RESUMEN
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.
Asunto(s)
Ehrlichia/clasificación , Ehrlichiosis/veterinaria , Enfermedades de los Caballos/epidemiología , Enfermedades de los Caballos/microbiología , Animales , Ehrlichia/genética , Caballos , Tipificación Molecular , Nicaragua/epidemiología , Filogenia , ARN Ribosómico 16S/genética , SerotipificaciónRESUMEN
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.
Asunto(s)
Bacteriemia/epidemiología , Infecciones por Bartonella/epidemiología , Bartonella/aislamiento & purificación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Artritis/microbiología , Bacteriemia/diagnóstico , Bacteriemia/microbiología , Bartonella/clasificación , Bartonella/genética , Infecciones por Bartonella/diagnóstico , Infecciones por Bartonella/microbiología , Niño , Preescolar , Estudios Transversales , Femenino , Humanos , Enfermedad de Lyme/epidemiología , Masculino , Persona de Mediana Edad , Tipificación Molecular , Serotipificación , Encuestas y Cuestionarios , Adulto JovenRESUMEN
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.
Asunto(s)
Rickettsia rickettsii/fisiología , Fiebre Maculosa de las Montañas Rocosas/transmisión , Garrapatas/microbiología , Adolescente , Animales , Proteínas de la Membrana Bacteriana Externa/genética , Doxiciclina/uso terapéutico , Humanos , Masculino , Mitocondrias/genética , North Carolina , ARN Ribosómico 16S/genética , Fiebre Maculosa de las Montañas Rocosas/diagnóstico , Fiebre Maculosa de las Montañas Rocosas/tratamiento farmacológico , Piel/patología , Garrapatas/genéticaRESUMEN
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.
Asunto(s)
Bacteriemia/diagnóstico , Bacteriemia/fisiopatología , Infecciones por Bartonella/diagnóstico , Infecciones por Bartonella/fisiopatología , Bartonella/aislamiento & purificación , Alucinaciones/diagnóstico , Adolescente , Antibacterianos/administración & dosificación , Anticuerpos Antibacterianos/sangre , Bacteriemia/tratamiento farmacológico , Bacteriemia/microbiología , Infecciones por Bartonella/tratamiento farmacológico , Infecciones por Bartonella/microbiología , ADN Bacteriano/sangre , Femenino , Humanos , Resultado del TratamientoRESUMEN
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.
Asunto(s)
Anaplasmosis , Enfermedades de los Perros , Ehrlichiosis , Anaplasma , Anaplasmosis/diagnóstico , Animales , Anticuerpos Antibacterianos , Enfermedades de los Perros/diagnóstico , Perros , Ehrlichia , Ehrlichiosis/diagnóstico , Ehrlichiosis/veterinaria , PéptidosRESUMEN
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.
Asunto(s)
Anaplasma phagocytophilum , Enfermedades de los Perros/transmisión , Ehrlichiosis/veterinaria , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , Complicaciones Infecciosas del Embarazo/veterinaria , Animales , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/microbiología , Perros , Doxiciclina/efectos adversos , Doxiciclina/uso terapéutico , Ehrlichiosis/tratamiento farmacológico , Ehrlichiosis/transmisión , Femenino , Embarazo , Complicaciones Infecciosas del Embarazo/tratamiento farmacológico , Complicaciones Infecciosas del Embarazo/microbiología , TeratógenosRESUMEN
Eighty-six dogs were selected based upon Ehrlichia (E.) canis SNAP 3Dx positive results to determine clinical relevance of annual E. canis screening. Immunofluorescence assay showed 72 (84%) of 86 dogs were seroreactive for E. canis. Polymerase chain reaction (PCR) revealed that 12 (14%) of 86 dogs had Ehrlichia deoxyribonucleic acid; seven had E. canis, four had E. ewingii, and one was coinfected with E. chaffeensis and E. ewingii. Thrombocytopenia (<164,000 platelets/microL) was found in 28 (39%) of 72 dogs. In this study, thrombocytopenia was frequently detected in healthy Ehrlichia SNAP 3Dx-positive dogs, whereas active infection was infrequently confirmed by PCR. Therefore, treatment based upon screening results alone is not recommended.
