@article{eberts_diniz_beall_stillman_chandrashekar_breitschwerdt_2011, title={Typical and Atypical Manifestations of Anaplasma phagocytophilum Infection in Dogs}, volume={47}, ISSN={["1547-3317"]}, DOI={10.5326/jaaha-ms-5578}, abstractNote={Eighteen clinically ill dogs, naturally infected with Anaplasma phagocytophilum, were examined at a veterinary practice in Baxter, Minnesota. A clinical examination, complete blood cell count, enzyme- linked immunosorbent assay (ELISA) for A phagocytophilum, Borrelia burgdorferi, and Ehrlichia canis antibodies and Dirofilaria immitis antigen, and a polymerase chain reaction test for A phagocytophilum DNA were obtained for all dogs. Physical examination findings included fever, arthropathy, lymphadenopathy, epistaxis, acute gastritis, cervical hyperpathia, and central nervous system dysfunction. Complete blood cell count abnormalities included thrombocytopenia, morulae in neutrophils, anemia, leukopenia, eosinopenia, lymphopenia, and monocytosis. Seroreactivity to A phagocytophilum was found in 61%, B burgdorferi antibodies in 17%, and D immitis antigen in 5% of the dogs. Fever, arthropathy, neurologic dysfunction, and epistaxis are clinical syndromes that can be associated with A phagocytophilum infection. Treatment with doxycycline resulted in rapid resolution of clinical signs in all dogs.}, number={6}, journal={JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION}, author={Eberts, Matthew D. and Diniz, Pedro Paulo Vissotto de Paiva and Beall, Melissa J. and Stillman, Brett A. and Chandrashekar, Ramaswamy and Breitschwerdt, Edward B.}, year={2011}, pages={E86–E94} }
 @article{o'connor_saucier_daniluk_stillman_krah_rikihisa_xiong_yabsley_adams_diniz_et al._2010, title={Evaluation of peptide- and recombinant protein-based assays for detection of anti-Ehrlichia ewingii antibodies in experimentally and naturally infected dogs}, volume={71}, ISSN={0002-9645}, url={http://dx.doi.org/10.2460/ajvr.71.10.1195}, DOI={10.2460/ajvr.71.10.1195}, abstractNote={Abstract}, number={10}, journal={American Journal of Veterinary Research}, publisher={American Veterinary Medical Association (AVMA)}, author={O'Connor, Thomas P. and Saucier, Jill M. and Daniluk, Daryn and Stillman, Brett A. and Krah, Regis and Rikihisa, Yasuko and Xiong, Qingming and Yabsley, Michael J. and Adams, Dustin S. and Diniz, Pedro Paulo V P and et al.}, year={2010}, month={Oct}, pages={1195–1200} }
 @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{perez_hummel_keene_maggi_diniz_breitschwerdt_2010, title={Successful treatment ofBartonella henselaeendocarditis in a cat}, volume={12}, ISSN={1098-612X 1532-2750}, url={http://dx.doi.org/10.1016/j.jfms.2009.12.018}, DOI={10.1016/j.jfms.2009.12.018}, abstractNote={This report describes the clinical presentation, diagnosis and treatment of a cat with vegetative valvular endocarditis temporally associated with natural infection with Bartonella henselae. Lethargy, abnormal gait and weakness were the main clinical signs that resulted in referral for diagnostic evaluation. Using a novel and sensitive culture approach, B henselae was isolated from the blood. Following antibiotic therapy there was total resolution of clinical signs, the heart murmur, the valvular lesion by echocardiography, and no Bartonella species was isolated or amplified from a post-treatment blood culture. In conjunction with previous case reports, infective endocarditis can be associated with natural B henselae infection in cats; however, early diagnosis and treatment may result in a better prognosis than previously reported.}, number={6}, journal={Journal of Feline Medicine and Surgery}, publisher={SAGE Publications}, author={Perez, Cristina and Hummel, James B. and Keene, Bruce W. and Maggi, Ricardo G. and Diniz, Pedro P.V.P. and Breitschwerdt, Edward B.}, year={2010}, month={Jun}, pages={483–486} }
 @article{breitschwerdt_maggi_cadenas_diniz_2009, title={A groundhog, a novel bartonella sequence, and my father's death}, volume={15}, number={12}, journal={Emerging Infectious Diseases}, author={Breitschwerdt, E. B. and Maggi, R. G. and Cadenas, M. B. and Diniz, P. P. V. D.}, year={2009}, pages={2080–2086} }
