@article{lewbart_griffioen_savo_pablo munoz-perez_ortega_loyola_roberts_schaaf_steinberg_osegueda_et al._2018, title={Biochemistry and hematology parameters of the San Cristobal Galapagos tortoise (Chelonoidis chathamensis)}, volume={6}, ISSN={["2051-1434"]}, DOI={10.1093/conphys/coy004}, abstractNote={As part of a planned introduction of captive Galapagos tortoises (Chelonoidis chathamensis) to the San Cristóbal highland farms, our veterinary team performed thorough physical examinations and health assessments of 32 tortoises. Blood samples were collected for packed cell volume (PCV), total solids (TS), white blood cell count (WBC) differential, estimated WBC and a biochemistry panel including lactate. In some cases not all of the values were obtainable but most of the tortoises have full complements of results. Despite a small number of minor abnormalities this was a healthy group of mixed age and sex tortoises that had been maintained with appropriate husbandry. This work establishes part of a scientific and technical database to provide qualitative and quantitative information when establishing sustainable development strategies aimed at the conservation of Galapagos tortoises.}, journal={CONSERVATION PHYSIOLOGY}, author={Lewbart, Gregory A. and Griffioen, John A. and Savo, Alison and Pablo Munoz-Perez, Juan and Ortega, Carlos and Loyola, Andrea and Roberts, Sarah and Schaaf, George and Steinberg, David and Osegueda, Steven B. and et al.}, year={2018}, month={Feb} }
 @article{schreeg_marr_tarigo_sherrill_outi_scholl_bird_vigil_hung_nakajima_et al._2018, title={Identification of Cytauxzoon felis antigens via protein microarray and assessment of expression library immunization against cytauxzoonosis}, volume={15}, ISSN={["1559-0275"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-85059281263&partnerID=MN8TOARS}, DOI={10.1186/s12014-018-9218-9}, abstractNote={Cytauxzoonosis is a disease of felids in North America caused by the tick-transmitted apicomplexan parasite Cytauxzoon felis. Cytauxzoonosis is particularly virulent for domestic cats, but no vaccine currently exists. The parasite cannot be cultivated in vitro, presenting a significant limitation for vaccine development. Recent sequencing of the C. felis genome has identified over 4300 putative protein-encoding genes. From this pool we constructed a protein microarray containing 673 putative C. felis proteins. This microarray was probed with sera from C. felis-infected and naïve cats to identify differentially reactive antigens which were incorporated into two expression library vaccines, one polyvalent and one monovalent. We assessed the efficacy of these vaccines to prevent of infection and/or disease in a tick-challenge model. Probing of the protein microarray resulted in identification of 30 differentially reactive C. felis antigens that were incorporated into the two expression library vaccines. However, expression library immunization failed to prevent infection or disease in cats challenged with C. felis. Protein microarray facilitated high-throughput identification of novel antigens, substantially increasing the pool of characterized C. felis antigens. These antigens should be considered for development of C. felis vaccines, diagnostics, and therapeutics.}, number={1}, journal={CLINICAL PROTEOMICS}, author={Schreeg, Megan E. and Marr, Henry S. and Tarigo, Jaime L. and Sherrill, Meredith K. and Outi, Hilton K. and Scholl, Elizabeth H. and Bird, David M. and Vigil, Adam and Hung, Chris and Nakajima, Rie and et al.}, year={2018}, month={Dec} }
 @misc{gookin_hanrahan_levy_2017, title={The conundrum of feline trichomonosis: The more we learn the "trickier' it gets}, volume={19}, ISSN={["1532-2750"]}, DOI={10.1177/1098612x17693499}, abstractNote={Practical relevance: Trichomonosis of the large intestine of the cat was described as a cause of chronic diarrhea over 20 years ago. The trichomonad was identified as Tritrichomonas foetus, with a genotype that is distinct from venereal T foetus of cattle. Clinical challenges: Despite multiple means for diagnosis of the infection, including light microscopy, protozoal culture and PCR amplification using species-specific primers, tests with even greater sensitivity are needed. Feline trichomonosis is resistant to all commonly used antiprotozoal drugs. Ronidazole is currently the only drug demonstrated to be effective in eliminating the infection from cats; however, this drug has a narrow safety margin and clinical resistance is increasingly recognized. The more we learn about trichomonosis in cats, the more complicated and controversial the infection has become, ranging from what we should call the organism to whether we should even bother trying to treat it. Global importance: Feline trichomonosis is recognized to occur worldwide and is regarded as one of the most common infectious causes of colitis in the domestic cat. The infection is widespread in catteries and shelters; and, while remission of diarrhea may occur over time, persistence of the infection is common. Evidence base: This review provides a comprehensive examination of what is currently known about feline trichomonosis and pinpoints areas, based on the authors' opinion, where further research is needed.}, number={3}, journal={JOURNAL OF FELINE MEDICINE AND SURGERY}, author={Gookin, Jody L. and Hanrahan, Katherine and Levy, Michael G.}, year={2017}, month={Mar}, pages={261–274} }
 @article{schreeg_marr_tarigo_cohn_bird_scholl_levy_wiegmann_birkenheuer_2016, title={Mitochondrial Genome Sequences and Structures Aid in the Resolution of Piroplasmida phylogeny}, volume={11}, ISSN={["1932-6203"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-84994744879&partnerID=MN8TOARS}, DOI={10.1371/journal.pone.0165702}, abstractNote={The taxonomy of the order Piroplasmida, which includes a number of clinically and economically relevant organisms, is a hotly debated topic amongst parasitologists. Three genera (Babesia, Theileria, and Cytauxzoon) are recognized based on parasite life cycle characteristics, but molecular phylogenetic analyses of 18S sequences have suggested the presence of five or more distinct Piroplasmida lineages. Despite these important advancements, a few studies have been unable to define the taxonomic relationships of some organisms (e.g. C. felis and T. equi) with respect to other Piroplasmida. Additional evidence from mitochondrial genome sequences and synteny should aid in the inference of Piroplasmida phylogeny and resolution of taxonomic uncertainties. In this study, we have amplified, sequenced, and annotated seven previously uncharacterized mitochondrial genomes (Babesia canis, Babesia vogeli, Babesia rossi, Babesia sp. Coco, Babesia conradae, Babesia microti-like sp., and Cytauxzoon felis) and identified additional ribosomal fragments in ten previously characterized mitochondrial genomes. Phylogenetic analysis of concatenated mitochondrial and 18S sequences as well as cox1 amino acid sequence identified five distinct Piroplasmida groups, each of which possesses a unique mitochondrial genome structure. Specifically, our results confirm the existence of four previously identified clades (B. microti group, Babesia sensu stricto, Theileria equi, and a Babesia sensu latu group that includes B. conradae) while supporting the integration of Theileria and Cytauxzoon species into a single fifth taxon. Although known biological characteristics of Piroplasmida corroborate the proposed phylogeny, more investigation into parasite life cycles is warranted to further understand the evolution of the Piroplasmida. Our results provide an evolutionary framework for comparative biology of these important animal and human pathogens and help focus renewed efforts toward understanding the phylogenetic relationships within the group.}, number={11}, journal={PLOS ONE}, author={Schreeg, Megan E. and Marr, Henry S. and Tarigo, Jaime L. and Cohn, Leah A. and Bird, David M. and Scholl, Elizabeth H. and Levy, Michael G. and Wiegmann, Brian M. and Birkenheuer, Adam J.}, year={2016}, month={Nov} }
 @article{schreeg_marr_griffith_tarigo_bird_reichard_cohn_levy_birkenheuer_2016, title={PCR amplification of a multi-copy mitochondrial gene (cox3) improves detection of Cytauxzoon felis infection as compared to a ribosomal gene (18S)}, volume={225}, ISSN={["1873-2550"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-84975504702&partnerID=MN8TOARS}, DOI={10.1016/j.vetpar.2016.06.013}, abstractNote={Cytauxzoon felis is a tick-transmitted protozoan parasite that infects felids. Clinical disease caused by acute C. felis infection rapidly progresses in domestic cats, leading to high morbidity and mortality. Accurately diagnosing cytauxzoonosis as soon as possible during acute infection would allow for earlier initiation of antiprotozoal therapy which could lead to higher survival rates. Molecular detection of parasite rRNA genes (18S) by PCR has previously been shown to be a sensitive method of diagnosing C. felis infections. Based on evidence from related apicomplexan species, we hypothesized that C. felis mitochondrial genes would exist at higher copy numbers than 18S and would be a more sensitive diagnostic target. In this study we have designed a PCR assay targeting the C. felis mitochondrial gene cytochrome c oxidase subunit III (cox3). Herein we demonstrate that (1) the cox3 PCR can detect as low as 1 copy of DNA target and can detect C. felis in samples with known mitochondrial sequence heterogeneity, (2) cox3 copy number is increased relative to 18S in blood and tissue samples from acutely infected cats, and (3) the cox3 PCR is more sensitive than 18S PCR for detection of C. felis during early infections.}, journal={VETERINARY PARASITOLOGY}, author={Schreeg, Megan E. and Marr, Henry S. and Griffith, Emily H. and Tarigo, Jaime L. and Bird, David M. and Reichard, Mason V. and Cohn, Leah A. and Levy, Michael G. and Birkenheuer, Adam J.}, year={2016}, month={Jul}, pages={123–130} }
 @article{schreeg_marr_tarigo_cohn_levy_birkenheuer_2015, title={Rapid High-Resolution Melt Analysis of Cytauxzoon felis Cytochrome b To Aid in the Prognosis of Cytauxzoonosis}, volume={53}, ISSN={["1098-660X"]}, DOI={10.1128/jcm.00635-15}, abstractNote={Cytauxzoon felis is a virulent, tick-transmitted, protozoan parasite that infects felines. Cytauxzoonosis was previously thought to be uniformly fatal in domestic cats. Treatment combining atovaquone and azithromycin (A&A) has been associated with survival rates of over 60%. Atovaquone, a ubiquinone analogue, targets C. felis cytochrome b (cytb), of which 30 unique genotypes have been identified. The C. felis cytb genotype cytb1 is associated with increased survival rates in cats treated with A&A. The purpose of this study was to design a PCR panel that could distinguish C. felis cytb1 from other cytochrome b genotypes. Primer pairs were designed to span five different nucleotide positions at which single-nucleotide polymorphisms in the C. felis cytb gene had been identified. Through the use of high-resolution melt analysis, this panel was predicted to distinguish cytb1 from other cytb genotypes. Assays were validated using samples from 69 cats with cytauxzoonosis for which the C. felis cytb genotypes had been characterized previously. The PCR panel identified C. felis cytb1 with 100% sensitivity and 98.2% specificity. High-resolution melt analysis can rapidly provide prognostic information for clients considering A&A treatment in cats with cytauxzoonosis.}, number={8}, journal={JOURNAL OF CLINICAL MICROBIOLOGY}, author={Schreeg, Megan E. and Marr, Henry S. and Tarigo, Jaime L. and Cohn, Leah A. and Levy, Michael G. and Birkenheuer, Adam J.}, year={2015}, month={Aug}, pages={2517–2524} }
 @article{levy_powers_gore_marr_2015, title={Spironucleus meleagridis, an enteric diplomonad protozoan of cockatiels (Nymphicus hollandicus): Preliminary molecular characterization and association with clinical disease}, volume={208}, ISSN={["1873-2550"]}, DOI={10.1016/j.vetpar.2014.12.028}, abstractNote={A flagellated enteric diplomonad protozoan consistent with Spironucleus meleagridis (formerly Hexamita meleagridis) associated with gastrointestinal disease and mortality in psittacine birds including cockatiels (Nymphicus hollandicus) has been sporadically described in the literature. However, molecular characterization of psittacine protozoal isolates had not yet been performed. The 16S rRNA gene from a protozoan persistently shed in the feces in a small group of cockatiels demonstrated a 98% molecular identity with S. meleagridis isolated from turkeys. Based on these sequence data, a diagnostic PCR assay was developed to detect the presence of S. meleagridis. Nineteen privately owned pet cockatiels from unrelated households were clinically evaluated. All birds microscopically positive for this organism were PCR positive, with several additional birds microscopically negative but PCR positive. Many of the birds identified as positive for S. meleagridis by fecal PCR had signs of gastrointestinal disease such as diarrhea, soft feces, and melena, whereas none of the birds that tested negative had gastrointestinal signs. Examination of feces from two unrelated cockatiel breeding facilities revealed 70% and 86% PCR positive rates. Prevalence of infection and incidence of clinical disease, including factors that lead to clinical manifestation such as viral, bacterial, or mycotic coinfections, are not yet known and warrant further study, but spironucleosis is likely an under-recognized disease in cockatiels.}, number={3-4}, journal={VETERINARY PARASITOLOGY}, author={Levy, M. G. and Powers, L. V. and Gore, K. C. and Marr, H. S.}, year={2015}, month={Mar}, pages={169–173} }
