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1.
Am J Vet Res ; 82(1): 71-80, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33369490

ABSTRACT

OBJECTIVE: To compare the performance of 5 synthetic peptide-based ELISAs with that of 3 commercially available immunofluorescent assays (IFAs) for serologic diagnosis of anaplasmosis and ehrlichiosis in dogs. SAMPLE: A convenience set of 109 serum samples obtained before and at various times after inoculation for 23 dogs that were experimentally infected with Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Ehrlichia chaffeensis, or Ehrlichia ewingii and 1 uninfected control dog in previous studies. PROCEDURES: All serum samples were assessed with 5 synthetic peptide-based ELISAs designed to detect antibodies against A phagocytophilum, A platys, E canis, E chaffeensis, and E ewingii and 3 whole organism-based IFAs designed to detect antibodies against A phagocytophilum, E canis, and E chaffeensis. The species-specific seroreactivity, cross-reactivity with the other tick-borne pathogens (TBPs), and diagnostic sensitivity and specificity were calculated for each assay and compared among assays. RESULTS: All serum samples obtained from dogs experimentally infected with a TBP yielded positive results on a serologic assay specific for that pathogen. In general, sensitivity was comparable between ELISAs and IFAs and tended to increase with duration after inoculation. Compared with the IFAs, the corresponding ELISAs were highly specific and rarely cross-reacted with antibodies against other TBPs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that peptide-based ELISAs had enhanced specificity relative to whole organism-based IFAs for detection of antibodies against Anaplasma and Ehrlichia spp, which should facilitate accurate diagnosis and may help detect dogs coinfected with multiple TBPs.


Subject(s)
Anaplasmosis , Dog Diseases , Ehrlichiosis , Anaplasma , Anaplasmosis/diagnosis , Animals , Antibodies, Bacterial , Dog Diseases/diagnosis , Dogs , Ehrlichia , Ehrlichiosis/diagnosis , Ehrlichiosis/veterinary , Peptides
2.
J Vet Intern Med ; 32(6): 1958-1964, 2018 Nov.
Article in English | MEDLINE | ID: mdl-30307643

ABSTRACT

BACKGROUND: Because of poor sensitivity and questionable specificity of immunofluorescent antibody assays (IFAs), serological diagnosis of Bartonella species infections in dogs remains challenging. Despite limitations, IFA testing is the historical "gold standard" for Bartonella serodiagnosis in animals and humans. Because most diagnostic laboratories test against only 1 or 2 Bartonella spp., testing against a broader panel of Bartonella antigens may enhance diagnostic sensitivity and specificity. OBJECTIVE: To evaluate the sensitivity and specificity of Bartonella IFA using 8 cell culture-grown Bartonella spp. isolates. ANIMALS: Archived serum samples from 34 Bartonella spp. naturally exposed, polymerase chain reaction (PCR)-positive dogs and from 26 PCR-negative and IFA-negative dogs. METHODS: Bartonella IFA sensitivity and specificity were assessed using cell culture-grown whole cell antigens derived from 3 Bartonella henselae (Bh) strains (Bh Houston 1, Bh San Antonio Type 2, Bh California 1), 3 Bartonella vinsonii subsp. berkhoffii genotypes (Bvb I, II, and III), Bartonella koehlerae (Bk), and Bartonella quintana (Bq). RESULTS: Only 62% of 34 Bartonella spp. PCR-positive dogs were seroreactive to any of the 8 Bartonella IFA antigens, indicating low IFA sensitivity. PCR-positive dogs were most often IFA seroreactive to Bq (n = 15), to Bvb II (n = 13), or to both (n = 9) antigens. Of the 26 previously IFA-negative/PCR-negative dogs, 4 (15%) were seroreactive using the expanded antigen panel. CONCLUSION AND CLINICAL IMPORTANCE: Despite IFA testing of dogs against 8 different Bartonella isolates, IFA sensitivity remained poor, and specificity was only 85%. Development of a reliable serological assay is needed to facilitate the diagnosis of Bartonella infection in dogs.


