bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of the Diagnostic Sensitivity of a Commercially Available Culture Kit and a Diagnostic Culture Test Using Diamond's Media for Diagnosing Tritrichomonas Foetus in Bulls
Authors: Parker Sarah , Campbell John , Gajadhar Alvin
Published: May 2011

A number of different culture media have been described for use in the diagnosis of Tritrichomonas foetus infection in bulls, and recently, a commercial culture kit has become available. The objective of this study was to compare the sensitivity of 2 culture-based diagnostic tests for T. foetus in bulls. One test used a commercial kit for transport and culture of the samples. The other test used a thioglycollate transport medium (TFTM) for transport and a modified Diamond's medium (MDM) for culture of the samples. Twenty-one bulls infected with T. foetus were sampled repeatedly. On each sampling day, samples collected from the left and right sides of the bull were tested with one of the 2 diagnostic tests being compared. The effect of the type of diagnostic test on the outcome of the test was evaluated with a chi-square test for the calculated odds ratio. Because repeated tests from the same bull cannot be considered independent measures, unadjusted chi-square tests were adjusted for the effect of clustering by bull. Samples tested using the commercial kit were 6.95 times as likely to be positive as samples tested with a diagnostic test using MDM ( P < 0.001).
Cyanide Toxicosis in Goats after Ingestion of California Holly (Heteromeles Arbutifolia)
Authors: Tegzes John H. , Puschner Birgit , Melton Larry A.
Published: May 2011

Cyanogenic glycosides are found in many native and naturalized plants throughout North America. The glycosides themselves are not toxic, but they yield hydrogen cyanide (hydrocyanic or prussic acid) when they are hydrolyzed by β-glycosidases, either as a result of injury to the plant cells or by microbial action in the rumen. Hydrogen cyanide is readily absorbed from the gastrointestinal tract. Cyanide ion binds with iron in cytochrome oxidase, interfering with cellular respiration. The clinical effects are peracute, often resulting in death less than 1 hour after ingestion. This study describes a case that resulted in significant morbidity and mortality in a herd of goats after exposure to California holly ( Heteromeles arbutifolia).
Determination of the Median Toxic Dose of Type C Botulinum Toxin in Lactating Dairy Cows
Authors: Moeller Robert B. , Puschner Birgit , Walker Richard L. , Rocke Tonie , Galey Frank D. , Cullor James S. , Ardans Alex A.
Published: May 2011

Because of the difficulty in identifying botulinum toxin in cattle, it is hypothesized that cattle are sensitive to levels of toxin below the detection limits of current diagnostic techniques (the mouse protection bioassay and the immunostick enzyme-linked immunosorbent assay [ELISA] for type C botulinum toxin). Using an up—down method for toxicologic testing, the median toxic dose (MTD50) for cattle was determined. Four lactating Holstein cows were dosed at 0.125 or 0.25 ng/kg with Clostridium botulinum type C toxin and failed to develop clinical signs of botulism during the 7-day observation period. Three cows given 0.50 ng/kg of toxin developed clinical signs of botulism. From these results, the MTD50 was calculated at 0.388 ng/kg (3.88 mouse lethal doses/kg) using the trim-logit method. These results suggest that cattle are 12.88 times more sensitive to type C botulinum toxin than a mouse on a per kilogram weight basis. The mouse protection bioassay and the immunostick ELISA for type C botulinum toxin failed to identify the presence of the toxin in the serum, blood, and milk samples taken from all 7 animals.
Heparin in Vitro Sensitivity of the Activated Partial Thromboplastin Time in Canine Plasma Depends on Reagent
Authors: Mischke Reinhard
Published: May 2011

The in vitro heparin sensitivity of 6 different commercial activated partial thromboplastin time (APTT) reagents was investigated based on artificial plasma samples prepared by addition of sodium heparin at different activities (0—1.5 IU/ml) to pooled normal canine plasma. Statistical analysis using 2-way analysis of variance was based on APTT ratios (APTT/mean APTT control). Significant differences between the APTT ratios of different APTT reagents ( P < 0.00001) were found, which also depended on heparin activity (interaction between the factors; P < 0.00001). For example, mean APTT ratio at 0.7 IU/ml heparin varied between 1.2 and 2.5. The results of this study indicate that recommendations for the control of heparin therapy in dogs by APTT ratio should be reagent specific.
Transmission of Transmissible Mink Encephalopathy to Raccoons (Procyon Lotor) by Intracerebral Inoculation
Authors: Hamir Amir N. , Miller Janice M. , O'Rourke Katherine I. , Bartz Jason C. , Stack Mick J. , Chaplin Melanie J.
Published: May 2011

