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1.
Pathogens ; 10(7)2021 Jun 23.
Article in English | MEDLINE | ID: mdl-34201572

ABSTRACT

Currently, a gold standard diagnostic test for Bartonella infection in dogs is lacking. This represents a critical limitation for the development and evaluation of new diagnostic tests, as well as for the diagnosis of, and research on, bartonellosis in dogs. This retrospective observational study aims to compare the results of commonly performed and newly-reported Bartonella spp. diagnostic tests in banked clinical specimens from 90 dogs with hemangiosarcoma (HSA) using composite reference standard (CRS) and random effects latent class analysis (RE-LCA) techniques. Samples from each dog were tested using six serological or molecular diagnostic assays, including indirect fluorescent antibody (IFA) and Western blot (WB) for the detection of antibodies in serum, and qPCR and droplet digital PCR (ddPCR) in blood and fresh frozen tissue biopsy samples (mainly splenic HSA tumors and histopathologically normal spleen or skin/adipose tissue). Bartonella infection prevalence was estimated to be 78% based on the CRS (parallel testing with all six assays), and 64% based on the RE-LCA model. The assay with the highest diagnostic accuracy was qPCR performed on fresh frozen tissue biopsy samples (sensitivity: 94% by RE-LCA and 80% by CRS; specificity: 100%). When comparing newly-reported to traditional Bartonella diagnostic assays, ddPCR was more sensitive for the detection of Bartonella DNA than qPCR when testing blood samples (36% vs. 0%, p < 0.0001). Dogs that were positive on serological assays alone with negative molecular assays were highly unlikely (<3%) to be classified as infected by the RE-LCA model. These data indicate that Bartonella spp. DNA can be PCR amplified from fresh frozen tissues from a majority of dogs with HSA using both qPCR and ddPCR, supporting the use of these methods for future controlled studies comparing the prevalence of Bartonella spp. DNA in the tissue of dogs with HSA to that of unaffected controls.

2.
Pathogens ; 10(3)2021 Mar 10.
Article in English | MEDLINE | ID: mdl-33802018

ABSTRACT

Bartonella bacilliformis (B. bacilliformis), Bartonella henselae (B. henselae), and Bartonella quintana (B. quintana) are bacteria known to cause verruga peruana or bacillary angiomatosis, vascular endothelial growth factor (VEGF)-dependent cutaneous lesions in humans. Given the bacteria's association with the dermal niche and clinical suspicion of occult infection by a dermatologist, we determined if patients with melanoma had evidence of Bartonella spp. infection. Within a one-month period, eight patients previously diagnosed with melanoma volunteered to be tested for evidence of Bartonella spp. exposure/infection. Subsequently, confocal immunohistochemistry and PCR for Bartonella spp. were used to study melanoma tissues from two patients. Blood from seven of the eight patients was either seroreactive, PCR positive, or positive by both modalities for Bartonella spp. exposure. Subsequently, Bartonella organisms that co-localized with VEGFC immunoreactivity were visualized using multi-immunostaining confocal microscopy of thick skin sections from two patients. Using a co-culture model, B. henselae was observed to enter melanoma cell cytoplasm and resulted in increased vascular endothelial growth factor C (VEGFC) and interleukin 8 (IL-8) production. Findings from this small number of patients support the need for future investigations to determine the extent to which Bartonella spp. are a component of the melanoma pathobiome.

3.
Pathogens ; 9(12)2020 Dec 04.
Article in English | MEDLINE | ID: mdl-33291688

ABSTRACT

Bartonella species are globally important emerging pathogens that were not known to infect animals or humans in North America prior to the human immunodeficiency virus (HIV) epidemic. Ongoing improvements in diagnostic testing modalities have allowed for the discovery of Bartonella species (spp.) DNA in blood; cerebrospinal fluid; and the skin of patients with cutaneous lesions, fatigue, myalgia, and neurological symptoms. We describe Bartonella spp. test results for participants reporting neuropsychiatric symptoms, the majority of whom reported the concurrent development of cutaneous lesions. Study participants completed a medical history, a risk factor questionnaire, and provided cutaneous lesion photographs. Bartonella spp. serology and Bartonella alpha proteobacteria enrichment blood culture/PCR were assessed. Within a 14-month period, 33 participants enrolled; 29/33 had serological and/or PCR evidence supporting Bartonella spp. infection, of whom 24 reported concurrent cutaneous lesions since neuropsychiatric symptom onset. We conclude that cutaneous lesions were common among people reporting neuropsychiatric symptoms and Bartonella spp. infection or exposure. Additional studies, using sensitive microbiological and imaging techniques, are needed to determine if, or to what extent, Bartonella spp. might contribute to cutaneous lesions and neuropsychiatric symptoms in patients.

