Comparison of disk diffusion,Etest and VITEK2 for detection of carbapenemase‐producing Klebsiella pneumoniae with the EUCAST and CLSI breakpoint systems

The aim of this study was to compare CLSI and EUCAST MIC and disk diffusion carbapenem breakpoints for the detection of carbapenemase‐producing Klebsiella pneumoniae. K. pneumoniae strains with known KPC (n = 31) or VIM (n = 20) carbapenemases were characterized by disk diffusion (Oxoid) and Etest (bioMérieux) vs. imipenem, meropenem and ertapenem, and with VITEK2 (bioMérieux, five different cards). Extended‐spectrum β‐lactamase (ESBL) testing was performed with VITEK2 (bioMérieux), ESBL combination disks (Becton Dickinson) and the ESBL Etest (bioMérieux). With CLSI and EUCAST MIC breakpoints, respectively, 11 and seven of the strains were susceptible to imipenem, 12 and eight to meropenem, and seven and none to ertapenem. The EUCAST epidemiological cut‐off (ECOFF) values for meropenem and ertapenem identified all carbapenemase producers, whereas the imipenem ECOFF failed in five strains. All carbapenemase producers were detected with EUCAST disk diffusion breakpoints for ertapenem and meropenem, and four strains were susceptible to imipenem. CLSI disk diffusion breakpoints characterized 18 (imipenem), 14 (meropenem) and three (ertapenem) isolates as susceptible. When cards with a single carbapenem were used, detection failures with VITEK2 were four for imipenem, none for meropenem and one for ertapenem. Cards containing all three carbapenems had one to two failures. With ESBL combination disks, 21/31 KPC producers and 2/20 VIM producers were positive. With VITEK2, no VIM producers and between none and seven KPC producers were ESBL‐positive. All carbapenemase producers were detected with the meropenem MIC ECOFF, or the clinical EUCAST breakpoint for ertapenem. EUCAST disk diffusion breakpoints for meropenem and ertapenem detected all carbapenemase producers. VITEK2 had between none and four failures in detecting carbapenemase producers, depending on the antibiotic card.     

Multidrug-Resistant Candida auris Misidentified as Candida haemulonii: Characterization by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry and DNA Sequencing and Its Antifungal Susceptibility Profile Variability by Vitek 2, CLSI Broth Microdilution,and Etest Method

Candida auris is a multidrug-resistant yeast that causes a wide spectrum of infections, especially in intensive care settings. We investigated C. auris prevalence among 102 clinical isolates previously identified as Candida haemulonii or Candida famata by the Vitek 2 system. Internal transcribed spacer region (ITS) sequencing confirmed 88.2% of the isolates as C. auris, and matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) easily separated all related species, viz., C. auris (n = 90), C. haemulonii (n = 6), C. haemulonii var. vulnera (n = 1), and Candida duobushaemulonii (n = 5). The in vitro antifungal susceptibility was determined using CLSI broth microdilution (CLSI-BMD), the Vitek 2 antifungal susceptibility test, and the Etest method. C. auris isolates revealed uniformly elevated fluconazole MICs (MIC₅₀, 64 μg/ml), and an alarming percentage of isolates (37%) exhibited elevated caspofungin MICs by CLSI-BMD. Notably, 34% of C. auris isolates had coexisting elevated MICs (≥2 μg/ml) for both fluconazole and voriconazole, and 10% of the isolates had elevated coexisting MICs (≥2 μg/ml) to two additional azoles, i.e., posaconazole and isavuconazole. In contrast to reduced amphotericin B MICs by CLSI-BMD (MIC₅₀, 1 μg/ml) for C. auris, elevated MICs were noted by Vitek 2 (MIC₅₀, 8 μg/ml), which were statistically significant. Candida auris remains an unnoticed pathogen in routine microbiology laboratories, as 90% of the isolates characterized by commercial identification systems are misidentified as C. haemulonii. MALDI-TOF MS proved to be a more robust diagnostic technique for rapid identification of C. auris. Considering that misleading elevated MICs of amphotericin B by the Vitek AST-YS07 card may lead to the selection of inappropriate therapy, a cautionary approach is recommended for laboratories relying on commercial systems for identification and antifungal susceptibility testing of rare yeasts.     

