Essential Agreement Research Articles

520. Development of Cefepime-taniborbactam MIC Antimicrobial Susceptibility Test (AST) for Enterobacterales and Pseudomonas aeruginosa on MicroScan Dried Gram-negative MIC Panels

Abstract Background Automated AST devices are critical to inform appropriate care of patients with bacterial infections. Cefepime-taniborbactam is an investigational agent under development to treat infections due to multidrug-resistant bacteria, including carbapenem-resistant Enterobacterales and Pseudomonas aeruginosa. Development of a cefepime-taniborbactam antimicrobial susceptibility test was completed for the MicroScan Dried Gram-negative MIC (MSDGN) Panel when compared to CLSI broth microdilution reference panels. Methods Development was conducted by comparing MICs obtained using the MSDGN panel to MICs using a CLSI broth microdilution reference panel. The concentration range of cefepime-taniborbactam evaluated was 0.12/4-64/4 µg/mL. A total of 1376 isolates (1256 Enterobacterales and 120 P. aeruginosa) were tested at 16, 18, and 20 hours incubation times (for visual read and autoSCAN-4). A total of 917 isolates (837 Enterobacterales and 80 P. aeruginosa) were tested at 16 and 18 hours incubation time for the WalkAway System. All isolates were tested using turbidity and Prompt® methods of inoculation. MSDGN panels were incubated at 35 ± 1°C. Reference panels, prepared according to ISO methodology, were inoculated using the turbidity inoculation method. All frozen reference panels were incubated at 35 ± 2°C and read visually. Rates of essential agreement (MicroScan MIC within ± 1 doubling dilution of reference MIC) were determined for each combination of inoculation method and read method. Results Essential agreement rates ranged from 93.0% to 98.8% for all isolates tested during development. Essential agreement rates for each inoculation and read method are presented in Table 1. Table 1- Results Conclusion Cefepime-taniborbactam MIC results obtained with the MSDGN panel correlated well with MIC results obtained using reference panels. Rates of essential agreement were ≥93.0% for all inoculation and read methods. These development data support the continued evaluation of MSDGN panel with cefepime-taniborbactam in a multicenter trial. Pending submission and clearance by the United States Food and Drug Administration; not yet available for in vitro diagnostic use in the US. For Investigational Use Only. The performance characteristics of this product have not been established. Disclosures Enrique J. Fernandez, BS, Beckman Coulter, Inc.: Employee of Beckman Coulter Robert L. Williams, PhD, Beckman Coulter, Inc.: Employee of Beckman Coulter Vasna Carr, BS, Beckman Coulter, Inc.: Employee of Beckman Coulter Vasna Carr, BS, Beckman Coulter, Inc.: Employee of Beckman Coulter Miller Renae, BS, Beckman Coulter, Inc.: Employee of Beckman Coulter.

Evaluation of three commercial methods of susceptibility testing for ceftolozane/tazobactam against carbapenem-resistant Pseudomonas aeruginosa

IntroductionCeftolozane/tazobactam has shown excellent activity against Pseudomonas aeruginosa, but this drug is not always included in commercial panels. The aim of the study was to evaluate the performance of 2 gradient strips (BioMérieux and Liofilchem) and a commercial microdilution panel (Sensititre, EURGNCOL panel) using this combination against carbapenem-resistant P. aeruginosa isolates. MethodsThree commercial methods were tested with 41 metallo-beta-lactamase-producing and 59 non-carbapenemase-producing P. aeruginosa isolates. Broth microdilution was used as reference. ResultsAll carbapenemase-producing isolates and only one non-producing isolate were resistant to this antibiotic. Both essential agreement and bias were outside the acceptance intervals since MIC values were higher than reference values for all three methods. The Kappa index indicated poor or weak agreement. Changes in clinical categories were observed in 3 isolates. ConclusionsThe three methods yielded poor agreement with the reference. Despite the differences in MIC values, fewer than 3% involved category changes.

Detection of colistin resistance via four methods in multidrug-resistant Gram-negative rods isolated from blood cultures.