4.
PLoS One ; 15(1): e0227234, 2020.
Article in English | MEDLINE | ID: mdl-31923195

ABSTRACT

Hemangiosarcoma (HSA), a locally invasive and highly metastatic endothelial cell neoplasm, accounts for two-thirds of all cardiac and splenic neoplasms in dogs. Bartonella spp. infection has been reported in association with neoplastic and non-neoplastic vasoproliferative lesions in animals and humans. The objective of this study was to determine the prevalence of Bartonella spp. in conjunction with two other hemotropic pathogens, Babesia spp. and hemotropic Mycoplasma spp., in tissues and blood samples from 110 dogs with histopathologically diagnosed HSA from throughout the United States. This was a retrospective, observational study using clinical specimens from 110 dogs with HSA banked by the biospecimen repository of the Canine Comparative Oncology and Genomics Consortium. Samples provided for this study from each dog included: fresh frozen HSA tumor tissue (available from n = 100 of the 110 dogs), fresh frozen non-tumor tissue (n = 104), and whole blood and serum samples (n = 108 and 107 respectively). Blood and tissues were tested by qPCR for Bartonella, hemotropic Mycoplasma, and Babesia spp. DNA; serum was tested for Bartonella spp. antibodies. Bartonella spp. DNA was amplified and sequenced from 73% of dogs with HSA (80/110). In contrast, hemotropic Mycoplasma spp. DNA was amplified from a significantly smaller proportion (5%, p<0.0001) and Babesia spp. DNA was not amplified from any dog. Of the 100 HSA tumor samples submitted, 34% were Bartonella PCR positive (32% of splenic tumors, 57% of cardiac tumors, and 17% of other tumor locations). Of 104 non-tumor tissues, 63% were Bartonella PCR positive (56% of spleen samples, 93% of cardiac samples, and 63% of skin/subcutaneous samples). Of dogs with Bartonella positive HSA tumor, 76% were also positive in non-tumor tissue. Bartonella spp. DNA was not PCR amplified from whole blood. This study documented a high prevalence of Bartonella spp. DNA in dogs with HSA from geographically diverse regions of the United States. While 73% of all tissue samples from these dogs were PCR positive for Bartonella DNA, none of the blood samples were, indicating that whole blood samples do not reflect tissue presence of this pathogen. Future studies are needed to further investigate the role of Bartonella spp. in the development of HSA.


Subject(s)
Babesia/genetics , Babesiosis/epidemiology , Bartonella Infections/epidemiology , Bartonella Infections/veterinary , Bartonella/genetics , Dog Diseases/epidemiology , Hemangiosarcoma/epidemiology , Hemangiosarcoma/veterinary , Mycoplasma Infections/epidemiology , Mycoplasma Infections/veterinary , Mycoplasma/genetics , Animals , Babesiosis/parasitology , Bartonella Infections/microbiology , DNA, Bacterial/genetics , Dog Diseases/microbiology , Dog Diseases/parasitology , Dogs , Female , Hemangiosarcoma/microbiology , Hemangiosarcoma/parasitology , Male , Polymerase Chain Reaction , Prevalence , Retrospective Studies , United States/epidemiology
5.
J Vet Intern Med ; 34(1): 145-159, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31891215

ABSTRACT

BACKGROUND: Dynamics of infection by Bartonella and Rickettsia species, which are epidemiologically associated in dogs, have not been explored in a controlled setting. OBJECTIVES: Describe an outbreak investigation of occult Bartonella spp. infection among a group of dogs, discovered after experimentally induced Rickettsia rickettsii (Rr) infection. ANIMALS: Six apparently healthy purpose-bred Beagles obtained from a commercial vendor. METHODS: Retrospective and prospective study. Dogs were serially tested for Bartonella spp. and Rr using serology, culture, and PCR, over 3 study phases: 3 months before inoculation with Rr (retrospective), 6 weeks after inoculation with Rr (retrospective), and 8 months of follow-up (prospective). RESULTS: Before Rr infection, 1 dog was Bartonella henselae (Bh) immunofluorescent antibody assay (IFA) seroreactive and 1 was Rickettsia spp. IFA seroreactive. After inoculation with Rr, all dogs developed mild Rocky Mountain spotted fever compatible with low-dose Rr infection, seroconverted to Rickettsia spp. within 4-11 days, and recovered within 1 week. When 1 dog developed ear tip vasculitis with intra-lesional Bh, an investigation of Bartonella spp. infection was undertaken. All dogs had seroconverted to 1-3 Bartonella spp. between 7 and 18 days after Rr inoculation. Between 4 and 8 months after Rr inoculation, Bh DNA was amplified from multiple tissues from 2 dogs, and Bartonella vinsonii subsp. berkhoffii (Bvb) DNA was amplified from 4 of 5 dogs' oral swabs. CONCLUSIONS AND CLINICAL IMPORTANCE: Vector-borne disease exposure was demonstrated in research dogs from a commercial vendor. Despite limitations, our results support the possibilities of recrudescence of chronic subclinical Bartonella spp. infection after Rr infection and horizontal direct-contact transmission between dogs.