 @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/μL) 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{diniz_wood_maggi_sontakke_stepnik_breitschwerdt_2009, title={Co-isolation of Bartonella henselae and Bartonella vinsonii subsp. berkhoffii from blood, joint and subcutaneous seroma fluids from two naturally infected dogs}, volume={138}, ISSN={0378-1135}, url={http://dx.doi.org/10.1016/j.vetmic.2009.01.038}, DOI={10.1016/j.vetmic.2009.01.038}, abstractNote={This report describes the clinical presentation, isolation and treatment of two dogs naturally infected with Bartonella henselae and Bartonella vinsonii subsp. berkhoffii. Chronic and progressive polyarthritis was the primary complaint for dog #1, from which B. henselae and B. vinsonii subsp. berkhoffii were cultured on three independent occasions from blood and joint fluid samples, despite administration of nearly 4 months of non-consecutive antibiotic therapy. A clinically atypical and progressively severe trauma-associated seroma was the primary complaint for dog #2, from which B. henselae and B. vinsonii subsp. berkhoffii were isolated from serum, blood and seroma fluid. Dogs can be co-infected with two Bartonella spp. and infection with these organisms should not be ruled out if specific antibodies are not detected. Specialized culture techniques should be used for isolation and to assess antibiotic efficacy.}, number={3-4}, journal={Veterinary Microbiology}, publisher={Elsevier BV}, author={Diniz, Pedro Paulo Vissotto de Paiva and Wood, Michael and Maggi, Ricardo G. and Sontakke, Sushama and Stepnik, Matt and Breitschwerdt, Edward B.}, year={2009}, month={Sep}, pages={368–372} }
 @article{cockwill_taylor_snead_dickinson_cosford_malek_lindsay_diniz_2009, title={Granulocytic anaplasmosis in three dogs from Saskatoon, Saskatchewan}, volume={50}, number={8}, journal={Canadian Veterinary Journal}, author={Cockwill, K. R. and Taylor, S. M. and Snead, E. C. R. and Dickinson, R. and Cosford, K. and Malek, S. and Lindsay, L. R. and Diniz, P. P. V. D.}, year={2009}, pages={835–840} }
 @article{billeter_diniz_battisti_munderloh_breitschwerdt_levy_2009, title={Infection and replication of Bartonella species within a tick cell line}, volume={49}, ISSN={0168-8162 1572-9702}, url={http://dx.doi.org/10.1007/s10493-009-9255-1}, DOI={10.1007/s10493-009-9255-1}, abstractNote={Bartonella species are fastidious, gram negative bacteria, some of which are transmitted by arthropod vectors, including fleas, sandflies, and lice. There is very little information regarding the interaction and/or transmission capabilities of Bartonella species by ticks. In the present study, we demonstrate successful infection of the Amblyomma americanum cell line, AAE12, by seven Bartonella isolates and three Candidatus Bartonella species by electron or light microscopy. With the exception of Bartonella bovis, infection with all other examined Bartonella species induced cytopathic effects characterized by heavy cellular vacuolization and eventually cell lysis. Furthermore, using quantitative real time PCR (qPCR), we demonstrated significant amplification of two B. henselae genotype I isolates in the A. americanum cell line over a 5 days period. Ultimately, tick-cell derived Bartonella antigens may prove useful for the development of more sensitive diagnostic reagents and may assist in the development of an effective vaccine to prevent the further spread of disease caused by these organisms.}, number={3}, journal={Experimental and Applied Acarology}, publisher={Springer Science and Business Media LLC}, author={Billeter, Sarah A. and Diniz, Pedro Paulo V. P. and Battisti, James M. and Munderloh, Ulrike G. and Breitschwerdt, Edward B. and Levy, Michael G.}, year={2009}, month={Feb}, pages={193–208} }
 @article{cherry_diniz_maggi_hummel_hardie_behrend_rozanski_defrancesco_cadenas_breitschwerdt_et al._2009, title={Isolation or Molecular Detection of Bartonella  henselae and Bartonella vinsonii subsp. berkhoffii from Dogs with Idiopathic Cavitary Effusions}, volume={23}, ISSN={0891-6640 1939-1676}, url={http://dx.doi.org/10.1111/j.1939-1676.2008.0246.x}, DOI={10.1111/j.1939-1676.2008.0246.x}, abstractNote={There are a substantial number of pathophysiologic causes for effusions in dogs.1 Previously, using an insect cell culture medium (Bartonella alpha-proteobacteria growth medium—BAPGM), our laboratory isolated Mycobacterium kansasii from a dog with therapeutically intractable pleural effusion.2 Recently, we developed a combined assay incorporating BAPGM pre-enrichment