The broth microdilution (BMD) method recommended for the detection of colistin resistance is labor-intensive, time-consuming, and difficult to apply in routine laboratories. Thus, various methods, such as disk elution, commercial microdilution, and rapid polymyxin-NP tests have been developed for the detection of colistin resistance. In this study, a total of 102 multi-resistant Gram-negative bacteria isolated from blood cultures were evaluated by four different methods for the detection of colistin resistance, and compared with the reference method. For the detection of the compatibility of these methods with the reference method, categorical and essential agreements, very major, major, and minor error rates were determined. Colistin-tigecycline and colistin-meropenem combinations were investigated in colistin-resistant isolates. Of the isolates, 15 (15%) [K. pneumoniae (n = 12), A. baumannii (n = 2), E. coli (n = 1)] were resistant to colistin with reference BMD method. MIC50 and MIC90 values of all isolates were ≤ 0.25 μg/mL and 16 μg/mL, respectively. The categorical agreement rates were 100% for commercial microdilution, disk elution, and RPNP test. The essential agreement rates of commercial microdilution, disk elution, and broth macrodilution were 78.4%, 86.3%, and 100%, respectively. Although there were no major errors in these methods, the macrodilution (12%) and commercial microdilution (20.6%) methods showed the most minor errors. Colistin-meropenem combination showed a 100% synergistic effect, but the colistin-tigecycline combination showed an 80% synergistic effect and 20% indifference effect. Disk elution and RPNP tests are suitable for routine use because they are the most efficient, easiest, low-cost, and good performance tests in detecting colistin resistance.

Evaluation of Dilution Susceptibility Testing Methods for Aztreonam in Combination with Avibactam against Enterobacterales.

As multidrug and pan-resistance among Enterobacterales continue to increase, there is an urgent need for more therapeutic options to treat these infections. New β-lactam and β-lactam inhibitor (BLI) combinations have a broad spectrum of activity, but those currently approved do not provide coverage against isolates harboring metallo-β-lactamases (MBL). Aztreonam (ATM) and avibactam (AVI) in combination (ATM/AVI; AVI at 4 μg/mL fixed concentration) provides a similarly broad range of activity while maintaining activity against MBL-producing isolates. The in vitro susceptibility testing of ATM/AVI by standard methods was evaluated during development. This study investigated the impact of nonstandard testing conditions on the activity of ATM/AVI as observed during broth microdilution testing as well as the equivalency between agar dilution and broth microdilution MIC values when testing a diverse panel of Enterobacterales (N = 201). Nonstandard test conditions evaluated included inoculum density, atmosphere of incubation, media pH, varied medium cation concentrations, incubation time, varied serum concentrations, testing in pooled urine instead of media, addition of blood to the media, and the presence of surfactant. Generally, apart from low pH and high inoculum density, nonstandard testing parameters did not affect ATM/AVI broth microdilution MIC values. Correlation of MIC values obtained by agar dilution and broth microdilution resulted in an essential agreement of 97.0% for all tested Enterobacterales. Variation of standard testing conditions had little impact on broth microdilution MIC values for ATM/AVI. The correlation between broth microdilution and agar dilution MICs suggests both methods are reliable for determination of ATM/AVI MIC values. IMPORTANCE Increasing antibiotic resistance and emergence of pan-resistant isolates threaten the ability to control infections and to provide many other medical interventions such as surgery and chemotherapy, among others. New therapies are required to control emerging resistance mechanisms, including the increase in metallo-β-lactamases. Some new antibiotic combinations provide coverage against highly resistant isolates but are unable to target organisms that produce metallo-β-lactamases. Aztreonam in combination with avibactam provides a broad spectrum of activity against highly resistant isolates that also targets metallo-β-lactamase-producing organisms. An important part of drug development is the ability for clinical labs to determine the susceptibility of isolates to the antimicrobial. This manuscript investigates the in vitro susceptibility testing of aztreonam/avibactam with nonstandard testing conditions and a correlation study between broth microdilution and agar dilution against clinical isolates encoding a variety of resistance mechanisms. Overall, aztreonam/avibactam was generally unaffected by changes in testing conditions and showed strong agar/broth correlation.