Subject(s)
Bartonella Infections/veterinary , Bartonella/isolation & purification , Dog Diseases/microbiology , Rocky Mountain Spotted Fever/veterinary , Animals , Bartonella Infections/microbiology , Coinfection , Dog Diseases/transmission , Dogs , Female , Housing, Animal , Laboratory Animal Science , Rickettsia rickettsii , Rocky Mountain Spotted Fever/complications , Rocky Mountain Spotted Fever/microbiology , Serologic Tests
6.
J Cent Nerv Syst Dis ; 11: 1179573519832014, 2019.
Article in English | MEDLINE | ID: mdl-30911227

ABSTRACT

BACKGROUND: With the advent of more sensitive culture and molecular diagnostic testing modalities, Bartonella spp. infections have been documented in blood and/or cerebrospinal fluid specimens from patients with diverse neurological symptoms. Pediatric acute-onset neuropsychiatric syndrome (PANS) is characterized by an unusually abrupt onset of cognitive, behavioral, or neurological symptoms. Between October 2015 and January 2017, a 14-year-old boy underwent evaluation by multiple specialists for sudden-onset psychotic behavior (hallucinations, delusions, suicidal and homicidal ideation). METHODS: In March 2017, Bartonella spp. serology (indirect fluorescent antibody assays) and polymerase chain reaction (PCR) amplification, DNA sequencing, and Bartonella enrichment blood culture were used on a research basis to assess Bartonella spp. exposure and bloodstream infection, respectively. PCR assays targeting other vector-borne infections were performed to assess potential co-infections. RESULTS: For 18 months, the boy remained psychotic despite 4 hospitalizations, therapeutic trials involving multiple psychiatric medication combinations, and immunosuppressive treatment for autoimmune encephalitis. Neurobartonellosis was diagnosed after cutaneous lesions developed. Subsequently, despite nearly 2 consecutive months of doxycycline administration, Bartonella henselae DNA was PCR amplified and sequenced from the patient's blood, and from Bartonella alphaproteobacteria growth medium enrichment blood cultures. B henselae serology was negative. During treatment with combination antimicrobial chemotherapy, he experienced a gradual progressive decrease in neuropsychiatric symptoms, cessation of psychiatric drugs, resolution of Bartonella-associated cutaneous lesions, and a return to all pre-illness activities. CONCLUSIONS: This case report suggests that B henselae bloodstream infection may contribute to progressive, recalcitrant neuropsychiatric symptoms consistent with PANS in a subset of patients.

7.
Vector Borne Zoonotic Dis ; 19(4): 234-241, 2019 04.
Article in English | MEDLINE | ID: mdl-30589638

ABSTRACT

Historically, Bartonella spp. have been associated with febrile illness (Oroya fever, trench fever, and cat scratch disease), endocarditis (numerous Bartonella spp.), and vasoproliferative lesions (Bartonella bacilliformis, Bartonella quintana, Bartonella henselae, and Bartonella vinsonii subsp. berkhoffii), occurring most often but not exclusively in immunocompromised patients. Recently, bloodstream infections with various Bartonella spp. have been documented in nonimmunocompromised individuals in association with a spectrum of cardiovascular, neurologic, and rheumatologic symptoms. As documented in this family, symptoms for which the medical implications remain unclear can occur in multiple family members infected with one or more Bartonella spp. Serial serologic and molecular microbiological findings supported exposure to or infection with Bartonella spp. in all seven family members. Either antibiotics failed to eliminate bacteremic infection, resulted in partial resolution of symptoms, or potentially reinfection occurred during the 19-month study period. There is a substantial need for clinical research to clarify the extent to which Bartonella spp. bacteremia induces nonspecific cardiovascular, neurologic, or rheumatologic symptoms, for ongoing improvement in the sensitivity and specificity of diagnostic testing, and clarification as to if, when, and how to treat patients with documented Bartonella spp. bacteremia.


Subject(s)
Bacteremia/microbiology , Bartonella Infections/microbiology , Bartonella/isolation & purification , Adult , Bartonella/genetics , Canada , Child , Family , Female , Humans , Male , Real-Time Polymerase Chain Reaction
8.
Vet Dermatol ; 29(6): 537-e180, 2018 Dec.
Article in English | MEDLINE | ID: mdl-30318847