 @article{ghirardi_levy_lopez_corbalan_steciow_perotti_2014, title={Endangered amphibians infected with the chytrid fungus Batrachochytrium dendrobatidis in austral temperate wetlands from Argentina}, volume={24}, number={2}, journal={Herpetological Journal}, author={Ghirardi, R. and Levy, M. G. and Lopez, J. A. and Corbalan, V. and Steciow, M. M. and Perotti, M. G.}, year={2014}, pages={129–133} }
 @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{li_d’annibale-tolhurst_adler_fang_yin_birkenheuer_levy_jones_sung_hawkins_et al._2013, title={A Myristoylated Alanine-Rich C Kinase Substrate–Related Peptide Suppresses Cytokine mRNA and Protein Expression in LPS-Activated Canine Neutrophils}, volume={48}, ISSN={1044-1549 1535-4989}, url={http://dx.doi.org/10.1165/rcmb.2012-0278OC}, DOI={10.1165/rcmb.2012-0278oc}, abstractNote={Section:ChooseTop of pageAbstract <<Materials and MethodsResultsDiscussionReferencesCITING ARTICLES}, number={3}, journal={American Journal of Respiratory Cell and Molecular Biology}, publisher={American Thoracic Society}, author={Li, Jingjing and D’Annibale-Tolhurst, Melissa A. and Adler, Kenneth B. and Fang, Shijing and Yin, Qui and Birkenheuer, Adam J. and Levy, Michael G. and Jones, Samuel L. and Sung, Eui Jae and Hawkins, Eleanor C. and et al.}, year={2013}, month={Mar}, pages={314–321} }
 @article{tarigo_scholl_bird_brown_cohn_dean_levy_doolan_trieu_nordone_et al._2013, title={A Novel Candidate Vaccine for Cytauxzoonosis Inferred from Comparative Apicomplexan Genomics}, volume={8}, ISSN={["1932-6203"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-84882652524&partnerID=MN8TOARS}, DOI={10.1371/journal.pone.0071233}, abstractNote={Cytauxzoonosis is an emerging infectious disease of domestic cats (Felis catus) caused by the apicomplexan protozoan parasite Cytauxzoon felis. The growing epidemic, with its high morbidity and mortality points to the need for a protective vaccine against cytauxzoonosis. Unfortunately, the causative agent has yet to be cultured continuously in vitro, rendering traditional vaccine development approaches beyond reach. Here we report the use of comparative genomics to computationally and experimentally interpret the C. felis genome to identify a novel candidate vaccine antigen for cytauxzoonosis. As a starting point we sequenced, assembled, and annotated the C. felis genome and the proteins it encodes. Whole genome alignment revealed considerable conserved synteny with other apicomplexans. In particular, alignments with the bovine parasite Theileria parva revealed that a C. felis gene, cf76, is syntenic to p67 (the leading vaccine candidate for bovine theileriosis), despite a lack of significant sequence similarity. Recombinant subdomains of cf76 were challenged with survivor-cat antiserum and found to be highly seroreactive. Comparison of eleven geographically diverse samples from the south-central and southeastern USA demonstrated 91-100% amino acid sequence identity across cf76, including a high level of conservation in an immunogenic 226 amino acid (24 kDa) carboxyl terminal domain. Using in situ hybridization, transcription of cf76 was documented in the schizogenous stage of parasite replication, the life stage that is believed to be the most important for development of a protective immune response. Collectively, these data point to identification of the first potential vaccine candidate antigen for cytauxzoonosis. Further, our bioinformatic approach emphasizes the use of comparative genomics as an accelerated path to developing vaccines against experimentally intractable pathogens.}, number={8}, journal={PLOS ONE}, author={Tarigo, Jaime L. and Scholl, Elizabeth H. and Bird, David McK and Brown, Corrie C. and Cohn, Leah A. and Dean, Gregg A. and Levy, Michael G. and Doolan, Denise L. and Trieu, Angela and Nordone, Shila K. and et al.}, year={2013}, month={Aug} }
 @article{hernandez_galbreath_riddle_moore_palamar_levy_deperno_correa_yabsley_2013, title={Baylisascaris procyonis in raccoons (Procyon lotor) from North Carolina and current status of the parasite in the USA}, volume={112}, ISSN={0932-0113 1432-1955}, url={http://dx.doi.org/10.1007/s00436-012-3186-1}, DOI={10.1007/s00436-012-3186-1}, number={2}, journal={Parasitology Research}, publisher={Springer Science and Business Media LLC}, author={Hernandez, Sonia M. and Galbreath, Brianna and Riddle, Dennis F. and Moore, Andrew P. and Palamar, Maria B. and Levy, Michael G. and DePerno, Christopher S. and Correa, Maria T. and Yabsley, Michael J.}, year={2013}, month={Nov}, pages={693–698} }
 @article{schreeg_marr_tarigo_cohn_levy_birkenheuer_2013, title={Pharmacogenomics of Cytauxzoon felis Cytochrome b: Implications for Atovaquone and Azithromycin Therapy in Domestic Cats with Cytauxzoonosis}, volume={51}, ISSN={["0095-1137"]}, DOI={10.1128/jcm.01407-13}, abstractNote={Cytauxzoon felis, an emerging virulent protozoan parasite that infects domestic cats, is treated with atovaquone and azithromycin (A&A). Atovaquone targets parasite cytochrome b. We characterized the C. felis cytochrome b gene (cytb) in cats with cytauxzoonosis and found a cytb genotype that was associated with survival in A&A-treated cats.}, number={9}, journal={JOURNAL OF CLINICAL MICROBIOLOGY}, author={Schreeg, Megan E. and Marr, Henry S. and Tarigo, Jaime and Cohn, Leah A. and Levy, Michael G. and Birkenheuer, Adam J.}, year={2013}, month={Sep}, pages={3066–3069} }
 @article{billeter_kasten_killmaster_breitschwerdt_levin_levy_kosoy_chomel_2012, title={Experimental infection by capillary tube feeding of Rhipicephalus sanguineus with Bartonella vinsonii subspecies berkhoffii}, volume={35}, ISSN={0147-9571}, url={http://dx.doi.org/10.1016/j.cimid.2011.09.004}, DOI={10.1016/j.cimid.2011.09.004}, abstractNote={It has been speculated that ticks may serve as vectors of Bartonella species. Circumstantial, clinical, epidemiological and serological evidence suggest that B. vinsonii subspecies berkhoffii (B. v. berkhoffii) might be transmitted by Rhipicephalus sanguineus. The purpose of the present study was to determine whether adult R. sanguineus ticks can be infected with a B. v. berkhoffii genotype II isolate via capillary tube feeding and whether the infection can then be transmitted from adult females to their eggs via trans-ovarial transmission. Furthermore, tick fecal material was also collected and screened as a possible source of infectious inoculum for canine infections. B. v. berkhoffii DNA was detected in 50% (7 of 14) of females that did not oviposit and in 14.3% (2 of 14) of female ticks that laid eggs, but not detected in egg clutches (100 eggs/female). DNA was also detected in tick feces collected on days 2 through 6 post-capillary tube feeding, however, dogs (n=3) did not become bacteremic or seroconvert when inoculated with tick fecal material. Therefore, trans-ovarial transmission of B. v. berkhoffii by R. sanguineus is unlikely, but further studies are needed to determine if tick fecal material can serve as a source of infection to canines.}, number={1}, journal={Comparative Immunology, Microbiology and Infectious Diseases}, publisher={Elsevier BV}, author={Billeter, Sarah A. and Kasten, Rick W. and Killmaster, Lindsay F. and Breitschwerdt, Edward B. and Levin, Michael L. and Levy, Michael G. and Kosoy, Michael Y. and Chomel, Bruno B.}, year={2012}, month={Jan}, pages={9–15} }
 @article{billeter_breitschwerdt_levy_2012, title={Invasion of canine erythrocytes by Bartonella vinsonii subsp. berkhoffii}, volume={156}, ISSN={0378-1135}, url={http://dx.doi.org/10.1016/j.vetmic.2011.10.020}, DOI={10.1016/j.vetmic.2011.10.020}, abstractNote={Bartonella vinsonii subsp. berkhoffii is a recognized cause of endocarditis in dogs and human patients and has been associated with cardiac arrhythmias, myocarditis, granulomatous lymphadenitis, polyarthritis, and granulomatous rhinitis in dogs. Little is known regarding the mode of transmission or cellular localization of this bacteria following infection of a canine host. The aim of the current study was to determine whether erythrocytes may serve as a site of infection by B. vinsonii subsp. berkhoffii. In the study, we successfully demonstrate the invasion of canine erythrocytes by a B. vinsonii subsp. berkhoffii genotype III strain using an in vitro model system. Dog erythrocytes were incubated with B. vinsonii subsp. berkhoffii after which tubes were treated with gentamicin at 12, 24, and 48 h post-inoculation. After gentamicin elimination of extracellular bacteria, there was a gradual increase in intra-erythrocytic bacteria, as assessed by colony forming units per ml, at each collection time point. The largest recovery of intracellular bacteria occurred at 48 h post-infection. These results suggest that canine erythrocytes may serve in the maintenance of bacteremia due to B. vinsonii subsp. berkhoffii within an infected host.}, number={1-2}, journal={Veterinary Microbiology}, publisher={Elsevier BV}, author={Billeter, Sarah A. and Breitschwerdt, Edward B. and Levy, Michael G.}, year={2012}, month={Apr}, pages={213–216} }
 @article{greenspan_calhoun_longcore_levy_2012, title={TRANSMISSION OF BATRACHOCHYTRIUM DENDROBATIDIS TO WOOD FROGS (LITHOBATES SYLVATICUS) VIA A BULLFROG (L. CATESBEIANUS) VECTOR}, volume={48}, ISSN={["1943-3700"]}, DOI={10.7589/0090-3558-48.3.575}, abstractNote={Chytridiomycosis, an emerging infectious disease caused by the chytrid fungus Batrachochytrium dendrobatidis, threatens anuran populations worldwide. Effects of B. dendrobatidis on frog species are variable. Some species typically develop nonlethal infections and may function as carriers; others typically develop lethal infections that can lead to population declines. Nonlethal infections in the bullfrog (Lithobates catesbeianus) are well-documented. In contrast, recently metamorphosed wood frogs (L. sylvaticus) can die from chytridiomycosis. We conducted an ex-situ experiment between May and July 2010 to determine whether B. dendrobatidis-infected bullfrogs could transmit the fungus to wood frog tadpoles when the two species shared a body of water. We tested for B. dendrobatidis infections with quantitative polymerase chain reactions (qPCR) in a subsample of the wood frog tadpoles and in all metamorphosed wood frogs and compared risk of death of froglets exposed and unexposed to infected bullfrogs. We detected B. dendrobatidis sporadically in subsampled treatment tadpoles (nine of 90, 10%) and frequently in treatment froglets (112 of 113, 99.1%). Pooled risk of froglet death was higher (P<0.001) in treatment enclosures than in control enclosures. Our results indicate that, at the low infection loads bullfrogs tend to carry, swabbing for PCR analyses may underestimate prevalence of B. dendrobatidis in this species. We highlight bullfrog disease screening as a management challenge, especially in light of exotic bullfrog colonies on multiple continents and large-scale global trade in this species. We document the importance of quantifying lethal and sublethal effects of bullfrog vectors on B. dendrobatidis-susceptible species.}, number={3}, journal={JOURNAL OF WILDLIFE DISEASES}, author={Greenspan, Sasha E. and Calhoun, Aram J. K. and Longcore, Joyce E. and Levy, Michael G.}, year={2012}, month={Jul}, pages={575–582} }
 @article{li_birkenheuer_marr_levy_yoder_nordone_2011, title={Expression and function of triggering receptor expressed on myeloid cells-1 (TREM-1) on canine neutrophils}, volume={35}, ISSN={0145-305X}, url={http://dx.doi.org/10.1016/j.dci.2011.03.021}, DOI={10.1016/j.dci.2011.03.021}, abstractNote={The dog is both a valued veterinary species and a widely used translational model for sepsis research. However, relatively little work has been performed evaluating potential biomarkers present during canine infection. Triggering receptor expressed on myeloid cells-1 (TREM-1) has shown promise as a biomarker for infection and pneumonia in humans. Here we describe, for the first time, the expression and function of the canine orthologue of TREM-1. Expression of TREM-1 on canine neutrophils is significantly up-regulated by stimulation with microbial agonists of TLR2/6, TLR1/2, and TLR4/MD2. Kinetics of TREM-1 protein up-regulation are rapid, with significant increases observed within 2 hr of neutrophil activation. Functionally, canine TREM-1 synergistically enhances LPS-induced production of IL-8, TNF-α and a canine orthologue of CXCL1. Collectively, these data suggest that TREM-1 expression in dogs, as it is in humans, is an amplifier of pro-inflammatory responses to microbial products. These results have direct application to veterinary diagnostics as well as the potential to enhance the utility of canine disease models in the assessment of potential therapeutics in the treatment of human sepsis.}, number={8}, journal={Developmental & Comparative Immunology}, publisher={Elsevier BV}, author={Li, Jingjing and Birkenheuer, Adam J. and Marr, Henry S. and Levy, Michael G. and Yoder, Jeffrey A. and Nordone, Shila K.}, year={2011}, month={Aug}, pages={872–880} }
