Molecular detection of rickettsial agents in ticks and fleas collected from a European hedgehog (Erinaceus europaeus) in Marseilles, France

The European hedgehog (Erinaceus europaeus) is a synanthropic nocturnal insectivore commonly found in the countryside and in the parks and gardens. Because hedgehogs are already involved in the transmission of a number of zoonoses, including salmonellosis and ringworm, we decided to study their possible role in the epidemiology of the spotted fever group of Rickettsia. We collected ticks and fleas from a hedgehog that was captured in the city of Marseilles in France. Using a genus-specific quantitative PCR (qPCR) analysis followed by a species-specific qPCR analysis for positive samples, we observed that 91.7% (11/12) of the Rhipicephalus sanguineus ticks were positive for Rickettsia massiliae and 99.2% (128/129) of the Archaeopsylla erinacei fleas were positive for Rickettsia felis. Hedgehogs carry infected ectoparasites and then likely ensure the dissemination of spotted fever group Rickettsiae, and their epidemiological role requires further investigation.     

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.     

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.     

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.     

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.     

Molecular evidence of vector-borne pathogens coinfecting dogs from Poland

Ticks of the genus Ixodes are vectors for many pathogens, including Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Rickettsia spp., and may also serve as vectors for Bartonella spp. However, the role of ticks in Bartonella transmission requires additional studies. The aim of this study was to investigate whether coinfection with two or more vector-borne pathogens can occur in the following three groups of dogs: I - dogs with suspected borreliosis (N = 92), II - dogs considered healthy (N = 100), and III - dogs with diagnosed babesiosis (N = 50). Polymerase chain reactions were performed to detect DNA of Anaplasma phagocytophilum, Rickettsia spp. and Bartonella spp. in the blood of dogs. In dogs of Group I, the DNA of both A. phagocytophilum and Bartonella sp. was detected (14% and 1%, respectively). In eight dogs, coinfection was indicated: A. phagocytophilum or Bartonella sp. with B. burgdorferi s.l. (the presence of antibodies against and/or DNA B. burgdorferi s.l.). In the case of five dogs positive for A. phagocytophilum DNA, no coinfection with B. burgdorferi s.l. was shown. In Group II, the DNA of A. phagocytophilum was detected in four dogs. In Group III, no pathogenic agents possibly transmitted by ticks were confirmed. No DNA of R. helvetica was detected in any of the groups studied.     

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.     

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.     

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.     

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.     

Borrelia burgdorferi sensu lato and Rickettsia spp. infections in hard ticks (Ixodes ricinus) in the region of Hanover (Germany)

In a total of 605 Ixodes (I.) ricinus ticks collected in the spring-months March, April and May 2005, quantitative real time PCR (qPCR) revealed 26.6% Borrelia (B.) burgdorferi sensu lato (sl)-positive ticks, i. e. divided by sex and stage into 31.9% positive adults (34.8% females and 29.0% males) and 18.5% positive nymphs. Mono-infections with genospecies from the B. burgdorferi sl-complex were found in over two thirds of the positive individuals, whereas almost one third showed double- or even triple-infections. Genospecies-specific conventional PCR determined B. afzelii as the most frequent genospecies followed by B. garinii, B. spielmanii, B. valaisiana and B. burgdorferi sensu stricto (ss). Rickettsia spp. were found in 34.2% of the collected ticks, divided into 37.6% adults (42.5% females and 32.8% males) and 29.0% nymphs. Co-infections of Rickettsia-positive ticks with B. burgdorferi sl spirochaetes were present in 10.1% of the ticks. Thereby, adult ticks exhibited a co-infection rate of 13.4% (15.5% females and 11.3% males) and nymphs of 5.0%. Independently of the above mentioned study, 3939 Ixodes ticks, sent in between 2006 and 2010 for B. burgdorferi sl-diagnostic, were examined by qPCR exclusively for B. burgdorferi sl. The resulting B. burgdorferi sl prevalence was 23.1% and 24.4% in 2006 and 2007, respectively, followed by a continuous decrease to 12.8% in 2010. To analyse whether this observed decrease in infection frequency is due to sampling bias, in a current study randomly sampled ticks collected from defined sites equally distributed over the city of Hanover are investigated in a statistically relevant sample size.     