ABSTRACT

BACKGROUND: Bartonella henselae, a Gram-negative, zoonotic, alpha-proteobacteria has been previously implicated in association with cutaneous vasoproliferative lesions (bacillary angiomatosis), nodular panniculitis and multifocal erythema (erythema multiforme) in dogs. OBJECTIVE: Describe clinical, microbiological and histological lesions in a dog with ear margin vasculitis and B. henselae infection. ANIMALS: A 12-month-old, specific pathogen-free intact female beagle dog maintained in a vector-free laboratory animal resource facility. METHODS AND MATERIALS: Bartonella and Rickettsia serological evaluation, Bartonella and Rickettsia PCR, Bartonella alpha-proteobacteria growth medium (BAPGM) enrichment blood culture/PCR, histopathological investigation and confocal immunohistochemical evaluation. RESULTS: Serological investigation (seroreversion) and PCR testing of aural tissue biopsies failed to support Rickettsia rickettsii as a cause of the aural vasculitis; however, B. henselae, genotype San Antonio 2 DNA was amplified and sequenced from both ear tip margins and from normal-appearing abdominal skin. Seroconversion to B. henselae was documented retrospectively by IFA testing. Bartonella henselae organisms were visualized by confocal immunostaining within all three biopsies. Histopathology revealed small vessel necrotizing vasculitis and dermal necrosis. Bartonella henselae seroreversion and complete resolution of skin lesions occurred in conjunction with administration of oral doxycycline and enrofloxacin for six weeks. CONCLUSIONS AND CLINICAL IMPORTANCE: Bartonella henselae is an emerging zoonotic pathogen that has been associated with leucocytoclastic vasculitis in humans and may have had a contributing or causative role in the development of the cutaneous aural margin vasculitis in this beagle.


Subject(s)
Bartonella henselae , Cat-Scratch Disease/veterinary , Dog Diseases/diagnosis , Ear, External/pathology , Vasculitis/veterinary , Animals , Bartonella henselae/genetics , Cat-Scratch Disease/pathology , Dog Diseases/microbiology , Dog Diseases/pathology , Dogs , Ear, External/microbiology , Female , Polymerase Chain Reaction/veterinary , Vasculitis/diagnosis , Vasculitis/pathology
9.
J Clin Microbiol ; 53(5): 1556-61, 2015 May.
Article in English | MEDLINE | ID: mdl-25740763

ABSTRACT

Bartonella species constitute emerging, vector-borne, intravascular pathogens that produce long-lasting bacteremia in reservoir-adapted (natural host or passive carrier of a microorganism) and opportunistic hosts. With the advent of more sensitive and specific diagnostic tests, there is evolving microbiological evidence supporting concurrent infection with one or more Bartonella spp. in more than one family member; however, the mode(s) of transmission to or among family members remains unclear. In this study, we provide molecular microbiological evidence of Bartonella henselae genotype San Antonio 2 (SA2) infection in four of six Danish family members, including a child who died of unknown causes at 14 months of age.


Subject(s)
Angiomatosis, Bacillary/diagnosis , Angiomatosis, Bacillary/microbiology , Bartonella henselae/classification , Bartonella henselae/isolation & purification , Family Health , Adolescent , Adult , Angiomatosis, Bacillary/transmission , Bartonella henselae/genetics , Child , Female , Genotype , Genotyping Techniques , Humans , Infant , Male , Middle Aged , Polymerase Chain Reaction , Sequence Analysis, DNA
10.
Vector Borne Zoonotic Dis ; 14(10): 703-9, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25325313

ABSTRACT

After raccoons were trapped and removed from under a house in New York, the owner and her two Papillon dogs became infested with numerous rat mites (Ornithonyssus bacoti). Two weeks later, both dogs developed pruritus, progressively severe vesicular lesions, focal areas of skin exfoliation, swelling of the vulva or prepuce, abdominal pain, and behavioral changes. Two months after the mite infestation, the owner was hospitalized because of lethargy, fatigue, uncontrollable panic attacks, depression, headaches, chills, swollen neck lymph nodes, and vesicular lesions at the mite bite sites. Due to ongoing illness, 3 months after the mite infestation, alcohol-stored mites and blood and serum from both dogs and the owner were submitted for Bartonella serology and Bartonella alpha Proteobacteria growth medium (BAPGM) enrichment blood culture/PCR. Bartonella henselae DNA was amplified and sequenced from blood or culture specimens derived from both dogs, the owner, and pooled rat mites. Following repeated treatments with doxycycline, both dogs eventually became B. henselae seronegative and blood culture negative and clinical signs resolved. In contrast, the woman was never B. henselae seroreactive, but was again PCR positive for B. henselae 20 months after the mite infestation, despite prior treatment with doxycycline. Clinicians and vector biologists should consider the possibility that rat mites may play a role in Bartonella spp. transmission.


