Detection of Bartonella quintana (Hyphomicrobiales: Bartonellaceae) Among Day Laborers in Osaka, Japan, 2009-2010.

Bartonella quintana is a gram-negative bacterium causing trench fever, an illness historically acquired by soldiers during World War I. More recently, outbreaks of trench fever have been reported in those experiencing homelessness in the United States, France, Russia, and Tokyo, as well as in children in Nepal and persons in Ethiopia. Reports of B. quintana infection outside of Tokyo are rare in Japan. The aim of this study was to examine body lice and blood obtained from people staying in shelters in Osaka (2009-2010) for B. quintana via polymerase chain reaction and enzyme-linked immunosorbent assays. Day laborers were defined as homeless individuals and shelter residents in this study. We detected genes of B. quintana in body lice by PCR and antibodies against B. quintana. The positive rate of B. quintana genes was 6/10 (60%) in body lice and the seroprevalence (IgG) of B. quintana was 4/10 (40%). This demonstrates that trench fever was endemic in people staying in shelters in Osaka in 2009-2010.

Molecular and serological detection of arthropod-borne pathogens in carnivorous birds from Brazil.

Rickettsiales, Haemosporida and Rhizobiales agents can cause diseases that affect various animal species, including humans. Due to predation behaviour, carnivorous birds may play an important role in spreading these etiological agentes across geographically distant areas, specially if they are migratory. The aim of this study was to investigate the occurrence and to access the phylogenetic relations among Anaplasmataceae (Ehrlichia, Anaplasma, Neorickettsia), Bartonellaceae (Bartonella spp.), and Haemosporida (Plasmodium, Haemoproteus and Leucocytozoon) agents in blood samples from 121 carnivorous birds sampled in the states of São Paulo and Rio de Janeiro, Brazil. Inclusions resembling hemoparasites were not observed in Giemsa-stained preparations. While three animals were seropositive for E. chaffeensis (3.41% [3/88]; 95% CI:1.17-9.55%), five showed antibodies to A. phagocytophilum (5.68% [5/88]; 95% CI: 2.45-12.62%). Despite the detection of rrs gene fragments closely related to E. chaffeensis (4.13% [5/121]; 95% CI: 1.78-9.31%), no positivity was observed in the qPCR based on the genes vlpt for the organism. Similarly, 12 (9.91% [12/121]; 95% CI: 5.76-16.74%) samples were positive in the qPCR for Anaplasma spp. based on groEL gene, but negative in the qPCR for A. phagocytophilum based on msp-2 gene. Three samples were positive in the nPCR for E. canis based on rrs gene. Three samples were positive for Haemoproteus spp. and one for Plasmodium spp. in the nPCR based on cytB gene. Four birds (3.3% [4/121]; 95% CI: 1.29-8.19%) presented co-positivity by Ehrlichia sp. and Anaplasma sp. in molecular assays. One (0.82% [1/121]; 95% CI:0.15-4.53%) bird showed to be seropositive for E. chaffeensis and and positive in PCR for Haemoproteus sp. All birds were negative in the qPCR assay for Bartonella spp. (nuoG). The present work showed the occurrence of Anaplasmataceae agents and hemosporidians in carnivorous birds from southeastern Brazil. The role of these animals in the dispersion of Anaplasmataceae agents should be further investigated.

Genome Evolution of Bartonellaceae Symbionts of Ants at the Opposite Ends of the Trophic Scale.

Many insects rely on bacterial symbionts to supply essential amino acids and vitamins that are deficient in their diets, but metabolic comparisons of closely related gut bacteria in insects with different dietary preferences have not been performed. Here, we demonstrate that herbivorous ants of the genus Dolichoderus from the Peruvian Amazon host bacteria of the family Bartonellaceae, known for establishing chronic or pathogenic infections in mammals. We detected these bacteria in all studied Dolichoderus species, and found that they reside in the midgut wall, that is, the same location as many previously described nutritional endosymbionts of insects. The genomic analysis of four divergent strains infecting different Dolichoderus species revealed genes encoding pathways for nitrogen recycling and biosynthesis of several vitamins and all essential amino acids. In contrast, several biosynthetic pathways have been lost, whereas genes for the import and conversion of histidine and arginine to glutamine have been retained in the genome of a closely related gut bacterium of the carnivorous ant Harpegnathos saltator. The broad biosynthetic repertoire in Bartonellaceae of herbivorous ants resembled that of gut bacteria of honeybees that likewise feed on carbohydrate-rich diets. Taken together, the broad distribution of Bartonellaceae across Dolichoderus ants, their small genome sizes, the specific location within hosts, and the broad biosynthetic capability suggest that these bacteria are nutritional symbionts in herbivorous ants. The results highlight the important role of the host nutritional biology for the genomic evolution of the gut microbiota-and conversely, the importance of the microbiota for the nutrition of hosts.

