@article{richardson_mainville_arguello-marin_whalley_burton_breitschwerdt_qurollo_2023, title={A second-generation, point-of-care immunoassay provided improved detection of Anaplasma and Ehrlichia antibodies in PCR-positive dogs naturally infected with Anaplasma or Ehrlichia species}, volume={5}, ISSN={["1943-4936"]}, DOI={10.1177/10406387231172723}, abstractNote={ A validated second-generation SNAP 4Dx Plus (Idexx) incorporates new peptides for improved detection of antibodies against Anaplasma and Ehrlichia tick-borne pathogens in dogs. We compared the first- and second-generation SNAP 4Dx Plus using dogs naturally infected with Anaplasma or Ehrlichia species, or dogs seroreactive by an E. canis indirect fluorescent antibody test (IFAT). The second-generation immunoassay was more sensitive than the first-generation for dogs infected with A. phagocytophilum (51.1% and 29.2%, respectively), A. platys (63.6% and 35.3%, respectively), E. canis (96.2% and 88.3%, respectively), or E. ewingii (73.7% and 70.8%, respectively), and for dogs seroreactive by E. canis IFAT (87.3% and 83.9%, respectively). The second-generation immunoassay detected significantly more Anaplasma- or Ehrlichia-infected dogs that were Anaplasma ( p < 0.001) or Ehrlichia ( p = 0.031) seroreactive, respectively, than did the first-generation test. When Ehrlichia seroreactivity by E. canis IFAT and both immunoassays was compared, significantly more E. canis–infected dogs were seroreactive by E. canis IFAT than the first-generation ( p = 0.006) but not the second-generation ( p = 0.125) immunoassay. Significantly more E. ewingii–infected dogs were seroreactive by the first- ( p = 0.011) and second-generation ( p = 0.049) immunoassays than the E. canis IFAT. Medical records available for 7 dogs that were Anaplasma seroreactive by the second-generation but not the first-generation immunoassay revealed case management decisions that might have been different with an immediate anaplasmosis diagnosis, including earlier doxycycline therapy and less hospitalization. The second-generation SNAP 4Dx Plus test offered improved serologic detection of Anaplasma and Ehrlichia in naturally infected dogs. }, journal={JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION}, author={Richardson, Safari S. and Mainville, Celine A. and Arguello-Marin, Andrea and Whalley, Darcy and Burton, Wade and Breitschwerdt, Edward B. and Qurollo, Barbara A.}, year={2023}, month={May} }
 @article{braff_arguello-marin_hanscom_saucier_beall_qurollo_chandrashekar_buch_2023, title={Evaluation of<i> Anaplasma</i> spp. seroprevalence in dogs and association with incidence of human anaplasmosis}, volume={45}, ISSN={["2405-9390"]}, DOI={10.1016/j.vprsr.2023.100923}, abstractNote={Point-of-care (POC) ELISA tests are routinely used in US veterinary practices to screen canine patients for antibodies to tick-transmitted pathogens. Results are also used to monitor spatial and temporal trends in canine seroprevalence, and these data can build awareness of the risk to humans of tick-transmitted diseases such as Lyme disease and anaplasmosis. This study utilized a second-generation test that has incorporated additional Anaplasma-specific peptides into a commercial POC ELISA test to allow detection of Anaplasma spp. antibodies earlier post-infection. A convenience population consisting of 19,894 canine samples from a US commercial diagnostic laboratory were tested using the second-generation POC ELISA test to describe regional Anaplasma spp. canine seroprevalence and assess correlation to anaplasmosis cases reported to Centers for Disease Control and Prevention by state. Antibodies to Anaplasma spp. were detected in 1646 samples (8.3%) with the Northeast and Midwest US census regions having the highest proportion of positive samples. At the state level, a significant correlation was found between canine Anaplasma spp. seroprevalence and human anaplasmosis incidence (r2 = 0.64). Although estimates of canine Anaplasma spp. seroprevalence presented here using the second-generation POC ELISA are generally increased, especially in the Northeast and Midwest, the regional distribution of canine samples testing positive for Anaplasma spp. antibodies is consistent with previous reports. The observed correlation with human anaplasmosis incidence indicates that results from the second-generation POC ELISA will continue to add value in epidemiological assessment of human anaplasmosis risk.}, journal={VETERINARY PARASITOLOGY- REGIONAL STUDIES AND REPORTS}, author={Braff, Jennifer C. and Arguello-Marin, Andrea and Hanscom, Jancy and Saucier, Jill and Beall, Melissa J. and Qurollo, Barbara A. and Chandrashekar, Ramaswamy and Buch, Jesse}, year={2023}, month={Oct} }
 @article{pargass_wint_suepaul_frontera-acevedo_qurollo_2023, title={First reported case of leishmaniasis in a cat in Trinidad and Tobago}, volume={42}, ISSN={["2405-9390"]}, DOI={10.1016/j.vprsr.2023.100896}, abstractNote={A 3-year-old, female, domestic shorthair cat, was presented to the Veterinary Teaching Hospital at the School of Veterinary Medicine (SVM), Trinidad and Tobago for a swollen nose, and multiple, variably sized small masses on both ears. The initial diagnostic tests included a CBC, serum biochemistry profile, cytological evaluation of masses on the ear and nose, and FeLV/FIV testing. The CBC and biochemistry results were unremarkable except for a hyperproteinaemia and hyperglobulinemia. Cytology of the nose and ear lesions revealed mixed inflammation and high numbers of intracellular and extracellular organisms consistent with Leishmania amastigotes. The cat was FeLV/FIV negative. Histopathology and Leishmania IFA and PCR analysis were subsequently performed, confirming the Leishmania diagnosis. The PCR, DNA sequencing and phylogenetic tree analyses identified L. amazonensis. This is the first reported case of L. amazonensis infection in a domestic animal in Trinidad with molecular characterization indicating it exists in the region and is likely being transmitted by sandflies.}, journal={VETERINARY PARASITOLOGY- REGIONAL STUDIES AND REPORTS}, author={Pargass, Indira and Wint, Crystal and Suepaul, Rod and Frontera-Acevedo, Karelma and Qurollo, Barbara A.}, year={2023}, month={Jul} }
 @article{beall_mainville_arguello-marin_clark_lemieux_saucier_thatcher_breitschwerdt_cohn_qurollo_et al._2022, title={An Improved Point-of-Care ELISA for the Diagnosis of Anaplasmosis and Ehrlichiosis During the Acute Phase of Tick-Borne Infections in Dogs}, volume={51}, ISSN={["1946-9837"]}, DOI={10.1016/j.tcam.2022.100735}, abstractNote={Veterinarians often test for serologic evidence of vector-borne infections in sick dogs presenting with clinical signs or to screen for subclinical chronic infections. Additional peptide targets for the detection of antibodies to Anaplasma phagocytophilum, Anaplasma platys, and Ehrlichia canis were added to an existing point-of-care (POC) ELISA test (SNAP 4Dx Plus Test, IDEXX Laboratories, Westbrook, ME). This second-generation, multi-analyte test detects Dirofilaria immitis antigen and antibodies to Anaplasma spp., Borrelia burgdorferi, and Ehrlichia spp. The second-generation test is expected to better meet the needs of practicing veterinarians and their patients. To assess this expectation, the second-generation POC test was evaluated with serum samples from experimentally infected dogs and a broader field population of dogs. Compared to the first-generation test, most dogs experimentally infected with A phagocytophilum (n = 7/8), A platys (n = 4/6), or E canis (n = 4/6) had detectable antibody responses 3-22 days earlier post-infection; these results demonstrated better alignment with polymerase chain reaction (PCR) amplification results and the onset of clinical signs. Using a convenience sample set of 510 sera from both academic and commercial veterinary diagnostic laboratories, the second-generation test had sensitivities greater than 90% for Anaplasma spp. (94.1%), B burgdorferi (95.5%), Ehrlichia spp. (93.4%) and D immitis (98.0%). Specificity ranged from 96.8% - 100% across the four assays. Results from this study demonstrate that the second-generation POC ELISA had an improved ability to detect serologic responses during the acute phase of A phagocytophilum, A platys, and E canis experimental infections. The results from the broader field samples support overall high sensitivity and specificity, consistent with the historical performance of the first-generation POC ELISA test.}, journal={TOPICS IN COMPANION ANIMAL MEDICINE}, author={Beall, Melissa J. and Mainville, Celine A. and Arguello-Marin, Andrea and Clark, Genevieve and Lemieux, Christine and Saucier, Jill and Thatcher, Brendon and Breitschwerdt, Edward B. and Cohn, Leah A. and Qurollo, Barbara A. and et al.}, year={2022} }