Value of (1-3)-β-d-glucan, Candida mannan and Candida DNA detection in the diagnosis of candidaemia

This study determined the value of (1,3)-β-d-glucan (BDG), Candida mannan (MN) and Candida species-specific DNA as surrogates for diagnosis of candidaemia. Thirty-nine patients yielding Candida species in blood cultures were investigated for presence of BDG, MN and Candida species-specific DNA in serum samples. The Candida spp. bloodstream isolates included C. albicans (n = 16), C. tropicalis (n = 10), C. parapsilosis (n = 7), C. glabrata (n = 3) and C. dubliniensis (n = 3). Positivity of the three markers was as follows: Candida DNA for corresponding Candida species, 100%; BDG, 87%; MN, 59%. Despite varying sensitivities of these biomarkers, they provided a useful adjunct to the diagnosis of candidaemia.     

Assessment of the Phoenix™ automated system and EUCAST breakpoints for antimicrobial susceptibility testing against isolates expressing clinically relevant resistance mechanisms

EUCAST breakpoint criteria are being adopted by automatic antimicrobial susceptibility testing systems. The accuracy of the Phoenix Automated System in combination with 2012 EUCAST breakpoints against recent clinical isolates was evaluated. A total of 697 isolates (349 Enterobacteriaceae, 113 Pseudomonas spp., 25 Acinetobacter baumannii, 11 Stenotrophomonas maltophilia, 95 Staphylococcus aureus, 6 coagulase negative staphylococci, 77 enterococci and 21 Streptococcus pneumoniae) with defined resistance phenotypes and well-characterized resistance mechanisms recovered in Spain (n = 343) and Italy (n = 354) were tested. Comparator antimicrobial susceptibility testing data were obtained following CLSI guidelines. Experimental agreement (EA), defined as MIC agreement ±1 log₂ dilution, category agreement (CA) and relative discrepancies (minor (mD), major (MD) and very major discrepancies (VMD)) were determined. The overall EA and CA for all organism-antimicrobial agent combinations (n = 6.294) were 97.3% and 95.2%, respectively. mD, MD and VMD were 4.7%, 1.3% and 2.7%, all of them in agreement with the ISO (ISO20776–2:2007) acceptance criteria for assessment of susceptibility testing devices. VMD were mainly observed in amoxicillin-clavulanate and cefuroxime in Enterobacteriaceae and gentamicin in Pseudomonas aeruginosa, whereas MD were mainly observed in amoxicillin-clavulante in Enterobacteriaceae. mD were mainly observed in Enterobacteriaceae but distributed in different antimicrobials. For S. aureus and enterococci relative discrepancies were low. The Phoenix system showed accuracy assessment in accordance with the ISO standards when using EUCAST breakpoints. Inclusion of EUCAST criteria in automatic antimicrobial susceptibility testing systems will facilitate the implementation of EUCAST breakpoints in clinical microbiology laboratories.     

ΔF659 and F659S substitutions at the HS1 of FKS2 gene,along with E655A and W715L upstream and downstream substitutions,correlate with high ibrexafungerp MICs against Candida glabrata

ObjectivesIbrexafungerp is a new inhibitor of Candida spp glucan synthase. We previously set the ibrexafungerp wild-type upper limit (wtUL) against Candida glabrata. We here assessed which FKS2 gene substitutions confer an ibrexafungerp non-wild-type phenotype in C. glabrata isolates.MethodsWe studied a set of C. glabrata (n = 34) isolates showing resistance to micafungin and anidulafungin (n = 28) or only to anidulafungin (n = 6) and harbouring 10 different FKS2 gene substitutions. Antifungal susceptibility to ibrexafungerp was tested according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) E.Def 7.3.2 procedure and isolates were considered ibrexafungerp non-wild type according to the statistical wtUL (minimum inhibitory concentration [MIC] ≥2) or visual wtUL (MIC ≥4).ResultsIbrexafungerp MICs against the isolates ranged from 0.06 to 4 mg/L. Four FKS2 gene substitutions (ΔF659, F659S, E655A, and W715L) were exclusively found in isolates showing an ibrexafungerp MIC above the statistical wtUL (≥2 mg/L) whereas isolates harbouring other substitutions were found to be ibrexafungerp wild type. The use of the visual wtUL (MIC ≥4 mg/L) bisected the population of isolates harbouring such substitutions.DiscussionC. glabrata isolates showing an ibrexafungerp MIC ≥2 mg/L may be considered non-wild type and are prone to harbour ΔF659, F659S, E655A, and W715L substitutions at the FKS2 gene. It is worth noting that substitutions ΔF659 and F659S were located at the beginning of the HS1 of FKS2 gene of C. glabrata. The role of other substitutions on conferring a non-wild-type phenotype to ibrexafungerp is not well elucidated.     