Subject(s)
Antibodies, Bacterial/blood , Bartonella Infections/transmission , Bartonella henselae/immunology , Dog Diseases/transmission , Mite Infestations/parasitology , Mites/microbiology , Animals , Anti-Bacterial Agents/therapeutic use , Bartonella Infections/drug therapy , Bartonella Infections/microbiology , Bartonella henselae/genetics , Bartonella henselae/isolation & purification , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Dog Diseases/drug therapy , Dog Diseases/microbiology , Dogs , Doxycycline/therapeutic use , Female , Humans , Male , Polymerase Chain Reaction , Pruritus/drug therapy , Pruritus/microbiology , Pruritus/veterinary , Rats , Sequence Analysis, DNA , Zoonoses
11.
Parasit Vectors ; 7: 116, 2014 Mar 24.
Article in English | MEDLINE | ID: mdl-24655461

ABSTRACT

BACKGROUND: Canine vector borne diseases (CVBDs) comprise illnesses caused by a spectrum of pathogens that are transmitted by arthropod vectors. Some dogs have persistent infections without apparent clinical, hematological or biochemical abnormalities, whereas other dogs develop acute illnesses, persistent subclinical infections, or chronic debilitating diseases. The primary objective of this study was to screen healthy dogs for serological and molecular evidence of regionally important CVBDs. METHODS: Clinically healthy dogs (n = 118), comprising three different groups: Gp I blood donor candidates (n = 47), Gp II healthy dog volunteers (n = 50), and Gp III stray dogs (n = 21) were included in the study. Serum and ethylenediamine tetraacetic acid (EDTA) anti-coagulated blood specimens collected from each dog were tested for CVBD pathogens. RESULTS: Of the 118 dogs tested, 97 (82%) dogs had been exposed to or were infected with one or more CVBD pathogens. By IFA testing, 9% of Gp I, 42% of Gp II and 19% of Gp III dogs were seroreactive to one or more CVBD pathogens. Using the SNAP 4DX assay, Gp I dogs were seronegative for Anaplasma spp., Ehrlichia spp., and B. burgdorferi (Lyme disease) antibodies and D. immitis antigen. In Gp II, 8 dogs were Ehrlichia spp. seroreactive, 2 were infected with D. immitis and 1 was B. burgdorferi (Lyme disease) seroreactive. In Gp III, 6 dogs were infected with D. immitis and 4 were Ehrlichia spp. seroreactive. Using the BAPGM diagnostic platform, Bartonella DNA was PCR amplified and sequenced from 19% of Gp I, 20% of Gp II and 10% of Gp III dogs. Using PCR and DNA sequencing, 6% of Gps I and II and 19% of Gp III dogs were infected with other CVBD pathogens. CONCLUSION: The development and validation of specific diagnostic testing modalities has facilitated more accurate detection of CVBDs. Once identified, exposure to vectors should be limited and flea and tick prevention enforced.


Subject(s)
Bacterial Infections/veterinary , Dog Diseases/blood , Parasitic Diseases, Animal/blood , Animals , Bacterial Infections/blood , Bacterial Infections/epidemiology , Bacterial Infections/microbiology , Blood Donors , Dog Diseases/microbiology , Dog Diseases/parasitology , Dogs , Female , Insect Vectors , Male , North Carolina/epidemiology , Parasitic Diseases, Animal/epidemiology , Prevalence , Seroepidemiologic Studies , Serologic Tests
12.
Parasit Vectors ; 7: 127, 2014 Mar 26.
Article in English | MEDLINE | ID: mdl-24670154

ABSTRACT

BACKGROUND: Canine vector-borne diseases (CVBD) are caused by a diverse array of pathogens with varying biological behaviors that result in a wide spectrum of clinical presentations and laboratory abnormalities. For many reasons, the diagnosis of canine vector-borne infectious diseases can be challenging for clinicians. The aim of the present study was to compare CVBD serological and molecular testing as the two most common methodologies used for screening healthy dogs or diagnosing sick dogs in which a vector-borne disease is suspected. METHODS: We used serological (Anaplasma species, Babesia canis, Bartonella henselae, Bartonella vinsonii subspecies berkhoffii, Borrelia burgdorferi, Ehrlichia canis, and SFG Rickettsia) and molecular assays to assess for exposure to, or infection with, 10 genera of organisms that cause CVBDs (Anaplasma, Babesia, Bartonella, Borrelia, Ehrlichia, Francisella, hemotropic Mycoplasma, Neorickettsia, Rickettsia, and Dirofilaria). Paired serum and EDTA blood samples from 30 clinically healthy dogs (Group I) and from 69 sick dogs suspected of having one or more canine vector-borne diseases (Groups II-IV), were tested in parallel to establish exposure to or infection with the specific CVBDs targeted in this study. RESULTS: Among all dogs tested (Groups I-IV), the molecular prevalences for individual CVBD pathogens ranged between 23.3 and 39.1%. Similarly, pathogen-specific seroprevalences ranged from 43.3% to 59.4% among healthy and sick dogs (Groups I-IV). Among these representative sample groupings, a panel combining serological and molecular assays run in parallel resulted in a 4-58% increase in the recognition of exposure to or infection with CVBD. CONCLUSIONS: We conclude that serological and PCR assays should be used in parallel to maximize CVBD diagnosis.


