ABSTRACT
BACKGROUND: Little is known about the specificity of Bartonella spp. immunofluorescent antibody (IFA) assays in dogs. Bacteremia in sick dogs most often has been associated with Bartonella henselae (Bh), Bartonella vinsonii subspecies berkhoffii (Bvb), and Bartonella koehlerae (Bk). Clarification of the diagnostic utility of IFA serology when testing against these organisms is needed. OBJECTIVE: To evaluate the specificity of Bartonella IFA assays utilizing 6 cell culture-grown antigen preparations. ANIMALS: Archived sera from SPF dogs (n = 29) and from dogs experimentally infected with Bvb (n = 10) and Bh (n = 3). METHODS: Antibodies (Abs) to Bvb genotypes I, II, and III, Bh serotype I, strains H-1 and SA2, and to Bk were determined by IFA testing. RESULTS: Serum from naïve SPF dogs shown to be negative for Bartonella bacteremia did not react with any of the 6 Bartonella antigens by IFA testing. Dogs experimentally infected with Bvb genotype I developed Abs against homologous antigens, with no cross-reactivity to heterologous Bvb genotypes, Bh H-1, SA2 strains, or to Bk. Dogs experimentally infected with Bh serotype I developed Abs against Bh H-1, but not to Bh SA2 strain with no cross-reactive Abs to Bvb genotypes I-III or to Bk. CONCLUSIONS AND CLINICAL IMPORTANCE: Bartonella spp. Ab responses during acute experimental infections are species and type specific.
Subject(s)
Antibodies, Bacterial/blood , Antigens, Bacterial , Bacteremia/veterinary , Bartonella Infections/veterinary , Bartonella/isolation & purification , Dog Diseases/microbiology , Fluorescent Antibody Technique, Indirect/veterinary , Animals , Bacteremia/blood , Bacteremia/microbiology , Bartonella Infections/blood , Bartonella Infections/microbiology , Dog Diseases/blood , Dogs , Fluorescent Antibody Technique, Indirect/standards , Sensitivity and Specificity , Specific Pathogen-Free OrganismsABSTRACT
BACKGROUND: Bartonella alpha-Proteobacteria growth medium (BAPGM) enrichment culture has proven useful for documenting Bartonella species infection and has facilitated growth of other fastidious bacteria from human samples. PURPOSE: To report non-Bartonella bacterial isolates obtained from canine samples cultured using BAPGM enrichment culture. ANIMALS: Between 2004 and 2008, 695 specimens from 513 dogs were tested by the NCSU-IPRL using the BAPGM enrichment culture. Over the same period of time, blood samples from 270 dogs were cultured by the NCSU-CML using Bactec-Plus Aerobic/F media. METHODS: BAPGM isolates were characterized using Bartonella genus primers and 16S rDNA primers followed by DNA sequencing. NCSU medical records were retrospectively reviewed. Blood culture results from the NCSU-CML were compared with BAPGM blood culture results. RESULTS: Seventy-nine non-Bartonella isolates were obtained from 69/513 dogs. The most commonly isolated phylum was Proteobacteria (48.1%) with alpha-Proteobacteria being the most commonly isolated class. Staphylococcus and Sphingomonas were the most commonly isolated genera. The majority of the remaining isolates were bacteria that are rarely isolated from canine samples. Comparison of NCSU-CML and IPRL (BAPGM) blood culture isolates showed alpha-Proteobacteria were isolated more often from BAPGM. CONCLUSIONS AND CLINICAL IMPORTANCE: Use of insect cell culture enrichment medium, such as BAPGM, appears to enhance the growth of alpha-Proteobacteria, but also results in isolation of non-alpha-Proteobacteria from sick dogs. Future studies are needed to elucidate the utility of BAPGM and other "nonconventional" growth media and methods for isolation of fastidious organisms and to determine if these organisms play a causal role in disease development.