culture, so as to increase Bartonella bacterial numbers, followed by PCR amplification of organism-specific DNA sequences.3 The combined assay has substantially increased the sensitivity of molecular detection of Bartonella infection.3-5 This pre-enrichment culture medium has also isolated numerous other bacterial species of undetermined pathogenicity from human patients in our laboratory.4 In the past decade, several Bartonella species, including B. henselae, B. vinsonii subsp. berkhoffii, B. clarridgeiae, B. elizabethae, B. washoensis, and B. quintana, have been found to infect dogs.6 Currently, case-based evidence suggests that occult infection with these bacteria may contribute to a wide range of disease manifestations in dogs, humans, and potentially other animals.6B. henselae and B. vinsonii subsp. berkhoffii have been reported in association with granulomatous lymphadenitis, hepatic disease,7 and endocarditis in dogs.8 Between July 2004 and December 2007, blood and effusion samples from 20 dogs of varying breeds and ages, were submitted for BAPGM culture by veterinarians at Auburn University, North Carolina State University and Tufts University's Veterinary Teaching Hospitals, of which 5 were found to be infected with a Bartonella spp. Using a previously described approach, DNA was extracted directly from EDTA-anticoagulated blood or thoracic and abdominal effusion samples, from BAPGM pre-enrichment liquid blood and effusion cultures, and from pooled bacterial subculture isolates.3-5, 9 Extracted DNA was screened by PCR with Bartonella genus primers, targeting the 16S-23S intergenic spacer region (ITS), which has a detection limit of 2–3 genomic copies per reaction.9 ITS amplicons were sequenced to confirm the Bartonella species and strain.5, 9 This study provides the first molecular and microbiologic evidence to support infection with B. henselae and B. vinsonii subsp. berkhoffii in dogs with idiopathic cavitary effusions or constrictive pericarditis. An 8-year-old-female spayed Labrador mix was referred to Tufts for evaluation of recurrent pleural effusion. Thoracic fluid cytology was consistent with a transudate with a specific gravity of 1.019, total protein of 2.6 g/dL and 255 cells/μL. No bacterial growth or neoplastic cells were documented. A CBC, serum biochemical profile, coagulation profile, and urinalysis were unremarkable except for mild hyperbilirubinemia and hematuria. A torsed lung lobe was surgically removed. Postoperatively despite administration of azathioprine, prednisone, and spironolactone, thoracocentesis was required for 10 consecutive months. When immunosuppressive therapy was tapered, pleural effusion rapidly recurred. Antibiotics, including doxycycline, azithromycin, or enrofloxacin, did not decrease the rate of fluid accumulation, which had become a modified transudate. The dog underwent an exploratory thoracotomy and died postoperatively. B. henselae strain Houston-I was isolated and sequenced from the thoracic fluid and B. henselae strain Houston-I and B. vinsonii subsp. berkhoffii genotype II were isolated from a BAPGM blood culture. Bartonella serology was not requested. A 3-year-old-female spayed Leonberger was referred to Auburn for evaluation of lethargy, anorexia, weight loss (11 kg), and cervical pain. The only central nervous system abnormality was mild pain on cervical flexion. A possible abdominal mass was noted. There were no hematologic abnormalities. Serum biochemical abnormalities included increased alkaline phosphatase (130 U/L; reference range, 4–95) and alanine amino transferase activities (385 U/L; reference range, 26–200), increased serum creatinine concentration (1.5 mg/dL; reference range, 0.68–1.45), and hypoglobulinemia (2.1 g/dL; reference range, 2.6–5.0). Urinalysis was within normal limits and urine culture failed to grow bacteria. Thoracic radiographs indicated moderate pleural effusion. Ultrasonographic abnormalities included a thickened, stiff stomach wall, abdominal effusion, and irregular kidney margins. An echocardiogram identified no cardiac abnormalities. Pleural fluid analysis was consistent with a modified transudate with a specific gravity of 1.026, total protein of 3.9 g/dL, and 670 total cells/μL. Serum antibodies were not detected to Neospora caninum, Ehrlichia canis, Rickettsia rickettsii, Toxoplasma gondii, canine distemper