Laboratory Maintenance of Bartonella quintana

Abstract

Trench fever is the common name for the acute febrile syndrome associated with a Bartonella quintana bacterial infection. The focus of this unit is to describe reliable methods for cultivation and cryopreservation of B. quintana and can be applied to cultivation and preservation of all Bartonella. Detailed recipes for preparation of three types of semisolid media are also included.

Combining Culture Techniques for Bartonella: the Best of Both Worlds

“…B. quintana was the only strain for which the presence of mammalian cells in the culture (M10-V medium) was a significant benefit to growth compared to the growth achieved by culture under the same conditions but with cell-free (M10) medium. These results agree with those from previous work suggesting that under mammalian cell culture-based conditions, cell coculture is important for B. quintana growth (3,22). Recent developments, however, have shown that cell culture is unnecessary when insect cell culture-based medium is used (28,32).…”

“…The data were analyzed for each strain and between conditions at the time course endpoint of 9 days or at the peak of growth using the Kruskal-Wallis or Wilcoxon rank sum tests with 2 approximations. The day 9 endpoint was used for the purpose of comparing media and cell presence, as this was most biologically relevant to Bartonella culturing protocols, which usually require incubation for from 7 to 28 days (3,22,24). The peak growth of each strain was compared between temperatures at day 9, as it was the most logical point to show temperature-dependent differences because of such a discrepancy in growth at day 9, when the growth either plateaued or dramatically declined.…”

“…Numerous Bartonella species have been associated with clinical illnesses ranging from mild skin lesions to more severe manifestations, including persistent fevers, neurological symptoms, and endocarditis (6, 10). These bacteria can be fastidious under current laboratory conditions, and attempts to isolate cells from pure culture of biological specimens are often unsuccessful, despite positive molecular detection (1, 7).The majority of culturing methods for Bartonella species found in the literature focus on growth requirements for the two species commonly associated with human infection, Bartonella henselae and B. quintana (3,9,22,27,35). Through a review of the literature on methods for Bartonella isolation from biological samples, we found many differences in the culturing protocols.…”

“…The majority of culturing methods for Bartonella species found in the literature focus on growth requirements for the two species commonly associated with human infection, Bartonella henselae and B. quintana (3,9,22,27,35). Through a review of the literature on methods for Bartonella isolation from biological samples, we found many differences in the culturing protocols.…”

Colonization of Lutzomyia verrucarum and Lutzomyia longipalpis Sand Flies (Diptera: Psychodidae) by Bartonella bacilliformis, the Etiologic Agent of Carrión’s Disease

“…Bartonella bacilliformis was cultivated as previously described for B . quintana except the incubation temperature was 30°C and 5% CO 2 was omitted [ 13 ]. Briefly, h eart i nfusion b roth + sheep b lood (HIB-B) plates consisted of heart infusion broth containing 4% sheep blood, 2% sheep serum and 1.5% agar.…”