Subject(s)
Bacterial Infections/veterinary , Dog Diseases/diagnosis , Parasitic Diseases, Animal/diagnosis , Serologic Tests/veterinary , Animals , Bacterial Infections/blood , Bacterial Infections/diagnosis , Bacterial Infections/microbiology , DNA, Bacterial/genetics , Dog Diseases/blood , Dogs , Parasitic Diseases, Animal/blood , Parasitic Diseases, Animal/parasitology
13.
Vet Immunol Immunopathol ; 156(1-2): 153-8, 2013 Nov 15.
Article in English | MEDLINE | ID: mdl-24120155

ABSTRACT

The lack of a suitable infection model remains an important obstacle for the pathophysiological understanding of Bartonella spp. The following pilot study was designed to determine whether cell culture-grown Bartonella henselae SA2 and Bartonella vinsonii subsp. berkhoffii genotype III would cause persistent bacteremia in dogs. Pre-inoculation screening established that two laboratory-raised Golden retrievers were naturally-infected with Bartonella koehlerae. Despite prior infection, one dog each was inoculated subcutaneously with 5 × 10(4)B. henselae (SA2 strain) or 3 × 10(4)B. vinsonii subsp. berkhoffii genotype III. Dogs were bled weekly for serological testing and culture using Bartonella alpha proteobacteria growth medium (BAPGM) diagnostic platform. Dog 1 seroconverted to B. henselae and Dog 2 seroconverted to B. vinsonii subsp. berkhoffii genotype III. Throughout the study period, Bartonella spp. DNA was neither amplified nor isolated in ante-mortem BAPGM enrichment blood cultures. B. henselae SA2 was isolated from a postmortem bone marrow from Dog 1 and B. koehlerae DNA was amplified from postmortem lung from Dog 2 following BAPGM enrichment culture. Limitations include lack of uninfected controls, a potentially suboptimal B. vinsonii subsp. berkhoffii inoculum and a relatively short duration of study. We conclude that following intradermal infection, sequestration of Bartonella spp. in tissues may limit diagnostic detection of these bacteria in dog blood samples.


Subject(s)
Bacteremia/veterinary , Bartonella Infections/immunology , Bartonella henselae/immunology , Dog Diseases/immunology , Animals , Antibodies, Bacterial/blood , Bacteremia/immunology , Bacteremia/microbiology , Bartonella Infections/microbiology , DNA, Bacterial/analysis , Dog Diseases/microbiology , Dogs
14.
Parasit Vectors ; 6: 101, 2013 Apr 15.
Article in English | MEDLINE | ID: mdl-23587194

ABSTRACT

BACKGROUND: Bartonella henselae is a zoonotic, alpha Proteobacterium, historically associated with cat scratch disease (CSD), but more recently associated with persistent bacteremia, fever of unknown origin, arthritic and neurological disorders, and bacillary angiomatosis, and peliosis hepatis in immunocompromised patients. A family from the Netherlands contacted our laboratory requesting to be included in a research study (NCSU-IRB#1960), designed to characterize Bartonella spp. bacteremia in people with extensive arthropod or animal exposure. All four family members had been exposed to tick bites in Zeeland, southwestern Netherlands. The mother and son were exhibiting symptoms including fatigue, headaches, memory loss, disorientation, peripheral neuropathic pain, striae (son only), and loss of coordination, whereas the father and daughter were healthy. METHODS: Each family member was tested for serological evidence of Bartonella exposure using B. vinsonii subsp. berkhoffii genotypes I-III, B. henselae and B. koehlerae indirect fluorescent antibody assays and for bacteremia using the BAPGM enrichment blood culture platform. RESULTS: The mother was seroreactive to multiple Bartonella spp. antigens and bacteremia was confirmed by PCR amplification of B. henselae DNA from blood, and from a BAPGM blood agar plate subculture isolate. The son was not seroreactive to any Bartonella sp. antigen, but B. henselae DNA was amplified from several blood and serum samples, from BAPGM enrichment blood culture, and from a cutaneous striae biopsy. The father and daughter were seronegative to all Bartonella spp. antigens, and negative for Bartonella DNA amplification. CONCLUSIONS: Historically, persistent B. henselae bacteremia was not thought to occur in immunocompetent humans. To our knowledge, this study provides preliminary evidence supporting the possibility of persistent B. henselae bacteremia in immunocompetent persons from Europe. Cat or flea contact was considered an unlikely source of transmission and the mother, a physician, reported that clinical symptoms developed following tick exposure. To our knowledge, this is the first time that a B. henselae organism has been visualized in and amplified from a striae lesion. As the tick bites occurred three years prior to documentation of B. henselae bacteremia, the mode of transmission could not be determined.