 @article{cerreta_yang_ramsay_birkenheuer_rahoi_qurollo_wilson_cushing_2022, title={DETECTION OF VECTOR-BORNE INFECTIONS IN LIONS AND TIGERS AT TWO ZOOS IN TENNESSEE AND OKLAHOMA, USA}, volume={53}, ISSN={["1937-2825"]}, DOI={10.1638/2020-0199}, abstractNote={Abstract: Protozoal and bacterial vector-borne infections are frequently diagnosed in domestic felids. However, with the exception of Mycoplasma haemofelis and Cytauxzoon felis, their occurrence in managed nondomestic felids housed in the United States is largely unknown. Following a case in February 2020 of fulminant cytauxzoonosis in an African lion (Panthera leo), EDTA–whole blood samples were collected opportunistically from February 2020 through June 2020 from 34 adult tigers (Panthera tigris) and eight adult African lions from the same sanctuary in eastern Tennessee as well as 14 adult tigers from a zoo in southern Oklahoma. Samples were analyzed for Cytauxzoon felis, Bartonella spp., hemotropic Mycoplasma, Rickettsia spp., Anaplasma spp., Ehrlichia spp., Babesia spp., and Hepatozoon spp. DNA by PCR amplification. All animals were asymptomatic at the time of collection. None of the Oklahoma animals were positive for vector-borne organisms, but these pathogens were detected in tigers at the Tennessee facility, including Cytauxzoon felis (11.8%), “Candidatus Mycoplasma haemominutum” (5.9%), and Ehrlichia ewingii (2.9%). During the study period, two animals developed clinical signs of cytauxzoonosis and were assessed for vector-borne infections as part of their diagnostic evaluation. This study documents the presence of tick-borne diseases in managed nondomestic felids in the southeastern United States and underscores that ectoparasite control measures should be practiced to minimize exposure of carnivores in managed care.}, number={1}, journal={JOURNAL OF ZOO AND WILDLIFE MEDICINE}, author={Cerreta, Anthony J. and Yang, Tzushan S. and Ramsay, Edward C. and Birkenheuer, Adam J. and Rahoi, Dane and Qurollo, Barbara and Wilson, James and Cushing, Andrew C.}, year={2022}, month={Mar}, pages={50–59} }
 @article{juhasz_wilson_haney_clark_davenport_breitschwerdt_qurollo_2022, title={Rickettsia typhi infection in a clinically-ill dog from Houston, Texas}, volume={35}, ISSN={["2405-9390"]}, DOI={10.1016/j.vprsr.2022.100781}, abstractNote={In 2020, Rickettsia typhi was diagnosed in a dog from Houston, Texas, USA based upon R. typhi IFA seroreactivity in both acute and convalescent sera, and PCR with DNA sequencing of 4 different gene regions, all of which were 100% identical to R. typhi. The dog was clinically ill with intermittent fever, lethargy, inappetence, and lymphadenopathy. Clinicopathological abnormalities included a mild nonregenerative anemia, neutrophilia, lymphopenia, thrombocytopenia, hypoalbuminemia, and elevated ALP. The dog rapidly recovered with doxycycline administration.}, journal={VETERINARY PARASITOLOGY- REGIONAL STUDIES AND REPORTS}, author={Juhasz, Nicholas B. and Wilson, James M. and Haney, Kaitlin N. and Clark, Melissa H. and Davenport, Amy C. and Breitschwerdt, Edward B. and Qurollo, Barbara A.}, year={2022}, month={Oct} }
 @article{kattoor_nikolai_qurollo_wilkes_2022, title={Targeted Next-Generation Sequencing for Comprehensive Testing for Selected Vector-Borne Pathogens in Canines}, volume={11}, ISSN={["2076-0817"]}, DOI={10.3390/pathogens11090964}, abstractNote={The standard for detecting vector-borne pathogens is real-time PCR (rtPCR). However, this requires many individual tests to obtain an accurate diagnosis. The purpose of this study was to develop and validate a targeted next-generation sequencing (NGS) assay for vector-borne pathogens. Pathogen target regions were amplified via PCR using two primer pools that were developed in conjunction with ThermoFisher Scientific, and barcoded DNA libraries were prepared and sequenced with the Ion Torrent S5 system. Data were assembled using SPAdes and mapped to a reference file containing sequences from the pathogens. The raw reads were analyzed to confirm the results. Test feasibility and analytical specificity were evaluated with type strains or validated positive clinical samples from dogs. The analytical sensitivity of the method was compared to Ct values obtained by rtPCR testing. Diagnostic sensitivity and specificity were assessed with a set of known positive and negative clinical samples based on rtPCR testing. Positive and negative percent agreements and Cohen’s kappa were calculated. The primer sets were specific for the intended targets, based on sequence analysis of the amplified products, and the method detected 17 different pathogens. Analytical sensitivity was equivalent to an rtPCR Ct value of approximately 35–36. The positive percent agreement was 92%, and the negative percent agreement was 88%. Cohen’s kappa was 0.804, which indicates almost perfect agreement between the rtPCR assays and the targeted NGS assay. Using a targeted method reduces the costs associated with NGS sequencing and allows for a 2–3 day turn-around time, making this a viable method for detection of vector-borne pathogens in canine whole blood samples.}, number={9}, journal={PATHOGENS}, author={Kattoor, Jobin J. and Nikolai, Emma and Qurollo, Barbara and Wilkes, Rebecca P.}, year={2022}, month={Sep} }
 @article{kidd_hamilton_stine_qurollo_breitschwerdt_2022, title={Vector-borne disease and its relationship to hematologic abnormalities and microalbuminuria in retired racing and show-bred greyhounds}, volume={7}, ISSN={["1939-1676"]}, DOI={10.1111/jvim.16477}, abstractNote={Abstract}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Kidd, Linda and Hamilton, Helen and Stine, Lisa and Qurollo, Barbara and Breitschwerdt, Edward B.}, year={2022}, month={Jul} }
 @article{gin_lashnits_wilson_breitschwerdt_qurollo_2021, title={Demographics and travel history of imported and autochthonous cases of leishmaniosis in dogs in the United States and Canada, 2006 to 2019}, volume={35}, ISSN={["1939-1676"]}, url={https://doi.org/10.1111/jvim.16071}, DOI={10.1111/jvim.16071}, abstractNote={Abstract}, number={2}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, publisher={Wiley}, author={Gin, Taylor Estes and Lashnits, Erin and Wilson, James M. and Breitschwerdt, Edward B. and Qurollo, Barbara}, year={2021}, month={Mar}, pages={954–964} }
 @article{maggi_breitschwerdt_qurollo_miller_2021, title={Development of a Multiplex Droplet Digital PCR Assay for the Detection of Babesia, Bartonella, and Borrelia Species}, volume={10}, ISSN={["2076-0817"]}, url={https://doi.org/10.3390/pathogens10111462}, DOI={10.3390/pathogens10111462}, abstractNote={We describe the development, optimization, and validation of a multiplex droplet digital PCR (ddPCR) assay for the simultaneous detection of Babesia, Bartonella, and Borrelia spp. DNA from several sample matrices, including clinical blood samples from animals and humans, vectors, in-vitro infected human and animal cell lines, and tissues obtained from animal models (infected with Bartonella and/or B. burgdorferi). The multiplex ddPCR assay was able to detect 31 Bartonella, 13 Borrelia, and 24 Babesia species, including Theileria equi, T. cervi, and Cytauxzoon felis. No amplification of Treponema or Leptospira spp. was observed. Sensitivity of 0.2–5 genome equivalent DNA copies per microliter was achieved for different members of the Bartonella and Borrelia genus, depending on the species or matrix type (water or spiked blood DNA) tested. The ddPCR assay facilitated the simultaneous detection of co-infections with two and three vector-borne pathogens comprising four different genera (Babesia, Bartonella, Borrelia, and Theileria) from clinical and other sample sources.}, number={11}, journal={PATHOGENS}, author={Maggi, Ricardo and Breitschwerdt, Edward B. and Qurollo, Barbara and Miller, Jennifer C.}, year={2021}, month={Nov} }
 @article{oney_koo_roy_ren_qurollo_juhasz_vasconcelos_oakley_diniz_2021, title={Evaluation of a commercial microbial enrichment kit used prior DNA extraction to improve the molecular detection of vector-borne pathogens from naturally infected dogs}, volume={188}, ISSN={["1872-8359"]}, DOI={10.1016/j.mimet.2021.106163}, abstractNote={Accurate detection of vector-borne pathogens (VBPs) is extremely important as the number of reported cases in humans and animals continues to rise in the US and abroad. Validated PCR assays are currently the cornerstone of molecular diagnostics and can achieve excellent analytical sensitivity and specificity. However, the detection of pathogens at low parasitemia still presents a challenge for VBP diagnosis, especially given the very low volume of specimens tested by molecular methods. The objective of this study is to determine if a commercially available microbial enrichment kit, used prior DNA extraction, is capable of expanding the overall microbial community and increasing detectable levels of VBPs in canine blood samples through host DNA depletion. This study used EDTA-whole blood samples from dogs naturally infected with varying parasitemia levels of either Anaplasma phagocytophilum, Babesia gibsoni, or Ehrlichia ewingii. For two VBPs, EDTA-blood samples were diluted to determine the effect of microbial concentration at low parasitemia. Paired EDTA-blood samples from each dog were subjected to traditional, automated DNA extraction with or without the microbial concentrating kit (MolYsis®) prior DNA extraction. Relative amounts of pathogen DNA in paired samples were determined by real-time PCR and Next-Generation Sequencing targeting conserved regions of 16S rRNA (for bacteria) and 18S rRNA (for protozoa). Results from the three molecular methods suggest that the microbial concentrating kit did not improve the detection of VBPs, although significantly reduced the presence of host DNA. Alternative methods for VBP enrichment in clinical samples prior to molecular testing should continue to be investigated, as it may significantly improve clinical sensitivity and reduce the number of false-negative results.}, journal={JOURNAL OF MICROBIOLOGICAL METHODS}, author={Oney, Kristina and Koo, Melody and Roy, Chayan and Ren, Songyang and Qurollo, Barbara and Juhasz, Nicholas B. and Vasconcelos, Elton J. R. and Oakley, Brian and Diniz, Pedro P. V. P.}, year={2021}, month={Sep} }