virus, and Pythium insidiosum. Abdominal exploratory surgery identified a large volume of grossly pink peritoneal fluid. No histologic lesions were found in stomach, liver, omentum, duodenum, mesenteric, and pyloric lymph nodes, kidney, pancreas, and jejunum. B. henselae and B. vinsonii subsp. berkhoffii antibodies were not detected, but B. vinsonii subsp. berkhoffii genotype II was sequenced from the BAPGM pre-enrichment blood culture and the subculture isolate. Azithromycin 250 mg daily and doxycycline 100 mg PO q12 h were administered for 14 days. Nine days after beginning treatment, the dog was eating and drinking normally but pleural effusion failed to resolve. Antibiotics were continued for 2 more weeks. During the next year, the cervical pain continued. The dog was euthanized because of severe peripheral edema with spontaneous fluid extravasation 2 years later. An insulin-dependent diabetic 12-year-old-male castrated Vizsla was evaluated for coughing, gagging, and lethargy. Idiopathic chylothorax initially was diagnosed by the referring veterinarian based on cytological evaluation, which was confirmed by pleural fluid analysis at NCSU-VTH as chylous with a specific gravity of 1.023, total protein of 3.3 g/dL, and 1,940 total cells/μL. A CBC was normal. Biochemical abnormalities included hyperglycemia (serum glucose concentration 258 g/dL; reference range, 73–116), and increased SAP (270 U/L; reference range, 15–156), and ALT activities (100 U/L; reference range, 16–73). Abdominal and thoracic ultrasonography, an echocardiogram, and a helical computed tomography (CT) scan of the thorax and anterior abdomen with lymphangiography were unremarkable, except for pleural effusion. Thoracic duct ligation and partial pericardectomy were performed. The dog recovered uneventfully but continued to accumulate lesser quantities of thoracic fluid, consistently characterized as a modified transudate. Two months postsurgery, the dog developed lethargy, inability to rise, and edema involving the neck, sternum, and right front leg. Urine and pleural fluid cultures failed to grow bacteria. Pleural fluid again was classified as a modified transudate, containing atypical mesothelial cells. A seroma, accompanied by spontaneous leakage of fluid, formed in the ventral neck region. The owner elected euthanasia before availability of the BAPGM culture results. B. henselae DNA was amplified and sequenced directly from the pleural fluid and from the BAPGM enrichment culture. After subculture, an Arthrobacter spp. was isolated, as defined by sequencing the 16S rRNA gene. Serology was not requested. At necropsy, no cause was identified for the severe emaciation, pleural effusion, and subcutaneous edema. A 5-year-old-female spayed Pomeranian was referred to the NCSU-VTH for diagnostic evaluation of panhypoproteinemia. The owners reported infrequent bouts of diarrhea and poorly localized abdominal pain. Serum total protein concentration (2.6 g/dL; reference range, 5.1–7.4), albumin (1.3 g/dL; reference range, 2.8–4.0), and globulin concentrations (1.3 g/dL; reference range, 2.0–4.1) were low. Hematologic abnormalities included lymphopenia (283 lymphocytes/μL; reference range, 480–3,762) and neutrophilia (13,745 neutrophils/μL; reference range, 2,529–12,876). With the exception of bilirubinuria, urinalysis was unremarkable, urine protein/creatinine ratio was normal, and urine culture was negative. Radiographs confirmed thoracic and abdominal effusion. Ultrasonography identified bicavitary anechoic effusion, consistent with a transudate, a collapsed right lung lobe, and portal vein thrombosis. The liver was hypoechoic, abdominal lymph nodes slightly enlarged, and the intestinal lumen was distended with fluid. Echocardiography did not identify pericardial effusion or support a diagnosis of right-sided heart failure. By abdominocentesis, a clear transudative fluid with a specific gravity of 1.005 was obtained. Aerobic and anaerobic culture of the abdominal fluid failed to grow bacteria. Endoscopic gastric biopsies were unremarkable, whereas the duodenum was moderately infiltrated with lymphocytes and plasma cells, accompanied by multifocal lymphangiectasia. A heartworm antigen test was negative. Antibodies to B. henselae and B. vinsonii subsp. berkhoffii, R. rickettsii, E. canis, and Babesia canis were not detected by IFA testing. B. vinsonii