Subject(s)
Antigens, Bacterial/immunology , Bartonella Infections/veterinary , Bartonella/immunology , Dog Diseases/diagnosis , Animals , Bartonella Infections/diagnosis , Bartonella henselae/immunology , Bartonella quintana/immunology , Cells, Cultured , Dog Diseases/microbiology , Dogs , Fluorescent Antibody Technique/methods , Fluorescent Antibody Technique/veterinary , Polymerase Chain Reaction/veterinary , Sensitivity and Specificity , Serologic Tests/methods , Serologic Tests/veterinary , Trench Fever/diagnosis , Trench Fever/veterinary
3.
Parasit Vectors ; 8: 320, 2015 Jun 12.
Article in English | MEDLINE | ID: mdl-26062723

ABSTRACT

BACKGROUND: With the exception of Bartonella spp. or Cytauxzoon felis, feline vector-borne pathogens (FVBP) have been less frequently studied in North America and are generally under-appreciated as a clinical entity in cats, as compared to dogs or people. This study investigated selected FVBP seroreactivity and PCR prevalence in cats using archived samples. METHODS: Feline blood samples submitted to the Vector Borne Diseases Diagnostic Laboratory (VBDDL) at North Carolina State University College of Veterinary Medicine (NCSU-CVM) between 2008 and 2013 were tested using serological assays and PCR. An experimental SNAP® Multi-Analyte Assay (SNAP® M-A) (IDEXX Laboratories, Inc. Westbrook, Maine, USA) was used to screen all sera for antibodies to Anaplasma and Ehrlichia genus peptides and A. phagocytophilum, A. platys, B. burgdorferi, E. canis, E. chaffeensis, and E. ewingii species-specific peptides. PCR assays were used to amplify Anaplasma or Ehrlichia DNA from extracted ethylenediaminetetraacetic acid (EDTA)-anti-coagulated blood samples. Amplicons were sequenced to identify species. RESULTS: Overall, 7.8% (56/715) of cats were FVBP seroreactive and 3.2% (13/406) contained Anaplasma or Ehrlichia DNA. Serologically, B. burgdorferi (5.5%) was the most prevalent FVBP followed by A. phagocytophilum (1.8%). Ehrlichia spp. antibodies were found in 0.14% (12/715) of cats with species-specific seroreactivity to E. canis (n = 5), E. ewingii (n = 2) and E. chaffeensis (n = 1). Of seropositive cats, 16% (9/56) were exposed to more than one FVBP, all of which were exposed to B. burgdorferi and either A. phagocytophilum (n = 7) or E. ewingii (n = 2). Based upon PCR and DNA sequencing, 4, 3, 3, 2, and 1 cat were infected with A. phagocytophilum, A. platys, E. ewingii, E. chaffeensis and E. canis, respectively. CONCLUSIONS: Cats are exposed to and can be infected with vector-borne pathogens that commonly infect dogs and humans. To our knowledge, this study provides the first evidence for E. chaffeensis and E. ewingii infection in naturally-exposed cats in North America. Results from this study support the need for regional, serological and molecular FVBP prevalence studies, the need to further optimize serodiagnostic and PCR testing for cats, and the need for prospective studies to better characterize clinicopathological disease manifestations in cats infected with FVBP.


Subject(s)
Anaplasma/isolation & purification , Anaplasmosis/diagnosis , Antibodies, Bacterial/blood , Cat Diseases/diagnosis , Ehrlichia/isolation & purification , Ehrlichiosis/veterinary , Anaplasma/genetics , Anaplasma/immunology , Anaplasmosis/blood , Anaplasmosis/microbiology , Animals , Cat Diseases/blood , Cat Diseases/microbiology , Cats , Ehrlichia/genetics , Ehrlichia/immunology , Ehrlichiosis/blood , Ehrlichiosis/diagnosis , Ehrlichiosis/microbiology , Female , Male , Peptides/genetics , Polymerase Chain Reaction , Prospective Studies , Species Specificity
4.
Emerg Infect Dis ; 21(2): 335-8, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25625228

ABSTRACT

Ehrlichia sp. DNA was amplified from 4 Ehrlichia-seroreactive horses from Mérida, Nicaragua. Sequencing of 16S rDNA, sodB, and groEL genes indicated that the bacterium is most likely a novel Ehrlichia species. The tick vector and the potential for canine and human infection remain unknown.


