Molecular evidence for Bartonella spp. in cat and dog fleas from Germany and France

Nine hundred and fifty-two fleas were collected from 148 cats and 133 dogs at 18 widely distributed geographic locations in Germany and France and examined for the presence of six different Bartonella spp. (Bartonella bacilliformis, Bartonella clarridgeiae, Bartonella elizabethae, Bartonella henselae, Bartonella quintana, Bartonella vinsonii subsp. berkhoffii) by PCR. Thirty-five specimens (3.7%) tested positive for either B. henselae (14 positive fleas) or B. clarridgeiae (21 positive fleas). DNA of other Bartonella spp. were not detected. Bartonella clarridgeiae was the dominating species in samples from France (19 out of 22 positive fleas), whereas B. henselae was more frequent in Germany (11 out of 13 positive fleas). With 3.5% (22 out of 632 fleas) in France and 4.1% (13 out of 320 fleas) in Germany, the overall prevalences of pathogen did not vary significantly between the flea populations of both countries. 5.4% of cats in France versus 16.1% of cats from Germany were infested by fleas carrying Bartonella, whereas 9.5% of dogs in France but none of the examined dogs from Germany were infested by Bartonella positive fleas. The molecular evidence of Bartonella infections reveals that agents of zoonotic potential are established in flea populations in Germany and France and that the spectrum of species can vary significantly from country to country.     

Bartonella species in wild rodents and fleas from them in Japan

The purpose of this study was to assess the role of fleas for transmission of Bartonella species among wild rodents in Japan. Flea samples were collected from wild rodents and examined genetically for Bartonella infection. Bartonella DNA was detected from 16 of 40 (40.0%) flea samples. Sequence analysis demonstrated that 3 of 16 (18.8%) of the Bartonella-positive animals were infested with fleas from which the closely related Bartonella DNA sequence was detected, indicating that the fleas acquired Bartonella from the infested rodents. The DNA was detected in hemolymph, the midgut and the ovary (only in female), indicating that Bartonella might be colonized through the midgut and distributed into the body.     

Candidatus Bartonella antechini: a novel Bartonella species detected in fleas and ticks from the yellow-footed antechinus (Antechinus flavipes), an Australian marsupial

Bartonella are fastidious, Gram-negative, aerobic bacilli belonging to the Alphaproteobacteria group. In the last ten years, the discovery of new Bartonella species from a variety of mammalian hosts, arthropod vectors and geographical areas has increased. More than 20 species of Bartonella have been identified, of which approximately thirteen are associated with disease in humans and animals. Recently, four novel species of Bartonella were isolated from mammalian hosts in Australia: Bartonella australis from eastern grey kangaroos (Macropus giganteus) and Bartonella rattaustraliani, Bartonella queenslandensis and Bartonella coopersplainsensis from rodents. Bartonella-like organisms have also been detected from Ixodes tasmani ticks collected from koalas (Phascolarctos cinereus). However, very little is known about Bartonella spp. in other marsupials in Australia. We report the identification of a novel Bartonella species detected from fleas (Acanthopsylla jordani) and ticks (Ixodes antechini) collected from a small carnivorous marsupial, Antechinus flavipes (Mardos or Yellow-footed antechinus) in the southwest of Western Australia. New nested-PCRs targeting the gltA gene and the ribosomal ITS region were developed as part of the present study. DNA sequencing of the 16S rRNA, gltA, ftsZ and rpoB genes and the ribosomal ITS region revealed that this detection is a distinct Bartonella species and is related to B. australis isolated from kangaroos. This is the first report of two different possible arthropod vectors in Australia (ticks and fleas) being infected with the same species of Bartonella. We propose the name Candidatus Bartonella antechini n. sp. for the recently characterized organism.     