The seemingly ordinary complexity of daily life

Abstract The author is in essential agreement with Tallis, that when we only deploy one mode of interpretation, ie the scientific mode, we lose the fundamental realities of human experience, including the experience of free will, on which, ironically, scientific practice depends. Tallis’s philosophical stance is compared to that of Owen Barfield and his work on free will is placed within the context of his other books. A sense of wonder is common to all of them.

Genomic analysis of Fosfomycin resistance in multi-drug resistant uropathogens and comparison of in-vitro susceptibility methods uropathogens.

Urinary tract infection is one of the most common bacterial infections causing high morbidity and mortality. The alarming rise of multidrug-resistant uropathogens worldwide forced the clinician to rethink the old drugs like Fosfomycin for its therapeutic management. Our objective was to compare agar dilution, disc diffusion and E-test method for antimicrobial susceptibility testing of Fosfomycin against different drug-resistant uropathogens. Consecutive 181 uropathogens were tested for Fosfomycin susceptibility using agar dilution, disc diffusion and E-test. Results were interpreted using Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints. Whole genome sequencing analysis was done on the 4 XDR/PDR Fosfomycin resistant Klebsiella pneumoniae isolates. Escherichia coli was found as the most common (62.4%) uropathogen followed by Klebsiella pneumoniae (21%). Considering agar dilution as the gold standard, 6.1% of isolates were resistant to Fosfomycin. Following CLSI breakpoints, the susceptibility of Escherichia coli, Klebsiella pneumoniae, other Enterobacterales and Pseudomonas aeruginosa were 92.9%, 92.1%, 100%, 100%; whereas using EUCAST breakpoints the susceptibility rates were 85.7%, 86.9%, 92.9%, and 100%, respectively. The essential agreement, categorical agreement, major error, and very major error for E-test/disc diffusion for all the organisms were 91.2%/Not Applicable, 95%/93.9%, 1.8%/4.7%, 9.1%/9.1%, respectively. Whole-genome sequencing showed mutation UhpT gene as well as the presence of plasmid-mediated fosA5 or fosA6 genes conferring Fosfomycin resistance. This result supports very low resistance of Enterobacterales against Fosfomycin; hence should be considered a valuable option to treat multidrug-resistant uropathogens. Disc diffusion was observed to be a convenient method for Fosfomycin susceptibility testing compared to agar dilution.

A review of European and international phthalates regulation: focus on daily use products

Abstract Background Phthalates are known endocrine disruptors used in a wide range of industrial and household products. With globalization and interdependency of the supply chain, the control of toxic substances in daily use products has become more challenging. Many countries have implemented laws and policies to limit their use, although these regulations are neither unified nor seem adequate, as studies suggest that more vulnerable populations (children, pregnant women) are exposed to phthalates that should be restricted. Methods For seven of the most used phthalates - bis(2-Ethylhexyl) phthalate (DEHP), Butylbenzyl phthalate (BBP), Dibutyl phthalate (DBP), Diisobutyl phthalate (DIBP), diisononyl phthalate (DINP), diisodecyl phthalate (DIDP), Di-n-octyl phthalate (DNOP) - we performed an online research on institutional sites and dedicated Agencies of the three largest world economies (European Union (EU), United States of America (USA), China) to analyze their uses and bans, focusing on Food Contacts Materials (FCM), cosmetics, toys and childcare articles. Results In the EU area DEHP, BBP, DBP and DIBP are not allowed in toys and childcare articles above 0,1% by weight. All the seven phthalates are also severely restricted in FCM, and mostly banned as cosmetic components. In the USA, there is no formal prohibition to their use in cosmetics but phthalates are mostly limited in FCM. In China, the limit for DBP, BBP, DEHP, DNOP, DINP, DIDP in plastic toys is 0,1% of the material composition; regarding cosmetics DEHP, BBP and DBP are prohibited. Conclusions We found substantial differences in the international legislation. Though there is essential agreement on toys and childcare articles legislation, there are many discrepancies about FCM and cosmetics. Further research is needed to compare the regulation with data about concentrations of these ubiquitous elements, to underline the real exposure and risk in different populations and to improve knowledge and safety on this matter. Key messages • Phthalates, known endocrine disruptors, in daily use products are a matter of concern. • Coordinated international laws to prevent exposure, especially in vulnerable populations, are needed.

