To determine the most appropriate dose in drinking water of the disinfectant N-alkyl dimethyl benzyl ammonium chloride (TIMSEN), a fowl typhoid challenge trial was carried out using 21-day-old Salmonella-free chickens. In a pretrial, performed with six groups of 10 chickens each, it was shown that the disinfectant was atoxic and safe when administered during 15 days at doses of 0 ppm, 200 ppm, 400 ppm, 800 ppm, 1600 ppm, or 3200 ppm. Thereafter, a challenge trial was performed with 390 chickens divided into six groups of 65 birds each. Chickens were treated during 9 days with oral doses of 0 ppm, 25 ppm, 50 ppm, 100 ppm, 250 ppm, and 500 ppm (groups identified as A, B, C, D, E, and F, respectively). Twenty-four hours after the beginning of the treatment, 30 chickens of each group were orally inoculated with 10(9) colony-forming units of Salmonella Gallinarum strain INTA 91 per bird. The remaining 35 unchallenged chickens from each group were left in the same cage in close contact with the other challenged birds of their group. All surviving chickens were sacrificed 8 days after challenge. Livers from all birds were examined for the presence of Salmonella Gallinarum. Salmonella Gallinarum was isolated from all challenged chickens and from some unchallenged chickens of groups A (4/35), E (3/35), and F (6/35), but not from any unchallenged chicken from groups B, C, and D. Doses of 50 ppm (group C) significantly reduced mortality (30%) in comparison with the untreated control group A (65%). Mortality in group B (45%) was not significantly different from group C. Administration of higher doses of the disinfectant resulted in significantly higher mortality rates, 70% in group E and 85% in groups D and F. Increased infection and mortality rates of groups D, E, and F might have been caused by inhibition of the protective action of the normal gut flora. To reduce horizontal infection and mortality, the manufacturer's prescribed oral dose of 25 ppm may be increased up to 50 ppm. Nevertheless, higher doses should be avoided because they may cause horizontal increased spread of the disease and mortality.