Matrix Extension of the CompactDry™ "Nissui" YMR for Enumeration of Yeasts and Molds in Dried Cannabis Flower: AOAC Performance Tested MethodSM 092002.

BACKGROUND: CompactDry™ Yeast/Mold Rapid (YMR) is a ready-to-use dry media sheet using a chromogenic medium with selective agents for the enumeration of yeasts and molds in a variety of food products after incubation at 25 ± 1°C for 3 days. The method is certified as AOAC Performance Tested MethodSM 092002. OBJECTIVE: The CompactDry YMR method was validated for a matrix extension to cannabis flower through the AOAC Emergency Response Validation process. METHODS: The performance of the CompactDry YMR was compared to Dichloran Rose Bengal Chloramphenicol (DRBC) agar for the enumeration of yeasts and molds in cannabis flower. Matrix data were normalized by log10 transformation, and performance indicators included repeatability, difference of means (DOM), and inclusivity/exclusivity. RESULTS: The results demonstrated that the CompactDry YMR method is equivalent to the DRBC agar method at 72 h of incubation. In the independent laboratory validation study, there was no significant difference in detection, enumeration, or repeatability between the CompactDry YMR method and DRBC agar at 72 h. Eighteen inclusivity and 16 exclusivity strains specific to cannabis plant materials that were not evaluated in the original CompactDry YMR method validation were tested in this study. All inclusivity organisms produced typical colonies on the CompactDry YMR. The two exclusivity bacterial strains that showed growth on CompactDry YMR at 72 h were inoculated at a high concentration. CONCLUSIONS: CompactDry YMR is equivalent in performance to traditional culture media detection methods of yeasts and molds. HIGHLIGHTS: CompactDry YMR will streamline dried cannabis flower testing.

Evaluation of NG-Test Carba 5 for Rapid Phenotypic Detection and Differentiation of Five Common Carbapenemase Families: Results of a Multicenter Clinical Evaluation.

NG-Test Carba 5 is a rapid in vitro multiplex immunoassay for the phenotypic detection and differentiation of five common carbapenemase families (KPC, OXA-48-like, VIM, IMP, and NDM) directly from bacterial colonies. The assay is simple to perform and has recently received U.S. Food and Drug Administration clearance. A method comparison study was performed at geographically diverse medical centers (n = 3) in the United States, where 309 Enterobacterales and Pseudomonas aeruginosa isolates were evaluated by NG-Test Carba 5 (NG Biotech, Guipry, France), the Xpert Carba-R assay (Cepheid, Inc., Sunnyvale, CA), the modified carbapenem inactivation method (mCIM), the EDTA-modified carbapenem inactivation method, and disk diffusion with carbapenems. Colonies from tryptic soy agar with 5% sheep blood (blood agar) and MacConkey agar were tested, and the results were compared to those obtained by a composite reference method. Additionally, a fourth medical center performed a medium comparison study by evaluating the performance characteristics of NG-Test Carba 5 from blood, MacConkey, and Mueller-Hinton agars with 110 isolates of Enterobacterales and P. aeruginosa These results were compared to the expected genotypic and mCIM results. For the multicenter method comparison study, the overall positive percent agreement (PPA) and the overall negative percent agreement (NPA) of NG-Test Carba 5 with the composite reference method were 100% for both blood and MacConkey agars. The medium comparison study at the fourth site showed that the PPA ranged from 98.9% to 100% and that the NPA ranged from 95.2% to 100% for blood, MacConkey, and Mueller-Hinton agars. NG-Test Carba 5 accurately detected and differentiated five common carbapenemase families from Enterobacterales and P. aeruginosa colonies on commonly used agar media. The results of this test will support a streamlined laboratory work flow and will expedite therapeutic and infection control decisions.

The Continued Value of Disk Diffusion for Assessing Antimicrobial Susceptibility in Clinical Laboratories: Report from the Clinical and Laboratory Standards Institute Methods Development and Standardization Working Group.

