In addition to inducing life-threatening illnesses, such as endocarditis, myocarditis, and meningoencephalitis and contributing to chronic debilitating disease, such as arthritis, osteomyelitis, and granulomatous inflammation in cats, dogs, and potentially other animal species; pets and wildlife species can serve as persistently infected reservoir hosts for the transmission of Bartonella spp. infection to veterinary professionals and others with direct animal contact.

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Using PCR in conjunction with pre-enrichment culture, we detected Bartonella henselae and B. vinsonii subspecies berkhoffii in the blood of 14 immunocompetent persons who had frequent animal contact and arthropod exposure.

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Background: Molecular diagnosis of canine bartonellosis can be extremely challenging and often requires the use of an enrichment culture approach followed by PCR amplification of bacterial DNA.Hypotheses: (1) The use of enrichment culture with PCR will increase molecular detection of bacteremia and will expand the diversity of Bartonella species detected. (2) Serological testing for Bartonella henselae and Bartonella vinsonii subsp. berkhoffii does not correlate with documentation of bacteremia.Animals: Between 2003 and 2009, 924 samples from 663 dogs were submitted to the North Carolina State University, College of Veterinary Medicine, Vector Borne Diseases Diagnostic Laboratory for diagnostic testing with the Bartonella a-Proteobacteria growth medium (BAPGM) platform. Test results and medical records of those dogs were retrospectively reviewed.Methods: PCR amplification of Bartonella sp. DNA after extraction from patient samples was compared with PCR after BAPGM enrichment culture. Indirect immunofluorescent antibody assays, used to detect B. henselae and B. vinsonii subsp. berkhoffii antibodies, were compared with PCR.Results: Sixty-one of 663 dogs were culture positive or had Bartonella DNA detected by PCR, including B. henselae (30/61), B. vinsonii subsp. berkhoffii (17/61), Bartonella koehlerae (7/61), Bartonella volans-like (2/61), and Bartonella bovis (2/61). Coinfection with more than 1 Bartonella sp. was documented in 9/61 dogs. BAPGM culture was required for PCR detection in 32/61 cases. Only 7/19 and 4/10 infected dogs tested by IFA were B. henselae and B. vinsonii subsp. berkhoffii seroreactive, respectively.Conclusions and Clinical Importance: Dogs were most often infected with B. henselae or B. vinsonii subsp. berkhoffii based on PCR and enrichment culture, coinfection was documented, and various Bartonella species were identified. Most infected dogs did not have detectable Bartonella antibodies.

Bacteria of the genus Bartonella, a member of the Alphaproteobacteria, are fastidious, gram-negative, aerobic bacilli that comprise numerous species, subspecies, and subtypes. In human and veterinary medicine, species isolation remains a vital component of the diagnostic and therapeutic management of Bartonella infection. We describe a novel, chemically modified, insect-based liquid culture medium that supports the growth of at least seven Bartonella species. This medium will also support cocultures consisting of different Bartonella species, and it facilitated the primary isolation of Bartonella henselae from blood and aqueous fluid of naturally infected cats. This liquid growth medium may provide an advantage over conventional direct blood agar plating for the diagnostic confirmation of bartonellosis.Due to their zoonotic potential, their vector transmission, which includes sandflies, lice, fleas, and ticks, and their frequent adaptation to a mammalian reservoir host, Bartonella species are considered among the newest and most significant emerging pathogens (1,3,8,12,27,29,39,45). These bacteria are highly adapted to a mammalian reservoir host; further, these organisms have been shown to cause a long-lasting intraerythrocytic bacteremia in both humans and animals (11,14,22,24,25,37). Bartonella species are also the causative agents of Carrion's disease (Oroya fever and verruga peruana) (Bartonella bacilliformis) (6), trench fever (B. quintana) (11,40,43), endocarditis (B. elizabethae, B. henselae, B. vinsonii subsp. berkhoffii, B. washoensis, B. clarridgeae) (3, 7, 9, 16, 17, 21, 40, 44), bacillary angiomatosis in immunocompromised patients (B. quintana, B. henselae) (14,30,48), neuroretinitis (B. grahamii) (28), and cat scratch disease (B. henselae, B. clarridgeiae) (5,31,42).Because Bartonella species frequently induce persistent intravascular infections, it has been difficult to attribute chronic disease causation to infection in humans and companion animals; much of this difficulty may be related to the few and often very subtle clinical abnormalities that are reported by a patient or observed in a sick animal. Confirming disease causation is especially difficult in retrospective or prospective animal studies in which Bartonella bacteremia can be detected in overtly healthy, natural reservoir hosts-a paradigm in opposition to Koch's postulates for disease causation (12, 23). Nevertheless, an increasingly diverse spectrum of Bartonella-associated infections have been recognized in people and in dogs due to the development of new approaches to improving serologic and molecular diagnostic testing methods, which prove to be, in most instances, more sensitive than conventional culture methods for the isolation of Bartonella species (13,15,26,30,33,38,41,46,49). Primary isolation of Bartonella species following lysis centrifugation, or freezing of a blood sample, followed by application to a blood agar plate, is the most widely used method for the microbiological diagnosis of bartonellosis. Isolation of Bartonel...