 @article{fricken_qurollo_boldbaatar_wang_jiang_lkhagvatseren_koehler_moore_nymadawa_anderson_et al._2020, title={Genetic diversity of Anaplasma and Ehrlichia bacteria found in Dermacentor and Ixodes ticks in Mongolia}, volume={11}, ISSN={["1877-9603"]}, DOI={10.1016/j.ttbdis.2019.101316}, abstractNote={Anaplasma and Ehrlichia are tick-borne bacterial pathogens that cause human granulocytic anaplasmosis, human monocytic ehrlichiosis, and are severe threats to livestock economies like Mongolia. In this study, ticks were collected, identified, and pooled (n = 299) from three distinct environments across central Mongolia. Each pool was initially tested for Anaplasma/Ehrlichia using a 16S rRNA PCR assay that detects both genera, and specific PCR testing was done to identify those positive samples. Maximum likelihood estimation (MLE) of infection rates of ticks collected from the environment in Selenge aimag (province) found infection rates of Ixodes persulcatus ticks to be 2.0% (95% CI: 0.7, 4.3%) for A. phagocytophilum and 0.8% (95% CI: 0.1, 2.5%) for both nonspecific Ehrlichia and Anaplasma. Ehrlichia muris was only detected in I. persulcatus ticks collected from the Selenge aimag, where the MLE was 1.2% (95% CI: 0.1, 2.5%). The calculated MLE infection rate of Anaplasma spp. in questing Dermacentor nuttalli ticks ranged from 1.9% (95% CI: 1.1, 9.1%) in the Tov aimag to 2.3% (95% CI: 1.3, 10.8%) in the Selenge aimag. However, when examining MLE in ticks removed from livestock, estimates increase substantially, ranging from 7.8% (95% CI: 4.2, 13.3%) in Dornogovi to 22.5% (95% CI: 14.3, 34.3%) in Selenge, suggesting that livestock play a key role in disease maintenance. Considering the collective economic losses that can result from these pathogens and the potential for illness in nomadic herdsmen, these results highlight the need for enhanced TBD surveillance and prevention measures within Mongolia.}, number={1}, journal={TICKS AND TICK-BORNE DISEASES}, author={Fricken, Michael E. and Qurollo, Barbara A. and Boldbaatar, Bazartseren and Wang, Ya-Wei and Jiang, Rui-Ruo and Lkhagvatseren, Sukhbaatar and Koehler, Jeffrey W. and Moore, Thomas C. and Nymadawa, Pagbajab and Anderson, Benjamin D. and et al.}, year={2020}, month={Jan} }
 @article{tyrrell_qurollo_mowat_kennedy-stoskopf_2020, title={MOLECULAR PREVALENCE OF SELECTED VECTOR-BORNE ORGANISMS IN CAPTIVE RED WOLVES (CANIS RUFUS)}, volume={51}, ISSN={["1937-2825"]}, DOI={10.1638/2019-0162}, abstractNote={Abstract: The red wolf (Canis rufus) is a critically endangered North American canid, with surviving conspecifics divided between a captive breeding population and a reintroduced free-ranging population. The goal of this study was to assess the prevalence of selected vector-borne pathogens in captive red wolves. Whole blood samples were collected from 35 captive red wolves. Quantitative polymerase chain reaction (PCR) assays were performed on extracted DNA to identify infection by Trypanosoma cruzi and vector-borne organisms within the following genera: Anaplasma, Babesia, Bartonella, Ehrlichia, Mycoplasma, Neoehrlichia, Neorickettsia, and Rickettsia. All red wolves sampled were PCR-negative for all tested organisms. These pathogens are unlikely to constitute threats to red wolf conservation and breeding efforts under current captive management conditions. The results of this study establish a baseline that may facilitate ongoing disease monitoring in this species.}, number={3}, journal={JOURNAL OF ZOO AND WILDLIFE MEDICINE}, author={Tyrrell, Jeffrey D. and Qurollo, Barbara A. and Mowat, Freya M. and Kennedy-Stoskopf, Suzanne}, year={2020}, month={Sep}, pages={663–667} }
 @article{fricken_voorhees_koehler_asbun_lam_qurollo_hogan_baasandagva_jigjav_schoepp_2020, title={Molecular Characteristics of Rickettsia in Ticks Collected along the Southern Border of Mongolia}, volume={9}, ISSN={["2076-0817"]}, DOI={10.3390/pathogens9110943}, abstractNote={Tick-borne infections are a significant threat to public health, particularly in regions where individuals frequently enter tick habitats. Roughly 26% of the population in Mongolia practice nomadic pastoralism and are considered at high risk of exposure to ticks and the diseases they carry. This study tested ticks from Mongolia’s southern border for Rickettsia spp. to better understand the epidemiology of tick-borne diseases in the region. Dermacentor nuttalli and Hyalomma asiaticum ticks (n = 4022) were pooled and tested for Rickettsia spp. by real-time PCR. Melt-curve analyses and Sanger sequencing were used to identify Rickettsia species. Approximately 64% of the 786 tick pools tested positive for Rickettsia bacteria. Melt curve analyses identified four different Rickettsia species circulating in these tick pools. Amplicon sequencing of the ompA gene identified Rickettsia spp. that closely resembled R. raoultii and R. sibirica. Dermacentor nuttalli ticks from Govi-Altai had the highest maximum likelihood estimation infection rate 48.4% (95% CI: 41.7–56.5%), while Hyalommaasiaticum collected in Omnogovi had a rate of 7.6% (95% CI: 6.2–9.2%). The high detection of Rickettsia suggests a substantial risk of infection in southern Mongolia. Further studies are necessary to investigate the clinical burden of tick-borne diseases in Mongolia.}, number={11}, journal={PATHOGENS}, author={Fricken, Michael E. and Voorhees, Matthew A. and Koehler, Jeffrey W. and Asbun, Carmen and Lam, Brandon and Qurollo, Barbara and Hogan, Kathryn M. and Baasandagva, Uyanga and Jigjav, Battsetseg and Schoepp, Randal J.}, year={2020}, month={Nov} }
 @article{wilson_breitschwerdt_juhasz_marr_galvao_pratt_qurollo_2020, title={Novel Rickettsia Species Infecting Dogs, United States}, volume={26}, ISSN={["1080-6059"]}, DOI={10.3201/eid2612.200272}, abstractNote={In 2018 and 2019, spotted fever was suspected in 3 dogs in 3 US states. The dogs had fever and hematological abnormalities; blood samples were Rickettsia seroreactive. Identical Rickettsia DNA sequences were amplified from the samples. Multilocus phylogenetic analysis showed the dogs were infected with a novel Rickettsia species related to human Rickettsia pathogens.}, number={12}, journal={EMERGING INFECTIOUS DISEASES}, author={Wilson, James M. and Breitschwerdt, Edward B. and Juhasz, Nicholas B. and Marr, Henry S. and Galvao, Joao Felipe de Brito and Pratt, Carmela L. and Qurollo, Barbara A.}, year={2020}, month={Dec}, pages={3011–3015} }
 @article{qurollo_buch_chandrashekar_beall_breitschwerdt_yancey_caudill_comyn_2019, title={Clinicopathological findings in 41 dogs (2008‐2018) naturally infected with
            Ehrlichia ewingii}, volume={33}, ISSN={0891-6640 1939-1676}, url={http://dx.doi.org/10.1111/jvim.15354}, DOI={10.1111/jvim.15354}, abstractNote={BackgroundEhrlichia ewingii is the most seroprevalent Ehrlichia‐infecting dogs in the southern and mid‐western United States. Fever, lameness, and polyarthritis are commonly reported findings in dogs naturally infected with E. ewingii.}, number={2}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Qurollo, Barbara A. and Buch, Jesse and Chandrashekar, Ramaswamy and Beall, Melissa J. and Breitschwerdt, Edward B. and Yancey, Caroline B. and Caudill, Alexander H. and Comyn, Alaire}, year={2019}, month={Jan}, pages={618–629} }
 @article{qurollo_2019, title={Feline Vector-Borne Diseases in North America}, volume={49}, ISSN={["1878-1306"]}, DOI={10.1016/j.cvsm.2019.02.012}, abstractNote={“In North America, with the exceptions of Bartonella henselae and Cytauxzoon felis, feline vector-borne diseases (FVBDs) have been minimally studied in domestic cats. Cats can be infected with many of the same vector-borne pathogens that infect dogs. Nonspecific clinical signs linked to FVBDs and low prevalence of certain vector-borne pathogens contribute to a limited awareness of FVBDs in sick cats. As clinicians become informed about FVBDs and as vector-borne disease diagnostics are routinely applied to evaluate sick cats, we will gain a stronger understanding of vector-borne pathogens in cats. This article focuses on recent findings related to FVBDs.”}, number={4}, journal={VETERINARY CLINICS OF NORTH AMERICA-SMALL ANIMAL PRACTICE}, author={Qurollo, Barbara}, year={2019}, month={Jul}, pages={687-+} }