Death of Military Working Dogs Due to Bartonella vinsonii Subspecies berkhoffii Genotype III Endocarditis and Myocarditis.

OBJECTIVES: As a result of extensive field-related activities, military working dogs (MWDs) have an increased occupational risk for acquiring vector-borne infectious diseases. METHODS: Indirect fluorescent antibody, Bartonella alpha-proteobacteria growth medium enrichment culture, and 16-23S Bartonella intergenic transcribed spacer polymerase chain reaction were performed using blood, serum, or tissue specimens. RESULTS: Endocarditis was diagnosed in three MWDs infected with Bartonella vinsonii subspecies (subsp.) berkhoffii genotype III. One dog was also infected with Bartonella rochalimae. CONCLUSIONS: B. vinsonii subsp. berkhoffii genotype III may represent an occupational risk for MWDs that develop endocarditis or myocarditis. Comprehensive periodic screening for canine vector-borne infections, in particular occult infections caused by Bartonella spp, is prudent to avoid serious or life-threating illnesses.

Absence of zoonotic Bartonella species in questing ticks: first detection of Bartonella clarridgeiae and Rickettsia felis in cat fleas in the Netherlands.

BACKGROUND: Awareness for flea- and tick-borne infections has grown in recent years and the range of microorganisms associated with these ectoparasites is rising. Bartonella henselae, the causative agent of Cat Scratch Disease, and other Bartonella species have been reported in fleas and ticks. The role of Ixodes ricinus ticks in the natural cycle of Bartonella spp. and the transmission of these bacteria to humans is unclear. Rickettsia spp. have also been reported from as well ticks as also from fleas. However, to date no flea-borne Rickettsia spp. were reported from the Netherlands. Here, the presence of Bartonellaceae and Rickettsiae in ectoparasites was investigated using molecular detection and identification on part of the gltA- and 16S rRNA-genes. RESULTS: The zoonotic Bartonella clarridgeiae and Rickettsia felis were detected for the first time in Dutch cat fleas. B. henselae was found in cat fleas and B. schoenbuchensis in ticks and keds feeding on deer. Two Bartonella species, previously identified in rodents, were found in wild mice and their fleas. However, none of these microorganisms were found in 1719 questing Ixodes ricinus ticks. Notably, the gltA gene amplified from DNA lysates of approximately 10% of the questing nymph and adult ticks was similar to that of an uncultured Bartonella-related species found in other hard tick species. The gltA gene of this Bartonella-related species was also detected in questing larvae for which a 16S rRNA gene PCR also tested positive for "Candidatus Midichloria mitochondrii". The gltA-gene of the Bartonella-related species found in I. ricinus may therefore be from this endosymbiont. CONCLUSIONS: We conclude that the risk of acquiring Cat Scratch Disease or a related bartonellosis from questing ticks in the Netherlands is negligible. On the other hand fleas and deer keds are probable vectors for associated Bartonella species between animals and might also transmit Bartonella spp. to humans.

Functional genomics of bacterial pathogens: from post-genomics to therapeutic targets.

A wealth of new data have become available to the scientific community as a result of the sequencing of many pathogen genomes. A recent meeting devoted to functional genomics of pathogenic microorganisms confirmed the notion that bacterial genomes are not static, because large blocks of genes can be acquired or deleted. Less complex environments usually result in reduction in genome size, while genome expansion is usually associated with environmental change and complexity. During the meeting, pathogenicity and evolutionary aspects were illustrated for enteric pathogens, as well as the microevolution of the plague bacillus Yersinia pestis. New clues for evolution and pathogenicity were derived from comparative genomics of Listeria species. The genomic organization of Bartonellae, an emerging human pathogen, was also discussed in an evolutionary context. Population and functional genomics of Anthrax-causing bacteria highlighted current scientific interest in this potential biothreat.

Ectoparasites of gray squirrels in two different habitats and screening of selected ectoparasites for bartonellae.

