@article{fingerhood_neupane_breitschwerdt_choi_2024, title={Diagnostic challenge in veterinary pathology: Tri-cavitary effusion in a cat with systemic pyogranulomatous inflammation}, volume={1}, ISSN={["1544-2217"]}, DOI={10.1177/03009858241226648}, abstractNote={An approximately 2-month-old, domestic shorthair cat presented to the University of California, Davis Veterinary Medical Teaching Hospital for lethargy, hyporexia, and increased respiratory rate and effort of a few days duration.Clinically, there was tri-cavitary effusion; the pleural effusion, interpreted as a borderline modified transudate, had a nucleated cell count of 130/µl (43% neutrophils, 29% small mononuclear cells, 28% large mononuclear cells), a total protein of 2.5 g/dl, and frequent erythrophagia.Complete blood count abnormalities included an inflammatory leukogram with a left shift (4680 band/µl, 120 metamyelocytes/µl), toxic neutrophils, and a nonregenerative anemia (17.2% hematocrit [30%-50%]).Serum biochemical abnormalities included hypoglobulinemia (2.0 g/dl [2.8-5.4]) with a low-normal albumin (2.4 g/dl [2.2-4.6]),hyponatremia (133 mmol/l [151-158]), hypokalemia (3.1 mmol/l [3.6-4.9]),hypochloremia (101 mmol/l [117-126]), hypophosphatemia (3 mg/dl [3.2-6.3]), and hypoglycemia (59 mg/dl [63-118]).Pleural fluid was quantitative PCR negative for feline coronavirus (feline infectious peritonitis [FIP]), and a feline leukemia virus antigen SNAP test (IDEXX Laboratories, Westbrook, ME, USA) was negative.Echocardiography and chest radiographs did not identify congenital cardiac abnormalities.Due to a guarded prognosis, humane euthanasia was elected.}, journal={VETERINARY PATHOLOGY}, author={Fingerhood, Sai and Neupane, Pradeep and Breitschwerdt, Edward B. and Choi, Eunju April}, year={2024}, month={Jan} }
 @article{moore_lashnits_neupane_herrin_lappin_andre_breitschwerdt_2023, title={Feeding on a Bartonella henselae Infected Host Triggers Temporary Changes in the Ctenocephalides felis Microbiome}, volume={12}, ISSN={["2076-0817"]}, url={https://doi.org/10.3390/pathogens12030366}, DOI={10.3390/pathogens12030366}, abstractNote={The effect of Bartonella henselae on the microbiome of its vector, Ctenocephalides felis (the cat flea) is largely unknown, as the majority of C. felis microbiome studies have utilized wild-caught pooled fleas. We surveyed the microbiome of laboratory-origin C. felis fed on B. henselae-infected cats for 24 h or 9 days to identify changes to microbiome diversity and microbe prevalence compared to unfed fleas, and fleas fed on uninfected cats. Utilizing Next Generation Sequencing (NGS) on the Illumina platform, we documented an increase in microbial diversity in C. felis fed on Bartonella-infected cats for 24 h. These changes returned to baseline (unfed fleas or fleas fed on uninfected cats) after 9 days on the host. Increased diversity in the C. felis microbiome when fed on B. henselae-infected cats may be related to the mammalian, flea, or endosymbiont response. Poor B. henselae acquisition was documented with only one of four infected flea pools having B. henselae detected by NGS. We hypothesize this is due to the use of adult fleas, flea genetic variation, or lack of co-feeding with B. henselae-infected fleas. Future studies are necessary to fully characterize the effect of endosymbionts and C. felis diversity on B. henselae acquisition.}, number={3}, journal={PATHOGENS}, author={Moore, Charlotte and Lashnits, Erin and Neupane, Pradeep and Herrin, Brian H. H. and Lappin, Michael and Andre, Marcos Rogerio and Breitschwerdt, Edward B. B.}, year={2023}, month={Mar} }
 @article{neupane_maggi_basnet_lashnits_andrews_breitschwerdt_2022, title={Bartonella henselae Recombinant Pap31 for the Diagnosis of Canine and Human Bartonelloses}, volume={11}, ISSN={["2076-0817"]}, url={https://www.mdpi.com/2076-0817/11/2/182}, DOI={10.3390/pathogens11020182}, abstractNote={Bartonella spp. comprise a genus of Gram-negative alphaproteobacteria that are slow growing, fastidious, and facultative intracellular pathogens with zoonotic potential. Immunofluorescent antibody assays (IFAs), Western blot (WB), and enzyme-linked immunosorbent assays (ELISAs), the frequently used modalities for the serological diagnosis of canine and human Bartonelloses, generate