 @article{tyrrell_qurollo_tornquist_schlaich_kelsey_chandrashekar_breitschwerdt_2019, title={Molecular identification of vector-borne organisms in Ehrlichia seropositive Nicaraguan horses and first report of Rickettsia felis infection in the horse}, volume={200}, ISSN={["1873-6254"]}, DOI={10.1016/j.actatropica.2019.105170}, abstractNote={Certain vector-borne organisms serve as etiological agents of equine disease. After previously identifying a new Ehrlichia species in horses from Mérida, we aimed to determine the infection frequency and screen for a wide range of vector-borne organisms from 93 tick-exposed, Ehrlichia seropositive horses in this region. PCR assays were performed to identify infection by organisms within the following genera: Anaplasma, Babesia, Bartonella, Ehrlichia, Leishmania, Mycoplasma, Neorickettsia, Rickettsia and Theileria. Overall, 90/93 horses (96.8%) were infected with one or more vector-borne organisms. Ninety (96.8%) horses were infected with Theileria equi and 21 (26.8%) with Babesia caballi. Nine (9.7%) horses were infected with the novel Ehrlichia species previously designated H7, reported in horses from Nicaragua and Brazil. Two horses (2.2%) were infected with Rickettsia felis. Anaplasma, Bartonella, Leishmania, Mycoplasma, or Neorickettsia species DNA was not amplified from any horse. Ticks collected from horses infected with vector-borne organisms were identified as Amblyomma cajennense sensu lato and Dermacentor nitens. Horses in Mérida are infected by a range of vector-borne organisms, including B. caballi, T. equi, Ehrlichia species H7, and R. felis. To the authors' knowledge, this constitutes the first report of molecular detection of R. felis in horses.}, journal={ACTA TROPICA}, author={Tyrrell, Jeffrey D. and Qurollo, Barbara A. and Tornquist, Susan J. and Schlaich, Kathryn G. and Kelsey, Jennifer and Chandrashekar, Ramaswamy and Breitschwerdt, Edward B.}, year={2019}, month={Dec} }
 @article{birkenheuer_marr_wilson_breitschwerdt_qurollo_2018, title={Babesia gibsoni
 cytochrome b mutations in canine blood samples submitted to a US veterinary diagnostic laboratory}, volume={32}, ISSN={0891-6640}, url={http://dx.doi.org/10.1111/jvim.15300}, DOI={10.1111/jvim.15300}, abstractNote={
Background: Babesiosis caused by Babesia gibsoni is recognized throughout the world and can be difficult to treat. Resistance to atovaquone is associated with mutations in the B. gibsoni mitochondrial genome, specifically the M128 position of cytochrome b (cytb). The prevalence of cytb mutations in North America has not been reported.}, number={6}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Birkenheuer, Adam J. and Marr, Henry S. and Wilson, James M. and Breitschwerdt, Edward B. and Qurollo, Barbara A.}, year={2018}, month={Oct}, pages={1965–1969} }
 @article{qurollo_larsen_rakotondrainibe_mahefarisoa_rajaonarivelo_razafindramanana_breitschwerdt_junge_williams_2018, title={Molecular surveillance of novel tick-borne organisms in Madagascar's lemurs}, volume={9}, ISSN={["1877-9603"]}, DOI={10.1016/j.ttbdis.2018.02.012}, abstractNote={The discovery and characterization of emerging tick-borne organisms are critical for global health initiatives to improve animal and human welfare (One Health). It is possible that unknown tick-borne organisms underlie a subset of undiagnosed illness in wildlife, domesticated species, and humans. Our study lends support to the One Health concept by highlighting the prevalence of three blood-borne organisms in wild lemurs living in close proximity to domesticated species and humans. Previously, our team identified three novel, presumably tick-borne, intravascular organisms, belonging to the genera Babesia, Borrelia, and Neoehrlichia, circulating in two of Madagascar's lemur species. Here, we extend our previous observation by developing a targeted molecular surveillance approach aimed at determining the prevalence of these organisms in lemurs. Using quantitative PCR, we provide Babesia, Borrelia, and Neoehrlichia prevalence data for 76 individuals comprising four lemur species located in eastern Madagascar. Our results indicate a high prevalence (96%) of Babesia across sampled individuals with lower prevalences for Neoehrlichia (36%) and Borrelia (14.5%). In light of our results, we recommend additional studies of these tick-borne organisms to determine pathogenicity and assess zoonotic potency to other animals and humans in Madagascar.}, number={3}, journal={TICKS AND TICK-BORNE DISEASES}, author={Qurollo, Barbara A. and Larsen, Peter A. and Rakotondrainibe, Hajanirina H. and Mahefarisoa, Karine and Rajaonarivelo, Tsiky and Razafindramanana, Josia and Breitschwerdt, Edward B. and Junge, Randall E. and Williams, Cathy V.}, year={2018}, month={Mar}, pages={672–677} }
 @article{vieira_qurollo_mongruel_baggio_vidotto_breitschwerdt_vieira_2018, title={Potentially Same Novel Ehrlichia Species in Horses in Nicaragua and Brazil}, volume={24}, ISSN={1080-6040 1080-6059}, url={http://dx.doi.org/10.3201/eid2405.172076}, DOI={10.3201/eid2405.172076}, abstractNote={To the}, number={5}, journal={Emerging Infectious Diseases}, publisher={Centers for Disease Control and Prevention (CDC)}, author={Vieira, Thállitha S.W.J. and Qurollo, Barbara A. and Mongruel, Anna C.B. and Baggio, Rafael A. and Vidotto, Odilon and Breitschwerdt, Edward B. and Vieira, Rafael F.C.}, year={2018}, month={May}, pages={953–953} }
 @article{lado_qurollo_williams_junge_klompen_2018, title={The microbiome of Haemaphysalis lemuris (Acari: Ixodidae), a possible vector of pathogens of endangered lemur species in Madagascar}, volume={9}, ISSN={["1877-9603"]}, DOI={10.1016/j.ttbdis.2018.05.003}, abstractNote={Lemurs are primate species that are endemic to Madagascar. At present, about 90% of lemur species are endangered, and 5 species are among the 25 most endangered primates worldwide. Health status is a major factor impacting the viability of wild populations of many endangered species including lemurs. Given this context, we analyzed the microbiome of 24 specimens of Haemaphysalis lemuris, the most common tick parasitizing lemurs in their native habitats. Ticks were collected from 6 lemur species and microbiomes analyzed using next-generation sequencing. Our results show that the H. lemuris microbiome is highly diverse, including over 500 taxa, 267 of which were identified to genus level. Analysis of the microbiome also shows that there is a distinct “host” (lemur species) component when explaining the differences among and between microbial communities of H. lemuris. This “host” component seems to overwhelm any “locality” (geographic origin of the sample) component. In addition to the microbiome data, targeted PCR was used to test for the presence of three pathogens recently detected in the blood of wild lemurs: Borrelia sp., Candidatus Neoehrlichia sp., and Babesia sp. Overall, the presence of DNA of Rickettsia spp., Bartonella spp., Francisella spp., and a Babesia sp., in H. lemuris, is consistent with the hypothesis that these ectoparasites may act as vector for these pathogens. Further studies assessing vector competence are needed to confirm this hypothesis.}, number={5}, journal={TICKS AND TICK-BORNE DISEASES}, author={Lado, Paula and Qurollo, Barbara and Williams, Cathy and Junge, Randall and Klompen, Hans}, year={2018}, month={Jul}, pages={1252–1260} }
 @article{yancey_diniz_breitschwerdt_hegarty_wiesen_qurollo_2017, title={Doxycycline treatment efficacy in dogs with naturally occurring Anaplasma phagocytophilum
 infection}, volume={59}, ISSN={0022-4510}, url={http://dx.doi.org/10.1111/jsap.12799}, DOI={10.1111/jsap.12799}, abstractNote={ObjectivesTo evaluate doxycycline treatment efficacy and post‐treatment pathogen persistence in dogs naturally infected with Anaplasma phagocytophilum in endemic regions of the USA.}, number={5}, journal={Journal of Small Animal Practice}, publisher={Wiley}, author={Yancey, C. B. and Diniz, P. P. V. P. and Breitschwerdt, E. B. and Hegarty, B. C. and Wiesen, C. and Qurollo, B. A.}, year={2017}, month={Dec}, pages={286–293} }
 @article{qurollo_archer_schreeg_marr_birkenheuer_haney_thomas_breitschwerdt_2017, title={Improved molecular detection of Babesia infections in animals using a novel quantitative real-time PCR diagnostic assay targeting mitochondrial DNA}, volume={10}, ISSN={1756-3305}, url={http://dx.doi.org/10.1186/s13071-017-2064-1}, DOI={10.1186/s13071-017-2064-1}, abstractNote={Babesiosis is a protozoal, tick transmitted disease found worldwide in humans, wildlife and domesticated animals. Commonly used approaches to diagnose babesiosis include microscopic examination of peripheral blood smears, detection of circulating antibodies and PCR. To screen and differentiate canine Babesia infections many PCR assays amplify the 18S rRNA gene. These sequences contain hypervariable regions flanked by highly conserved regions allowing for amplification of a broad-range of Babesia spp. However, differences in the 18S rRNA gene sequence of distantly related clades can make it difficult to design assays that will amplify all Babesia species while excluding the amplification of other eukaryotes. By targeting Babesia mitochondrial genome (mtDNA), we designed a novel three primer qPCR with greater sensitivity and broader screening capabilities to diagnose and differentiate Babesia spp. Using 13 Babesia mtDNA sequences, a region spanning two large subunit rRNA gene fragments (lsu5-lsu4) was aligned to design three primers for use in a qPCR assay (LSU qPCR) capable of amplifying a wide range of Babesia spp. Plasmid clones were generated and used as standards to determine efficiency, linear dynamic range and analytical sensitivity. Animals naturally infected with vector-borne pathogens were tested retrospectively and prospectively to determine relative clinical sensitivity and specificity by comparing the LSU qPCR to an established 18S rDNA qPCR. The LSU qPCR efficiencies ranged between 92 and 100% with the limit of detection at five copies/reaction. The assay did not amplify mammalian host or other vector-borne pathogen gDNA except Cytauxzoon felis (a feline protozoal pathogen). The LSU qPCR assay amplified 12 different Babesia. sp. and C. felis from 31/31 (100%) archived samples, whereas the 18S qPCR amplified only 26/31 (83.9%). By prospective analysis, 19/394 diagnostic accessions (4.8%) were LSU qPCR positive, compared to 11/394 (2.8%) 18S rDNA qPCR positive. We have developed a more sensitive qPCR assay with a more expansive range of Babesia spp. detection by targeting a highly conserved region of mtDNA, when compared to an established 18S qPCR.}, number={1}, journal={Parasites & Vectors}, publisher={Springer Nature}, author={Qurollo, Barbara A. and Archer, Nikole R. and Schreeg, Megan E. and Marr, Henry S. and Birkenheuer, Adam J. and Haney, Kaitlin N. and Thomas, Brittany S. and Breitschwerdt, Edward B.}, year={2017}, month={Mar} }
