@article{neupane_hegarty_marr_maggi_birkenheuer_breitschwerdt_2018, title={Evaluation of cell culture-grown Bartonella antigens in immunofluorescent antibody assays for the serological diagnosis of bartonellosis in dogs}, volume={32}, ISSN={0891-6640}, url={http://dx.doi.org/10.1111/jvim.15301}, DOI={10.1111/jvim.15301}, abstractNote={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.}, number={6}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Neupane, Pradeep and Hegarty, Barbara C. and Marr, Henry S. and Maggi, Ricardo G. and Birkenheuer, Adam J. and Breitschwerdt, Edward B.}, year={2018}, month={Oct}, pages={1958–1964} } @article{o’nion_montilla_qurollo_maggi_hegarty_tornquist_breitschwerdt_2015, title={Potentially Novel Ehrlichia Species in Horses, Nicaragua}, volume={21}, ISSN={1080-6040 1080-6059}, url={http://dx.doi.org/10.3201/eid2102.140290}, DOI={10.3201/eid2102.140290}, abstractNote={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.}, number={2}, journal={Emerging Infectious Diseases}, publisher={Centers for Disease Control and Prevention (CDC)}, author={O’Nion, Victoria L. and Montilla, Hernan J. and Qurollo, Barbara A. and Maggi, Ricardo G. and Hegarty, Barbara C. and Tornquist, Susan J. and Breitschwerdt, Edward B.}, year={2015}, month={Feb}, pages={335–338} } @article{hegarty_qurollo_thomas_park_chandrashekar_beall_thatcher_breitschwerdt_2015, title={Serological and molecular analysis of feline vector-borne anaplasmosis and ehrlichiosis using species-specific peptides and PCR}, volume={8}, ISSN={1756-3305}, url={http://dx.doi.org/10.1186/s13071-015-0929-8}, DOI={10.1186/s13071-015-0929-8}, abstractNote={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. 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. 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. 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.}, number={1}, journal={Parasites & Vectors}, publisher={Springer Science and Business Media LLC}, author={Hegarty, Barbara C. and Qurollo, Barbara A. and Thomas, Brittany and Park, Karen and Chandrashekar, Ramaswamy and Beall, Melissa J. and Thatcher, Brendon and Breitschwerdt, Edward B.}, year={2015}, month={Jun}, pages={320} } @article{arraga-alvarado_qurollo_parra_berrueta_hegarty_breitschwerdt_2014, title={Case Report: Molecular Evidence of Anaplasma platys Infection in Two Women from Venezuela}, volume={91}, ISSN={["1476-1645"]}, DOI={10.4269/ajtmh.14-0372}, abstractNote={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.}, number={6}, journal={AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE}, author={Arraga-Alvarado, Cruz M. and Qurollo, Barbara A. and Parra, Omaira C. and Berrueta, Maribel A. and Hegarty, Barbara C. and Breitschwerdt, Edward B.}, year={2014}, month={Dec}, pages={1161–1165} } @article{lanza-perea_zieger_qurollo_hegarty_pultorak_kumthekar_bruhl-day_breitschwerdt_2014, title={Intraoperative Bleeding in Dogs from Grenada Seroreactive to Anaplasma platys and Ehrlichia canis}, volume={28}, ISSN={0891-6640}, url={http://dx.doi.org/10.1111/jvim.12442}, DOI={10.1111/jvim.12442}, abstractNote={Background Frequent exposure of Grenadian dogs to Rhipicephalus sanguineus results in Anaplasma platys, and Ehrlichia canis seroreactivity. During elective surgeries, substantial intraoperative hemorrhage occurs in some seroreactive dogs. Objectives To assess hemostatic parameters and bleeding tendencies as well as prevalence of PCR positivity in apparently healthy A. platys and E. canis seroreactive and seronegative free‐roaming dogs from Grenada. Animals Forty‐seven elective surgery dogs allocated to 4 groups: Seronegative control (n = 12), A. platys (n = 10), E. canis (n = 14) and A. platys, and E. canis (n = 11) seroreactive. Methods Preoperatively, hemostasis was assessed by platelet count, prothrombin time, activated partial thromboplastin time, and buccal mucosal bleeding time. Intra‐ and postoperative bleeding scores were subjectively assigned. Blood, spleen, bone marrow, and lymph node aspirates were tested by PCR. Results Bleeding scores in dogs coseroreactive for A. platys and E. canis were higher (P = .015) than those of seronegative dogs. A. platys DNA was amplified from 7/21 (33%) A. platys seroreactive dogs and from 1 E. canis seroreactive dog; E. canis DNA was amplified from 21/25 (84%) E. canis seroreactive dogs. E. canis DNA was amplified most often from blood, whereas A. platys DNA was amplified most often from bone marrow. Conclusions and Clinical Importance Apparently healthy, free‐roaming dogs coseropositive for A. platys and E. canis may have increased intraoperative bleeding tendencies despite normal hemostatic parameters. Future investigations should explore the potential for vascular injury as a cause for bleeding in these dogs. Improved tick control is needed for dogs in Grenada.}, number={6}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Lanza-Perea, M. and Zieger, U. and Qurollo, B.A. and Hegarty, B.C. and Pultorak, E.L. and Kumthekar, S. and Bruhl-Day, R. and Breitschwerdt, E.B.}, year={2014}, month={Oct}, pages={1702–1707} } @article{breitschwerdt_hegarty_qurollo_saito_maggi_blanton_bouyer_2014, title={Intravascular persistence of Anaplasma platys, Ehrlichia chaffeensis, and Ehrlichia ewingii DNA in the blood of a dog and two family members}, volume={7}, ISSN={1756-3305}, url={http://dx.doi.org/10.1186/1756-3305-7-298}, DOI={10.1186/1756-3305-7-298}, abstractNote={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.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.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.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.}, number={1}, journal={Parasites & Vectors}, publisher={Springer Nature}, author={Breitschwerdt, Edward B and Hegarty, Barbara C and Qurollo, Barbara A and Saito, Tais B and Maggi, Ricardo G and Blanton, Lucas S and Bouyer, Donald H}, year={2014}, pages={298} } @article{yancey_hegarty_qurollo_levy_birkenheuer_weber_diniz_breitschwerdt_2014, title={Regional Seroreactivity and Vector-Borne Disease Co-Exposures in Dogs in the United States from 2004–2010: Utility of Canine Surveillance}, volume={14}, ISSN={1530-3667 1557-7759}, url={http://dx.doi.org/10.1089/vbz.2014.1592}, DOI={10.1089/vbz.2014.1592}, abstractNote={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.}, number={10}, journal={Vector-Borne and Zoonotic Diseases}, publisher={Mary Ann Liebert Inc}, author={Yancey, Caroline B. and Hegarty, Barbara C. and Qurollo, Barbara A. and Levy, Michael G. and Birkenheuer, Adam J. and Weber, David J. and Diniz, Pedro P.V.P. and Breitschwerdt, Edward B.}, year={2014}, month={Oct}, pages={724–732} } @article{mascarelli_maggi_hopkins_mozayeni_trull_bradley_hegarty_breitschwerdt_2013, title={Bartonella henselae infection in a family experiencing neurological and neurocognitive abnormalities after woodlouse hunter spider bites}, volume={6}, ISSN={1756-3305}, url={http://dx.doi.org/10.1186/1756-3305-6-98}, DOI={10.1186/1756-3305-6-98}, abstractNote={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.}, number={1}, journal={Parasites & Vectors}, publisher={Springer Nature}, author={Mascarelli, Patricia E and Maggi, Ricardo G and Hopkins, Sarah and Mozayeni, B Robert and Trull, Chelsea L and Bradley, Julie M and Hegarty, Barbara C and Breitschwerdt, Edward B}, year={2013}, pages={98} } @article{balakrishnan_cherry_linder_pierce_sontakke_hegarty_bradley_maggi_breitschwerdt_2013, title={Experimental infection of dogs with Bartonella henselae and Bartonella vinsonii subsp. berkhoffii}, volume={156}, ISSN={0165-2427}, url={http://dx.doi.org/10.1016/j.vetimm.2013.09.007}, DOI={10.1016/j.vetimm.2013.09.007}, abstractNote={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.}, number={1-2}, journal={Veterinary Immunology and Immunopathology}, publisher={Elsevier BV}, author={Balakrishnan, Nandhakumar and Cherry, Natalie A. and Linder, Keith E. and Pierce, Eric and Sontakke, Neal and Hegarty, Barbara C. and Bradley, Julie M. and Maggi, Ricardo G. and Breitschwerdt, Edward B.