Direct MALDI-TOF MS and Antimicrobial Susceptibility Testing of Positive Blood Cultures Using the FASTTM System and FAST-PBC Prep Cartridges-Performance Evaluation in a Clinical Microbiology Laboratory Serving High-Risk Patients.

Bloodstream infections are a leading cause of morbidity and mortality. The rapid diagnostic testing of positive blood cultures (PBCs) shortens times to effective therapy and the de-escalation of broad-spectrum empiric therapy. This is the first study examining the Qvella FASTTM System for the rapid (~20 min) purification of microorganisms directly from PBCs using BacT/Alert® FA/FAN bottles in the bioMérieux Virtuo instrument. We compared the performance of the FASTTM System Liquid ColonyTM (LC), for immediate downstream ID and phenotypic AST, to standard workflow involving colonies obtained by overnight subculture. The LC yielded a concordant species ID by VITEK MS in 121/138 (87.7%) samples, identifying 32 different Gram-positive and Gram-negative species with 3/123 (2.6%) discordances. Compared to standard workflow, direct AST of the LC using VITEK® 2 yielded 98.4% categorical agreement and 98.0% essential agreement. Very major error, major error, and minor error rates were 1.0%, 0.0%, and 1.8%, respectively, for Gram-negative organisms; and 1.9%, 0.2%, and 1.2%, respectively, for Gram-positive organisms. The median times from positive blood culture flag to results by FASTTM System for ID and AST were 7.8 h and 15.7 h, respectively, versus 22.4 h and 36.6 h for standard workflow, respectively. In conclusion, the FASTTM System provides reliable results for direct ID and AST from PBCs with significantly decreased turnaround times.

Evaluation of Agar Dilution Method in Susceptibility Testing of Polymyxins for Enterobacteriaceae and Non-Fermentative Rods: Advantages Compared to Broth Microdilution and Broth Macrodilution.

An accurate and reliable susceptibility testing method for polymyxins is urgently needed not only for the clinical laboratory but also for new polymyxin-like lipopeptide development. Reference broth microdilution (rBMD), which was the recommended method by CLSI-EUCAST in clinics, has been proven not to be ideal, while the agar dilution (AD) method that was widely used in new antibiotics discovery has been neglected. In the present study, the AD method was compared with rBMD and broth macrodilution (BMAD) in susceptibility testing of polymyxin B and colistin against >200 Gram-negative isolates. AD showed strong agreement with BMAD for colistin (except for Klebsiella aerogenes and Pseudomonas aeruginosa); however, its performance was poor for polymyxin B or compared to rBMD. MICs of AD method were not affected when different types of Petri dishes were used, while glass-bottom microtiter plates could lower the MIC of polymyxins 2–8 times compared to tissue-culture-treated polystyrene plates when using rBMD, which demonstrated that tissue-culture-treated plates were not suitable. It was then validated with non-tissue-culture-treated plates. The culture volume was another influencing factor of accuracy for rBMD, and 200 μL seemed to be the most suitable volume for MIC detection of polymyxins. Additionally, no lack of growth phenomenon (skipped well) was observed for AD when it frequently occurred for both BMAD and rBMD. As for strains carrying mcr-1 gene, 100% of AD results were in essential agreement (EA) and categorical agreement (CA) with both rBMD and BMAD. Overall, rBMD is convenient and widely accepted for susceptibility testing of polymyxins. Although it may be too early to say that AD is superior compared to rBMD and BMAD, it did show some advantages in repeatability and anti-interference ability.

Evaluation of the Sensititre YeastOne and Etest in Comparison with CLSI M38-A2 for Antifungal Susceptibility Testing of Three Azoles, Amphotericin B, Caspofungin, and Anidulafungin, against Aspergillusfumigatus and Other Species, Using New Clinical Breakpoints and Epidemiological Cutoff Values.

