The flagella staining technique to be described is a modification and perfection of that originally published by the author in 1930. The arrangement and, to a lesser extent, the shape of flagella are of considerable taxonomic importance. Flagellar arrangement seems to be a characteristic of sufficient stability and distinctiveness to justify the classification of flagellated bacteria into several groups. Only rarely, and then mainly with very poorly flagellated bacteria, is the flagellar arrangement difficult to determine. Mutation from one type of flagellar arrangement to another has not, to my knowledge, been described and confirmed. Variation from flagellated to nonflagellated types is common in some groups of bacteria, and several instances have been recorded of loss of motility without loss of flagella (Knaysi, 1933; Kauffmann, 1939; Edwards et al., 1946; Hirsch, 1947; Friewer and Leifson, 1951). The shape of the flagella on an organism, while usually quite characteristic, may be modified somewhat by the environment and also may undergo distinct mutational variation (personal observation). A comprehensive review of the literature is given by Houwink and Van Iterson (1950).

Staining, shape and arrangement of bacterial flagella.

Motility on surfaces is an important mechanism for bacterial colonization of new environments. In this report, we describe detection of rapid surface motility in the wild-type Bacillus subtilis Marburg strain, but not in several B. subtilis 168 derivatives. Motility involved formation of rapidly spreading dendritic structures, followed by profuse surface colonies if sufficient potassium ion was present. Potassium ion stimulated surfactin secretion, and the role of surfactin in surface motility was confirmed by deletion of a surfactin synthase gene. Significantly, this motility was independent of flagella. These results demonstrate that wild-type B. subtilis strains can use both swimming and sliding-type mechanisms to move across surfaces.

Rapid Surface Motility in Bacillus subtilis Is Dependent on Extracellular Surfactin and Potassium Ion

Laboratory aquarium experiments demonstrated that Vibrio strain AK-1 caused rapid and extensive bleaching of the coral Oculina patagonica at 29°C, slower and less-complete bleaching at 23°C, and no bleaching at 16°C. At 29°C, the application of approximately 100 Vibrio strain AK-1 cells directly onto the coral caused 50 and 83% bleaching after 10 and 20 days, respectively. At 16°C, there was no bleaching, even with an initial inoculum of 1.2 × 108 bacteria. To begin to understand the effect of seawater temperature on bleaching of O. patagonica by Vibrio strain AK-1, adhesion of the bacteria to the coral as a function of temperature was studied. Inoculation of 107 Vibrio strain AK-1 organisms into flasks containing 20 ml of seawater at 25°C and a fragment of O. patagonica resulted in net levels of bacterial adhesion to the coral of 45, 78, and 84% after 2, 6, and 8 h, respectively. The adhesion was inhibited 65% by 0.001% d-galactose and 94% by 0.001% methyl-β-d-galactopyranoside (β-M-Gal). After the incubation of Vibrio strain AK-1 with the coral for 6 h, 42% of the input bacteria were released from the coral with 0.01% β-M-Gal, compared to less than 0.2% when β-M-Gal was present during the adhesion step. Adhesion did not occur when Vibrio strain AK-1 was grown at 16°C, regardless of whether the corals were maintained at 16 or 25°C, whereas bacteria grown at 25°C adhered to corals maintained at 16 or 25°C. Bacteria grown at 25°C adhered avidly to Sepharose beads containing covalently bound β-d-galactopyranoside but failed to bind if grown at 16°C. These data suggest that elevated seawater temperatures may cause coral bleaching by allowing for the expression of adhesin genes of Vibrio strain AK-1.

https://aem.asm.org/content/aem/64/4/1379.full.pdf

Effect of Temperature on Adhesion of Vibrio Strain AK-1 to Oculina patagonica and on Coral Bleaching.

In this study, we have reevaluated the taxonomic position of biotype 2 of Vibrio vulnificus. For this purpose, we have biochemically and serologically characterized 83 biotype 2 strains from diseased eels, comparing them with 17 biotype 1 strains from different sources. Selected strains were also molecularly analyzed and tested for eel and mouse pathogenicity. Results have shown that biotype 2 (i) is biochemically homogeneous, indole production being the main trait that distinguishes it from biotype 1, (ii) presents small variations in DNA restriction profiles and outer membrane protein patterns, some proteins being immunologically related to outer membrane proteins from biotype 1, (iii) expresses a common lipopolysaccharide (LPS) profile, which is immunologically identical among strains and distinct from that of LPS of tested biotype 1 strains, and (iv) contains at least two high-Mr plasmids. Regarding host range, we have confirmed that both biotypes are pathogenic for mice but only biotype 2 is pathogenic for eels. On the basis of these data, we propose that biotype 2 of V. vulnificus constitutes an LPS-based O serogroup which is phenotypically homogeneous and pathogenic for eels. In this article, the serogroup is designated serogroup E (for eels).

Phenotypic characterization of Vibrio vulnificus biotype 2, a lipopolysaccharide-based homogeneous O serogroup within Vibrio vulnificus

The kinetics of adhesion of Vibrio strains isolated from diseased fish to skin mucus of gilt-head sea bream was studied. A modified Langmuir adsorption isotherm was calculated, and the results obtained indicate that the strains tested (Vibrio alginolyticus DP1HE4 and Vibrio anguillarum-like DC12R8 and DC12R9) showed a saturation kinetics except for V. alginolyticus (CAN), which showed a proportional adsorption kinetics. The adhesive capability for skin mucus does not seem to be an essential virulence factor of pathogenic strains of Vibrio, since this specific interaction depended on several environmental factors, temperature and salinity being the most important. However, the absence of an inhibitory effect of mucus on the pathogenic microorganisms, and the capability of the Vibrio strains to utilize mucus as a carbon source, could favor their settlement on the skin with a potential for infection of cultured, stressed fish.

https://aem.asm.org/content/aem/62/10/3650.full.pdf

Kinetics of adhesion of selected fish-pathogenic Vibrio strains of skin mucus of gilt-head sea bream (Sparus aurata L.).

Staining bacterial flagella.

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Staining bacterial flagella.