}, year={2013}, month={Nov}, pages={153–158} } @article{hegarty_maggi_koskinen_beall_eberts_chandrashekar_breitschwerdt_2012, title={Ehrlichia muris Infection in a Dog from Minnesota}, volume={26}, ISSN={0891-6640}, url={http://dx.doi.org/10.1111/j.1939-1676.2012.00968.x}, DOI={10.1111/j.1939-1676.2012.00968.x}, abstractNote={A 6-year-old-male Labrador Retriever from northern Minnesota was examined by a veterinarian in June 2011 because of decreased activity, reluctance to climb stairs, and a history of having 2 attached ticks, approximately 2 months earlier. The dog was febrile (rectal temperature 103.5°F) and had a stiff gait, particularly noticeable in the front legs. On palpation, there was pain in both elbow joints. Because of the high prevalence of tick-borne arthropathies in the practice area, Lyme borreliosis or anaplasmosis was suspected. Because a commercially available in-house assay1 was strongly positive for Anaplasma species antibodies, canine anaplasmosis was diagnosed serologically and doxycycline was administered at a dosage of 6 mg/kg PO q12h for 21 days. Carprofen (2 mg/kg PO q24h–q12h) was dispensed for control of pain as needed and topical fipronil was dispensed for tick control. When re-examined 5 days later, overall activity was decreased, the dog frequently licked the paws and remained stiff, but the owner had not administered carprofen. Rectal temperature was 102.2°F. Amoxicillin (11 mg/kg PO q12h for 10 days) and Fortiflora were added to the treatment regimen. When re-examined in July, the dog had returned to normal activities and normal mobility. On September 7, 2011, a second veterinarian collaborating with the Intracellular Pathogens Research Laboratory (IPRL) examined the dog because of decreased appetite, lethargy, and recurrent bouts of vomiting. Rectal temperature was 103.3°F. SNAP 4Dx again indicated the presence of Anaplasma species antibodies. The dog was thrombocytopenic (platelet count, 132,000/μL; reference range, 160,000–525,000/μL), according to an in-house platelet count, but a CBC and serum biochemical profile performed by a commercial diagnostic laboratory2 on a blood sample drawn the same day identified no hematologic or serum biochemical abnormalities (platelet count, 243,000/μL; reference range, 140,000–540,000/μL). Initial treatment included 1 dose of oxytetracycline (1 mg/kg, IV). Doxycycline was dispensed with instructions to administer 8 mg/kg PO q12h for 30 days along with a digestive support supplement.3 Blood and serum samples collected on September 7, 2011 were sent to the IPRL for serological and PCR testing for Anaplasma and Ehrlichia spp. Anaplasma species antibodies were confirmed using SNAP 4Dx. PCR amplification was performed using previously described GEP 16S Ehrlichia genus primers and species-specific primers for Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia ewingii, Anaplasma phagocytophilum, and Anaplasma platys.1 An amplicon was obtained with the Ehrlichia genus primers, but amplification using the above-mentioned species-specific primers, including A. phagocytophilum, did not generate amplicons. Sequencing4 of the Ehrlichia genus amplicon resulted in the highest DNA similarity (100%) to Ehrlichia muris (361 of 361 base pairs when compared to GenBank Accession number NR_025962, a strain from a mouse in Japan). After designing a 16S rDNA, E. muris primer pair Emu58 (sense): 5′ ATAGCTACCCATAGCTTTTTTAGCTATAGGTT 3′ and SEP (anti-sense): 5′ CTTCTRTRGGTACCGTCATTATCTTCCCY 3′ (as forward and reverse primers respectively), a 395 base pair amplicon (expected amplicon size) was obtained from the September 7, 2011 blood sample. The dog was examined on September 23, 2011 because of vomiting and diarrhea that was temporally associated with administration of doxycycline. A fecal flotation was negative for parasite ova. Clostridium enteritis was diagnosed by the attending veterinarian using fecal smear examination. Doxycycline was discontinued and amoxicillin (20 mg/kg POq12h for 14 days), metronidazole (20 mg/kg PO q12h for 14 days), and metoclopramide (0.9 mg/kg PO q24 for 5 days) were dispensed. Within 3 days, the vomiting and diarrhea had resolved and the dog was acting normally. PCR amplification5 using Ehrlichia genus and E. muris-specific primers from a September 27, 2011 blood sample were negative, no additional doxycycline was dispensed. On October 29, 2011, the dog was somewhat less active, had bloody urine, and occasional blood clots were observed dripping from the prepuce. The prostate gland was slightly enlarged, nonpainful, and symmetrical. The packed cell volume was 48%, the platelet count was 450,000/μL (reference range, 166,000–575,000/μL), and hypoglobulinemia (2.1 g/dL; reference range, 2.5–4.5 g/dL) was present. Urine was alkaline (pH8.0) with 3+ proteinuria and specific gravity 1.040, and hematuria was present. Radiographs of the abdomen disclosed no abnormalties. Cephalexin (20 mg/kg PO q8h for 10 days) was dispensed for presumptive urinary tract infection. Despite treatment with cephalexin, hematuria persisted. On November 2, 2011, the platelet count was 118,500/μL and packed cell volume was 50%. Urine specific gravity was 1.040 and sediment examination identified too numerous to count erythrocytes and occasional leukocytes. Because of the thrombocytopenia and because the dog had not completed the 30-day course of doxycycline because of vomiting and diarrhea in September, doxycycline again was administered (5 mg/kg PO q12h for 30 days). In addition, the dog was referred for an abdominal ultrasound examination. No ultrasonographic structural abnormalities were found within the urinary tract. The hematuria resolved and the dog did not experience any additional medical problems. On November 16, 2011, post-treatment blood sample for PCR and convalescent serum for testing against a panel of vector-borne pathogens were submitted to the IPRL. Again, seroreactivity to the Anaplasma peptide was the only SNAP 4Dx finding. By IFA testing, there was no antibody reactivity at a titer of 1 : 16 (testing scale, 1 : 16–1 : 8,192) to Babesia canis, Bartonella henselae, Bartonella vinsonii subsp. berkhoffii, E. canis, and Rickettsia rickettsii antigens. Ehrlichia genus and E. muris-specific PCR were negative. In addition, PCR amplification was retrospectively performed for the 3 sample collection dates using previously described groEL Ehrlichia genus primers.2 A GroEL PCR amplicon was obtained from the blood sample collected on September 7, 2011, but not from blood samples collected on September 27, 2011 and November 16, 2011. Sequence analysis of the GroEL PCR amplicon showed 100% homology (547/547 bp) with E. muris (GenBank accession number AF210459, from the E. muris type strain AS145). The E. muris 16S and GroEL sequences derived from this dog have been deposited into the GenBank data base under the accession numbers JQ10629 and JQ10630, respectively. Sequence phylogeny analyses of the 16S and GroEL gene sequences conducted with Molecular Evolutionary Genetics Analysis software supported a close phylogenetic relationship with E. muris. (Fig 1). In collaboration with 2 veterinary clinicians (Drs Koskinen and Eberts) in Minnesota, who routinely examine dogs with acute, febrile illness, we were able to provide the first molecular diagnostic evidence to support a potential role for E. muris as a pathogen in dogs from the United States. In 2 previously published studies, seroreactivity to E. canis-derived peptides using SNAP 3DX or 4DX assays in dogs from the Minnesota was unexpectedly more prevalent than expected based on tick vectors found in this region.1, 3 Exposure to Rhipicephalus sangineus, the vector for E. canis, and Ambylomma americanum, the vector for E. chaffeensis and E. ewingii, occurs infrequently in colder regions of the United States, therefore infection with another bacterial pathogen, potentially transmitted by Ixodes scapularis, a tick that is plentiful throughout this region, was suspected. In addition, E. canis and E. chaffeensis are the only 2 recognized Ehrlichia species in North America that are known to induce seroreactivity to the E. canis peptides used in the SNAP 3Dx and 4Dx assays.4, 5 In a previous study, neither E. canis nor E. chaffeensis DNA was amplified from Minnesota dogs, whereas the B. burgdorferi and A. phagocytophilum seroprevalence was 55%, indicating frequent exposure to I. scapularis.1 Based upon serology, the dog in this study had been exposed to A. phagocytophilum, supporting exposure to I. scapularis. In 2011, after the publications by Bowman and Beall,1, 3 human infection with an E. muris-like agent was reported for the first time in the medical literature in patients from Wisconsin.6 In addition, Telford et al. retrospectively documented the presence of E. muris DNA in I. scapularis ticks collected in the 1990s from northern Wisconsin.7 Collectively, these observations suggested that pet dogs exposed to ticks in this region also might be infected with this novel pathogen within the genus Ehrlichia. Potentially, cross-reacting antibodies resulting from exposure to another genus of bacteria provided a plausible explanation for the high SNAP E. canis seroreactivity in dogs from this region, but it seemed more likely that an organism within the genus Ehrlichia might be responsible for the seemingly disparate seroepidemiological findings. Interestingly, the E. muris-infected dog in this study never seroconverted to the E. canis peptides using SNAP 4Dx or E. canis IFA assays. Thus, some E. muris-infected dogs may not seroconvert, particularly if antibiotic treatment is started very early in the course of infection, or alternatively, E. muris may not be responsible for the sero-epidemiologic observations in previous studies from Minnesota.1, 3 In the context of human illness, the 4 patients described to date, most of whom have been immunocompromised, presented with fever, headache, thrombocytopenia, lymphopenia, and increased liver enzyme activities.6 Experimentally, E. muris-infected mice developed splenomegaly and lymphadenopathy.8 Although the role of E. muris in the overall pathogenesis of disease manifestations reported in this dog between June and November 2011 is impossible to assess with the available clinical, serologic and microbiological data, fever and thrombocytopenia are features of illness in human patients. Moreover, the expected therapeutic response to doxycycline treatment in dogs infected with A. phagocytophilum is generally dramatic, with rapid resolution of disease manifestations occurring generally within 24–48 hours.9, 10 Given the fact that this dog remained stiff and inactive for at least 5 days after initially being treated with doxycycline in June, it is possible that the dog was co-infected with A. phagocytophilum and E. muris at the time