Diversity of Bartonella species detected in arthropod vectors from animals in Australia

A variety of Bartonella species were detected in two species of ticks and three species of fleas collected from marsupial hosts; brush-tailed bettong or woylie (Bettongia penicillata) and western barred bandicoots (Perameles bougainville) and from a rodent host; Rattus fuscipes in Western Australia. Bartonella species were detected using nested-PCR of the gltA gene and the 16S–23S ribosomal internal transcribed spacer region (ITS), and species were characterized using DNA sequencing of the 16S rRNA, gltA, rpoB, ftsZ genes and the ITS region. Bartonella rattaustraliani and B. coopersplainsensis were detected in Ixodes spp. ticks and fleas (Stephanocircus pectinipes) respectively collected from rodents. Two novel Bartonella species were detected from marsupials; Candidatus Bartonella woyliei n. sp. was detected in both fleas (Pygiopsylla hilli) and ticks (Ixodes australiensis) collected from woylies and Candidatus Bartonella bandicootii n. sp. was detected in fleas (Pygiopsylla tunneyi) collected from western barred bandicoots. Concatenated phylogenetic analysis of all 5 loci clarified the marsupial cluster of Bartonella species in Australia and confirmed the species status of these two Bartonella species in ticks and fleas from woylies and western barred bandicoots, which are classified as threatened species and are vulnerable to extinction.     

Prevalence of Bartonella species DNA and antibodies in cats (Felis catus) submitted to a spay/neuter program in Rio de Janeiro, Brazil

The prevalence of Bartonella species DNA and antibodies for Bartonella henselae were studied in 40 clinically healthy cats (Felis catus, Linnaeus 1758) submitted to a spay/neuter program in Rio de Janeiro, Brazil. Additionally, the prevalence of Bartonella species DNA was investigated in the fleas found parasitizing the subject cats. For this purpose, blood samples were obtained from all cats, and DNA extraction was performed on the blood, and blood clotted samples, as well as on pools of fleas obtained from them. Antibodies for B henselae were detected on serum samples. Bartonella species DNA was detected in 17 cats, whereas serum reactivity for B henselae was found in 19. A total of 20 cats were flea-infested and nine of these 20 had Bartonella species DNA in their blood. In four of the 20 flea-infested cats, Bartonella species DNA was detected in the fleas obtained from those cats, but only one of these four cats had Bartonella species DNA in its blood.     

Rickettsiae in arthropods collected from the North African Hedgehog (Atelerix algirus) and the desert hedgehog (Paraechinus aethiopicus) in Algeria

Hedgehogs have become a popular pet despite their potential role in zoonotic disease transmission. We conducted an entomological study in a mountainous region of northeast Algeria in which we collected 387 fleas (Archeopsylla erinacei) and 342 ticks (Rhipicephalus sanguineus and Haemaphysalis erinacei) from Paraechinus aethiopicus and Atelerix algirus hedgehogs. Of the hedgehogs sampled, 77.7% and 91% were infested with fleas and ticks, respectively. Significantly more ticks and fleas were collected from A. algirus than from P. aethiopicus. Rickettsia felis was detected in 95.5% of fleas and R. massiliae was detected in 6.25% of Rh. sanguineus ticks by molecular tools. A new Rickettsia species of the spotted fever group was detected in 11.25% of Rh. sanguineus and in 77% of H. erinacei ticks. Overall, we show that hedgehogs can act as hosts for ectoparasites infected with several rickettsial agents. These data justify a more detailed investigation of animal reservoirs for Rickettsiae.     