 @article{chinnadurai_cooper_dombrowski_poore_levy_2009, title={EXPERIMENTAL INFECTION OF NATIVE NORTH CAROLINA SALAMANDERS WITH BATRACHOCHYTRIUM DENDROBATIDIS}, volume={45}, ISSN={["0090-3558"]}, DOI={10.7589/0090-3558-45.3.631}, abstractNote={Chytridiomycosis is an often fatal fungal disease of amphibians caused by Batrachochytrium dendrobatidis. This disease has been implicated in the worldwide decline of many anuran species, but studies of chytridiomycosis in wild salamanders are limited. Between August 2006 and December 2006, we tested wild amphibians in North Carolina, USA (n=212) by polymerase chain reaction (PCR). We identified three PCR-positive animals: one Rana clamitans and two Plethodontid salamanders. We experimentally infected two species of native North Carolina Plethodontid salamanders, the slimy salamander (Plethodon glutinosus) and the Blue Ridge Mountain dusky salamander (Desmognathus orestes) with 1,000,000 zoospores of B. dendrobatidis per animal. Susceptibility was species dependent; all slimy salamanders developed clinical signs of chytridiomycosis, and one died, whereas dusky salamanders remained unaffected. In a second experiment, we challenged naïve slimy salamanders with either 10,000 or 100,000 motile zoospores per animal. Clinical signs consistent with chytridiomycosis were not observed at either dose or in uninfected controls during the 45 days of this experiment. All animals inoculated with B. dendrobatidis in both experiments, regardless of dose, tested positive by PCR. Our study indicates that slimy salamanders are more susceptible to clinical chytridiomycosis than dusky salamanders, and in a laboratory setting, a dose greater than 100,000 zoospores per animal is required to induce clinical disease. This study also indicates that PCR is a very sensitive tool for detecting B. dendrobatidis infection, even in animals that are clinically unaffected, thus positive results should be interpreted with caution.}, number={3}, journal={JOURNAL OF WILDLIFE DISEASES}, author={Chinnadurai, Sathya K. and Cooper, David and Dombrowski, Daniel S. and Poore, Matthew F. and Levy, Michael G.}, year={2009}, month={Jul}, pages={631–636} }
 @article{billeter_diniz_battisti_munderloh_breitschwerdt_levy_2009, title={Infection and replication of Bartonella species within a tick cell line}, volume={49}, ISSN={0168-8162 1572-9702}, url={http://dx.doi.org/10.1007/s10493-009-9255-1}, DOI={10.1007/s10493-009-9255-1}, abstractNote={Bartonella species are fastidious, gram negative bacteria, some of which are transmitted by arthropod vectors, including fleas, sandflies, and lice. There is very little information regarding the interaction and/or transmission capabilities of Bartonella species by ticks. In the present study, we demonstrate successful infection of the Amblyomma americanum cell line, AAE12, by seven Bartonella isolates and three Candidatus Bartonella species by electron or light microscopy. With the exception of Bartonella bovis, infection with all other examined Bartonella species induced cytopathic effects characterized by heavy cellular vacuolization and eventually cell lysis. Furthermore, using quantitative real time PCR (qPCR), we demonstrated significant amplification of two B. henselae genotype I isolates in the A. americanum cell line over a 5 days period. Ultimately, tick-cell derived Bartonella antigens may prove useful for the development of more sensitive diagnostic reagents and may assist in the development of an effective vaccine to prevent the further spread of disease caused by these organisms.}, number={3}, journal={Experimental and Applied Acarology}, publisher={Springer Science and Business Media LLC}, author={Billeter, Sarah A. and Diniz, Pedro Paulo V. P. and Battisti, James M. and Munderloh, Ulrike G. and Breitschwerdt, Edward B. and Levy, Michael G.}, year={2009}, month={Feb}, pages={193–208} }
 @article{billeter_miller_breitschwerdt_levy_2008, title={Detection of two Bartonella tamiae-like sequences in Amblyomma americanum (Acari : Ixodidae) using 16S-23S intergenic spacer region-specific primers}, volume={45}, ISSN={["0022-2585"]}, DOI={10.1603/0022-2585(2008)45[176:DOTBTS]2.0.CO;2}, abstractNote={Four hundred and sixty-six questing Amblyomma americanum (L.) (Acari: Ixodidae) from Carolina County, VA, and 98 questing A. americanum from Chatham County, NC, were screened by polymerase chain reaction (PCR) for the Bartonella 16S-23S intergenic spacer region. Two amplicons, approximately 270-280 bp, were detected in two ticks from Virginia. Based upon PCR and sequencing, an adult male and adult female tick harbored DNA sequences closely related to Bartonella tamiae (DQ395180). Bartonella DNA was not detected in A. americanum from North Carolina. Potential transmission of Bartonella spp. by A. americanum should be the focus of future experimental studies.}, number={1}, journal={JOURNAL OF MEDICAL ENTOMOLOGY}, author={Billeter, Sarah A. and Miller, Melissa K. and Breitschwerdt, Edward B. and Levy, Michael G.}, year={2008}, month={Jan}, pages={176–179} }
 @article{stauffer_birkenheuer_levy_marr_gookin_2008, title={Evaluation of four DNA extraction methods for the detection of Tritrichomonas foetus in feline stool specimens by polymerase chain reaction}, volume={20}, ISSN={["1943-4936"]}, DOI={10.1177/104063870802000518}, abstractNote={Feces are increasingly valued as practical samples for molecular diagnosis of infectious disease. However, extraction of polymerase chain reaction (PCR) quality DNA from fecal samples can be challenging because of coextraction of PCR inhibitors. Because the type and quantity of PCR inhibitors is influenced by diet, endogenous flora, and concurrent disease, it is unlikely that extraction method performance with human feces can be directly extrapolated to that of domestic cats. In the present study, 4 commercially available DNA extraction methods were examined for their influence on the sensitivity of PCR for the detection of Tritrichomonas foetus in feline stool. DNA was extracted from serially diluted feline-origin T. foetus trophozoites in the absence or presence of feline feces. The ZR Fecal DNA kit was identified as affording the greatest analytical sensitivity and reproducibility and was able to detect ≥10 T. foetus organisms per 100 mg feces in 100% of PCR reactions. Further, the identified extraction method could be completed in the shortest time of all kits tested.}, number={5}, journal={JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION}, author={Stauffer, Stephen H. and Birkenheuer, Adam J. and Levy, Michael G. and Marr, Henry and Gookin, Jody L.}, year={2008}, month={Sep}, pages={639–641} }
 @article{levy_crawford_lappin_dubovi_levy_alleman_tucker_clifford_2008, title={Infectious diseases of dogs and cats on Isabela Island, Galapagos}, volume={22}, ISSN={["1939-1676"]}, DOI={10.1111/j.1939-1676.2007.0034.x}, abstractNote={Background: Vaccination and importation of dogs and cats are prohibited in the Galapagos, resulting in a uniquely isolated population. The purpose of this study was to determine the prevalence of infectious diseases of dogs and cats that impact their health, could spill over to native wildlife, or sentinel diseases of concern to humans. Hypothesis: The isolation of dogs and cats in the Galapagos protects them from diseases common in mainland populations. Animals: Ninety-five dogs and 52 cats presented during a neutering campaign. Methods: A prospective cross-sectional study was performed. Blood was collected for serological and DNA evaluation of a panel of infectious diseases. Results: Antibodies against parvovirus (100%), parainfluenza virus (100%), adenovirus 1/2 (66–67%), and distemper virus (22%) were present in dogs. Dirofilaria immitis was also common in dogs (34%), with lower prevalences of Wolbachia pipiens (22%), Bartonella sp. (13%), Ehrlichia/Anaplasma spp. (1%), and Mycoplasma haemocanis (1%) observed. Antibodies against panleukopenia virus (67%), Toxoplasma gondii (63%), calicivirus (44%), and herpesvirus 1 (10%) were detected in cats. Feline leukemia virus antigen, feline immunodeficiency virus antibody, or coronavirus antibodies were not detected. Bartonella sp. (44%) infections were common in cats, but only one was infected with M. haemofelis. Conclusions and Clinical Importance: Despite their relative seclusion from the rest of the world, cats and dogs of Isabela were exposed to many pathogens found in mainland South America. Parasite prophylaxis, neutering, and strict enforcement of animal movement restrictions would control a majority of the diseases. In the absence of vaccination, a reservoir of susceptible animals remains vulnerable to new disease introductions.}, number={1}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Levy, J. K. and Crawford, P. C. and Lappin, M. R. and Dubovi, E. J. and Levy, M. G. and Alleman, R. and Tucker, S. J. and Clifford, E. L.}, year={2008}, pages={60–65} }
 @article{billeter_levy_chomel_breitschwerdt_2008, title={Vector transmission of Bartonella species with emphasis on the potential for tick transmission}, volume={22}, ISSN={0269-283X 1365-2915}, url={http://dx.doi.org/10.1111/j.1365-2915.2008.00713.x}, DOI={10.1111/j.1365-2915.2008.00713.x}, abstractNote={Abstract Bartonella species are gram-negative bacteria that infect erythrocytes, endothelial cells and macrophages, often leading to persistent blood-borne infections. Because of the ability of various Bartonella species to reside within erythrocytes of a diverse number of animal hosts, there is substantial opportunity for the potential uptake of these blood-borne bacteria by a variety of arthropod vectors that feed on animals and people. Five Bartonella species are transmitted by lice, fleas or sandflies. However, Bartonella DNA has been detected or Bartonella spp. have been cultured from numerous other arthropods. This review discusses Bartonella transmission by sandflies, lice and fleas, the potential for transmission by other vectors, and data supporting transmission by ticks. Polymerase chain reaction (PCR) or culture methods have been used to detect Bartonella in ticks, either questing or host-attached, throughout the world. Case studies and serological or molecular surveys involving humans, cats and canines provide indirect evidence supporting transmission of Bartonella species by ticks. Of potential clinical relevance, many studies have proposed co-transmission of Bartonella with other known tick-borne pathogens. Currently, critically important experimental transmission studies have not been performed for Bartonella transmission by many potential arthropod vectors, including ticks.}, number={1}, journal={Medical and Veterinary Entomology}, publisher={Wiley}, author={Billeter, S. A. and Levy, M. G. and Chomel, B. B. and Breitschwerdt, E. B.}, year={2008}, month={Mar}, pages={1–15} }
 @article{levy_poore_colorni_noga_vandersea_litaker_2007, title={A highly specific PCR assay for detecting the fish ectoparasite Amyloodinium ocellatum}, volume={73}, ISSN={["1616-1580"]}, DOI={10.3354/dao073219}, abstractNote={Amyloodiniosis, caused by the dinoflagellate ectoparasite Amyloodinium ocellatum, is one of the most serious diseases affecting marine fish in warm and temperate waters. Current diagnostic methods rely entirely on the microscopic identification of parasites on the skin or gills of infested fish. However, subclinical infestations usually go undetected, while no method of detecting the free-swimming, infective (dinospore) stage has been devised. Targeting the parasite's ribosomal DNA region, we have developed a sensitive and specific PCR assay that can detect as little as a single cell from any of the 3 stages of the parasite's life cycle (trophont, tomont, dinospore). This assay performs equally well in a simple artificial seawater medium and in natural seawater containing a plankton community assemblage. The assay is also not inhibited by gill tissue. Sequence analysis of the internal transcribed spacer region of 5 A. ocellatum isolates, obtained from fish in the Red Sea (Israel), eastern Mediterranean Sea (Israel), Adriatic Sea (Italy), Gulf of Mexico (Florida), and from an unknown origin, revealed insignificant variation, indicating that all isolates were the same species. However, 3 of these isolates propagated in cell culture varied in behavior and morphology, and these differences were consistent during at least 2 yr in culture. Thus, our findings do not eliminate the possibility that different strains are in fact 'subspecies' or lower taxa, which may also differ in pathogenic and immunogenic characteristics, environmental tolerance, and other features.}, number={3}, journal={DISEASES OF AQUATIC ORGANISMS}, author={Levy, Michael G. and Poore, Matthew F. and Colorni, Angelo and Noga, Edward J. and Vandersea, Mark W. and Litaker, R. Wayne}, year={2007}, month={Jan}, pages={219–226} }
 @article{billeter_blanton_little_levy_breitschwerdt_2007, title={Detection of “Rickettsia amblyommii” in Association with a Tick Bite Rash}, volume={7}, ISSN={1530-3667 1557-7759}, url={http://dx.doi.org/10.1089/vbz.2007.0121}, DOI={10.1089/vbz.2007.0121}, abstractNote={In the summer of 2006, an Amblyomma americanum tick was removed from a woman in central North Carolina, who subsequently developed a rash at the site of tick attachment. When examined by polymerase chain reaction (PCR) for Borrelia, Anaplasma, Ehrlichia, Babesia, Rickettsia, and Bartonella DNA, only the Rickettsia primers generated an amplicon, which was identified as “R. amblyommii” by sequencing. To our knowledge, this is the first case in which R. amblyommii was temporally associated with a rash.}, number={4}, journal={Vector-Borne and Zoonotic Diseases}, publisher={Mary Ann Liebert Inc}, author={Billeter, Sarah A. and Blanton, Hunter L. and Little, Susan E. and Levy, Michael G. and Breitschwerdt, Edward B.}, year={2007}, month={Dec}, pages={607–610} }