Asunto(s)
Enfermedades de los Perros/diagnóstico , Ehrlichiosis/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Técnica del Anticuerpo Fluorescente/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Animales , Diagnóstico Diferencial , Pruebas Diagnósticas de Rutina/métodos , Pruebas Diagnósticas de Rutina/normas , Pruebas Diagnósticas de Rutina/veterinaria , Enfermedades de los Perros/sangre , Perros , Ehrlichia/genética , Ehrlichia/inmunología , Ehrlichia/aislamiento & purificación , Ehrlichiosis/sangre , Ehrlichiosis/diagnóstico , Ensayo de Inmunoadsorción Enzimática/normas , Técnica del Anticuerpo Fluorescente/normas , Reacción en Cadena de la Polimerasa/normas , Trombocitopenia/sangre , Estados UnidosRESUMEN
The molecular characterization of a Bartonella vinsonii subsp. berkhoffii genotype III strain (NCSU strain 06-CO1) isolated from the blood of a military working dog diagnosed with endocarditis is reported in this study. Several genes were amplified and sequenced for comparative sequence similarity with other strains.
Asunto(s)
Bartonella/genética , ADN Bacteriano/genética , Animales , Bartonella/aislamiento & purificación , Infecciones por Bartonella/microbiología , Infecciones por Bartonella/veterinaria , Sangre/microbiología , ADN Bacteriano/química , Enfermedades de los Perros/microbiología , Perros , Endocarditis Bacteriana/microbiología , Femenino , Genes Bacterianos , Genotipo , Datos de Secuencia MolecularRESUMEN
To identify the tick-borne pathogens in dogs from Grenada, we conducted a serologic survey for Ehrlichia canis in 2004 (104 dogs) and a comprehensive serologic and molecular survey for a variety of tick-borne pathogens in 2006 (73 dogs). In 2004 and 2006, 44 and 32 dogs (42.3% and 43.8%) were seropositive for E. canis, respectively. In 2006, several tick-borne pathogens were identified by serology and PCR. DNA of E. canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, and Bartonella sp. were identified in 18 (24.7%), 14 (19.2%), 5 (7%), 5 (7%), and 1 (1.4%) dogs, respectively. Six (8.2%) dogs were seropositive for Bartonella vinsonii subsp. berkhoffii. All dogs were seronegative and PCR-negative for Rickettsia spp. Coinfection with two or three pathogens was observed in eight dogs. Partial 16S rRNA E. canis and A. platys sequences were identical to sequences in GenBank. Partial 18S rRNA gene sequences from the Grenadian H. canis were identical to each other and had one possible mismatch (ambiguous base) from H. canis detected from Spain and Brazil. Grenadian B. c. vogeli sequences were identical to B. c. vogeli from Brazil and Japan. All of the detected pathogens are transmitted, or suspected to be transmitted, by Rhipicephalus sanguineus. Results of this study indicate that dogs from Grenada are infected with multiple tick-borne pathogens; therefore, tick-borne diseases should be included as differentials for dogs exhibiting thrombocytopenia, leukopenia, fever, or lethargy. One pathogen, E. canis, is also of potential public health significance.
Asunto(s)
Babesiosis/veterinaria , Coccidiosis/veterinaria , Enfermedades de los Perros/epidemiología , Infecciones por Bacterias Gramnegativas/veterinaria , Enfermedades por Picaduras de Garrapatas/veterinaria , Animales , Anticuerpos Antibacterianos/sangre , Anticuerpos Antiprotozoarios/sangre , Babesia/aislamiento & purificación , Babesiosis/epidemiología , Coccidios/aislamiento & purificación , Coccidiosis/epidemiología , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/parasitología , Perros , Femenino , Bacterias Gramnegativas/aislamiento & purificación , Infecciones por Bacterias Gramnegativas/diagnóstico , Infecciones por Bacterias Gramnegativas/epidemiología , Grenada/epidemiología , Masculino , Polimorfismo de Nucleótido Simple , Prevalencia , ARN Ribosómico 16S/genética , ARN Ribosómico 18S/genética , Rhipicephalus sanguineus/microbiología , Rhipicephalus sanguineus/parasitología , Enfermedades por Picaduras de Garrapatas/diagnóstico , Enfermedades por Picaduras de Garrapatas/epidemiologíaRESUMEN
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.