Aspergillosis is an invasive fungal disease associated with high mortality. Antifungal susceptibility testing (AFST) is receiving increasing consideration for managing patients, as well as for surveilling emerging drug resistance, despite having time-consuming and technically complex reference methodologies. The Sensititre YeastOne (SYO) and Etest methods are widely utilized for yeasts but have not been extensively evaluated for Aspergillus isolates. We obtained Posaconazole (POS), Voriconazole (VCZ), Itraconazole (ITC), Amphotericin B (AMB), Caspofungin (CAS), and Anidulafungin (AND) minimum inhibitory concentrations (MICs) for both the Etest (n = 330) and SYO (n = 339) methods for 106 sequenced clinical strains. For 84 A. fumigatus, we analyzed the performance of both commercial methods in comparison with the CLSI-AFST, using available cutoff values. An excellent correlation could be demonstrated for Etest-AMB and Etest-VCZ (p < 0.01). SYO-MICs of AMB, VCZ, and POS resulted in excellent essential agreement (>93%), and >80% for AMB, VCZ, and ITC Etest-MICs. High categoric agreement was found for AMB, ITC, and CAS Etest-MICs (>85%) and AMB SYO-MICs (>90%). The considerable number of major/very major errors found using Etest and SYO, possibly related to the proposed cutoffs and associated with the less time-consuming processes, support the need for the improvement of commercial methods for Aspergillus strains.

Mutation detection and minimum inhibitory concentration determination against linezolid and clofazimine in confirmed XDR-TB clinical isolates

BackgroundThe emergence of multidrug-resistant tuberculosis (MDR-TB) has complicated the situation due to the decline in potency of second-line anti-tubercular drugs. This limits the treatment option for extensively drug-resistant tuberculosis (XDR-TB). The aim of this study was to determine and compare the minimum inhibitory concentration (MIC) by agar dilution and resazurin microtiter assay (REMA) along with the detection of mutations against linezolid and clofazimine in confirmed XDR-TB clinical isolates.ResultsA total of 169 isolates were found positive for Mycobacterium tuberculosis complex (MTBC). The MIC was determined by agar dilution and REMA methods. The isolates which showed non-susceptibility were further subjected to mutation detection by targeting rplC gene (linezolid) and Rv0678 gene (clofazimine). The MIC for linezolid ranged from 0.125 µg/ml to > 2 µg/ml and for clofazimine from 0.25 µg/ml to > 4 µg/ml. The MIC50 and MIC90 for linezolid were 0.5 µg/ml and 1 µg/ml respectively while for clofazimine both were 1 µg/ml. The essential and categorical agreement for linezolid was 97.63% and 95.26% and for clofazimine, both were 100%. The sequencing result of the rplC gene revealed a point mutation at position 460 bp, where thymine (T) was substituted for cytosine (C) while seven mutations were noted between 46 to 220 bp in Rv0678 gene.ConclusionREMA method has been found to be more suitable in comparison to the agar dilution method due to lesser turnaround time. Mutations in rplC and Rv0678 genes were reasons for drug resistance against linezolid and clofazimine respectively.

Multicenter Italian Study on "In Vitro Activities" of Isavuconazole, Voriconazole, Amphotericin B, and Caspofungin for Aspergillus Species: Comparison between SensititreTM YeastOneTM and MIC Test Strip.

In this study the activity of Isavuconazole, Voriconazole, Amphotericin B, and Caspofungin against 224 clinical isolates of Aspergillus spp. originating from seven Italian hospitals, was comparatively evaluated with two commercial antifungal susceptibility tests (AST): SensititreTM YeastOneTM (SYO) and MIC Test Strip. More attention was focused on Isavuconazole activity, given the new introduction of the drug in widely distributed antifungal susceptibilities methods in the clinical microbiology lab. The minimum inhibitory concentrations of antifungal drug that can inhibit the growth of pathogen by 90% (MIC90) for Isavuconazole detected by SYO were 0.5, 1, 0.25, and 2 µg/mL for Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, and Aspergillus niger, respectively, whilst they were 0.25, 0.25, 0.5, and 0.75 µg/mL by MIC Test Strip. Essential agreement between the two tested methods for Isavuconazole is 70% for all the species tested, 75.7% for A. fumigatus, 45.2% for A. flavus, 90.6% for A. terreus, and 40% for A. niger. Although the tested strains do not express any phenotypic resistance, MIC results were quite different if tested with microdilution broth or gradient agar method. This is the first Italian multicenter report on Isavuconazole MIC obtained employing the widely used SensititreTM Yeast OneTM (SYO) and MIC Test Strip on clinical isolates of Aspergillus.

