Studies on the impact of interaction of Salmonella enterica serovar Enteritidis and the parasitic nematode Ascaridia galli with the avian host were undertaken with particular emphasis on infection and excretion of these pathogens in two different layer lines. A total of 148 salmonella-free 1-day-old chickens (73 Hellevad and 75 Lohmann Brown) were randomly divided into five groups for each line. Group 1 served as an uninoculated control group. Groups 2 and 3 were infected with A. galli and S. Enteritidis, respectively. Group 4 was first infected with S. Enteritidis and subsequently with A. galli, and vice versa for group 5. The number of chickens excreting S. Enteritidis was significantly higher (P < 0.001) in the groups infected with both S. Enteritidis and A. galli compared with those only infected with S. Enteritidis over time. Furthermore, excretion of S. Enteritidis over time was significantly higher (P < 0.001) in the group first infected with S. Enteritidis and subsequently with A. galli compared with the group infected in the reverse order. No significant differences were observed between the two lines concerning excretion of S. Enteritidis over time in any group (P = 0.61 (group 3), P = 0.73 (group 4), P = 0.31 (group 5)). A. galli established itself significantly better (P = 0.02) in the group first infected with A. galli and subsequently with S. Enteritidis compared with the group infected in the reverse order. Furthermore, the A. galli infection rate was significantly higher (P = 0.02) in Hellevad chickens compared with Lohmann Brown chickens at the end of the experiment.