of the initial evaluation and that the E. muris infection persisted until documented in September 2011. Alternatively, A. phagoctyophilum may have been solely responsible for the June presentation, with E. muris transmission occurring in early September, just before onset of the second documented febrile episode. Recently, to further complicate clinical interpretation of disease outcomes, chronic infection with A. phagocytophilum has been documented in experimentally infected dogs in which treatment with doxycycline did not eliminate the infection.11 Despite this research observation, to date, molecular evidence supporting chronic canine anaplasmosis in naturally infected dogs is lacking in the literature from North American and European regions, where A. phagocytophilum transmission is endemic, and efforts to PCR amplify A. phagocytophilum DNA from 3 blood samples from the dog of this study were not successful. Whether E. muris causes an acute, short duration infection in dogs or is capable of inducing chronic long-lasting (months to years) infection in dogs, as occurs with other Ehrlichia spp., such as E. canis and E. ewingii, remains unknown. Historically, because Ehlrichia spp. could only be cultured using cell culture systems in a research setting, the advent of diagnostic DNA testing has greatly facilitated enhanced understanding of the host range and the pathogenic potential of these obligate intracellular bacteria. Ehrlichia muris was first isolated from rodents in Japan in the early 1990s.12 An experimental infection study involving 2 dogs failed to demonstrate evidence of transmission and the dogs did not develop signs of disease.8 Subsequently, monocytic Ehrlichia sp. DNA with sequence homology to E. muris was PCR-amplified from Ixodes persulcatus and Ixodes ricinus ticks found in Eastern Europe.13, 14 Unlike the DNA sequences from human patients, which were reported as E. muris-like,6 the 16S rDNA and rpoB sequences obtained from the dog in this study were 100% homologous to the E. muris type strain from Japanese rodents, and differed from the sequences reported from human patients in Wisconsin. The routine diagnostic use of PCR amplification and DNA sequencing practices are likely to identify other as yet unknown Ehrlichia sp. and an expanded host range in which infection with various Ehrlichia sp. occurs.}, number={5}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Hegarty, B.C. and Maggi, R.G. and Koskinen, P. and Beall, M.J. and Eberts, M. and Chandrashekar, R. and Breitschwerdt, E.B.}, year={2012}, month={Jul}, pages={1217–1220} } @article{cherry_maggi_rossmeisl_hegarty_breitschwerdt_2011, title={Ecological Diversity ofBartonellaSpecies Infection Among Dogs and Their Owner in Virginia}, volume={11}, ISSN={1530-3667 1557-7759}, url={http://dx.doi.org/10.1089/vbz.2010.0201}, DOI={10.1089/vbz.2010.0201}, abstractNote={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.}, number={11}, journal={Vector-Borne and Zoonotic Diseases}, publisher={Mary Ann Liebert Inc}, author={Cherry, Natalie A. and Maggi, Ricardo G. and Rossmeisl, John H. and Hegarty, Barbara C. and Breitschwerdt, Edward B.}, year={2011}, month={Nov}, pages={1425–1432} } @article{breitschwerdt_mascarelli_schweickert_maggi_hegarty_bradley_woods_2011, title={Hallucinations, Sensory Neuropathy, and Peripheral Visual Deficits in a Young Woman Infected with Bartonella koehlerae}, volume={49}, ISSN={0095-1137 1098-660X}, url={http://dx.doi.org/10.1128/JCM.00833-11}, DOI={10.1128/jcm.00833-11}, abstractNote={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.}, number={9}, journal={Journal of Clinical Microbiology}, publisher={American Society for Microbiology}, author={Breitschwerdt, Edward B. and Mascarelli, Patricia E. and Schweickert, Lori A. and Maggi, Ricardo G. and Hegarty, Barbara C. and Bradley, Julie M. and Woods, Christopher W.}, year={2011}, month={Aug}, pages={3415–3417} } @article{breitschwerdt_hegarty_maggi_lantos_aslett_bradley_2011, title={Rickettsia rickettsii transmission by a lone star tick, North Carolina}, volume={17}, number={5}, journal={Emerging Infectious Diseases}, author={Breitschwerdt, E. B. and Hegarty, B. C. and Maggi, R. G. and Lantos, P. M. and Aslett, D. M. and Bradley, J. M.}, year={2011}, pages={873–875} } @article{breitschwerdt_maggi_lantos_woods_hegarty_bradley_2010, title={Bartonella vinsonii subsp berkhoffii and Bartonella henselae bacteremia in a father and daughter with neurological disease}, volume={3}, journal={Parasites & Vectors}, author={Breitschwerdt, E. B. and Maggi, R. G. and Lantos, P. M. and Woods, C. W. and Hegarty, B. C. and Bradley, J. M.}, year={2010} } @article{diniz_beall_omark_chandrashekar_daniluk_cyr_koterski_robbins_lalo_hegarty_et al._2010, title={High Prevalence of Tick-Borne Pathogens in Dogs from an Indian Reservation in Northeastern Arizona}, volume={10}, ISSN={1530-3667 1557-7759}, url={http://dx.doi.org/10.1089/vbz.2008.0184}, DOI={10.1089/vbz.2008.0184}, abstractNote={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.}, number={2}, journal={Vector-Borne and Zoonotic Diseases}, publisher={Mary Ann Liebert Inc}, author={Diniz, Pedro Paulo V.P. and Beall, Melissa J. and Omark, Karina and Chandrashekar, Ramaswamy and Daniluk, Daryn A. and Cyr, Katie E. and Koterski, James F. and Robbins, Richard G. and Lalo, Pamela G. and Hegarty, Barbara C. and et al.}, year={2010}, month={Mar}, pages={117–123} } @article{breitschwerdt_maggi_mozayeni_hegarty_bradley_mascarelli_2010, title={PCR amplification of Bartonella koehlerae from human blood and enrichment blood cultures}, volume={3}, journal={Parasites & Vectors}, author={Breitschwerdt, E. B. and Maggi, R. G. and Mozayeni, B. R. and Hegarty, B. C. and Bradley, J. M. and Mascarelli, P. E.}, year={2010} } @article{hegarty_diniz_bradley_lorentzen_breitschwerdt_2009, title={Clinical Relevance of Annual Screening Using a Commercial Enzyme-Linked Immunosorbent Assay (SNAP 3Dx) for Canine Ehrlichiosis}, volume={45}, ISSN={["1547-3317"]}, DOI={10.5326/0450118}, abstractNote={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.}, number={3}, journal={JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION}, author={Hegarty, Barbara C. and Diniz, Pedro Paulo Vissotto de Paiva and Bradley, Julie M. and Lorentzen, Leif and Breitschwerdt, Edward}, year={2009}, pages={118–124} } @article{lappin_breitschwerdt_brewer_hawley_hegarty_radecki_2009, title={Prevalence of Bartonella species antibodies and Bartonella species DNA in the blood of cats with and without fever}, volume={11}, ISSN={1098-612X 1532-2750}, url={http://dx.doi.org/10.1016/j.jfms.2008.06.005}, DOI={10.1016/j.jfms.2008.06.005}, abstractNote={The purpose of this study was to determine whether there are associations between Bartonella species antibody (enzyme-linked immunosorbent assay (ELISA) and Western blot (WB)) and polymerase chain reaction assay results in cats with and without fever. Afebrile control cats (39/93; 42.0%) were more likely to have Bartonella species antibodies than cats with fever (29/93; 31.2%). The difference in prevalence of Bartonella species deoxyribonucleic acid (DNA) in blood of cats with fever (14/81; 17.3%) as compared to afebrile control cats (6/81; 7.4%) approached statistical significance (P=0.0571). Bartonella species ELISA or WB results frequently did not correlate to the presence or absence of Bartonella species DNA in blood. The results of this study indicate that in cats, Bartonella species antibody tests cannot predict whether fever is due to Bartonella species infection and should not be used to determine the Bartonella species infection status.}, number={2}, journal={Journal of Feline Medicine and Surgery}, publisher={SAGE Publications}, author={Lappin, Michael R. and Breitschwerdt, Edward and Brewer, Melissa and Hawley, Jennifer and Hegarty, Barbara and Radecki, Steven}, year={2009}, month={Feb}, pages={141–148} } @article{kidd_maggi_diniz_hegarty_tucker_breitschwerdt_2008, title={Evaluation of conventional and real-time PCR assays for detection and differentiation of Spotted Fever Group Rickettsia in dog blood}, volume={129}, ISSN={0378-1135}, url={http://dx.doi.org/10.1016/j.vetmic.2007.11.035}, DOI={10.1016/j.vetmic.2007.11.035}, abstractNote={Spotted Fever Group Rickettsia is important cause of emerging and re-emerging infectious disease in people and dogs. Importantly, dogs can serve as sentinels for disease in people. Sensitive and specific diagnostic tests that differentiate among species of infecting Rickettsia are needed. The objective of this study was to develop a sensitive and specific PCR that differentiates SFG Rickettsia infecting dog blood. Conventional and real-time PCR assays were developed using primers that targeted a small region of the ompA gene. Their sensitivity, determined by testing a cloned target sequence in the presence of host DNA, was 15–30 and 5 copies of DNA, respectively. Testing of Rickettsia cultures and analysis of Rickettsia gene sequences deposited in GenBank verified DNA could be amplified and used to differentiate species. DNA from the blood of infected dogs was also tested. Importantly, Rickettsia DNA was detected before seroconversion in some dogs. The species of infecting Rickettsia was also identified. We conclude these assays may assist in the timely diagnosis of infection with SFG Rickettsia. They may also facilitate the discovery of novel SFG Rickettsia infecting dogs, and in the investigation of dogs as sentinels for emerging rickettsioses.