A serological study of exposure to arthropod-borne pathogens in dogs from northeastern Spain

There is limited information regarding the prevalence of many vector borne pathogens in Europe and especially in Spanish dogs. We investigated 206 sick and 260 clinically healthy dogs from three different regions in northeastern Spain for antibodies to Rickettsia conorii (Rc), Ehrlichia canis (Ec), Anaplasma phagocytophilum (Ap), Bartonella henselae (Bh), Bartonella vinsonii subsp. berkhoffii (Bvb), Leishmania infantum (Li) and Borrelia burgdorferi (Bb) and for antigen of Dirofilaria immitis (Di). Total prevalences were the following: Rc (56.4%), Li (30%), Ec (16.7%), Bh (16.8%), Ap (11.5%), Bvb (1.07%), Di (0.6%) and Bb (0.6%). Seroprevalences for Rc, Ec, Ap, Bh, and Bvb and Bb and Di antigens were similar among the three different study sites. The Ec seroprevalence, as determined by Snap 3DX, was statistically lower in dogs from Mallorca (0%) than Tarragona (16%) and Barcelona (5%) (P < 0.0001). Detection of Rc antibodies was associated with seroreactivity to Ec and Ap antigens (P = 0.018 and P = 0.002, respectively). IFA Ec antibodies were associated with Ap seroreactivity (P < 0.0001). There was no association between the clinical status, sex, time of the year when samples were collected, life-style or exposure to fleas or ticks and a positive test result for Ec, Bh, Bvb, or Bb antibodies or Di antigens. Li seroreactivity was associated with illness and living outdoors (P < 0.0001, P = 0.029; respectively), Rc seroreactivity with the male gender (P = 0.028) and Ap seroreactivity with living outdoors (P = 0.045). This study indicates that exposure to Rc, Li, Ec or related Ehrlichia spp., Bh and Ap or a related spp., is common whereas Di, Bb and Bvb is uncommon among dogs from the Mediterranean basin. We also provide serological data that suggests the existence of a novel Ehrlichia species on Mallorca island.     

Prevalence of Bartonella species, haemoplasmas and Toxoplasma gondii in cats in Scotland

The objective of this study was to determine the prevalence rates for select infectious agents of cats presented to the Royal (Dick) School of Veterinary Studies at the University of Edinburgh, Scotland. Whole blood, serum, and oral mucosal and nail bed swabs were collected. While Ehrlichia species, Anaplasma species or Rickettsia felis DNA were not amplified from any cat, 44.2% of the cats had evidence of infection or exposure to either a Bartonella species (15.3% were seropositive and 5.8% polymerase chain reaction (PCR) positive), a haemoplasma (28.6% PCR positive), and/or Toxoplasma gondii (19.2% seropositive). No Bartonella species DNA was amplified from the nail or oral mucosal swabs despite a 5.8% amplification rate from the blood samples. This finding likely reflects the absence of Ctenocephalides felis infection from our study population, as this organism is a key component for Bartonella species translocation in cats. The results from this study support the use of flea control products to lessen exposure of cats (and people) to Bartonella species and support discouraging the feeding of raw meat to cats and preventing them from hunting to lessen T gondii infection.     

A multi-gene analysis of diversity of bartonella detected in fleas from Algeria

We report the molecular detection of several Bartonella species in 44 (21.5%) of 204 fleas from Algeria collected from 26 rodents and 7 hedgehogs. Bartonella elizabethae and B. clarridgeiae were detected in the fleas collected on hedgehogs. Bartonella tribocorum and B. elizabethae were detected in fleas collected from rats and mice, and sequences similar to an unnamed Bartonella sp. detected in rodents from China were detected in rats as well as a genotype of Bartonella closely related to Bartonella rochalimae detected in fleas collected on brown rats (Rattus norvegicus).     