S2.5a Magnusiomyces spp.

S2.5 Rare yeasts, September 21, 2022, 3:00 PM - 4:30 PM Magnusiomyces clavatus and Magnusiomyces capitatus are emerging yeasts with intrinsic resistance to many commonly used antifungal agents. Identification is difficult, and the determination of susceptibility patterns with commercial and reference methods is equally challenging. For this reason, few data on invasive infections by Magnusiomyces spp. are available. We, therefore, determined the epidemiology and susceptibility of Magnusiomyces isolates from bloodstream infections (BSI) isolated in Germany and Austria from 2001 to 2020. A total of 34 Magnusiomyces BSI were analyzed; isolates were identified by internal transcribed spacer (ITS) sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Antifungal susceptibility was determined by EUCAST broth microdilution and gradient tests. Of the 34 isolates, M. clavatus was more common (n = 24) than M. capitatus (n = 10). BSI by Magnusiomyces spp. were more common in men (62%) and mostly occurred in patients with hemato-oncological malignancies (79%). The highest in vitro antifungal activity against M. clavatus/M. capitatus was observed for voriconazole (MIC50, 0.03/0.125 mg/l), followed by posaconazole (MIC50, 0.125/0.25 mg/l). M. clavatus isolates showed overall lower MICs than M. capitatus. With the exception of amphotericin B, a low essential agreement between gradient test and microdilution was recorded for all antifungals (0%-70%). Both species showed distinct morphologic traits on ChromAgar Orientation medium and Columbia blood agar, which can be used for differentiation if no MALDI-TOF MS or molecular identification is available. In conclusion, most BSI were caused by M. clavatus. The lowest MICs were recorded for voriconazole. Gradient tests demonstrated unacceptably low agreement and should preferably not be used for susceptibility testing of Magnusiomyces spp.

Impact of Alternative Growth Supplements on Antimicrobial Susceptibility Testing of Neisseria gonorrhoeae.

PurposeThe accurate detection of antibiotic susceptibility of Neisseria gonorrhoeae (N. gonorrhoeae) is of great importance for the treatment of patients with gonorrhea as well as to hinder the progress of drug resistance. To promote the application of gonococcal antibiotic susceptibility monitoring in primary hospitals and remote medical institutions, this study evaluated the effect of alternative growth supplements on the antibiotic susceptibility testing of N. gonorrhoeae isolates.MethodsWe divided the antimicrobial-containing media into three groups by adding different growth supplements (sterile defibrinated sheep blood, bovine hemoglobin, and Vitox). We tested the antimicrobial susceptibility of 80 N. gonorrhoeae isolates in different groups against eight antibiotics. Nonparametric signed-rank tests were utilized to compare the minimum inhibitory concentration (MIC) results of each group. Taking the MIC results of Vitox group as expected, the essential agreement (EA) and category agreement (CA) of the other two groups were calculated.ResultsFor the group using sheep blood as growth supplements, the EA values and CA values of each antibiotic were above 90.00% and minor error rates were less than 7.00%. No very major error and major error were observed. For the group using hemoglobin as growth supplements, the EA values of the susceptibility results of zoliflodacin, penicillin, and ceftriaxone were lower than 90.00%. The overall MIC results of using hemoglobin as a growth supplement were higher than those of sheep blood and Vitox in the susceptibility testing of these three antibiotics.ConclusionCompared with the expected results, sheep blood may be considered for the use as an alternative material for N. gonorrhoeae antibiotics susceptibility surveillance, while hemoglobin may not be suitable for supplement to antimicrobial-containing medium.