Asunto(s)
Antígenos Bacterianos/inmunología , Infecciones por Bartonella/veterinaria , Bartonella/inmunología , Enfermedades de los Perros/diagnóstico , Animales , Infecciones por Bartonella/diagnóstico , Bartonella henselae/inmunología , Bartonella quintana/inmunología , Células Cultivadas , Enfermedades de los Perros/microbiología , Perros , Técnica del Anticuerpo Fluorescente/métodos , Técnica del Anticuerpo Fluorescente/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Sensibilidad y Especificidad , Pruebas Serológicas/métodos , Pruebas Serológicas/veterinaria , Fiebre de las Trincheras/diagnóstico , Fiebre de las Trincheras/veterinariaRESUMEN
A carrier state develops in some Ehrlichia canis-infected dogs due to ineffective host defenses. The subsequent development of immune-mediated diseases or opportunistic infections in chronic ehrlichiosis suggests dysregulation of immunity; however, the immunobiology of this infection has not been well characterized. In this study, eight dogs were infected with E. canis, and changes in seroreactivity, serum immunoglobulin (Ig) concentrations, peripheral blood T cell subsets, lymphocyte blastogenesis (LBT), and lymphokine-activated killer (LAK) activity were evaluated over 4 months. Infection, which was documented by seroconversion, polymerase chain reaction, and blood culture, caused self-limiting fever and thrombocytopenia. Infected dogs developed an anti-E. canis antibody response but were not immune to re-infection. Serum IgM, IgG, and IgA concentrations were unaffected by E. canis. The percentage of circulating CD4(+) T cells was similar in uninfected and infected dogs at all points. Infected dogs developed a CD8(+) lymphocytosis 6 weeks after inoculation that subsequently subsided, despite organism persistence. Functional defects of cell-mediated immunity, measured as suppression of LAK activity or mitogen-driven LBT, were not observed. These results suggest that immune responses are not grossly impaired in young dogs during the first several months following experimental E. canis infection.
Asunto(s)
Enfermedades de los Perros/inmunología , Enfermedades de los Perros/microbiología , Ehrlichia canis/inmunología , Ehrlichiosis/inmunología , Ehrlichiosis/veterinaria , Animales , Formación de Anticuerpos/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/microbiología , Perros , Doxiciclina/uso terapéutico , Ehrlichia canis/genética , Ehrlichiosis/sangre , Ehrlichiosis/microbiología , Femenino , Citometría de Flujo/veterinaria , Inmunidad Celular/inmunología , Inmunodifusión/veterinaria , Inmunoglobulinas/sangre , Inmunofenotipificación/veterinaria , Células Asesinas Activadas por Linfocinas/inmunología , Distribución Aleatoria , Organismos Libres de Patógenos EspecíficosRESUMEN
OBJECTIVE: To examine the correlation between results for an indirect immunofluorescence assay (IFA) that uses Ehrlichia canis antigen as a substrate (ie, E canis-IFA), 2 western blot (WB) analyses, and a commercially available ELISA in the detection of E canis antibody in dog sera. SAMPLE POPULATION: 54 canine serum samples that were reactive on E canis-IFA and 16 canine serum samples that were E canis-IFA nonreactive. PROCEDURE: Serum samples were evaluated by use of 2 WB analyses and a commercially available ELISA. Correlation between results of the 3 testing modalities (ie, IFA, WB analyses, and the ELISA) was examined by use of nonreactive (E canis-IFA reciprocal titer, < 20), low-titer (reciprocal titer, 80 to 160), medium-titer (reciprocal titer, 320 to 2,560), and high-titer (reciprocal titer, 5,120 to > 20,480) serum samples. RESULTS: For all serum samples in the nonreactive (n = 16), medium-titer (17), and high-titer (18) groups, correlation of results among IFA, WB analyses, and the commercially available ELISA was excellent. A poor correlation was found between IFA results and those of WB analyses and the ELISA for serum samples in the low-titer group (19), with only 4 of the 19 serum samples having positive results on both WB analyses and the commercially available ELISA. CONCLUSIONS AND CLINICAL RELEVANCE: The discrepancy between E canis-IFA, WB analyses, and the commercially available ELISA results for the low-titer serum samples may be related to a high IFA sensitivity or, more likely, a lack of specificity associated with cross-reactivity among Ehrlichia spp.