Subject(s)
Ehrlichia/classification , Ehrlichiosis/veterinary , Horse Diseases/epidemiology , Horse Diseases/microbiology , Animals , Ehrlichia/genetics , Horses , Molecular Typing , Nicaragua/epidemiology , Phylogeny , RNA, Ribosomal, 16S/genetics , Serotyping
5.
Article in English | MEDLINE | ID: mdl-25405006

ABSTRACT

INTRODUCTION: Tick-borne pathogens cause a spectrum of disease manifestations in both dogs and humans. Recognizing regional and temporal shifts in exposure are important as tick distributions change. To better delineate regional exposure to canine tick-borne pathogens, an expanded set of species-specific peptides were used to detect Anaplasma phagocytophilum (Aph), Anaplasma platys (Apl), Ehrlichia canis (Ec), Ehrlichia chaffeensis (Ech), Ehrlichia ewingii (Eew), and Borrelia burgdorferi (Bb) antibodies in canine serum. METHODS: Archived canine serum samples (n=6,582) collected during 2008-2010 and in 2012 from the US, Canada, and the Caribbean were retrospectively screened for antibodies against Ehrlichia and Anaplasma species-specific peptides. Overall, regional and temporal seroprevalence rates were determined. RESULTS: Overall Bb and Eew were the most seroprevalent pathogens. During 2008-2010, seroprevalence rates increased overall for Aph and Ech, and regionally, Bb and Aph seroprevalence rates increased in the South. Canada had unexpectedly high seroprevalence rates for Ec and Apl. The most common co-exposures were Eew+Ech, followed by Aph+Bb and Eew+Bb. CONCLUSIONS: This study demonstrated significant shifts in canine vector-borne disease seroprevalence rates. The use of specific peptides facilitated improved geographic delineation of tick-borne pathogen distributions among dogs, which may enhance epidemiological surveillance of vector-borne pathogens shared by dogs and humans.

6.
Vector Borne Zoonotic Dis ; 14(10): 724-32, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25325316

ABSTRACT

Vector-borne disease (VBD) pathogens remain an emerging health concern for animals and humans throughout the world. Surveillance studies of ticks and humans have made substantial contributions to our knowledge of VBD epidemiology trends, but long-term VBD surveillance data of dogs in the United States is limited. This seroreactivity study assessed US temporal and regional trends and co-exposures to Anaplasma, Babesia, Bartonella, Borrelia burgdorferi, Dirofilaria immitis, Ehrlichia spp., and spotted fever group Rickettsia in dogs from 2004-2010. Dog serum samples (N=14,496) were submitted to the North Carolina State University, College of Veterinary Medicine, Vector Borne Disease Diagnostic Laboratory for vector-borne pathogens diagnostic testing using immunofluorescent antibody (IFA) and enzyme-linked immunosorbent assay (ELISA) assays. These convenience samples were retrospectively reviewed and analyzed. The largest proportion of samples originated from the South (47.6%), with the highest percent of seroreactive samples observed in the Midatlantic (43.4%), compared to other US regions. The overall seroreactivity of evaluated VBD antigens were Rickettsia rickettsia (10.4%), B. burgdorferi (5.2%), Ehrlichia spp. (4.3%), Bartonella henselae (3.8%), Anaplasma spp. (1.9%), Bartonella vinsonii subsp. berkhoffii (1.5%), Babesia canis (1.1%), and D. immitis (0.8%). Significant regional and annual seroreactivity variation was observed with B. burgdorferi, Ehrlichia, and Rickettsia exposures. Seasonal seroreactivity variation was evident with Rickettsia. Seroreactivity to more than one antigen was present in 16.5% of exposed dogs. Nationally, the most prevalent co-exposure was Rickettsia with Ehrlichia spp. (5.3%), and the highest odds of co-exposure was associated with Anaplasma spp. and B. burgdorferi (odds ratio=6.6; 95% confidence interval 5.0, 8.8). Notable annual and regional seroreactivity variation was observed with certain pathogens over 7 years of study, suggesting canine surveillance studies may have value in contributing to future VBD knowledge.


