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Personne :
Lessard, Marie-Hélène

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Lessard

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Marie-Hélène

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Université Laval. Département des sciences des aliments et de nutrition

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ncf11856500

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Résultats de recherche

Voici les éléments 1 - 7 sur 7
  • PublicationAccès libre
    Le suivi de la croissance et de l'activité spécifique des mycètes pendant l'affinage du Camembert
    (2014) Lessard, Marie-Hélène; Labrie, Steve; St-Gelais, Daniel
    Le suivi de la croissance et de l’activité de la microflore fongique de surface des fromages de type Camembert demeure un défi actuel. Cet écosystème est principalement composé de Geotrichum candidum et de Penicillium camemberti. Ces mycètes sont responsables de l’apparence et du développement des caractéristiques sensorielles pendant la période d’affinage des fromages à croûte fleurie. Ils ont donc un intérêt technologique pour l’industrie fromagère. Malgré leur importance, peu de travaux ont porté sur l’étude approfondie de l’écosystème de surface des fromages à croûte fleurie en cours d’affinage. Étant donné la nature filamenteuse, envahissante et multicellulaire des mycètes, les méthodes traditionnelles de dénombrement sur milieu gélosé sont imprécises. Pour y remédier, une méthode de quantification par PCR en temps réel (qPCR) a permis de suivre une population mixte contenant les mycètes les plus communs (Penicillium camemberti, Geotrichum candidum, Debaryomyces hansenii et Kluyveromyces lactis) sur des Camembert‐modèles, en fonction du temps d’affinage. En général, P. camemberti et G. candidum dominent l'écosystème et K. lactis demeure peu abondant. Lorsque D. hansenii est présent, il inhibe la croissance de K. lactis en plus de réduire celle des autres mycètes. Les informations aujourd’hui disponibles concernant ces mycètes sont principalement d’ordre biochimique et les gènes portés par G. candidum et par P. camemberti, de même que leurs profils d’expression pendant l’affinage sont peu décrits. La première étude métatranscriptomique de l’écosystème fongique du fromage Camembert a donc été réalisée et une base de données appelée CamemBank01 contenant près de 8000 gènes a pu être générée. L’annotation fonctionnelle des séquences obtenues a révélé que les fonctions prédominantes retrouvées dans CamemBank01 étaient associées au développement des propriétés sensorielles et que ces fonctions étaient principalement exprimées dans les deux premières semaines de l’affinage. Les résultats présentés dans cette thèse suggèrent que plusieurs gènes exprimés pendant l’affinage du Camembert pourraient servir de biomarqueurs. Les travaux futurs amèneront une meilleure compréhension de l’activité de ces champignons et permettront une utilisation optimale des ferments d’affinage.
  • PublicationAccès libre
    Quantitative PCR reveals the frequency and distribution of 3 indigenous yeast species across a range of specialty cheeses
    (American Dairy Science Association, 2022-09-14) Lamarche, Andréanne; Lessard, Marie-Hélène; Viel, Catherine; Turgeon, Sylvie; St-Gelais, Daniel; Labrie, Steve
    Indigenous microorganisms are important components of the complex ecosystem of many dairy foods including cheeses, and they are potential contributors to the development of a specific cheese's sensory properties. Among these indigenous microorganisms are the yeasts Cyberlindnera jadinii, Pichia kudriavzevii, and Kazachstania servazzii, which were previously detected using traditional microbiological methods in both raw milk and some artisanal specialty cheeses produced in the province of Québec, Canada. However, their levels across different cheese varieties are unknown. A highly specific and sensitive real-time quantitative PCR assay was developed to quantitate these yeast species in a variety of specialty cheeses (bloomy-rind, washed-rind, and natural-rind cheeses from raw, thermized, and pasteurized milks). The specificity of the quantitative PCR assay was validated, and it showed no cross-amplification with 11 other fungal microorganisms usually found in bloomy-rind and washed-rind cheeses. Cyberlindnera jadinii