The indirect fluorescent antibody test was performed to demonstrate the antibody production in the rabbits immunized with free-living amoebas; Acanthamoeba culbertsoni and Naegleria fowleri, and antibody titer changes by immunization duration. Rabbits were immunized with Acanthamoeba culbertsoni and Naegleria fowleri which were cultured axenically in CGVS medium. For experiments, rabbits were divided into two groups; small dose group received 10(4) intravenously with live or dead free-living amoebas trophozoites as an immunizing dose three times with one week interval, and large dose group received 10(6) live or dead trophozoites respectively. The control group received physiologic saline or medium for culture of free-living amoebas intravenously. Antiserum was collected 4 times at interval of 3 days in the first 10 days, and also up to 2 months later. In the group immunized with live Acanthamoeba culbertsoni, fluorescent antibody titer was higher than in the group of dead one, and also in the large dose group than in the small dose group. Antibody titer of anti-Naegleria fowleri serum in the large dose group showed no difference by the source of amoeba antigen; live or dead. But in the small dose group, antibody titer was higher in the immunized with live Naegleria fowleri than in the group with dead one. No cross reactivity was demonstrated between the Acanthamoeba and Naegleria. And no cross reaction was observed when the free-living amoebas antigens were tested against human sera of amoebiasis, paragonimiasis and clonorchiasis.
parasitology-protozoa ; free-living amoeba ; Acanthamoeba culbertsoni ; Naegleria fowleri ; immunology ; fluorescence ; rabbit ; amoebiasis ; paragonimiasis ; clonorchiasis

Rabbits were immunized with free-living amoebas by intravenous injections. The amoebas were Acanthamoeba culbertsoni and Naegleria fowleri and obtained by axenic cultivation in CGVS medium. Each rabbit received 10(6) of Acanthamoeba culbertsoni and 10(5) of Naegleria fowleri trophozoites respectively every other day in three doses and finally one booster dose at 1 week later. Antiserum was collected from thc following day of the booster injection up to 2 months period, and stored at -30 degree C until use. The immobilization test was performed. One drop of amoeba suspension was mixed with the test serum on slide and observed the mobile state under microscope. Maximal immobilizing phenomenon observed in 30 minutes and, then gradually recovered to normal state. Inactivation of antiserum at 56 degree C for 30 minutes did not affect the immobilization phenomenon. The immobilization rates decreased by the serial dilution of antiserum. At dilution more than 1:8, the immobilization was almost the same as in the normal serum. The immobilizing antibody in anti-Acanthamoeba culbertsoni rabbit serum showed highest titre in 3rd day after booster immunization and from first to 6th week in anti-Naegleria fowleri rabbit serum. Cross matching of Acanthamoeba culbertsoni and Naegleria fowleri showed antigenic difference of the two species. It is suggested that the immobilization reaction may be of value as a supplementary test in the diagnosis of primary amoebic meningoencephalitis.
parasitology-protozoa ; free-living amoeba ; Naegleria fowleri ; Acanthamoeba culbertsoni ; rabbit ; immunology