Subject(s)
Dog Diseases/epidemiology , Tick-Borne Diseases/veterinary , Anaplasma/immunology , Anaplasmosis/epidemiology , Animals , Arachnid Vectors/microbiology , Arachnid Vectors/parasitology , Babesia/immunology , Babesiosis/epidemiology , Bartonella/immunology , Bartonella Infections/epidemiology , Bartonella Infections/veterinary , Borrelia burgdorferi/immunology , Coinfection/veterinary , Dirofilaria immitis/immunology , Dirofilariasis/epidemiology , Dog Diseases/microbiology , Dog Diseases/parasitology , Dog Diseases/transmission , Dogs , Ehrlichia/immunology , Ehrlichiosis/epidemiology , Ehrlichiosis/veterinary , Lyme Disease/epidemiology , Retrospective Studies , Rickettsia/immunology , Rickettsia Infections/epidemiology , Rickettsia Infections/veterinary , Seroepidemiologic Studies , Tick-Borne Diseases/epidemiology , Tick-Borne Diseases/microbiology , Tick-Borne Diseases/parasitology , Ticks/microbiology , Ticks/parasitology , United States/epidemiology
7.
Am J Trop Med Hyg ; 91(6): 1161-5, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25266347

ABSTRACT

This article presents two case reports of Anaplasma platys detection in two women from Venezuela. Both patients were exposed to Rhipicephalus sanguineus, the presumed tick vector, and experienced chronic, nonspecific clinical signs including headaches and muscle pains. Intra-platelet inclusion bodies resembling A. platys were observed in buffy coat smears and A. platys DNA was amplified and sequenced from whole blood; however, treatment with doxycycline did not alleviate their symptoms. These cases provide further support for A. platys as a zoonotic tick-borne pathogen, most likely of low pathogenicity; nonetheless, the cause of illness in humans by A. platys is yet to be confirmed.


Subject(s)
Anaplasma/genetics , Anaplasmosis/diagnosis , Adult , Base Sequence , DNA Primers , DNA, Bacterial/genetics , Female , Humans , Middle Aged , Polymerase Chain Reaction , Venezuela
8.
Parasit Vectors ; 7: 298, 2014 Jul 01.
Article in English | MEDLINE | ID: mdl-24984562

ABSTRACT

BACKGROUND: Anaplasmosis, caused by Anaplasma phagocytophilum and Anaplasma platys, and ehrlichiosis, caused by Ehrlichia chaffeensis, Ehrlichia ewingii, the "Panola Mountain Ehrlichia" and Ehrlichia muris-like pathogens have been identified as emerging tick borne infectious diseases in dogs and human patients. Persistent intravascular infection with these bacteria is well documented in dogs, but is less well documented in human beings. METHODS: Serology and PCR targeting multiple microbial genes, followed by DNA sequencing, was used to test sequential blood samples. Tissue culture isolation was attempted in two laboratories. RESULTS: A. platys, E. chaffeensis, and E. ewingii DNA was amplified from two Anaplasma and Ehrlichia seronegative family members and their dog, all lacking typical symptoms of anaplasmosis or ehrlichiosis. Following treatment with doxycycline, the dog and mother were Anaplasma and Ehrlichia spp. PCR negative. CONCLUSIONS: Sequential PCR testing provided molecular evidence supporting intravascular persistence of A. platys and Ehrlichia spp. in two humans and their dog. Diagnosticians and clinicians should consider the potential for co-infections due to these tick borne organisms.