Subject(s)
Bacteremia/diagnosis , Bacteremia/microbiology , Bartonella Infections/diagnosis , Bartonella Infections/microbiology , Bartonella henselae/isolation & purification , Insect Bites and Stings/complications , Adolescent , Animals , Antibodies, Bacterial/blood , Bacteremia/pathology , Bartonella Infections/pathology , Female , Humans , Male , Middle Aged , Mothers , Netherlands , Nuclear Family
15.
Parasit Vectors ; 6: 98, 2013 Apr 15.
Article in English | MEDLINE | ID: mdl-23587343

ABSTRACT

BACKGROUND: Bartonella species comprise a group of zoonotic pathogens that are usually acquired by vector transmission or by animal bites or scratches. METHODS: PCR targeting the Bartonella 16S-23S intergenic spacer (ITS) region was used in conjunction with BAPGM (Bartonella alpha Proteobacteria growth medium) enrichment blood culture to determine the infection status of the family members and to amplify DNA from spiders and woodlice. Antibody titers to B. vinsonii subsp. berkhoffii (Bvb) genotypes I-III, B. henselae (Bh) and B. koehlerae (Bk) were determined using an IFA test. Management of the medical problems reported by these patients was provided by their respective physicians. RESULTS: In this investigation, immediately prior to the onset of symptoms two children in a family experienced puncture-like skin lesions after exposure to and presumptive bites from woodlouse hunter spiders. Shortly thereafter, the mother and both children developed hive-like lesions. Over the ensuing months, the youngest son was diagnosed with Guillain-Barre (GBS) syndrome followed by Chronic Inflammatory Demyelinating Polyradiculoneuropathy (CIDP). The older son developed intermittent disorientation and irritability, and the mother experienced fatigue, headaches, joint pain and memory loss. When tested approximately three years after the woodlouse hunter spider infestation, all three family members were Bartonella henselae seroreactive and B. henselae DNA was amplified and sequenced from blood, serum or Bartonella alpha-proteobacteria (BAPGM) enrichment blood cultures from the mother and oldest son. Also, B. henselae DNA was PCR amplified and sequenced from a woodlouse and from woodlouse hunter spiders collected adjacent to the family's home. CONCLUSIONS: Although it was not possible to determine whether the family's B. henselae infections were acquired by spider bites or whether the spiders and woodlice were merely accidental hosts, physicians should consider the possibility that B. henselae represents an antecedent infection for GBS, CIDP, and non-specific neurocognitive abnormalities.


Subject(s)
Angiomatosis, Bacillary/diagnosis , Bartonella henselae/isolation & purification , Cognition Disorders/diagnosis , Spider Bites/complications , Angiomatosis, Bacillary/complications , Animals , Child , Child, Preschool , Cognition Disorders/etiology , Family Health , Female , Humans , Male , Spiders/microbiology
16.
Emerg Infect Dis ; 18(5): 783-91, 2012 May.
Article in English | MEDLINE | ID: mdl-22516098

ABSTRACT

Bartonella spp. infection has been reported in association with an expanding spectrum of symptoms and lesions. Among 296 patients examined by a rheumatologist, prevalence of antibodies against Bartonella henselae, B. koehlerae, or B. vinsonii subsp. berkhoffii (185 [62%]) and Bartonella spp. bacteremia (122 [41.1%]) was high. Conditions diagnosed before referral included Lyme disease (46.6%), arthralgia/arthritis (20.6%), chronic fatigue (19.6%), and fibromyalgia (6.1%). B. henselae bacteremia was significantly associated with prior referral to a neurologist, most often for blurred vision, subcortical neurologic deficits, or numbness in the extremities, whereas B. koehlerae bacteremia was associated with examination by an infectious disease physician. This cross-sectional study cannot establish a causal link between Bartonella spp. infection and the high frequency of neurologic symptoms, myalgia, joint pain, or progressive arthropathy in this population; however, the contribution of Bartonella spp. infection, if any, to these symptoms should be systematically investigated.


Subject(s)
Bacteremia/epidemiology , Bartonella Infections/epidemiology , Bartonella/isolation & purification , Adolescent , Adult , Aged , Aged, 80 and over , Arthritis/microbiology , Bacteremia/diagnosis , Bacteremia/microbiology , Bartonella/classification , Bartonella/genetics , Bartonella Infections/diagnosis , Bartonella Infections/microbiology , Child , Child, Preschool , Cross-Sectional Studies , Female , Humans , Lyme Disease/epidemiology , Male , Middle Aged , Molecular Typing , Serotyping , Surveys and Questionnaires , Young Adult
17.
Diagn Microbiol Infect Dis ; 71(4): 430-7, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21996096

ABSTRACT

Serum and blood samples from 192 patients, who reported animal exposure (100.0%) and recent animal bites or scratches (88.0%), were screened for antibodies by indirect immunofluorescence assays and for bacteremia using the BAPGM (Bartonella alpha Proteobacteria growth medium) platform. Predominant symptoms included fatigue (79.2%), sleeplessness (64.1%), joint pain (64.1%), and muscle pain (63.0%). Bartonella spp. seroreactivity or bacteremia was documented in 49.5% (n = 95) and 23.9% (n = 46) of the patients, respectively; however, indirect immunofluorescence antibodies were not detected in 30.4% (n = 14) of bacteremic patients. Regarding components of the BAPGM platform, Bartonella DNA was amplified from 7.5% of blood (n = 21), 8.7% of serum (n = 25), and 10.3% of enrichment culture samples (n = 29). Polymerase chain reaction (PCR) on only extracted blood would not have detected Bartonella infection in 34.7% (16/46) of bacteremic patients. Serology, in conjunction with blood, serum, and BAPGM enrichment culture PCR, facilitates the diagnosis of Bartonella spp. bacteremia in immunocompetent patients.


