The World Mycotoxin Forum

10.11.- 12.11.2014
Vienna, Austria
http://www.bastiaanse-communication.com/wmf/wmf.html

 

Speed presentation

We are looking forward to welcoming you to the 6 minute speed presentation of Emmanuel RONDAGS about the following poster, providing an overview of their research and inspiring the audience to visit their poster.

Tuesday, 11. November 2014

16:15 - 17:00 h, Session 8

 

"Fusarium species fast identification and associated mycotoxins detection by Maldi-TOF"

1Julien Billard, 1Sophie Heit, 2Philippa Hart, 3Cedric Paris, 4Renaud Ioos, 2Omar Belgacem, 1Xavier Framboisier and 1Emmanuel Rondags

 

1 Laboratoire Réactions et Génie des Procédés, Université de Lorraine, Vandoeuvre, France

2 Kratos analytical, Manchester, United Kingdom

3 ENSAIA, Laboratoire commun d’analyses, Vandoeuvre-lès-Nancy, France

4 ANSES, Laboratoire de la Santé des Végétaux - Unité de Mycologie, Malzéville, France 

 

Summary

The barley-Malt-Beer chain is regularly affected by Fusarium contaminations leading to reduced yields, technological transformations concerns as well as safety issues. Due to the difficulty to predict the nature and levels of both mycological and mycotoxins contaminations in an evolving environment, a proper hygienic control of the whole chain is hard to achieve. Fast corrective actions based on phytosanitary and/or biological treatments as well as technological practices modifications are now required.

In order to speed up the accurate adaptation of treatments and production processes, fast biological and mycotoxins analytical techniques are needed. In this context, the use of Maldi-TOF for the fast identification of Fusarium species encountered in the Barley-Malt-Beer chain and for the detection of some associated mycotoxins is evaluated.

As far as identification and detection are concerned, the adaptation of the classical Maldi-TOF bacterial identification protocols for the particular case of moulds, with a focus on typical Fusarium species affecting Barley and Malt, will be initially described. The ability of this technique to detect mycotoxins will then be addressed mainly on T2 toxin, either in solution or associated with biomass in order to display the Maldi-TOF ability to both identify Fusarium and evaluate the associated toxinogenic potential.

At last, the ability of this technique to cope with defined mixed Fusarium cultures will be examined. These promising results however highlight the need for material and software evolutions that will be discussed in relation with a global strategy for improved mycological control of cultures.

 


 

Poster presentation 

"Adaptation of bacteriological protocols for the fast fungal identification and associated mycotoxins detection by Maldi-TOF"

 

1Julien Billard, 1Sophie Heit, 2Philippa Hart, 3Cedric Paris, 4Renaud Ioos, 2Omar Belgacem, 1Xavier Framboisier and 1Emmanuel Rondags

1 Laboratoire Réactions et Génie des Procédés, Université de Lorraine, Vandoeuvre, France

2 Kratos analytical, Manchester, United Kingdom

3 ENSAIA, Laboratoire commun d’analyses, Vandoeuvre-lès-Nancy, France

4 ANSES, Laboratoire de la Santé des Végétaux - Unité de Mycologie, Malzéville, France

 

Summary

In a global context of rising mycological and associated mycotoxins concerns in the agricultural chains and especially in the barley-Malt-Beer chain, the use of Maldi-TOF for the fast phenotypic mould identification and mycotoxins detection is here evaluated. This short presentation will consequently focus on the adaptation of existing material and software initially developed for bacterial identification to moulds related purposes.

Due to the typical concerns of the chain, this study focused on graminearum (generic cereal contamination), langsethiaepoae (T2 and HT2) and tricinctum (Enniatins) species of the Fusarium genus. These strains were isolated and identified by either IFBM or ANSES from field samples. Strains from the different species were cultivated in both liquid and solid media, in pure and mixed cultures for further Maldi-TOF analyses. A first step of this work consisted in the adaptation of the sample preparation conditions in order to get proper analytical signal as moulds are generally far more receptive to laser ionization than bacteria, due to different parietal composition. Then, pure culture samples have been analyzed and the corresponding spectra treated to fill the Saramis database. 

Our results show that species level Fusarium identification is possible with the considered strains, even if the number of mass peaks is actually lower than the one obtained with bacteria. This identification can be achieved on at least 3 days liquid or 5 days solid cultures, with an analysis time of only some minutes, which speeds up the identification protocol. As far as mycotoxins detection is concerned, both enniatins, and T2 and HT2 toxins were accurately detected with the producing strains, whereas no detection took place with non-producing strains. 

However, mycotoxins detection was only achieved with solid media samples, which seems logical as solid state cultures correspond to the mycotoxin production physiological state. At last, our work showed that, without Saramis software adaptation, co-identification of Fusarium species in mixed cultures with up to 3 different species was achievable.

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