PREVALENCE, GEOGRAPHIC RISK FACTORS, AND DEVELOPMENT OF A STANDARDIZED PROTOCOL FOR FUNGAL ISOLATION IN CYSTIC FIBROSIS: RESULTS FROM THE INTERNATIONAL PROSPECTIVE STUDY “MFIP”

Authors:

Laurence Delhaes1,2, Kada Touati1, Odile Faure-Cognet3, Muriel Cornet3, Françoise Botterel4, Eric DannaouiI5, Florent Morio6, Patrice Lepape6, Fréderic Grenouillet7, Loic Favennec8, Solène Le Gal9, Gilles Nevez9, Alain Duhamel1, Andrew Borman10, Veroniek Saegeman11, Katrien Lagrou11, Elia Gomez12, Maiz-Luis Carro12, Rafael Canton12, Silvia Campana13, Walter Buzina14, Sharon Chen15, Wieland Meyer15, Emmanuel Roilides16, Maria Simitsopoulou16, Esther Manso17, Lisa Cariani18, Arianna Biffi18, Ersilia Fiscarelli19, Gabriella Ricciotti19, Marc Pihet20, Jean-Philippe Bouchara20

Affiliations:

1University & CHU of Lille, F-59000 Lille, France;

2University & CHU of Bordeaux, INSERM 1045, FHU ACRONIM, France. (current address), France;

3Univ. Grenoble Alpes, CNRS, Grenoble INP, CHU Grenoble Alpes, TIMC-IMAG, Grenoble, France e;

4Dynamyc Team, University of Paris Est Créteil, France;

5University of Paris-5, France;

6University of Nantes, France;

7Besançon University Hospital, France;

8University of Rouen, France;

9University of Brest, France;

10PHE Mycology Reference Laboratory, Bristol, UK;

11University of Leuven, and National Reference center for Mycosis, Belgium;

12Hosital Universitario Ramón y Cajal and Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS) University of Madrid, Spain;

13University of Florence, Italia;

14Medical university of Graz, Austria;

15Molecular Mycology Research Laboratory, Marie Bashir Institute for Biosecurity and Emerging infections, University of Sydney, Australia;

16University of Thessaloniki, Greece;

17Ancona Hospital, Italia;

18Microbiology and Cystic Fibrosis Microbiology laboratory, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico Milano, Italia;

19Bambino Children’s Hospital Rome, Italia;

20University of Angers, France

 

What was your research question?

Beside bacteria, there are many other microorganisms in the chest of cystic fibrosis (CF) patients including fungi, which recent research has been shown to be harmful. To better understand the role of such fungi, we conducted an international study named MFIP for “MucoFong International Project”.

Why is this important?

At the moment, different laboratories are using different methods to detect these fungi, making it difficult to determine the full picture of the impact of fungal infections on CF, and limiting the ability to improve patient management. To address this issue, MFIP has been undertaken with the aims to: (i) determine how frequently harmful fungi are isolated from the CF patient sputa (mucus), (ii) compare how well laboratory methods perform in the detection of these fungi, and (iii) standardize a universal laboratory method for the routine detection and identification of these fungi in diagnostic laboratories supporting CF units around the world.

What did you do?

An international study was set up (https://www.ecfs.eu/special-projects/mucofong-international-project) in which all participating centers used the same method, approved by the Lille Ethical Committee (Observational study 2012-042). 469 sputa were collected from CF patients followed-up in 18 European and one Australian CF center. Sputa from CF patients were added onto eight different growth media and inspected for the growth of fungi at least twice weekly during the 15 days incubation (Figure1). By statistical analyses of the obtained results the best way to grow fungi from sputa of CF patients was determined, and resulted in recommendations.

What did you find?

Fungi were detected more often from sputa of older patients. Fungi are divided into molds and yeasts. Amongst molds, a fungus called Aspergillus fumigatus was most frequently isolated (34.5% of sputa), followed by another mold: Scedosporium (5.1%). The most frequently isolated yeast was Candida albicans (47.7%). We identified a growing European North-to-South difference relating to how often Scedosporium was isolated, while yeasts, Aspergillus and other molds were more common in the Northern regions.

Regarding recommendations, CF laboratories should employ 2 media, called Yeast-Peptone-Dextrose-Agar or Sabouraud, and B(+), which should be supplemented with another medium when looking for Scedosporium or Exophiala.

What does this mean and reasons for caution?

Results from this MFIP study have generated a better understanding of what fungi are present in the lungs of CF patients, highlighting age differences as well as geographical differences amongst fungi found in sputa of CF patients. The study reinforces the importance of regular checking of sputa from CF patients for the presence of fungi, even if they feel well, as recommended by most current clinical guidelines.

What’s next?

Using the new methods recommended here will help to work out how lung function is impacted by the presence of fungi, and if certain fungi are worse than others.

 

 

Figure 1: The appearance of Scedosporium apiospermum (A) and Aspergillus fumigatus (B) on Sabouraud growth medium.



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