 @article{gookin_stauffer_coccaro_poore_levy_papich_2007, title={Efficacy of tinidazole for treatment of cats experimentally infected with Tritrichomonas foetus}, volume={68}, ISSN={["1943-5681"]}, DOI={10.2460/ajvr.68.10.1085}, abstractNote={Abstract Objective —To determine the efficacy of tinidazole for treatment of cats with experimentally induced Tritrichomonas foetus infection. Animals —8 specific-pathogen-free kittens. Procedures —Tinidazole was tested for activity against a feline isolate of T foetus in vitro. Kittens were infected orogastrically with the same isolate and treated or not with tinidazole (30 mg/kg, PO, q 24 h for 14 days). Amoxicillin was administered 28 weeks after completion of tinidazole administration to induce diarrhea. Feces were repeatedly tested for T foetus by use of PCR assay and microbial culture for 33 weeks. Results —Tinidazole killed T foetus at concentrations ≥ 10 μg/mL in vitro. In experimentally induced infection, tinidazole administered at 30 mg/kg decreased T foetus below the limit of molecular detection in 2 of 4 cats. Recrudescent shedding of T foetus , as elicited by amoxicillin-induced diarrhea, was diminished in cats that received prior treatment with tinidazole. Conclusions and Clinical Relevance —Although tinidazole decreased the detection of T foetus and treated cats were resistant to later efforts to incite the infection, inability of tinidazole to eradicate infection in many cats poses a serious impediment to the drug’s effectiveness in practice.}, number={10}, journal={AMERICAN JOURNAL OF VETERINARY RESEARCH}, author={Gookin, Jody L. and Stauffer, Stephen H. and Coccaro, Maria R. and Poore, Matthew F. and Levy, Michael G. and Papich, Mark G.}, year={2007}, month={Oct}, pages={1085–1088} }
 @article{gookin_stauffer_levy_2007, title={Identification of Pentatrichomonas hominis in feline fecal samples by polymerase chain reaction assay}, volume={145}, ISSN={["1873-2550"]}, DOI={10.1016/j.vetpar.2006.10.020}, abstractNote={Pentatrichomonas hominis is considered to be a commensal protozoan of the vertebrate digestive tract. On the basis of light microscopic examination of feces, some investigators presumptively identified P. hominis as a causative agent of feline diarrhea. However, molecular identification of P. hominis infection in the cat has not been reported. Another trichomonad, Tritrichomonas foetus, is recognized as an intestinal pathogen in cats and often presumptively diagnosed on the basis of the presence of trichomonads in diarrheic feces. It is of importance to determine if cats are natural hosts for P. hominis, as the presence of this organism could result in inaccurate assumption of T. foetus infection. In this study, we used a species-specific PCR assay to identify P. hominis 18S rRNA genes in fecal samples collected from a convenience population of cats in which a high prevalence of T. foetus infection had been previously identified (cat show) or suspected (submitted for T. foetus diagnostic testing). The prevalence of T. foetus infection in these samples was 31% and 28.6%, respectively. P. hominis infection was identified by PCR of DNA extracted from feces of five cats (1.9% and 2.1% of fecal samples, respectively). All cats in which P. hominis was identified were also infected with T. foetus. PCR identification of P. hominis infection in the cat should facilitate future studies to determine the pathogenicity of this species and enable differentiation of P. hominis from other known or as-yet unidentified species of trichomonads that may infect cats.}, number={1-2}, journal={VETERINARY PARASITOLOGY}, author={Gookin, Jody L. and Stauffer, Stephen H. and Levy, Michael G.}, year={2007}, month={Apr}, pages={11–15} }
 @article{gookin_stauffer_coccaro_marcotte_levy_2007, title={Optimization of a species-specific polymerase chain reaction assay for identification of Pentatrichomonas hominis in canine fecal specimens}, volume={68}, ISSN={["1943-5681"]}, DOI={10.2460/ajvr.68.7.783}, abstractNote={Abstract Objective —To determine the optimum reaction conditions and detection limits of PCR assay for identification of Pentatrichomonas hominis in DNA extracted from canine feces. Sample Population —DNA extracted from feces of 4 dogs with diarrhea from which trichomonads were observed, 81 dogs that had feces submitted to a diagnostic laboratory, and 19 dogs residing in a laboratory animal facility. Procedures —Optimum reaction conditions and absolute and practical detection limits of 2 P hominis 18S species-specific primer pairs were determined by use of an in vitro cultivated canine isolate of P hominis in the presence and absence of canine feces. The optimized PCR assay was applied to amplification of P hominis 18S rRNA genes from DNA extracted from the feces of dogs. Results —Under optimized conditions, a primer pair was identified as able to detect as few as 1 P hominis organism/180-mg fecal sample. The PCR assay identified P hominis in diarrheic feces of 4 dogs in which trichomonads were seen by light microscopy. The P hominis genes were not amplified from other fecal samples examined. Conclusions and Clinical Relevance —Molecular identification of P hominis in feces of 4 dogs with trichomonosis and diarrhea reported here validates the identity of this species in such infections. Sensitive and specific PCR amplification of P hominis 18S rRNA genes from DNA extracted from feces will directly facilitate studies examining pathogenicity of this trichomonad and enable differentiation of P hominis from other known or novel species of trichomonads that may infect the gastrointestinal tract of dogs.}, number={7}, journal={AMERICAN JOURNAL OF VETERINARY RESEARCH}, author={Gookin, Jody L. and Stauffer, Stephen H. and Coccaro, Maria R. and Marcotte, Miriam J. and Levy, Michael G.}, year={2007}, month={Jul}, pages={783–787} }
 @article{levy_litaker_goldstein_dykstra_vandersea_noga_2007, title={Piscinoodinium, a fish-ectoparasitic dinoflagellate, is a member of the class Dinophyceae, subclass Gymnodiniphycidae: Convergent evolution with Amyloodinium}, volume={93}, ISSN={["1937-2345"]}, DOI={10.1645/GE-3585.1}, abstractNote={All dinoflagellates that infest the skin and gills of fish have traditionally been placed within the class Blastodiniphyceae. Their relatedness was primarily based upon a similar mode of attachment to the host, i.e., attachment disc with holdfasts. Results of recent molecular genetic analyses have transferred these parasites, including Amyloodinium, to the class Dinophyceae, subclass Peridiniphycidae. In our study, a small subunit rDNA gene from a parasitic dinoflagellate that has features diagnostic for species in the genus Piscinoodinium, i.e., typical trophont with attachment disc having rhizocysts, infesting the skin of freshwater tropical fish, places this organism within the dinophycean subclass Gymnodiniphycidae. This suggests a close relationship of Piscinoodinium spp. to dinoflagellates that include symbionts, e.g., species of Symbiodinium, and free-living algae, e.g., Gymnodinium spp. These molecular and morphological data suggest that evolution of this mode of fish ectoparasitism occurred independently in 2 distantly related groups of dinoflagellates, and they further suggest that the taxonomic status of parasites grouped as members of Piscinoodinium requires major revision.}, number={5}, journal={JOURNAL OF PARASITOLOGY}, author={Levy, Michael G. and Litaker, R. Wayne and Goldstein, Robert J. and Dykstra, Michael J. and Vandersea, Mark W. and Noga, Edward J.}, year={2007}, month={Oct}, pages={1006–1015} }
 @article{mitchell_goodwin_levy_2006, title={Bolbophorus infections in cultured fathead minnow}, volume={18}, ISSN={["0899-7659"]}, DOI={10.1577/h05-004.1}, abstractNote={Bolbophorus trematodes in fish recently gained national attention because B. damnificus infections have caused major losses of commercially raised channel catfish Ictalurus punctatus. We report the presence of a mixed infection (B. damnificus and Bolbophorus sp.) in fathead minnow Pimephales promelas. Of the 10 trematodes identified by Bolbophorus spp.−specific polymerase chain reaction assays, 9 were Bolbophorus sp. and 1 was B. damnificus. This is the first report of a natural infection of B. damnificus in a nonictalurid host. In fathead minnow, the prodiplostomula examined histologically were surrounded by thin-walled cysts; rarely, the cysts were accompanied by an inflammatory reaction that consisted primarily of macrophages.}, number={1}, journal={JOURNAL OF AQUATIC ANIMAL HEALTH}, author={Mitchell, AJ and Goodwin, AE and Levy, MG}, year={2006}, month={Mar}, pages={55–57} }
 @article{birkenheuer_le_valenzisi_tucker_levy_breitschwerdt_2006, title={Cytauxzoon felisinfection in cats in the mid-Atlantic states: 34 cases (1998–2004)}, volume={228}, ISSN={0003-1488}, url={http://dx.doi.org/10.2460/javma.228.4.568}, DOI={10.2460/javma.228.4.568}, abstractNote={Abstract Objective —To describe the demographic and clinical characteristics of feline cytauxzoonosis in the midAtlantic states and compare the Cytauxzoon felis 18S rRNA gene sequences from affected cats with sequences reported from affected cats in other regions. Design —Retrospective case series. Animals —34 cats with C felis infection. Procedure —Medical records of cats in which C felis infection was diagnosed from May 1998 through June 2004 were reviewed; data collected included signalment, month of diagnosis, geographic location, clinicopathologic abnormalities, medical treatments, outcome, and necropsy findings when applicable. Cytauxzoon felis DNA was amplified, cloned, and sequenced from 4 of these cats and compared with previously reported C felis DNA sequences. Results —Of 34 C felis –infected cats, 28 resided in North Carolina, 3 resided in South Carolina, and 3 resided in Virginia; in 32 cats, a diagnosis of C felis infection was made in April through September. Pancytopenia and icterus were the most common clinicopathologic abnormalities. Thirty-two cats either died or were euthanatized, and 2 cats survived. At 5 veterinary hospitals, multiple cases were identified, and 4 multicat households had > 1 cat infected with C felis . The 18S rRNA gene sequences characterized in organisms obtained from 4 cats were nearly identical to C felis DNA sequences reported from other US regions. Conclusions and Clinical Relevance —Data indicate that veterinarians in the mid-Atlantic region of the United States should consider C felis infection in cats that become ill with fever, icterus, and pancytopenia or bicytopenia, especially in the spring and summer months.}, number={4}, journal={Journal of the American Veterinary Medical Association}, publisher={American Veterinary Medical Association (AVMA)}, author={Birkenheuer, Adam J. and Le, Jaime A. and Valenzisi, Amy M. and Tucker, Melissa D. and Levy, Michael G. and Breitschwerdt, Edward B.}, year={2006}, month={Feb}, pages={568–571} }
 @article{birkenheuer_marr_alleman_levy_breitschwerdt_2006, title={Development and evaluation of a PCR assay for the detection of Cytauxzoon felis DNA in feline blood samples}, volume={137}, ISSN={["1873-2550"]}, DOI={10.1016/j.vetpar.2005.12.007}, abstractNote={Cytauxzoonosis is an emerging tick borne infectious disease of domestic cats in the United States, caused by the organism Cytauxzoon felis (C. felis). In naturally infected domestic cats the disease is almost always fatal. Currently there are no commercially available molecular or serologic tests to facilitate the antemortem diagnosis of C. felis infection. Clinical and pathological diagnosis of cytauxzoonosis is based on microscopic identification of parasites in tissues or on blood smears. We have developed and evaluated the sensitivity and specificity of a polymerase chain reaction (PCR) based assay for the diagnosis of C. felis infections in feline blood samples. The assay is sensitive enough to detect one copy of a cloned fragment of the C. felis 18S rRNA gene. This PCR assay can be used for the rapid clinical diagnosis of cytauxzoonosis and for epidemiological studies that will better define the geographic distribution of C. felis infection in cats.}, number={1-2}, journal={VETERINARY PARASITOLOGY}, author={Birkenheuer, AJ and Marr, H and Alleman, AR and Levy, MG and Breitschwerdt, EB}, year={2006}, month={Apr}, pages={144–149} }
