@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{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{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{shivaprasad_cadenas_diab_nordhausen_bradway_crespo_breitschwerdt_2008, title={Coxiella-Like Infection in Psittacines and a Toucan}, volume={52}, ISSN={0005-2086 1938-4351}, url={http://dx.doi.org/10.1637/8192-120707-reg}, DOI={10.1637/8192-120707-Reg}, abstractNote={Abstract Seven psittacine birds and a toucan (Ramphastos toco) were diagnosed as infected with Coxiella-like bacteria, based on polymerase chain reaction and bacterial 16S rRNA gene sequence obtained from each bird's liver tissue. Most of the birds exhibited lethargy and weakness for several days prior to death. Gross lesions included mild to moderate emaciation and severely enlarged and mottled pale livers and spleens. Microscopically, there was multifocal necrosis of hepatocytes with infiltration of a mixed population of inflammatory cells, including lymphocytes, heterophils, plasma cells, and macrophages randomly scattered throughout in most birds. In several birds within the macrophages there were vacuoles containing basophilic small cocco-bacilli organisms measuring about 0.5–1 µm. The spleens had increased numbers of mononuclear phagocytic system cells, some of which had vacuoles that contained similar organisms, as observed in the liver. There was inflammation in the epicardium and endocardium, interstitium of the lungs, kidney, adrenal and thyroid glands, lamina propria of the intestine, and in occasional birds in the brain, bursa of Fabricius, and bone marrow associated with similar organisms in the macrophages. Transmission electron microscopy of the liver and lungs in most birds and in the thyroid glands of one bird revealed pleomorphic round to elongated bacteria measuring about 0.45 µm in diameter and more than 1.0 µm in length. Most of these organisms contained a peripheral zone of loosely arranged electron dense material that was located immediately beneath a trilaminar membrane. Occasional organisms contained nucleoids. This is the first documentation of disease presumptively associated with Coxiella-like bacteria in birds. Infección con bacterias parecidas a Coxiella en psitácidos y un tucán. Se diagnosticó la infección con una bacteria parecida a Coxiella en siete aves psitácidas y en un tucán (Ramphastos toco), basándose en una prueba de reacción en cadena por la polimerasa y la secuenciación del gen 16S rRNA obtenido de tejido hepático de cada una de las aves. La mayoría de las aves mostraron letargia y debilidad por varios días antes de la muerte. Las lesiones macroscópicas incluyeron emaciación de leve a moderada, así como hígados y bazos pálidos, moteados y severamente aumentados de tamaño. Microscópicamente, se observó necrosis multifocal de los hepatocitos con infiltración de una mezcla de células inflamatorias incluyendo linfocitos, heterófilos, células plasmáticas y macrófagos distribuidos aleatoriamente en la mayoría de las aves. En varias de las aves, se observaron dentro de los macrófagos hepáticos vacuolas con pequeños organismos basofílicos del tipo cocobacilos que medían entre 0.5 y 1 µm. Los bazos tenían un número aumentado de células del sistema fagocítico, algunas de las cuales mostraban vacuolas que contenían organismos similares a los observados en el hígado. Se observó inflamación asociada a organismos similares en los macrófagos, en el epicardio y endocardio, en el intersticio de los pulmones, riñones, glándulas adrenal y tiroides, en la lámina propia del intestino y en algunas de las aves en el cerebro, bolsa de Fabricio y medula ósea. La microscopía electrónica del hígado y los pulmones en la mayoría de las aves y de la glándula tiroides de un ave, reveló la presencia de bacterias de redondas a alargadas de aproximadamente 0.45 µm de diámetro y más de 1 µm de longitud. La mayoría de estos organismos contenían una zona periférica de material denso localizado inmediatamente por debajo de la membrana trilaminar. Algunos organismos contenían nucléolos. Este es el primer reporte de enfermedad presumiblemente asociada con bacterias parecidas a Coxiella en aves. Abbreviations: bp = base pair; CAHFS = California Animal Health and Food Safety Laboratory System; FA = fluorescent antibody; H&E = hematoxylin and eosin; IHC = immunohistochemistry; NBF = neutral buffered formalin; PAS = periodic acid Schiff; RT-PCR = reverse transcriptase–polymerase chain reaction}, number={3}, journal={Avian Diseases}, publisher={American Association of Avian Pathologists (AAAP)}, author={Shivaprasad, H. L. and Cadenas, M. B. and Diab, S. S. and Nordhausen, R. and Bradway, D. and Crespo, R. and Breitschwerdt, E. B.}, year={2008}, month={Sep}, pages={426–432} }
 @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{valentine_harms_cadenas_birkenheuer_marr_braun-mcneill_maggi_breitschwerdt_2007, title={Bartonella DNA in Loggerhead Sea Turtles}, volume={13}, ISSN={1080-6040 1080-6059}, url={http://dx.doi.org/10.3201/eid1306.061551}, DOI={10.3201/eid1306.061551}, abstractNote={To the Editor: Bartonella are fastidious, aerobic, gram-negative, facultative, intracellular bacteria that infect erythrocytes, erythroblasts, endothelial cells, monocytes, and dendritic cells, and are transmitted by arthropod vectors or by animal scratches or bites (1–6). Currently, 20 species or subspecies of Bartonella have been characterized, of which 8 are known zoonotic pathogens (7). B. henselae has been recently identified from canine blood (8) and from harbor porpoises (9). Pathogenic bacteria are an important threat in terrestrial and marine environments, and in the case of B. henselae, reservoir hosts may be more diverse than currently recognized.}, number={6}, journal={Emerging Infectious Diseases}, publisher={Centers for Disease Control and Prevention (CDC)}, author={Valentine, K. Hope and Harms, Craig A. and Cadenas, Maria B. and Birkenheuer, Adam J. and Marr, Henry S. and Braun-McNeill, Joanne and Maggi, Ricardo G. and Breitschwerdt, Edward B.}, year={2007}, month={Jun}, pages={949–950} }
 @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{rojas_cadenas_lin_benavides_conti_rodriguez-puebla_2007, title={Cyclin D2 and cyclin D3 play opposite roles in mouse skin carcinogenesis}, volume={26}, DOI={10.1038/sj.onc.1209970}, abstractNote={D-type cyclins are components of the cell-cycle engine that link cell signaling pathways and passage throughout G1 phase. We previously described the effects of overexpression cyclin D1, D2 or D3 in mouse epidermis and tumor development. We now asked whether cyclin D2 and/or cyclin D3 play a relevant role in ras-dependent tumorigenesis. Here, we described the effect of cyclin D3 and cyclin D2 overexpression in mouse skin tumor development. Notably, overexpression of cyclin D3 results in reduced tumor development and malignant progression to squamous cell carcinomas (SCC). Biochemical analysis of keratinocytes shows that overexpression of cyclin D3 results in strong reduction of cyclin D2 and its associated kinase activity. Furthermore, we found that reinstatement of cyclin D2 level in the cyclin D3/cyclin D2 bigenic mice results in a complete reversion of the inhibitory action of cyclin D3. Supporting these results, ablation of cyclin D2 results in reduced tumorigenesis and malignant progression. On the other hand, overexpression of cyclin D2 results in an increased number of papillomas and malignant progression. We conclude that cyclin D3 and cyclin D2 play opposite roles in mouse skin tumor development and that the suppressive activity of cyclin D3 is associated with cyclin D2 downregulation.}, number={12}, journal={Oncogene}, author={Rojas, P. and Cadenas, M. B. and Lin, P. C. and Benavides, F. and Conti, C. J. and Rodriguez-Puebla, M. L.}, year={2007}, pages={1723–1730} }
 @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} }