 @article{kidd_qurollo_lappin_richter_hart_hill_osmond_breitschwerdt_2017, title={Prevalence of Vector-Borne Pathogens in Southern California Dogs With Clinical and Laboratory Abnormalities Consistent With Immune-Mediated Disease}, volume={31}, ISSN={0891-6640}, url={http://dx.doi.org/10.1111/jvim.14735}, DOI={10.1111/jvim.14735}, abstractNote={BackgroundStudies investigating the prevalence of vector‐borne pathogens in southern California dogs are limited. Occult infections might be misdiagnosed as idiopathic immune‐mediated disease.}, number={4}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Kidd, L. and Qurollo, B. and Lappin, M. and Richter, K. and Hart, J.R. and Hill, S. and Osmond, C. and Breitschwerdt, E.B.}, year={2017}, month={May}, pages={1081–1090} }
 @article{foley_stephenson_cubilla_qurollo_breitschwerdt_2016, title={A putative marker for human pathogenic strains of Anaplasma phagocytophilum correlates with geography and host, but not human tropism}, volume={7}, ISSN={1877-959X}, url={http://dx.doi.org/10.1016/j.ttbdis.2015.12.015}, DOI={10.1016/j.ttbdis.2015.12.015}, abstractNote={Anaplasma phagocytophilum is an Ixodes species tick-transmitted bacterium that is capable of infecting a variety of host species, although there is a diversity of bacterial strains with differing host tropism. Recent analysis of A. phagocytophilum strains suggested that "drhm", a gene locus designated "distantly related to human marker" (drhm), which was predicted to be an integral membrane protein with possible transporter functions was not present in available canine and human isolates. By assessing 117 strains from 14 host species from across the US, we extended this analysis. Phylogenetic clades were associated with geography, but not host species. Additionally, a virulent clade that lacks drhm and infects dogs, horses, and humans in northeastern US was identified.}, number={2}, journal={Ticks and Tick-borne Diseases}, publisher={Elsevier BV}, author={Foley, Janet and Stephenson, Nicole and Cubilla, Michelle Pires and Qurollo, Barbara and Breitschwerdt, Edward B.}, year={2016}, month={Mar}, pages={390–393} }
 @article{meichner_qurollo_anderson_grindem_savage_breitschwerdt_2015, title={Naturally OccurringEhrlichia ewingiiandMycoplasmasp. Co-Infection in a Goat}, volume={29}, ISSN={0891-6640}, url={http://dx.doi.org/10.1111/jvim.13644}, DOI={10.1111/jvim.13644}, abstractNote={A 9-year-old nonpregnant, nonlactating doe Boer goat was examined because of a 2-day history of not being able to stand. Other than diarrhea associated with coccidiosis in the first year of life, the goat did not have a history of illness. The owner had obtained the goat at approximately 2 months of age. The goat lived with 3 other goats in the same pen on the same premise located on the coastal plains of North Carolina. The goats spent the majority of time in a barn, with access to a wooded 1-acre lot where they browsed. Routine deworming prophylaxis was verified by fecal egg counts. The goat was vaccinated for clostridial diseases, was fed 1 cup of 13.5% protein commercial goat pellets twice a day, and had free access to good quality coastal Bermuda hay. At presentation for recumbency, the goat was nonweight bearing on the right forelimb and could stand only with assistance, but was unable to walk. Otherwise, the goat was bright, alert, responsive, and had a good appetite. Body condition score (5/5), body weight (65.5 kg), rectal temperature (39.2°C [102.5°F]), heart rate (80 beats per minute), respiratory rate (24 breaths per minute), mucous membrane color, and capillary refill time were normal. An abscess was present on the ventral aspect of the mammary gland. To further assess the lameness, lateral, craniocaudal, and oblique radiographs of the right humerus were obtained, and a mildly comminuted, moderately proximo-caudally and medially displaced short oblique fracture of the proximal humeral diaphysis was identified (Fig 1). Marked soft tissue swelling was associated with the fracture. Mild rounding and blunting of the fracture margins without evidence of callus formation were observed and, consequently, some degree of chronicity (>7–10 days) was considered likely. A CBC identified mild macrocytic normochromic anemia (PCV, 20%; reference range, 22–38%; mean cell volume [MCV], 26.4 fL; reference range, 16–24 fL; mean corpuscular hemoglobin concentration [MCHC],}, number={6}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Meichner, K. and Qurollo, B.A. and Anderson, K.L. and Grindem, C.B. and Savage, M. and Breitschwerdt, E.B.}, year={2015}, month={Oct}, pages={1735–1738} }
 @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{qurollo_chandrashekar_hegarty_beall_stillman_liu_thatcher_pultorak_cerrito_walsh_et al._2014, title={A serological survey of tick-borne pathogens in dogs in North America and the Caribbean as assessed by Anaplasma phagocytophilum, A. platys, Ehrlichia canis, E. chaffeensis, E. ewingii, and Borrelia burgdorferi species-specific peptides.}, volume={4}, DOI={10.3402/iee.v4.24699.}, abstractNote={{"Label"=>"INTRODUCTION", "NlmCategory"=>"BACKGROUND"} Tick-borne pathogens cause a spectrum of disease manifestations in both dogs and humans. Recognizing regional and temporal shifts in exposure are important as tick distributions change. To better delineate regional exposure to canine tick-borne pathogens, an expanded set of species-specific peptides were used to detect Anaplasma phagocytophilum (Aph), Anaplasma platys (Apl), Ehrlichia canis (Ec), Ehrlichia chaffeensis (Ech), Ehrlichia ewingii (Eew), and Borrelia burgdorferi (Bb) antibodies in canine serum. {"Label"=>"METHODS", "NlmCategory"=>"METHODS"} Archived canine serum samples (n=6,582) collected during 2008-2010 and in 2012 from the US, Canada, and the Caribbean were retrospectively screened for antibodies against Ehrlichia and Anaplasma species-specific peptides. Overall, regional and temporal seroprevalence rates were determined. {"Label"=>"RESULTS", "NlmCategory"=>"RESULTS"} Overall Bb and Eew were the most seroprevalent pathogens. During 2008-2010, seroprevalence rates increased overall for Aph and Ech, and regionally, Bb and Aph seroprevalence rates increased in the South. Canada had unexpectedly high seroprevalence rates for Ec and Apl. The most common co-exposures were Eew+Ech, followed by Aph+Bb and Eew+Bb. {"Label"=>"CONCLUSIONS", "NlmCategory"=>"CONCLUSIONS"} This study demonstrated significant shifts in canine vector-borne disease seroprevalence rates. The use of specific peptides facilitated improved geographic delineation of tick-borne pathogen distributions among dogs, which may enhance epidemiological surveillance of vector-borne pathogens shared by dogs and humans.}, number={1}, journal={Infection Ecology & Epidemiology}, author={Qurollo, B.A. and Chandrashekar, R. and Hegarty, B.C. and Beall, M.J. and Stillman, B.A. and Liu, J. and Thatcher, B. and Pultorak, E. and Cerrito, B. and Walsh, M. and et al.}, year={2014}, month={Jan}, pages={24699} }
 @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{qurollo_balakrishnan_cannon_maggi_breitschwerdt_2014, title={Co-infection with Anaplasma platys, Bartonella henselae, Bartonella koehlerae and ‘Candidatus Mycoplasma haemominutum’ in a cat diagnosed with splenic plasmacytosis and multiple myeloma}, volume={16}, ISSN={1098-612X 1532-2750}, url={http://dx.doi.org/10.1177/1098612X13519632}, DOI={10.1177/1098612x13519632}, abstractNote={Anaplasma platys ( Apl), ‘ Candidatus Mycoplasma haemominutum’ ( CMh), Bartonella henselae ( Bh) and Bartonella koehlerae ( Bk) were confirmed by polymerase chain reaction (PCR) amplification and DNA sequencing in a cat diagnosed with multiple myeloma. Other inconsistently documented hematologic abnormalities included anemia, thrombocytopenia, eosinophilia and hypoglycemia. Persistent Apl infection was confirmed for the first time in a North American cat by sequencing three bacterial genes ( 16S rRNA, p44 and GroEL) in peripheral blood samples collected 100 days apart. Following doxycycline treatment for Apl, multiple myeloma was diagnosed based upon a monoclonal gammopathy and splenic plasmacytosis, and the cat was treated with melphalan, chlorambucil and prednisolone. Apl DNA was not amplified from post-treatment blood samples and the hyperglobulinemia resolved temporarily following chemotherapy. Retrospective PCR analysis of stored DNA extracts identified CMh, Bk and Bh infections. Retrospective PCR for antigen receptor rearrangements (PARR) of splenic aspirates did not confirm B- or T-cell clonality. Co-infection with multiple vector-borne pathogens should be a diagnostic consideration in cats with chronic hypergammaglobulinemia, monoclonal gammopathy and splenic plasmacytosis.}, number={8}, journal={Journal of Feline Medicine and Surgery}, publisher={SAGE Publications}, author={Qurollo, Barbara A and Balakrishnan, Nandhakumar and Cannon, Coralie Zegre and Maggi, Ricardo G and Breitschwerdt, Edward B}, year={2014}, month={Jan}, pages={713–720} }
