A capillary tube feeding (CTF) system was adapted for studying the interaction between Dermacentor variabilis (Say) and the rickettsial cattle pathogen Anaplasma marginale Theiler. A. marginale undergoes a complex developmental cycle in ticks that begins in midguts and ends by transmission from salivary glands. In this CTF system, male D. variabilis were fed A. marginale-infected blood or cultured tick cells. Ticks that fed on highly rickettsemic calves developed midgut and salivary gland infections as detected by PCR, whereas ticks that were fed from capillary tubes on the same blood developed only midgut infections. An unexpected result of capillary tube feeding was that antibodies against the A. marginale adhesin, major surface protein la, enhanced midgut infections and caused cell culture-derived A. marginale to infect midguts. Another unexpected result was the infection of the midguts of the nonvector tick Amblyomma americanum (L.), after capillary tube feeding on infected bovine blood. The gut cell response of ticks to A. marginale, as determined from SDS-polyacrylamide gel electrophoresis protein profiles, did not differ when ticks were fed infected or uninfected cells from capillary tubes. Selected protein bands, as identified by tryptic digestion-mass spectrometry, contained mostly proteins of bovine origin, including bovine albumin, undigested alpha- and beta-chain hemoglobin and hemoglobin fragments. Although infection of ticks by A. marginale CTF system was not the same as infection by feeding on cattle, the results obtained demonstrated the potential use of this system for identifying aspects of pathogen-vector interactions that are not readily recognized in naturally feeding ticks.