Molecular detection of Bartonella spp. in deer keds Lipoptena cervi and Lipoptena fortisetosa in Lithuania

Abstract

Bartonella spp. are gram-negative facultative intracellular bacteria that are highly adapted to their mammalian reservoir hosts, in which they cause chronic relapsing intraerythrocytic bacteraemia. Several Bartonella spp. caused disease in humans and animals named bartonellosis. In ruminants are commonly found Bartonella schoenbuchensis, Bartonella bovis, Bartonella capreoli and Bartonella chomelii species. Deer keds, blood-sucking haematophagous ectoparasites of cervids, are vectors of Bartonella species associated with ruminants. Given that deer keds accidentally bite humans there is a potential risk for occasional transmission of human pathogenic B. schoenbuchensis. The aim of this study was to investigate the prevalence of Bartonella spp. in two species of deer keds Lipoptena cervi and Lipoptena fortisetosa and genetically characterize the bacteria using PCR and sequence analysis. A total of 292 deer keds were collected from roe deer (n=5), red deer (n=1) and moose (n=2) in Lithuania during 2017. Species of deer keds was identified by morphological examination using identification keys and microscope. DNA from deer keds was isolated using Genomic DNA Purification Kit, PCR, nested PCR assays and sequence analysis based on rpoB gene and 16S-23S rRNA intergenic spacer region (ITS) were used for detection and molecular characterization of Bartonella spp. Obtained Bartonella sequences were analyzed by comparing similarities with other sequences deposited in the GenBank database using BLASTn identity searches and MEGA 6.0 software. The phylogenetic tree was constructed using Neighbor-Joining method and Kimura 2 parameter model. Bartonella DNA was detected in 48 (98%) from 49 pools of L. cervi and 37 (74%) from 50 pools of L. fortisetosa deer keds. Analyzed Bartonella ITS and rpoB sequences derived from L. cervi and L. fortisetosa showed 96-99 % similarity with B.schoenbuchensis, B.chomelii and B.capreoli species. Bartonella sequences were heterogenic: [...]