To determine the transmissibility of transmissible mink encephalopathy (TME) agent to raccoons and to provide information about clinical course, lesions, and suitability of currently used diagnostic procedures for detection of transmissible spongiform encephalopathies (TSEs) in raccoons, 4 raccoon kits were inoculated intracerebrally with a brain suspension from mink experimentally infected with TME. One uninoculated raccoon kit served as a control. All 4 animals in the TME-inoculated group showed clinical signs of neurologic disorder and were euthanized between 21 and 23 weeks postinoculation (PI). Necropsy examinations revealed no gross lesions. Spongiform encephalopathy was observed by light microscopy, and the presence of protease-resistant prion protein (PrPres) was detected by immunohistochemistry and Western blot techniques. Scrapie-associated fibrils were observed by negative-stain electron microscopy in the brains of 3 of the 4 inoculated raccoons. These findings confirm that TME is experimentally transmissible to raccoons and that diagnostic techniques currently used for TSE in livestock detect prion protein in raccoon tissue. According to previously published data, the incubation period of sheep scrapie in raccoons is 2 years, whereas chronic wasting disease (CWD) had not shown transmission after 3 years of observation. Because incubation periods for the 3 US TSEs (scrapie, TME, and CWD) in raccoons appear to be markedly different, it may be possible to use raccoons for differentiating unknown TSE agents. Retrospective genotyping of raccoons using frozen spleens showed that the raccoon PrP gene is identical to the mink gene at codons 179 and 224. Further studies, such as the incubation periods of bovine spongiform encephalopathy and other isolates of scrapie, CWD, and TME in raccoons, are needed before the model can be further characterized for differentiation of TSE agents.
Detection of Equine Herpesvirus 3 in Equine Skin Lesions by Polymerase Chain Reaction
Authors: Kleiboeker Steven B. , Chapman Rodney K.
Published: May 2011

During a recent breeding season, ulcerative, pustular skin lesions were observed on the external genitalia of 2 mares and 1 stallion within a small herd. Based on the location and description of the skin lesions plus the clinical history, equine coital exanthema, caused by equine herpesvirus 3 (EHV3), was the primary differential diagnosis. Scrapings of skin lesions from the perineum of 2 mares were submitted for diagnostic evaluation. Virus isolation was attempted by inoculation of several cell lines of equine origin, but no cytopathic agent was detected. The skin scrapings were processed for DNA extraction, and polymerase chain reaction (PCR) amplification was performed for herpesvirus DNA polymerase and DNA-packaging protein (terminase) genes using nested, degenerate primers targeted to conserved regions of the herpesvirus genome. Products of the expected sizes were generated for both assays, and subsequent nucleotide sequencing of the amplification products established that EHV3 had been detected in DNA extracted from the skin lesions. Detection of EHV3 was confirmed using an EHV3-specific PCR assay targeted to the gC gene. Using the novel EHV3 nucleotide sequence identified in this report, a sensitive and specific PCR assay targeted to the highly conserved DNA polymerase gene was developed.
Comparison of a Commercial Enzyme-Linked Immunosorbent Assay with Hemagglutination Inhibition Assay for Serodiagnosis of Swine Influenza Virus (H1N1) Infection
Authors: Skibbe Dana , Zhou En-Min , Janke Bruce H.
Published: May 2011

A commercial indirect swine influenza virus (SIV) H1N1 enzyme-linked immunosorbent assay (ELISA) was compared with the hemagglutination inhibition (HI) assay by testing 72 samples from experimentally infected pigs and 780 field samples of undefined SIV status. The HI assay was performed using SIV isolates A/Swine/IA/73 for H1N1 and A/Swine/IA/8548–1/98 for H3N2. The ELISA used an SIV isolated in 1988. The results showed that HI and ELISA detected an antibody in 11 and 6, respectively, of 72 serum samples collected from pigs experimentally infected with a 1992 SIV isolate (A/Swine/IA/40776/92). The presence of antibodies in these experimental samples was confirmed by HI tests in which all 72 samples were positive against the homologous virus, a more recent H1N1 SIV isolate (A/Swine/NVSL/01) supplied by National Veterinary Services Laboratories, Ames, Iowa, and a 1999 H1N1 isolate currently used in a commercial vaccine. On testing 780 field samples, an overall agreement of 85.5% was generated between the HI and ELISA. This study demonstrated that the ELISA is a useful serodiagnostic screening test at herd level for detecting swine antibodies against SIV. However, a new SIV isolate representing current SIV strains circulating in the field is needed to replace the older isolates used in the HI and ELISA to increase the test accuracy for serodiagnosis of SIV.
Evaluation of Pasteurella Multocida Isolated from Rabbits by Capsular Typing, Somatic Serotyping, and Restriction Endonuclease Analysis
Authors: Tayeb Amna B. El , Morishita Teresa Y. , Angrick Elisabeth J.
Published: May 2011

