A Bartonella vinsonii berkhoffii typing scheme based upon 16S–23S ITS and Pap31 sequences from dog, coyote, gray fox, and human isolates
Introduction
Bacteria of the genus Bartonella (alpha subdivision of the class Proteobacteria [α-proteobacteria]) are fastidious, gram-negative, aerobic bacilli with more than 20 described species or subspecies [1], [2], [3], [4], [5]. Because of their zoonotic potential; transmission by a wide range of arthropod vectors, including sand flies, lice, fleas, and potentially ticks; and their ability to persistently infect mammalian reservoir hosts, bacteria of the genus Bartonella are considered emerging pathogens [6], [7], [8], [9], [10], [11]. There are three Bartonella vinsonii subspecies: Bartonella vinsonii vinsonii (type strain Baker, ATCC VR-152), Bartonella vinsonii berkhoffii (type strain 93-CO1, ATCC 51672) and Bartonella vinsonii arupensis (type strain OK 94-513, ATCC 700727) [12]. Bartonella vinsonii vinsonii was isolated from a vole in Canada by Baker in 1946 [13], B. vinsonii berkhoffii was isolated from a dog with endocarditis in our laboratory in 1993 [14], [15] and B. vinsonii arupensis was isolated from Peromyscus leucopus from Minnesota and Wisconsin in 1998 and from a cattle rancher from Wyoming in 1994 [12], [16].
Bartonella vinsonii berkhoffii, which was originally isolated from the blood of a dog with intermittent epistaxis and endocarditis, has emerged as an important canine pathogen. The organism has been associated with a wide variety of clinical manifestations including polyarthritis, cutaneous vasculitis, endocarditis, myocarditis, epistaxis, and granulomatous inflammatory disease [8], [14], [15], [17], [18], [19], [20], [21]. In 1998, Kordick et al. described persistent B. vinsonii berkhoffii infection in a sub-clinically infected dog during a 14-month testing period [20]. When this isolate (designated A [20] and Winnie in this manuscript) and two other B. vinsonii berkhoffii isolates obtained from subclinically infected dogs (designated B and C [20]) were subjected to restriction fragment length polymorphism (PCR-RFLP) using HaeIII, the restriction pattern differed from the 93-CO1 isolate obtained from the dog with endocarditis. Since, the isolation of the original B. vinsonii berkhoffii ATCC type strain, we have identified four distinct B. vinsonii berkhoffii types by amplification and sequencing of cultured organisms or PCR amplicons derived from blood or tissue samples obtained from healthy or sick dogs (this study, [15], [19], [20]), from coyotes (Canis latrans) (this study, [10]), from gray foxes (Urocyon cinereoargenteus, this study), from a dog with mitral and aortic valve endocarditis that had not traveled outside of Saskatchewan, Canada (this study) and from human blood samples (this study).
Based upon sequence differences within the 16S–23S ITS region and the bacteriophage associated heme-binding protein Pap31 gene, we propose a genotyping scheme that divides B. vinsonii subsp. berkhoffii isolates into four distinct types. Strain typing should help to better define the reservoir potential, carriership patterns, modes of transmission, and geographic distribution for each B. vinsonii berkhoffii type. In addition, we propose, that B. vinsonii berkhoffii strain 93-CO1 (ATCC 51672), isolated in 1993 at North Carolina State University College of Veterinary Medicine be retrospectively designated as B. vinsonii berkhoffii type I and that strain G7464 isolated from a healthy Greyhound blood donor at the University of Georgia College of Veterinary Medicine (dog B in Ref. [20]), based only on PCR-RFLP of the 16S–23S ITS region) and the isolate designated Winnie in this manuscript (isolate A in Ref. [20]) be retrospectively designated as a type II strains. Type III and type IV stains will be described for the first time as a component of this study.
Section snippets
Strains sources
The sources of B. vinsonii berkhoffii isolates or DNA are summarized in Table 1. A total of 27 strains, including six dog isolates, three gray fox isolates, nine coyote isolates, DNA extracted from six infected aortic valve samples obtained from dogs with endocarditis, and DNA extracted from three human blood samples were sequenced and aligned (Table 1). In addition, the sequence from a previously described human with B. vinsonii berkhoffii endocarditis, deposited in GenBank as accession number
Typing scheme, ITS amplification and sequence alignment
After alignment of 28 sequences from isolates and tissue samples, four clusters could be easily differentiated (Fig. 1, Fig. 2). The proposed genotyping scheme, strains and sources used in this study and the GenBank sequence submission access numbers are summarized in Table 1 for the ITS and Pap31 gene sequences. The 16S–23S ITS region sequences for B. vinsonii berkhoffii type I, type II, type III and type IV, some of the isolates reported in this study, were deposited in the GenBank database
Discussion and conclusions
Bartonella vinsonii berkhoffii sequences derived from the 16S to 23S sRNA intergenic spacer region and from the Pap31 gene amplified from coyote, dog and gray fox and human strains identified differences that allow for a clear division of this subspecies into four distinct strain types. The differences in the16S–23S ITS region are especially important, because there are two specific insertion regions that are variably expressed among the four types. The 37 bp insertion located at position 154 is
Acknowledgements
We thank Dr Susan Taylor from the Western College of Veterinary Medicine, University of Saskatchewan, for providing heart valve tissue from the dog with Bartonella vinsonii berkhoffii type IV infection. We also wish to thank Tonya Lee for editorial assistance and Rick Kasten for technical assistance.
