Bartonella henselae, the predominant cause of cat scratch disease, and Bartonella quintana, the cause of trench fever, are closely related Bartonella species that induce cross-reactivity when cat or human sera are tested using an indirect immunofluorescence antibody (IFA) test. Cats are the natural reservoir for B. henselae, whereas a mammalian reservoir host for B. quintana has not been identified. Serum samples from 114 cats from Israel and 114 cats from North Carolina were tested by IFA for seroreactivity to B. henselae and B. quintana antigens. Similar numbers of cats from Israel [45 (39.5%)] and from North Carolina [46(40.4%)] were seroreactive to both antigens, however, as compared to cats from North Carolina [8 (7%)], a significantly (P = 0.001) larger number of cats from Israel were seroreactive to B. quintana antigen only [23 (20.2%)]. In addition, mean antibody titers were lower to B. henselae than to B. quintana (P = 0.0001) in the cats from Israel, whereas similar mean titers to both antigens were identified in cats from North Carolina. Absorption of serum using whole B. henselae organisms resulted in a significantly greater (P = 0.0001) decrease in antibody titer to B. henselae between absorbed and non-absorbed sera, as compared to the decrease in antibody titer following absorption with whole B. quintana organisms. There was a similar decrease in antibody titer in sera from cats experimentally infected with B. henselae and in cats naturally exposed to Bartonella species from Israel and North Carolina. Our results indicate that absorption of serum will, in most instances, distinguish species-specific reactivity by IFA to B. henselae from cross-reactivity to B. quintana in cats experimentally infected with B. henselae. The data support the conclusion that B. henselae is the principal Bartonella species responsible for seroreactivity against B. henselae and B. quintana in naturally exposed cats from Israel or North Carolina. It also suggests that in Israel, cats are exposed to one or more antigenically different Bartonella species, sub-species or strains, that seroreact by IFA more intensely with B. quintana antigen.