numerous false negative results. Therefore, the development of a reliable serodiagnostic assay for Bartonelloses is of clinical and epidemiological importance. Pap31, a heme binding surface protein of B. henselae, is associated with bacterial adhesion and related to bacterial colonization. To our knowledge, B. henselae Pap31 and its fragments (N-terminal (NTD), middle (MD), and C-terminal (CTD) domains) have not been investigated for the serodiagnosis of canine and human Bartonelloses. In this study, we evaluate the diagnostic utility of B. henselae recombinant whole Pap31 (rPap31) and Pap31 fragments by ELISA using sera from 70 dogs (36 Bartonella spp. IFA-positive (naturally infected), and 34 Bartonella spp. IFA- and PCR-negative (control dogs)) and 36 humans (18 Bartonella spp. IFA-positive (naturally infected) and 18 controls)). In the dogs, the area under the curve (AUC) score of recombinant whole Pap31 was 0.714 with a sensitivity of 42% and specificity of 94% at an OD cutoff value of 0.8955. Among the evaluated recombinant Pap31 proteins for the diagnosis of canine Bartonelloses, rPap31-NTD yielded the highest AUC score of 0.792 (95% CI 0.688–0.895) with a sensitivity of 44% and specificity of 100% at a cutoff value of 1.198. In concordance with this finding, rPap31-NTD also had the highest AUC score of 0.747 (95% CI 0.581–0.913) among the Pap31 recombinant proteins for the diagnosis of human Bartonelloses, with 39% sensitivity and 94% specificity at a cutoff value of 1.366. Recombinant whole Pap31 (rPap31) resulted in 72% sensitivity and 61% specificity at a cutoff value of 0.215 for human Bartonelloses. Due to either low sensitivity or questionable specificity, our findings indicate that recombinant Pap31 and the selected fragments may not be appropriate diagnostic targets in detecting anti-Bartonella antibodies in Bartonella-infected dogs and humans. The findings from this study can be used to further assess the antigenicity and immunogenicity of B. henselae Pap31 as a diagnostic target.}, number={2}, journal={PATHOGENS}, publisher={MDPI AG}, author={Neupane, Pradeep and Maggi, Ricardo G. and Basnet, Manoj and Lashnits, Erin and Andrews, Gerard P. and Breitschwerdt, Edward B.}, year={2022}, month={Feb} }
 @article{andre_neupane_lappin_herrin_smith_williams_collins_bai_jorge_balbuena_et al._2022, title={Using Proteomic Approaches to Unravel the Response of Ctenocephalides felis felis to Blood Feeding and Infection With Bartonella henselae}, volume={12}, ISSN={["2235-2988"]}, url={http://dx.doi.org/10.3389/fcimb.2022.828082}, DOI={10.3389/fcimb.2022.828082}, abstractNote={Among the Ctenocephalides felis felis-borne pathogens, Bartonella henselae, the main aetiological agent of cat scratch disease (CSD), is of increasing comparative biomedical importance. Despite the importance of B. henselae as an emergent pathogen, prevention of the diseases caused by this agent in cats, dogs and humans mostly relies on the use of ectoparasiticides. A vaccine targeting both flea fitness and pathogen competence is an attractive choice requiring the identification of flea proteins/metabolites with a dual effect. Even though recent developments in vector and pathogen -omics have advanced the understanding of the genetic factors and molecular pathways involved at the tick-pathogen interface, leading to discovery of candidate protective antigens, only a few studies have focused on the interaction between fleas and flea-borne pathogens. Taking into account the period of time needed for B. henselae replication in flea digestive tract, the present study investigated flea-differentially abundant proteins (FDAP) in unfed fleas, fleas fed on uninfected cats, and fleas fed on B. henselae-infected cats at 24 hours and 9 days after the beginning of blood feeding. Proteomics approaches were designed and implemented to interrogate differentially expressed proteins, so as to gain a better understanding of proteomic changes associated with the initial B. henselae transmission period (24 hour timepoint) and a subsequent time point 9 days after blood ingestion and flea infection. As a result, serine proteases, ribosomal proteins, proteasome subunit α-type, juvenile hormone epoxide hydrolase 1, vitellogenin C, allantoinase, phosphoenolpyruvate