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Western Blotting/veterinaria , Enfermedades de los Perros/diagnóstico , Ehrlichia canis/inmunología , Ehrlichiosis/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Animales , Enfermedades de los Perros/sangre , Enfermedades de los Perros/inmunología , Perros , Ehrlichia canis/aislamiento & purificación , Ehrlichiosis/sangre , Ehrlichiosis/diagnóstico , Ehrlichiosis/inmunologíaRESUMEN
Infectious diseases caused by pathogens transmitted by ticks and other insect vectors are an important cause of morbidity and mortality in both dogs and humans throughout North America. The purpose of this study was to determine the seroprevalence of selected vector-transmitted pathogens in southern Ontario and Quebec. Samples submitted to the Vector Borne Disease Diagnostic Laboratory (VBDDL) at the North Carolina State University College of Veterinary Medicine were evaluated for antibodies to Ehrlichia canis, Anaplasma phagocytophilum, Babesia canis, Bartonella henselae, Borrelia burgdorferi, Bartonella vinsonii subspecies berkhoffii, and Rickettsia rickettsii. Information regarding breed and the city or province from which the sample originated was recorded; however, travel history was unknown for the majority of dogs. Overall seroprevalence to these tick-borne pathogens in southern Ontario and Quebec is low compared with most regions of the United States, suggesting that veterinarians in this region of Canada should pursue diagnostic evidence of infection in dogs with a travel history or prior residence in areas endemic for exposure to tick-borne infections.
Asunto(s)
Vectores Arácnidos , Enfermedades de los Perros/epidemiología , Enfermedades por Picaduras de Garrapatas/veterinaria , Garrapatas/microbiología , Garrapatas/parasitología , Animales , Anticuerpos Antibacterianos/sangre , Anticuerpos Antiprotozoarios/sangre , Vectores Arácnidos/microbiología , Vectores Arácnidos/parasitología , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/parasitología , Perros , Femenino , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Masculino , Ontario/epidemiología , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/veterinaria , Quebec/epidemiología , Estudios Seroepidemiológicos , Enfermedades por Picaduras de Garrapatas/epidemiología , Enfermedades por Picaduras de Garrapatas/microbiología , Enfermedades por Picaduras de Garrapatas/parasitología , ViajeRESUMEN
Ehrlichia species are the etiological agents of emerging and life-threatening tick-borne human zoonoses, in addition to causing serious and fatal infections in companion animals and livestock. We developed the first tricolor TaqMan real-time polymerase chain reaction assay capable of simultaneously detecting and discriminating medically important ehrlichiae in a single reaction. Analytical sensitivity of 50 copies per reaction was attained with templates from Ehrlichia chaffeensis, Ehrlichia ewingii, and Ehrlichia canis by amplifying the genus-specific disulfide bond formation protein gene (dsb). Ehrlichia genus-specific dsb primers amplified DNA from all known Ehrlichia species but not from other rickettsial organisms including Anaplasma platys, Anaplasma phagocytophilum, Rickettsia conorii, or Rickettsia typhi. High species specificity was attained as each species-specific TaqMan probe (E. chaffeensis, E. ewingii, and E. canis) identified homologous templates but did not cross-hybridize with heterologous Ehrlichia templates at concentrations as high as 10(8) copies. Identification of E. chaffeensis, E. ewingii, and E. canis from natural and experimental infections, previously confirmed by polymerase chain reaction and serological or microscopic evidence, demonstrated the comparable specificity and sensitivity of the dsb real-time assay. This assay provides a powerful tool for prospective medical diagnosis for human and canine ehrlichioses and for ecologic and epidemiological studies involving arthropod and mammalian hosts.