 @article{gookin_copple_papich_poore_stauffer_birkenheuer_twedt_levy_2006, title={Efficacy of ronidazole for treatment of feline Tritrichomonas foetus infection}, volume={20}, ISSN={["1939-1676"]}, DOI={10.1892/0891-6640(2006)20[536:EORFTO]2.0.CO;2}, abstractNote={To determine the efficacy of ronidazole (RDZ), tinidazole (TDZ), and metronidazole (MDZ) against Tritrichomonas foetus in vitro and of RDZ for treatment of feline naturally occurring or experimentally induced T. foetus infection.A cat naturally infected with T. foetus infection and diarrhea. Ten specific-pathogen-free (SPF) kittens.RDZ, TDZ, and MDZ were tested for activity against 3 different feline isolates of T. foetus in vitro. RDZ then was administered to a naturally infected cat at 10 mg/kg PO q24h for 10 days. SPF kittens were infected orogastrically with feline T. foetus and treated with either placebo or RDZ (10 mg/kg PO q12h for 14 days). Cats with relapsing infection or those receiving placebo were treated subsequently with RDZ (either 30 or 50 mg/kg PO q12h for 14 days). Feces were examined for T. foetus by direct microscopy, culture, and polymerase chain reaction (PCR) testing weekly.Both RDZ and TDZ killed T. foetus at concentrations >0.1 microg/mL in vitro. In the naturally infected cat, RDZ abolished diarrhea and T. foetus infection for 85 days after treatment, at which time infection and diarrhea relapsed. Retreatment with RDZ eradicated diarrhea and T. foetus infection for over 407 days. In experimentally induced infection, RDZ at 10 mg/kg caused initial improvement, but infection relapsed in all 5 cats 2 to 20 weeks after treatment. At 30 or 50 mg/kg, 10/10 cats were negative for T. foetus infection for follow-up durations of 21 to 30 weeks after treatment.Oral administration of RDZ at 30 to 50 mg/kg q12h for 14 days resolved diarrhea and eradicated infection (on the basis of polymerase chain reaction [PCR] testing) in 1 naturally infected cat and 10 experimentally inoculated cats receiving a different isolate of T. foetus.}, number={3}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Gookin, Jody L. and Copple, Christina N. and Papich, Mark G. and Poore, Matthew F. and Stauffer, Stephen H. and Birkenheuer, Adam J. and Twedt, David C. and Levy, Michael G.}, year={2006}, pages={536–543} }
 @article{birkenheuer_whittington_neel_large_barger_levy_breitschwerdt_2006, title={Molecular characterization of a Babesia species identified in a North American raccoon}, volume={42}, ISSN={["1943-3700"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-33748280658&partnerID=MN8TOARS}, DOI={10.7589/0090-3558-42.2.375}, abstractNote={Piroplasmosis was first described in raccoons (Procyon lotor) in 1926, and the official description of a small piroplasm as Babesia lotori was done in 1981. Babesia microti-like gene sequences have been characterized in raccoons in both North American and Japan. It is well documented that the microscopic appearance of piroplasms does not always accurately predict the genotype and phylogenetic classification. Discrepancies using phenotype to predict genotype have been reported most frequently when evaluating small piroplasms. We amplified and sequenced the full-length 18S rRNA gene from a small piroplasm identified in a raccoon and used this sequence for phylogenetic analyses. Based on these analyses, the organism was placed in the Babesia sensu stricto clade, confirming that it is a true Babesia sp. This documents that at least two Babesia spp. can infect raccoons. The data generated in this study can be used to design molecular diagnostic tests for detection of this Babesia sp., which will be useful for epidemiologic and comparative phylogenetic studies. As piroplasmosis has been documented with increased frequency in humans in recent years, the results of this study will aid in the recognition of zoonotic babesiosis.}, number={2}, journal={JOURNAL OF WILDLIFE DISEASES}, author={Birkenheuer, Adam J. and Whittington, Julia and Neel, Jennifer and Large, Edward and Barger, Anne and Levy, Michael G. and Breitschwerdt, Edward B.}, year={2006}, month={Apr}, pages={375–380} }
 @article{schantz_steurer_duprey_kurpel_barr_jackson_breitschwerdt_levy_fox_2005, title={Autochthonous visceral leishmaniasis in dogs in North America}, volume={226}, ISSN={0003-1488}, url={http://dx.doi.org/10.2460/javma.2005.226.1316}, DOI={10.2460/javma.2005.226.1316}, number={8}, journal={Journal of the American Veterinary Medical Association}, publisher={American Veterinary Medical Association (AVMA)}, author={Schantz, Peter M. and Steurer, Francis J. and Duprey, Zandra H. and Kurpel, Katherine P. and Barr, Stephen C. and Jackson, Joan E. and Breitschwerdt, Edward B. and Levy, Michael G. and Fox, J. C.}, year={2005}, month={Apr}, pages={1316–1322} }
 @article{flowers_poore_pote_litaker_levy_2005, title={Cercariae of Bolbophorus damnificus and Bolbophorus sp with notes on North American bolbophorids}, volume={72}, ISSN={["1938-2952"]}, DOI={10.1654/4173}, abstractNote={Single-species and dual infections of Bolbophorus damnificus and a second cryptic species of Bolbophorus were distinguished in marsh rams-horn snails, Planorbella trivolvis, from aquaculture ponds in Mississippi, U.S.A. The cercariae of both B. damnificus and Bolbophorus sp. are described and distinguished using differences in body length, tail stem length, intestinal primordia, and integument spine patterns.}, number={2}, journal={COMPARATIVE PARASITOLOGY}, author={Flowers, JR and Poore, MF and Pote, LM and Litaker, RW and Levy, MG}, year={2005}, month={Jul}, pages={220–226} }
 @article{levy_noga_2005, title={Controlling parasitic dinoflagellates of fish, with special emphasis on molecular genetics and immunity}, volume={51}, ISBN={0001-7302}, number={4}, journal={Acta Zootaxonomica Sinica}, author={Levy, M. G. and Noga, E. J.}, year={2005}, pages={550} }
 @article{birkenheuer_correa_levy_breitschwerdt_2005, title={Geographic distribution of babesiosis among dogs in the United States and association with dog bites: 150 cases (2000-2003)}, volume={227}, ISSN={0003-1488}, url={http://dx.doi.org/10.2460/javma.2005.227.942}, DOI={10.2460/javma.2005.227.942}, abstractNote={To identify the geographic distribution of babesiosis among dogs in the United States and determine, for dogs other than American Pit Bull Terriers (APBTs), whether infection was associated with a recent dog bite.Retrospective study.150 dogs.Canine blood samples submitted to the North Carolina State University Vector-Borne Disease Diagnostic Laboratory between May 2000 and October 2003 for which results of a Babesia-specific polymerase chain reaction assay were positive were identified, and breed and geographic origin of dogs from which samples were obtained were recorded. History and hematologic abnormalities for dogs that were not APBTs were recorded, and possible associations with a recent dog bite were examined.Dogs positive for Babesia DNA were located in 29 states and 1 Canadian province (Ontario). Babesia gibsoni was the most commonly detected species, with B gibsoni DNA detected in blood samples from 131 of 144 (91%) dogs. Of the 131 dogs positive for B gibsoni DNA, 122 (93%) were APBTs. Of the 10 dogs positive for Babesia canis vogeli DNA, 6 were Greyhounds. In dogs other than APBTs, there was an association between having recently been bitten by another dog, particularly an APBT, and infection with B gibsoni.Results document an expansion of the known geographic range for babesiosis among dogs in the United States. Testing for babesiosis should be pursued in dogs with clinicopathologic abnormalities consistent with immune-mediated hemolytic anemia or thrombocytopenia, particularly if there is a history of a recent dog bite.}, number={6}, journal={Journal of the American Veterinary Medical Association}, publisher={American Veterinary Medical Association (AVMA)}, author={Birkenheuer, Adam J. and Correa, Maria T. and Levy, Michael G. and Breitschwerdt, Edward B.}, year={2005}, month={Sep}, pages={942–947} }
 @article{gookin_birkenheuer_st john_spector_levy_2005, title={Molecular characterization of trichomonads from feces of dogs with diarrhea}, volume={91}, ISSN={["1937-2345"]}, DOI={10.1645/ge-474r.1}, abstractNote={Trichomonads are occasionally observed in the feces of dogs with diarrhea. On the basis of superficial morphological appearance, these infections have been attributed to opportunistic overgrowth of the commensal, Pentatrichomonas hominis. However, molecular characterization of canine trichomonads has never been reported. This study was performed to determine, by means of rRNA gene sequence analysis, the identity of trichomonads observed in feces from dogs with diarrhea. Total DNA was isolated from fecal samples obtained from a 3-mo-old mixed breed dog and litter of German Shepherd puppies having profuse liquid diarrhea containing numerous trichomonads. Total DNA was subject to PCR amplification of partial 18S rRNA gene or 5.8S, ITS1, ITS2, and partial 18S and 28S rRNA genes using species-specific and universal primers, respectively. Products of 642 and 1864 base-pair length were amplified and cloned. On the basis of rRNA gene sequence, the trichomonads observed in the single dog and the litter of puppies shared 100% identity with Tritrichomonas foetus and P. hominis, respectively. The present study is the first to establish the molecular identity of trichomonads infecting dogs with diarrhea. These studies validate the longstanding assumption that canine trichomoniasis may be attributed to P. hominis. Importantly, these studies additionally recognize that canine trichomoniasis may also be caused by infection with T. foetus.}, number={4}, journal={JOURNAL OF PARASITOLOGY}, author={Gookin, JL and Birkenheuer, AJ and St John, V and Spector, M and Levy, MG}, year={2005}, month={Aug}, pages={939–943} }
 @article{bakal_hickson_gilger_levy_flowers_khoo_2005, title={Surgical Removal of Cataracts Due to Diplostomum Species in Gulf Sturgeon (Acipenser oxyrinchus desotoi)}, volume={36}, ISSN={1042-7260 1937-2825}, url={http://dx.doi.org/10.1638/04-044.1}, DOI={10.1638/04-044.1}, abstractNote={Twenty 6-yr-old (1995-yr-class) Gulf of Mexico sturgeon (Acipenser oxyrinchus desotoi) were diagnosed as having bilateral cataracts. Histopathologic assessment of the lenses of two of the fish revealed the presence of a diplostomid trematode. Pharmacological treatment of the trematodes may be effective for killing the parasites, but the damage to the lenses and resulting cataracts are nonreversible. Because these animals were to be used in a subsequent study as sentinels in the natural environment, it was necessary to return the animals' vision to as close to normal as possible. Electroretinograms were performed on each fish's eyes to ensure that retinal function was present. Cataracts then were surgically removed by phacoemulsification and aspiration. The animals tolerated the surgical procedures well. This report is the first known report of surgical correction of cataracts in sturgeon species. It also is the first known attempt to correct vision problems in fish being returned to the wild.}, number={3}, journal={Journal of Zoo and Wildlife Medicine}, publisher={American Association of Zoo Veterinarians}, author={Bakal, Robert S. and Hickson, Brian H. and Gilger, Brian C. and Levy, Michael G. and Flowers, James R. and Khoo, Lester}, year={2005}, month={Sep}, pages={504–508} }
 @article{dzikowski_levy_poore_flowers_paperna_2004, title={Clinostomum complanatum and Clinostomum marginatum (Rudolphi, 1819) (Digenea : Clinostomidae) are separate species based on differences in ribosomal DNA}, volume={90}, ISSN={["0022-3395"]}, DOI={10.1645/GE-159R}, abstractNote={Infections by metacercariae of Clinostomum (Leidy, 1856) species adversely affect aquacultured fish and are potentially transmissible to humans. Molecular methodologies are efficient tools, which enable diagnosis of all life-history stages of trematodes in their diverse hosts. The small subunit of ribosomal DNA genes of adults of the Old World Clinostomum complanatum (Rudolphi, 1819) and the New World Clinostomum marginatum (Rudolphi, 1819), obtained from a little egret Egretta garzetta (Linnaeus, 1766) and the great blue heron Ardea herodias (Linnaeus, 1758), respectively, were amplified, sequenced, and aligned. The resulting alignment was used to develop a genetic assay to differentiate between these species.}, number={2}, journal={JOURNAL OF PARASITOLOGY}, author={Dzikowski, R and Levy, MG and Poore, MF and Flowers, JR and Paperna, I}, year={2004}, month={Apr}, pages={413–414} }
 @article{birkenheuer_neel_ruslander_levy_breitschwerdt_2004, title={Detection and molecular characterization of a novel large Babesia species in a dog}, volume={124}, ISSN={0304-4017}, url={http://dx.doi.org/10.1016/j.vetpar.2004.07.008}, DOI={10.1016/j.vetpar.2004.07.008}, abstractNote={Babesia canis has generally been considered the only large Babesia to infect dogs. Here we describe the molecular characterization of a large Babesia species that was detected in the blood and bone marrow of a dog with clinical and hematological abnormalities consistent with babesiosis. Analysis of the 18S rRNA genes revealed a unique sequence that shared 93.9% sequence identity with B. bigemina and 93.5% sequence identity with B. caballi, compared to 91.2–91.6% identity with B. canis canis, B. c. vogeli, and B. c. rossi. Cross-reactive antibodies against B. canis, B. gibsoni (Asian genotype), or B. gibsoni (California genotype) antigens were not detected in acute or convalescent serum samples. The dog was treated with imidocarb diproprionate, which resulted in the resolution of clinical signs, and subsequently Babesia DNA was not detectable by PCR in post-treatment samples. The organism described in this report represents a genetically unique large Babesia sp. and is the eighth genetically distinct piroplasm capable of infecting the domestic dog.}, number={3-4}, journal={Veterinary Parasitology}, publisher={Elsevier BV}, author={Birkenheuer, A.J. and Neel, J. and Ruslander, D. and Levy, M.G. and Breitschwerdt, E.B.}, year={2004}, month={Oct}, pages={151–160} }