Species of flea (siphonaptera) infesting pets and hedgehogs in Germany

The species of flea infesting pets and hedgehogs in Germany were investigated through a survey of small animal practitioners throughout the country who were asked to collect specimens at their veterinary practices. A total of 625 veterinarians/veterinary practices responded and provided 2445 intact anti identifiable flea specimens. These fleas originated from 294 dogs (795 fleas), 334 cats (1152 fleas), 76 hedgehogs (481 fleas), five domestic rabbits (10 fleas), one golden hamster (four fleas) and one ferret (three fleas). Dogs were found to be infested with Archaeopsylla erinacei, Chaetopsylla globiceps, Ctenocephalides canis, Ctenocephalides felis, Hystrichopsylla talpae, Nosopsyllus fasciatus, Paraceras melis and Pulex irritans. From cats, Archaeopsylla erinacei, Ceratophyllus gallinae, Ceratophyllus garei, Ctenocephalides felis, Ctenophthalmus assimilis, Hystrichopsylla talpae, Monopsyllus sciurorum, Nosopsyllus fasciatus, Spilopsyllus cuniculi and Typhloceras poppei were collected. In both dogs and cats the most prevalent species were Ctenocephalides felis (78.9% and 91.6%, respectively) and Archaeopsylla erinacei (21.1% and 12.6%, respectively) followed by Ctenocephalides canis in dogs (5.8%) and Hystrichopsylla talpae in cats (1.2%). The fleas isolated from rabbits were Ctenocephalides felis, Hystrichopsylla talpae and Spilopsyllus cuniculi. Nosopsyllus fasciatus and Ctenocephalides felis were recovered from the golden hamster and the ferret, respectively. The hedgehogs were found to be infested with Archaeopsylla erinacei, Ceratophyllus gallinae and Ctenocephalides felis.     

Prevalence of Bartonella species, haemoplasma species, Ehrlichia species, Anaplasma phagocytophilum, and Neorickettsia risticii DNA in the blood of cats and their fleas in the United States

Ctenocephalides felis were killed and collected from 92 cats in Alabama, Maryland, and Texas. The fleas and blood from the corresponding cat were digested and assessed in polymerase chain reaction assays that amplify DNA of Ehrlichia species, Anaplasma phagocytophilum, Neorickettsia risticii, Mycoplasma haemofelis, 'Candidatus M haemominutum' and Bartonella species. DNA consistent with B henselae, B clarridgeiae, M haemofelis, or 'Candidatus M haemominutum' was commonly amplified from cats (60.9%) and their fleas (65.2%). Results of this study support the recommendation to maintain flea control on cats in endemic areas.     

Bartonella and Rickettsia in arthropods from the Lao PDR and from Borneo, Malaysia

Rickettsioses and bartonelloses are arthropod-borne diseases of mammals with widespread geographical distributions. Yet their occurrence in specific regions, their association with different vectors and hosts and the infection rate of arthropod-vectors with these agents remain poorly studied in South-east Asia. We conducted entomological field surveys in the Lao PDR (Laos) and Borneo, Malaysia by surveying fleas, ticks, and lice from domestic dogs and collected additional samples from domestic cows and pigs in Laos. Rickettsia felis was detected by real-time PCR with similar overall flea infection rate in Laos (76.6%, 69/90) and Borneo (74.4%, 268/360). Both of the encountered flea vectors Ctenocephalides orientis and Ctenocephalides felis felis were infected with R. felis. The degrees of similarity of partial gltA and ompA genes with recognized species indicate the rickettsia detected in two Boophilus spp. ticks collected from a cow in Laos may be a new species. Isolation and further characterization will be necessary to specify it as a new species. Bartonella clarridgeiae was detected in 3/90 (3.3%) and 2/360 (0.6%) of examined fleas from Laos and Borneo, respectively. Two fleas collected in Laos and one flea collected in Borneo were co-infected with both R. felis and B. clarridgeiae. Further investigations are needed in order to isolate these agents and to determine their epidemiology and aetiological role in unknown fever in patients from these areas.     

Rickettsiae in arthropods collected from red foxes (Vulpes vulpes) in France

The aim of our study was to detect the presence of Rickettsia spp. and Bartonella spp. in ticks and fleas collected from red foxes (Vulpes vulpes) in southeastern France during 2008. Using a genus-specific quantitative PCR (qPCR) assay, which was followed by a species-specific qPCR assay for the positive samples, 45.2% (33/73) of ticks (Rhipicephalus turanicus) were found to be infected with Rickettsia massiliae. 10.5% (2/19) of the fleas (Archaeopsylla erinacei) collected in the study tested positive for Rickettsia felis. A genus-specific qPCR assay did not reveal any Bartonella species in any of the ticks or fleas collected. The role of red foxes in the epidemiology of spotted fever caused by Rickettsiae species requires further investigation.     