}, number={3-4}, journal={Veterinary Microbiology}, publisher={Elsevier BV}, author={Kidd, L. and Maggi, R. and Diniz, P.P.V.P. and Hegarty, B. and Tucker, M. and Breitschwerdt, E.}, year={2008}, month={Jun}, pages={294–303} } @article{cadenas_bradley_maggi_takara_hegarty_breitschwerdt_2008, title={Molecular characterization of Bartonella vinsonii subsp berkhoffii genotype III}, volume={46}, DOI={10.1128/JCM.62456-07}, number={5}, journal={Journal of Clinical Microbiology}, author={Cadenas, M. B. and Bradley, J. and Maggi, Ricardo and Takara, M. and Hegarty, B. C. and Breitschwerdt, Edward}, year={2008}, pages={1858–1860} } @article{beall_chandrashekar_eberts_cyr_diniz_mainville_hegarty_crawford_breitschwerdt_2008, title={Serological and Molecular Prevalence of Borrelia burgdorferi, Anaplasma phagocytophilum, and Ehrlichia Species in Dogs from Minnesota}, volume={8}, ISSN={1530-3667 1557-7759}, url={http://dx.doi.org/10.1089/vbz.2007.0236}, DOI={10.1089/vbz.2007.0236}, abstractNote={A population of 731 naturally exposed pet dogs examined at a private practice in Baxter, Minnesota, an area endemic for Lyme disease and anaplasmosis, was tested by serological and molecular methods for evidence of exposure to or infection with selected vector-borne pathogens. Serum samples were tested by enzyme-linked immunosorbent assay (ELISA) for Anaplasma phagocytophilum, Borrelia burgdorferi, and Ehrlichia canis antibodies and for Dirofilaria immitis antigen. Blood samples from 273 dogs were also analyzed by polymerase chain reaction (PCR) for Anaplasma and Ehrlichia species DNA. Based on the owner history and the attending veterinarian's physical examination findings, dogs exhibiting illness compatible with anaplasmosis or borreliosis were considered clinical cases, and their results were compared to the healthy dog population. Antibodies to only A. phagocytophilum were detected in 217 (29%) dogs; to only B. burgdorferi, in 80 (11%) dogs; and seroreactivity to both organisms, in 188 (25%) dogs. Of 89 suspected cases of canine anaplasmosis or borreliosis, A. phagocytophilum or B. burgdorferi antibodies were detected in 22 dogs (25%) and 8 dogs (9%) respectively, whereas antibodies to both organisms were found in 38 dogs (43%). Ehrlichia canis antibodies and D. immitis antigen were each detected in 11 (1.5%) dogs. Anaplasma phagocytophilum DNA was amplified from 7 of 222 (3%) healthy dogs and 19 of 51 (37%) clinical cases. Seroreactivity to both A. phagocytophilum and B. burgdorferi was detected more frequently in suspected cases of anaplasmosis and/or borreliosis than seroreactivity to either organism alone. Based on PCR testing, A. phagocytophilum DNA was more prevalent in suspected cases of anaplasmosis or borreliosis than in healthy dogs from the same region.}, number={4}, journal={Vector-Borne and Zoonotic Diseases}, publisher={Mary Ann Liebert Inc}, author={Beall, Melissa J. and Chandrashekar, Ramaswamy and Eberts, Matthew D. and Cyr, Katie E. and Diniz, Pedro Paulo V.P. and Mainville, Celine and Hegarty, Barbara C. and Crawford, John M. and Breitschwerdt, Edward B.}, year={2008}, month={Aug}, pages={455–464} } @article{breitschwerdt_maggi_duncan_nicholson_hegarty_woods_2007, title={BartonellaSpecies in Blood of Immunocompetent Persons with Animal and Arthropod Contact}, volume={13}, ISSN={1080-6040 1080-6059}, url={http://dx.doi.org/10.3201/eid1306.061337}, DOI={10.3201/eid1306.061337}, abstractNote={Using PCR in conjunction with pre-enrichment culture, we detected Bartonella henselae and B. vinsonii subspecies berkhoffii in the blood of 14 immunocompetent persons who had frequent animal contact and arthropod exposure.}, number={6}, journal={Emerging Infectious Diseases}, publisher={Centers for Disease Control and Prevention (CDC)}, author={Breitschwerdt, Edward B. and Maggi, Ricardo G. and Duncan, Ashlee W. and Nicholson, William L. and Hegarty, Barbara C. and Woods, Christopher W.}, year={2007}, month={Jun}, pages={938–941} } @article{vissotto de paiva diniz_maggi_schwartz_cadenas_bradley_hegarty_breitschwerdt_2007, title={Canine bartonellosis: serological and molecular prevalence in Brazil and evidence of co-infection with Bartonella henselae and Bartonella vinsonii subsp berkhoffii}, volume={38}, ISSN={["0928-4249"]}, DOI={10.1051/vetres:2007023}, abstractNote={The purpose of this study was to determine the serological and molecular prevalence of Bartonella spp. infection in a sick dog population from Brazil. At the São Paulo State University Veterinary Teaching Hospital in Botucatu, 198 consecutive dogs with clinicopathological abnormalities consistent with tick-borne infections were sampled. Antibodies to Bartonella henselae and Bartonella vinsonii subsp. berkhoffii were detected in 2.0% (4/197) and 1.5% (3/197) of the dogs, respectively. Using 16S-23S rRNA intergenic transcribed spacer (ITS) primers, Bartonella DNA was amplified from only 1/198 blood samples. Bartonella seroreactive and/or PCR positive blood samples (n=8) were inoculated into a liquid pre-enrichment growth medium (BAPGM) and subsequently sub-inoculated onto BAPGM/blood-agar plates. PCR targeting the ITS region, pap31 and rpoB genes amplified B. henselae from the blood and/or isolates of the PCR positive dog (ITS: DQ346666; pap31 gene: DQ351240; rpoB: EF196806). B. henselae and B. vinsonii subsp. berkhoffii (pap31: DQ906160; rpoB: EF196805) co-infection was found in one of the B. vinsonii subsp. berkhoffii seroreactive dogs. We conclude that dogs in this study population were infrequently exposed to or infected with a Bartonella species. The B. henselae and B. vinsonii subsp. berkhoffii strains identified in this study are genetically similar to strains isolated from septicemic cats, dogs, coyotes and human beings from other parts of the world. To our knowledge, these isolates provide the first Brazilian DNA sequences from these Bartonella species and the first evidence of Bartonella co-infection in dogs.}, number={5}, journal={VETERINARY RESEARCH}, author={Vissotto De Paiva Diniz, Pedro Paulo and Maggi, Ricardo Guillermo and Schwartz, Denise Saretta and Cadenas, Maria Belen and Bradley, Julie Meredith and Hegarty, Barbara and Breitschwerdt, Edward Bealmear}, year={2007}, pages={697–710} } @article{eddlestone_diniz_neer_gaunt_corstvet_cho_hosgood_hegarty_breitschwerdt_2007, title={Doxycycline Clearance of Experimentally Induced Chronic Ehrlichia Canis Infection in Dogs}, volume={21}, ISSN={0891-6640}, url={http://dx.doi.org/10.1892/07-061.1}, DOI={10.1892/07-061.1}, abstractNote={Ineffective clearance of Ehrlichia canis after doxycycline administration has been reported despite the fact that the recommended treatment for canine ehrlichiosis is doxycycline. The effectiveness of doxycycline in clearing E canis infection from the blood and tissues of dogs requires additional evaluation.Doxycycline (5 mg/kg PO q12h), administered for 4 weeks, will eliminate E canis infection from the blood and tissues of experimentally infected dogs.Fifteen Walker hound-mixed breed dogs were inoculated subcutaneously with E canis-infected canine histiocytic cells 4 months before doxycycline treatment.Four dogs were treated with doxycycline (5 mg/kg PO q12h for 3 weeks), 5 dogs were treated with doxycycline at the same dosage for 4 weeks, and 5 control dogs were not treated. Dexamethasone (0.4 mg/kg i.v.) was given after treatment to precipitate recrudescence of any remaining E canis organisms. Platelet counts, anti-E canis immunofluorescent antibodies, and polymerase chain reaction (PCR) detection of E canis deoxyribonucleic acid (DNA) in blood and tissues were evaluated.E canis DNA was not detected in the blood and tissues of doxycycline-treated dogs after treatment. Platelet counts were within reference intervals, and E canis antibodies decreased. Spontaneous clearance of E canis infection occurred in 2 of 5 control dogs. Three control dogs had E canis DNA detected in blood and tissues, platelet counts remained low or within the reference interval, and E canis antibodies remained high.As administered in this study, doxycycline cleared E canis from the blood and tissues of experimentally infected dogs.}, number={6}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Eddlestone, S.M. and Diniz, P.P.V.P. and Neer, T.M. and Gaunt, S.D. and Corstvet, R. and Cho, D. and Hosgood, G. and Hegarty, B. and Breitschwerdt, E.B.}, year={2007}, pages={1237–1242} } @article{jones_valenzisi_sontakke_sprayberry_maggi_hegarty_breitschwerdt_2007, title={Use of an insect cell culture growth medium to isolate bacteria from horses with effusive, fibrinous pericarditis: A preliminary study}, volume={121}, ISSN={0378-1135}, url={http://dx.doi.org/10.1016/j.vetmic.2006.11.024}, DOI={10.1016/j.vetmic.2006.11.024}, abstractNote={Effusive, fibrinous pericarditis is an uncommon disease entity in horses. In 2001, pericarditis occurred in conjunction with an epizootic in central Kentucky that was associated with exposure to eastern tent caterpillars (ETCs). Bacterial isolation from equine pericardial fluid samples was attempted using an insect cell culture growth medium (ICCGM). Using previously cultured, stored frozen samples from four horses with fibrinous pericarditis, inoculation of 10% blood agar plates yielded no growth, whereas simultaneous inoculation of ICCGM resulted in the isolation of Proprionibacterium acnes, Staphylococcus equorum, a Streptococcus sp. and Pseudomonas rhodesiae from pericardial fluid samples. A similar or novel caterpillar-associated bacteria was not identified; however, use of an ICCGM might enhance isolation of bacteria from equine pericardial fluid.