Comparative genomics of first available bovine Anaplasma phagocytophilum genome obtained with targeted sequence capture.
BACKGROUND
Anaplasma phagocytophilum is a zoonotic and obligate intracellular bacterium transmitted by ticks. In domestic ruminants, it is the causative agent of tick-borne fever, which causes significant economic losses in Europe. As A. phagocytophilum is difficult to isolate and cultivate, only nine genome sequences have been published to date, none of which originate from a bovine strain.Our goals were to; 1/ develop a sequencing methodology which efficiently circumvents the difficulties associated with A. phagocytophilum isolation and culture; 2/ describe the first genome of a bovine strain; and 3/ compare it with available genomes, in order to both explore key genomic features at the species level, and to identify candidate genes that could be specific to bovine strains.


RESULTS
DNA was extracted from a bovine blood sample infected by A. phagocytophilum. Following a whole genome capture approach, A. phagocytophilum DNA was enriched 197-fold in the sample and then sequenced using Illumina technology. In total, 58.9% of obtained reads corresponded to the A. phagocytophilum genome, covering 85.3% of the HZ genome. Then by performing comparisons with nine previously-sequenced A. phagocytophilum genomes, we determined the core genome of these ten strains. Following analysis, 1281 coding DNA sequences, including 1001 complete sequences, were detected in the A. phagocytophilum bovine genome, of which four appeared to be unique to the bovine isolate. These four coding DNA sequences coded for "hypothetical proteins of unknown function" and require further analysis. We also identified nine proteins common to both European domestic ruminants tested.


CONCLUSIONS
Using a whole genome capture approach, we have sequenced the first A. phagocytophilum genome isolated from a cow. To the best of our knowledge, this is the first time that this method has been used to selectively enrich pathogenic bacterial DNA from samples also containing host DNA. The four proteins unique to the A. phagocytophilum bovine genome could be involved in host tropism, therefore their functions need to be explored.
Thibaud
T
Dugat
Valentin
V
Loux
Sylvain
S
Marthey
Marco
M
Moroldo
Anne-Claire
AC
Lagrée
Henri-Jean
HJ
Boulouis
Université Paris-Est, Ecole Nationale Vétérinaire d'Alfort, UMR BIPAR ENVA Anses UPEC USC INRA, Maisons-Alfort, France. nhaddad@vet-alfort.fr.
Nadia
N
Haddad
Renaud
R
Maillard
D041081:Anaplasma phagocytophilum / Q000235:genetics*; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D058947:Bacterial Secretion Systems / Q000235:genetics; D001483:Base Sequence; D002417:Cattle; D002448:Cell Adhesion / Q000235:genetics; D004269:DNA, Bacterial / Q000097:blood / Q000235:genetics; D016873:Ehrlichiosis / Q000097:blood / Q000235:genetics / Q000382:microbiology / Q000662:veterinary; D004705:Endocytosis / Q000235:genetics; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D023281:Genomics / Q000379:methods*; D009504:Neutrophils / Q000378:metabolism; D010802:Phylogeny; D015203:Reproducibility of Results; D055571:Secretory Pathway / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*
D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D004269:DNA, Bacterial