Asunto(s)
Ehrlichia/genética , Ehrlichia/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Proteína Disulfuro Reductasa (Glutatión)/genética , Polimerasa Taq/metabolismo , Animales , Color , Perros , Ehrlichia/clasificación , Ehrlichia/enzimología , Ehrlichiosis/sangre , Ehrlichiosis/microbiología , Humanos , Proteína Disulfuro Reductasa (Glutatión)/metabolismo , ARN Ribosómico 16S/genética , Sensibilidad y Especificidad , Factores de TiempoRESUMEN
A 7-year-old Labrador retriever had nonspecific clinical signs that included lethargy, malaise, and difficult ambulation. The dog was native to Vancouver Island, British Columbia, and had never left this area. Morulae were identified in polymorphonuclear cells. Serologic studies and polymerase chain reaction (PCR) testing confirmed canine anaplasmosis caused by Anaplasma phagocytophilum. The dog recovered after treatment with tetracycline.
Asunto(s)
Anaplasma phagocytophilum/aislamiento & purificación , Antibacterianos/uso terapéutico , Enfermedades de los Perros/diagnóstico , Ehrlichiosis/veterinaria , Tetraciclina/uso terapéutico , Anaplasma phagocytophilum/inmunología , Animales , Colombia Británica , Enfermedades de los Perros/sangre , Enfermedades de los Perros/tratamiento farmacológico , Perros , Ehrlichiosis/sangre , Ehrlichiosis/diagnóstico , Ehrlichiosis/tratamiento farmacológico , Masculino , Resultado del TratamientoRESUMEN
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.
Asunto(s)
Anaplasma/aislamiento & purificación , Anaplasmosis/diagnóstico , Anticuerpos Antibacterianos/sangre , Enfermedades de los Gatos/diagnóstico , Ehrlichia/aislamiento & purificación , Ehrlichiosis/veterinaria , Anaplasma/genética , Anaplasma/inmunología , Anaplasmosis/sangre , Anaplasmosis/microbiología , Animales , Enfermedades de los Gatos/sangre , Enfermedades de los Gatos/microbiología , Gatos , Ehrlichia/genética , Ehrlichia/inmunología , Ehrlichiosis/sangre , Ehrlichiosis/diagnóstico , Ehrlichiosis/microbiología , Femenino , Masculino , Péptidos/genética , Reacción en Cadena de la Polimerasa , Estudios Prospectivos , Especificidad de la EspecieRESUMEN
Bartonella vinsonii subspecies berkhoffii is a recently recognized zoonotic pathogen that causes endocarditis, granulomatous rhinitis, and granulomatous lymphadenitis in dogs. Isolation of B. vinsonii (berkhoffii) from blood or tissue samples is frequently unsuccessful; therefore, diagnosis is primarily dependent on serologic or molecular testing modalities. Because previous canine serologic studies have used an indirect immunofluorescence assay (IFA), without Western immunoblot (WI) confirmation, the overall objective of this study was to examine the diagnostic use of WI for confirmation of B. vinsonii (berkhoffii) infection in dogs. To confirm that agar-grown and cell culture-grown organisms yielded similar patterns of WI antigenic protein recognition, the 2 preparations were compared using IFA-reactive sera obtained from dogs experimentally infected with B. vinsonii (berkhoffii). Temporal changes in the pattern of antigenic protein recognition were characterized using sera obtained from dogs at various time points after experimental B. vinsonii (berkhoffii) infection. The specificity of B. vinsonii (berkhoffii) WI was examined by testing canine sera that were reactive to B. henselae, B. clarridgeiae, Ehrlichia canis, Rickettsia rickettsii, Babesia