The relationship between preinfection functions of blood neutrophils and outcome of experimental Escherichia coli mastitis was studied in 11 cows. Random migration, chemotaxis, phagocytosis, and chemiluminescence by neutrophils were determined in white blood cell suspensions, and in purified neutrophil suspensions. The course of E. coli mastitis (10(4) E. coli 0:157 in rear quarters) was monitored using clinical parameters, counts of E. coli in mastitic secretion, and milk production. Regressions were calculated for areas under curves of these parameters and preinfection activities of neutrophils. Chemiluminescence by nonstimulated neutrophils in white blood cell suspensions was negatively correlated with counts of E. coli in secretion and with losses in milk production. The chemotactic differential in white blood cell suspensions minus the chemotactic differential in purified suspensions of neutrophils referred to as delta varied from -.66 to +.50, indicating, respectively, inhibition and stimulation of chemotactic activity of neutrophils in white blood cell suspensions. Delta correlated negatively with counts of E. coli in mastitis secretion, inhibition of the amplitude of rumen contractions, and losses in milk production. We hypothesize that a factor in white blood cell suspensions may be involved in the down-regulation of the migratory response of neutrophils during E. coli mastitis.