 @article{flowers_poore_mullen_levy_2004, title={Digeneans collected from piscivorous birds in north Carolina, USA}, volume={71}, ISSN={["1938-2952"]}, DOI={10.1654/4117}, abstractNote={During investigations of digenetic fluke diseases of aquaculture fish, 11 digeneans (Apharyngostrigea simplex, Apharyngostrigea cornu, Diplostomum compactum, Diplostomum spathaceum, Posthodiplostomum minimum, Hysteromorpha triloba, Clinostomum marginatum, Drepanocephalus spathans, Mesorchis denticulatus, Microparyphium facetum, and Notocotylus pacifera) were collected from 5 species of piscivorous birds (Ardea herodias, Fulica americana, Larus delawarensis, Nycticorax nycticorax, and Phalacrocorax auritus) from North Carolina, U.S.A. Apharyngostrigea simplex from A. herodias represents a new host record. Diplostomum spathaceum, P. minimum, C. marginatum, and M. denticulatus have previously been reported from North Carolina; the remainder represent new locality records.}, number={2}, journal={COMPARATIVE PARASITOLOGY}, author={Flowers, JR and Poore, MF and Mullen, JE and Levy, MG}, year={2004}, month={Jul}, pages={243–244} }
 @article{birkenheuer_levy_breitschwerdt_2004, title={Efficacy of combined atovaquone and azithromycin for therapy of chronic Babesia gibsoni (Asian genotype) infections in dogs}, volume={18}, ISSN={["1939-1676"]}, DOI={10.1892/0891-6640(2004)18<494:EOCAAA>2.0.CO;2}, abstractNote={Babesiosis caused by Babesia gibsoni (Asian genotype) is an emerging disease in dogs in the United States. To date, no drugs have been shown to eliminate B. gibsoni (Asian genotype) infections from dogs. Twenty-two dogs that remained persistently infected with B. gibsoni (Asian genotype) after either imidocarb diproprionate and or diminazine aceturate therapy were identified and randomly and evenly distributed into 2 groups. One group was treated with atovaquone and azithromycin combination therapy, and the other group received a placebo. Eight of 10 dogs in the treatment group had no detectable B. gibsoni (Asian genotype) DNA, as determined by a sensitive and specific polymerase chain reaction (PCR) assay, in any of their posttreatment samples. In contrast, B. gibsoni (Asian genotype) DNA was detectable by PCR in the posttreatment samples from 11 of 11 of the placebo-treated dogs. One dog in the treatment group was excluded from the treatment outcome analysis. This dog had 2 consecutive negative PCR assay results and was euthanized because of ongoing degenerative joint disease prior to completion of the study. No adverse effects of treatment were reported in any dog during the study period. A combination of atovaquone and azithromycin is the 1st described treatment that will either eliminate B. gibsoni (Asian genotype) infections or suppress the parasitemia below the limit of detection in the majority of treated dogs.}, number={4}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Birkenheuer, AJ and Levy, MG and Breitschwerdt, EB}, year={2004}, pages={494–498} }
 @article{foster_gookin_poore_stebbins_levy_2004, title={Outcome of cats with diarrhea and Tritrichomonas foetus infection}, volume={225}, ISSN={["1943-569X"]}, url={https://doi.org/10.2460/javma.2004.225.888}, DOI={10.2460/javma.2004.225.888}, abstractNote={To determine the long-term outcome of cats infected with Tritrichomonas foetus and identify treatment and management strategies influencing resolution of infection or associated diarrhea.Prospective study.26 cats with T. foetus-associated diarrhea at least 22 months prior to the study.A standardized survey regarding clinical course and management was administered to owners of cats with T. foetus infection and associated diarrhea. Fecal samples were obtained from each cat; the presence of T. foetus was assessed via microscopic examination of smears, culture in commercial media, and polymerase chain reaction amplification of T. foetus rDNA involving species-specific primers.Survey responses were obtained from owners of all 26 cats. Twenty-three cats had complete resolution of diarrhea a median of 9 months after onset. Analysis of fecal samples obtained from 22 cats revealed persistent T. foetus infection in 12, with a median of 39 months after resolution of diarrhea. History of implementation of a dietary change, treatment with paromomycin, or higher numbers of cats in the household was associated with significantly longer duration of time to resolution of diarrhea.Results suggested chronic T. foetus-associated diarrhea in most cats is likely to resolve spontaneously within 2 years of onset. Chronic infection with T. foetus (without clinical signs) after resolution of diarrhea appears to be common. Although often temporarily effective in decreasing severity of diarrhea, attempts to treat cats with T. foetus infection may result in prolongation of time to resolution of diarrhea.}, number={6}, journal={JAVMA-JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION}, author={Foster, DM and Gookin, JL and Poore, MF and Stebbins, ME and Levy, MG}, year={2004}, month={Sep}, pages={888–892} }
 @article{gookin_stebbins_hunt_burlone_fulton_hochel_talaat_poore_levy_2004, title={Prevalence of and risk factors for feline Tritichomonas foetus and Giardia infection}, volume={42}, ISSN={["1098-660X"]}, DOI={10.1128/JCM.42.6.2707-2710.2004}, abstractNote={ABSTRACT Data were gathered for 117 cats from 89 catteries at an international cat show to examine prevalence and risk factors for feline Tritrichomonas foetus and Giardia infection. Prevalence of T. foetus was 31% among cats (36 out of 117) and catteries (28 out of 89) based on results of fecal smear examination (5 out of 36), fecal culture in modified Diamond's medium (9 out of 36), fecal culture in In Pouch TF medium (20 out of 36), or PCR amplification of the ribosomal RNA gene from feces with T. foetus -specific primers (34 out of 36). Catteries in which T. foetus was identified were more likely to have had a recent history of diarrhea, historical diagnosis of coccidia infection in adult cats, and a decreased number of square feet of facility per cat. Evidence did not exist for the ongoing transmission of T. foetus by water, food, or contact with other species.}, number={6}, journal={JOURNAL OF CLINICAL MICROBIOLOGY}, author={Gookin, JL and Stebbins, ME and Hunt, E and Burlone, K and Fulton, M and Hochel, R and Talaat, M and Poore, M and Levy, MG}, year={2004}, month={Jun}, pages={2707–2710} }
 @article{dzikowski_levy_poore_flowers_paperna_2004, title={Use of rDNA polymorphism for identification of heterophyidae infecting freshwater fishes}, volume={59}, ISSN={["1616-1580"]}, DOI={10.3354/dao059035}, abstractNote={DAO Diseases of Aquatic Organisms Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials DAO 59:35-41 (2004) - doi:10.3354/dao059035 Use of rDNA polymorphism for identification of Heterophyidae infecting freshwater fishes R. Dzikowski1,*, M. G. Levy2, M. F. Poore2, J. R. Flowers2, I. Paperna1 1 Department of Animal Sciences, Faculty of Agriculture, Food and Environmental Sciences, Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel 2 Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 4700 Hillsborough St., Raleigh 27606, North Carolina, USA *Email: rrd2001@med.cornell.edu ABSTRACT: Infections by trematodes are among the most common fish-borne zoonoses. Metacercariae of the Family Heterophyidae in marine and freshwater fishes are nonfastidious in their choice of definitive hosts, and therefore, cause infections in human and domestic animals. In the present study, species-specific polymerase chain reaction (PCR) assays were developed for identifying and differentiating the various species examined. Sequencing and aligning the 18S (SSU) rDNA revealed interspecific variation for which species-specific DNA oligonucleotides were designed and used for the identification of 6 heterophyid species recovered from piscivorous birds. The oligonucleotides were further used to evaluate the various stages (cercariae recovered from snails, metacercariae recovered from fish and adult trematodes) of the digeneans. By applying this method we elucidated for the first time the life cycle of Pygidiopsis genata. The phylogenetic interrelationship among the newly sequenced species of Heterophyidae is outlined. KEY WORDS: Digenea · Heterophyidae · Birds · Fish · 18S rDNA gene · SSU Full article in pdf format PreviousNextExport citation RSS - Facebook - Tweet - linkedIn Cited by Published in DAO Vol. 59, No. 1. Online publication date: April 21, 2004 Print ISSN: 0177-5103; Online ISSN: 1616-1580 Copyright © 2004 Inter-Research.}, number={1}, journal={DISEASES OF AQUATIC ORGANISMS}, author={Dzikowski, R and Levy, MG and Poore, MF and Flowers, JR and Paperna, I}, year={2004}, month={Apr}, pages={35–41} }
 @article{tuttle_birkenheuer_juopperi_levy_breitschwerdt_2003, title={Concurrent bartonellosis and babesiosis in a dog with persistent thrombocytopenia}, volume={223}, ISSN={0003-1488}, url={http://dx.doi.org/10.2460/javma.2003.223.1306}, DOI={10.2460/javma.2003.223.1306}, abstractNote={A 12-year-old castrated male West Highland White Terrier was referred because of recurrent episodes of collapsing. The dog was mildly anemic and severely thrombocytopenic and had high serum alanine aminotransferase activity. Infection with Bartonella vinsonii (berkhoffii) was initially diagnosed on the basis of serologic testing. Despite treatment with a series of antimicrobials and prolonged use of immunosuppressive drugs, thrombocytopenia persisted. After 5 months of treatment, Babesia canis organisms were seen during examination of a direct blood smear. The dog was treated with imidocarb dipropionate for babesiosis, after which thrombocytopenia resolved, and administration of immunosuppressive drugs was discontinued. Retrospective review of blood smears failed to identify organisms; however, polymerase chain reaction (PCR) analysis of multiple stored blood samples obtained during the 5-month period of persistent thrombocytopenia identified DNA of B. canis vogeli. Babesiosis may cause persistent, unexplained thrombocytopenia in dogs that are not anemic. A PCR assay can facilitate a diagnosis of babesiosis when organisms are not evident or when serologic testing fails to detect Babesia-specific antibodies.}, number={9}, journal={Journal of the American Veterinary Medical Association}, publisher={American Veterinary Medical Association (AVMA)}, author={Tuttle, Allison D. and Birkenheuer, Adam J. and Juopperi, Tarja and Levy, Michael G. and Breitschwerdt, Edward B.}, year={2003}, month={Nov}, pages={1306–1310} }
 @article{birkenheuer_levy_breitschwerdt_2003, title={Development and Evaluation of a Seminested PCR for Detection and Differentiation of Babesia gibsoni (Asian Genotype) and B. canis DNA in Canine Blood Samples}, volume={41}, ISSN={0095-1137}, url={http://dx.doi.org/10.1128/jcm.41.9.4172-4177.2003}, DOI={10.1128/JCM.41.9.4172-4177.2003}, abstractNote={ABSTRACT Canine babesiosis has recently been recognized as an emerging infectious disease of dogs in North America. We sought to develop a seminested PCR to detect and differentiate Babesia gibsoni (Asian genotype), B. canis subsp. vogeli , B. canis subsp. canis , and B. canis subsp. rossi DNA in canine blood samples. An outer primer pair was designed to amplify an ∼340-bp fragment of the 18S rRNA genes from B. gibsoni (Asian genotype), B. canis subsp. vogeli , B. canis subsp. rossi , and B. canis subsp. canis but not mammalian DNA. Forward primers were designed that would specifically amplify a smaller fragment from each organism in a seminested PCR. The practical limit of detection was 50 organisms/ml of mock-infected EDTA anticoagulated whole blood. The primer pair also amplified an ∼370-bp fragment of the B. gibsoni (USA/California genotype) 18S rRNA gene from the blood of an experimentally infected dog with a high percentage of parasitemia. Amplicons were not detected when DNA extracted from the blood of a dog that was naturally infected with Theileria annae at a low percentage of parasitemia was amplified. Due to limited sensitivity, this test is not recommended for the routine diagnosis of B. gibsoni (USA/California genotype) or T. annae . The PCR test did not amplify Toxoplasma gondii , Neospora caninum , Leishmania infantum , Cryptosporidium parvum , or canine DNA under any of the conditions tested. The seminested PCR test was able to detect and discriminate B. gibsoni (Asian genotype), B. canis subsp. vogeli , B. canis subsp. canis , and B. canis subsp. rossi DNA in blood samples from infected dogs.}, number={9}, journal={Journal of Clinical Microbiology}, publisher={American Society for Microbiology}, author={Birkenheuer, A. J. and Levy, M. G. and Breitschwerdt, E. B.}, year={2003}, month={Sep}, pages={4172–4177} }
 @article{dzikowski_levy_poore_flowers_paperna_2003, title={Genetic and morphologic differentiation of Bolbophorus confusus and B-levantinus (Digenea : Diplostomatidae), based on rDNA SSU polymorphism and SEM}, volume={57}, ISSN={["1616-1580"]}, DOI={10.3354/dao057231}, abstractNote={Metacercariae of Bolbophorus species are serious pathogens of farmed fish. Molecular diagnostic tools, capable of identifying and differentiating these parasites, may assist in the development of rationale control strategies. The rDNA 18S (small sub-unit: SSU) genes of adult B. confusus and B. levantinus obtained from a pelican, Pelecanus onocrotalus, and a night heron, Nycticorax nycticorax, respectively, were amplified, sequenced, and aligned. Based on this alignment, we developed a genetic differentiation assay between B. confusus and B. levantinus. These 2 species were compared genetically with the North American species B. damnificus and Bolbophorus sp. ('Type 2'). The relationship between species is outlined and discussed. In addition to the molecular study, specimens of B. confusus and B. levantinus were compared morphologically, using scanning electron microscopy. Morphologic analysis revealed interspecific differences in details of the holdfast organ and the position of the acetabulum.}, number={3}, journal={DISEASES OF AQUATIC ORGANISMS}, author={Dzikowski, R and Levy, MG and Poore, MF and Flowers, JR and Paperna, I}, year={2003}, month={Dec}, pages={231–235} }
