim of this study was to assess the beneficial inhibitory effect of silver nanoparticles immobilized on SiO2 or TiO2 on biofilm formation by Pseudomonas aeruginosa—one of the most dangerous pathogens isolated from urine and bronchoalveolar lavage fluid of patients hospitalized in intensive care units. Pure and silver doped nanoparticles of SiO2 and TiO2 were prepared using a novel modified sol-gel method. Ten clinical strains of P. aeruginosa and the reference PAO1 strain were used. The minimal inhibitory concentration (MIC) was determined by the broth microdilution method. The minimal biofilm inhibitory concentration (MBIC) and biofilm formation were assessed by colorimetric assay. Bacterial enumeration was used to assess the viability of bacteria in the biofilm. Silver nanoparticles immobilized on the SiO2 and TiO2 indicated high antibacterial efficacy against P. aeruginosa planktonic and biofilm cultures. TiO2/Ag0 showed a better bactericidal effect than SiO2/Ag0. Our results indicate that the inorganic compounds (SiO2, TiO2) after nanotechnological modification may be successfully used as antibacterial agents against multidrug-resistant P. aeruginosa strains.


2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Nahla Ayoub ◽  
Nadia Badr ◽  
Saeed S Al-Ghamdi ◽  
Arwa Alzahrani ◽  
Rahaf Alsulaimani ◽  
...  

Introduction. Salvadora persica L. (S. persica, Siwak) has been used for many centuries as oral hygiene tools, particularly in Saudi Arabia. This study aimed to assess the effectiveness of S. persica petroleum ether extract (SPE) as an intracanal bactericidal for endodontic treatment against Enterococcus faecalis. Calcium hydroxide Ca(OH)2 gold standard intracanal medicament was used for comparison. Methods. The gas chromatography mass spectrometry (GC/MS) analysis was carried out to identify the components of SPE. First, the consistency of SPE was accomplished according to ANSI/ADA specification no 57. Forty-five single-rooted mandibular premolars were infected with that of E. faecalis suspension. Colony-forming units (CFU) were counted before the medicaments’ application (CFU-1) and after seven days of their applications (CFU-2). Group I: SPE, Group II: positive control Ca(OH)2, and Group III: saline solution negative control. The microdilution method was applied to determine minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of SPE. Results. Thirty-two compounds were identified (89.09%), with main components of benzyl isothiocyanate (BITC) (33.32%) and steroids (34%). CFU before and after using SPE and Ca(OH)2 recorded a statistically significant reduction in bacterial count ( P = 0.006 ) and ( P = 0.01 ), respectively. There was an insignificant difference between CFU after using SPE and Ca(OH)2 ( P = 0.210 ). On the contrary, comparing both medicaments with the negative control saline group resulted in significant differences, ( P = 0.001 ) and ( P = 0.007 ), respectively. Moreover, the equality of minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) of SPE is recorded. Conclusion. This finding could be referred to the high content of bactericidal BITC in synergism with other antimicrobial components, representing 70.71% of SPE. Thus, SPE is a good candidate as an intracanal medicament, which warrants further investigation.


2021 ◽  
Vol 14 (11) ◽  
Author(s):  
Yuhui Wang ◽  
Jie Ma ◽  
Wanxiang Li ◽  
Mi Liu ◽  
Yansheng Ding

Background: In recent years, the widespread use of antibiotics has resulted in increased rates of antibiotic resistance (ABR). Pseudomonas aeruginosa is one of the most important opportunistic pathogens causing hospital-acquired infections. Pseudomonas aeruginosa has continuously increased resistance to commonly used clinical antimicrobial drugs, bringing great difficulties to clinical treatment. Objectives: This retrospective study investigated the epidemiological characteristics of P. aeruginosa and changes in ABR over a 5-year period at a hospital in Shandong Province, China. Methods: Pseudomonas aeruginosa strains were collected from 2015 to 2019. The antimicrobial susceptibility testing employed the Kirby-Bauer disk diffusion method and the broth microdilution method (VITEK-2 compact system), according to the guidelines by the Clinical and Laboratory Standards Institute. Data were analyzed using WHONET 5.6 and SPSS V. 21.0 software. Results: A total of 3,324 P. aeruginosa strains were isolated from clinical specimens (604, 631, 700, 595, and 794 strains from 2015 to 2019, respectively). The highest P. aeruginosa detection rates were from respiratory tract specimens (72.54%). The highest resistance was seen in aztreonam, followed by ciprofloxacin, levofloxacin, and imipenem. The isolation rates for carbapenem-resistant P. aeruginosa (CRPA) and multidrug-resistant P. aeruginosa (MDRPA) ranged from 15.21 - 18.38% and 17.31 - 27.31%, respectively. Also, the isolation rates for extensively drug-resistant P. aeruginosa (XDRPA) ranged from 1.86 - 3.52%. Conclusions: The main sources of the P. aeruginosa isolates were older adult patients with chronic respiratory diseases. The isolation rates for CRPA, MDRPA, and XDRPA strains decreased over the 5-year period. However, the drug resistance situation remains a serious concern. Hence, continued infection control and antimicrobial stewardship and basic and clinical research on bacterial resistance are essential.


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