 @article{qurollo_riggins_comyn_zewde_breitschwerdt_2014, title={Development and Validation of a Sensitive and Specific sodB-Based Quantitative PCR Assay for Molecular Detection of Ehrlichia Species}, volume={52}, ISSN={0095-1137 1098-660X}, url={http://dx.doi.org/10.1128/jcm.02340-14}, DOI={10.1128/jcm.02340-14}, abstractNote={ABSTRACT}, number={11}, journal={Journal of Clinical Microbiology}, publisher={American Society for Microbiology}, author={Qurollo, B. A. and Riggins, D. and Comyn, A. and Zewde, M. T. and Breitschwerdt, E. B.}, year={2014}, month={Aug}, pages={4030–4032} }
 @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={BackgroundFrequent 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.}, 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{dantas-torres_capelli_giannelli_ramos_lia_cantacessi_de caprariis_de tommasi_latrofa_lacasella_et al._2013, title={Efficacy of an imidacloprid/flumethrin collar against fleas, ticks and tick-borne pathogens in dogs}, volume={6}, ISSN={1756-3305}, url={http://dx.doi.org/10.1186/1756-3305-6-245}, DOI={10.1186/1756-3305-6-245}, abstractNote={Abstract}, number={1}, journal={Parasites & Vectors}, publisher={Springer Nature}, author={Dantas-Torres, Filipe and Capelli, Gioia and Giannelli, Alessio and Ramos, Rafael Antonio and Lia, Riccardo and Cantacessi, Cinzia and de Caprariis, Donato and De Tommasi, Anna and Latrofa, Maria and Lacasella, Vita and et al.}, year={2013}, pages={245} }
 @article{qurollo_davenport_sherbert_grindem_birkenheuer_breitschwerdt_2013, title={Infection with Panola MountainEhrlichiasp. in a Dog with Atypical Lymphocytes and Clonal T-Cell Expansion}, volume={27}, ISSN={0891-6640}, url={http://dx.doi.org/10.1111/jvim.12148}, DOI={10.1111/jvim.12148}, abstractNote={An 11-year-old, castrated male Scottish Terrier from Raleigh, NC that lived predominantly indoors and had no travel history was referred to the North Carolina State University Veterinary Health Complex (NCSU-VHC) in October 2012 for routine reevaluation of hepatobiliary disease. The dog's previous 3-year medical history included biliary mucocele (2010); neutrophilic hepatitis (2011); recurrent Escherichia coli urinary tract infections, esophageal dysmotility, aspiration pneumonia, and transient thrombocytopenia (2012); and food responsive enteropathy (2009–2012). All of these medical problems were well controlled at the time of examination and no clinical abnormalities were reported by the dog's owners. In the months before the present examination, ticks occasionally were noted and fleas were commonly found on the dog despite reported use of preventive therapies. On physical examination, the dog was obese (body condition score, 7 out of 9) and had mild hepatomegaly, both of which had been present for more than a year. Notable CBC findings included mild thrombocytopenia (platelet count, 143,000/μL; reference interval [RI], 190,000–468,000/μL) and an increased number of atypical lymphocytes (1,773/μL), with normal appearing lymphocytes within the laboratory reference range (1,854/μL; RI, 594–3,305/μL) and an otherwise normal differential cell count. A review of the blood smear by a pathologist identified a population of immature lymphocytes with angular nuclei and cell shape, multiple nucleoli, and deeply basophilic, vacuolated cytoplasm that had a tendency to mold into surrounding cells (Fig 1). These morphologic abnormalities raised the suspicion of possible lymphoid neoplasia, although these changes can be seen secondary to reactive processes such as chronic inflammatory or infectious diseases. Serum biochemical abnormalities included an increase in alkaline phosphatase activity (ALP; 817 IU/L; RI, 16–140 IU/L) and alanine aminotransferase activity (ALT; 60 IU/L; RI, 12–54). During the previous 12-month period, ALT activity varied between normal and 80 IU/L and ALP activity varied between 359 and 534 IU/L. Because these hematological and serum biochemical abnormalities were present 2 weeks later, abdominal ultrasound examination was repeated and identified mottled splenic parenchyma and several previously identified changes including hyperechoic nodular hepatomegaly; mild dystrophic mineralization of the spleen, liver, kidneys, and prostate; and, a thickened urinary bladder apex. Splenic cytology identified a mixed lymphoid population and mild extramedullary hematopoiesis. Liver cytology identified normal hepatocytes and an expansion of intermediate lymphocytes, raising concern for lymphoma. Cytology of a palpably normal popliteal lymph node identified expansion of intermediate to large lymphocytes, most consistent with lymphoma (Fig 2). T-cell clonality was documented in a liver aspirate after submission to NCSU-CVM Clinical Immunology Laboratory for polymerase chain reaction (PCR) to identify antigen receptor rearrangements (PARR). Aseptically collected ethylenediamine tetraacetic acid (EDTA)-anticoagulated whole blood and serum were submitted to the NCSU-CVM Vector Borne Diseases Diagnostic Laboratory (VBDDL) for a vector-borne pathogen serology panel and Bartonella alpha-Proteobacteria Growth Medium (BAPGM) enrichment blood culture PCR platform. The dog was seronegative for antibodies to Anaplasma phagocytophilum, Anaplasma platys, and Borrelia burgdorferi and for Dirofilaria immitis antigen using a commercial enzyme-linked immunosorbent assay based kit (SNAP® 4Dx® Plusa); the dog was seronegative for antibodies to Babesia canis, Babesia gibsoni, Bartonella henselae, and Bartonella vinsonii subsp. berkhoffii, using indirect immunofluorescent antibody (IFA) testing. By IFA, the dog was seroreactive to Rickettsia rickettsii (1:64; laboratory cut-off value, 1:64) and Ehrlichia canis (1:1028; laboratory cut-off value, 1:64) antigens, but seronegative to Ehrlichia spp. peptides in the SNAP 4Dx Plusa kit, which is known to detect antibodies against E. canis, E. chaffeensis, and E. ewingii. Bartonella enrichment blood culture and PCR were negative. Nine months before this presentation, the dog was seronegative in the NCSU-CVM-VBDDL to the vector-borne pathogens tested above. Because of the discrepancy between the E. canis IFA and SNAP 4Dx Plusa test results, Ehrlichia spp. PCR was performed on DNA extracted from the dog's EDTA-anticoagulated whole blood using a previously described 16S ribosomal RNA PCR assay.1 A 351 base pair portion of the 16S rRNA gene was amplified, sequenced, and investigated in the NCBI GenBank nucleotide database. The partial 16S rRNA gene identified infection with the Panola Mountain Ehrlichia sp. (PME) with 100% coverage (331 bp) and 100% identity to PME 16S ribosomal RNA gene (DQ324367.1). To confirm these results, alternative PME genes, including gltA and map1 were targeted. Furthermore, sodb, a gene not previously sequenced for PME, was amplified using newly developed primers designed to amplify Ehrlichia spp. sodb genes. Primers designed to amplify a 311 bp portion of PME gltA (EPM-gltA-Forward, 5′-CGTGTTTTTCTGCCTTAGCTGCAC and EPM-gltA-Reverse, 5′-CGGCCCAGAAGAACC TGTCA) based on a published PME gltA sequence (DQ363995.1) and a second set of primers designed to target a 480 bp portion of PME map1 (EPM-map1-Forward-3, 5′-CGAGAGCCAACGTTTACAT and EPM-map1-Reverse-2, 5′-GTACCAATACCTGCACATAC) based on published PME map1 sequences (DQ324368.1, EU272373.1 and EU272355.1) were used. Primers designed to amplify a 300 bp portion of sodb (sodb-Forward, 5′-TTTAATAATGCTGGTCAAGTATGGAATCAT and sodb-Reverse, 5′-AAGCGTGTTCCCATACATCCATAG) based on published Ehrlichia spp. sodb sequences (AF392615.1, CP000107.1) were used. Reactions contained 5 μL of DNA extract, 12.5 μL of MyTaqHS-2Xb, 0.125 μM (or 0.25 μM for sodb primers) of each primerc and RNAse-free, molecular-grade water to a final volume of 25 μL. All reactions were performed in a thermocyclerd with an aluminum block under the following conditions, with respective primer annealing temperatures specified: initial temperature at 94°C for 3 minutes, 55 cycles consisting of denaturation at 94°C for 10 seconds, annealing at 68°C (gltA), 62°C (map1), 58°C (sodb) for 10 seconds, extension at 72°C for 15 seconds, and a final extension at 72°C for 30 seconds. Negative controls (RNAse-free, molecular-grade water and uninfected canine genomic DNA) were included in all assays. Amplified PCR products were sequenced directlye and alignments were made with GenBank sequences using AlignX software.f Sequence identities for the partial gltA and map1 genes are as follows, all with 100% coverage: gltA, 100% similar (269 bp) to PME partial gltA gene from an infected goat (DQ363995.1) and Amblyomma americanum tick (EU272374.1); map1, 100% similar (460 bp) to PME partial map1 gene from an infected goat (DQ324368.1) and A. americanum tick (EU272373.1). Before this study, there was no sequence for PME sodb deposited in GenBank. The highest sequence identities assigned by BLASTg for the partial gene amplified using Ehrlichia spp. sodb primers are as follows, all with 100% coverage: sodb, 89% (264/295 bp) similar to multiple strains of E. ruminantium sodb gene, with the highest Max scores reported for Senegal (DQ647026.1), Pokoase (DQ647024.1), and Kumm1 (DQ647023.1) strains, 83% (245/295 bp) similar to E. canis strain Jake sodb (CP000107.1) and 80% (236/295 bp) similar to E. chaffeensis strain Arkansas sodb (AF392615.1). The PME partial sodb gene sequence was submitted