Candida guilliermondii and Other Species of Candida Misidentified as Candida famata: Assessment by Vitek 2, DNA Sequencing Analysis,and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry in Two Global Antifungal Surveillance Programs

Candida famata (teleomorph Debaryomyces hansenii) has been described as a medically relevant yeast, and this species has been included in many commercial identification systems that are currently used in clinical laboratories. Among 53 strains collected during the SENTRY and ARTEMIS surveillance programs and previously identified as C. famata (includes all submitted strains with this identification) by a variety of commercial methods (Vitek, MicroScan, API, and AuxaColor), DNA sequencing methods demonstrated that 19 strains were C. guilliermondii, 14 were C. parapsilosis, 5 were C. lusitaniae, 4 were C. albicans, and 3 were C. tropicalis, and five isolates belonged to other Candida species (two C. fermentati and one each C. intermedia, C. pelliculosa, and Pichia fabianni). Additionally, three misidentified C. famata strains were correctly identified as Kodomaea ohmeri, Debaryomyces nepalensis, and Debaryomyces fabryi using intergenic transcribed spacer (ITS) and/or intergenic spacer (IGS) sequencing. The Vitek 2 system identified three isolates with high confidence to be C. famata and another 15 with low confidence between C. famata and C. guilliermondii or C. parapsilosis, displaying only 56.6% agreement with DNA sequencing results. Matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) results displayed 81.1% agreement with DNA sequencing. One strain each of C. metapsilosis, C. fermentati, and C. intermedia demonstrated a low score for identification (<2.0) in the MALDI Biotyper. K. ohmeri, D. nepalensis, and D. fabryi identified by DNA sequencing in this study were not in the current database for the MALDI Biotyper. These results suggest that the occurrence of C. famata in fungal infections is much lower than previously appreciated and that commercial systems do not produce accurate identifications except for the newly introduced MALDI-TOF instruments.     

Clinical manifestations of candidemia caused by uncommon Candida species and antifungal susceptibility of the isolates in a regional hospital in Taiwan, 2007–2014

ObjectiveThis retrospective study investigated clinical manifestations of candidemia caused by uncommon Candida species and antifungal susceptibility of the isolates in a regional hospital in Taiwan.MethodsThe uncommon Candida species was initially defined as Candida species other than C. albicans, C. tropicalis, C. glabrata complex, C. parapsilosis complex and C. krusei. All uncommon Candida isolates were identified and confirmed by molecular methods. In vitro susceptibility testing of the uncommon Candida species to nine antifungal agents was conducted using the broth microdilution method with the Sensititre YeastOne (SYO) system (Trek Diagnostic Systems, Ltd., East Grimstead, UK).ResultsTwenty-one patients, comprising 11 males and 10 females with a median age of 69 years, were recruited. Cancer (n = 11) was the most common underlying disease, 19 (90.5%) cases had prior antibiotic exposure, and only two patients had prior antifungal use. The overall in-hospital mortality rate was 38.1% (n = 8). C. guilliermondii (n = 11) was the most common pathogen, followed by C. curvata (n = 3). C. guilliermondii isolates exhibited relatively high rates of azole minimum inhibitory concentrations (MICs) above epidemiological cut-off values (ECVs), whereas C. pelliculosa and C. lusitaniae isolates all remained susceptible to azoles. All three C. curvata isolates had high caspofungin (>8 mg/L) and fluconazole MICs (8 mg/L) and could be defined as multidrug-resistant.ConclusionsUncommon Candida species frequently exhibit high rates of non-susceptibility to antifungals. Identification of all Candida isolates at the species level from blood samples is of value for treatment.     

Efficient capture of Candida albicans and zymosan by SIGNR1 augments TLR2-dependent TNF-α production