subsp. berkhoffii genotype II was isolated from pleural fluid and B. henselae from the blood. Protein-losing enteropathy, accompanied by a portal vein thrombosis, was diagnosed. Treatment consisted of a high protein, low residue diet and metronidazole 10.5 mg/kg PO q8h for 4 weeks. Within 2 weeks, the portal vein thrombosis was no longer visible by ultrasonography. Serum protein concentrations increased progressively and were within reference ranges (total protein, 5.8 g/dL; albumin, 3.6 g/dL; globulin, 2.2 g/dL) by 3 months after initial evaluation. Effusion resolved and did not recur during a 3-year follow-up period. A 6-year-old-female spayed Labrador Retriever was referred for diagnostic evaluation of abdominal effusion. Historically, the dog had remained alert and active. Six liters of serosanguineous, modified transudate with a specific gravity of 1.026, total protein of 4 g/dL and 770 total cells/μL was aspirated from the abdomen. There were no hematologic or coagulation abnormalities, except hypocholesterolemia (134 mg/dL; reference range, 138–317). Central venous pressure was slightly increased (8–9 mmHg). Abdominal and thoracic CT scans identified distention of the hepatic veins and the cranial and caudal vena cava, indicative of right-sided heart failure. By echocardiography, minimal pericardial effusion and hypokinetic left ventricle free wall were documented. Catheterization of the right side of the heart detected a cranial vena cava pressure of 16.1 mmHg, right atrial pressure of 17.4 mmHg, right ventricle pressure at the end of the diastole of 15.6 mmHg and wedge pressure of 17.4 mmHg. The tracing of the right atrial pressure, with prominent x and y descents, was consistent with constrictive pericarditis. Subtotal pericardectomy was performed. On histopathology, the pericardium was variably thickened (up to 0.5 cm) with densely packed collagen fibers, containing few blood vessels and rare hemosiderin-laden macrophages. No etiologic agents were seen, no fungal species were isolated and aerobic and anaerobic blood cultures were negative. On day 8 postpericardectomy, the dog became febrile (T 104°F), vomited, and had diarrhea. A CBC was unremarkable. Two liters of serosanguineous fluid, removed by thoracocentesis, was classified cytologically as an inflammatory sterile effusion containing neutrophils, some phagocytosed by macrophages, and large, atypical mesothelial cells. Anaerobic and aerobic cultures again were negative, potentially because amoxicillin-clavulanate was started by the referring veterinarian 36 hours before sample collection. The treatment regimen was changed to ciprofloxacin, azithromycin, and metoclopramide. Pleural fluid, cultured in BAPGM, resulted in the isolation of Bartonella spp., which was not successfully sequenced. Serum was not available for Bartonella IFA testing. Once culture results became available, treatment with azithromycin (7.8 mg/kg PO q2h) was begun for 5 weeks. At reevaluation 1 month later, the dog was eating normally, active, and alert, and there was no radiographic evidence of thoracic or abdominal effusion. Nine months postpericardectomy, the dog remained healthy with normal exercise tolerance when running and swimming. In this study, we detected infection with B. henselae, B. vinsonii subsp. berkhoffii or both organisms in 5 dogs ranging in age from 3 to 12 years that were diagnosed with pleural, pericardial (constrictive pericarditis), or abdominal effusion. Clinical signs generally were nonspecific and included lethargy, fever, vomiting, diarrhea, abdominal distention, lameness, and cervical pain. DNA was not amplified directly from 3 of 3 blood samples or from 2 of 3 effusion samples, but was amplified from pre-enrichment BAPGM blood and effusion cultures and from subculture isolates, a finding that further supports the utility of the enrichment process before PCR for more optimal detection of infection with a Bartonella spp.3, 5, 9 In humans, thoracic effusions have been reported as infrequent sequelae of Cat Scratch Disease, caused by B. henselae and due to infection with B. quintana.10 Unfortunately, BAPGM cultures often were established after administration of antibiotics. If blood and effusion samples had been cultured earlier in the course of illness and before antibiotic therapy, enhanced detection of Bartonella or other fastidious