Gray squirrels, Sciurus carolinensis, were livetrapped in 2 different habitat types, woodland (67 squirrels) and parkland (53 squirrels), in southeastern Georgia. Ectoparasites were recovered from anesthetized squirrels and compared between hosts from the 2 habitats. Because of the absence of low vegetation in parkland habitats, it was hypothesized that the ectoparasite fauna, especially ticks and chiggers, would be more diverse on woodland squirrels. The results were generally in agreement with this hypothesis. Seventeen species of ectoparasites were recovered from woodland squirrels, compared with 6 species from parkland squirrels. Five species of ticks and 3 species of chiggers parasitized the woodland squirrels compared with no ticks or chiggers on the parkland squirrels. Significantly higher infestation prevalences were recorded on woodland compared with parkland squirrels for the flea Orchopeas howardi, the tick Amblyomma americanum, and the mesostigmatid mite Androlaelaps fahrenholzi. The mean intensity for O. howardi also was significantly higher on woodland than on parkland squirrels. Because a new strain of Bartonella sp. was isolated recently from S. carolinensis in Georgia, selected ectoparasites from this study were screened for bartonellae by polymerase chain reaction (PCR). Some of the fleas and lice, but none of the mites tested, were PCR positive, suggesting that fleas, or lice, or both, might be vectors of bartonellae between squirrels. Six distinct strains of Bartonella sp. were detected, 2 in fleas and 4 in lice.

Proposals to unify the genera Grahamella and Bartonella, with descriptions of Bartonella talpae comb. nov., Bartonella peromysci comb. nov., and three new species, Bartonella grahamii sp. nov., Bartonella taylorii sp. nov., and Bartonella doshiae sp. nov.

Polyphasic methods were used to examine the taxonomic positions of three newly identified Grahamella species. A comparison of the 16S rRNA gene sequences of these organisms with the sequences available for other bacteria revealed that these three species form a tight monophyletic cluster with members of the genus Bartonella. This cluster is only remotely related to other members of the order Rickettsiales. Determinations of the levels of DNA relatedness between Grahamella species and Bartonella species (by using a modified hydroxyapatite method) revealed that all of the species belonging to these two genera are distinct but closely related. On the basis of these data and the results of guanine-plus-cytosine content and phenotypic characterization studies, we propose that the genera Grahamella and Bartonella should be unified and that the latter name should be retained. Bartonella talpae and Bartonella peromysci, new combinations for former Grahamella species, are created, and the following three new Bartonella species are described: Bartonella grahamii, Bartonella taylorii, and Bartonella doshiae. A taxonomic analysis of Grahamella species complete the study of all members of the family Bartonellaceae, and the results of this study support the proposal that the family should be transferred out of the order Rickettsiales.

Grahamella in small woodland mammals in the U.K.: isolation, prevalence and host specificity.

Bacteria isolated from the blood of small woodland mammals were identified as members of the genus Grahamella. The prevalence of Grahamella infection among the 37 small mammals examined, detected by cultivation of blood samples, was 62%. This figure is somewhat higher than previous reports. Further characterization of the isolates, based on restriction enzyme analysis of the 16S rRNA gene, serological reactivity and DNA hybridization studies, revealed three distinct Grahamella species. One of the species was found in five different species of small mammal (Apodemus sylvaticus, A. flavicollis, Clethrionomys glareolus, Microtus agrestis and Neomys fodiens). All three species were found in M. agrestis, although there was no evidence of concurrent infection of an animal by more than one species of Grahamella. These observations demonstrate that Grahamella spp. are not host-specific, as previously thought, and that it is therefore invalid to name Grahamella spp. solely on the basis of the host in which they are observed.

Proposals to unify the genera Bartonella and Rochalimaea, with descriptions of Bartonella quintana comb. nov., Bartonella vinsonii comb. nov., Bartonella henselae comb. nov., and Bartonella elizabethae comb. nov., and to remove the family Bartonellaceae from the order Rickettsiales.

DNA hybridization data (hydroxyapatite method, 50 to 70 degrees C) indicate that Rickettsia prowazekii, the type species of the type genus of the family Rickettsiaceae, is substantially less closely related to Rochalimaea species than was previously thought. The levels of relatedness of Rickettsia prowazekii to Rochalimaea species and to Bartonella bacilliformis under optimal conditions for DNA reassociation were 0 to 14%, with 25.5% or greater divergence in related sequences. When stringent reassociation criteria were used, the levels of relatedness were 0 to 2%. The genera Bartonella and Rochalimaea are currently classified in different families (the Bartonellaceae and the Rickettsiaceae) in the order Rickettsiales. On the basis of DNA relatedness data, previous 16S rRNA sequence data, guanine-plus-cytosine contents, and phenotypic characteristics, neither Bartonella bacilliformis nor Rochalimaea species are closely related to other organisms currently classified in the order Rickettsiales. In fact, the closest relative of these organisms is Brucella abortus. It is therefore proposed that the family Bartonellaceae should be removed from the order Rickettsiales. Previous 16S rRNA sequence data and DNA hybridization data revealed high levels of relatedness between Bartonella bacilliformis and the four Rochalimaea species, indicating that these species are members of a single genus. It is proposed that the genus Rochalimaea should be united with the genus Bartonella in the family Bartonellaceae. The name Bartonella is retained as the genus name since it has nomenclatural priority over the name Rochalimaea. This means that new combinations for the Rochalimaea species must be created. Proposals are therefore made for the creation of Bartonella quintana comb. nov., Bartonella vinsonii comb. nov., Bartonella henselae comb. nov., and Bartonella elizabethae comb. nov.

