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A marine bacterial adhesion microplate test using the DAPI fluorescent dye: a new method to screen antifouling agents ArchiMer
Leroy, Celine; Delbarre Ladrat, Christine; Ghillebaert, F; Rochet, Marie-joelle; Compere, Chantal; Combes, D.
Aims: To develop a method to screen antifouling agents against marine bacterial adhesion as a sensitive, rapid, quantitative microplate fluorescent test. Methods and Results: Our experimental method is based on a natural biofilm formed by mono-incubation of the marine bacterium Pseudoalteromonas sp. D41 in sterile natural sea water in a 96 wells polystyrene microplate. The 4'6-diamidino-2-phenylindole (DAPI) dye was used to quantify adhered bacteria in each well. Total measured fluorescence in wells was correlated with the amount of bacteria showing a detection limit of one bacterium per 5 µm2 and quantifying 2 x 107 to 2 x 108 bacteria adhered per cm2. The antifouling properties of three commercial surface active agents and chlorine were tested by this...
Tipo: Text Palavras-chave: Antifouling; Pseudoalteromonas; Screening; Microtiter plate; Adhesion; DAPI; Marine biofilm.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2006/publication-2017.pdf
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Effects of commercial enzymes on the adhesion of a marine biofilm-forming bacterium ArchiMer
Leroy, C.; Delbarre Ladrat, Christine; Ghillebaert, F.; Compere, Chantal; Combes, D..
The antifouling potential of commercial hydrolases, four proteases, seven glycosidases and one lipase was evaluated on the adhesion of marine Pseudoalteromonas sp. D41. The experimental method, adapted to screen antifouling agents, was based on bacterial adhesion in natural sterile sea water in a microtiter plate and on total biomass quantification by the fluorescent dye DAPI (4[prime]6-diamidino-2-phenylindole). Savinase (subtilisin) was the most effective hydrolase in both the prevention of bacterial adhesion and the removal of adhered bacteria. However, some enzymatic preparations tested such as Amano protease were not only ineffective but also increased the number of adhered bacterial cells. Enumeration using epifluorescence microscopy of CTC...
Tipo: Text Palavras-chave: Marine biofilm; Antifouling; Pseudoalteromonas; Enzymes; Adhesion.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/publication-6140.pdf
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Lutte contre les salissures marines : approche par procédés enzymatiques. ArchiMer
Leroy, Celine.
Fouling on marine underwater surfaces causes critical and economic problems such as important material biodamages and industrial performances reduction. We chose to test antifouling potential of enzymatic commercial preparations like hydrolases (proteases, glycosidases and lipases) in order to inhibit the first fouling adhesion step: bacterial biofilm formation. An evaluation test of antifouling properties onto marine bacterial adhesion was designed using a mono-incubation of Pseudoalteromonas sp. D41 in microtiter plate and in sterile natural sea water. This test was adapted to screen agents for bacterial adhesion removal or inhibition activities and allowed to test enzymatic preparations toxicity on non adhered bacteria. Inhibition rates according to...
Tipo: Text Palavras-chave: Fouling; Adhesion; Antifouling; Hydrolases; EPS; DAPI; Microtiter plate; Screening; Pseudoalteromonas sp.; Marine biofilm; Salissure marine; Adhésion; Antisalissure; Hydrolases; EPS; DAPI; Microplaque; Criblage; Pseudoalteromonas sp.; Biofilm marin.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/these-1099.pdf
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