Bacteria of the genus Bartonella, a member of the Alphaproteobacteria, are fastidious, gram-negative, aerobic bacilli that comprise numerous species, subspecies, and subtypes. In human and veterinary medicine, species isolation remains a vital component of the diagnostic and therapeutic management of Bartonella infection. We describe a novel, chemically modified, insect-based liquid culture medium that supports the growth of at least seven Bartonella species. This medium will also support cocultures consisting of different Bartonella species, and it facilitated the primary isolation of Bartonella henselae from blood and aqueous fluid of naturally infected cats. This liquid growth medium may provide an advantage over conventional direct blood agar plating for the diagnostic confirmation of bartonellosis.Due to their zoonotic potential, their vector transmission, which includes sandflies, lice, fleas, and ticks, and their frequent adaptation to a mammalian reservoir host, Bartonella species are considered among the newest and most significant emerging pathogens (1,3,8,12,27,29,39,45). These bacteria are highly adapted to a mammalian reservoir host; further, these organisms have been shown to cause a long-lasting intraerythrocytic bacteremia in both humans and animals (11,14,22,24,25,37). Bartonella species are also the causative agents of Carrion's disease (Oroya fever and verruga peruana) (Bartonella bacilliformis) (6), trench fever (B. quintana) (11,40,43), endocarditis (B. elizabethae, B. henselae, B. vinsonii subsp. berkhoffii, B. washoensis, B. clarridgeae) (3, 7, 9, 16, 17, 21, 40, 44), bacillary angiomatosis in immunocompromised patients (B. quintana, B. henselae) (14,30,48), neuroretinitis (B. grahamii) (28), and cat scratch disease (B. henselae, B. clarridgeiae) (5,31,42).Because Bartonella species frequently induce persistent intravascular infections, it has been difficult to attribute chronic disease causation to infection in humans and companion animals; much of this difficulty may be related to the few and often very subtle clinical abnormalities that are reported by a patient or observed in a sick animal. Confirming disease causation is especially difficult in retrospective or prospective animal studies in which Bartonella bacteremia can be detected in overtly healthy, natural reservoir hosts-a paradigm in opposition to Koch's postulates for disease causation (12, 23). Nevertheless, an increasingly diverse spectrum of Bartonella-associated infections have been recognized in people and in dogs due to the development of new approaches to improving serologic and molecular diagnostic testing methods, which prove to be, in most instances, more sensitive than conventional culture methods for the isolation of Bartonella species (13,15,26,30,33,38,41,46,49). Primary isolation of Bartonella species following lysis centrifugation, or freezing of a blood sample, followed by application to a blood agar plate, is the most widely used method for the microbiological diagnosis of bartonellosis. Isolation of Bartonel...