Bartonellae of the Namaqua rock mouse, Micaelamys namaquensis (Rodentia: Muridae) from South Africa

The aim of this study was to determine Bartonella prevalence and diversity in Namaqua rock mice, Micaelamys namaquensis, a species endemic to South Africa, which can attain pest status. A total of 100 heart samples collected monthly from March to December were screened for Bartonella genome presence using three primer sets targeting the citrate synthase (gltA) gene, the NADH dehydrogenase gamma subunit (nuoG) gene and the RNA polymerase β-subunit-encoding gene (rpoB). An overall prevalence of 44% was obtained, with no statistically significant differences or correlations between infection rates and rodent sex, month of capture or season of capture. Phylogenetic analysis of 34 unambiguous gltA sequences revealed the presence of three discrete Bartonella lineages in M. namaquensis, one of which corresponds to Bartonella elizabethae, a species with known zoonotic potential.     

Investigation of Bartonella henselae in cats in Ankara, Turkey

The purpose of this study was to determine Bartonella henselae prevalance in cats in Ankara. Whole bloods and sera collected from 256 cats were investigated for the presence feline Bartonella species by culture and sera were tested for the presence of antibodies against B. henselae IgG using immunofluorescence assay. Bartonella species were isolated by blood culture from 24 (9.4%) cats. Bartonella isolates were subjected to restriction fragment length polymorphism (RFLP) by using TaqI and HhaI endonucleases to identify species. Twenty-one isolates were determined as B. henselae and three of 24 isolates were determined as Bartonella clarridgeiae with RFLP. The bacteraemia prevalence and seroprevalence of B. henselae IgG antibodies in cats was detected as 8.2% and 18.6% respectively. This is the first report on B. henselea and B. clarridgeiae in cats in Turkey.     

Identification of novel Bartonella spp. in bats and evidence of Asian gray shrew as a new potential reservoir of Bartonella

Many studies indicated that small mammals are important reservoirs for Bartonella species. Using molecular methods, several studies have documented that bats could harbor Bartonella. This study was conducted to investigate the relationship of Bartonella spp. identified in bats and small mammals living in the same ecological environment. During May 2009 and March 2010, a total of 102 blood specimens were collected. By whole blood culture and molecular identification, a total of 6 bats, 1 rodent and 9 shrews were shown to be infected by Bartonella species. After sequencing and phylogenetic analyses of the sequences of gltA, ftsZ, rpoB and ribC genes, these specific isolates from bats were not similar to the known Bartonella species (the similarity values were less than 91.2%, 90.5%, 88.8%, and 82.2%, respectively); these isolates formed an independent clade away from other known Bartonella type strains. The Bartonella spp. isolated from small mammals, which were closely related to Bartonella tribocorum, Bartonella elizabethae, Bartonella grahamii, Bartonella rattimassiliensis and Bartonella queenslandensis, were similar to the findings in previous studies worldwide. Therefore, the results implied that the species of Bartonella strains isolated from small mammals were different from those identified in bats. Our results strongly suggested that the bat isolate could be a new Bartonella species. This study is also the first one to isolate Bartonella organisms from Asian gray shrews, Crocidura attenuata tanakae.     