Subject(s)
Anaplasma/isolation & purification , Anaplasmosis/blood , Ehrlichia/isolation & purification , Ehrlichiosis/veterinary , Adolescent , Anaplasmosis/microbiology , Animals , Dog Diseases/blood , Dog Diseases/microbiology , Dogs , Ehrlichiosis/blood , Ehrlichiosis/microbiology , Female , Humans , Male , Middle Aged , Polymerase Chain Reaction , Serologic Tests
9.
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
10.
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
11.
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
12.
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
13.
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
14.
Vector Borne Zoonotic Dis ; 11(11): 1425-32, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21736485

ABSTRACT

Bartonella species comprise a genus of gram-negative, fastidious, intracellular bacteria that have been implicated in association with an increasing spectrum of disease manifestations in dogs and human patients. In this study, chronic canine and human disease, for which causation was not diagnostically defined, were reported by the breeder of a kennel of Doberman pinschers. In addition to other diagnostic tests, serology, polymerase chain reaction, and enrichment blood culture were used to assess the prevalence of Bartonella sp. infection in the dogs and their owner. From five dogs, Bartonella vinsonii subsp. berkhoffii genotype I, multiple Bartonella henselae strains, and a species most similar to Candidatus B. volans, a rodent-associated Bartonella sp., were amplified and sequenced from biopsy tissues, cerebrospinal fluid, or blood enrichment cultures. The owner was bacteremic with B. vinsonii subsp. berkhoffii genotype I, the same subsp. and genotype detected in one of her dogs. These results further emphasize the ecological complexity of Bartonella sp. transmission in nature.


Subject(s)
Bartonella Infections/microbiology , Bartonella/genetics , Dog Diseases/microbiology , Animal Husbandry , Animals , Bartonella/isolation & purification , Bartonella Infections/diagnosis , Bartonella Infections/transmission , Breeding , Databases, Nucleic Acid , Dog Diseases/diagnosis , Dog Diseases/transmission , Dogs , Ecology , Female , Genotype , Humans , Polymerase Chain Reaction , Virginia
15.
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
16.
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
17.
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.

18.
Parasit Vectors ; 3(1): 29, 2010 Apr 08.
Article in English | MEDLINE | ID: mdl-20377863

ABSTRACT

BACKGROUND: Bartonella vinsonii subsp. berkhoffii is an important, emerging, intravascular bacterial pathogen that has been recently isolated from immunocompetent patients with endocarditis, arthritis, neurological disease and vasoproliferative neoplasia. Vector transmission is suspected among dogs and wild canines, which are the primary reservoir hosts. This investigation was initiated to determine if pets and family members were infected with one or more Bartonella species. METHODS: PCR and enrichment blood culture in Bartonella alpha Proteobacteria growth medium (BAPGM) was used to determine infection status. Antibody titers to B. vinsonii subsp. berkhoffii genotypes I-III and B. henselae were determined using a previously described indirect fluorescent antibody test. Two patients were tested sequentially for over a year to assess the response to antibiotic treatment. RESULTS: Intravascular infection with B. vinsonii subsp. berkhoffii genotype II and Bartonella henselae (Houston 1 strain) were confirmed in a veterinarian and his daughter by enrichment blood culture, followed by PCR and DNA sequencing. Symptoms included progressive weight loss, muscle weakness, lack of coordination (the father) and headaches, muscle pain and insomnia (the daughter). B. vinsonii subsp. berkhoffii genotype II was also sequenced from a cerebrospinal fluid BAPGM enrichment culture and from a periodontal swab sample. After repeated courses of antibiotics, post-treatment blood cultures were negative, there was a decremental decrease in antibody titers to non-detectable levels and symptoms resolved in both patients. CONCLUSIONS: B. vinsonii subsp. berkhoffii and B. henselae are zoonotic pathogens that can be isolated from the blood of immunocompetent family members with arthralgias, fatigue and neurological symptoms. Therapeutic elimination of Bartonella spp. infections can be challenging, and follow-up testing is recommended. An increasing number of arthropod vectors, including biting flies, fleas, keds, lice, sandflies and ticks have been confirmed or are suspected as the primary mode of transmission of Bartonella species among animal populations and may also pose a risk to human beings.