bacteria may have been achieved in other cases. As extravascular accumulation of fluid is always caused by a pathologic process and as dogs can be chronically infected with B. vinsonii subsp. berkhoffii for at least a year,11 microbiologic and molecular documentation of infection with this genus of bacteria may reflect an opportunistic role, a cofactor in disease expression or a primary pathogenic role in various patients with cavitary effusion. Exudates are the expected inflammatory response induced by bacterial infection, but this paradigm may not apply to Bartonella spp. Recently, the lipopolysaccharide of B. quintana was shown to have anti-inflammatory rather than proinflammatory properties.12 In addition, previous experimental infection studies in dogs suggest that B. vinsonii subsp. berkhoffii is associated with organism-induced immunosuppression.11 These and other unknown factors potentially could contribute to the development of an effusion in the absence of a strong host inflammatory response. Recent experimental studies using rodent models emphasize the ability of Bartonella spp. to invade vascular endothelial cells.13 In immunocompromised people, endothelial infection with B. henselae induces single or multiple vasoproliferative lesions (peliosis hepatis and bacillary angiomatosis).14, 15 Therefore, it is plausible that vascular endothelial infection contributes to increased vascular permeability and aberrant fluid accumulation. Despite isolation or molecular detection of B. henselae and B. vinsonii subsp. berkhoffii in these dogs, no direct cause and effect association can be implicated. However, our results may be clinically relevant because most idiopathic effusions obtained from dogs generally are considered aseptic based on conventional microbiological culture approaches. Two dogs in this study, for which serum was available for testing, were not seroreactive to B. henselae and B. vinsonii subsp. berkhoffii antigens by IFA testing, as described previously.9 Antigenic variability among B. henselae test strains previously has resulted in false negative B. henselae IFA results in human patients with suspected cat scratch disease,16 and a similar occurrence may explain discrepant serology and PCR results. The concept that bacterial infection in transudates or modified transudates obtained from dogs is an infrequent occurrence should be reassessed in the context of Bartonella infection. This research was supported by the State of North Carolina and in part through graduate student stipend support provided to NA Cherry by Novartis Animal Health, and salary support provided by IDEXX Laboratories and Bayer Corporation. We thank Mrs Tonya Lee for editorial assistance.}, number={1}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Cherry, N. A. and Diniz, P. P. V. P. and Maggi, Ricardo and Hummel, J. B. and Hardie, E. M. and Behrend, E. N. and Rozanski, E. and DeFrancesco, T. C. and Cadenas, M. B. and Breitschwerdt, Edward and et al.}, year={2009}, month={Jan}, pages={186–189} }
 @article{diniz_billeter_otranto_de caprariis_petanides_mylonakis_koutinas_breitschwerdt_2009, title={Molecular Documentation of Bartonella Infection in Dogs in Greece and Italy}, volume={47}, ISSN={0095-1137}, url={http://dx.doi.org/10.1128/JCM.00082-09}, DOI={10.1128/JCM.00082-09}, abstractNote={ABSTRACT}, number={5}, journal={Journal of Clinical Microbiology}, publisher={American Society for Microbiology}, author={Diniz, P. P. V. P. and Billeter, S. A. and Otranto, D. and De Caprariis, D. and Petanides, T. and Mylonakis, M. E. and Koutinas, A. F. and Breitschwerdt, E. B.}, year={2009}, month={Mar}, pages={1565–1567} }
 @article{sontakke_cadenas_maggi_diniz_breitschwerdt_2009, title={Use of broad range16S rDNA PCR in clinical microbiology}, volume={76}, ISSN={0167-7012}, url={http://dx.doi.org/10.1016/j.mimet.2008.11.002}, DOI={10.1016/j.mimet.2008.11.002}, abstractNote={Broad range 16S rDNA PCR can be used to facilitate the diagnosis of infectious diseases of bacterial origin by detecting 16S rDNA sequences in patient samples. Post amplification sequencing facilitates identification of the infecting organism, but may not allow for differentiation at the species or strain level. This review focuses on the historical use and current applications of broad range 16S rDNA PCR in the diagnosis of bacterial infection. Use