}, number={1-2}, journal={Veterinary Microbiology}, publisher={Elsevier BV}, author={Jones, Samuel L. and Valenzisi, Amy and Sontakke, Sushama and Sprayberry, Kimberly A. and Maggi, Ricardo and Hegarty, Barbara and Breitschwerdt, Edward}, year={2007}, month={Mar}, pages={177–181} } @article{maggi_chomel_hegarty_henn_breitschwerdt_2006, title={A Bartonella vinsonii berkhoffii typing scheme based upon 16S–23S ITS and Pap31 sequences from dog, coyote, gray fox, and human isolates}, volume={20}, ISSN={0890-8508}, url={http://dx.doi.org/10.1016/j.mcp.2005.11.002}, DOI={10.1016/j.mcp.2005.11.002}, abstractNote={Since the isolation of Bartonella vinsonii subspecies berkhoffii from a dog with endocarditis in 1993, this organism has emerged as an important pathogen in dogs and as an emerging pathogen in people. Current evidence indicates that coyotes, dogs and gray foxes potentially serve as reservoir hosts. Based upon sequence differences within the 16S–23S ITS region and Pap31 gene, we propose a classification scheme that divides B. vinsonii subsp. berkhoffii isolates into four distinct types. Two conserved sequences, of 37 and 18 bp, respectively, are differentially present within the ITS region of each of the four B. vinsonii subsp. berkhoffii types. To date, B. vinsonii berkhoffii types I, II, and III have been identified in the US, type III in Europe and type IV in Canada. Based upon the proposed genotyping scheme, the geographic distribution of B. vinsonii berkhoffii types needs to be more thoroughly delineated in future molecular epidemiological studies involving Bartonella infection in coyotes, dogs, gray foxes, human beings and potentially other animals or in arthropod vectors. Strain typing may help to better define the reservoir potential, carriership patterns, modes of transmission, and geographic distribution for each B. vinsonii berkhoffii type.}, number={2}, journal={Molecular and Cellular Probes}, publisher={Elsevier BV}, author={Maggi, Ricardo G. and Chomel, Bruno and Hegarty, Barbara C. and Henn, Jennifer and Breitschwerdt, Edward B.}, year={2006}, month={Apr}, pages={128–134} } @article{solano-gallego_llull_osso_hegarty_breitschwerdt_2006, title={A serological study of exposure to arthropod-borne pathogens in dogs from northeastern Spain}, volume={37}, ISSN={["1297-9716"]}, DOI={10.1051/vetres:2005054}, abstractNote={There is limited information regarding the prevalence of many vector borne pathogens in Europe and especially in Spanish dogs. We investigated 206 sick and 260 clinically healthy dogs from three different regions in northeastern Spain for antibodies to Rickettsia conorii (Rc), Ehrlichia canis (Ec), Anaplasma phagocytophilum (Ap), Bartonella henselae (Bh), Bartonella vinsonii subsp. berkhoffii (Bvb), Leishmania infantum (Li) and Borrelia burgdorferi (Bb) and for antigen of Dirofilaria immitis (Di). Total prevalences were the following: Rc (56.4%), Li (30%), Ec (16.7%), Bh (16.8%), Ap (11.5%), Bvb (1.07%), Di (0.6%) and Bb (0.6%). Seroprevalences for Rc, Ec, Ap, Bh, and Bvb and Bb and Di antigens were similar among the three different study sites. The Ec seroprevalence, as determined by Snap 3DX, was statistically lower in dogs from Mallorca (0%) than Tarragona (16%) and Barcelona (5%) (P < 0.0001). Detection of Rc antibodies was associated with seroreactivity to Ec and Ap antigens (P = 0.018 and P = 0.002, respectively). IFA Ec antibodies were associated with Ap seroreactivity (P < 0.0001). There was no association between the clinical status, sex, time of the year when samples were collected, life-style or exposure to fleas or ticks and a positive test result for Ec, Bh, Bvb, or Bb antibodies or Di antigens. Li seroreactivity was associated with illness and living outdoors (P < 0.0001, P = 0.029; respectively), Rc seroreactivity with the male gender (P = 0.028) and Ap seroreactivity with living outdoors (P = 0.045). This study indicates that exposure to Rc, Li, Ec or related Ehrlichia spp., Bh and Ap or a related spp., is common whereas Di, Bb and Bvb is uncommon among dogs from the Mediterranean basin. We also provide serological data that suggests the existence of a novel Ehrlichia species on Mallorca island.}, number={2}, journal={VETERINARY RESEARCH}, author={Solano-Gallego, L and Llull, J and Osso, M and Hegarty, B and Breitschwerdt, E}, year={2006}, pages={231–244} } @article{o'connor_hanscom_hegarty_groat_breitschwerdt_2006, title={Comparison of an indirect immunofluorescence assay, western blot analysis, and a commercially available ELISA for detection ofEhrlichia canisantibodies in canine sera}, volume={67}, ISSN={0002-9645}, url={http://dx.doi.org/10.2460/ajvr.67.2.206}, DOI={10.2460/ajvr.67.2.206}, abstractNote={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.}, number={2}, journal={American Journal of Veterinary Research}, publisher={American Veterinary Medical Association (AVMA)}, author={O'Connor, Thomas P. and Hanscom, Jancy L. and Hegarty, Barbara C. and Groat, Randall G. and Breitschwerdt, Edward B.}, year={2006}, month={Feb}, pages={206–210} } @article{hess_english_hegarty_brown_breitschwerdt_2006, title={Experimental Ehrlichia canis infection in the dog does not cause immunosuppression}, volume={109}, ISSN={["1873-2534"]}, DOI={10.1016/j.vetimm.2005.07.027}, abstractNote={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.}, number={1-2}, journal={VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY}, author={Hess, PR and English, RV and Hegarty, BC and Brown, GD and Breitschwerdt, EB}, year={2006}, month={Jan}, pages={117–125} } @article{eddlestone_neer_gaunt_corstvet_gill_hosgood_hegarty_breitschwerdt_2006, title={Failure of imidocarb dipropionate to clear experimentally induced Ehrlichia canis infection in dogs}, volume={20}, number={4}, journal={Journal of Veterinary Internal Medicine}, author={Eddlestone, S. M. and Neer, T. M. and Gaunt, S. D. and Corstvet, R. and Gill, A. and Hosgood, G. and Hegarty, B. and Breitschwerdt, E. B.}, year={2006}, pages={840–844} } @inbook{kidd_hegarty_sexton_breitschwerdt_2006, title={Molecular characterization of Rickettsia rickettsii infecting dogs and people in North Carolina}, volume={1078}, ISBN={1573316016}, booktitle={Century of rickettsiology: emerging, reemerging rickettsioses, molecular diagnostics, and emerging veterinary rickettsioses}, publisher={Oxford: Blackwell Publishing}, author={Kidd, L. and Hegarty, B. and Sexton, D. and Breitschwerdt, E.}, year={2006}, pages={400–409} } @article{solano-gallego_hegarty_espada_llull_breitschwerdt_2006, title={Serological and molecular evidence of exposure to arthropod-borne organisms in cats from northeastern Spain}, volume={118}, ISSN={["0378-1135"]}, DOI={10.1016/j.vetmic.2006.07.010}, abstractNote={One hundred sixty-eight cat sera from Spain were tested for IgG antibodies to Rickettsia conorii (Rc), Ehrlichia canis (Ec), Anaplasma phagocytophilum (Ap) and Bartonella henselae (Bh) antigens using IFA and for FeLV antigen and FIV antibody by ELISA. For 47 whole blood samples, PCR testing was performed for Rickettsia, Ehrlichia and Bartonella. Seroprevalences were: Bh (71.4%), Rc (44%), Ec (11.3%), FeLV (8.5%), FIV (7.4%) and Ap (1.8%). Bh antibodies were associated with seroreactivity to both Ec and Rc antigens. FIV antibodies were associated with illness and cats older than 2 years. Bartonella henselae and B. clarridgeiae (Bcl) DNA was amplified from seven and one sample, respectively.}, number={3-4}, journal={VETERINARY MICROBIOLOGY}, author={Solano-Gallego, Laia and Hegarty, Barbara and Espada, Yvonne and Llull, Joan and Breitschwerdt, Edward}, year={2006}, month={Dec}, pages={274–277} } @article{gary_webb_hegarty_breitschwerdt_2006, title={The low seroprevalence of tick-transmitted agents of disease in dogs from southern Ontario and Quebec}, volume={47}, number={12}, journal={Canadian Veterinary Journal}, author={Gary, A. T. and Webb, J. A. and Hegarty, B. C. and Breitschwerdt, E. B.}, year={2006}, pages={1194–1200} } @article{lester_breitschwerdt_collis_hegarty_2005, title={Anaplasma phagocytophilum infection (granulocytic anaplasmosis) in a dog from Vancouver Island}, volume={46}, number={9}, journal={Canadian Veterinary Journal}, author={Lester, S. J. and Breitschwerdt, E. B. and Collis, C. D. and Hegarty, B. C.}, year={2005}, pages={825–827} } @article{breitschwerdt_hegarty_maggi_hawkins_dyer_2005, title={Bartonella Species as a Potential Cause of Epistaxis in Dogs}, volume={43}, ISSN={0095-1137}, url={http://dx.doi.org/10.1128/JCM.43.5.2529-2533.2005}, DOI={10.1128/JCM.43.5.2529-2533.2005}, abstractNote={ABSTRACT Infection with a Bartonella species was implicated in three cases of epistaxis in dogs, based upon isolation, serology, or PCR amplification. These cases, in conjunction with previously published reports, support a potential role for Bartonella spp. as a cause of epistaxis in dogs and potentially in other animals, including humans.}, number={5}, journal={Journal of Clinical Microbiology}, publisher={American Society for Microbiology}, author={Breitschwerdt, E. B. and Hegarty, B. C. and Maggi, R. and Hawkins, E. and Dyer, P.