Guidelines for the Isolation, Molecular Detection, and Characterization ofBartonellaSpecies

Abstract

Bartonellae are fastidious, facultative, intracellular vector-borne bacteria distributed among mammalian reservoirs worldwide. The pathogenic potential of many Bartonella spp. has increased the interest in these bacteria and advanced their research. Isolation of Bartonella spp. is laborious using classical bacteriological methods and requires specific conditions and prolonged incubation periods. In contrast, molecular methods for detection of Bartonella DNA are considered as more practical and sensitive than the former. Among the molecular methods, the use of real-time PCR assays for primary screening of Bartonella spp., followed by several molecular confirmatory assays, using either conventional or real-time PCR, is recommended. Although primary isolation of Bartonella is a laborious task, we encourage its application to all PCR-positive samples as this is the most reliable proof for the presence of live bacteria. Moreover, a successful trial will enable a broader molecular characterization and speciation of isolated colonies. The present guideline gathers and summarizes recommendations, including advantages and limitations of isolation and molecular detection of Bartonella from mammalian and arthropod samples.

Genetic diversity ofBartonellaspp. in vampire bats from Brazil

“…Among 193 D. rotundus sampled in Peru and Belize, a Bartonella prevalence of 67% was reported (Becker, Bergner, et al, ). Due to their hemotropism, blood is the suitable sample source for Bartonella diagnosis from reservoir animals (Gutiérrez, Vayssier‐Taussat, Buffet, & Harrus, ; Kosoy, Regnery, Kosaya, & Childs, ). When blood is not available, such as in the case of carcasses, the spleen is probably the most preferable internal organ for detection of Bartonella spp.…”

“…When blood is not available, such as in the case of carcasses, the spleen is probably the most preferable internal organ for detection of Bartonella spp. (Gutiérrez et al, ), since bartonellae are retained by the splenic erythrocyte pool and phagocytosed by the splenic macrophages (Deng, Rhun, Lecuelle, Naour, & Vayssier‐Taussat, ). Therefore, the low occurrence of bartonellae found in the present work when compared to those reported in previous studies might be related to the use of liver samples instead of blood or spleen, which were not available in the referred tissue bank.…”

Genotyping ofBartonellabacteria and their animal hosts: current status and perspectives

“…Gutiérrez et al (2017) discuss various methods that are successful in culturing bartonellae and make recommendations for particular sample types (tissue vs ectoparasite). However, one persistent challenge is that many microbes will be challenging to isolate with known culturing techniques.…”

Essential Oils with High Activity against Stationary Phase Bartonella henselae

“…It is laborious using classical bacteriological methods to isolate and culture Bartonella spp. in liquid media especially from clinical samples, which requires specific conditions and prolonged incubation periods [9,10]. Therefore, serology and real-time PCR are often used instead of culture to confirm the diagnosis for rapid Bartonella detection 2 of 17 clinically [9,10].…”

“…in liquid media especially from clinical samples, which requires specific conditions and prolonged incubation periods [9,10]. Therefore, serology and real-time PCR are often used instead of culture to confirm the diagnosis for rapid Bartonella detection 2 of 17 clinically [9,10]. The first-line antibiotics for treating bartonellosis include doxycycline, erythromycin, gentamicin, rifampicin, azithromycin, and ciprofloxacin, as well as some drug combinations like doxycycline plus gentamicin, doxycycline plus rifampin [11,12].…”