of an enrichment liquid culture prior to PCR and the use of real time PCR are also considered. A review of the literature indicates that the diagnostic utility of broad range 16S rDNA PCR is enhanced substantially, if the detected organism is a well-documented pathogen. Frequent detection of environmental organisms of undetermined pathogenicity is currently a limitation. This review also examines weighted criteria developed by different researchers and proposes a decision making tree that establishes the relative importance of various criteria for attributing diagnostic relevance when evaluating individual patient samples. Based upon our review of the literature, a more uniform consensus on the accurate interpretation of broad range 16S rDNA PCR results are needed to improve the microbiological utility of this modality for the diagnosis of bacterial infections in animals and in human patients.}, number={3}, journal={Journal of Microbiological Methods}, publisher={Elsevier BV}, author={Sontakke, Sushama and Cadenas, Maria B. and Maggi, Ricardo G. and Diniz, Pedro Paulo V.P. and Breitschwerdt, Edward B.}, year={2009}, month={Mar}, pages={217–225} }
 @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{petanides_koutinas_mylonakis_day_saridomichelakis_leontides_mischke_diniz_breitschwerdt_kritsepi_et al._2008, title={Factors Associated with the Occurrence of Epistaxis in Natural Canine Leishmaniasis (Leishmania infantum)}, volume={22}, ISSN={0891-6640 1939-1676}, url={http://dx.doi.org/10.1111/j.1939-1676.2008.0129.x}, DOI={10.1111/j.1939-1676.2008.0129.x}, abstractNote={Background:  Canine leishmaniasis (CanL) is a common cause of epistaxis in dogs residing in endemic areas. The pathogenesis of CanL‐associated epistaxis has not been fully explored because of the limited number of cases reported so far.}, number={4}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Petanides, T.A. and Koutinas, A.F. and Mylonakis, M.E. and Day, M.J. and Saridomichelakis, M.N. and Leontides, L.S. and Mischke, R. and Diniz, P. and Breitschwerdt, E.B. and Kritsepi, M. and et al.}, year={2008}, month={Jul}, pages={866–872} }
 @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{diniz_de morais_breitschwerdt_schwartz_2008, title={Serum Cardiac Troponin I Concentration in Dogs with Ehrlichiosis}, volume={22}, ISSN={0891-6640 1939-1676}, url={http://dx.doi.org/10.1111/j.1939-1676.2008.0145.x}, DOI={10.1111/j.1939-1676.2008.0145.x}, abstractNote={Background:  Ehrlichiosis is a multisystemic disease with the potential to cause cardiomyocyte injury in naturally infected dogs.}, number={5}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Diniz, P.P.V.P. and de Morais, H.S.A. and Breitschwerdt, E.B. and Schwartz, D.S.}, year={2008}, month={Sep}, pages={1136–1143} }
 @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{cadenas_maggi_diniz_breitschwerdt_sontakke_breithschwerdt_2007, title={Identification of bacteria from clinical samples using Bartonella alpha-Proteobacteria growth medium}, volume={71}, ISSN={0167-7012}, url={http://dx.doi.org/10.1016/j.mimet.2007.08.006}, DOI={10.1016/j.mimet.2007.08.006}, abstractNote={In an effort to overcome historical problems associated with the isolation of Bartonella species from animal and human blood samples, our laboratory developed a novel, chemically modified, insect-based, liquid culture medium (Bartonella alpha-Proteobacteria growth medium, BAPGM). In this study, we describe the isolation of non-Bartonella bacteria from aseptically obtained human blood and tissue samples that were inoculated into BAPGM pre-enrichment culture medium, and were obtained during attempts to define each individuals Bartonella infection status. After incubation for at least 7 days in liquid BAPGM, pre-enriched inoculums were sub-cultured onto a BAPGM/blood agar plate. Bacterial DNA was extracted from pooled plated colonies and amplified using conventional PCR targeting the 16S rRNA gene. Subsequently, amplicons were cloned, sequenced and compared to GenBank database sequences using the BLAST program. Regardless of the patient's Bartonella status, seventeen samples generated only one 16S rDNA sequence, representing the following genera: Arthrobacter, Bacillus, Bartonella, Dermabacter, Methylobacterium, Propionibacterium, Pseudomonas, Staphylococcus and bacteria listed as “non-cultured” in the GenBank