Investigation of Bartonella infection in ixodid ticks from California

A total of 1253 ixodid ticks (254 tick pools) collected between the end of 1995 and the spring of 1997 from six California counties (El Dorado, Los Angeles, Orange, Santa Cruz, Shasta and Sonoma) were examined for the presence of Bartonella DNA by PCR of the citrate synthase gene. Of 1,119 adult Ixodes pacificus ticks tested, 26 (11.6%) of 224 pools, each containing five ticks, were positive (minimum percentage of ticks harboring detectable Bartonella DNA, 2.3%). Bartonella PCR-positive ticks were identified in five counties but none of the ticks from Los Angeles County was positive. Among 47 nymphal I. pacificus ticks collected in Sonoma County, one (10%) positive pool out of 10 pools was identified (minimum percentage of ticks harboring detectable Bartonella DNA, 2.1%). Among the 54 Dermacentor occidentalis grouped in 12 pools from Orange County, one pool (8.3%) was PCR positive for Bartonella and similarly one pool (14.3%) was positive among the 30 Dermacentor variabilis ticks grouped in seven pools. None of the three D. occidentalis from El Dorado County were positive. None of the nine tick pools positive for Ehrlichia phagocytophila were positive for Bartonella. Following our previous findings of Bartonella PCR-positive adult I. pacificus ticks in central coastal California, this is the first preliminary report of the presence of Bartonella DNA in I. pacificus nymphs and in Dermacentor sp. ticks. Distribution of Bartonella among ixodid ticks appears widespread in California.     

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.     

Flea species infesting dogs in Florida and Bartonella spp. prevalence rates

Several Bartonella spp. associated with fleas can induce a variety of clinical syndromes in both dogs and humans. However, few studies have investigated the prevalence of Bartonella in the blood of dogs and their fleas. The objectives of this study were to determine the genera of fleas infesting shelter dogs in Florida, the prevalence of Bartonella spp. within the fleas, and the prevalence of Bartonella spp. within the blood of healthy dogs from which the fleas were collected. Fleas, serum, and EDTA-anti-coagulated whole blood were collected from 80 healthy dogs, and total DNA was extracted for PCR amplification of Bartonella spp. The genera of fleas infesting 43 of the dogs were determined phenotypically. PCR amplicons from blood and flea pools were sequenced to confirm the Bartonella species. Amplicons for which sequencing revealed homology to Bartonella vinsonii subsp. berkhoffii (Bvb) underwent specific genotyping by targeting the 16S–23S intergenic spacer region.A total of 220 fleas were collected from 80 dogs and pooled by genus (43 dogs) and flea species. Bartonella spp. DNA was amplified from 14 of 80 dog blood samples (17.5%) and from 9 of 80 pooled fleas (11.3%). B. vinsonii subsp. berkhoffii DNA was amplified from nine dogs and five of the flea pools. Bartonella rochalimae (Br) DNA was amplified from six dogs and two flea pools. One of 14 dogs was co-infected with Bvb and Br. The dog was infested with Pulex spp. fleas containing Br DNA and a single Ctenocephalides felis flea. Of the Bvb bacteremic dogs, five and four were infected with genotypes II and I, respectively. Of the Bvb PCR positive flea pools, three were Bvb genotype II and two were Bvb genotype I.Amplification of Bvb DNA from Pulex spp. collected from domestic dogs, suggests that Pulex fleas may be a vector for dogs and a source for zoonotic transfer of this pathogen from dogs to people. The findings of this study provide evidence to support the hypothesis that flea-infested dogs may be a reservoir host for Bvb and Br and that ectoparasite control is an important component of shelter intake protocols.     