References (28)
- et al.
Spatial analysis of Yersinia pestis and Bartonella vinsonii subsp. berkhoffii seroprevalence in California coyotes (Canis latrans)
Prev Vet Med
(2003) - et al.
Strategy to detect and identify Bartonella species in routine laboratory yields Bartonella henselae from human immunodeficiency virus-positive patient and unique Bartonella strain from his cat
J Clin Microbiol
(1995) - et al.
16S/23S rRNA intergenic spacer regions for phylogenetic analysis, identification, and subtyping of Bartonella species
J Clin Microbiol
(2001) - et al.
Rapid identification and differentiation of Bartonella species using a single-step PCR assay
J Clin Microbiol
(2000) - et al.
Identification of Bartonella (Rochalimaea) species among fastidious gram negative bacteria on the basis of the partial sequence of the citrate-synthase gene
J Clin Microbiol
(1995) - et al.
Current knowledge of Bartonella species
Eur J Clin Microbiol Infect Dis
(1997) - et al.
Bartonella spp. as emerging human pathogens
Clin Microbiol Rev
(1997) - et al.
Survey of Bartonella species infecting intradomicillary animals in the huayllacallan valley, ancash, peru, a region endemic for human bartonellosis
Am J Trop Med Hyg
(1999) - et al.
Bartonella infection in animals: carriership, reservoir potential, pathogenicity, and zoonotic potential for human infection
Clin Microbiol Rev
(2000) - et al.
Bartonella spp. isolated from wild and domestic ruminants in North America
Emerg Infect Dis
(2000)
Coyotes (Canis latrans) as the reservoir for a human pathogenic Bartonella sp.: molecular epidemiology of Bartonella vinsonii subsp. berkhoffii infection in coyotes from central coastal California
J Clin Microbiol
Bartonella (Rochalimaea) infections: beyond cat scratch
Annu Rev Med
Isolation of a new subspecies, Bartonella vinsonii subsp. arupensis, from a cattle rancher: identity with isolates found in conjunction with Borrelia burgdorferi and Babesia microti among naturally infected mice
J Clin Microbiol
A rickettsial infection in Canadian voles
J Exp Med
Cited by (61)
Polymerase chain reaction detection of Bartonella spp. in dogs from Spain with blood culture-negative infectious endocarditis
2018, Journal of Veterinary CardiologyCitation Excerpt :The detected species comprised of B. vinsonii subsp. berkhoffii, B. rochalimae, and B. koehlerae, which are similar to the canine Bartonella spp. previously described in endocarditis cases [3,7–26,33]. Unexpectedly, B. rochalimae (6 out of 8) was found as one of the main causes of PCR positive dogs with blood culture-negative endocarditis in the present study, rather than B. vinsonii subsp. berkhoffii which was the species more frequently detected in previous publications [3,7–26,33]. This higher prevalence of B. rochalimae could reflect differences in the type of samples used, Bartonella spp. epidemiology of different areas, and exposure to different risk factors among the study populations.
Bartonella Species and Vascular Pathology
2016, Vascular Responses to PathogensBartonellosis: Zoonotic overview
2014, Revue Francophone des LaboratoiresFlea species infesting dogs in Florida and Bartonella spp. prevalence rates
2014, Veterinary ParasitologyCitation Excerpt :Bacterial species and strain were defined by comparing similarities with reference sequences chosen from the GenBank database using Basic Local Alignment Search Tool. DNA from samples for which sequencing revealed homology to Bvb was submitted to North Carolina State University for genotyping as described previously (Kordick et al., 1996; Maggi et al., 2006). The flea genera were determined for 43 dogs; 28 dogs (65.1%) were infested with C. felis alone, 8 dogs (18.6%) were infested with Pulex spp. alone, and 7 dogs (16.3%) were infested by both C. felis and Pulex spp.
Experimental infection of dogs with various Bartonella species or subspecies isolated from their natural reservoir
2014, Veterinary MicrobiologyCitation Excerpt :At least two different genotypes of B. henselae have been identified (Boulouis et al., 2005) and it has been suggested that genotype I may infect humans more frequently than genotype II. Similarly, at least 4 genotypes of B. v. berkhoffii have been described (Maggi et al., 2006). We report on experimental infections of research dogs with B. henselae, B. v. berkhoffii and B. rochalimae strains isolated from naturally infected domestic or wild reservoirs and compare these data with three previous experimental infections with B. v. berkhoffii (Balakrishnan et al, 2013; Pappalardo et al., 2001) and B. rochalimae (Chomel et al., 2009).
Bartonella bacteria in nature: Where does population variability end and a species start?
2012, Infection, Genetics and Evolution