to Genbank (Accession number KC702804). Primer sets specific for Rickettsia ompA, E. canis p30, E. ewingii p28, and E. chaffeensis p28 did not amplify DNA from the PME-infected blood sample, suggesting this dog was not likely coinfected with Rickettsia or other Ehrlichia species. Retrospective PCRs using DNA extracted from predoxycycline splenic cytologic smears were negative (16S rRNA, gltA, map1, sodb, E. canis p30, E. ewingii p28, and E. chaffeensis p28) for pathogen DNA. Treatment for suspected lymphoma was not initiated because of the possibility that lymphocytosis and other lymphocytic abnormalities were due to a reactive process secondary to ehrlichiosis.2-4 Treatment with doxycycline (approximately 5 mg/kg PO q12h) was commenced for 30 days. After starting doxycycline treatment, occasional vomiting was noted. Resolution of the thrombocytopenia and lymphocytosis occurred 1 week after starting the treatment. ALP and ALT activities remained increased at 1,989 IU/L and 119 IU/L, respectively. One week after completion of doxycycline, repeat aspiration cytology identified a mixed lymphoid population with expansion of intermediate lymphocytes and mild extramedullary hematopoiesis within the spleen, a mixed lymphoid population within the popliteal lymph node, and an expansion of intermediate lymphocytes and mild vacuolar change within the liver. Flow cytometry of the liver showed a population of large, granular cells with positive intracellular staining for CD3, consistent with T-cell lymphoma. When cytology was repeated again 4 weeks after completion of doxycycline, there was resolution of the abnormal lymphoid population in liver and lymph nodes, but there were increased numbers of lymphoblasts and mild persistent extramedullary hematopoiesis within the spleen (Fig 3). ALP and ALT activities had decreased to 1,352 IU/L and 66 IU/L, respectively. Throughout the 3-month time period described in this case report, the owners reported no clinical abnormalities other than occasional gastrointestinal signs, but felt in retrospect that the dog may have been slightly lethargic as they reported it was more energetic after completion of the doxycycline treatment regimen. Furthermore, 5 subsequent CBCs performed over the next 6 months remained normal. EDTA-anticoagulated whole blood and convalescent serum samples collected from the dog approximately 2 weeks after starting doxycycline treatment were PCR negative for PME (16S rRNA, gltA, map1 and sodb) and the SNAP 4Dx Plus was positive for anti-Ehrlichia spp. antibodies, respectively. Convalescent serum, collected approximately 5 weeks after completion of antibiotic treatment, was minimally reactive (1:32) to R. rickettsii antigens, whereas the dog remained E. canis seroreactive by both IFA (1:512) and SNAP 4Dx Plusa (weak Ehrlichia spp. positive). EDTA whole blood collected at this time remained PCR negative (16S rRNA, gltA, map1 and sodb) for PME. In this report, we provide molecular evidence of PME in a thrombocytopenic dog with abnormal lymphocytosis and clonal T-cell expansion. Treatment with doxycycline resulted in resolution of thrombocytopenia, abnormal lymphocytosis, and abnormal lymphoid cells in liver and lymph nodes, supporting a potential role for PME as a cause of host immune dysregulation. These findings also support a potential pathogenic role for PME as a cause of thrombocytopenia in dogs from the United States. Cytopathology and flow cytometry in this case identified a large number of atypical lymphocytes in the peripheral blood and in hepatic and splenic tissue aspirates with a high number of intermediate lymphocytes, granular CD3+ cells, and clonal T-cell expansion. Intracellular infections that induce cell-mediated immunity can cause cytological changes similar to malignancy.5, 6 A study characterizing peripheral blood smears in human ehrlichiosis patients infected with either E. chaffeensis or E. ewingii documented prominent large granular lymphocytes with atypical, folded, hyperchromatic nuclei that might be confused with neoplastic NK or NK-like T-cells.6 Immunophenotypes compared between dogs with naturally acquired canine monocytic ehrlichiosis (CME) and healthy dogs found that dogs with CME had higher relative numbers of CD3+ T-cells in peripheral blood than did healthy dogs.7 Additional studies found that dogs experimentally infected with E. canis had transitory CD8+ lymphocytosis in both peripheral blood and lymph nodes.8, 9 Additional studies describe clinically ill, E. canis-seropositive dogs with an increasing percentage of peripheral blood CD8+ lymphocytes.3, 10 PARR results from the dog in this case demonstrated clonal T-cell expansion, typically associated with malignancy. At least 2 reports, however, describe this phenomenon in dogs with E. canis infections.2, 4 In a previous report, an E. canis-seropositive dog with pancytopenia and clonal T-cell expansion was treated with doxycycline, which resolved the pancytopenia and the dog remained healthy 2 years later.2 Further investigation is needed to determine if expanded T-cells in E. canis infections are transitory or potentially could develop into lymphoid malignancy, particularly in association with chronic undiagnosed infections. As current evidence suggests, a role for Ehrlichia spp. in immune dysregulation is likely, but the extent may be subject to host factors, species and strain virulence, as well as phase of the disease. CME has acute, subclinical, and chronic phases representing different infection durations and variations in clinical disease manifestations. One recent study, however, found no statistically significant differences among CD3+, CD8+, and, CD4+ cells in peripheral blood samples from dogs with clinical or subclinical CME.11 Signs of lymphoid malignancy continue to be monitored in the dog of this report. Resolution of the immunologic abnormalities noted in liver, lymph node, and blood after doxycycline treatment, however, strongly supports an infectious etiology. To the authors’ knowledge, this is the first report of PME infection in a dog. Genetically and antigenically similar to E. ruminantium, PME was first identified by PCR in a goat experimentally infested with A. americanum ticks collected from Panola Mountain State Park, Georgia.12 Goats infected with PME developed serous nasal discharge and febrile illness with hematologic changes consisting of decreased ALP activities and neutropenia; rare morulae in mononuclear cells were identified in 1 goat.12, 13 PME also was detected by PCR in whole blood from a man in Atlanta, GA who developed myalgia after being bitten by a nymphal A. americanum tick.14 No hematologic abnormalities were reported, and clinical signs resolved after doxycycline treatment. The role, if any, of PME in the overall pathogenesis of the various disease manifestations described in the dog of this report over its 3 year history of illness is impossible to assess. At the time PME was identified, the dog was asymptomatic and hematological abnormalities were limited to thrombocytopenia, increased numbers of atypical lymphocytes, and progressively increasing ALP activity (after initiation of doxycycline). The most notable characteristics of illness that potentially were related to PME infection in this dog included thrombocytopenia, cytological changes in the liver, lymph node and spleen, and clonal T-cell expansion. After treatment with doxycycline, sequential resolution of most of these abnormalities occurred. It is not clear when the dog became infected with PME, but serum screened for exposure to vector-borne pathogens 8 months earlier was negative, and the dog's owners reported flea and tick exposure over the previous 6-month time period. PME exposure presumably occurred in central NC because the dog had no travel history outside of the state. This is not a surprising finding, given documentation of PME in A. americanum from the eastern United States, with PME-positive ticks detected in FL, GA, KY, NJ, and NY.15 In addition, deer from AR, NC, and VA were PCR positive for PME and were shown to be competent reservoirs for the pathogen.16 Documentation of PME in this dog supports the possibility that this tick-borne organism may represent an unrecognized human or ruminant pathogen in NC and surrounding states. Currently, serological diagnostics specific for PME are not available. Serum from goats infected with PME was weakly IFA seropositive to E. chaffeensis and seropositive by ELISA to the MAP1 protein from E. ruminantium.12, 13 Serum from the dog of this report reacted strongly with E. canis antigen by IFA, but less strongly with the synthetic antigens used in a commercial ELISA (SNAP 4Dx Plus). Antibodies to other Ehrlichia spp. such as E. chaffeensis have been shown to cross-react with E. canis antigens, and it is likely that the reactivity from this PME-infected dog also represented a cross-reaction.17 Whereas E. canis, E. ewingii, and E. chaffeensis were not detected by PCR, we cannot rule out the possibility that seroreactivity was because of previous exposure with one of these pathogens. In addition to highlighting the emergence of vector-borne pathogens in a novel host species, the findings in this case report underscore the challenges faced in diagnosing canine lymphocytic malignancies and reinforce the need to better understand the immunopathology of canine ehrlichiosis, which can be caused by E. canis, E. chaffeensis, E. ewingii, E. muris, and PME in dogs in North America. Documentation of abnormal or expanded lymphocyte populations in the blood or tissues of dogs should prompt diagnostic consideration of infection with an intracellular pathogen, including Ehrlichia spp. Conflict of Interest: Authors disclose no conflict of interest.}, number={5}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Qurollo, B.A. and Davenport, A.C. and Sherbert, B.M. and Grindem, C.B. and Birkenheuer, A.J. and Breitschwerdt, E.B.}, year={2013}, month={Jul}, pages={1251–1255} }