Evaluation Report of the Colistin Broth Disk Elution Method with Acinetobacter baumannii Isolates from a Low-Resource Setting.

ABSTRACTThe rapid emergence of drug resistance in Acinetobacter baumannii has put forward the use of colistin as a last-resort treatment for infections with A. baumannii. Empirical colistin use without prior susceptibility testing has been one of the factors that has been promoting drug resistance in low-resource settings. In this regard, while the advocated broth microdilution (BMD) method for colistin susceptibility testing is often considered cumbersome, the preferable colistin broth disk elution (CBDE) method has not yet been approved for A. baumannii. To prevent the underreporting of colistin susceptibility, we tested the CBDE method for A. baumannii and compared the results with those of BMD. A total of 125 A. baumannii, including 100 susceptible and 25 resistant isolates were tested via the CBDE method and compared with the standard BMD method. The essential agreement, categorical agreement, sensitivity, and specificity for CBDE were 97.6% (n = 122), 98.4% (n = 123), 100%, and 98.40%, respectively. The percentage of major error found was 1.6% (n = 2), and no very major error was found. CBDE in A. baumannii could be considered in low-resource settings.IMPORTANCE The relatively cumbersome broth microdilution (BMD) method for routine colistin susceptibility testing has not been adopted, especially in low-resource settings, often leading to the underreporting of colistin susceptibility and the promotion of the empirical use of colistin. In this regard, the much-preferred colistin broth disk elution (CBDE) method has not yet been approved for A. baumannii. We evaluated colistin susceptibility via the CBDE method, compared the results with those of the BMD method in 125 A. baumannii isolates with various profiles, and inferred that the CBDE method using 50 μL inoculum could be helpful, at least in resource-limited setups, versus not reporting susceptibility testing for colistin.

Quantitative analysis of Homoctenowakia bohemica bohemica (Tentaculitoids) from the Lochkovian of the Valle syncline, Ossa-Morena zone (SW Spain).

A quantitative study has been performed on a population of Homoctenowakia bohemica bohemica from the Ossa-Morena Zone (SW Spain). A comparison with a population from the Barrandian area shows essential agreement for all important characters, which fully conirms the specific assignement of the Spanish specimens.

Analysis of the FAST™ system for expedited identification and antimicrobial susceptibility testing of bloodborne pathogens

The high morbidity and mortality of sepsis can be impacted by expediting identification (ID) and antibiotic susceptibility testing (AST) of causative bacteria. We evaluated the Qvella FAST™ System which creates a Liquid Colony™ (LC) from blood cultures that can be used to expedite results by 24 to 48 hours. We analyzed 289 LC samples and found that there were 17 (5.9%) that resulted in no ID. One hundred percent of the LC samples that produced an ID were concordant with SOC identification. Gram-positive bacteria showed a categorical agreement (CA) of 99.5%, with 3 minor errors (minE), and no major errors (majE) or very major errors (VME), and essential agreement (EA) of 98.9%. For Gram-negatives, the CA was 97.8% and the EA was 98.5% with 31 minE, 0 majE, and 2 VME. The FAST-System™ can accelerate ID and AST by 24 to 48 hours with potential positive impacts on time to effective therapy for sepsis.

Susceptibility of Meropenem-Resistant and/or Carbapenemase-Producing Clinical Isolates of Enterobacterales (Enterobacteriaceae) and Pseudomonas aeruginosa to Ceftazidime-Avibactam and Ceftolozane-Tazobactam as Assessed by In Vitro Testing Methods.