SIGNR1, a mouse C-type lectin, binds various pathogens, including Candida albicans. In this study, we explore the impact of SIGNR1 in the recognition of C. albicans/zymosan and the subsequent tumor necrosis factor (TNF)-α production using SIGNR1-transfected RAW264.7 (RAW-SIGNR1) cells and resident peritoneal macrophages. Compared with RAW-control cells, RAW-SIGNR1 cells dramatically enhanced TNF-α production upon the stimulation with heat-killed C. albicans and zymosan. Recognition of microbes via carbohydrate recognition domain (CRD) of SIGNR1 was crucial for the enhanced TNF-α production. Consistently, such an enhancement was significantly decreased by anti-SIGNR1 mAb. Laminarin, antagonistic Dectin-1 ligand, cooperated to further diminish the response, although no effect was observed by itself in RAW-SIGNR1 cells. However, it moderately reduced the response of RAW-control cells. Zymosan depleted of toll-like receptor (TLR) ligands decreased the response, even though it was recognized by SIGNR1 and Dectin-1. Moreover, antagonistic anti-TLR2 abolished the response, suggesting that TNF-α production largely relies on TLR2-mediated signaling. Resident peritoneal macrophages expressing SIGNR1 predominantly captured zymosan injected intra-peritoneally and produced TNF-α, which was dependent on TLR2 and partly inhibited by anti-SIGNR1 mAb. Finally, physical association of SIGNR1 with the extracellular portion of TLR2 through CRD was confirmed by immunoprecipitation using various deletion mutants. These results suggest that SIGNR1 recognizing microbes participates in the enhanced TNF-α production by M? in cooperation with TLR2.     

Potential effect of 2-isopropyl-5-methylphenol (thymol) alone and in combination with fluconazole against clinical isolates of Candida albicans,C. glabrata and C. krusei

Limitations of antifungals used in the treatment of candidiasis, as the development of resistant strains, are known by the scientific community. In this context, the aim of this study was to investigate the activity of 2-isopropyl-5-methylphenol (thymol) in combination with fluconazole (FLZ) against clinical Candida strains. The antifungal activity of thymol along with FLZ was evaluated by the Clinical Laboratory Standards Institute (CLSI) M27-A2 broth microdilution method. In addition, synergism was observed for clinical strains of Candida spp. with combination of thymol-FLZ evaluated by the chequerboard microdilution method. The mean of minimum inhibitory concentration (MIC) values of thymol and FLZ were 49.37 and 0.475 μg/ml for C. albicans, 51.25 and 18.80 μg/ml for C. glabrata and 70 and 179.20 μg/ml for C. krusei strains, respectively. Thymol in combination with FLZ exhibited the synergistic effects against all species of Candida tested. FICI values for thymol plus FLZ ranged from 0.366 to 0.607 for C. albicans strains, 0.367 to 0.482 for C. glabrata strains, and 0.375 to 0.563 for C. krusei strains. No antagonistic activity was seen in the strains tested. Thymol was found to have a fungicidal effect on Candida species and a synergistic effect when combined with FLZ.     

Comparison of disc and MIC reduction methods with polymerase chain reaction for the detection of metallo-β-lactamase in Pseudomonas aeruginosa

Purpose: The present study was undertaken to evaluate the screening antibiotic, confirmatory phenotypic test and agent against PCR as gold standard and to detect the prevalent MBL gene. Materials and Methods: Three hundred and twenty-six Pseudomonas aeruginosa isolates were screened for resistance to Imipenem (IPM), Meropemem (MEM) and Ceftazidime (CAZ) by disc diffusion. Isolates resistant to any of these were considered screen test-positive for MBL and were subjected to Double disc synergy test (DDST) and Disc potentiation test (DPT: Using IPM, MEM and CAZ alone and with EDTA), Minimum inhibitory concentration (MIC) reduction [four-fold or more reduction in MIC of IPM and MEM in presence of chelators: EDTA and 1,10-phenanthroline (EPI/EPM: EDTA-phenanthroline- Imipenem/Meropenem Broth Microdilution method)] and polymerase chain reaction (PCR) for bla_IMP and bla_VIM. Results: Screen test-positives by MEM and CAZ were 19.3% as against 17.8% by IPM. MEMDDST, DPT and EPM confirmed 100% screen-test positives as against 93.7% by CAZ DDST and DPT-2, 76.2% by CAZ DPT-1, 88.9% by IPM DDST, 85.7% by IPM DPT-1 and 92.1% by EPI. IPMand CAZ DDST together confirmed 100% while IPM and CAZ DPT-2 confirmed 96.8%. All 63 screen-test positives showed the presence of bla_VIM. Conclusions: MEM was found to be the best screening and confirmatory agent for MBL detection and bla_VIM was found to be the prevalent MBL gene in this part of the country.     