database. Alkalibacterium, Arthrobacter, Erwinia, Kineococcus, Methylobacterium, Propionibacterium, Sphingomonas, and Staphylococcus were isolated from nine Bartonella-infected individuals. Co-isolation of Acinetobacter, Sphingomonas, Staphylococcus spp. and bacteria listed as “non-cultured” in the GenBank database was achieved for four samples in which Bartonella spp. were not detected. Despite the phylogenetic limitations of using partial 16S rRNA gene sequencing for species and strain identification, the investigational methodology described in this study may provide a complementary approach for the isolation and identification of bacteria from patient samples.}, number={2}, journal={Journal of Microbiological Methods}, publisher={Elsevier BV}, author={Cadenas, Maria B. and Maggi, Ricardo G. and Diniz, Pedro P.V.P. and Breitschwerdt, Kyle T. and Sontakke, Sushama and Breithschwerdt, Edward B.}, year={2007}, month={Nov}, pages={147–155} }
 @article{velho_pimentel_negro_okay_diniz_breitschwerdt_2007, title={Severe Anemia, Panserositis, and Cryptogenic Hepatitis in an HIV Patient Infected withBartonella henselae}, volume={31}, ISSN={0191-3123 1521-0758}, url={http://dx.doi.org/10.1080/01913120701696601}, DOI={10.1080/01913120701696601}, abstractNote={Bartonella spp. constitute emerging pathogens of worldwide distribution. Bacillary angiomatosis is the most frequent skin manifestation of bartonelloses; nevertheless, B. henselae infection should always be considered systemic, especially in immunodeficient individuals. The authors report the case of an AIDS patient with bacillary angiomatosis, who had concurrent severe anemia, hepatitis, peritonitis, pleuritis, and pericarditis. Clinical manifestation, electronic microscopic examination of erythrocytes, and histopathology of a papule biopsy suggested a Bartonella sp. infection. Multiple genes were target by PCR and B. henselae DNA was amplified and sequenced (GenBank accession number EF196804) from the angiomatous papule. Treatment with clarithromycin resulted in resolution of the bacillary angiomatosis, fever, anemia, panserosites, and hepatitis.}, number={6}, journal={Ultrastructural Pathology}, publisher={Informa UK Limited}, author={Velho, Paulo Eduardo Neves Ferreira and Pimentel, Vanessa and Negro, Gilda Maria Barbaro Del and Okay, Thelma Suely and Diniz, Pedro Paulo Vissotto de Paiva and Breitschwerdt, Edward Bealmear}, year={2007}, month={Jan}, pages={373–377} }
 @article{de paiva diniz_schwartz_de morais_breitschwerdt_2007, title={Surveillance for Zoonotic Vector-Borne Infections Using Sick Dogs from Southeastern Brazil}, volume={7}, ISSN={1530-3667 1557-7759}, url={http://dx.doi.org/10.1089/vbz.2007.0129}, DOI={10.1089/vbz.2007.0129}, abstractNote={For many vector-borne organisms, dogs can be used as sentinels to estimate the risk of human infection. The objective of this study was to use dogs as sentinels for multiple vector-borne organisms in order to evaluate the potential for human infection with these agents in southeastern Brazil. Blood from 198 sick dogs with clinicopathological abnormalities consistent with tick-borne infections were selected at the São Paulo State University Veterinary Teaching Hospital in Botucatu and tested for DNA and/or antibodies against specific vector-borne pathogens. At least one organism was detected in 88% of the dogs, and Ehrlichia canis DNA was amplified from 78% of the blood samples. Bartonella spp. seroreactivity was found in 3.6%. Leishmania chagasi antibodies were detected in 1% of the dogs. There was no serological or polymerase chain reaction evidence of infection with Anaplasma phagocytophilum, Borrelia burgdorferi, Ehrlichia chaffeensis, Ehrlichia ewingii, and Rickettsia rickettsii. The full E. canis 16S rRNA gene sequence of one of the Brazilian strains obtained in this study was identical to the causative agent of human ehrlichiosis in Venezuela. Ehrlichia canis may pose a human health hazard and may be undiagnosed in southeastern Brazil, whereas exposure to the other organisms examined in this study is presumably infrequent.}, number={4}, journal={Vector-Borne and Zoonotic Diseases}, publisher={Mary Ann Liebert Inc}, author={de Paiva Diniz, Pedro Paulo Vissotto and Schwartz, Denise Saretta and de Morais, Helio Silva Autran and Breitschwerdt, Edward Bealmear}, year={2007}, month={Dec}, pages={689–698} }