Jennifer B. Henn

Bartonella endocarditis: A pathology shared by animal reservoirs and patients

Bartonellae were first recognized to cause endocarditis in humans in 1993 when cases caused by Bartonella quintana, B. elizabethae, and B. henselae were reported. Since the first isolation of Bartonella vinsonii subspecies berkhoffii from a dog with endocarditis, this organism has emerged as an important pathogen in dogs and an emerging pathogen in people. Subsequently, four types of B. vinsonii subsp. berkhoffii have been described, all of which have been associated with endocarditis in dogs. A limited number of dog endocarditis cases have also been associated with B. clarridgeiae, B. washoensis, B. quintana, and B. rochalimae. The second canine B. clarridgeiae endocarditis case is presented. The clinical and pathological characteristics of Bartonella endocarditis in dogs are similar to disease observed in humans, more often affecting the aortic valve, presenting with highly vegetative lesions with accompanying calcification, and in most instances high antibody titers. Pathological features in dogs include a combination of fibrosis, mineralization, endothelial proliferation, and neovascularization with variable inflammation. Endocarditis has also been described in animal species, which are the natural reservoir of specific Bartonella species, once thought to be solely healthy carriers of these pathogens. A few Bartonella endocarditis cases, including B. henselae, have been reported in cats in the USA and Australia. The second case of B. henselae type Houston I identified in the USA is presented. Furthermore, two cases of B. bovis endocarditis were recently described in adult cows from France. Finally, on‐going investigation of valvular endocarditis in free‐ranging Alaskan sea otters suggests the involvement of Bartonella species.

Bartonella infection in domestic cats and wild felids.

Abstract:  Bartonella are vector‐borne, fastidious Gram‐negative bacteria causing persistent bacteremia in their reservoir hosts. Felids represent a major reservoir for several Bartonella species. Domestic cats are the main reservoir of B. henselae, the agent of cat‐scratch disease. Prevalence of infection is highest in warm and humid climates that are optimal for the survival of cat fleas, as fleas are essential for the transmission of the infection. Flea feces are the likely infectious substrate. Prevalence of B. henselae genotypes among cat populations varies worldwide. Genotype Houston I is more prevalent in the Far East and genotype Marseille is dominant in western Europe, Australia, and the western United States. Cats are usually asymptomatic, but uveitis, endocarditis, neurological signs, fever, necrotic lesions at the inoculation site, lymphadenopathy, and reproductive disorders have been reported in naturally or experimentally infected cats. Domestic cats are also the reservoir of B. clarridgeiae and co‐infection has been demonstrated. B. koehlerae has been isolated from domestic cats, and was identified in cat fleas and associated with a human endocarditis case. B. bovis was isolated from a few cats in the United States and B. quintana DNA was recently identified in a cat tooth. Bartonella spp. have also been isolated from free‐ranging and captive wild felids from North America and Africa. Whereas, B. henselae was identified in African lions and a cheetah, some strains specific to these wild cats have also been identified, leading to the concept of a B. henselae group including various subspecies, as previously described for B. vinsonii.

Bartonella rochalimae in Raccoons, Coyotes, and Red Foxes

To determine additional reservoirs for Bartonella rochalimae, we examined samples from several wildlife species. We isolated B. rochalimae from 1 red fox near Paris, France, and from 11 raccoons and 2 coyotes from California, USA. Co-infection with B. vinsonii subsp. berkhoffii was documented in 1 of the coyotes.

Infective Endocarditis in a Dog and the Phylogenetic Relationship of the Associated “Bartonella rochalimae” Strain with Isolates from Dogs, Gray Foxes, and a Human

The first case of canine endocarditis caused by “Bartonella rochalimae” is reported. By PCR-restriction fragment length polymorphism, sequence, and phylogenetic analyses, Bartonella isolates from a dog with endocarditis, 22 gray foxes, and three dogs, described as B. clarridgeiae like, were confirmed to belong to the new species “B. rochalimae,” suggesting canids as the natural reservoir.

Zoonotic Bartonella Species in Fleas Collected on Gray Foxes (Urocyon cinereoargenteus)

Bartonella spp. are fastidious, gram-negative, rod-shaped bacteria and are usually vector-borne. However, the vector has not been definitively identified for many recently described species. In northern California, gray foxes (Urocyon cinereoargenteus) are infected with two zoonotic Bartonella species, B. rochalimae and B. vinsonii subsp. berkhoffii. Fleas (range 1-8 fleas per fox) were collected from 22 (41.5%) of 54 gray foxes from urban and backcountry zones near Hoopa, California. The flea species were determined, and DNA was individually extracted to establish the Bartonella species harbored by these fleas. Of the 108 fleas collected, 99 (92%) were identified as Pulex simulans. Overall, 39% (42/108) of the fleas were polymerase chain reaction (PCR)-positive for Bartonella, with B. rochalimae and B. vinsonii subsp. berkhoffii identified in 34 (81%) and 8 (19%) of the PCR-positive fleas, respectively. There was no difference between the prevalence of Bartonella spp. in P. simulans for the urban and backcountry zones. Fourteen (64%) of the 22 foxes were Bartonella bacteremic at one or more of the capture dates. In 10 instances, both the foxes and the fleas collected from them at the same blood collection were Bartonella-positive. B. rochalimae was the predominant species identified in both foxes and fleas. The competency of Pulex fleas as a vector of B. rochalimae has not been confirmed and will need to be demonstrated experimentally. Pulex spp. fleas readily feed on humans and may represent a source of human exposure to zoonotic species of Bartonella.