and P. kudriavzevii were found in the majority of the cheeses analyzed (25 of 29 and 24 of 29 cheeses, respectively) in concentrations up to 104 to 108 gene copies/g in the cheese cores, which are considered oxygen-poor environments, and 101 to 104 gene copies/cm2 in the rind. However, their high abundance was not observed in the same samples. Whereas C. jadinii was present and dominant in all core and rind samples, P. kudriavzevii was mostly present in cheese cores. In contrast, K. servazzii was present in the rinds of only 2 cheeses, in concentrations ranging from 101 to 103 gene copies/cm2, and in 1 cheese core at 105 gene copies/g. Thus, in the ecosystems of specialty cheeses, indigenous yeasts are highly frequent but variable, with certain species selectively present in specific varieties. These results shed light on some indigenous yeasts that establish during the ripening of specialty cheeses.
  • PublicationRestreint
    A sequencing approach targeting the 16S rRNA gene unravels the biofilm composition of spiral-wound membranes used in the dairy industry
    (Springer, 2016-10-17) Chamberland, Julien; Lessard, Marie-Hélène; Doyen, Alain; Labrie, Steve; Pouliot, Yves
    Few data are available concerning the composition of biofilms found at the surface of filtration membranes, which, to some extent, explains the long-term failure of numerous strategies developed to control biofouling. This preliminary study intended to design a metagenomic tool targeting the 16S rRNA gene in order to unravel a general portrait of bacterial communities found on spiral-wound membranes used in the dairy industry. A total of seven spiral-wound membrane elements (ultrafiltration, nanofiltration, or reverse osmosis) at the end of their useful lifetimes were collected from different dairy plants. Targeted analysis of the 16S rRNA genes of the metagenome extracted from the membranes revealed their bacterial diversity via high-throughput sequencing technology (Miseq, Illumina). It was found that the nature of the filtered fluid (milk, whey, water) explained 58.6 % of the variance observed between communities found on membranes. Treatments applied on dairy fluids (milk pasteurization, whey bleaching or whey ultrafiltration) induced a selective pressure that affected the diversity of bacterial communities found on membranes and the proportions of spore-former bacteria among them. This work provides the first complete bacterial portrait of the biofilm composition of spiral-wound membranes used in the dairy industry. It suggests that the nature of the filtered fluid and potentially filtration operating parameters may be important elements to consider in order to design new cleaning strategies or preventive measures targeting biofouling.
  • PublicationAccès libre
    Découvrir les variations génomiques entre des souches de Lactococcus lactis ssp. cremoris par hybridation suppressive soustractive et par analyse de séquences multi-locus
    (2009) Lessard, Marie-Hélène; Roy, Denis; LaPointe, Gisèle
    La diversité génomique de Lactococcus lactis a été évaluée par Hybridation Suppressive Soustractive (SSH) et par ±Multilocus Sequence Type Analysis¿ (MLSA). Les gènes présentant la plus grande diversité sont les gènes de prophages, de réplication plasmidique et les transposases. De plus, pour les souches IL1403, MG1363, SKll et A TCC-19257, des transporteurs (sucre, peptides, acides aminés), des peptidases et des glycosyltransférases spécifiques ont été isolées. Les résultats pour les 21 loci partiellement séquencés pour les souches SK11 , A TCC-I9257, HP, Wg2 et ES ont permis de construire un schéma MLSA optimal avec six gènes, selon le degré de polymorphisme, l'origine des loci et le nombre d'allèles par loci. La similarité des séquences démontre la relation phylogénétique entre les souches, qui peuvent être divisées en deux groupes centrés autour de la souche SK11 et A TCC 19257. Ce projet de recherche a permis de repérer des différences spécifiques entre les souches de Lactococcus lactis tout en présentant l'utilisation d'outils de génomique comparative pouvant être appliqués au domaine alimentaire.
  • PublicationRestreint
    Biofouling of ultrafiltration membrane by dairy fluids : characterization of pioneer colonizer bacteria using a DNA metabarcoding approach