Bartonella washoensis infection in red squirrels (Sciurus vulgaris) and their ectoparasites in Lithuania

This is the first study to investigate the presence of Bartonella infections in different internal organs of red squirrels and their ectoparasites in Lithuania. A total of 39 roadkill red squirrels were collected. Squirrels were infested with Ixodes ricinus ticks (191) and Ceratophyllus sciurorum fleas (36). The presence of Bartonella spp. was screened using 16 S–23 S rRNA internal transcribed spacer region and bacteria were detected in 38.5 % (15/39) samples of squirrels, 1.0 % (2/191) samples of ticks and 55.5 % (20/36) samples of fleas. The infection rate of different internal organs of squirrels varied from 11.1%–47.4%. The 16 S–23 S rRNA ITS region sequences showed that Bartonella washoensis were detected in squirrels and their ectoparasites. The results from this study support the hypothesis that S. vulgaris and their fleas, C.sciurorum, serve as a major reservoir and a vector, respectively, of zoonotic B. washoensis in Lithuania.     

Prevalence of DNA of Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum,' Anaplasma phagocytophilum, and species of Bartonella, Neorickettsia, and Ehrlichia in cats used as blood donors in the United States

OBJECTIVE: To identify the prevalence of DNA of Mycoplasma haemofelis; 'Candidatus Mycoplasma haemominutum'; Anaplasma phagocytophilum; and species of Bartonella, Neorickettsia, and Ehrlichia in blood of cats used as blood donors in the United States. DESIGN: Prospective study. ANIMALS: 146 cats that were active blood donors. PROCEDURES: Environmental history was requested for each blood-donor cat from which a blood sample (mixed with EDTA) was available. Polymerase chain reaction assays capable of amplifying the DNA of the microorganisms of interest following DNA extraction from blood were performed. RESULTS: Overall, DNA of one or more of the infectious agents was detected in blood samples from 16 of 146 (11%) feline blood donors. Twenty-eight laboratory-reared cats housed in a teaching hospital had negative results for DNA of all organisms investigated. The DNA of at least 1 infectious agent was amplified from blood samples collected from 16 of 118 (13.6%) community-source cats; assay results were positive for 'Candidatus M haemominutum,' M haemofelis, or Bartonella henselae alone or in various combinations. Of the community-source cats allowed outdoors (n = 61) or with known flea exposure (44), DNA for a hemoplasma or B henselae was detected in 21.3% and 22.7%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: When community-source cats, cats allowed outdoors, or cats exposed to fleas are to be used as blood donors, they should be regularly assessed for infection with M haemofelis, 'Candidatus M haemominutum,' and Bartonella spp, and flea-control treatment should be regularly provided.     

Molecular evidence of vector-borne pathogens in dogs and cats and their ectoparasites in Algiers,Algeria

In Algeria, only limited information is currently available on the prevalence of emergent canine and feline vector-borne diseases. The aim of the present work was to detect by qPCR vector-associated bacteria in stray dogs and cats and their ectoparasites from Algiers.18/117 (15.38%) dogs and 2/107 (1.87%) cats were positive for at least one vector-borne agent. Coxiella burnetii and Bartonella henselae were identified in 1/117 (0.85%) dog individually. Ehrlichia canis DNA was detected in 17/117 (14.52%) dogs. 1/107 (0.93%) cat was positive to C. burnetii and another 1/107 (0.93%) to B. henselae.DNA of Rickettsia massiliae, Rickettsia conorii and E. canis was detected in Rhipicephalus sanguineus. Cat fleas were infected with Rickettsia felis, B. henselae and Bartonella clarridgeiae. B. vinsonii subsp. berkhoffii was identified in Xenopsylla cheopis collected from dogs.The findings of this study indicate that dogs and cats from Algeria are exposed to multiple tick and flea-borne pathogens.     