 @article{birkenheuer_levy_stebbins_poore_breitschwerdt_2003, title={Serosurvey of antiBabesia antibodies in stray dogs and American pit bull terriers and American Staffordshire terriers from North Carolina}, volume={39}, ISSN={["0587-2871"]}, DOI={10.5326/0390551}, abstractNote={Stray dogs (n=359) and kennel dogs (n=149) from North Carolina were tested for evidence of antiBabesia antibodies. AntiBabesia antibodies were detected in 21/359 and 22/149 of the stray and kennel dogs, respectively. A total of 57 dogs from both groups were tested for babesiasis by light microscopy and polymerase chain reaction (PCR). Babesia deoxyribonucleic acid (DNA) was detected in 3/28 of the stray dogs and 14/29 of the kennel dogs. When Babesia DNA was detected by PCR, the species-specific PCR results differed from the Babesia species antibody titer results in 6/17 of the PCR-positive dogs. There was no association between antiBabesia antibodies and the presence of ticks. There are currently Babesia gibsoni epizootics affecting American pit bull terrier kennels.}, number={6}, journal={JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION}, author={Birkenheuer, AJ and Levy, MG and Stebbins, M and Poore, M and Breitschwerdt, E}, year={2003}, pages={551–557} }
 @article{levy_gookin_poore_birkenheuer_dykstra_litaker_2003, title={Tritrichomonas foetus and not Pentatrichomonas hominis is the etiologic agent of feline trichomonal diarrhea}, volume={89}, ISSN={["1937-2345"]}, DOI={10.1645/0022-3395(2003)089[0099:TFANPH]2.0.CO;2}, abstractNote={Recently, several investigators have reported large-bowel diarrhea in cats associated with intestinal trichomonad parasites. These reports have presumptively identified the flagellates as Pentatrichomonas hominis, a n organism putatively capable of infecting the intestinal tracts of a number of mammalian hosts, including cats, dogs, and man. The purpose of the present study was to determine the identity of this recently recognized flagellate by means of rRNA gene sequence analysis; restriction enzyme digest mapping; and light, transmission, and scanning electron microscopy (SEM).}, number={1}, journal={JOURNAL OF PARASITOLOGY}, author={Levy, MG and Gookin, JL and Poore, M and Birkenheuer, AJ and Dykstra, MJ and Litaker, RW}, year={2003}, month={Feb}, pages={99–104} }
 @article{gookin_foster_poore_stebbins_levy_2003, title={Use of a commercially available culture system for diagnosis of Tritrichomonas foetus infection in cats}, volume={222}, url={https://doi.org/10.2460/javma.2003.222.1376}, DOI={10.2460/javma.2003.222.1376}, abstractNote={To evaluate the efficacy of and optimize a commercially available culture system for sensitive and specific in-clinic culture of Tritrichomonas foetus from cat feces.Prospective study.Samples of freshly voided feces from 117 purebred cats and pure cultures of T. foetus obtained from a cat with chronic diarrhea.Optimal conditions for use of the culture system, such as quantity of fecal inoculum (0.025 to 0.2 g) and cultivation temperature (25 or 37 degrees C [98.6 or 77.0 degrees F]), were determined. Specificity of the system was examined by attempted culture of Giardia lamblia and Pentatrichomonas hominis. Sensitivity of the system to detect T. foetus was determined by inoculation of culture system pouches with serially diluted T. foetus suspensions with and without feces.Detection limit of the culture system was 1 and 1,000 T. foetus organisms without and with feces from cats, respectively. Optimal fecal inoculum was < 0.1 g of feces. At 37 degrees C, cultures yielded positive results in 24 hours; organisms remained viable for 1 to 6 days, and bacterial overgrowth was common. At 25 degrees C, cultures yielded positive results in 1 to 11 days; organisms were long-lived, and bacterial overgrowth was uncommon. Neither G. lamblia or P. hominis survived in the culture system.The culture system was sensitive and specific for culture of T. foetus in feces of cats. Performance was optimal when test kits were inoculated with < or = 0.1 g of freshly voided feces and cultured at 25 degrees C.}, number={10}, journal={JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION}, author={Gookin, JL and Foster, DM and Poore, MF and Stebbins, ME and Levy, MG}, year={2003}, month={May}, pages={1376–1379} }
 @article{flowers_hammerberg_wood_malarkey_dam_levy_mclawhorn_2002, title={Heterobilharzia americana infection in a dog}, volume={220}, ISSN={["0003-1488"]}, DOI={10.2460/javma.2002.220.193}, abstractNote={A 7-year-old castrated male Golden Retriever cross was evaluated because of intermittent blood-tinged diarrhea, severe weight loss, anorexia, and lethargy of 2 months' duration; the dog was unresponsive to antimicrobial and standard anthelmintic treatment. Results of fecal flotations for parasite ova were negative. Alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase activities and total protein and globulin conentrations were greater than reference ranges. Biopsy specimens were obtained during laparotomy and examination revealed multiple granulomatous lesions with helminth ova nidi in the intestine, pancreas, liver, and mesenteric lymph node. Saline solution direct smear and saline solution sedimentation of feces yielded trematode ova that were morphologically consistent with Heterobilharzia americana. Identification was confirmed when miracidia were hatched from these ova and produced characteristic cercariae from infected snails. An antigen capture ELISA, typically used for the diagnosis of schistosomiasis in humans, was performed, and schistosome circulating anodic antigen was detected. Treatment with 30 mg of praziquantel/kg (14 mg/lb) of body weight stopped ova shedding, removed detectable circulating antigens, and caused the dog's body weight and attitude to return to normal. Although this is the first report of canine heterobilharziasis in North Carolina, it suggests that heterobilharziasis is underdiagnosed in dogs that have contact with water frequented by raccoons. Inappropriate diagnostic procedures can foil accurate detection of this parasitic disease.}, number={2}, journal={JOURNAL OF THE AMERICAN VETERINARY MEDICAL ASSOCIATION}, author={Flowers, JR and Hammerberg, B and Wood, SL and Malarkey, DE and Dam, GJ and Levy, MG and McLawhorn, LD}, year={2002}, month={Jan}, pages={193–196} }
 @article{levy_flowers_poore_mullen_khoo_pote_paperna_dzikowski_litaker_2002, title={Morphologic, pathologic, and genetic investigations of Bolbophorus species affecting cultured channel catfish in the Mississippi delta}, volume={14}, ISSN={["0899-7659"]}, DOI={10.1577/1548-8667(2002)014<0235:MPAGIO>2.0.CO;2}, abstractNote={Trematodes belonging to the genus Bolbophorus have recently been reported as the cause of substantial morbidity and mortality in cultured channel catfish Ictalurus punctatus in Mississippi and Louisiana. Previous investigators identified only a single species, B. confusus. In this investigation, genetic techniques were used to identify all stages of the parasite in all of its hosts. The 18s rRNA genes from specimens collected in Mississippi were sequenced and compared; this analysis revealed that there are two distinct species, B. damnificus (previously identified as B. confusus) and another, undescribed species. (Phylogenetic analysis indicated that a third species, B. levantinus, is also closely related to the Mississippi species.) Species-specific polymerase chain reaction assays capable of identifying and differentiating between these two parasites were developed. Both species were found to infect the first intermediate host (the ram's horn snail Planorbella trivolvis) in commercial channel catfish ponds, but only B. damnificus was recovered from the fish themselves. The new, unidentified Bolbophorus species was determined to be highly pathogenic to a number of fish species. The contribution of B. damnificus to disease in cultured channel catfish remains undetermined. Future investigations of these parasites must now take into account the presence of two distinct species.}, number={4}, journal={JOURNAL OF AQUATIC ANIMAL HEALTH}, author={Levy, MG and Flowers, JR and Poore, MF and Mullen, JE and Khoo, LH and Pote, LM and Paperna, I and Dzikowski, R and Litaker, RW}, year={2002}, month={Dec}, pages={235–246} }
 @article{gookin_birkenheuer_breitschwerdt_levy_2002, title={Single-Tube Nested PCR for Detection of Tritrichomonasfoetus in Feline Feces}, volume={40}, ISSN={0095-1137}, url={http://dx.doi.org/10.1128/jcm.40.11.4126-4130.2002}, DOI={10.1128/JCM.40.11.4126-4130.2002}, abstractNote={ABSTRACT Tritrichomonas foetus , a venereal pathogen of cattle, was recently identified as an inhabitant of the large intestine in young domestic cats with chronic diarrhea. Recognition of the infection in cats has been mired by unfamiliarity with T . foetus in cats as well as misdiagnosis of the organisms as Pentatrichomonas hominis or Giardia sp. when visualized by light microscopy. The diagnosis of T . foetus presently depends on the demonstration of live organisms by direct microscopic examination of fresh feces or by fecal culturing. As T . foetus organisms are fastidious and fragile, routine flotation techniques and delayed examination and refrigeration of feces are anticipated to preclude the diagnosis in numerous cases. The objective of this study was to develop a sensitive and specific PCR test for the diagnosis of feline T . foetus infection. A single-tube nested PCR was designed and optimized for the detection of T . foetus in feline feces by using a combination of novel (TFITS-F and TFITS-R) and previously described (TFR3 and TFR4) primers. The PCR is based on the amplification of a conserved portion of the T . foetus internal transcribed spacer (ITS) region (ITS1 and ITS2) and the 5.8S rRNA gene. The absolute detection limit of the single-tube nested PCR was 1 organism, while the practical detection limit was 10 organisms per 200 mg of feces. Specificity was examined by using P . hominis , Giardia lamblia, and feline genomic DNA. Our results demonstrate that the single-tube nested PCR is ideally suited for (i) diagnostic testing of feline fecal samples that are found negative by direct microscopy and culturing and (ii) definitive identification of microscopically observable or cultivated organisms.}, number={11}, journal={Journal of Clinical Microbiology}, publisher={American Society for Microbiology}, author={Gookin, J. L. and Birkenheuer, A. J. and Breitschwerdt, E. B. and Levy, M. G.}, year={2002}, month={Nov}, pages={4126–4130} }
 @article{gaskin_schantz_jackson_birkenheuer_tomlinson_gramiccia_levy_steurer_kollmar_hegarty_et al._2002, title={Visceral leishmaniasis in a New York foxhound kennel}, volume={16}, ISSN={["0891-6640"]}, DOI={10.1892/0891-6640(2002)016<0034:VLIANY>2.3.CO;2}, abstractNote={Although endemic throughout much of the world, autochthonous visceral leishmaniasis has been reported on only 3 previous occasions in North America. After diagnosis of visceral leishmaniasis in 4 foxhounds from a kennel in Dutchess County, New York (index kennel), serum and ethylenediamine-tetraacetic acid (EDTA)-anticoagulated blood were collected from the remaining 108 American or cross-bred foxhounds in the index kennel and from 30 Beagles and Basset Hounds that were periodically housed in the index kennel. Samples were analyzed for antibodies to or DNA of tickborne disease pathogens and Leishmania spp. Most dogs had antibodies to Rickettsia spp., Ehrlichia spp., Babesia spp., or some combination of these pathogens but not to Bartonella vinsonii (berkhoffi). However, DNA of rickettsial, ehrlichial, or babesial agents was detected in only 9 dogs. Visceral leishmaniasis was diagnosed in 46 of 112 (41%) foxhounds from the index kennel but was not diagnosed in any of the Beagles and Basset Hounds. A positive Leishmania status was defined by 1 or more of the following criteria: a Leishmania antibody titer > or = 1:64, positive Leishmania polymerase chain reaction (PCR), positive Leishmania culture, or identification of Leishmania amastigotes by cytology or histopathology. The species and zymodeme of Leishmania that infected the foxhounds was determined to be Leishmania infantum MON-1 by isoenzyme electrophoresis. Foxhounds that were > 18 months of age or that had traveled to the southeastern United States were more likely to be diagnosed with visceral leishmaniasis. Transmission of Leishmania spp. in kennel outbreaks may involve exposure to an insect vector, direct transmission, or vertical transmission.}, number={1}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Gaskin, AA and Schantz, P and Jackson, J and Birkenheuer, A and Tomlinson, L and Gramiccia, M and Levy, M and Steurer, F and Kollmar, E and Hegarty, BC and et al.}, year={2002}, pages={34–44} }