 @article{chaston_suen_tucker_andersen_bhasin_bode_bode_brachmann_cowles_cowles_et al._2011, title={The Entomopathogenic Bacterial Endosymbionts Xenorhabdus and Photorhabdus: Convergent Lifestyles from Divergent Genomes}, volume={6}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0027909}, DOI={10.1371/journal.pone.0027909}, abstractNote={Members of the genus Xenorhabdus are entomopathogenic bacteria that associate with nematodes. The nematode-bacteria pair infects and kills insects, with both partners contributing to insect pathogenesis and the bacteria providing nutrition to the nematode from available insect-derived nutrients. The nematode provides the bacteria with protection from predators, access to nutrients, and a mechanism of dispersal. Members of the bacterial genus Photorhabdus also associate with nematodes to kill insects, and both genera of bacteria provide similar services to their different nematode hosts through unique physiological and metabolic mechanisms. We posited that these differences would be reflected in their respective genomes. To test this, we sequenced to completion the genomes of Xenorhabdus nematophila ATCC 19061 and Xenorhabdus bovienii SS-2004. As expected, both Xenorhabdus genomes encode many anti-insecticidal compounds, commensurate with their entomopathogenic lifestyle. Despite the similarities in lifestyle between Xenorhabdus and Photorhabdus bacteria, a comparative analysis of the Xenorhabdus, Photorhabdus luminescens, and P. asymbiotica genomes suggests genomic divergence. These findings indicate that evolutionary changes shaped by symbiotic interactions can follow different routes to achieve similar end points.}, number={11}, journal={PLoS ONE}, publisher={Public Library of Science (PLoS)}, author={Chaston, John M. and Suen, Garret and Tucker, Sarah L. and Andersen, Aaron W. and Bhasin, Archna and Bode, Edna and Bode, Helge B. and Brachmann, Alexander O. and Cowles, Charles E. and Cowles, Kimberly N. and et al.}, editor={Badger, Jonathan H.Editor}, year={2011}, month={Nov}, pages={e27909} }
 @article{kennedy_qurollo_rose_thamm_2010, title={Epidermal growth factor enhances the malignant phenotype in canine mammary carcinoma cell lines}, volume={9}, ISSN={1476-5810}, url={http://dx.doi.org/10.1111/j.1476-5829.2010.00248.x}, DOI={10.1111/j.1476-5829.2010.00248.x}, abstractNote={Canine mammary gland tumours (CMTs) are the most common malignancies in female dogs. The receptor tyrosine kinase EGFR (erbb1), a receptor for epidermal growth factor (EGF) and related factors, mediates multiple oncogenic functions in human epithelial neoplasms. While previous studies have demonstrated EGFR expression in canine tumours, its function has not been studied in canine cancer. The purpose of this study was to determine the in vitro effects of EGF and vandetanib (ZD6474), a small molecule inhibitor of VEGFR‐2, EGFR and RET tyrosine kinases, on proliferation, invasion, survival and chemosensitivity in CMT cells. In low serum, EGF enhanced proliferation and chemotaxis, attenuated apoptosis, and stimulated vascular endothelial growth factor (VEGF) production. Vandetanib dose‐dependently inhibited EGFR phosphorylation as well as PI3K/Akt activation, and inhibited all EGF‐induced phenotypic effects. In conclusion, EGF stimulates multiple features promoting the malignant phenotype in CMT. Thus, CMT may be an important translational model for the investigation of novel EGFR‐directed therapies.}, number={3}, journal={Veterinary and Comparative Oncology}, publisher={Wiley}, author={Kennedy, K. C. and Qurollo, B. A. and Rose, B. J. and Thamm, D. H.}, year={2010}, month={Nov}, pages={196–206} }
 @article{thamm_huelsmeyer_mitzey_qurollo_rose_kurzman_2010, title={RT-PCR-based tyrosine kinase display profiling of canine melanoma: IGF-1 receptor as a potential therapeutic target}, volume={20}, ISSN={0960-8931}, url={http://dx.doi.org/10.1097/cmr.0b013e328331ca86}, DOI={10.1097/cmr.0b013e328331ca86}, abstractNote={Canine malignant melanoma (CMM) resembles human malignant melanoma in terms of metastatic behavior, refractoriness to standard therapy, and tumor antigen expression but it is largely unknown how CMM resembles human melanoma with regard to molecular pathogenesis and cellular signaling. No attempt has been made to systematically define the repertoire of tyrosine kinases (TKs) expressed in CMM. This study used a reverse transcription-PCR display technique to evaluate the expression of multiple TKs in the 17CM98 CMM cell line. RT-PCR was performed using degenerate primers coding for highly conserved regions flanking the kinase domains of many TKs and the repertoire of TKs expressed was determined using standard molecular cloning techniques. Sequencing 46 clones yielded canine homologs of insulin-like growth factor-1 receptor (IGF-1R) (50%), JAK1 (17%), PDGFR-a (11%), FGFR1 (9%), Axl (7%), Abl (4%), and PTK2 (2%). Interestingly, IGF-1R, JAK1, and Axl were detected in human melanoma using similar techniques, supporting the cross-species validity of this assay. Given the abundance of IGF-1R clones, we determined the biological effect of rhIGF-1 in 17CM98 cells. IGF-1 stimulated cell proliferation and vascular endothelial growth factor production in 17CM98, and addition of the IGF-1R inhibitor ADW742 abrogated IGF-1-induced phenotypic changes. Expression of IGF-1R mRNA was detected in five of five additional CMM cell cultures, and IGF-1R protein was detected in five of six primary tumors evaluated, suggesting that IGF-1R expression may be common in CMM and may provide a novel target for future therapy. In conclusion, this study suggests that similar TKs are expressed in human and canine melanoma, and shows potential antitumor effects of IGF-1R inhibition in CMM.}, number={1}, journal={Melanoma Research}, publisher={Ovid Technologies (Wolters Kluwer Health)}, author={Thamm, Douglas H. and Huelsmeyer, Michael K. and Mitzey, Ann M. and Qurollo, Barbara and Rose, Barbara J. and Kurzman, Ilene D.}, year={2010}, month={Feb}, pages={35–42} }
 @article{davis_simpson_nugent_carroll_avery_rali_chen_qurallo_quinn_2008, title={Pim2 Inhibitors from the Papua New Guinean PlantCupaniopsis macropetala⊥}, volume={71}, ISSN={0163-3864 1520-6025}, url={http://dx.doi.org/10.1021/np070431w}, DOI={10.1021/np070431w}, abstractNote={Bioassay-guided fractionation of an organic extract from the leaves of Cupaniopsis macropetala resulted in the isolation of a new alkaloid, galloyl tyramine ( 1), together with the known flavonoid glycoside quercitrin ( 2). The structure of 1 was determined following 1D and 2D NMR, IR, UV, and MS data analysis. Compounds 1 and 2 displayed IC 50 values of 161 and 25 microM, respectively, in a Pim2 enzyme assay.}, number={3}, journal={Journal of Natural Products}, publisher={American Chemical Society (ACS)}, author={Davis, Rohan A. and Simpson, Moana M. and Nugent, Ryan B. and Carroll, Anthony R. and Avery, Vicky M. and Rali, Topul and Chen, Huawei and Qurallo, Barbara and Quinn, Ronald J.}, year={2008}, month={Mar}, pages={451–452} }
 @article{qurollo_bishop_hassan_2001, title={Characterization of the iron superoxide dismutase gene of <i>Azotobacter vinelandii</i>: <i>sodB</i> may be essential for viability}, volume={47}, ISSN={1480-3275 0008-4166}, url={http://dx.doi.org/10.1139/cjm-47-1-63}, DOI={10.1139/cjm-47-1-63}, number={1}, journal={Canadian Journal of Microbiology}, publisher={Canadian Science Publishing}, author={Qurollo, Barbara A. and Bishop, Paul E. and Hassan, Hosni M.}, year={2001}, pages={63–71} }
 @article{goodner_hinkle_gattung_miller_blanchard_qurollo_goldman_cao_askenazi_halling_et al._2001, place={Science (New York, N.Y}, title={Genome Sequence of the Plant Pathogen and Biotechnology Agent Agrobacterium tumefaciens C58}, volume={294}, ISSN={0036-8075 1095-9203}, url={http://dx.doi.org/10.1126/science.1066803}, DOI={10.1126/science.1066803}, abstractNote={
            Agrobacterium tumefaciens
            is a plant pathogen capable of transferring a defined segment of DNA to a host plant, generating a gall tumor. Replacing the transferred tumor-inducing genes with exogenous DNA allows the introduction of any desired gene into the plant. Thus,
            A. tumefaciens
            has been critical for the development of modern plant genetics and agricultural biotechnology. Here we describe the genome of
            A. tumefaciens
            strain C58, which has an unusual structure consisting of one circular and one linear chromosome. We discuss genome architecture and evolution and additional genes potentially involved in virulence and metabolic parasitism of host plants.
          }, number={5550}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Goodner, B. and Hinkle, G. and Gattung, S. and Miller, N. and Blanchard, M. and Qurollo, B. and Goldman, B.S. and Cao, Y. and Askenazi, M. and Halling, C. and et al.}, year={2001}, month={Dec}, pages={2323–2328} }