Subject(s)
Bacteremia/diagnosis , Bacteremia/pathology , Bacteriological Techniques/methods , Bartonella Infections/diagnosis , Bartonella Infections/pathology , Bartonella/isolation & purification , Adolescent , Adult , Aged , Aged, 80 and over , Animals , Antibodies, Bacterial/blood , Bacteremia/microbiology , Bartonella Infections/microbiology , Bites and Stings/complications , Child , Female , Humans , Male , Middle Aged , Molecular Diagnostic Techniques/methods , Sensitivity and Specificity , Serologic Tests/methods , Skin/injuries , Young Adult
18.
J Clin Microbiol ; 49(9): 3415-7, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21734026

ABSTRACT

A young woman experiencing depression, anxiety, mood swings, severe headaches, muscle spasms, interphalangeal joint stiffness, decreased peripheral vision, diminished tactile sensation, and hallucinations was persistently Bartonella koehlerae seroreactive and bacteremic. Following antibiotic treatment, B. koehlerae antibodies and DNA were not detected and all symptoms resolved.


Subject(s)
Bacteremia/diagnosis , Bacteremia/physiopathology , Bartonella Infections/diagnosis , Bartonella Infections/physiopathology , Bartonella/isolation & purification , Hallucinations/diagnosis , Adolescent , Anti-Bacterial Agents/administration & dosage , Antibodies, Bacterial/blood , Bacteremia/drug therapy , Bacteremia/microbiology , Bartonella Infections/drug therapy , Bartonella Infections/microbiology , DNA, Bacterial/blood , Female , Humans , Treatment Outcome
19.
Emerg Infect Dis ; 17(5): 873-5, 2011 May.
Article in English | MEDLINE | ID: mdl-21529399

ABSTRACT

Only indirect or circumstantial evidence has been published to support transmission of Rickettsia rickettsii by Amblyomma americanum (lone star) ticks in North America. This study provides molecular evidence that A. americanum ticks can function, although most likely infrequently, as vectors of Rocky Mountain spotted fever for humans.


Subject(s)
Rickettsia rickettsii/physiology , Rocky Mountain Spotted Fever/transmission , Ticks/microbiology , Adolescent , Animals , Bacterial Outer Membrane Proteins/genetics , Doxycycline/therapeutic use , Humans , Male , Mitochondria/genetics , North Carolina , RNA, Ribosomal, 16S/genetics , Rocky Mountain Spotted Fever/diagnosis , Rocky Mountain Spotted Fever/drug therapy , Skin/pathology , Ticks/genetics
20.
Parasit Vectors ; 3: 76, 2010 Aug 24.
Article in English | MEDLINE | ID: mdl-20735840

ABSTRACT

BACKGROUND: Cats appear to be the primary reservoir host for Bartonella koehlerae, an alpha Proteobacteria that is most likely transmitted among cat populations by fleas (Ctenocephalides felis). Bartonella koehlerae has caused endocarditis in a dog and in one human patient from Israel, but other clinically relevant reports involving this bacterium are lacking. Despite publication of numerous, worldwide epidemiological studies designed to determine the prevalence of Bartonella spp. bacteremia in cats, B. koehlerae has never been isolated using conventional blood agar plates. To date, successful isolation of B. koehlerae from cats and from the one human endocarditis patient has consistently required the use of chocolate agar plates. RESULTS: In this study, Bartonella koehlerae bacteremia was documented in eight immunocompetent patients by PCR amplification and DNA sequencing, either prior to or after enrichment blood culture using Bartonella alpha Proteobacteria growth medium. Presenting symptoms most often included fatigue, insomnia, joint pain, headache, memory loss, and muscle pain. Four patients were also infected with Bartonella vinsonii subsp. berkhoffii genotype II. After molecular documentation of B. koehlerae infection in these patients, a serological test was developed and serum samples were tested retrospectively. Bartonella koehlerae antibodies were not detected (titers < 1:16) in 30 healthy human control sera, whereas five of eight patient samples had B. koehlerae antibody titers of 1:64 or greater. CONCLUSIONS: Although biased by a study population consisting of individuals with extensive arthropod and animal exposure, the results of this study suggest that B. koehlerae bacteremia is more common in immunocompetent people than has been previously suspected. Future studies should more thoroughly define modes of transmission and risk factors for acquiring infection with B. koehlerae. In addition, studies are needed to determine if B. koehlerae is a cause or cofactor in the development of arthritis, peripheral neuropathies or tachyarrhythmias in patients.

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