Michael G Levy

Publications of Michael G Levy

• Experimental infection by capillary tube feeding of Rhipicephalus sanguineus with Bartonella vinsonii subspecies berkhoffii.

  Authors: Sarah A Billeter, Rick W Kasten, Lindsay F Killmaster, Edward B Breitschwerdt, Michael L Levin, Michael G Levy, Michael Y Kosoy, Bruno B Chomel

  Comparative immunology, microbiology and infectious diseases. 11/2011; 35(1):9-15.

  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.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.

• Invasion of canine erythrocytes by Bartonella vinsonii subsp. berkhoffii.

  Authors: Sarah A Billeter, Edward B Breitschwerdt, Michael G Levy

  Veterinary microbiology. 10/2011; 156(1-2):213-6.

  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,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 48h 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 48h 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.

• Expression and function of triggering receptor expressed on myeloid cells-1 (TREM-1) on canine neutrophils.

  Authors: Jingjing Li, Adam J Birkenheuer, Henry S Marr, Michael G Levy, Jeffrey A Yoder, Shila K Nordone

  Developmental and comparative immunology. 08/2011; 35(8):872-80.

  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 presentThe 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.

• Experimental infection of native north Carolina salamanders with batrachochytrium dendrobatidis.

  Authors: Sathya K Chinnadurai, David Cooper, Daniel S Dombrowski, Matthew F Poore, Michael G Levy

  Journal of wildlife diseases. 08/2009; 45(3):631-6.

  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 studiesChytridiomycosis 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.

• Evaluation of four DNA extraction methods for the detection of Tritrichomonas foetus in feline stool specimens by polymerase chain reaction.

  Authors: Stephen H Stauffer, Adam J Birkenheuer, Michael G Levy, Henry Marr, Jody L Gookin

  Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc. 09/2008; 20(5):639-41.

  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 beFeces 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.

• Detection of two Bartonella tamiae-like sequences in Amblyomma americanum (Acari: Ixodidae) using 16S-23S intergenic spacer region-specific primers.

  Authors: Sarah A Billeter, Melissa K Miller, Edward B Breitschwerdt, Michael G Levy

  Journal of medical entomology. 02/2008; 45(1):176-9.

  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 chainFour 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.

• Piscinoodinium, a fish-ectoparasitic dinoflagellate, is a member of the class dinophyceae, subclass gymnodiniphycidae: convergent evolution with Amyloodinium.

  Authors: Michael G Levy, R Wayne Litaker, Robert J Goldstein, Michael J Dykstra, Mark W Vandersea, Edward J Noga

  The Journal of parasitology. 11/2007; 93(5):1006-15.

  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 attachmentAll 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.

• Efficacy of tinidazole for treatment of cats experimentally infected with Tritrichomonas foetus.

  Authors: Jody L Gookin, Stephen H Stauffer, Maria R Coccaro, Matthew F Poore, Michael G Levy, Mark G Papich

  American journal of veterinary research. 10/2007; 68(10):1085-8.

  OBJECTIVE: To determine the efficacy of tinidazole for treatment of cats with experimentally induced Tritrichomonas foetus infection. ANIMALS: 8 specific-pathogen-free kittens. PROCEDURES: TinidazoleOBJECTIVE: 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 >or= 10 microg/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.

• Optimization of a species-specific polymerase chain reaction assay for identification of Pentatrichomonas hominis in canine fecal specimens.

  Authors: Jody L Gookin, Stephen H Stauffer, Maria R Coccaro, Miriam J Marcotte, Michael G Levy

  American journal of veterinary research. 08/2007; 68(7):783-7.

  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: DNAOBJECTIVE: 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.

• Identification of Pentatrichomonas hominis in feline fecal samples by polymerase chain reaction assay.

  Authors: Jody L Gookin, Stephen H Stauffer, Michael G Levy

  Veterinary parasitology. 05/2007; 145(1-2):11-5.

  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 presumptivelyPentatrichomonas 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.

• Detection of Rickettsia amblyommii in association with a tick bite rash.