}, year={2005}, month={May}, pages={2529–2533} } @article{o'rourke_pitulle_hegarty_kraycirik_killary_grosenstein_brown_breitschwerdt_2005, title={Bartonella quintana in Cynomolgus Monkey (Macaca fascicularis)}, volume={11}, ISSN={1080-6040 1080-6059}, url={http://dx.doi.org/10.3201/eid1112.030045}, DOI={10.3201/eid1112.030045}, abstractNote={We identified a Bartonella quintana strain by polymerase chain reaction amplification, cloning, and sequencing of DNA extracted from lysed erythrocytes and cultured colonies grown from peripheral blood collected from a captive-bred cynomolgus monkey (Macaca fascicularis). This report describes naturally acquired B. quintana infection in a nonhuman primate.}, number={12}, journal={Emerging Infectious Diseases}, publisher={Centers for Disease Control and Prevention (CDC)}, author={O'Rourke, Laurie G. and Pitulle, Christian and Hegarty, Barbara C. and Kraycirik, Sharon and Killary, Karen A. and Grosenstein, Paul and Brown, James W. and Breitschwerdt, Edward B.}, year={2005}, month={Dec}, pages={1931–1934} } @article{solano-gallego_bradley_hegarty_sigmon_breitschwerdt_2004, title={Bartonella henselae IgG antibodies are prevalent in dogs from southeastern USA}, volume={35}, ISSN={["0928-4249"]}, DOI={10.1051/vetres:2004034}, abstractNote={In contrast to the large body of literature regarding Bartonella henselae in humans and cats, there is little information about B. henselae as an infectious agent in dogs. Due to the paucity of information regarding the B. henselae serology in dogs, we performed a cross-sectional serosurvey using B. henselae antigen in order to compare the seroprevalence between sick and healthy dogs from the south-eastern USA. Ninety-nine sera were collected from clinically healthy dogs. Three hundred and one sera from sick dogs were submitted to North Carolina State University for serologic screening against a panel of arthropod-transmitted organisms. Serological tests were performed using B. henselae (Bh), Rickettsia rickettsii (Rr), Ehrlichia canis (Ec), Bartonella vinsonii subspecies berkhoffii (Bvb), Babesia canis (Bc) and Borrelia burgdorferi (Bb) antigens. Serum B. henselae IgG antibodies were detected in 10.1% of healthy dogs and in 27.2% of sick dogs. The difference in seroprevalence between the two groups was statistically significant. The majority of seroreactive dogs (80%) had low titers of 1:64 or 1:128. In healthy dogs, seroprevalence for Rr was 14.1% and for Bvb was 1%. In sick dogs, Rr seroprevalence was 29.7%, Ec 6.5%, Bvb 4.7%, Bb 1.7% and Bc was 0.85%. Of the sick dogs that were seroreactive to B. henselae antigens, 40.6% were also seroreactive to Rr, 15.0% reactive to Bvb antigens, 14.8% reactive to Ec antigens, 1.8% reactive to Bc antigens and 1.75% reactive to Bb antigens. Sera from dogs experimentally infected with B. vinsonii subsp. berkhoffii, E. canis or R. rickettsii did not cross react with B. henselae antigens, by IFA testing. This study indicates that B. henselae IgG antibodies are prevalent in healthy and sick dogs living in the south-eastern USA. Nevertheless, further studies are needed to evaluate the epidemiological, clinical and zoonotic relevance of B. henselae infection in dogs.}, number={5}, journal={VETERINARY RESEARCH}, author={Solano-Gallego, L and Bradley, J and Hegarty, B and Sigmon, B and Breitschwerdt, E}, year={2004}, pages={585–595} } @article{mylonakis_koutinas_breitschwerdt_hegarty_billinis_leontides_kontos_2004, title={Chronic canine ehrlichiosis (Ehrlichia canis): A retrospective study of 19 natural cases}, volume={40}, ISSN={["0587-2871"]}, DOI={10.5326/0400174}, abstractNote={Nineteen dogs from Greece with chronic ehrlichiosis were studied. The dogs exhibited bicytopenia or pancytopenia, bone marrow hypoplasia, seroreactivity to Ehrlichia canis (E. canis) antigens, and had no history of drug or radiation exposure. Anorexia, depression, severe bleeding tendencies, hypoalbuminemia, and increased serum alanine aminotransferase activity were also hallmarks of the disease. All these animals eventually died, irrespective of the treatment applied. Some dogs were also serologically positive for Rickettsia conorii, Leishmania infantum (L. infantum), and Bartonella vinsonii subspp. berkhoffii. Polymerase chain reaction testing of bone marrow samples revealed E. canis, Anaplasma phagocytophilia, Anaplasma platys, and L. infantum in some dogs. Concurrent infections did not appear to substantially influence the clinical course and final outcome of the chronic canine ehrlichiosis.}, number={3}, journal={JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION}, author={Mylonakis, ME and Koutinas, AF and Breitschwerdt, EB and Hegarty, BC and Billinis, CD and Leontides, LS and Kontos, VS}, year={2004}, pages={174–184} } @article{seaman_kania_hegarty_legendre_breitschwerdt_2004, title={Comparison of results for serologic testing and a polymerase chain reaction assay to determine the prevalence of stray dogs in eastern Tennessee seropositive to Ehrlichia canis}, volume={65}, ISSN={0002-9645}, url={http://dx.doi.org/10.2460/ajvr.2004.65.1200}, DOI={10.2460/ajvr.2004.65.1200}, abstractNote={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.}, number={9}, journal={American Journal of Veterinary Research}, publisher={American Veterinary Medical Association (AVMA)}, author={Seaman, Rebecca L. and Kania, Stephen A. and Hegarty, Barbara C. and Legendre, Alfred M. and Breitschwerdt, Edward B.}, year={2004}, month={Sep}, pages={1200–1203} } @article{ober_spaulding_breitschwerdt_malarkey_hegarty_2004, title={ORCHITIS IN TWO DOGS WITH ROCKY MOUNTAIN SPOTTED FEVER}, volume={45}, ISSN={1058-8183 1740-8261}, url={http://dx.doi.org/10.1111/j.1740-8261.2004.04079.x}, DOI={10.1111/j.1740-8261.2004.04079.x}, abstractNote={Two dogs with testicular swelling were sonographically diagnosed with orchitis and were subsequently diagnosed with Rocky Mountain spotted fever (RMSF). Use of both gray scale and color Doppler sonography allowed for differentiation of orchitis from neoplasia and torsion. While only experimentally induced RMSF is reported to cause orchitis in dogs, it should be considered in any dog with vascular insult to the testes, especially when other signs of systemic illness are involved.}, number={5}, journal={Veterinary Radiology & Ultrasound}, publisher={Wiley}, author={Ober, Christopher P. and Spaulding, Kathy and Breitschwerdt, Edward B. and Malarkey, David E. and Hegarty, Barbara C.}, year={2004}, month={Sep}, pages={458–465} } @article{goodman_hawkins_olby_grindem_hegarty_breitschwerdt_2003, title={Molecular identification of Ehrlichia ewingii infection in dogs: 15 cases (1997-2001)}, volume={222}, DOI={10.2460/javma.2003.222.1102}, abstractNote={OBJECTIVE To determine historical, physical examination, hematologic, and serologic findings in dogs with Ehrlichia ewingii infection. DESIGN Retrospective study. ANIMALS 15 dogs. PROCEDURE In all dogs, infection with E ewingii was confirmed with a polymerase chain reaction (PCR) assay. Follow-up information and clarification of information recorded in the medical records was obtained by telephone interviews and facsimile correspondence with referring veterinarians and owners. RESULTS Fever and lameness were the most common findings with each occurring in 8 dogs. Five dogs had neurologic abnormalities including ataxia, paresis, proprioceptive deficits, anisocoria, intention tremor, and head tilt. Neutrophilic polyarthritis was identified in 4 dogs. No clinical signs were reported in 3 dogs. The predominant hematologic abnormality was thrombocytopenia, which was identified in all 12 dogs for which a platelet count was available. Reactive lymphocytes were seen in 5 of 13 dogs. Concurrent infection with another rickettsial organism was identified in 4 dogs. Of the 13 dogs tested, 7 were seroreactive to E canis antigens. Morulae consistent with E ewingii infection were identified in neutrophils in 8 dogs. Treatment with doxycycline, with or without prednisone, resulted in a rapid, favorable clinical response in the 9 dogs for which follow-up information was available. CONCLUSIONS AND CLINICAL RELEVANCE Results suggest that PCR testing for E ewingii infection should be considered in dogs with fever, neutrophilic polyarthritis, unexplained ataxia or paresis, thrombocytopenia, or unexplained reactive lymphocytes, and in dogs with clinical signs suggestive of ehrlichiosis that are seronegative for E canis. Following treatment with doxycycline, the prognosis for recovery is good.}, number={8}, journal={JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION}, author={Goodman, RA and Hawkins, EC and Olby, NJ and Grindem, CB and Hegarty, B and Breitschwerdt, EB}, year={2003}, month={Apr}, pages={1102–1107} } @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{beaufils_breitschwerdt_hancock_hegarty_martin-granel_jumelle_barbault-jumelle_blavier_2002, title={Feline ehrlichiosis. The genetic identification of the agent in Avo cats}, volume={37}, number={3}, journal={Pratique Medicale et Chirurgicale de L'animal de Compagnie}, author={Beaufils, J. P. and Breitschwerdt, E. and Hancock, S. I. and Hegarty, B. C. and Martin-Granel, J. and Jumelle, P. and Barbault-Jumelle, M. and Blavier, A.}, year={2002}, pages={235–238} } @article{breitschwerdt_abrams-ogg_lappin_bienzle_hancock_cowan_clooten_hegarty_hawkins_2002, title={Molecular evidence supporting Ehrlichia canis-like infection in cats}, volume={16}, ISSN={["0891-6640"]}, DOI={10.1892/0891-6640(2002)016<0642:MESCII>2.3.CO;2}, abstractNote={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.