LETTERS Bartonella Species Germany). Polymerase chain reaction (AY435104–AY435113, isolated from (PCR) was performed by using two A. flavicollis) and representing four Isolated from oligonucleotides specific for the cit- novel genotypes, was 98% similar to Rodents, Greece rate synthase (gltA) gene of B. hense- B. taylorii (AF191502, isolated from lae Houston 1, primers BhCS 781.p A. sylvaticus) gltA gene. The second To the Editor: Domestic cats and and BhCS 1137.n. Negative and posi- group, consisting of seven isolates that human body lice have been identified tive controls (double-distilled H2O shared the same genotype (AY435114- as the vectors of Bartonella henselae and DNA from cultures of B. hense- AY435120 isolated from A. flavicol- and B. quintana, respectively, the pri- lae) were used in each PCR run. lis), was 99% similar to B. birtlesii mary sources of Bartonella-associat- Products of the correct size were puri- (AF204272 isolated from Apodemus ed human diseases (1). Bartonella fied (QIAquick PCR Purification kit, spp.). The third group consisted of two species are zoonotic agents that have Qiagen GmbH) and sequenced with isolates that shared the same genotype been isolated from a wide range of the same primers, BhCS 781.p and (AY435121 isolated from D. nitedula, mammals in the United States (2) and BhCS1137.n., in both directions, with and AY435122 isolated from A. flavi- Europe (3) and have been associated the Cy5/Cy5.5 Dye Primer Cycle collis); this group was 97% similar to with human diseases (4–5). Sequencing kit on a Long-Read B. grahamii strain V2 (Z70016 isolat- This study investigated the poten- Tower sequencer (Visible Genetics ed from Neomys fodiens). tial for infection from Bartonella Inc., Toronto, Canada). Three hundred This is the first study to identify species in rodents in northern Greece. thirty-eight base-pair sequences of the Bartonella in small mammals in The small mammals tested were col- gltA gene were obtained and com- Greece. We found that 31.3% of the lected with live traps (6). Two sites pared with sequences of other known examined mammals were infected were surveyed; the first was Bartonella species in GenBank by with Bartonella spp. The prevalence Nevrokopi, a small town in the using the nucleotide BLAST program of culture-positive infections differed Rhodope Mountains near the Greek- (National Center for Biotechnology between the two sites (20/57 versus Bulgarian border, and the second site Information; Available from: 1/13), although both are mountain included Pramanta, a small village in www.ncbi.nlm.nih.gov/BLAST/). areas with similar environmental and the Pindos Mountains, and Matsuki, a Isolates identified as Bartonella climatic conditions. A high prevalence small village in northwestern Greece. species were obtained from 21 of the of Bartonella infection in small mam- At Nevrokopi, 57 small mammals 70 blood cultures. All were isolated mals also has been described in other were captured during 887 trap nights from A. flavicollis, and one was isolat- countries such as the United States (7) for a success rate of 6.4%. At ed from Dryomys nitedula. In addi- and Sweden (3), where 42.2% and Pramanta and Matsuki, 13 small tion, all were isolated from the first 16.5% of the collected rodents were mammals were captured during 400 site (Nevrokopi village), and one was infected with Bartonella spp., respec- trap nights for a success rate of 3.3%. isolated from the second site tively. As indicated in these studies, The 70 captured mammals comprised (Pramanta village). numerous Bartonella species are seven species of rodents. Apodemus Within these 21 Bartonella iso- found in rodents. A. flavicollis was the flavicollis was the most commonly lates, eight genotypes were found. most commonly captured species in captured species (87%). Blood sam- Among these isolates, one (AY435102 Sweden (110/236), as well as in ples from each of the trapped mam- isolated from A. flavicollis trapped in Greece (61/70). Identical Bartonella mals were frozen in liquid nitrogen in Pramanta), was identical to ma106up strains were isolated from A. flavicol- the field and subsequently stored at strain, isolated from Microtus agresti lis and Microtus agresti in both coun- –70ºC before bacteria isolation. (AF391789); another (AY435103 iso- tries. Unlike Sweden, where the most Bacteria isolation was performed as lated from A. flavicollis trapped in frequent genotype was B. grahamii, in previously described (7). One hun- Nevrokopi), was identical to af82up this study no isolate was identical to dred microliters of whole mammalian strain (AF391788), also isolated from any Bartonella species known to blood was cultured on heart infusion A. flavicollis (3). Both strains cause human diseases. However, B. agar containing 5% rabbit blood ma106up (AF391789) and af82up elizabethae was first isolated from a (Becton Dickinson, Franklin Lakes, (AF391788) were isolated