Challenge testing of ready-to-eat (RTE) foods with Listeria monocytogenes is recommended to assess the potential for growth. The present study was undertaken to evaluate a protocol for challenge testing applied to RTE cooked meat products. In order to choose L. monocytogenes strains with a representative behaviour, initially, the variability of the response of multiple L. monocytogenes strains of human and food origin to different stress and growth conditions was established. The strains were not inhibited in their growth at moderate acid pH (5.25) and the four strains tested in particular showed a similar acid-adaptive response. Growth of the various strains under four different combined stress conditions indicated that no L. monocytogenes strain had consistently significant longer or shorter lag phase or higher or lower maximum specific growth rates. The effect of choice of strain and history (pre-incubation temperature 7 or 30 degrees C) on growth of L. monocytogenes under optimum conditions (Brain Heart Infusion, BHI) and modified BHI simulating conditions of cooked ham and paté was studied. In general, all four L. monocytogenes strains behaved similarly. In BHI, no difference in lag phase was observed for the cold-adapted and standard inoculum, whereas in BHI adjusted to ham and pâté conditions, a ca. 40-h reduction of the lag phase was noted for the cold-adapted inoculum. Subsequently, microbial challenge testing of L. monocytogenes in modified atmosphere packaged sliced cooked ham and paté was performed. A mixed inoculum of four L. monocytogenes strains and an inoculum level of ca. 1-10 cfu/g was used. On vacuum packed sliced cooked ham, the concentration of 100 cfu/g, the safety limit considered as low risk for causing listeriosis, was exceeded after 5 days whereas ca. 10(5) cfu/g were obtained after 14 days when also LAB spoilers reached unacceptable numbers (ca. 10(7) cfu/g) whether standard or cold-adapted inoculum was used. The concentration of sodium lactate determined the opportunities for growth of L. monocytogenes in pâté. If growth of L. monocytogenes in pâté was noticed, the threshold of 100 cfu/ml was crossed earlier for the cold-adapted inoculum compared to the standard inoculum.