}, number={6}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Breitschwerdt, EB and Abrams-Ogg, ACG and Lappin, MR and Bienzle, D and Hancock, SI and Cowan, SM and Clooten, JK and Hegarty, BC and Hawkins, EC}, year={2002}, pages={642–649} } @article{williams_van steenhouse_bradley_hancock_hegarty_breitschwerdt_2002, title={Naturally OccurringEhrlichia chaffeensisInfection in Two Prosimian Primate Species: Ring-tailed Lemurs (Lemur catta) and Ruffed Lemurs (Varecia variegata)}, volume={8}, ISSN={1080-6040 1080-6059}, url={http://dx.doi.org/10.3201/eid0812.020085}, DOI={10.3201/eid0812.020085}, abstractNote={A naturally occurring infection of Ehrlichia chaffeensis in lemurs is described. DNA of Ehrlichia chaffeensis was identified by polymerase chain reaction in peripheral blood from six of eight clinically ill lemurs. Organisms were cultured from the blood of one lemur exhibiting clinical and hematologic abnormalities similar to those of humans infected with E. chaffeensis.}, number={12}, journal={Emerging Infectious Diseases}, publisher={Centers for Disease Control and Prevention (CDC)}, author={Williams, Cathy V. and Van Steenhouse, Jan L. and Bradley, Julie M. and Hancock, Susan I. and Hegarty, Barbara C. and Breitschwerdt, Edward B.}, year={2002}, month={Dec}, pages={1497–1500} } @article{gaskin_schantz_jackson_birkenheuer_tomlinson_gramiccia_levy_steurer_kollmar_hegarty_et al._2002, title={Visceral leishmaniasis in a New York foxhound kennel}, volume={16}, ISSN={["0891-6640"]}, DOI={10.1892/0891-6640(2002)016<0034:VLIANY>2.3.CO;2}, abstractNote={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.}, number={1}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Gaskin, AA and Schantz, P and Jackson, J and Birkenheuer, A and Tomlinson, L and Gramiccia, M and Levy, M and Steurer, F and Kollmar, E and Hegarty, BC and et al.}, year={2002}, pages={34–44} } @article{hinrichsen_whitworth_breitschwerdt_hegarty_mather_2001, title={Assessing the association between the geographic distribution of deer ticks and seropositivity rates to various tick-transmitted disease organisms in dogs}, volume={218}, ISSN={0003-1488}, url={http://dx.doi.org/10.2460/javma.2001.218.1092}, DOI={10.2460/javma.2001.218.1092}, abstractNote={Abstract}, number={7}, journal={Journal of the American Veterinary Medical Association}, publisher={American Veterinary Medical Association (AVMA)}, author={Hinrichsen, Virginia L. and Whitworth, Ulysses G. and Breitschwerdt, Edward B. and Hegarty, Barbara C. and Mather, Thomas N.}, year={2001}, month={Apr}, pages={1092–1097} } @article{munana_vitek_hegarty_kordick_breitschwerdt_2001, title={Infection of Fetal Feline Brain Cells in Culture with Bartonella henselae}, volume={69}, ISSN={0019-9567}, url={http://dx.doi.org/10.1128/IAI.69.1.564-569.2001}, DOI={10.1128/IAI.69.1.564-569.2001}, abstractNote={ABSTRACT}, number={1}, journal={Infection and Immunity}, publisher={American Society for Microbiology}, author={Munana, K. R. and Vitek, S. M. and Hegarty, B. C. and Kordick, D. L. and Breitschwerdt, E. B.}, year={2001}, month={Jan}, pages={564–569} } @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, including Ehrlichia sp. (Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia equi, and Ehrlichia risticii), Bartonella vinsonii subsp. berkhoffi (Bvb), spotted fever group (SFG) rickettsiae (Rickettsia rickettsii), Typhus group (TG) rickettsiae (Rickettsia canada, Rickettsia prowazekii, and Rickettsia typhi), and Babesia sp. (Babesia canis and Babesia 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} } @article{kordick_breitschwerdt_hegarty_southwick_colitz_hancock_bradley_rumbough_mcpherson_maccormack_1999, title={Coinfection with multiple tick-borne pathogens in a Walker Hound kennel in North Carolina}, volume={37}, number={8}, journal={Journal of Clinical Microbiology}, author={Kordick, S. K. and Breitschwerdt, E. B. and Hegarty, B. C. and Southwick, K. L. and Colitz, C. M. and Hancock, S. I. and Bradley, J. M. and Rumbough, R. and McPherson, J. T. and MacCormack, J. N.}, year={1999}, pages={2631–2638} } @article{breitschwerdt_papich_hegarty_gilger_hancock_davidson_1999, title={Efficacy of Doxycycline, Azithromycin, or Trovafloxacin for Treatment of Experimental Rocky Mountain Spotted Fever in Dogs}, volume={43}, ISSN={0066-4804 1098-6596}, url={http://dx.doi.org/10.1128/AAC.43.4.813}, DOI={10.1128/aac.43.4.813}, abstractNote={ABSTRACT}, number={4}, journal={Antimicrobial Agents and Chemotherapy}, publisher={American Society for Microbiology}, author={Breitschwerdt, E. B. and Papich, M. G. and Hegarty, B. C. and Gilger, B. and Hancock, S. I. and Davidson, M. G.}, year={1999}, month={Apr}, pages={813–821} } @article{grindem_breitschwerdt_perkins_cullins_thomas_hegarty_1999, title={Platelet-associated immunoglobulin (antiplatelet antibody) in canine Rocky Mountain spotted fever and ehrlichiosis}, volume={35}, ISSN={["0587-2871"]}, DOI={10.5326/15473317-35-1-56}, abstractNote={Antiplatelet antibodies were detected in the sera of dogs with naturally occurring and experimentally induced Rickettsia rickettsii and Ehrlichia canis infections. This is the first known report documenting elevated platelet-associated immunoglobulin (PAIg) titers in Rocky Mountain spotted fever (RMSF) infections. In the naturally occurring RMSF infections and ehrlichiosis, the antibodies persisted for weeks or months, even when the platelet counts had normalized. Results of this study indicate an immunological component for rickettsial thrombocytopenia. Therefore, current therapeutic recommendations, especially regarding avoiding the use of immunosuppressive drugs in patients with rickettsial diseases, need to be critically reviewed.}, number={1}, journal={JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION}, author={Grindem, CB and Breitschwerdt, EB and Perkins, PC and Cullins, LD and Thomas, TJ and Hegarty, BC}, year={1999}, pages={56–61} } @article{carpenter_gandhi_kong_corey_chen_walker_dumler_breitschwerdt_hegarty_sexton_et al._1999, title={The incidence of ehrlichial and rickettsial infection in patients with unexplained fever and recent history of tick bite in central North Carolina}, volume={180}, ISSN={["0022-1899"]}, DOI={10.1086/314954}, abstractNote={We examined the clinical and laboratory findings of a consecutive series of patients from central North Carolina presenting with fever and a history of tick bite within the preceding 14 days. Evidence of a tick-transmitted pathogen was detected in 16 of 35 patients enrolled over a 2-year period. Nine patients were infected with Ehrlichia chaffeensis, and 6 were infected with a spotted fever group rickettsia; 1 patient had evidence of coinfection with E. chaffeensis and a spotted fever group rickettsia. Four patients had detectable antibodies against the human granulocytic ehrlichiosis agent; however, only 2 had a 4-fold antibody titer rise without detectable antibodies against E. chaffeensis. The other 2 were thought to have cross-reacting antibodies to E. chaffeensis. We conclude that ehrlichial infections may be as common as spotted fever group rickettsial infections in febrile patients from central North Carolina with a recent history of tick bite.}, number={3}, journal={JOURNAL OF INFECTIOUS DISEASES}, author={Carpenter, C. F. and Gandhi, T. K. and Kong, L. K. and Corey, G. R. and Chen, S. M. and Walker, D. H. and Dumler, J. S. and Breitschwerdt, Edward and Hegarty, B. and Sexton, D. J. and et al.}, year={1999}, month={Sep}, pages={900–903} } @article{baneth_breitschwerdt_hegarty_pappalardo_ryan_1998, title={A survey of tick-borne bacteria and protozoa in naturally exposed dogs from Israel}, volume={74}, ISSN={["1873-2550"]}, DOI={10.1016/S0304-4017(97)00149-0}, abstractNote={Antibody reactivity against seven bacterial or protozoal pathogens was measured in sera derived from 40 dogs suspected of a tick-borne disease. Sera from 73% (29/40) of the dogs reacted with three or more test antigens. Seroreactivity was most prevalent to Babesia canis antigen (90%) followed by Babesia gibsoni (75%), Ehrlichia canis (63%), Rickettsia conorii--Moroccan strain (58%), Rickettsia conorii--Israeli strain no. 2 (28%), Borrelia burgdorferi (10%) or Bartonella vinsonii (berkhoffii) (10%). Seroconversion documented in seven dogs, supported an acute phase diagnosis of ehrlichiosis in four dogs, R. conorii infection in three dogs and babesiosis in one dog. In the remaining dogs, correlation of clinical abnormalities with increased seroreactivity was not established through the design of this study. Although Lyme borreliosis has not been reported in people in Israel, Western blot analysis for antibodies reactive to B. burgdorferi identified genus-specific antiflagellin antibodies indicating that dogs in Israel are exposed to a Borrelia species. Identification of species-specific seroreactivity was not possible and infection with a Borrelia species other than B. burgdorferi is likely. Seroreactivity to B. vinsonii (berkhoffii) in dogs outside the USA is reported here for the first time.}, number={2-4}, journal={VETERINARY PARASITOLOGY}, author={Baneth, G and Breitschwerdt, EB and Hegarty, BC and Pappalardo, B and Ryan, J}, year={1998}, month={Jan}, pages={133–142} } @article{breitschwerdt_hegarty_hancock_1998, title={Doxycycline hyclate treatment of experimental canine ehrlichiosis followed by challenge inoculation with two Ehrlichia canis strains}, volume={42}, number={2}, journal={Antimicrobial Agents and Chemotherapy}, author={Breitschwerdt, E. B. and Hegarty, B. C. and Hancock, S. I.