Hospital Environment as a Source of Azole-Resistant Strains with TR34/L98H and G448S Cyp51A Mutations.

Azole-resistant is an emerging worldwide problem with increasing reports of therapy failure cases produced by resistant isolates. A case of azole-resistant hospital colonization in a patient is reported here. Investigations of the hospital environment led to the recovery of strains harboring the TR34/L98H and the G448S Cyp51A azole resistance mechanisms.

MixInYeast: A Multicenter Study on Mixed Yeast Infections.

Invasive candidiasis remains one of the most prevalent systemic mycoses, and several studies have documented the presence of mixed yeast (MY) infections. Here, we describe the epidemiology, clinical, and microbiological characteristics of MY infections causing invasive candidiasis in a multicenter prospective study. Thirty-four centers from 14 countries participated.

Point Mutations in the 14-α Sterol Demethylase Cyp51A or Cyp51C Could Contribute to Azole Resistance in .

Infections caused by species are being increasingly reported. is the second most common species within this genus causing invasive infections in humans, and isolates showing azole resistance have been recently described. has three 51-related genes (51A, 51B, and 51C) encoding 14-α sterol demethylase-like enzymes which are the target of azole drugs.

Azole resistance survey on clinical Aspergillus fumigatus isolates in Spain.

Objectives: We aimed to assess the percentage of azole resistance in Aspergillus fumigatus in Spain.

Methods: Thirty participating Spanish hospitals stored all morphologically identified A. fumigatus sensu lato clinical isolates-regardless their clinical significance-from 15 February to 14 May 2019.

Fluconazole resistance is not a predictor of poor outcome in patients with cryptococcosis.

Background: Cryptococcus isolates with high MICs to fluconazole are increasingly reported, and a potential clinical impact has been advocated. However, there are different methods to evaluate fluconazole MICs and comparative analysis among such techniques and their comprehensive correlation with clinical outcome are not available.

Methods: Over a 13-year period (2000-2013), fluconazole MICs were determined for 62 cryptococcal isolates recovered from 22 patients with cryptococcosis using CLSI M27-A3, EUCAST, E test and Sensititre YeastOne, simultaneously.

Molecular Identification and Susceptibility Testing of Molds Isolated in a Prospective Surveillance of Triazole Resistance in Spain (FILPOP2 Study).

Antifungal resistance is increasing by the emergence of intrinsically resistant species and by the development of secondary resistance in susceptible species. A previous study performed in Spain revealed levels of azole resistance in molds of between 10 and 12.7%, but secondary resistance in was not detected.

Insight into the Significance of Aspergillus fumigatus cyp51A Polymorphisms.

Triazole antifungal compounds are the first treatment choice for invasive aspergillosis. However, in the last decade the rate of azole resistance among strains has increased notoriously. The main resistance mechanisms are well defined and mostly related to point mutations of the azole target, 14-α sterol demethylase (), with or without tandem repeat integrations in the promoter.

Could the determination of Aspergillus fumigatus mating type have prognostic value in invasive aspergillosis?

A clear link between mating type and virulence has been demonstrated for some fungal pathogens, but not for Aspergillus fumigatus as of yet. An association between mating type and invasiveness has recently been established. The mating type proportion (MAT1-1:MAT1-2) of 213 A.

Water Sources in a Zoological Park Harbor Genetically Diverse Strains of Clostridium Perfringens Type A with Decreased Susceptibility to Metronidazole.

The presence of Clostridium perfringens in water is generally regarded as an indicator of fecal contamination, and exposure to waterborne spores is considered a possible source of infection for animals. We assessed the presence and genetic diversity of C. perfringens in water sources in a zoological park located in Madrid (Spain).

A New Aspergillus fumigatus Typing Method Based on Hypervariable Tandem Repeats Located within Exons of Surface Protein Coding Genes (TRESP).

Aspergillus fumigatus is a saprotrophic mold fungus ubiquitously found in the environment and is the most common species causing invasive aspergillosis in immunocompromised individuals. For A. fumigatus genotyping, the short tandem repeat method (STRAf) is widely accepted as the first choice.

[Application of mass spectrometry in mycology].

MALDI-TOF (matrix-assisted laser desorption ionization time-of-flight) mass spectrometry (MS) is becoming an essential tool in most microbiology laboratories. At present, by using a characteristic fungal profile obtained from whole cells or through simple extraction protocols, MALDI-TOF MS allows the identification of pathogenic fungi with a high performance potential. This methodology decreases the laboratory turnaround time, optimizing the detection of mycoses.

Antifungal Susceptibility Testing of Ascomycetous Yeasts Isolated from Animals.

Recent studies suggest that antifungal resistance in yeast isolates of veterinary origin may be an underdiagnosed threat. We tested a collection of 92 ascomycetous yeast isolates that were obtained in Spain from birds, mammals and insects for antifungal susceptibility. MICs to amphotericin B and azoles were low, and no resistant isolates were detected.