Expedited pathways to antimicrobial agent approval by the U.S. Food and Drug Administration (FDA) have led to increased delays between drug approval and the availability of FDA-cleared antimicrobial susceptibility testing (AST) devices. Antimicrobial disks for use with disk diffusion testing are among the first AST devices available to clinical laboratories. However, many laboratories are reluctant to implement disk diffusion testing for a variety of reasons, including dwindling proficiency with this method, interruptions of the laboratory workflow, uncertainty surrounding the quality and reliability of disk diffusion tests, and a perceived need to report MIC values to clinicians. This minireview provides a report from the Clinical and Laboratory Standards Institute Methods Development and Standardization Working Group on the current standards and clinical utility of disk diffusion testing.

Comparative activities of cefepime and piperacillin/tazobactam tested against a global collection of Escherichia coli and Klebsiella spp. with an ESBL phenotype.

Cefepime exhibits more stability to hydrolysis by extended-spectrum beta-lactamases (ESBLs) compared with other cephalosporins, and piperacillin/tazobactam may be active against these pathogens because of the enzyme inhibitory activity of tazobactam. Thus, we evaluated the in vitro activity of these 2 antimicrobials against a large collection of isolates with an ESBL phenotype. A total of 50,637 clinical isolates (34,367 Escherichia coli and 16,270 Klebsiella spp.) collected from more than 80 medical centers (1998-2004) were tested by reference broth microdilution methods, and isolates with an ESBL phenotype (MIC, > or = 2 microg/mL for aztreonam or ceftazidime or ceftriaxone) were submitted to a clavulanate inhibition test (confirmation of ESBL production). Among isolates from North America, 3.9% of E. coli and 8.6% of Klebsiella spp. showed an ESBL phenotype, whereas among isolates from the rest of the world (ROW) (Europe, Latin America, and Asia), 7.7% of E. coli and 28.3% of Klebsiella spp. exhibited this pattern. Confirmation rates varied from 21.6% of E. coli in North America to 52.8% of Klebsiella spp. in the row Among E. coli from North America, cefepime (90.3% susceptibility) was generally more active than piperacillin/tazobactam (82.7%), especially among ESBL-not-confirmed (97.0% versus 85.5%). Cefepime also showed reasonable activity against Klebsiella spp. from North America (89.4% susceptibility). In general, isolates from North America exhibited higher susceptibility rates to both beta-lactams compared with isolates from the ROW, and ESBL-not-confirmed strains showed generally higher susceptibility rates than ESBL-confirmed organisms.

Antimicrobial susceptibility of Gram-negative bacteria in Brazilian hospitals: the MYSTIC Program Brazil 2003.

Establish the susceptibility pattern of Gram-negative bacteria causing infections in ICU patients, MYSTIC Program Brazil 2003. Gram-negative bacteria (n = 1,550) causing nosocomial infections were collected at 20 Brazilian centers. The central laboratory confirmed the identification and performed the susceptibility tests by Etest methodology (AB Biodisk, Solna, Sweden) for meropenem, imipenem, ciprofloxacin, ceftazidime, cefepime, cefotaxime, piperacillin/tazobactam, gentamicin, and tobramycin. Interpretation criteria used were according to National Committee for Clinical Laboratory Standards (NCCLS). Pseudomonas aeruginosa (30.3%) was the most frequent isolate, followed by E. coli (18.6%), Klebsiella pneumoniae (16.9%), Acitenobacter baumannii (8.8%), and Enterobacter cloacae (7.1%). Pseudomonas aeruginosa (n=470) isolates presented susceptibility rates of 64% to meropenem, 63.8% to piperacillin/tazobactam, 63.4% to amikacin, 58.7% to imipenem. Acitenobacter baumannii presented susceptibility rates to meropenem of 97.1%, and 73% to tobramycin. E. coli and K. pneumoniae were highly susceptible to both carbapenems. Carbapenem resistance among the Enterobacteriaceae is still rare in the region. Acitenobacter baumannii and P. aeruginosa presented elevated resistance rates to all antimicrobials. Since they play an important role in nosocomial infections in this environment, the use of empirical combination therapy to treat these pathogens may be justified.

In vitro activity of tigecycline against 3989 Gram-negative and Gram-positive clinical isolates from the United States Tigecycline Evaluation and Surveillance Trial (TEST Program; 2004).

