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Letter
Bartonella
Species Isolated from Rodents, Greece
Afrodite Tea,* Stella Alexiou-Daniel,†
Androniki Papoutsi,* Anna Papa,* and Antonis Antoniadis*
*Aristotelian University of Thessaloniki, Thessaloniki, Greece; and †American
Hellenic Educational Progressive Association University Hospital, Thessaloniki,
Greece
Suggested citation
for this article:
Tea A, Alexiou-Daniel S, Papoutsi A, Papa A, Antoniadis A. Bartonella
species isolated from rodents, Greece. Emerg Infect Dis [serial online].
2004 May [date cited]. Available from: http://www.cdc.gov/ncidod/EID/vol10no5/03-0430.htm
To the Editor: Domestic cats and human body lice have been identified
as the vectors of Bartonella henselae and B. quintana, respectively,
the primary sources of Bartonella-associated human diseases (1).
Bartonella species are zoonotic agents that have been isolated
from a wide range of mammals in the United States (2)
and Europe (3) and have been associated with human diseases
(4–5).
This study investigated the potential for infection from Bartonella
species in rodents in northern Greece. The small mammals tested were collected
with live traps (6). Two sites were surveyed; the first
was Nevrokopi, a small town in the Rhodope Mountains near the Greek-Bulgarian
border, and the second site included Pramanta, a small village in the
Pindos Mountains, and Matsuki, a small village in northwestern Greece.
At Nevrokopi, 57 small mammals were captured during 887 trap nights for
a success rate of 6.4%. At Pramanta and Matsuki, 13 small mammals were
captured during 400 trap nights for a success rate of 3.3%. The 70 captured
mammals comprised seven species of rodents. Apodemus flavicollis
was the most commonly captured species (87%). Blood samples from each
of the trapped mammals were frozen in liquid nitrogen in the field and
subsequently stored at –70°C before bacteria isolation. Bacteria isolation
was performed as previously described (7). One hundred
microliters of whole mammalian blood was cultured on heart infusion agar
containing 5% rabbit blood (Becton Dickinson, Franklin Lakes, NJ) and
incubated in 5% CO2 at 35°C for a minimum of 4 weeks. DNA
of the putative Bartonella cultures was extracted by using QIAamp
Tissue Kit (Qiagen GmbH, Hilden, Germany). Polymerase chain reaction (PCR)
was performed by using two oligonucleotides specific for the citrate synthase
(gltA) gene of B. henselae Houston 1, primers BhCS 781.p
and BhCS 1137.n. Negative and positive controls (double-distilled H2O
and DNA from cultures of B. henselae) were used in each PCR run.
Products of the correct size were purified (QIAquick PCR Purification
kit, Qiagen GmbH) and sequenced with the same primers, BhCS 781.p and
BhCS1137.n., in both directions, with the Cy5/Cy5.5 Dye Primer Cycle Sequencing
kit on a Long-Read Tower sequencer (Visible Genetics Inc., Toronto, Canada).
Three hundred thirty-eight base-pair sequences of the gltA gene
were obtained and compared with sequences of other known Bartonella
species in GenBank by using the nucleotide BLAST program (National Center
for Biotechnology Information; Available from: www.ncbi.nlm.nih.gov/BLAST/).
Isolates identified as Bartonella species were obtained from 21
of the 70 blood cultures. All were isolated from A. flavicollis,
and one was isolated from Dryomys nitedula. In addition, all were
isolated from the first site (Nevrokopi village), and one was isolated
from the second site (Pramanta village).
Within these 21 Bartonella isolates, eight genotypes were found.
Among these isolates, one (AY435102 isolated from A. flavicollis
trapped in Pramanta), was identical to ma106up strain, isolated from Microtus
agresti (AF391789); another (AY435103 isolated from A. flavicollis
trapped in Nevrokopi), was identical to af82up strain (AF391788), also
isolated from A. flavicollis (3). Both strains
ma106up (AF391789) and af82up (AF391788) were isolated in central Sweden
(3) The rest of Bartonella isolates were from
mammals trapped in Nevrokopi village and were divided into three phylogenetic
groups. The first group, containing 10 isolates (AY435104–AY435113, isolated
from A. flavicollis) and representing four novel genotypes, was
98% similar to B. taylorii (AF191502, isolated from A. sylvaticus)
gltA gene. The second group, consisting of seven isolates that shared
the same genotype (AY435114–AY435120 isolated from A. flavicollis),
was 99% similar to B. birtlesii (AF204272 isolated from Apodemus
spp.). The third group consisted of two isolates that shared the same
genotype (AY435121 isolated from D. nitedula, and AY435122 isolated
from A. flavicollis); this group was 97% similar to B. grahamii
strain V2 (Z70016 isolated from Neomys fodiens).
This is the first study to identify Bartonella in small mammals
in Greece. We found that 31.3% of the examined mammals were infected with
Bartonella spp. The prevalence of culture-positive infections differed
between the two sites (20/57 versus 1/13), although both are mountain
areas with similar environmental and climatic conditions. A high prevalence
of Bartonella infection in small mammals also has been described
in other countries such as the United States (7) and
Sweden (3), where 42.2% and 16.5% of the collected rodents
were infected with Bartonella spp., respectively. As indicated
in these studies, numerous Bartonella species are found in rodents.
A. flavicollis was the most commonly captured species in Sweden
(110/236), as well as in Greece (61/70). Identical Bartonella strains
were isolated from A. flavicollis and Microtus agresti in
both countries. Unlike Sweden, where the most frequent genotype was B.
grahamii, in this study no isolate was identical to any Bartonella
species known to cause human diseases. However, B. elizabethae
was first isolated from a patient with endocarditis, and nothing was known
concerning the organism’s natural history until it was isolated from a
rodent captured in Peru (4).
The occurrence and distribution of Bartonella in European hosts
are largely unknown. Given the existence of Bartonella spp. in
every mammal group examined to date, the diversity of the genus is probably
much greater than has been observed among the strains examined to date.
In Greece, serologic evidence of human infection with B. henselae
and B. quintana (8), has been found and a case
of B. quintana endocarditis has been established (unpub. data).
The public health relevance of Bartonella infections in small mammals
in Greece compared with other countries remains to be defined.
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