Isolation and purification of Aeromonas sobria cytotonic enterotoxin and beta-haemolysin.
Aeromonas sp., grown in tryptone soya broth supplemented with yeast extract, 0.6%, pH 7.5, and incubated with agitation at 100 oscillations/min for 15 h at 37 degrees C produced optimal amounts of beta-haemolysin and cytotonic enterotoxin. More prolonged incubation resulted in the loss of enterotoxic activity and anion exchange chromatographic analysis indicated the presence of a moiety capable of breaking down the toxin. Anion exchange fast protein liquid chromatography resulted in a single peak of haemolytic activity and two peaks with enterotoxic activity. The cytotonic enterotoxin was purified from the fraction most active in the infant mouse assay; the second peak, which did not cross-react immunologically, may represent a second cytotonic enterotoxin. Neither peak was observed in the chromatographic fractions of filtrates from strains devoid of activity in the infant mouse assay. Purified enterotoxin, estimated to have a mol. wt of 15 kDa by SDS-PAGE, caused fluid accumulation in the infant mouse assay, was non-haemolytic to rabbit erythrocytes, caused an increase in cAMP activity in tissue culture cells and did not cross-react immunologically with components of cholera toxin or the whole toxin. Purified beta-haemolysin had an estimated mol. wt of 55 kDa, lysed rabbit erythrocytes and did not cause fluid accumulation in the infant mouse test.
Gosling
P J
PJ
Public Health Laboratory, Musgrove Park Hospital, Taunton, Somerset.
Turnbull
P C
PC
Lightfoot
N F
NF
Pether
J V
JV
Lewis
R J
RJ
D000333:Aeromonas / Q000378:metabolism / Q000472:pathogenicity*; D000818:Animals; D001427:Bacterial Toxins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003429:Cross Reactions; D000242:Cyclic AMP / Q000032:analysis / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D004768:Enterotoxins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D005677:G(M1) Ganglioside / Q000378:metabolism; D006460:Hemolysin Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D013997:Time Factors; D014774:Virulence / Q000502:physiology
D001427:Bacterial Toxins; D004768:Enterotoxins; D006460:Hemolysin Proteins; D005677:G(M1) Ganglioside; D000242:Cyclic AMP