A marine bacterial adhesion microplate test using the DAPI fluorescent dye: a new method to screen antifouling agents

Type Article
Date 2007-04
Language English
Author(s) Leroy Celine1, 4, Delbarre Ladrat ChristineORCID1, Ghillebaert F4, Rochet Marie-Joelle1, Compere ChantalORCID2, Combes D3
Affiliation(s) 1 : IFREMER, Lab Biotechnol & Mol Marines, Nantes, France.
2 : IFREMER, Serv Interfaces & Capteurs, Brest, France.
3 : INSA, Lab Biotechnol Bioprocedes, Toulouse, France.
Source Letters in Applied Microbiology (0266-8254) (Blackwell science), 2007-04 , Vol. 44 , N. 4 , P. 372-378
DOI 10.1111/j.1472-765X.2006.02103.x
WOS© Times Cited 35
Keyword(s) Antifouling, Pseudoalteromonas, Screening, Microtiter plate, Adhesion, DAPI, Marine biofilm
Abstract 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 method in prevention of adhesion and also in detachment of already adhered bacteria. The marine bacterial adhesion inhibition rate depending on agent concentration showed a sigmoid shape like dose-response curve.

Conclusions: This test is well adapted for a rapid and quantitative first screening of antifouling agents directly in seawater in the early steps of marine biofilm formation.

Significance and impact of the study: In contrast with usual screenings of antifouling products which detect a bactericidal activity, this test is more appropriate to screen antifouling agents for bacterial adhesion removal or bacterial adhesion inhibition activities. This screening test focuses on antifouling properties of products, especially the first steps of marine biofilm formation.
Full Text
File Pages Size Access
publication-2017.pdf 8 93 KB Open access
Top of the page