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 Aspergillus fumigatus was not detected. We used itraconazole (ITZ)-supplemented medium to select resistant strains. A total of 500 plates supplemented with 2 mg/liter of ITZ were sent to 10 Spanish tertiary hospitals, and molecular identification and antifungal susceptibility testing were performed. In addition, the cyp51A gene in those A. fumigatus strains showing azole resistance was sequenced. A total of 493 isolates were included in the study. Sixteen strains were isolated from patients with an infection classified as proven, 104 were isolated from patients with an infection classified as probable, and 373 were isolated from patients with an infection classified as colonization. Aspergillus was the most frequent genus isolated, at 80.3%, followed by Scedosporium-Lomentospora (7.9%), Penicillium-Talaromyces (4.5%), Fusarium (2.6%), and the order Mucorales (1%). Antifungal resistance was detected in Scedosporium-Lomentospora species, Fusarium, Talaromyces, and Mucorales Three strains of A. fumigatus sensu stricto were resistant to azoles; two of them harbored the TR34+L98H mechanism of resistance, and the other one had no mutations in cyp51A The level of azole resistance in A. fumigatus remains low, but cryptic species represent over 10% of the isolates and have a broader but overall higher range of antifungal resistance.

Invasive scedosporiosis in lung transplant recipients: A nine-year retrospective study in a tertiary care hospital.

BACKGROUND: Scedosporium species and Lomentospora prolificans (Sc/Lp) are emerging molds that cause invasive disease associated with a high mortality rate. After Aspergillus, these molds are the second filamentous fungi recovered in lung transplant (LT) recipients. AIMS: Our objective was to evaluate the incidence, risk factors and outcome of Sc/Lp infections in LT recipients at a tertiary care hospital with a national reference LT program. METHODS: A nine-year retrospective study was conducted. RESULTS: During this period, 395 LT were performed. Positive cultures for Sc/Lp were obtained from twenty-one LT recipients. Twelve patients (incidence 3.04%) developed invasive scedosporiosis (IS). In 66.7% of the patients with IS the invasive infection was defined as a breakthrough one. The main sites of infection were lungs and paranasal sinuses. Most of the patients received combination antifungal therapy. The IS crude mortality rate after 30 days was 16.7%, and 33.3% after a year. CONCLUSIONS: Our study highlights improved survival rates associated with combination antifungal therapy in LT recipients and underlines the risk of breakthrough infections in patients with allograft dysfunction on nebulized lipidic amphotericin B prophylaxis. In addition to pretransplant colonization, acute or chronic organ dysfunctions seem to be the main risk factors for IS.

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. Isolates showing azole resistance according to the EUCAST 9.3.2 methodology were molecularly identified and the cyp51A gene was studied in A. fumigatus sensu stricto isolates. RESULTS: Eight hundred and forty-seven isolates from 725 patients were collected in 29 hospitals (A. fumigatus sensu stricto (n = 828) and cryptic species (n = 19)). Isolates were mostly from the lower respiratory tract (94.0%; 797/847). Only cryptic species were amphotericin B resistant. Sixty-three (7.4%) out of the 847 isolates were resistant to ≥1 azole(s). Azole resistance was higher in cryptic species than in A. fumigatus sensu stricto (95%, 18/19 vs. 5.5%, 45/828); isavuconazole was associated to the lowest number of non-wild type isolates. The dominant mechanism of resistance was the presence of TR34-L98H substitutions (n = 24 out of 63). Out of the 725 patients, 48 (6.6%) carried either cryptic species (n = 14) or A. fumigatus sensu stricto (n = 34; 4.7%) resistant isolates. Aspergillus fumigatus sensu stricto harbouring either the TR34-L98H (n = 19) or TR46/Y121F/T289A (n = 1) mutations were detected in patients in hospitals located at 7/24 studied cities. DISCUSSION: Of the patients, 6.6% carry azole-resistant A. fumigatus sensu lato isolates in Spain. TR34-L98H is the dominant cyp51A gene substitutions, although its presence is not widespread.

Influence of environmental conditions and pollution on the incidence of Streptococcus pneumoniae infections.

Fossil fuel derived pollutants (SO2, NO), dry air and cold increase the incidence of S. pneumoniae infections http://ow.ly/RnLW30gogb1.

Update on the diagnosis of invasive fungal infection / Actualización en el diagnóstico de la infección fúngica invasora

The number of patients at risk of suffering invasive fungal infection (IFI) is increasing. Because of its high mortality, new rapid and accurate diagnostic tools are needed. Last advances in invasive candidiasis diagnosis comprise Peptide Nucleic Acid Fluorescent In-Situ Hybridization (PNA-FISH), direct MALDI-TOF or multiplex acid nucleic testing. While all of them rely in positive blood cultures, T2Candida(R) uses PCR coupled with T2Magnetic resonance detection directly in whole blood, allowing detection of 1-3 UFC/mL of Candida in about four hours. Beyond galactomannan (GM), novelties in IFI caused by molds include the international standardization of PCR techniques, with several commercial kits available. A combination of GM and PCR appears to be a good diagnostic strategy for invasive aspergillosis. PCR coupled to electrospray ionization/mass spectrometry and detection of volatile organic compounds in exhaled air by gas chromatography/mass spectrometry are other promising approaches to IFI diagnostic that still need to be validated (AU)

El número de pacientes en riesgo de padecer infección fúngica invasora (IFI) está en aumento. Debido a su elevada mortalidad, es necesario disponer de nuevas herramientas diagnósticas más rápidas, sensibles y específicas que las que disponemos en la actualidad. Los últimos avances en el diagnóstico de la candidiasis invasora incluyen Hibridación In Situ de Ácidos Péptidonucleicos (PNA-FISH), MALDI-TOF directo o PCR múltiple. Mientras que todas estas técnicas se realizan sobre frascos de hemocultivo positivos, T2Candida(R) se basa en una PCR con detección por resonancia magnética T2 directamente en sangre total, y permite la detección de entre 1-3 UFC/mL de Candida en aproximadamente 4 horas. Más allá del galactomanano (GM), una de las últimas novedades en el diagnóstico de IFI causada por hongos filamentosos es la estandarización internacional de las técnicas moleculares, con la aparición de varios kits comerciales. Una buena estrategia para el diagnóstico de aspergilosis invasora es la combinación de GM y PCR. La PCR asociada a ionización por electrospray/espectrometría de masas y la detección de compuestos orgánicos volátiles en aire exhalado mediante cromatografía de gases asociada a espectrometría de masas son otras aproximaciones prometedoras al diagnóstico de IFI que aún deben ser validadas (AU)