 @article{gookin_levy_law_papich_poore_breitschwerdt_2001, title={Experimental infection of cats with Tritrichomonas foetus}, volume={62}, ISSN={0002-9645}, url={http://dx.doi.org/10.2460/ajvr.2001.62.1690}, DOI={10.2460/ajvr.2001.62.1690}, abstractNote={To determine whether infection with Tritrichomonas foetus causes diarrhea in specific-pathogen-free or Cryptosporidium coinfected cats.4 cats with subclinical cryptosporidiosis (group 1) and 4 specific-pathogen-free cats (group 2).Cats were infected orogastrically with an axenic culture of T. foetus isolated from a kitten with diarrhea. Direct microscopy and protozoal culture of feces, fecal character, serial colonic mucosal biopsy specimens, and response to treatment with nitazoxanide (NTZ; group 1) or prednisolone (groups 1 and 2) were assessed.Infection with T. foetus persisted in all cats for the entire 203-day study and resulted in diarrhea that resolved after 7 weeks. Group-1 cats had an earlier onset, more severe diarrhea, and increased number of trichomonads on direct fecal examination, compared with group-2 cats. Use of NTZ eliminated shedding of T. foetus and Cryptosporidium oocysts, but diarrhea consisting of trichomonad-containing feces recurred when treatment was discontinued. Prednisolone did not have an effect on infection with T. foetus but resulted in reappearance of Cryptosporidium oocysts in the feces of 2 of 4 cats. During necropsy, T. foetus was isolated from contents of the ileum, cecum, and colon. Tritrichomonas foetus organisms and antigen were detected on surface epithelia and within superficial detritus of the cecal and colonic mucosa.After experimental inoculation in cats, T. foetus organisms colonize the ileum, cecum, and colon, reside in close contact with the epithelium, and are associated with transient diarrhea that is exacerbated by coexisting cryptosporidiosis but not treatment with prednisolone.}, number={11}, journal={American Journal of Veterinary Research}, publisher={American Veterinary Medical Association (AVMA)}, author={Gookin, Jody L. and Levy, Michael G. and Law, J. Mac and Papich, Mark G. and Poore, Matthew F. and Breitschwerdt, Edward B.}, year={2001}, month={Nov}, pages={1690–1697} }
 @article{birkenheuer_levy_savary_gager_breitschwerdt_1999, title={Babesia gibsoni infections in dogs from North Carolina}, volume={35}, ISSN={["0587-2871"]}, DOI={10.5326/15473317-35-2-125}, abstractNote={The recognition of canine babesiosis in North Carolina caused by Babesia gibsoni documents the expansion of the previously reported endemic area of this disease. Clinical signs ranged from severe hemolytic anemia and thrombocytopenia to subclinical infections. No infected dogs had traveled to endemic areas. Antibabesial treatment failed to eradicate the organism from infected dogs.}, number={2}, journal={JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION}, author={Birkenheuer, AJ and Levy, MG and Savary, KCM and Gager, RB and Breitschwerdt, EB}, year={1999}, pages={125–128} }
 @article{litaker_tester_colorni_levy_noga_1999, title={The phylogenetic relationship of Pfiesteria piscicida, Cryptoperidiniopsoid sp Amyloodinoum ocellatum and a Pfiesteria-like dinoflagellate to other dinoflagellates and apicomplexans}, volume={35}, ISSN={["1529-8817"]}, DOI={10.1046/j.1529-8817.1999.3561379.x}, abstractNote={The taxonomic relationship between heterotrophic and parasitic dinoflagellates has not been studied extensively at the molecular level. In order to investigate these taxonomic relationships, we sequenced the small subunit (SSU) ribosomal RNA gene of Pfiesteria piscicida (Steidinger et Burkholder), a Pfiesteria-like dinoflagellate, Cryptoperidiniopsoid sp., and Amyloodinium ocellatum (Brown) and submitted those sequences to GenBank. Pfiesteria piscicida and Cryptoperidiniopsoid sp. are heterotrophic dinoflagellates, purportedly pathogenic to fish, and A. ocellatum, a major fish pathogen, has caused extensive economic losses in both the aquarium and aquaculture industries. The pathogenicity of the Pfiesteria-like dinoflagellate is unknown at this time, but its growth characteristics and in vitro food preferences are similar to those of P. piscicda. The SSU sequences of these species were aligned with the other full-length dinoflagellate sequences, as well as those of representative apicomplexans and Perkinsus species, the groups most closely related to dinoflagellates. Phylogenetic analyses indicate that Cryptoperidiniopsoid sp., P. piscicida, and the Pfiesteria-like dinoflagellate are closely related and group into the class Blastodiniphyceae, as does A. ocellatum. None of the species examined were closely related to the apicomplexans or to Perkinsus marinus, the parasite that causes “Dermo disease” in oysters. The overall phylogenetic analyses largely supported the current class and subclass groupings within the dinoflagellates.}, number={6}, journal={JOURNAL OF PHYCOLOGY}, author={Litaker, RW and Tester, PA and Colorni, A and Levy, MG and Noga, EJ}, year={1999}, month={Dec}, pages={1379–1389} }
 @article{chae_levy_hunt_schlater_snider_waghela_holman_wagner_1999, title={Theileria sp infections associated with bovine fatalities in the United States confirmed by small-subunit rRNA gene analyses of blood and tick samples}, volume={37}, number={9}, journal={Journal of Clinical Microbiology}, author={Chae, J. S. and Levy, M. and Hunt, J. and Schlater, J. and Snider, G. and Waghela, S. D. and Holman, P. J. and Wagner, G. G.}, year={1999}, pages={3037–3040} }
 @article{cobb_levy_noga_1998, title={Acquired immunity to amyloodiniosis is associated with an antibody response}, volume={34}, ISSN={["1616-1580"]}, DOI={10.3354/dao034125}, abstractNote={DAO Diseases of Aquatic Organisms Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials DAO 34:125-133 (1998) - doi:10.3354/dao034125 Acquired immunity to amyloodiniosis is associated with an antibody response Charles S. Cobb, Michael G. Levy1,*, Edward J. Noga2 Departments of 1Microbiology, Pathology, and Parasitology, and 2Companion Animal and Special Species Medicine, North Carolina State University College of Veterinary Medicine, Raleigh, North Carolina 27606, USA *Addressee for correspondence. E-mail: mike_levy@ncsu.edu ABSTRACT: The dinoflagellate Amyloodinium ocellatum, which causes amyloodiniosis or 'marine velvet disease', is one of the most serious ectoparasitic diseases plaguing warmwater marine fish culture worldwide. We report that tomato clownfish Amphiprion frenatus develop strong immunity to Amyloodinium ocellatum infection following repeated nonlethal challenges and that specific antibodies are associated with this response. Reaction of immune fish antisera against dinospore and trophont-derived antigens in Western blots indicated both shared and stage-specific antibody-antigen reactions. A mannan-binding-protein affinity column was used to isolate IgM-like antibody from A. frenatus serum. The reduced Ig consisted of one 70 kD heavy chain and one 32 kD light chain with an estimated molecular weight of 816 kD for the native molecule. mmunoglobulin (Ig) isolated from immune but not non-immune fish serum significantly inhibited parasite infectivity in vitro. An enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal rabbit antibody produced against affinity-purified A. frenatus Ig. Anti-Amyloodinium serum antibody was not always detectable in immune fish, although serum antibody titers in immune fish increased after repeated exposure to the parasite. These results suggest that there may be a localized antibody response in skin/gill epithelial tissue, although antibody was rarely detected in skin mucus. KEY WORDS: Amyloodinium ocellatum · Immunity · Amphiprion frenatus · Ig Full text in pdf format PreviousNextExport citation RSS - Facebook - Tweet - linkedIn Cited by Published in DAO Vol. 34, No. 2. Publication date: October 08, 1998 Print ISSN:0177-5103; Online ISSN:1616-1580 Copyright © 1998 Inter-Research.}, number={2}, journal={DISEASES OF AQUATIC ORGANISMS}, author={Cobb, CS and Levy, MG and Noga, EJ}, year={1998}, month={Oct}, pages={125–133} }
 @article{robinette_wada_arroll_levy_miller_noga_1998, title={Antimicrobial activity in the skin of the channel catfish Ictalurus punctatus: characterization of broad-spectrum histone-like antimicrobial proteins}, volume={54}, ISSN={["1420-9071"]}, DOI={10.1007/s000180050175}, number={5}, journal={CELLULAR AND MOLECULAR LIFE SCIENCES}, author={Robinette, D and Wada, S and Arroll, T and Levy, MG and Miller, WL and Noga, EJ}, year={1998}, month={May}, pages={467–475} }
 @article{cobb_levy_noga_1998, title={Development of immunity by the tomato clownfish Amphiprion frenatus to the dinoflagellate parasite Amyloodinium ocellatum}, volume={10}, ISSN={["1548-8667"]}, DOI={10.1577/1548-8667(1998)010<0259:DOIBTT>2.0.CO;2}, abstractNote={The dinoflagellate Amyloodinium ocellatum, which causes amyloodiniosis or “marine velvet disease,” is one of the most serious ectoparasitic diseases affecting warmwater marine fish culture worldwide. We demonstrated that tomato clownfish Amphiprion frenatus can develop strong immunity to infection following repeated nonlethal parasitic challenges. The protective response is long-lived and directed against the trophont stage of the parasite.}, number={3}, journal={JOURNAL OF AQUATIC ANIMAL HEALTH}, author={Cobb, CS and Levy, MG and Noga, EJ}, year={1998}, month={Sep}, pages={259–263} }
 @article{wang_noga_avtalion_levy_1998, title={Whole blood assay for examining lymphocyte blastogenesis of percichthyid bass (Morone): Erratum}, volume={62}, number={4}, journal={Veterinary Immunology and Immunopathology}, author={Wang, C. J. and Noga, E. J. and Avtalion, R. and Levy, M. G.}, year={1998}, pages={367} }
 @article{hegarty_levy_gager_breitschwerdt_1997, title={Immunoblot Analysis of the Immunoglobulin G Response to Ehrlichia Canis in Dogs: An International Survey}, volume={9}, ISSN={1040-6387 1943-4936}, url={http://dx.doi.org/10.1177/104063879700900106}, DOI={10.1177/104063879700900106}, abstractNote={Historically, considerable variation has been reported in the type and severity of clinical and hematologic abnormalities associated with canine ehrlichiosis. Because of difficulties associated with the isolation of intracellular monocytic Ehrlichia species in tissue culture systems, few E. canis isolates are available for comparative microbiologic studies. To address the issue of potential E. canis antigenic diversity in different regions of the world, dog sera reactive by indirect fluorescent antibody testing to E. canis (Florida) antigen were obtained from France, Israel, Italy, the United States, the Virgin Islands, and Zimbabwe. Ehrlichia canis proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and at least 5 sera from each region were stained by western immunoblotting. Antibody immunodominance was scored based upon staining intensity. There was relative homogeneity in the immunogenic protein reactions to E. canis antigens. Of the 58 E. canis reactive sera, 54 samples resulted in immunoblot patterns indicative of chronic ehrlichiosis. Four reactive sera (reciprocal titers of 160–2,560) did not recognize any genus-specific antigens resulting in protein bands between 22 and 29 kD, indicating serologic cross-reactivity with other microorganisms. Relatively homogenous immunoblot patterns, consistent with the reported immunoblot response of dogs with experimental chronic ehrlichiosis, were observed with sera from Arizona, France, Israel, North Carolina, Texas, and the Virgin Islands. In contrast, unique major proteins were observed in dog sera from Italy and Zimbabwe. Our results indicate that although relatively homogeneous, antigenic diversity may exist among E. canis organisms in different regions of the world.}, number={1}, journal={Journal of Veterinary Diagnostic Investigation}, publisher={SAGE Publications}, author={Hegarty, Barbara C. and Levy, Michael G. and Gager, Robin F. and Breitschwerdt, Edward B.}, year={1997}, month={Jan}, pages={32–38} }
 @article{wang_noga_avtalion_levy_1997, title={Whole blood assay for examining lymphocyte blastogenesis of percichthyid bass (Morone) mrr01}, volume={58}, ISSN={["0165-2427"]}, DOI={10.1016/S0165-2427(97)00049-4}, abstractNote={A simple and reproducible method was developed for the measurement of blastogenesis of peripheral blood lymphocytes using whole blood of hybrid bass (striped bass [Morone saxatilis] female × white bass [M. chrysops] male) stimulated with Concanavalin A, phytohemagglutinin-P, lipopolysaccharide or pokeweed mitogen. Compared to traditional methods which use leucocyte separation procedures, whole blood culture is faster and less expensive. Only small aliquots of blood (10 μl per culture well) were needed, which would be beneficial for sampling small fish as well as for taking multiple samples from single animals. Optimal culture conditions for hybrid bass, including mitogen concentration, incubation temperature and incubation period, were determined. This is the first report to demonstrate a blastogenic response of whole blood cells in fish.}, number={3-4}, journal={VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY}, author={Wang, CJ and Noga, EJ and Avtalion, R and Levy, MG}, year={1997}, month={Sep}, pages={355–362} }
 @article{levine_levy_walker_crittenden_1988, title={An episode of cryptosporidiosis in veterinary students}, volume={193}, journal={Journal of the American Veterinary Medical Association}, author={Levine, J. F. and Levy, M. and Walker, R. L. and Crittenden, S.}, year={1988}, pages={1413–1414} }