canis, Anaplasma phagocytophilum (previously E. equi), or Brucella canis antigens. Clinical accessions including serum samples obtained from B. vinsonii (berkhoffii) culture-positive dogs and B. vinsonii (berkhoffii) culture-negative dogs that were IFA seroreactive to B. vinsonii (berkhoffii) antigens were examined by WI. The results of this study indicate that WI using agar-grown or cell culture-grown B. vinsonii (berkhoffii) antigens produce identical patterns of antigenic protein recognition. After experimental infection, there is a progressive increase in the number of antigenic proteins that are recognized by WI, with the 33-kD antigen representing the first and the most persistent antigen recognized by B. vinsonii (berkhoffii)-infected dogs. Regarding specificity, sera from dogs that were reactive to various heterologous antigens did not recognize B. vinsonii (berkhoffii) antigens by IFA or WI, and sera from dogs experimentally infected with B. henselae did not recognize B. vinsonii (berkhoffii) antigens by WI. Regarding clinical accessions, there was good agreement between B. vinsonii (berkhoffii) IFA test results and WI analysis. Western immunoblot analysis can be used to detect or confirm exposure to B. vinsonii (berkhoffii) in dogs.
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Antígenos Virales/análisis , Infecciones por Bartonella/veterinaria , Bartonella/patogenicidad , Western Blotting/veterinaria , Enfermedades de los Perros/microbiología , Animales , Bartonella/inmunología , Infecciones por Bartonella/inmunología , Diagnóstico Diferencial , Enfermedades de los Perros/inmunología , Perros , Sensibilidad y EspecificidadRESUMEN
Although endemic throughout much of the world, autochthonous visceral leishmaniasis has been reported on only 3 previous occasions in North America. After diagnosis of visceral leishmaniasis in 4 foxhounds from a kennel in Dutchess County, New York (index kennel), serum and ethylenediamine-tetraacetic acid (EDTA)-anticoagulated blood were collected from the remaining 108 American or cross-bred foxhounds in the index kennel and from 30 Beagles and Basset Hounds that were periodically housed in the index kennel. Samples were analyzed for antibodies to or DNA of tickborne disease pathogens and Leishmania spp. Most dogs had antibodies to Rickettsia spp., Ehrlichia spp., Babesia spp., or some combination of these pathogens but not to Bartonella vinsonii (berkhoffi). However, DNA of rickettsial, ehrlichial, or babesial agents was detected in only 9 dogs. Visceral leishmaniasis was diagnosed in 46 of 112 (41%) foxhounds from the index kennel but was not diagnosed in any of the Beagles and Basset Hounds. A positive Leishmania status was defined by 1 or more of the following criteria: a Leishmania antibody titer > or = 1:64, positive Leishmania polymerase chain reaction (PCR), positive Leishmania culture, or identification of Leishmania amastigotes by cytology or histopathology. The species and zymodeme of Leishmania that infected the foxhounds was determined to be Leishmania infantum MON-1 by isoenzyme electrophoresis. Foxhounds that were > 18 months of age or that had traveled to the southeastern United States were more likely to be diagnosed with visceral leishmaniasis. Transmission of Leishmania spp. in kennel outbreaks may involve exposure to an insect vector, direct transmission, or vertical transmission.
