@article{breitschwerdt_suksawat_chomel_hegarty_2003, title={The Immunologic Response of Dogs to Bartonella Vinsonii Subspecies Berkhoffii Antigens: as Assessed by Western Immunoblot Analysis}, volume={15}, ISSN={1040-6387 1943-4936}, url={http://dx.doi.org/10.1177/104063870301500408}, DOI={10.1177/104063870301500408}, abstractNote={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.}, number={4}, journal={Journal of Veterinary Diagnostic Investigation}, publisher={SAGE Publications}, author={Breitschwerdt, Edward B. and Suksawat, Jiraporn and Chomel, Bruno and Hegarty, Barbara C.}, year={2003}, month={Jul}, pages={349–354} }
 @article{suksawat_pitulle_arraga-alvarado_madrigal_hancock_breitschwerdt_2001, title={Coinfection with Three Ehrlichia Species in Dogs from Thailand and Venezuela with Emphasis on Consideration of 16S Ribosomal DNA Secondary Structure}, volume={39}, ISSN={0095-1137}, url={http://dx.doi.org/10.1128/JCM.39.1.90-93.2001}, DOI={10.1128/JCM.39.1.90-93.2001}, abstractNote={ABSTRACT
          
            As part of a larger study to investigate tick-borne infections in dogs from Thailand and Venezuela, documentation of coinfection with three
            Ehrlichia
            species in two dogs, one from each country, became the focus of the present study. Although neither dog had clinical signs attributable to ehrlichiosis, both dogs were anemic and neutropenic and the Thai dog was thrombocytopenic. Genus- and species-specific PCR targeting the 16S rRNA genes indicated that both dogs were coinfected with
            Ehrlichia canis
            ,
            E. platys
            , and
            E. equi
            . To our knowledge, these results provide the first molecular documentation for the presence of
            E. equi
            in dogs from these countries. Using universal bacterial PCR primers, one nearly full-length 16S rRNA gene could be amplified from each dog. The sequences were identical to each other and almost identical to that of
            E. platys
            (
            AF156784
            ), providing the first
            E. platys
            16S ribosomal DNA (rDNA) sequences reported from these two geographically divergent countries. To determine whether these sequence differences allow differentiation between these two strains and other published 16S rDNA
            E. platys
            sequences, we performed a phylogenetic analysis of the rRNA, incorporating the consideration of secondary structure.
          }, number={1}, journal={Journal of Clinical Microbiology}, publisher={American Society for Microbiology}, author={Suksawat, J. and Pitulle, C. and Arraga-Alvarado, C. and Madrigal, K. and Hancock, S. I. and Breitschwerdt, E. B.}, year={2001}, month={Jan}, pages={90–93} }
 @article{suksawat_yu_hancock_hegarty_nilkumhang_breitschwerdt_2001, title={Serologic and molecular evidence of coinfection with multiple vector-borne pathogens in dogs from Thailand}, volume={15}, ISSN={["1939-1676"]}, DOI={10.1892/0891-6640(2001)015<0453:SAMEOC>2.3.CO;2}, abstractNote={Forty-nine dogs from Thailand were evaluated for serologic evidence of exposure or polymerase chain reaction (PCR) evidence of infection with vectorborne pathogens, includingEhrlichia sp. (Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia equi, and Ehr-lichia risticii), Bartonella vinsonii subsp. berkhoffi(Bvb), spotted fever group (SFG) rickettsiae(Rickettsia rickettsii), Typhus group (TG) rickettsiae(Rickettsia Canada, Rickettsia prowazekii, andRickettsia typhi), andBabesia sp. (Babesia canisandBabesia gibsonii). All study dogs had at least 1 of 3 entry criteria: fever, anemia, or thrombocytopenia. By immunofluorescence antibody (IFA) testing, seroreactivity was most prevalent to E chaffeensis (74%) and E canis (71%) antigens, followed by E equi (58%), Bvb (38%), E risticii (38%), R prowazekii (24%), B canis (20%), R rickettsii (12%), R Canada (4%), and B gibsonii (4%) antigens. There was 100% concordance between E canis IFA and Western blot immunoassay (WI) for 35 of 35 samples; 2 samples were IFA and WI reactive only to E equi antigens. By PCR amplification, 10 dogs were found to be infected with E canis, 5 with Ehrlichia platys, and 3 with B canis. Sequencing of PCR products was undertaken to compare Ehrlichia strains from Thailand to strains originating from the United States. Partial DNA sequence analysis confirmed infection with E canis and E platys, with identical 16S rRNA sequence alignment to E canis (U26740) and to E platys (M83801), as reported in GenBank. Partial E canis P28.1 and P28.2 amino acid sequences from Thai dogs were divergent from analogous sequences derived from North American E canis (AF082744) strains, suggesting that the Thai dogs were infected with a geographically distinct strain of E canis compared to North American strains. The results of this study indicate that dogs in Thailand have substantial exposure to vectorborne diseases and that coinfection with these pathogens may be common.}, number={5}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Suksawat, J and Yu, XJ and Hancock, SI and Hegarty, BC and Nilkumhang, P and Breitschwerdt, EB}, year={2001}, pages={453–462} }
 @article{suksawat_hegarty_breitschwerdt_2000, title={Seroprevalence of Ehrlichia canis, Ehrlichia equi, and Ehrlichia risticii in sick dogs from North Carolina and Virginia}, volume={14}, DOI={10.1111/j.1939-1676.2000.tb01499.x}, abstractNote={Ehrlichia canis, E equi, andE risticiiseroprevalence was determined by microimmunofluorescent antibody testing (IFA) in a sequential population of 1,845 sick dogs admitted during a 1‐year period to the North Carolina State University Veterinary Teaching Hospital. A seroreactor was defined by a reciprocal IFA titer of ≥80 toE canis, E equi, orE risticiiantigens. Of the 48 IFA seroreactors, 44 dogs were seroreactive toE canis, 21 toE equi, and 0 toE risticii.Seventeen dogs reacted to bothE canisandE equiantigens. There was concordance ofE canisIFA and western immunoblot (WI) test results for 36/44 dogs. Because of cross‐reactivity ofE canissera withE equiantigens, WI was of less utility to confirmE equiexposure. After elimination ofE canisseroreactors, there was concordance of 2/4E equiIFA and WI test results. Based upon a retrospective review of medical records, ehrlichiosis was diagnosed in 10/48 (21%) IFA seroreactive dogs, 9 of which were confirmed positive by WI. Of the remaining 38 IFA seroreactors, 29 also were confirmed byE canisorE equiWI. These results indicate that (1) ehrlichiosis was not diagnosed in the majority of serologically confirmed cases, (2) based uponE canisandE equiWI analysis, IFA testing was not specific (21% false positive), (3)E canissera cross‐react withE equiantigens, and (4) serologic evidence ofE risticiiinfection was lacking in the dog population studied.}, number={1}, journal={Journal of Veterinary Internal Medicine}, author={Suksawat, J. and Hegarty, B. C. and Breitschwerdt, Edward}, year={2000}, pages={50–55} }