φ(2)GFP10, a high-intensity fluorophage, enables detection and rapid drug susceptibility testing of Mycobacterium tuberculosis directly from sputum samples

The difficulty of diagnosing active tuberculosis (TB) and lack of rapid drug susceptibility testing (DST) at the point of care remain critical obstacles to TB control. This report describes a high-intensity mycobacterium-specific-fluorophage (φ(2)GFP10) that for the first time allows direct visualization of Mycobacterium tuberculosis in clinical sputum samples. Engineered features distinguishing φ(2)GFP10 from previous reporter phages include an improved vector backbone with increased cloning capacity and superior expression of fluorescent reporter genes through use of an efficient phage promoter. φ(2)GFP10 produces a 100-fold increase in fluorescence per cell compared to existing reporter phages. DST for isoniazid and oxofloxacin, carried out in cultured samples, was complete within 36 h. Use of φ(2)GFP10 detected M. tuberculosis in clinical sputum samples collected from TB patients. DST for rifampin and kanamycin from sputum samples yielded results after 12 h of incubation with φ(2)GFP10. Fluorophage φ(2)GFP10 has potential for clinical development as a rapid, sensitive, and inexpensive point-of-care diagnostic tool for M. tuberculosis infection and for rapid DST.     

Is there a correlation between the structure of hair and breast cancer or BRCA1/2 mutations?

It has been suggested that the small angle x-ray scattering (SAXS) pattern of human hair can be used to diagnose breast cancer and possibly to identify BRCA1/2 mutation carriers, who are at significantly elevated risk for developing breast cancer. In particular, the presence of a diffuse ring in the SAXS pattern was said to be diagnostic of either breast cancer or an increased risk thereof. To test this hypothesis, we measured SAXS from the pubic hair of 56 subjects with known BRCA1/2 and breast cancer status. We found that there is no clear association between the pattern of SAXS seen in human pubic hair and the risk of breast cancer or the presence of BRCA1/2 mutations. The possible use of SAXS to diagnose cancer remains conjectural, but this and previous studies do not suggest that SAXS can be used as a reliable method of identifying either BRCA1/2 mutation carriers or women who have had breast cancer.     

Aims,design and methods of a case–control study for the assessment of the role of dietary habits,eating behaviors and environmental factors,on the development of breast cancer

In this work the aims, methodology and procedures of a case–control study that was developed for assessing the role of dietary habits, eating behaviors and environmental factors on the development of breast cancer, is presented. During 2010–2012, 250 consecutive women patients, newly diagnosed with breast cancer and 250 population-based, healthy subjects (controls) age-matched to the cases, were enrolled. Socio-demographic, dietary, psychological, lifestyle as well as environmental characteristics (i.e., exposure to pollution, pesticides, electromagnetic fields and radon) were recorded through face-to-face interviews with the participants. Dietary habits and eating behaviors were evaluated with a special questionnaire that had been developed for the study, and was found relatively valid and reliable.     

Spectrum of MEK1 and MEK2 gene mutations in cardio-facio-cutaneous syndrome and genotype�Cphenotype correlations

Cardio-facio-cutaneous syndrome (CFCS) is a rare disease characterized by mental retardation, facial dysmorphisms, ectodermal abnormalities, heart defects and developmental delay. CFCS is genetically heterogeneous and mutations in the KRAS, BRAF, MAP2K1 (MEK1) and MAP2K2 (MEK2) genes, encoding for components of the RAS–mitogen activated protein kinase (MAPK) signaling pathway, have been identified in up to 90% of cases. Here we screened a cohort of 33 individuals with CFCS for MEK1 and MEK2 gene mutations to further explore their molecular spectrum in this disorder, and to analyze genotype–phenotype correlations. Three MEK1 and two MEK2 mutations were detected in six patients. Two missense MEK1 (L42F and Y130H) changes and one in-frame MEK2 (K63_E66del) deletion had not been reported earlier. All mutations were localized within exon 2 or 3. Together with the available records, the present data document that MEK1 mutations are relatively more frequent than those in MEK2, with exons 2 and 3 being mutational hot spots in both genes. Mutational analysis of the affected MEK1 and MEK2 exons did not reveal occurrence of mutations among 75 patients with Noonan syndrome, confirming the low prevalence of MEK gene defects in this disorder. Clinical review of known individuals with MEK1/MEK2 mutations suggests that these patients show dysmorphic features, ectodermal abnormalities and cognitive deficit similar to what was observed in BRAF-mutated patients and in the general CFCS population. Conversely, congenital heart defects, particularly mitral valve and septal defects, and ocular anomalies seem to be less frequent among MEK1/MEK2 mutation-positive patients.     