Assessment of biofilm production in Candida isolates according to species and origin of infection.

The biofilm production (BP) of 200 clinical strains of Candida isolated during 2010-2013 were assessed using an in vitro model and a comparison of the results was made between species and between origins of the infections. The BP was assessed using the crystal violet assay, and the strains were classified as low, moderate, or high biofilm producers. Candida tropicalis had the highest values for BP, which varied depending on the origin of the infection.

Acquired multi-azole resistance in Candida tropicalis during persistent urinary tract infection in a dog.

Multi-azole resistance acquisition by Candida tropicalis after prolonged antifungal therapy in a dog with urinary candidiasis is reported. Pre- and post-azole treatment isolates were clonally related and had identical silent mutations in the ERG11 gene, but the latter displayed increased azole minimum inhibitory concentrations. A novel frameshift mutation in ERG3 was found in some isolates recovered after resistance development, so it appears unlikely that this mutation is responsible for multi-azole resistance.

Therapeutic drug monitoring of voriconazole helps to decrease the percentage of patients with off-target trough serum levels.

We monitored trough voriconazole serum concentrations from 107 patients (n = 258 samples) at 6 hospitals in Madrid. Most of the patients were male (67%) and had the following underlying conditions: hematological cancer (42%), solid organ transplantation (15%), chronic obstructive pulmonary disease (14%), human immunodeficiency virus infection (8.4%), solid cancer (5.

Genotyping and antifungal susceptibility testing of multiple Malassezia pachydermatis isolates from otitis and dermatitis cases in pets: is it really worth the effort?

A total of 216 colonies of Malassezia pachydermatis from 28 cases of fungal otitis or dermatitis in pets were genotyped by M13 fingerprinting and tested for antifungal susceptibility. A huge genetic diversity was found (157 M13 types in total), with all animals having a polyclonal pattern of infection (5.4 ± 1.

Sputum and bronchial secretion samples are equally useful as bronchoalveolar lavage samples for the diagnosis of invasive pulmonary aspergillosis in selected patients.

In the absence of histopathology studies of lung biopsies, the bronchoalveolar lavage (BAL) sample is preferred for the diagnosis of invasive pulmonary aspergillosis. Isolation of Aspergillus fumigatus from sputum and bronchial secretion samples are commonly interpreted as colonization or laboratory contamination, particularly in nonneutropenic patients. We studied if sputum/bronchial secretions and BAL samples are equally useful for the diagnosis of invasive pulmonary aspergillosis.

Aspergillus citrinoterreus, a new species of section Terrei isolated from samples of patients with nonhematological predisposing conditions.

The use of molecular identification techniques has revealed an increasing number of new species within Aspergillus section Terrei. We phenotyped a set of 26 clinical isolates that showed genetic differences from Aspergillus terreus sensu stricto by analyzing sequences from PCR-amplified β-tubulin and calmodulin genes and the internal transcribed spacer region. Since the isolates were phylogenetically and morphologically different from all of the members of Aspergillus section Terrei, they are described here as a new species, Aspergillus citrinoterreus, so named because it produces a diffusible yellowish pigment in agar.

Microsatellite (STRAf) genotyping cannot differentiate between invasive and colonizing Aspergillus fumigatus isolates.

We studied whether short tandem repeats of Aspergillus fumigatus (STRAf) can differentiate between invasive and colonizing genotypes of A. fumigatus. Of the 395 genotypes detected (n = 1,373 isolates), 50 were clusters and 24 (6% of all genotypes) involved the patients with invasive aspergillosis and those colonized with A.

Effect of melatonin and analogues on corneal wound healing: involvement of Mt2 melatonin receptor.

Purpose: We have investigated the effect of melatonin and its analogues on rabbit corneal epithelial wound healing.

Methods: New Zealand rabbits were anaesthetised and wounds were made by placing Whatman paper discs soaked in n-heptanol on the cornea. Melatonin and analogues (all 10 nmol) were instilled.

Is it feasible to diagnose catheter-related candidemia without catheter withdrawal?

Many bloodstream infections (BSI) in patients with central venous catheters (CVC) are not catheter-related (CR). Assessment of catheter involvement without catheter withdrawal has not been studied in candidemia. We assessed the value of conservative techniques to evaluate catheters as the origin of candidemia in patients with CVC in a prospective cohort study (superficial Gram stain and culture, Kite technique (Gram stain and culture of the first 1 cm blood drawn from the CVC), proportion of positive blood cultures (PPBCs), differential time to positivity (DTP), and minimal time to positivity (MTP)).

Is biofilm production a predictor of catheter-related candidemia?

Catheter-related candidemia (CRC) is typically a biofilm related disease, but it is mostly unknown if the production of biofilm is a feature exclusively shown by Candida spp. isolates causing CRC. We performed an in vitro biofilm assay using Candida isolates obtained from the blood of patients with candidemia.