The Tigecycline Evaluation and Surveillance Trial (TEST Program) determined the in vitro activity of tigecycline over a large population of organisms from geographically diverse sites. Tigecycline was compared to amikacin, ampicillin, amoxicillin/clavulanic acid, imipenem, cefepime, ceftazidime, ceftriaxone, levofloxacin, minocycline, piperacillin/tazobactam, linezolid, penicillin, and vancomycin against 3989 commonly encountered clinical Gram-negative and Gram-positive pathogens collected from sites in the United States during 2004. The tigecycline activity was equivalent to imipenem against Enterobacteriaceae. Tigecycline inhibited extended-spectrum beta-lactamase and AmpC phenotypes at MIC90 values (minimum inhibitory concentration) of < or =2 microg/mL. In vitro results for tigecycline were similar to other broad-spectrum antimicrobial agents against nonfermenters with MIC90 results of 2 microg/mL against Acinetobacter spp. and >16 microg/mL against Pseudomonas aeruginosa. Tigecycline demonstrated potent activity against Staphylococcus aureus (MIC90, 0.25 microg/mL) and enterococci (MIC90, 0.12 microg/mL) regardless of methicillin or vancomycin susceptibility. Tigecycline MIC values were unaffected by penicillin nonsusceptibility and beta-lactamase production among fastidious respiratory pathogens (Streptococcus pneumoniae [MIC90, 0.5 microg/mL] and Haemophilus influenzae [MIC90, 0.25 microg/mL]). Tigecycline offers excellent activity against most of the commonly encountered nosocomial and community-acquired bacterial pathogens.

Antimicrobial susceptibility of Gram-negative bacteria in Brazilian hospitals: the MYSTIC Program Brazil 2003

Establish the susceptibility pattern of Gram-negative bacteria causing infections in ICU patients, MYSTIC Program Brazil 2003. Gram-negative bacteria (n = 1,550) causing nosocomial infections were collected at 20 Brazilian centers. The central laboratory confirmed the identification and performed the susceptibility tests by Etest methodology (AB Biodisk, Solna, Sweden) for meropenem, imipenem, ciprofloxacin, ceftazidime, cefepime, cefotaxime, piperacillin/tazobactam, gentamicin, and tobramycin. Interpretation criteria used were according to National Committee for Clinical Laboratory Standards (NCCLS). Pseudomonas aeruginosa (30.3 percent) was the most frequent isolate, followed by E. coli (18.6 percent), Klebsiella pneumoniae (16.9 percent), Acitenobacter baumannii (8.8 percent), and Enterobacter cloacae (7.1 percent). Pseudomonas aeruginosa (n=470) isolates presented susceptibility rates of 64 percent to meropenem, 63.8 percent to piperacillin/tazobactam, 63.4 percent to amikacin, 58.7 percent to imipenem. Acitenobacter baumannii presented susceptibility rates to meropenem of 97.1 percent, and 73 percent to tobramycin. E. coli and K. pneumoniae were highly susceptible to both carbapenems.Carbapenem resistance among the Enterobacteriaceae is still rare in the region. Acitenobacter baumannii and P. aeruginosa presented elevated resistance rates to all antimicrobials. Since they play an important role in nosocomial infections in this environment, the use of empirical combination therapy to treat these pathogens may be justified.

Infecções do trato respiratório: principais agentes bacterianos e padrões de resistência Dados brasileiros do estudo internacional PROTEKT / Respiratory tract infections: major bacterial pathogens and patterns of resistance. Brazilian results from PROTEKT surveillance study

As infecções da trato respiratório (ITR) são responsáveis por significativa morbidade e mortalidade. Devido a sua diversificada etiologia e dificuldades diagnósticas, muitas vezes, o tratamento destas infecções é iniciado empiricamente.

Atividade antimicrobiana in vitro de quinupristina/dalfopristina para cocos gram-positivos isolados de cinco centros brasileiros: resultado do estudo de vigilância L-SMART / Antimicrobial in vitro activity of quinupristin/dalfopristin against gram-positive cocci isolated from 5 Brazilian centers: results from the local smart (L-SMART) surveillance study