Assessment of left ventricular volumes and function by cine-MR imaging depending on the investigator’s experience

AIMS: To analyze the reproducibility of LV volumes calculated by cardiac magnetic resonance imaging (CMRI) and to compare them to those obtained by conventional ventriculography. METHODS: A total of 30 patients with stable ischemic heart disease were prospectively included. Each underwent CMRI twice and ventriculography. Left ventricular end diastolic volume (EDV), end systolic volume (ESV) and LV ejection fraction (EF) were calculated by two radiologists at different level of experience. Intraobserver, interobserver and interstudy variabilities were assessed. RESULTS: The cut off values were: intraobserver variability (EDV, ESV, EF): 9.4 ml, 5.3 ml, 3.3% for well-trained radiologist; 13.1 ml, 7.5 ml, 4.1% for less-trained radiologist. interobserver variability: EDV: 11.7 and 10.4 ml; ESV: 7.0 and 6.6 ml; EF: 3.9 and 4.2%. interstudy variability (EDV, ESV, EF): 11.6 and 12.6 ml, 7.1 and 7.4 ml, 3.9 and 3.5%, for experienced and less-trained observers. Statistical differences were found between CMRI and ventriculography: CMRI underestimation of EDV and EF, overestimation of ESV. CONCLUSIONS: CMRI volumetric quantification of LV volumes and function is highly reproducible at different levels of experience, but not interchangeable with those obtained by ventriculography.     

Phenotypic detection of plasmid-acquired AmpC in Escherichia coli—evaluation of screening criteria and performance of two commercial methods for the phenotypic confirmation of AmpC production

The phenotypic detection of plasmid-acquired AmpC (pAmpC) in Escherichia coli is challenging, and molecular methods are required for confirmation. In addition to cefoxitin resistance, multiresistance and high-level resistance to cephalosporins have both been suggested as criteria for targeting isolates with pAmpC, but data to support these proposed criteria are lacking. A Swedish collection of 378 isolates with either putative chromosomal hyperproduction of AmpC (cAmpC) or pAmpC were subjected to disk diffusion and minimum inhibitory concentration (MIC) determination with the Etest. The frequency of resistance to gentamicin, ciprofloxacin, and trimethoprim among cAmpC and pAmpC was compared to elucidate the issue of multidrug resistance. Lastly, methods for the phenotypic confirmation of pAmpC were compared. One in-house disk diffusion method, one method employing NeoSensitabs (Rosco), and one Etest method (bioMérieux) were compared. The analysis of histograms based on both disk diffusion and the Etest showed that resistance [according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST)] to cefotaxime and/or ceftazidime occurred in almost all isolates. By coining resistance instead of non-susceptibility, the number of isolates required to subject to phenotypic testing/genotypic confirmation dropped by more than 40 %, without compromising the sensitivity substantially. Further, almost 70 % of isolates with pAmpC were non-multidrug resistant, clearly indicating that this is an inappropriate criterion for further investigation. The phenotypic tests all had more than 90 % sensitivity, and the best sensitivities were obtained with the in-house method and with the ceftazidime ± cloxacillin NeoSensitab. In conclusion, clinical resistance to cefotaxime and/or ceftazidime seems to be an appropriate criterion for pAmpC screening, and several phenotypic methods perform well for the phenotypic confirmation of AmpC production prior to genotypic confirmation.     

Inhibition of dermatophytes by the antimicrobial peptides human β-defensin-2, ribonuclease 7 and psoriasin

Previous studies have described some antibacterial effects of antimicrobial peptides (AMPs) expressed in human skin, but little is known about their possible activity against dermatophytes. Therefore we have tested the effects of human β-defensin 2 (hBD-2), ribonuclease 7 (RNase 7) and psoriasin on the in vitro growth of four dermatophyte species. Germinating conidia of Trichophyton rubrum, T. mentagrophytes, Microsporum canis and Epidermophyton floccosum were exposed in vitro to hBD-2, RNase 7, psoriasin and fluconazole. Subsequent fungal growth was measured photometrically over 168 hours. All AMPs significantly inhibited fungal growth, with the degree of inhibition dependent on the dermatophyte species and the specific AMP. E. floccosum was found to be the most susceptible species in that it was markedly suppressed by all AMPs, whereas M. canis was inhibited only by psoriasin. Overall, psoriasin was the most effective AMP and had even stronger inhibitory effects on some dermatophytes than fluconazole. Our findings show that AMPs expressed in human skin can, in principal, inhibit the growth of dermatophytes in vitro. Therefore the question whether AMPs are relevant for human protection against tineas is justified and should be addressed by investigating their role in vivo.