    (Elsevier, 2017-01-25) Chamberland, Julien; Lessard, Marie-Hélène; Doyen, Alain; Labrie, Steve; Pouliot, Yves
    Biofouling of filtration membranes is a major quality and performance issue for the dairy industry. Because biofilms that survive cleaning cycles become resistant over time, prevention strategies limiting the adhesion of bacteria to membranes should be prioritized for sustainable control of biofouling. However, this cannot be achieved because the pioneer bacteria colonizing these membranes are still unknown. Consequently, the objective of this study was to characterize pioneer bacteria on the filtration membrane surface and to measure the effect of filtration operational parameters on their diversity. Thus, milk and cheese whey were filtered for 5 h in concentration mode at 10 and 40°C using a laboratory-scale crossflow filtration system equipped with flat-sheet ultrafiltration membranes. Pioneer colonizer bacteria found on membranes after a chlorinated alkaline cleaning cycle were identified using a metabarcoding approach targeting the 16S ribosomal RNA genes. Our results suggested that prevention strategies targeting biofouling should consider the nature of the filtered fluid and the feed temperature (36.15 and 5.09% of the variances observed on membranes, respectively), as well as the microbial environment of the dairy processing plant. In the future, it is hypothesized that cleaning prevention strategies will be specific to each dairy processor and their operational parameters.
  • PublicationAccès libre
    The composition of Camembert cheese-ripening cultures modulates both mycelial growth and appearance
    (American Society of Microbiology, 2012-01-13) Lessard, Marie-Hélène; Bélanger, Gaétan; Labrie, Steve; St-Gelais, Daniel
    The fungal microbiota of bloomy-rind cheeses, such as Camembert, forms a complex ecosystem that has not been well studied, and its monitoring during the ripening period remains a challenge. One limitation of enumerating yeasts and molds on traditional agar media is that hyphae are multicellular structures, and colonies on a petri dish rarely develop from single cells. In addition, fungi tend to rapidly invade agar surfaces, covering small yeast colonies and resulting in an underestimation of their number. In this study, we developed a real-time quantitative PCR (qPCR) method using TaqMan probes to quantify a mixed fungal community containing the most common dairy yeasts and molds: Penicillium camemberti, Geotrichum candidum, Debaryomyces hansenii, and Kluyveromyces lactis on soft-cheese model curds (SCMC). The qPCR method was optimized and validated on pure cultures and used to evaluate the growth dynamics of a ripening culture containing P. camemberti, G. candidum, and K. lactis on the surface of the SCMC during a 31-day ripening period. The results showed that P. camemberti and G. candidum quickly dominated the ecosystem, while K. lactis remained less abundant. When added to this ecosystem, D. hansenii completely inhibited the growth of K. lactis in addition to reducing the growth of the other fungi. This result was confirmed by the decrease in the mycelium biomass on SCMC. This study compares culture-dependent and qPCR methods to successfully quantify complex fungal microbiota on a model curd simulating Camembert-type cheese.
  • PublicationAccès libre
    Metatranscriptome analysis of fungal strains Penicillium and Geotrichum reveal cheese matrix breakdown and potential development of sensory properties of ripened Camembert-type cheese
    (MEDLINE/PubMed (U.S. National Library of Medicine), 2014-03-26) Viel, Catherine; Lessard, Marie-Hélène; Boyle, Brian; Labrie, Steve; St-Gelais, Daniel
    Camembert-type cheese ripening is driven mainly by fungal microflora including Geotrichum candidum and Penicillium camemberti. These species are major contributors to the texture and flavour of typical bloomy rind cheeses. Biochemical studies showed that G. candidum reduces bitterness, enhances sulphur flavors through amino acid catabolism and has an impact on rind texture, firmness and thickness, while P. camemberti is responsible for the white and bloomy aspect of the rind, and produces enzymes involved in proteolysis and lipolysis activities. However, very little is known about the genetic determinants that code for these activities and their expression profile over time during the ripening process.