Fungal diversity and surfactant-producing fungi in oil contaminated environments.

AIMS: To investigate fungal diversity and biosurfactant-producing fungi in four oil-contaminated sites. METHODS AND RESULTS: Water and sediment samples were collected from four sites in Brittany (France), over two periods, in winter/spring and summer. Fungal diversity was investigated using a metagenetic approach targeting the ITS2 region. Surface-active compound production of 701 fungal isolates collected from these samples after direct plating or following enrichment was assessed using oil spreading and Parafilm M tests. Fungal communities were highly diverse and the main dominant fungal taxa were members of the Cladosporium, Penicillium, Pseudeurotium, Phoma, Aspergillus, and Trichoderma as well as Ochroconis, Fusicolla, and Aureobasidium genera in specific sites. A total of 179 isolates (25.5% of total isolates) were positive to at least one of the screening tests, while 105 were positive to both tests. Major genera among the positive isolates were Fusarium, Trichoderma, Candida, and Penicillium. Six isolates belonging to Aureobasidium pullulans, Mucor griseocyanus, Trichoderma citrinoviride, Trichoderma harzianum, Trichodermalongibrachiatum, and Diaporthe eres showed promising activities. CONCLUSIONS: The present study highlighted the fungal diversity of oil-contaminated environments and the fact that surface-active compound production is widespread in fungi originating from these habitats.

Use of metabarcoding and source tracking to identify desirable or spoilage autochthonous microorganism sources during black olive fermentations.

This study aimed at investigating the influence of the process environment and raw materials as sources of microorganisms during Nyons black table olive fermentations. Fermented olives and/or brine from spoiled fermentation tanks were analyzed and compared to good quality samples from fermentations collected during 3 consecutive harvest years. Fresh olives, salt and different process environment samples were also analyzed. Microbial diversity of all samples was analyzed using 16S and ITS2 amplicon sequencing and SourceTracker tool was used to investigate links between environment, raw materials and fermentation samples. First, comparison of microbial diversity in control and most spoiled fermentations revealed striking differences in bacterial composition with an overall higher diversity in spoiled fermentations especially for lactic acid bacteria with Lentilactobacillus buchneri, Lentilactobacillus parafarraginis dominating in brine and Pediococcus parvulus, Pediococcus ethanolidurans dominating in olive fruits. Fungal communities were similar in composition although higher abundances of Pichia membranifaciens and Penicillium carneum/roqueforti were observed in spoiled samples. Secondly, process environment samples were characterized by high bacterial and fungal diversity, especially compared to fresh olive fruits. Overall, dominant fungal species in control fermentations were also found in most environmental samples revealing a "house mycobiota". SourceTracker analysis further highlighted the contribution of brine and water from the optical sorter as a source of fungi. Most interestingly, spoilage fungi and most bacteria were retrieved in brine and environmental samples while others such as P. ethanolidurans were only found in environmental samples indicating that the studied spoilage originated from a fermentation deviation rather than a punctual contamination. Taken altogether, these results highlighted the positive and negative influence of the process environment and emphasized the relevance of studying it to better understand microbial vectors occurring during food fermentations, especially natural ones.

Development and application of MALDI-TOF MS for identification of food spoilage fungi.

Filamentous fungi are frequently involved in food spoilage and cause important food losses and substantial economic damage. Their rapid and accurate identification is a key step to better manage food safety and quality. In recent years, MALDI-TOF MS has emerged as a powerful tool to identify microorganisms and has successfully been applied to the identification of filamentous fungi especially in the clinical context. The aim of this study was to implement a spectral database representative of food spoilage molds. To this end, after application of a standardized extraction protocol, 6477 spectra were acquired from 618 fungal strains belonging to 136 species and integrated in the VITEK MS database. The performances of this database were then evaluated by cross-validation and ∼95% of correct identification to the species level was achieved, independently of the cultivation medium and incubation time. The database was also challenged with external isolates belonging to 52 species claimed in the database and 90% were correctly identified to the species level. To our best knowledge, this is the most comprehensive database of food-relevant filamentous fungi developed to date. This study demonstrates that MALDI-TOF MS could be an alternative to conventional techniques for the rapid and reliable identification of spoilage fungi in food and industrial environments.