  Authors: Sarah A Billeter, Hunter L Blanton, Susan E Little, Michael G Levy, Edward B Breitschwerdt

  Vector borne and zoonotic diseases (Larchmont, N.Y.). 02/2007; 7(4):607-10.

  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 polymeraseIn 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.

• A highly specific PCR assay for detecting the fish ectoparasite Amyloodinium ocellatum.

  Authors: Michael G Levy, Matthew F Poore, Angelo Colorni, Edward J Noga, Mark W Vandersea, R Wayne Litaker

  Diseases of aquatic organisms. 02/2007; 73(3):219-26.

  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 methodsAmyloodiniosis, 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.

• Molecular characterization of a Babesia species identified in a North American raccoon.

  Authors: Adam J Birkenheuer, Julia Whittington, Jennifer Neel, Edward Large, Anne Barger, Michael G Levy, Edward B Breitschwerdt

  Journal of wildlife diseases. 05/2006; 42(2):375-80.

  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 havePiroplasmosis 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.

• Development and evaluation of a PCR assay for the detection of Cytauxzoon felis DNA in feline blood samples.

  Authors: Adam J Birkenheuer, Henry Marr, A Rick Alleman, Michael G Levy, Edward B Breitschwerdt

  Veterinary parasitology. 05/2006; 137(1-2):144-9.

  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 diseaseCytauxzoonosis 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.

• Cytauxzoon felis infection in cats in the mid-Atlantic states: 34 cases (1998-2004).

  Authors: Adam J Birkenheuer, Jaime A Le, Amy M Valenzisi, Melissa D Tucker, Michael G Levy, Edward B Breitschwerdt

  Journal of the American Veterinary Medical Association. 03/2006; 228(4):568-71.

  OBJECTIVE: To describe the demographic and clinical characteristics of feline cytauxzoonosis in the mid-Atlantic states and compare the Cytauxzoon felis 18S rRNA gene sequences from affected catsOBJECTIVE: To describe the demographic and clinical characteristics of feline cytauxzoonosis in the mid-Atlantic 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.

• Geographic distribution of babesiosis among dogs in the United States and association with dog bites: 150 cases (2000-2003).

  Authors: Adam J Birkenheuer, Maria T Correa, Michael G Levy, Edward B Breitschwerdt

  Journal of the American Veterinary Medical Association. 10/2005; 227(6):942-7.

  OBJECTIVE: 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 wasOBJECTIVE: 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. DESIGN: Retrospective study. ANIMALS: 150 dogs. PROCEDURE: 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. RESULTS: 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. CONCLUSIONS AND CLINICAL RELEVANCE: 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.

• Surgical removal of cataracts due to Diplostomum species in Gulf sturgeon (Acipenser oxyrinchus desotoi).

  Authors: Robert S Bakal, Brian H Hickson, Brian C Gilger, Michael G Levy, James R Flowers, Lester Khoo

  Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians. 10/2005; 36(3):504-8.

  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 fishTwenty 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.

• Molecular characterization of trichomonads from feces of dogs with diarrhea.

  Authors: Jody L Gookin, Adam J Birkenheuer, Victoria St John, Michelle Spector, Michael G Levy

  The Journal of parasitology. 09/2005; 91(4):939-43.

  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 ofTrichomonads 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.

• Autochthonous visceral leishmaniasis in dogs in North America.

  Authors: Peter M Schantz, Francis J Steurer, Zandra H Duprey, Katherine P Kurpel, Stephen C Barr, Joan E Jackson, Edward B Breitschwerdt, Michael G Levy, J C Fox

  Journal of the American Veterinary Medical Association. 05/2005; 226(8):1316-22.

• Outcome of cats with diarrhea and Tritrichomonas foetus infection.

  Authors: Derek M Foster, Jody L Gookin, Matthew F Poore, Marty E Stebbins, Michael G Levy

  Journal of the American Veterinary Medical Association. 10/2004; 225(6):888-92.

  OBJECTIVE: 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.OBJECTIVE: 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. DESIGN: Prospective study. SAMPLE POPULATION: 26 cats with T. foetus-associated diarrhea at least 22 months prior to the study. PROCEDURE: 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. RESULTS: 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. CONCLUSIONS AND CLINICAL RELEVANCE: 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.