Probing for enterotoxigenicity among the salmonellae: an evaluation of biological assays

Sixty-eight Salmonella strains representing 39 serotypes were variously screened for enterotoxigenicity by using the Chinese hamster ovary (CHO), Y1 adrenal, and Vero cell tests, rabbit skin tests for delayed permeabiltity factor (DPF) and rapid permeability factor (RPF), the rabbit ileal loop test, and the infant mouse test. An iron-sufficient medium, YT-1, and a deferrated medium, DF, were compared. Of the culture supernatant fluids of strains grown in DF medium, 66% yielded positive reactions in the CHO cell test compared with only 10% with TY-1 medium. The corresponding performances with supernatant fluids of DF medium cultures in Y1 adrenal and Vero cell tests were 85 and 69% positive, respectively. The overall agreement between the Y1 adrenal or CHO cell test and the rabbit skin test for DPF, i.e., positive or negative in both tests, was about 70%. Positivity in DPF tests was a better predictor of positivity in either the Y1 adrenal or rabbit ileal loop test than vice versa. CHO cell, DPF, and rabbit ileal loop reactivities of unheated culture filtrates were each neutralized by anticholera antitoxin. Only four strains gave positive reactions in the infant mouse test, whereas up to 66% were positive for RPF in rabbit skin, based on positivity in Ty-1 or DR medium or both. DPF and RPF were produced by 35% of the strains. Of the 28 isolates from human stools, 82 and 92% and all of 11 strains tested were positive in the DPF, Y1 adrenal cell, and rabbit ileal loop tests, respectively. The corresponding data for 17 sewage isolates, representing 17 different serotypes rarely isolated from human stools in Sweden, were 63 and 69% and 8 of 8 tested. On the basis of this investigation, rabbit skin tests for both DPF and RPF provide the most reliable means of screening for enterotoxigenicity among salmonellae.