Leroy C, Delbarre-Ladrat C, Ghillebaert F, Rochet M J, Compère C, Combes D
Laboratoire de Biotechnologie et Molécules Marines, IFREMER Nantes, France.
Lett Appl Microbiol. 2007 Apr;44(4):372-8. doi: 10.1111/j.1472-765X.2006.02103.x.
To develop a method to screen antifouling agents against marine bacterial adhesion as a sensitive, rapid and quantitative microplate fluorescent test.
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-well polystyrene microplate. The 4'6-diamidino-2-phenylindole dye was used to quantify adhered bacteria in each well. The total measured fluorescence in the wells was correlated with the amount of bacteria showing a detection limit of one bacterium per 5 microm(2) and quantifying 2 x 10(7) to 2 x 10(8) bacteria adhered per cm(2). The antifouling properties of three commercial surface-active agents and chlorine were tested by this method in the prevention of adhesion and also in the detachment of already adhered bacteria. The marine bacterial adhesion inhibition rate depending on the agent concentration showed a sigmoid shaped dose-response curve.
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.
In contrast to the 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 the antifouling properties of the products, especially the initial steps of marine biofilm formation.
开发一种筛选抗污剂以抑制海洋细菌黏附的方法，作为一种灵敏、快速且定量的微孔板荧光检测法。
我们的实验方法基于将海洋细菌假交替单胞菌属D41在无菌天然海水中于96孔聚苯乙烯微孔板中单独培养形成的天然生物膜。使用4',6-二脒基-2-苯基吲哚染料对每个孔中黏附的细菌进行定量。孔中测得的总荧光与细菌数量相关，检测限为每5微米²一个细菌，可定量每平方厘米黏附2×10⁷至2×10⁸个细菌。通过该方法测试了三种商用表面活性剂和氯在防止黏附以及去除已黏附细菌方面的抗污性能。取决于试剂浓度的海洋细菌黏附抑制率呈现出S形剂量反应曲线。
该检测方法非常适合在海洋生物膜形成的早期阶段直接在海水中对抗污剂进行快速定量的初步筛选。
与通常检测杀菌活性的抗污产品筛选方法不同，该检测方法更适合筛选具有去除细菌黏附或抑制细菌黏附活性的抗污剂。这种筛选检测关注产品的抗污性能，尤其是海洋生物膜形成的初始阶段。