carboxykinase, succinic semialdehyde dehydrogenase, glycinamide ribotide transformylase, secreted salivary acid phosphatase had high abundance in response of C. felis blood feeding and/or infection by B. henselae. In contrast, high abundance of serpin-1, arginine kinase, ribosomal proteins, peritrophin-like protein, and FS-H/FSI antigen family member 3 was strongly associated with unfed cat fleas. Findings from this study provide insights into proteomic response of cat fleas to B. henselae infected and uninfected blood meal, as well as C. felis response to invading B. henselae over an infection time course, thus helping understand the complex interactions between cat fleas and B. henselae at protein levels.}, journal={FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY}, publisher={Frontiers Media SA}, author={Andre, Marcos Rogerio and Neupane, Pradeep and Lappin, Michael and Herrin, Brian and Smith, Vicki and Williams, Taufika Islam and Collins, Leonard and Bai, Hongxia and Jorge, Gabriel Lemes and Balbuena, Tiago Santana and et al.}, year={2022}, month={Jan} }
 @article{lashnits_thatcher_carruth_mestek_buch_beall_neupane_chandrashekar_breitschwerdt_2021, title={Bartonella spp. seroepidemiology and associations with clinicopathologic findings in dogs in the United States}, volume={36}, ISSN={["1939-1676"]}, url={https://doi.org/10.1111/jvim.16311}, DOI={10.1111/jvim.16311}, abstractNote={AbstractBackgroundImproved understanding of Bartonella spp. serology in dogs may aid clinical decision making.ObjectiveDescribe demographic and geographic patterns of Bartonella spp. seroreactivity in dogs, and describe hematologic and serum biochemical abnormalities in Bartonella spp. seroreactive and nonseroreactive dogs.AnimalsSerum samples from 5957 dogs in the United States, previously submitted to IDEXX Reference Laboratories.MethodsSerum was tested using 3 indirect ELISAs for B. henselae, B. vinsonii subsp. berkhoffii, and B. koehlerae. Complete blood count and serum biochemistry panel results were reviewed retrospectively.ResultsOverall, 6.1% of dogs were Bartonella spp. seroreactive. Toy breeds were less likely to be seroreactive (3.9%) than mixed breeds (7.5%; adjusted odds ratio [aOR], 0.48; 95% confidence interval [CI], 0.32‐0.72), and dogs <1 year old were less likely to be seroreactive (3.4%) than dogs 1 to 5.5 years of age (7.3%; aOR, 0.42; 95% CI, 0.23‐0.72). Dogs in the West South Central (9.8%) and South Atlantic (8.8%) regions were more likely than dogs elsewhere in the United States to be seroreactive (aOR, 2.22; 95% CI, 1.31‐3.87; aOR, 2.44; 95% CI, 1.38‐4.36).Conclusions and Clinical ImportanceDemographic and geographic findings for Bartonella spp. exposure were broadly comparable to previously reported patterns.}, number={1}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, publisher={Wiley}, author={Lashnits, Erin and Thatcher, Brendon and Carruth, Ariel and Mestek, Anton and Buch, Jesse and Beall, Melissa and Neupane, Pradeep and Chandrashekar, Ramaswamy and Breitschwerdt, Edward B.}, year={2021}, month={Nov} }
 @article{lashnits_neupane_bradley_richardson_maggi_breitschwerdt_2021, title={Comparison of Serological and Molecular Assays for Bartonella Species in Dogs with Hemangiosarcoma}, volume={10}, ISSN={["2076-0817"]}, url={https://doi.org/10.3390/pathogens10070794}, DOI={10.3390/pathogens10070794}, abstractNote={Currently, a gold standard diagnostic test for Bartonella infection in dogs is lacking. This represents a critical limitation for the development and evaluation of new diagnostic tests, as well as for the diagnosis of, and research on, bartonellosis in dogs. This retrospective observational study aims to compare the results of commonly performed and newly-reported Bartonella spp. diagnostic tests in banked clinical specimens from 90 dogs with hemangiosarcoma (HSA) using composite reference standard (CRS) and random effects latent class analysis (RE-LCA) techniques. Samples from each dog were tested using six serological or molecular diagnostic assays, including indirect fluorescent antibody (IFA) and Western blot (WB) for the detection of antibodies in serum, and qPCR and droplet digital PCR (ddPCR) in blood and fresh frozen tissue biopsy samples (mainly splenic HSA tumors and