Identification of capsule, biofilm, lateral flagellum, and type IV pili in Vibrio mimicus strains.

Vibrio mimicus is a bacterium that causes gastroenteritis; it is closely related to Vibrio cholerae, and can cause acute diarrhea like cholera- or dysentery-type diarrhea. It is distributed worldwide. Factors associated with virulence (such as hemolysins, enterotoxins, proteases, phospholipases, aerobactin, and hemagglutinin) have been identified; however, its pathogenicity mechanism is still unknown. In pathogenic Vibrio species such as V. cholerae, Vibrio. parahaemolyticus and Vibrio vulnificus, capsule, biofilms, lateral flagellum, and type IV pili are structures described as essential for pathogenicity. These structures had not been described in V. mimicus until this work. We used 20 V. mimicus strains isolated from water (6), oyster (9), and fish (5) samples and we were able to identify the capsule, biofilm, lateral flagellum, and type IV pili through phenotypic tests, electron microscopy, PCR, and sequencing. In all tested strains, we observed and identified the presence of capsular exopolysaccharide, biofilm formation in an in vitro model, as well as swarming, multiple flagellation, and pili. In addition, we identified homologous genes to those described in other bacteria of the genus in which these structures have been found. Identification of these structures in V. mimicus is a contribution to the biology of this organism and can help to reveal its pathogenic behavior.

Microwave irradiation for shortening the processing time of samples of flagellated bacteria for scanning electron microscopy.

Microwave irradiation (MWI) has been applied to the development of rapid methods to process biological samples for scanning electron microscopy (SEM). In this paper we propose two simple and quick techniques for processing bacteria (Proteus mirabilis and Vibrio mimicus) for SEM using MWI. In the simplest methodology, the bacteria were placed on a cover-glass, air-dried, and submitted to conductivity stain. The reagent used for the conductivity stain was the mordant of a light microscopy staining method (10 ml of 5% carbolic acid solution, 2 g of tannic acid, and 10 ml of saturated aluminum sulfate 12-H2O). In the second method the samples were double fixed (glutaraldehyde and then osmium), submitted to conductivity stain, dehydrated through a series of ethanol solutions of increasing concentration, treated with hexamethyldisilazine (HMDS), and dried at 35 degrees C for 5 minutes. In both methods the steps from fixation to treatment with HMDS were done under MWI for 2 minutes in an ice-water bath, in order to dissipate the heat generated by the MWI. Although both techniques preserve bacterial morphology adequately, the latter, technique showed the best preservation, including the appearance of flagella, and that process was completed in less than 2 hours at temperatures of MWI between 4 to 5 degrees C.