in central patient with endocarditis, and nothing NJ) and incubated in 5% CO at 35°C 2 Sweden (3) The rest of Bartonella iso- was known concerning the organism’s for a minimum of 4 weeks. DNA of lates were from mammals trapped in natural history until it was isolated the putative Bartonella cultures was Nevrokopi village and were divided from a rodent captured in Peru (4). extracted by using QIAamp Tissue into three phylogenetic groups. The The occurrence and distribution of Kit (Qiagen GmbH, Hilden, first group, containing 10 isolates Bartonella in European hosts are Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 10, No. 5, May 2004 963 LETTERS largely unknown. Given the existence of Bartonella in rodents from the southeast- ly (9). Thus, the possibility of epidem- of Bartonella spp. in every mammal ern United States. Am J Trop Med ic typhus could not be excluded defin- Hyg1997;57:578–88. group examined to date, the diversity 8. Tea A, Alexiou-Daniel S, Arvanitidou M, itively in these cases. of the genus is probably much greater Diza E, Antoniadis A. Occurrence of On May 4, 2003, a 56-year-old than has been observed among the Bartonella henselae and Bartonella quin- man living in Tokushima, Japan, strains examined to date. In Greece, tana in a healthy Greek population. Am J sought medical care; he had a temper- serologic evidence of human infection Trop Med Hyg 2003;68:554–6. ature of 39.1°C and exanthema on the with B. henselae and B. quintana (8), trunk and the upper limbs. No surface Address for correspondence: Stella Alexiou- has been found and a case of B. quin- lymph nodes were palpable. He was Daniel, Department of Microbiology, AHEPA tana endocarditis has been established treated with lincomycin and cefdi- University Hospital, Thessaloniki 54006, (unpub. data). The public health rele- toren pivoxil with no improvement. Greece; fax: +32310994821; email: salex- vance of Bartonella infections in On day 3, the patient informed care- [email protected] small mammals in Greece compared givers that he had been in a bamboo with other countries remains to be grove on days 1 and 11 before the defined. onset of symptoms. C-reactive protein of the serum sample collected on day Afrodite Tea,* 3 was positive (= 7.6 mg/dL). From Stella Alexiou-Daniel,† Reemerging Murine this finding, spotted fever was sus- Androniki Papoutsi,* Anna Papa,* Typhus, Japan pected; the disease is endemic in and Antonis Antoniadis* Tokushima. On day 4, the exanthema *Aristotelian University of Thessaloniki, To the Editor: Murine typhus is had spread systemically, and treat- Thessaloniki, Greece; and †American an arthropod-borne infectious disease ment with minocycline was started, Hellenic Educational Progressive caused by Rickettsia typhi, which is which led to a gradual decrease in Association University Hospital, distributed widely around the world Thessaloniki, Greece fever and rashes. The patient was (1–4). In Japan, tsutsugamushi dis- admitted to the Tokushima University ease occurs most frequently in per- References Hospital on day 6 of the illness for sons infected with rickettsioses (5). diagnosis and further treatment. 1. Karem KL, Paddock CD, Regnery RL. Spotted fever caused by R. japonica Serum samples were collected Bartonella henselae, Bartonella quintana also occurs in the southwestern part of and Bartonella bacilliformis: historical from the patient on days 5, 6, 9, 20, pathogens of emerging significance. Japan (6,7). In the 1940s and 1950s, and 34. Indirect immunoperoxidase Microbes Infect 2000;2:1193–205. many murine typhus cases were tests on the serum samples for tsut- 2. Ellis BA, Regnery RL, Beati L, Bacellar F, reported in Japan. These diagnoses sugamushi disease, spotted fever, Rood M, Glass G, et al. Rats of the genus were made according to the clinical Rattus are reservoir hosts for pathogenic murine typhus, and Q fever on day 5 Bartonella species: an Old World origin for features of the illness and the reactiv- of the illness were negative for a New World disease? J Infect Dis ity of the serum samples to OX19 in immunoglobulin (Ig) G and IgM anti- 1999;180:220–4. Weil-Felix tests. A few cases were bodies (<1:40). Weil-Felix tests on the 3. Holmberg M, Mills JN, McGill S, diagnosed on the basis of symptoms Benjamin G, Ellis BA. Bartonella infection serum samples on days 5 and 9 of the in sylvatic small mammals of central exhibited by animals infected with illness were negative for OX2, OX19, Sweden. Epidemiol Infect 2003;130: isolated rickettsiae and complement and OXK. Indirect immunofluores- 149–57. fixation tests, in addition to results of cence of the serum samples on days 6, 4. Birtles RJ, Raoult D. Comparison of partial the Weil-Felix tests. The Weil-Felix citrate synthase gene (gltA) sequences for 9, 20, and 34 of the illness was con- phylogenetic analysis of Bartonella test is useful for preliminary screen- ducted by using strains 18 and species.