}, year={1998}, pages={362–368} } @article{sexton_corey_carpenter_kong_gandhi_breitschwerdt_hegarty_chen_feng_zhao_et al._1998, title={Dual Infection with Ehrlichia chaffeensis and a Spotted Fever Group Rickettsia: A Case Report}, volume={4}, ISSN={1080-6040}, url={http://dx.doi.org/10.3201/eid0402.980222}, DOI={10.3201/eid0402.980222}, abstractNote={A 44-year-old man with a history ofhepatitis C infection and regular use of cocainewas asymptomatic until June 6, 1995, when henoted the acute onset of myalgia, headache,arthralgia, fever (40°C), nonproductive cough,nausea, and vomiting. The following day hesought medical care and received oraltrimethoprim/sulfamethoxazole for a presumedrespiratory tract infection. Over the next 3 days,his symptoms worsened; on the fourth day henoticed a maculopapular rash and came to ourmedical center. Initial examination showedtachycardia (pulse 125 beats per minute), fever(38.6°C), and a macular red rash localized to hisright arm. The patient reported a tick bite 1 daybefore his symptoms began. In addition, hereported removing an engorged tick from his petdog and crushing it between his fingers 4 daysbefore he became ill. Laboratory studies showedleukopenia (white blood cell count 2,700/mm}, number={2}, journal={Emerging Infectious Diseases}, publisher={Centers for Disease Control and Prevention (CDC)}, author={Sexton, Daniel and Corey, G. Ralph and Carpenter, Christopher and Kong, Li Quo and Gandhi, Tejel and Breitschwerdt, Edward B. and Hegarty, Barbara C. and Chen, Sheng-Ming and Feng, Hui-Min and Zhao, Li and et al.}, year={1998}, month={Jun}, pages={311–316} } @article{goldman_breitschwerdt_grindem_hegarty_walls_dumler_1998, title={Granulocytic Ehrlichiosis in Dogs from North Carolina and Virginia}, volume={12}, ISSN={0891-6640 1939-1676}, url={http://dx.doi.org/10.1111/j.1939-1676.1998.tb02096.x}, DOI={10.1111/j.1939-1676.1998.tb02096.x}, abstractNote={Medical records of 3 dogs from North Carolina and 3 dogs from Virginia with ehrlichial morulae in circulating neutrophils were studied retrospectively. Two clinically distinct disease syndromes, including chronic, moderate to severe anemia (n = 3) and polyarthritis (n = 2) were associated with canine granulocytic ehrlichiosis (CGE) in these dogs. One dog was clinically healthy, and abnormalities were not detected during physical examination. Clinical signs were nonspecific and included fever, lethargy, anorexia, vomiting, and diarrhea. The most frequent laboratory abnormalities were normocytic normochromic nonregenerative anemia, moderate thrombocytopenia with large platelets, lymphopenia, and eosinopenia. Considerable variability was found in the serologic responses toEhrlichia equi, Ehrlichia canis, andEhrlichia chaffeensisantigens among the 5 dogs for which stored sera were available for indirect fluorescent antibody testing. Polymerase chain reaction amplification and sequencing of portions of the 16S rRNA gene from blood (collected in ethylenediaminetetraacetic acid) of 1 severely anemic dog (dog 3) and 1 polyarthritic dog (dog 4) resulted in DNA sequences nearly identical to the GenBank accessions forEhrlichia ewingii.The DNA sequence from a 3rd dog (dog 5) was most similar to that ofE. canis.Serologic or molecular results support the possibility ofE. ewingii, E. equi, andE. caniscoinfection or serologic cross‐reactivity among canine granulocytic and monocyticEhrlichiaspecies in dogs from North Carolina and Virginia. Variability in response to tetracycline or doxycycline treatment was noted in these dogs, with more rapid resolution of signs in dogs with polyarthritis. We report the 1st cases of CGE in dogs from North Carolina and Virginia, including recognition of CGE in a healthy dog.}, number={2}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Goldman, Elizabeth E. and Breitschwerdt, Edward B. and Grindem, Carol B. and Hegarty, Barbara C. and Walls, Jennifer J. and Dumler, J. Stephen}, year={1998}, month={Mar}, pages={61–70} } @article{breitschwerdt_hegarty_hancock_1998, title={Sequential evaluation of dogs naturally infected with Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia equi, Ehrlichia ewingii, or Bartonella vinsonii}, volume={36}, number={9}, journal={Journal of Clinical Microbiology}, author={Breitschwerdt, E. B. and Hegarty, B. C. and Hancock, S. I.}, year={1998}, pages={2645–2651} } @article{hegarty_levy_gager_breitschwerdt_1997, title={Immunoblot Analysis of the Immunoglobulin G Response to Ehrlichia Canis in Dogs: An International Survey}, volume={9}, ISSN={1040-6387 1943-4936}, url={http://dx.doi.org/10.1177/104063879700900106}, DOI={10.1177/104063879700900106}, abstractNote={ Historically, considerable variation has been reported in the type and severity of clinical and hematologic abnormalities associated with canine ehrlichiosis. Because of difficulties associated with the isolation of intracellular monocytic Ehrlichia species in tissue culture systems, few E. canis isolates are available for comparative microbiologic studies. To address the issue of potential E. canis antigenic diversity in different regions of the world, dog sera reactive by indirect fluorescent antibody testing to E. canis (Florida) antigen were obtained from France, Israel, Italy, the United States, the Virgin Islands, and Zimbabwe. Ehrlichia canis proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and at least 5 sera from each region were stained by western immunoblotting. Antibody immunodominance was scored based upon staining intensity. There was relative homogeneity in the immunogenic protein reactions to E. canis antigens. Of the 58 E. canis reactive sera, 54 samples resulted in immunoblot patterns indicative of chronic ehrlichiosis. Four reactive sera (reciprocal titers of 160–2,560) did not recognize any genus-specific antigens resulting in protein bands between 22 and 29 kD, indicating serologic cross-reactivity with other microorganisms. Relatively homogenous immunoblot patterns, consistent with the reported immunoblot response of dogs with experimental chronic ehrlichiosis, were observed with sera from Arizona, France, Israel, North Carolina, Texas, and the Virgin Islands. In contrast, unique major proteins were observed in dog sera from Italy and Zimbabwe. Our results indicate that although relatively homogeneous, antigenic diversity may exist among E. canis organisms in different regions of the world. }, number={1}, journal={Journal of Veterinary Diagnostic Investigation}, publisher={SAGE Publications}, author={Hegarty, Barbara C. and Levy, Michael G. and Gager, Robin F. and Breitschwerdt, Edward B.}, year={1997}, month={Jan}, pages={32–38} } @article{breitschwerdt_davidson_hegarty_papich_grindem_1997, title={Prednisolone at anti-inflammatory or immunosuppressive dosages in conjunction with doxycycline does not potentiate the severity of Rickettsia rickettsii infection in dogs.}, volume={41}, ISSN={0066-4804 1098-6596}, url={http://dx.doi.org/10.1128/AAC.41.1.141}, DOI={10.1128/aac.41.1.141}, abstractNote={Dogs were experimentally inoculated with Rickettsia rickettsii to determine if anti-inflammatory or immunosuppressive dosages of prednisolone, when administered in conjunction with an antirickettsial antibiotic (doxycycline), induced therapeutically relevant pathophysiological consequences that ultimately influence disease outcome. Although the duration of rickettsemia was prolonged in dogs receiving immunosuppressive, but not anti-inflammatory, corticosteroids, concurrent administration of doxycycline and corticosteroids conferred no other detected detrimental effects. Treatment with doxycycline or doxycycline in conjunction with prednisolone resulted in decreased R. rickettsii-specific antibody titers; however, examination of appropriately timed acute- and convalescent-phase serum samples would have facilitated an accurate diagnosis of Rocky Mountain spotted fever (RMSF) in all 16 dogs. We conclude that the concurrent use of anti-inflammatory or immunosuppressive doses of prednisolone in conjunction with doxycycline, early in the course of experimental RMSF, confers no clinically relevant detrimental effects and that additional studies might be indicated to detect possible beneficial effects in cases of severe or potentially fulminant RMSF. However, because the illness induced in these dogs was of mild to moderate severity, the results of this study should definitely not be construed as supporting the safety or efficacy of prednisolone for treatment of severe canine or human RMSF.}, number={1}, journal={Antimicrobial Agents and Chemotherapy}, publisher={American Society for Microbiology}, author={Breitschwerdt, E B and Davidson, M G and Hegarty, B C and Papich, M G and Grindem, C B}, year={1997}, month={Jan}, pages={141–147} } @article{breitschwerdt_geoly_meuten_levine_howard_hegarty_stafford_1996, title={Myocarditis in mice and guinea pigs experimentally infected with a canine-origin Borrelia isolate from Florida}, volume={57}, number={4}, journal={American Journal of Veterinary Research}, author={Breitschwerdt, E. B. and Geoly, F. J. and Meuten, D. J. and Levine, J. F. and Howard, P. and Hegarty, B. C. and Stafford, L. C.}, year={1996}, pages={505–511} }