Grahamella legeri sp. n. (Rickettsiales: Bartonellaceae) hemoparasito de Rattus norvegicus (Berkenhout, 1769) / Grahamella legeri sp. n. (Rickettsiales: Bartonellaceae) haemoparasite of Rattus norvegicus (Berkenhout, 1769)

De 106 Rattus norvegicus capturados no Estado do Rio de Janeiro, observou-se que 25,47//estavam parasitados por Grahamella legeri sp. n. na zona rural a prevalência foi de 53,84//e na zona urbana de 16,25//. A riquetsia foi identificada em esfregaços de sangue periférico, corados pelo método Giemsa, sendo analisada morfológica e biologicamente. As tentativas de transmissäo do parasito para diferentes espécies de mamíferos, por inoculaçäo de sangue infectado, só tiveram êxito para R. norvegicus; nestes se provocou aumento da parasitemia conseqüente à esplenectomia ou a injeçöes de dexametasona. Registrou-se a evoluçäo de quadro clínico de anemia induzida pela infecçäo com G. legeri. Näo houve êxito no cultivo da riquetsia em meio de cultura tido como específico para o gênero. Discutiu-se a posiçäo sistemática, concluíndo-se pela impropriedade da denominaçäo de Grahamella muris Leger, 1913, pelo que se propôs o nome Grahamella legeri, em homenagem a André Legér

[Blood parasites of small wild mammals in Steiermark and Burgenland]. / Zur Kenntnis der Blutparasiten wildlebender Kleinsäuger in der Steiermark und im Burgenland.

A total of 488 wild small mammals (16 species) trapped in two federate units of Austria, Steiermark and Burgenland, were examined on the presence of blood parasites. In Steiermark Grahamelles were detected in Neomys fodiens, Clethrionomys glareolus, Microtus arvalis, M. agrestis, Apodemus flavicollis and A. sylvaticus, while Trypanosoma evotomys and Hepatozoon erhardovae were found in C. glareolus, and Babesia microti in Pitymys subterraneus and M. agrestis. In Burgenland Grahamelles were demonstrated in Sorex araneus, C. glareolus, A. sylvaticus, A. flavicollis and Rattus norvegicus, while trypanosoma grosi was encountered in A. flavicollis and Babesia microti in C. glareolus, M. arvalis and A. flavicollis.

Intraerythrocytic parasites in rodent populations of Connecticut: Babesia and Grahamella species.

A total of 612 Peromyscus leucopus, 11 Microtus pennsylvanicus, 21 Clethrionomys gapperi, and 4 Tamias striatus was collected in Connecticut and examined for Babesia and Grahamella during 1976 and 1977. Babesia antibodies were detected in sera of 9 P. leucopus collected from 4 sites. Babesia parasites were not detected in the blood smears of captured rodents. Subsequent splenectomy and subinoculation of blood from these rodents into susceptible animals failed to induce disease and no Babesia was isolated. Six of 10 P. leucopus inoculated with a Shelter Island, New York strain of B. microti remained infected for 3 1/2 months. Indirect fluorescent antibody titers of experimentally infected P. leucopus ranged from 1:8 to 1:256. Prevalence of Grahamella peromysci infection, as determined from examinations of blood smears of P. leucopus, was 13%. This infection rate is a conservative estimate because parasitemia is difficult to detect in intact animals. Twenty of 58 P. leucopus, taken at 2 sites and with negative blood smears for G. peromysci, developed parasitemia after splenectomy.

Haemobartonella canis infection in research dogs.

During 1970-1972 haemobartonellosis occurred in research canines at 2 widely separated institutions. Clinical anemia occurred in a splenectomized dog at a Maryland facility, and subsequent screening disclosed an infection rate of 65% in a group of 20 splenectomized subjects. Treatment was successful, and the animals were used in research. A research institution in Texas encountered a number of dogs with fever (to 106 degrees F) and eosinophilia (to 42%) following minor surgery. Blood from affected animals was injected iv into splenectomized dogs, and 3 of 6 recipients developed haemobartonellosis. Further study was conducted, with some success, to establish a relationship between fever and eosinophilia and Haemobartonella canis infection in nonsplenectomized subjects. Our experiences suggest that haemobartonellosis is a widespread, latent disease of dogs and that significant potential exists for the infection to adversely affect research results.