This study aimed to assess the comparability of in vitro susceptibility testing methods to ceftazidime-avibactam (CZA) and ceftolozane-tazobactam (C/T). Meropenem-resistant and/or carbapenemase-producing clinical isolates of Enterobacterales (Enterobacteriaceae) and Pseudomonas aeruginosa were tested by both bioMérieux ETEST and VITEK-2 AST-N397 card and compared with a Micronaut AST-system broth microdilution (BMD) method. CZA and C/T MICs were interpreted using EUCAST breakpoints. Of the 153 Enterobacteriaceae isolates, 55.6% and 0.0% (VITEK 2) and 56.9% and 0.0% (ETEST and BMD) were susceptible to CZA and C/T, respectively. Of 52 P. aeruginosa isolates, 50.0% and 40.4% (VITEK 2, ETEST, and BMD) were susceptible to CZA and C/T, respectively. The essential agreement (EA) was 96.1% (197/205; VITEK 2 versus BMD) and 95.6% (196/205; ETEST versus BMD) for CZA testing, whereas EA was 98.0% (201/205; VITEK 2 versus BMD) and 96.6% (198/205; ETEST versus BMD) for C/T testing. The categorical agreement (CA) was 98.0% (201/205; VITEK 2 versus BMD) and 100% (ETEST versus BMD) for CZA testing, whereas CA was 100% (VITEK 2 versus BMD) and 100% (ETEST versus BMD) for C/T testing. Categorical errors regarded four Enterobacteriaceae isolates. VITEK 2 and ETEST yielded equivalent CZA and C/T susceptibility testing results, compared to the BMD method, in such a clinical context.

Comparative study of in vitro activity of tedizolid and linezolid against Mycobacterium avium complex.

ObjectivesThe aim of this study is to compare the in vitro activity and minimal inhibitory concentration (MIC) distributions of tedizolid and linezolid against Mycobacterium avium complex (MAC) strains using a reference broth microdilution assay and a macrodilution assay with the Bactec-MGIT-960. MethodsA total of 37 clinical isolates of MAC were included in the study. Reference broth microdilution was performed according to CLSI guidelines in a range of concentrations from 64 to 0.064 mg/L. Macrodilution was performed with the Bactec-MGIT-960 system. The cut-off points defined by CLSI for linezolid (resistant: > 16 mg/L, intermediate: 16 mg/L, susceptible: <16 mg/L) were used to define clinical categories of this drug. Essential agreement for both linezolid and tedizolid and categorical agreement for linezolid were defined following FDA criteria. ResultsThe MIC50 (16mg/L) and MIC90 (32mg/L) values for linezolid were identical with both methods. However, the MIC50 and MIC90 of tedizolid by microdilution (4 mg/L and 8 mg/L, respectively) were one twofold dilution higher than by macrodilution (2 mg/L and 4 mg/L, respectively). Ninety-four percent and 2.7% of the strains had MICs of tedizolid ≤4 mg/L and ≤ 0.5 mg/L, respectively, by the reference method. The linezolid macrodilution assay showed a categorical agreement of 40.5%, a minor error rate of 56.7% and a major error rate of 2.7% with respect to the reference method. ConclusionsTedizolid showed higher in vitro activity than linezolid against the tested MAC isolates. Macrodilution using the BD Bactec-MGIT-960 system is a practical approach to determine the susceptibility of MAC strains to tedizolid.

Intermittency of turbulent velocity and scalar fields using three-dimensional local averaging

An efficient approach for extracting 3D local averages in spherical subdomains is proposed and applied to study the intermittency of small-scale velocity and scalar fields in direct numerical simulations of isotropic turbulence. We focus on the inertial-range scaling exponents of locally averaged energy dissipation rate, enstrophy and scalar dissipation rate corresponding to the mixing of a passive scalar $\theta$ in the presence of a uniform mean gradient. The Taylor-scale Reynolds number $R_\lambda$ goes up to $1300$, and the Schmidt number $Sc$ up to $512$ (albeit at smaller $R_\lambda$). The intermittency exponent of the energy dissipation rate is $\mu \approx 0.23$, whereas that of enstrophy is slightly larger; trends with $R_\lambda$ suggest that this will be the case even at extremely large $R_\lambda$. The intermittency exponent of the scalar dissipation rate is $\mu_\theta \approx 0.35$ for $Sc=1$. These findings are in essential agreement with previously reported results in the literature. We further show that $\mu_\theta$ decreases monotonically with increasing $Sc$, either as $1/\log Sc$ or a weak power law, suggesting that $\mu_\theta \to 0$ as $Sc \to \infty$, reaffirming recent results on the breakdown of scalar dissipation anomaly in this limit.