histopathologically normal spleen or skin/adipose tissue). Bartonella infection prevalence was estimated to be 78% based on the CRS (parallel testing with all six assays), and 64% based on the RE-LCA model. The assay with the highest diagnostic accuracy was qPCR performed on fresh frozen tissue biopsy samples (sensitivity: 94% by RE-LCA and 80% by CRS; specificity: 100%). When comparing newly-reported to traditional Bartonella diagnostic assays, ddPCR was more sensitive for the detection of Bartonella DNA than qPCR when testing blood samples (36% vs. 0%, p < 0.0001). Dogs that were positive on serological assays alone with negative molecular assays were highly unlikely (<3%) to be classified as infected by the RE-LCA model. These data indicate that Bartonella spp. DNA can be PCR amplified from fresh frozen tissues from a majority of dogs with HSA using both qPCR and ddPCR, supporting the use of these methods for future controlled studies comparing the prevalence of Bartonella spp. DNA in the tissue of dogs with HSA to that of unaffected controls.}, number={7}, journal={PATHOGENS}, publisher={MDPI AG}, author={Lashnits, Erin and Neupane, Pradeep and Bradley, Julie M. and Richardson, Toni and Maggi, Ricardo G. and Breitschwerdt, Edward B.}, year={2021}, month={Jul} }
 @article{neupane_sevala_balakrishnan_marr_wilson_maggi_birkenheuer_lappin_chomel_breitschwerdt_2020, title={Validation of Bartonella henselae Western Immunoblotting for Serodiagnosis of Bartonelloses in Dogs}, volume={58}, ISSN={["1098-660X"]}, DOI={10.1128/JCM.01335-19}, abstractNote={
            Bartonella
            spp. are etiological agents of life-threatening zoonotic diseases in dogs worldwide. Due to the poor sensitivity of immunofluorescent-antibody assays (IFAs), a reliable serodiagnostic test for canine bartonelloses is of clinical importance. The utility of Western blotting (WB) for the serodiagnosis of canine bartonelloses has not been critically investigated. The objective of this study was to characterize WB immunodominant proteins that could be used to confirm a serodiagnosis of bartonelloses.
          }, number={4}, journal={JOURNAL OF CLINICAL MICROBIOLOGY}, author={Neupane, Pradeep and Sevala, Sindhura and Balakrishnan, Nandhakumar and Marr, Henry and Wilson, James and Maggi, Ricardo and Birkenheuer, Adam and Lappin, Michael and Chomel, Bruno and Breitschwerdt, Edward B.}, year={2020}, month={Apr} }
 @article{southern_neupane_ericson_dencklau_linder_bradley_mckeon_long_breitschwerdt_2018, title={Bartonella henselae
 in a dog with ear tip vasculitis}, volume={29}, ISSN={0959-4493}, url={http://dx.doi.org/10.1111/vde.12695}, DOI={10.1111/vde.12695}, abstractNote={BackgroundBartonella henselae, a Gram‐negative, zoonotic, alpha‐proteobacteria has been previously implicated in association with cutaneous vasoproliferative lesions (bacillary angiomatosis), nodular panniculitis and multifocal erythema (erythema multiforme) in dogs.ObjectiveDescribe clinical, microbiological and histological lesions in a dog with ear margin vasculitis and B. henselae infection.AnimalsA 12‐month‐old, specific pathogen‐free intact female beagle dog maintained in a vector‐free laboratory animal resource facility.Methods and materialsBartonella and Rickettsia serological evaluation, Bartonella and Rickettsia PCR, Bartonella alpha‐proteobacteria growth medium (BAPGM) enrichment blood culture/PCR, histopathological investigation and confocal immunohistochemical evaluation.ResultsSerological investigation (seroreversion) and PCR testing of aural tissue biopsies failed to support Rickettsia rickettsii as a cause of the aural vasculitis; however, B. henselae, genotype San Antonio 2 DNA was amplified and sequenced from both ear tip margins and from normal‐appearing abdominal skin. Seroconversion to B. henselae was documented retrospectively by IFA testing. Bartonella henselae organisms were visualized by confocal immunostaining within all three biopsies. Histopathology revealed small vessel necrotizing vasculitis and dermal necrosis. Bartonella henselae seroreversion and complete resolution of skin lesions occurred in conjunction with administration of oral doxycycline and enrofloxacin for six weeks.Conclusions and Clinical ImportanceBartonella henselae is an emerging zoonotic pathogen that has been associated with leucocytoclastic vasculitis in humans and may have had a contributing or causative role in the development of the cutaneous aural margin vasculitis in this beagle.}, number={6}, journal={Veterinary Dermatology}, publisher={Wiley}, author={Southern, Brittany L. and Neupane, Pradeep and Ericson, Marna E. and Dencklau, Jamie C. and Linder, Keith E. and Bradley, Julie M. and McKeon, Gabriel P. and Long, Charles T. and Breitschwerdt, Edward B.}, year={2018}, month={Oct}, pages={537–e180} }