Há alguns anos tem-se verificado um aumento progressivo da resistência de alguns cocos gram-positivos a determinados antimicrobianos. Este aumento da resistência tem sido observado principalmente no ambiente hospitalar, e as bactérias mais comumente envolvidas são os Staphylococcus spp. e os Enterococcus spp. Devido a este fato, novos antimicrobianos são avaliados para o tratamento de infecções causadas por estas cepas multirresistentes. A associação quinupristina/dalfopristina (Q/D), também conhecida como Synercid©, é um antibacteriano da classe das estreptograminas, de uso endovenoso, composto por dois derivados semi-sintéticos da pristanamicina. A combinação das estreptograminas B e A na razão de 30:70 tem atividade antimicrobiana voltada para cocos gram-positivos, como Staphylococcus spp., Streptococcus spp., incluindo S. pneumoniae e Enterococcus faecium, sendo o E. faecalis habitualmente resistente. Neste estudo foi avaliada atividade in vitro de Q/D e outros oito antimicrobianos frente a 631 amostras de cocos gram-positivos isoladas de cinco centros brasileiros, complementadas com outras 20 cepas de E. faecium resistentes à vancomicina, provenientes dos Estados Unidos. Para a avaliação da sensibilidade aos antimicrobianos foi determinada a concentração inibitória mínima (MIC) pelo método do Etest (AB Biodisk, Solna, Suécia) e as cepas testadas foram: Staphylococcus aureus (n=267), Staphylococcus coagulase negativo (n=131), Streptoccus pneumoniae (n=130), Streptococcus beta-hemolíticos (n=28), Enteroccus faecalis (n=44) e E. faecium (n=51). A Q/D demonstrou excelente atividade contra Staphylococcus spp., independente de serem sensíveis ou resistentes à oxacilina. Para S. pneumoniae, a Q/D apresentou igualmente uma ótima atividade, inclusive para as cepas com resistência intermediária ou total para penicilina. Entre as cepas de E. faecium sensíveis à vancomicina, o MIC 90 de Q/D obtido foi de 3µg/ml, sendo que 45 por cento das cepas testadas foram sensíveis e 55 por cento apresentaram sensibilidade intermediária à associação. Desta forma, pode-se afirmar que a associação. Desta forma, pode-se afirmar que a associação Q/D representa uma nova opção para o tratamento endovenoso de infecções causadas por cocos gram-positivos, principalmente para as cepas multiresistentes, sendo também uma alternativa ao uso de glicopeptídeos

Avaliaçäo in vitro de macrolídeos e antibióticos B-lactâmicos orais ante a microrganismos isolados no tato respiratório / In vitro evaluation of oral macrolides and B-lactam antibiotics against isolated Pathogens from the respiratory tract

No período de setembro de 1999 a julho de 2000, analisamos a atividade in vitro de seis antimicrobianos, respectivamente, azitromicina, claritromicina, amoxicilina, amoxicilina/ácido clavulânico, cefprozil e cefaclor ante a um total de 597 amostras bacterianas isoladas do trato respiratório superior e inferior de pacientes da comunidade, sem restriçäo de faixa etária, assim distribuídas: 247 cepas de H. influenzae, 147 S. aureus, 114 S. pneumoniae, 51 S. pyogenes e 38 M catarrhalis. A determinaçäo da Concentraçäo Inibitória Mínima (MIC) foi realizada pelo método do Etest e interpretadas de acordo com critérios padronizados pelo NCCLS. Entre as 247 cepas de H. influenzae, 9,7 porcento eram produtoras de B-lactamase, sendo que 100 porcento destas cepas foram sensíveis à amoxicilina/ác. clavulânico e cefaclor, 98 porcento à cefprozil, 96 porcento à azitromicina e 90 porcento à claritromicina. Das 147 cepas de S. aureus, 100 porcento eram sensíveis à oxacilina, amoxilina/ác. clavulânico, cefprozil e cefaclor, 71 porcento à claritromicina e 68 porcento à azitromicina. Entre as 114 cepas de S. peneumoniae, nenhuma cepa apresentou resistência total à penicilina, 14 cepas apresentaram resistência intermediária para penicilina, sendo que 100 porcento das cepas foram sensíveis à amoxicilina e amoxicilina/àc. clavulânico, 99 porcento à cefprozil e cefaclor, 88 porcento à azitromicina e 86 porcento à claritromicina. Entre as 14 cepas de pneumococos com resistência intermediária para penicilina, 3 cepas eram resistentes à azitromicina e 4 cepas à claritromicina. Entrre as 51 amostras de S. pyogenes, 4 cepas apresentaram resistência à azitromicina e claritromicina. Para M. catarrhalis, 100 porcento das cepas eram produtoras de B-lactamase e o MIC para azitromicina, claritromicina, amoxicilina, amoxicilina/ác. clavulânico, cefprozil e cefaclor foram 0,25; 0,25; 3; 0,25; 4 e 1,5 ug/ml, respectivamente.(au)