Detection of Bartonella sp. and a novel spotted fever group Rickettsia sp. in Neotropical fleas of wild rodents (Cricetidae) from Southern Brazil

The Neotropical region shows a great diversity of fleas, comprising more than 50 genera. The importance of the study of fleas is linked to their potential role as disease vectors. The aim of this study is to investigate the presence of Rickettsia spp. and Bartonella spp. in Neotropical fleas collected from wild rodents in Southern Brazil. From 350 rodents captured, 30 were parasitized by fleas. A total of 61 fleas belonging to two genera and six different species were collected (Craneopsylla minerva minerva, Polygenis occidentalis occidentalis, Polygenis platensis, Polygenis pradoi, Polygenis rimatus, and Polygenis roberti roberti). In 13 % of fleas of three different species (C. minerva, P. platensis, and P. pradoi) Rickettsia sp. DNA was found. Phylogenetic analysis of concatenated sequences of gltA, htrA, and ompA genes showed that Rickettsia sp. found in rodent fleas (referred as strain Taim) grouped together with Spotted Fever Group Rickettsia. In reference to Bartonella spp., five genotypes were identified in seven fleas of two species (C. minerva and P. platensis) and in five rodent spleens. Also, 207 frozen samples of wild rodents were screened for these pathogens: while none was positive for Rickettsia spp.; five rodent spleens were PCR-positive for Bartonella spp.. Herein, we show the detection of potential novel variants of Bartonella sp. and Rickettsia sp. in fleas collected of wild rodents from Southern Brazil. Further studies are needed to fully characterize these microorganisms, as well as to improve the knowledge on the potential role of Neotropical flea species as diseases vectors.     

Pathogen carriage by the cat flea Ctenocephalides felis (Bouché) in the United Kingdom

The carriage of Bartonella, Rickettsia felis and haemoplasma species was investigated in cat fleas (Ctenocephalides felis) collected from 121 cats and dogs in the United Kingdom. DNA extracted from fleas was analysed using genus and species-specific PCR and amplicons were characterised using DNA sequencing. Fifty percent of flea samples were PCR positive for at least one pathogen. Twenty one percent were positive for R. felis, 17% for Bartonella henselae, 40% for haemoplasma species and 20% were infected with more than one of the pathogen species studied. It is clear from the results in this study that companion cats and dogs are commonly infested with Ct. felis carrying bacterial pathogens of significance to human and animal health. These findings raise the possibility that Ct. felis found on dogs and cats are a potential source of infection with such pathogens for humans.     

Bartonella DNA in the blood and lymph nodes of Golden Retrievers with lymphoma and in healthy controls

BACKGROUND: Although lymphoma is the most common neoplastic process reported in dogs, its precise etiology is unknown. Golden Retrievers are more likely to develop lymphoma, suggesting a breed predisposition; however, other factors, including environment, immunity, and infection, are likely contributors to oncogenesis. HYPOTHESIS: We hypothesized that the development of lymphoma in Golden Retrievers may be associated with vector-borne infections, specifically Bartonella, Anaplasma, or Ehrlichia species infections. ANIMALS: Golden Retrievers with lymphoma and healthy Golden Retrievers from across the United States were recruited for study participation. METHODS: A matched, case-control study was performed to determine the association of lymphoma and the presence of Bartonella, Anaplasma, and Ehrlichia species in serum, blood, and lymph node aspirates. RESULTS: Using PCR analyses and DNA sequencing, single and coinfections with Bartonella henselae, Bartonella elizabethae, Bartonella quintana, and/or Bartonella vinsonii (berkhoffii) were detected in the blood and lymph node aspirates of Golden Retrievers with lymphoma (5/28 dogs, 18%) and in healthy Golden Retrievers (10/56 dogs, 18%); no Anaplasma or Ehrlichia DNA was detected in any dog. When compared with dogs with lymphoma, a higher (P <.001) proportion of healthy Golden Retrievers were receiving monthly acaricide treatments (2.6 times higher). CONCLUSIONS AND CLINICAL IMPORTANCE: Bartonella DNA can be detected in blood and lymph nodes; importantly, in this report, Bartonella was detected in the same proportion of clinically healthy dogs and dogs with lymphoma. Longitudinal studies should be conducted to determine the mode of transmission of Bartonella in dogs, whether lymphatic infection is persistent, or whether these bacteria may contribute to the development of lymphoma.