Spatial analysis of the exposure of dogs in rural north-coastal California to vectorborne pathogens

Between 0 and 50 per cent of the dogs in eight rural villages in far northern California with a high risk of tickborne diseases were seropositive for Anaplasma phagocytophilum and Bartonella vinsonii subspecies berkhoffii, and between 0 and 10 per cent were seropositive for Borrelia burgdorferi. The odds ratio for the co-exposure of individual dogs to B vinsonii berkhoffii and A phagocytophilum was 18·2. None of the diseases was associated with the sex of the dogs, whether they slept out of doors, or whether tick-preventive measures were taken. When the villages were assessed for landscape risk factors, a particularly high seroprevalence for B vinsonii berkhoffii and A phagocytophilum was observed in a village at a relatively high altitude and greater distance from the Pacific coast, and montane hardwood conifer woodland was most associated with a high seroprevalence for these two pathogens.

Dogs are more permissive than cats or guinea pigs to experimental infection with a human isolate of Bartonella rochalimae

Bartonella rochalimae was first isolated from the blood of a human who traveled to Peru and was exposed to multiple insect bites. Foxes and dogs are likely natural reservoirs for this bacterium. We report the results of experimental inoculation of two dogs, five cats and six guinea pigs with the only human isolate of this new Bartonella species. Both dogs became bacteremic for 5–7 weeks, with a peak of 103–104 colony forming units (CFU)/mL blood. Three cats had low bacteremia levels (< 200 CFU/mL) of 6–8 weeks’ duration. One cat that remained seronegative had two bacterial colonies isolated at a single culture time point. A fifth cat never became bacteremic, but seroconverted. None of the guinea pigs became bacteremic, but five seroconverted. These results suggest that dogs could be a reservoir of this strain of B. rochalimae, in contrast to cats and guinea pigs.

Seroprevalence of Bartonella spp. in the endangered island fox (Urocyon littoralis)

Bartonella clarridgeiae-like strains, presently B. rochalimae, were isolated in gray foxes (Urocyon cinereoargenteus) in mainland California. The objective of this study was to investigate the presence of Bartonella infection in the endangered island fox (Urocyon littoralis) found only on the Channel Islands off the Californian coast. Between 2001 and 2004, 263 serum samples were collected. Antibodies against Bartonella vinsonii subsp. berkhoffii (Bvb) and B. clarridgeiae (Bc) were detected using an immuno-fluorescence antibody test. Sixty-eight (25.8%) and 73 (27.7%) foxes were positive for Bvb and Bc, respectively. Seroprevalence was the highest on Santa Cruz Island (n=36, Bvb=80.5%; Bc=86.1%) and Santa Rosa Island (n=38, Bvb=52.6%; Bc=65.8%). On San Miguel and San Clemente Islands, seroprevalence for Bvb was 20% and 17.3% respectively, and 0% and 21.3% for Bc. Prevalence ranged between 0% and 5.1% on San Nicolas and Santa Catalina Islands. Foxes from Santa Rosa and Santa Cruz Islands were 17.5 times and 31.5 times as likely to be seropositive for Bvb and Bc than foxes from the other islands (95% confidence interval [95% CI]=8.5, 36.7; 14.4, 70.2). There were no statistically significant differences for presence of Bartonella antibodies by sex, age, origin (captive vs. wild) or year of blood collection. This is the first report of exposure to Bartonella in the island fox population. Further studies are necessary to isolate these bacteria from foxes and determine factors associated with presence or absence of Bartonella species on specific islands.

Experimental infection of dogs with various Bartonella species or subspecies isolated from their natural reservoir

Dogs can be infected by a wide variety of Bartonella species. However, limited data is available on experimental infection of dogs with Bartonella strains isolated from domestic animals or wildlife. We report the inoculation of six dogs with Bartonella henselae (feline strain 94022, 16S rRNA type II) in three sets of two dogs, each receiving a different inoculum dose), four dogs inoculated with B. vinsonii subsp. berkhoffii type I (ATCC strain, one mongrel dog) or type II (coyote strain, two beagles and one mongrel) and B. rochalimae (coyote strain, two beagles). None of the dogs inoculated with B. henselae became bacteremic, as detected by classical blood culture. However, several dogs developed severe necrotic lesions at the inoculation site and all six dogs seroconverted within one to two weeks. All dogs inoculated with the B. v. berkhoffii and B. rochalimae strains became bacteremic at levels comparable to previous experimental infections with either a dog isolate or a human isolate. Our data support that dogs are likely accidental hosts for B. henselae, just like humans, and are efficient reservoirs for both B. v. berkhoffii and B. rochalimae.

Experimental infection of cats with Afipia felis and various Bartonella species or subspecies.

Based upon prior studies, domestic cats have been shown to be the natural reservoir for Bartonella henselae, Bartonella clarridgeiae and Bartonella koehlerae. However, other Bartonella species, such as Bartonella vinsonii subsp. berkhoffii, Bartonella quintana or Bartonella bovis (ex weissii) have been either isolated from or Bartonella DNA sequences PCR amplified and sequenced. In the late 1980s, before B. henselae was confirmed as the etiological agent of cat scratch disease, Afipia felis had been proposed as the causative agent. In order to determine the feline susceptibility to A. felis, B. vinsonii subsp. berkhoffii, Bartonella rochalimae, B. quintana or B. bovis, we sought to detect the presence of bacteremia and seroconversion in experimentally-inoculated cats. Most of the cats seroconverted, but only the cats inoculated with B. rochalimae became bacteremic, indicating that cats are not natural hosts of A. felis or the other Bartonella species or subspecies tested in this study.