Colibacillosis is a bacterial disease of great concern in the layer industry causing substantial animal and economic losses worldwide. Breeding for resistance to colibacillosis is an important control strategy that would complement traditional management strategies such as vaccinations and therapeutic treatments. Because antibiotic use in animal production is expected to substantially decrease in the future to meet not only consumer demands but also regulations, it is expected that bacterial diseases such as colibacillosis will become even more important. A challenge test was carried out on 353 pedigreed White Leghorn female chickens of 1 pure line. The birds were inoculated with Escherichia coli at an age of 93 to 95 wk (3 hatches) and symptoms of disease and mortality were constantly monitored for up to 6 d after inoculation. The cumulative mortality was 64%, the majority of which took place in the first 2 d. In the present study, genetic parameters of resistance to colibacillosis were estimated to judge the value of genetic selection. The heritability for survival rate was 0.17+/-0.07. Both for quantitative as well as for qualitative traits of economic interest, there was no remarkable difference between survivors and hens that died. Data showed that it is in principle possible to breed for colibacillosis resistance without penalizing the improvement in other key traits. The B21 marker for MHC in this study was associated with a lower colibacillosis susceptibility. In this study, it was demonstrated that it is possible to breed for Escherichia coli resistance. However, due to the inconveniences of a challenge test, it would be very interesting to make use of MAS.