Here, we report the culture and characterization of an alphaproteobacterium of the order Rhizobiales, isolated from the gut of the honey bee Apis mellifera. Strain PEB0122T shares >95 % 16S rRNA gene sequence similarity with species of the genus Bartonella, a group of mammalian pathogens transmitted by bloodsucking arthropods. Phylogenetic analyses showed that PEB0122T and related strains from the honey bee gut form a sister clade of the genus Bartonella. Optimal growth of strain PEB0122T was obtained on solid media supplemented with defibrinated sheep blood under microaerophilic conditions at 35–37 °C, which is consistent with the cultural characteristics of other species of the genus Bartonella. Reduced growth of strain PEB0122T also occurred under aerobic conditions. The rod-shaped cells of strain PEB0122T had a mean length of 1.2–1.8 μm and revealed hairy surface structures. Strain PEB0122T was positive for catalase, cytochrome c oxidase, urease and nitrate reductase. The fatty acid composition was comparable to those of other species of the genus Bartonella, with palmitic acid (C16 : 0) and isomers of 18- and 19-carbon chains being the most abundant. The genomic DNA G+C content of PEB0122T was determined to be about 45.5 mol%. The high 16S rRNA gene sequence similarity with species of Bartonella and its close phylogenetic position suggest that strain PEB0122T represents a novel species within the genus Bartonella, for which we propose the name Bartonella apis sp. nov. The type strain is PEB0122T ( = NCIMB 14961T = DSM 29779T).
AhnJ.-H., HongI.-P., BokJ.-I., KimB.-Y., SongJ., WeonH.-Y.2012; Pyrosequencing analysis of the bacterial communities in the guts of honey bees Apis cerana and
Apis mellifera in Korea. J Microbiol 50:735–745 [View Article][PubMed]
BabendreierD., JollerD., RomeisJ., BiglerF., WidmerF.2007; Bacterial community structures in honeybee intestines and their response to two insecticidal proteins. FEMS Microbiol Ecol 59:600–610 [View Article][PubMed]
CariveauD. P., PowellJ. E., KochH., WinfreeR., MoranN. A.2014; Variation in gut microbial communities and its association with pathogen infection in wild bumble bees (Bombus). ISME J 8:2369–2379 [View Article][PubMed]
CiccarelliF. D., DoerksT., von MeringC., CreeveyC. J., SnelB., BorkP.2006; Toward automatic reconstruction of a highly resolved tree of life. Science 311:1283–1287 [View Article][PubMed]
ClarridgeJ. E.III, RaichT. J., PirwaniD., SimonB., TsaiL., Rodriguez-BarradasM. C., RegneryR., ZolloA., JonesD. C., RamboC.1995; Strategy to detect and identify Bartonella species in routine clinical laboratory yields Bartonella henselae from human immunodeficiency virus-positive patient and unique Bartonella strain from his cat. J Clin Microbiol 33:2107–2113[PubMed]
Corby-HarrisV., SnyderL. A., SchwanM. R., MaesP., McFrederickQ. S., AndersonK. E.2014; Origin and effect of Alpha 2.2 Acetobacteraceae in honey bee larvae and description of Parasaccharibacter apium gen. nov., sp. nov. Appl Environ Microbiol 80:7460–7472 [View Article][PubMed]
EllegaardK. M., TamaritD., JavelindE., OlofssonT. C., AnderssonS. G., VásquezA.2015; Extensive intra-phylotype diversity in lactobacilli and bifidobacteria from the honeybee gut. BMC Genomics 16:284 [View Article][PubMed]
EngelP., MartinsonV. G., MoranN. A.2012; Functional diversity within the simple gut microbiota of the honey bee. Proc Natl Acad Sci U S A 109:11002–11007 [View Article][PubMed]
EngelP., KwongW. K., MoranN. A.2013; Frischella perrara gen. nov., sp. nov., a gammaproteobacterium isolated from the gut of the honeybee, Apis mellifera
. Int J Syst Evol Microbiol 63:3646–3651 [View Article][PubMed]
GuindonS., DufayardJ.-F., LefortV., AnisimovaM., HordijkW., GascuelO.2010; New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 59:307–321 [View Article][PubMed]
KearseM., MoirR., WilsonA., Stones-HavasS., CheungM., SturrockS., BuxtonS., CooperA., MarkowitzS., other authors. 2012; Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28:1647–1649 [View Article][PubMed]
KillerJ., DubnáS., SedláčekI., ŠvecP.2014; Lactobacillus apis sp. nov., from the stomach of honeybees (Apis mellifera), having an in vitro inhibitory effect on the causative agents of American and European foulbrood. Int J Syst Evol Microbiol 64:152–157 [View Article][PubMed]
KosoyM., MorwayC., SheffK. W., BaiY., ColbornJ., ChalcraftL., DowellS. F., PeruskiL. F., MaloneyS. A., other authors. 2008; Bartonella tamiae sp. nov., a newly recognized pathogen isolated from three human patients from Thailand. J Clin Microbiol 46:772–775 [View Article][PubMed]
KwongW. K., MoranN. A.2013; Cultivation and characterization of the gut symbionts of honey bees and bumble bees: description of Snodgrassella alvi gen. nov., sp. nov., a member of the family Neisseriaceae of the Betaproteobacteria, and Gilliamella apicola gen. nov., sp. nov., a member of Orbaceae fam. nov., Orbales ord. nov., a sister taxon to the order ‘Enterobacteriales’ of the Gammaproteobacteria
. Int J Syst Evol Microbiol 63:2008–2018 [View Article][PubMed]
KwongW. K., EngelP., KochH., MoranN. A.2014a; Genomics and host specialization of honey bee and bumble bee gut symbionts. Proc Natl Acad Sci U S A 111:11509–11514 [View Article][PubMed]
KwongW. K., MancenidoA. L., MoranN. A.2014b; Genome sequences of Lactobacillus sp. strains wkB8 and wkB10, members of the Firm-5 clade, from honey bee guts. Genome Announc 2:e01176–e14 [View Article][PubMed]
La ScolaB., ZeaiterZ., KhamisA., RaoultD.2003; Gene-sequence-based criteria for species definition in bacteriology: the Bartonella paradigm. Trends Microbiol 11:318–321 [View Article][PubMed]
MoranN. A., HansenA. K., PowellJ. E., SabreeZ. L.2012; Distinctive gut microbiota of honey bees assessed using deep sampling from individual worker bees. PLoS One 7:e36393 [View Article][PubMed]
RussellJ. A., MoreauC. S., Goldman-HuertasB., FujiwaraM., LohmanD. J., PierceN. E.2009; Bacterial gut symbionts are tightly linked with the evolution of herbivory in ants. Proc Natl Acad Sci U S A 106:21236–21241 [View Article][PubMed]
SabreeZ. L., HansenA. K., MoranN. A.2012; Independent studies using deep sequencing resolve the same set of core bacterial species dominating gut communities of honey bees. PLoS One7e41250 [View Article][PubMed]
ThompsonJ. D., HigginsD. G., GibsonT. J.1994; clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680 [View Article][PubMed]