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Anticuerpos Antiprotozoarios/sangre , Brotes de Enfermedades/veterinaria , Enfermedades de los Perros/epidemiología , Leishmania infantum/inmunología , Leishmaniasis Visceral/veterinaria , Animales , Babesia/inmunología , Babesia/aislamiento & purificación , Bartonella/inmunología , Bartonella/aislamiento & purificación , Cartilla de ADN , ADN Protozoario/sangre , Enfermedades de los Perros/etiología , Perros , Ehrlichia/inmunología , Ehrlichia/aislamiento & purificación , Técnica del Anticuerpo Fluorescente/veterinaria , Vivienda para Animales , Leishmania infantum/clasificación , Leishmania infantum/genética , Leishmania infantum/aislamiento & purificación , Leishmaniasis Visceral/epidemiología , New York/epidemiología , Reacción en Cadena de la Polimerasa/veterinaria , Rickettsia/inmunología , Rickettsia/aislamiento & purificaciónRESUMEN
Currently, the pathogenic role of Ehrlichia canis in cats has been proposed predominantly on the basis of the serologic evidence of natural infection and the infrequent detection of morulae-like structures within the cytoplasm of leukocytes in cats. The purpose of this report was to provide molecular evidence supporting E. canis-like infection in 3 cats that had clinical manifestations consistent with canine ehrlichiosis but lacked antibodies to E. canis antigens. Serum from all 3 cats contained antinuclear antibodies (ANAs). The predominant disease manifestation was polyarthritis in 1 cat and bone marrow hypoplasia or dysplasia. accompanied by pancytopenia or anemia and thrombocytopenia, in 1 cat each. The alignment of E. canis partial 16S ribosomal DNA (rDNA: 382 nucleotide positions), amplified from EDTA blood samples from each cat, was identical to each other and was identical to a canine isolate of E. canis (GenBank accession number AF373613). In 1 cat, concurrent treatment with corticosteroids may have interfered with the therapeutic effectiveness of doxycycline for the elimination of E. canis-like infection. To further define the spectrum of ehrlichiosis in cats, polymerase chain reaction (PCR) testing may be necessary until serologic testing is thoroughly validated in experimentally or naturally infected cats. In addition, until E. canis has been isolated from cats and several tissue culture isolates are available from disparate geographic regions for detailed comparative genetic study, the molecular evidence presented in this study supporting E. canis-like infection in cats must be interpreted with caution.
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Enfermedades de los Gatos/microbiología , Ehrlichia canis/patogenicidad , Ehrlichiosis/veterinaria , Animales , Artritis/etiología , Artritis/veterinaria , Secuencia de Bases , Gatos , Cartilla de ADN , ADN Bacteriano/análisis , Diagnóstico Diferencial , Ehrlichia canis/genética , Ehrlichia canis/inmunología , Ehrlichia canis/aislamiento & purificación , Ehrlichiosis/diagnóstico , Femenino , Geografía , Inmunoglobulina G/análisis , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/veterinaria , Pruebas Serológicas , Trombocitopenia/etiología , Trombocitopenia/veterinariaRESUMEN
OBJECTIVE: To determine the prevalence of stray dogs in eastern Tennessee seropositive to Ehrlichia canis and examine the correlation between results for an ELISA, indirect immunofluorescent antibody (IFA) test, and polymerase chain reaction (PCR) assay. SAMPLE POPULATION: Blood samples obtained from 90 adult dogs admitted to an animal shelter in eastern Tennessee. PROCEDURE: Serum samples were analyzed for antibodies against E. canis by use of a commercially available ELISA kit, 2 IFA tests, and a PCR assay; testing was performed at the University of Tennessee (TN) and North Carolina State University (NCSU). The PCR amplification was performed by use of DNA extracted from EDTA-anticoagulated blood and primers designed to amplify DNA of Ehrlichia spp. RESULTS: Antibodies against E. canis were detected in only 1 dog by use of the ELISA. By IFA testing at TN, 10 of 90 (11%) dogs were seroreactive against E. canis antigens, all of which had medium to high titers to E. canis. Only 5 of the 10 TN seroreactors were also reactive against E. canis antigens in IFA tests conducted at NCSU, and all 5 had low to medium titers. The DNA of Ehrlichia spp was not amplified in any blood samples by use of PCR assays conducted at the TN or NCSU. CONCLUSIONS AND CLINICAL RELEVANCE: The discordant ELISA, IFA, and PCR results obtained in this study were unexpected and may have been related to exposure of dogs to an Ehrlichia species other than E. canis, such as E. ewingii.