Detecçäo e prevalência de ß-lactamases de espectro ampliado (ESBL) em amostras bacterianas isoladas em hospitais brasileiros / Detection and prevalence of extended-spectrum ß-lactamases (ESBL) in bacteria isolated from Brazilian hospitals

A resitência aos antibióticos ß-lactâmicos, principalmente em membros da família Enterobacteriaceae, tem aumentado significativamente nos últimos anos, particularmente em algumas espécies, como Klebsiella pneumoniae e Escherichia coli. No ambiente hospitalar, destacam-se as infecçöes causadas por enterobactérias produtoras de ß-lactamases de espectro ampliado (ESBL). As bactérias produtoras destas enzimas apresentam um grande desafio tanto para sua detecçäo laboratorial, quanto para o tratamento adequado das infecçöes por elas causadas. Neste estudo, avalianos a produçäo de ESBL em 480 cepas bacterianas (269 E. coli e 211 K. pneumoniae), isoladas em 12 hospitais e laboratórios brasileiros, utilizando normas preconizadas pelo NCCLS, complementadas pela metodologia Etest (AB Biodisk). Obtivemos uma percentagem alarmante de cepas produtoras de ESBL, principalmente para K. pneumoniae, onde 44,1 por cento (93/211) das mesmas eram positivas, enquanto que somente 5,6 por cento (15/269) das amostras de E. coli eram produtoras de ESBL. O objetivo deste estudo foi documentar a prevalência, em hospitais brasileiros, de amostras de K. pneumoniae e E. coli produtoras de ESBL, enfatizando a importância da correta realizaçäo, leitura e interpretaçäo do antibiograma, além da realizaçäo de testes confirmatórios para sua detecçäo

Evaluation of the in vitro activity of 9 antimicrobials agains bacterial strains isolated from patients in intensive care units in Brazil: MYSTIC antimicrobial surveillance program

Multi-resistant bacterial strains are increasingly prevalent in hospital environments. Bacterial resistance is an important problem, especially for practitioners in intensive care units (ICUs) because of the selective pressure on the prevalente bacteria in these environments. The MYSTIC (Meropenen Yearly Susceptibility Test Information Collection) study has been monitoring the performance of carbapenems and otherantibiotics in different hospitals for at least 3 years. The in vitro activities of meropenem, imipenem, ceftazidime, cefepime, cefotaxime, ciprofloxacin, piperacilin/tazobactam, gentamicin, and tobramycin were compared against 452 recent clinical aerobic isolates. The isolates consisted of 19 species of Gram-negative bacteria (n=290) including K. pneumoniae (n=49), E. coli (n=48), A. baumannii (n=47), Enterobacter spp. (n=41), and P. aeruginosa (n=33) and 9 species of Gram-positive bacteria (n=162) including Staphylococcus aureus (n=63), Enterococcus faecalis (n=22), Streptococcus pneumoniae (n=22) and coagulase negative Staphylococci (n=21). All isolates were collected from ICU patients. Minimal inhhibitory concentrations (MICs) were determined by Etest methodology, using standardized and controlled procedures. Meropenem and imipenem showed the lowest MIC for all species tested. Gram-negative isolates showed the following overall resistance percentages to the other 7 drugs: tobramycin (43.1 por cento), cefotaxime (38.6 por cento), gentamicin (34.1 por cento), ceftazidime (31.7 por cento), ciprofloxacin (25.5 por cento), piperacillin/tazobactam (26.9 por cento), and cefepime (18.6 por cento). Carbapenems were the most active drugs overall and only P. aeruginosa presented some degree of resistance(18.2 por cento). We also evaluated the production of extended spectrum B-lactamase (ESBL) among all Enterobacteriaceae members (n=176) by Etest/ESBL, strip. ESBL production was detected in 51 strains (29.0 por cento). Among them, Klebsiella pneumoniae was the most prevalent at 59.2 porcento, followed by Enterobacter spp (19.5 porcento) and E. coli (14.6 por cento). The high level of resistance against several antimicrobialsand the alarming rate of ESBL production may restrict therapeutic choice to the carbapenems in this selected group of patients.