Personne : Gendron, Louis
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Gendron
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Louis
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Université Laval. Faculté des sciences et de génie
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ncf11907401
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Publication Accès libre The use of fluorescent bacteriophages to study viroaerosol characteristics(2014) Gendron, Louis; Duchaine, Caroline; Côté, DanielPour bien comprendre et contrôler les aérosols contenant des virus (viroaérosol), un modèle de laboratoire approprié est requis. Pour cette étude, trois bactériophages : P008 couplé au SYBR Gold, PP01 exprimant la GFP et ʎ exprimant la EYFP ont été comparés entre eux et à des microsphères fluorescentes non-biologiques pour leur potentiel en tant que modèle de laboratoire en aérovirologie. Les modèles viraux ont été aérosolisés à partir d’un tampon de phage en utilisant un nébuliseur de TSI (modèle 9301) connecté à une chambre d’aérosols. La taille aérodynamique des aérosols ainsi que leur distribution ont été déterminées à l’aide d’un spectromètre de particules aérodynamique (APS, TSI modèle 3321). Les échantillons de viroaérosols ont été capturés à l’aide d’un impacteur Andersen à six étages contenant soit du tampon de phage à l’intérieur des plaques de chaque étage ou un milieu solide (agar à 1.5%). Les techniques des plages de lyse, du qPCR et la microscopie à fluorescence ont été utilisées pour quantifier les virus récupérés sur les étages de l’impacteur. La microscopie à fluorescence a aussi été utilisée pour quantifier et analyser les modèles viraux sur des particules seules et sur milieu solide. L’ADN viral, des plages de lyse ainsi que des particules fluorescentes ont été observées sur les étages 3 à 6 de l’échantillonneur ce qui corrélait avec les données obtenues par l’APS. La microscopie à fluorescence a permis de visualiser les modèles viraux sur ou à l’intérieur des particules d’aérosols. Ces résultats confirment que les virus peuvent être présents dans l’atmosphère sous forme d’aérosol dont la dimension est bien plus grande que celle de leur propre taille, et que les virus en aérosol peuvent être quantifiés et observés en utilisant la microscopie à fluorescence. L’ensemble de ces résultats suggèrent qu’un bactériophage fluorescent serait un excellent modèle de laboratoire pour étudier le comportement des virus dans l’air.Publication Restreint Evaluation of bacterial contaminants found on unused paper towels and possible postcontamination after handwashing : a pilot study(Mosby Co., 2011-12-19) Duchaine, Caroline; Gendron, Louis; Moineau, Sylvain; Trudel, LucBackground: Bacterial contamination is a concern in the pulp and paper industry. Not only is the machinery contaminated but also can be the end-paper products. Bacterial transmission from unused paper towels to hands and surfaces is not well documented. Methods: The culturable bacterial community of 6 different unused paper towel brands was determined by culture methods and by sequencing the 16S ribosomal DNA of bacterial contaminants. Next, we investigated the possible airborne and direct contact transmissions of these bacterial contaminants during hand drying after washing. Results: Between 10(2) and 10(5) colony-forming units per gram of unused paper towels were isolated from the different paper towel brands. Bacteria belonging to the Bacillus genus were by far the most abundant microorganisms found (83.0%), followed by Paenibacillus (15.6%), Exiguobacterium (1.6%), and Clostridium (0.01%). Paper towels made from recycled fibers harbored between 100- to 1,000-fold more bacteria than the virgin wood pulp brand. Bacteria were easily transferred to disposable nitrile gloves when drying hands with paper towels. However, no evidence of bacterial airborne transmission was observed during paper towel dispensing. Conclusion: This pilot study demonstrated that a large community of culturable bacteria, including toxin producers, can be isolated from unused paper towels and that they may be transferred to individuals after handwashing. This may have implications in some industrial and clinical settings as well as in immunocompromised individuals.Publication Restreint Comparison of polycarbonate and polytetrafluoroethylene filters for sampling of airborne bacteriophages(Elsevier, 2010-01-22) Duchaine, Caroline; Gendron, Louis; Moineau, Sylvain; Massé, Daniel; Verreault, Daniel; Rousseau, Geneviève M.Aerosolized coliphage phiX174 and lactococcal phage P008 were sampled with two types of filter, polycarbonate (PC) and polytetrafluoroethylene (PTFE). The recovery was determined by plaque assays and quantitative polymerase chain reaction (qPCR). Recovery by qPCR was greater than by culture and PC filters outperformed PTFE filters both by culture and by qPCR relative recovery. The results of the plaque assays showed that the infectivity of the recovered phages was affected by the aerosolization/air sampling. The presence of viruses in air samples should be determined by culture-independent assays.Publication Accès libre Detection of airborne lactococcal bacteriophages in cheese manufacturing plants(American Society for Microbiology, 2011-01-01) Duchaine, Caroline; Veillette, Marc; Moineau, Sylvain; Massé, Daniel; Verreault, Daniel; Lindsley, William G.; Rousseau, Geneviève M.; Gendron, LouisThe dairy industry adds starter bacterial cultures to heat-treated milk to control the fermentation process during the manufacture of many cheeses. These highly concentrated bacterial populations are susceptible to virulent phages that are ubiquitous in cheese factories. In this study, the dissemination of these phages by the airborne route and their presence on working surfaces were investigated in a cheese factory. Several surfaces were swabbed, and five air samplers (polytetrafluoroethylene filter, polycarbonate filter, BioSampler, Coriolis cyclone sampler, and NIOSH two-stage cyclone bioaerosol personal sampler) were tested. Samples were then analyzed for the presence of two Lactococcus lactis phage groups (936 and c2), and quantification was done by quantitative PCR (qPCR). Both lactococcal phage groups were found on most swabbed surfaces, while airborne phages were detected at concentrations of at least 103 genomes/m3 of air. The NIOSH sampler had the highest rate of air samples with detectable levels of lactococcal phages. This study demonstrates that virulent phages can circulate through the air and that they are ubiquitous in cheese manufacturing facilities.Publication Restreint Airborne porcine circovirus in Canadian swine confinement buildings(Oxford, 2009-09-11) Duchaine, Caroline; Massé, Daniel; Verreault, Daniel; Létourneau, Valérie.; Gagnon, Carl A.; Gendron, LouisPorcine circovirus type 2 has been linked to many diseases, such as postweaning multisystemic wasting syndrome and can be found in most commercial swine confinement buildings around the world. Although the exact role of the virus in the appearance of disease in animals is not fully understood, the mechanisms responsible for the transmission of the virus are currently believed to happen mostly by contact. Nevertheless, the possibility of airborne transmission cannot be rejected. This study investigated the presence of the virus, total bacteria and total dusts in aerosols. Air samples were taken with gelatin filters in swine confinement buildings and were analyzed by quantitative polymerase chain reaction. Interestingly, concentrations of airborne PCV2 of up to 10(7) genomes per cubic meter of air were detected. Airborne dust concentrations were correlated to airborne concentrations of PCV2 and total bacteria. Although the infectivity potential of the airborne viral loads were not evaluated, it is clear that the virus can become airborne in detectable concentrations in commercial swine confinement building environments. The significance of this finding in an epidemiological point of view will need further investigation.Publication Restreint Evaluation of filters for the sampling and quantification of RNA phage aerosols(Elsevier, 2010-08-10) Duchaine, Caroline; Gendron, Louis; Veillette, Marc; Moineau, Sylvain; Verreault, DanielThis study exploits the virulent bacteriophages phi 6 (dsRNA) and MS2 (ssRNA) as surrogates for airborne RNA viruses. Two different filter types, polytetrafluoroethylene (PTFE) and polycarbonate (PC), were tested for their efficiency in collecting aerosolized RNA phages. Two commercial kits were tested for total RNA isolation. Also, heat shock treatments were performed in three different media to obtain the most favorable conditions for reverse transcription assays of dsRNA. Our findings suggest that PC filters are more suitable to recover infectious airborne RNA viruses as determined by plaque assays. Both types of filters were equally efficient in recovering RNA from aerosolized phage phi 6 as established by qRT-PCR. Viral samples should be treated with QIAamp Viral RNA Mini Kit and a 5 min heat shock treatment at 110°C in TE buffer before RT-PCR to maximize detection of phage phi 6. Overall, the infectivity of the recovered phages was severely affected by the aerosolization/air sampling process and the presence of RNA viruses in air samples should be determined by qRT-PCR.