19.
Vector Borne Zoonotic Dis ; 10(2): 117-23, 2010 Mar.
Article in English | MEDLINE | ID: mdl-19469667

ABSTRACT

We evaluated the serological and molecular prevalence of selected organisms in 145 dogs during late spring (May/June) of 2005 and in 88 dogs during winter (February) of 2007 from the Hopi Indian reservation. Additionally, in 2005, 442 ticks attached to dogs were collected and identified as Rhipicephalus sanguineus. Infection with or exposure to at least one organism was detected in 69% and 66% of the dogs in May/June 2005 and February 2007, respectively. Exposure to spotted fever group (SFG) rickettsiae was detected in 66.4% (2005) and 53.4% (2007) of dogs, but rickettsial DNA was not detected using polymerase chain reaction. Active Ehrlichia canis infection (by polymerase chain reaction) was identified in 36.6% (2005) and 36.3% (2007) of the dogs. E. canis infection was associated with SFG rickettsiae seroreactivity (p < 0.001). Anaplasma platys DNA was detected in 8.3% (2005) and 4.5% (2007) of the dogs. Babesia canis and Bartonella vinsonii berkhoffii seroprevalences were 6.7% and 1% in 2005, whereas in 2007 prevalences were 0% and 1.1%, respectively. No Bartonella spp., Ehrlichia chaffeensis, or Ehrlichia ewingii DNA was detected. Dogs on this Hopi Indian reservation were most frequently infected with E. canis or A. platys; however, more than half of the dogs were exposed to a SFG-Rickettsia species.


Subject(s)
Anaplasmosis/epidemiology , Dog Diseases/microbiology , Dog Diseases/parasitology , Ehrlichiosis/veterinary , Anaplasma/isolation & purification , Animals , Arizona/epidemiology , Dog Diseases/epidemiology , Dogs , Ehrlichia/isolation & purification , Ehrlichiosis/epidemiology , Female , Host-Pathogen Interactions , Male , Prevalence , Tick Infestations/epidemiology , Tick Infestations/veterinary
20.
J Am Anim Hosp Assoc ; 45(5): 232-8, 2009.
Article in English | MEDLINE | ID: mdl-19723846

ABSTRACT

Granulocytic anaplasmosis is an emerging infectious disease affecting dogs and humans in the United States and other regions of the world. Relatively few cases have been described in pregnant women, and perinatal transmission appears to occur infrequently in humans. Infection in pregnant dogs has not been reported. Diagnosis of infection during pregnancy poses therapeutic challenges, because doxycycline, the treatment of choice, is teratogenic. Also, infection during pregnancy may result in more severe disease. When infection is diagnosed after parturition, knowledge of the risk of perinatal transmission to offspring is important, because prophylactic therapy in neonates is also not without risk. In this report, we describe relatively severe clinical manifestations of Anaplasma phagocytophilum infection in a postpartum bitch and a lack of perinatal transmission to her puppies.


Subject(s)
Anaplasma phagocytophilum , Dog Diseases/transmission , Ehrlichiosis/veterinary , Infectious Disease Transmission, Vertical/veterinary , Pregnancy Complications, Infectious/veterinary , Animals , Dog Diseases/drug therapy , Dog Diseases/microbiology , Dogs , Doxycycline/adverse effects , Doxycycline/therapeutic use , Ehrlichiosis/drug therapy , Ehrlichiosis/transmission , Female , Pregnancy , Pregnancy Complications, Infectious/drug therapy , Pregnancy Complications, Infectious/microbiology , Teratogens
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