 @article{neupane_hegarty_marr_maggi_birkenheuer_breitschwerdt_2018, title={Evaluation of cell culture-grown Bartonella
 antigens in immunofluorescent antibody assays for the serological diagnosis of bartonellosis in dogs}, volume={32}, ISSN={0891-6640}, url={http://dx.doi.org/10.1111/jvim.15301}, DOI={10.1111/jvim.15301}, abstractNote={BackgroundBecause of poor sensitivity and questionable specificity of immunofluorescent antibody assays (IFAs), serological diagnosis of Bartonella species infections in dogs remains challenging. Despite limitations, IFA testing is the historical “gold standard” for Bartonella serodiagnosis in animals and humans. Because most diagnostic laboratories test against only 1 or 2 Bartonella spp., testing against a broader panel of Bartonella antigens may enhance diagnostic sensitivity and specificity.ObjectiveTo evaluate the sensitivity and specificity of Bartonella IFA using 8 cell culture‐grown Bartonella spp. isolates.AnimalsArchived serum samples from 34 Bartonella spp. naturally exposed, polymerase chain reaction (PCR)‐positive dogs and from 26 PCR‐negative and IFA‐negative dogs.Methods
Bartonella IFA sensitivity and specificity were assessed using cell culture‐grown whole cell antigens derived from 3 Bartonella henselae (Bh) strains (Bh Houston 1, Bh San Antonio Type 2, Bh California 1), 3 Bartonella vinsonii subsp. berkhoffii genotypes (Bvb I, II, and III), Bartonella koehlerae (Bk), and Bartonella quintana (Bq).ResultsOnly 62% of 34 Bartonella spp. PCR‐positive dogs were seroreactive to any of the 8 Bartonella IFA antigens, indicating low IFA sensitivity. PCR‐positive dogs were most often IFA seroreactive to Bq (n = 15), to Bvb II (n = 13), or to both (n = 9) antigens. Of the 26 previously IFA‐negative/PCR‐negative dogs, 4 (15%) were seroreactive using the expanded antigen panel.Conclusion and Clinical ImportanceDespite IFA testing of dogs against 8 different Bartonella isolates, IFA sensitivity remained poor, and specificity was only 85%. Development of a reliable serological assay is needed to facilitate the diagnosis of Bartonella infection in dogs.}, number={6}, journal={Journal of Veterinary Internal Medicine}, publisher={Wiley}, author={Neupane, Pradeep and Hegarty, Barbara C. and Marr, Henry S. and Maggi, Ricardo G. and Birkenheuer, Adam J. and Breitschwerdt, Edward B.}, year={2018}, month={Oct}, pages={1958–1964} }
 @article{kern_swartley_neupane_balakrishnan_breitschwerdt_2019, title={Pasteurella canis infective endocarditis in a dog}, volume={229}, ISSN={0378-1135}, url={http://dx.doi.org/10.1016/j.vetmic.2018.12.001}, DOI={10.1016/j.vetmic.2018.12.001}, abstractNote={Infective endocarditis, an infrequent clinical syndrome in dogs, is typically associated with nondescript clinical signs such as fever, malaise and loss of appetite. Although an uncommonly reported infection in dogs, Pasteurella canis is an emerging pathogen with increasing relevance in the human microbiology literature. The goal of this study is to detail the clinical presentation and microbiological findings associated with a novel causative agent of infective endocarditis in the dog. Diagnostic evaluation as well as conventional, automated and molecular microbiological methods are highlighted. The recent literature regarding P. canis and infective endocarditis in companion animals and humans is reviewed. Although an unusual etiologic agent of infective endocarditis, awareness of P. canis as a diagnostic possibility is crucial to accurate microbial surveillance.}, journal={Veterinary Microbiology}, publisher={Elsevier BV}, author={Kern, Zachary T. and Swartley, Olivia M. and Neupane, Pradeep and Balakrishnan, Nandhakumar and Breitschwerdt, Edward B.}, year={2019}, month={Feb}, pages={14–19} }