Complete Genome Sequence of Bartonella ancashensis Strain 20.00, Isolated from the Blood of a Patient with Verruga Peruana.

Here we present the complete genome sequence of Bartonella ancashensis strain 20.00, isolated from the blood of a Peruvian patient with verruga peruana, known as Carrion's disease. Bartonella ancashensis is a Gram-negative bacillus, phylogenetically most similar to Bartonella bacilliformis, the causative agent of Oroya fever and verruga peruana.

GENOME ANNOUNCEMENT

Members of the genus Bartonella are Gram-negative bacilli that are found worldwide and are associated with animal and human diseases (1). The most common diseases associated with Bartonella species are trench fever, cat scratch disease, and the biphasic illness Carrion’s disease, which are caused by B. quintana, B. henselae, and B. bacilliformis, respectively (2–4). More recently, B. henselae and B. quintana have been associated with bacillary angiomatosis, and more than eight Bartonella species have been associated with either febrile illnesses or endocarditis (2, 4, 5).

In a 2003 study involving 127 patients with verruga peruana in the Peruvian Ancash mountain area, two patients were found to be infected with a Bartonella species distinct from any known Bartonella species. This determination was based on the sequences of the gltA, rpoB, ftsZ, groEL, and rrs genes and the 16s-23s intergenic internal transcribed spacer (ITS) region. The genetic differences and culture characteristics of the strain meet the taxonomic criteria to classify this non-B. bacilliformis pathogen as a new species, designated Bartonella ancashensis. The nomenclature has been accepted by the International Journal of Systematic and Evolutionary Microbiology, and B. ancashensis strain 20.00 was accepted by both ATCC and DSMZ for archiving and distribution for research use (6–8).

The genome of B. ancashensis strain 20.00 was sequenced using Roche 454 next-generation sequencing technology (Roche 454 Life Sciences, Branford, CT). A total of 198,990 filtered reads consisting of 57.4 Mb of data were assembled into 94 contigs with average sequence coverage of 41.7× using GS Assembler software (Newbler) version 2.5.3. Final assembly was verified using a whole-genome AflII restriction map generated with the Argus system (OpGen, Gaithersburg, MD) (9). B. ancashensis strain 20.00 has a circular genome of 1,466,048 bp and has a G+C content of 38.4%, which is similar to the genome of the Bartonella prototype strain, B. bacilliformis KC583 (NC_008783.1), which is 1,445,021 nucleotides in length with a G+C content of 38.2%.

Whole-genome annotation was performed using RAST (Rapid Annotation using Subsystem Technology) (http://www.nmpdr.org/FIG/wiki/view.cgi/FIG/RapidAnnotationServer); structural and functional annotation was completed using the IGS Annotation Engine (http://ae.igs.umaryland.edu/cgi/index.cgi). Genome annotations were reviewed and finalized using Genome Viewer (http://www.nmpdr.org/FIG/wiki/view.cgi/FIG/GenomeViewer) and Manatee (http://manatee.sourceforge.net/) (10, 11). The B. ancashensis strain 20.00 genome contains 1,346 putative protein-encoding genes, of which 79.1% are found to have homologs in B. bacilliformis. Like B. bacilliformis, B. ancashensis strain 20.00 encodes flagellar proteins. However, unlike B. bacilliformis, B. ancashensis strain 20.00 encodes VirB/D4 type IV secretion system proteins most similar to those of B. clarridgeiae and B. rochalimae, both of which have been implicated in human disease. Further, B. ancashensis possesses a family of virulence-modulating proteins which are present only in B. australis and human-pathogenic Leptospira species (12–16).

Here we have reported the first complete genome sequence for the new pathogen B. ancashensis. Further B. ancashensis genome studies and comparisons will elucidate factors involved in virulence and pathogenicity of not only B. ancashensis, but also the Bartonella genus as a whole.

Nucleotide sequence accession number.

This whole-genome shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession number CP010401.