BackgroundDuring a two year period, a 27-year-old female veterinarian experienced migraine headaches, seizures, including status epilepticus, and other neurological and neurocognitive abnormalities. Prior to and during her illness, she had been actively involved in hospital-based work treating domestic animals, primarily cats and dogs, in Grenada and Ireland and anatomical research requiring the dissection of wild animals (including lions, giraffe, rabbits, mongoose, and other animals), mostly in South Africa. The woman reported contact with fleas, ticks, lice, biting flies, mosquitoes, spiders and mites and had also been scratched or bitten by dogs, cats, birds, horses, reptiles, rabbits and rodents. Prior diagnostic testing resulted in findings that were inconclusive or within normal reference ranges and no etiological diagnosis had been obtained to explain the patient’s symptoms.MethodsPCR assays targeting Anaplasma spp. Bartonella spp. and hemotopic Mycoplasma spp. were used to test patient blood samples. PCR positive amplicons were sequenced directly and compared to GenBank sequences. In addition, Bartonella alpha Proteobacteria growth medium (BAPGM) enrichment blood culture was used to facilitate bacterial growth and Bartonella spp. serology was performed by indirect fluorescent antibody testing.ResultsAnaplasma platys, Bartonella henselae and Candidatus Mycoplasma haematoparvum DNA was amplified and sequenced from the woman’s blood, serum or blood culture samples. Her serum was variably seroreactive to several Bartonella sp. antigens. Despite symptomatic improvement, six months of doxycycline most likely failed to eliminate the B. henselae infection, whereas A. platys and Candidatus M. haematoparvum DNA was no longer amplified from post-treatment samples.ConclusionsAs is typical of many veterinary professionals, this individual had frequent exposure to arthropod vectors and near daily contact with persistently bacteremic reservoir hosts, including cats, the primary reservoir host for B. henselae, and dogs, the presumed primary reservoir host for A. platys and Candidatus Mycoplasma haematoparvum. Physicians caring for veterinarians should be aware of the occupational zoonotic risks associated with the daily activities of these animal health professionals.

We detected infection with a Bartonella species (B. henselae or B. vinsonii subsp. berkhoffii) in blood samples from six immunocompetent patients who presented with a chronic neurological or neurocognitive syndrome including seizures, ataxia, memory loss, and/or tremors. Each of these patients had substantial animal contact or recent arthropod exposure as a potential risk factor for Bartonella infection. Additional studies should be performed to clarify the potential role of Bartonella spp. as a cause of chronic neurological and neurocognitive dysfunction.Bartonella henselae causes a prototypical illness characterized by fever and regional lymphadenopathy following a cat scratch or bite (8, 9). Cat scratch disease (CSD) is usually self-limited, and antibiotic therapy has minimal impact on the clinical course (11, 34). However, a spectrum of neurological manifestations, including ischemic stroke, cerebral arteritis, transverse myelitis, radiculitis, grand mal seizures, epilepsia partialis continua, status epilepticus, coma, and fatal encephalitis, in patients with CSD have been described previously (21,34). Chronic neurological or neurocognitive syndromes associated with persistent Bartonella bacteremia are less well characterized. Neurological symptoms following a cat scratch have also been described in association with B. quintana infection, and recent evidence indicates that cats can harbor B. quintana (6,13,32,37).Although CSD is considered to be self-limiting, persistent intravascular infection of a child with B. henselae for 4 months after a cat scratch has been reported previously (2). Furthermore, we recently described chronic intravascular infection with both B. henselae and B. vinsonii subsp. berkhoffii in immunocompetent people with occupational animal contact and arthropod exposure (5). Cats are the primary reservoir hosts for B. henselae, whereas to date, B. vinsonii subsp. berkhoffii has been isolated only from dogs, coyotes, foxes, or people (9, 27). Domestic and wild canines serve as the primary environmental reservoir for B. vinsonii subsp. berkhoffii, and dogs can be involved in the transmission of B. vinsonii subsp. berkhoffii and B. henselae to people (7,9,10,27,36).In this study, we report the isolation of B. henselae or B. vinsonii subsp. berkhoffii from, or the molecular detection of these pathogens in, blood samples from six people who exhibited a spectrum of neurological and neurocognitive abnormalities. MATERIALS AND METHODSBlood and serum samples from six individuals and cerebrospinal fluid from one patient were submitted by an attending physician to the Intracellular Pathogens Research Laboratory, Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, North Carolina State University, Raleigh, for attempted isolation of a Bartonella species. We used a previously described approach incorporating preenrichment culture of blood in BartonellaAlphaproteobacteria growth medium (BAPGM) and PCR (16). Bacterial isolation, PCR amplification, and cloning were p...