Isolation or detection of Bartonella vinsonii subspecies berkhoffii and Bartonella rochalimae in the endangered island foxes (Urocyon littoralis)

Bartonella rochalimae (B.r.) and Bartonella vinsonii subsp. berkhoffii (B.v.b.) have been isolated from gray foxes (Urocyon cinereoargenteus) in mainland California and high Bartonella seroprevalence was reported in island foxes (U. litorralis), especially from Santa Cruz and Santa Rosa Islands. As a follow-up study, the objectives were to determine the prevalence of Bartonella bacteremia and seropositivity and to identify the Bartonella species infecting a convenience sample of 51 island foxes living on Santa Rosa Island. Using an immuno-fluorescence antibody test directed against B.v.b and Bartonella clarridgeiae (B.c.), used as a substitute for B.r., the overall antibody prevalence was 62.7% with 16 (31.4%) foxes seropositive for B.c. only, 5 (9.8%) for B.v.b. only, and 11 (21.6%) for both antigens. B.v.b. was isolated from 6 (11.8%) foxes using blood culture medium. An additional seropositive fox tested PCR positive for B.v.b. and 3 other seropositive foxes tested PCR positive for B. rochalimae. All of the isolated B.v.b. colonies and the B.v.b. PCR positive sample belonged to type III, the same type found to infect mainland gray foxes. Therefore, Bartonella infection is widespread within this island fox population with evidence for B.v.b. type III reservoir host-specificity. Presence of B. rochalimae in the Channel Islands has been detected for the first time using PCR.     

Molecular identification of tick-borne pathogens in Nigerian ticks

A molecular epidemiology investigation was undertaken in two Nigerian states (Plateau and Nassarawa) to determine the prevalence of pathogens of veterinary and public health importance associated with ticks collected from cattle and dogs using PCR, cloning and sequencing or reverse line blot techniques. A total of 218 tick samples, Amblyomma variegatum (N=153), Rhipicephalus (Boophilus) decoloratus (N=45), and Rhipicephalus sanguineus (N=20) were sampled. Pathogens identified in ticks included piroplasmids (Babesia spp., Babesia bigemina and Babesia divergens), Anaplasma marginale and Rickettsia africae. Piroplasmids were identified in A. variegatum, A. marginale was found in R. decoloratus, while R. africae was detected in all tick species examined. Ehrlichia spp. and Theileria spp. were not identified in any of the ticks examined. Of the 218 ticks examined, 33 (15.1%) contained pathogen DNA, with the presence of B. divergens and R. africae that are zoonotic pathogens of public health and veterinary importance. The variety of tick-borne pathogens identified in this study suggests a risk for the emergence of tick-borne diseases in domestic animals and humans, especially amongst the Fulani pastoralists in Plateau and Nassarawa states of Nigeria.     

Prevalence of Bartonella infection in wild African lions (Panthera leo) and cheetahs (Acinonyx jubatus)

Bartonella species are emerging pathogens that have been isolated worldwide from humans and other mammals. Our objective was to estimate the prevalence of Bartonella infection in free-ranging African lions (Panthera leo) and cheetahs (Acinonyx jubatus). Blood and/or serum samples were collected from a convenience sample of 113 lions and 74 cheetahs captured in Africa between 1982 and 2002. Whole blood samples available from 58 of the lions and 17 of the cheetahs were cultured for evidence of Bartonella spp., and whole blood from 54 of the 58 lions and 73 of the 74 cheetahs tested for the presence of Bartonella DNA by TaqMan PCR. Serum samples from the 113 lions and 74 cheetahs were tested for the presence of antibodies against Bartonella henselae using an immunofluorescence assay. Three (5.2%) of the 58 lions and one (5.9%) of the 17 cheetahs were bacteremic. Two lions were infected with B. henselae, based on PCR/RFLP of the citrate synthase gene. The third lion and the cheetah were infected with previously unidentified Bartonella strains. Twenty-three percent of the 73 cheetahs and 3.7% of the 54 lions tested by TaqMan PCR were positive for Bartonella spp. B. henselae antibody prevalence was 17% (19/113) for the lions and 31% (23/74) for the cheetahs. The prevalence of seropositivity, bacteremia, and positive TaqMan PCR was not significantly different between sexes and age categories (juvenile versus adult) for both lions and cheetahs. Domestic cats are thus no longer the only known carriers of Bartonella spp. in Africa. Translocation of B. henselae seronegative and TaqMan PCR negative wild felids might be effective in limiting the spread of Bartonella infection.