The goal of this study was to characterize Pasteurella multocida isolated from rabbits. Five hundred and fifty-three apparently healthy rabbits were sampled for this study. Nasal swabs were collected from each rabbit for P. multocida isolation and identification. Isolates were further characterized by capsular and somatic antigens and genomic DNA fingerprinting. Thirty-nine P. multocida isolates were recovered from 553 rabbits (7%). Capsular typing was done by depolymerization of P. multocida capsule by Staphylococcus aureus hyaluronidase and by disc diffusion with mucopolysaccharidase enzymes (heparinase III, chondroitinase AC, and hyaluronidase). Thirty-one (79%) of the isolates were capsular type A, and 8 isolates (21%) had untypable (UT) capsules. The gel-diffusion precipitin test was used to determine the somatic type of P. multocida isolates. Nineteen isolates were somatic serotype 3 (49%), 12 were serotype 1 (31%), 1 was serotype 2, 2 were serotype 5, 2 were serotype 12 with a weak reaction to antiserum raised against serotype 7 (5%), and 1 was serotype 4. Two of the isolates (5%) were UT. Restriction endonuclease analysis of the DNA of the isolates revealed 7 distinct profiles by digestion with HindIII, and 12 profiles were obtained with HpaII, whereas digestion with EcoRI did not differentiate between any of the P. multocida DNA isolates studied. The DNA restriction endonuclease enzyme HpaII was found more useful for differentiating between DNA fingerprints of P. multocida rabbit isolates. However, no correlation between capsular type, somatic serotypes, and DNA fingerprints was seen in this study.
Comparison of Manual and Automated Methods for Determining Platelet Counts in Dogs with Macrothrombocytopenia
Authors: Olsen Lisbeth H. , Kristensen Annemarie T. , Qvortrup Karen , Pedersen Henrik D.
Published: May 2011

Platelet counts were performed in 43 Cavalier King Charles Spaniels (CKCS, a breed predisposed to macrothrombocytopenia) and in 10 control dogs using 3 automated systems and 3 manual methods (erythrocyte-lysing agents + counting chamber or evaluation of blood smear). Good correlations were found between platelet counts using all methods (all P < 0.0001; R2 = 0.71–0.85). Best correlations were found between the manual methods. Significantly larger platelets were found in CKCS with platelet count ≤100,000/μl when compared with control dogs and CKCS with platelet count >100,000/μl (both P < 0.0001). All platelet counts—except when made with the 2 counting chamber methods—were underestimated a platelet counts ≤100,000/μl.
West Nile Virus Infection in Reindeer (Rangifer Tarandus)
Authors: Palmer Mitchell V. , Stoffregen William C. , Rogers Douglas G. , Hamir Amir N. , Richt Juergen A. , Pedersen Douglas D. , Waters W. Ray
Published: May 2011

West Nile virus (WNV) infection in 4 reindeer ( Rangifer tarandus) resulted in lymphohistiocytic encephalomyelitis within the medulla oblongata and cervical spinal cord. Immunohistochemistry revealed WNV antigen within neurons and among mononuclear cell infiltrates. These represent the first known cases of clinical WNV infection in Cervidae. Clinical signs and lesions were similar to those described in horses. Nucleotide sequence of a 768-bp region of the WNV E-glycoprotein gene revealed 1 nucleotide mutation, which resulted in a single amino acid substitution from a serine to a glycine (position 227 of E-glycoprotein) when compared with the prototype WNV-NY99 strain (isolated from Bronx zoo flamingo 382–99).
Effect of Temperature on the Detection of Porcine Epidemic Diarrhea Virus and Transmissible Gastroenteritis Virus in Fecal Samples by Reverse Transcription–Polymerase Chain Reaction
Authors: Jung Kwonil , Chae Chanhee
Published: May 2011

The effect of storage temperature was determined for the detection of porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) in fecal samples from experimentally and naturally infected pigs by multiplex reverse transcription–polymerase chain reaction (RT-PCR). To examine the effect of storage temperature on the ability to detect PEDV and TGEV RNA by multiplex RT-PCR, fecal samples were stored for different temperatures (4, 21, 36, and 45 C) before extracting viral RNA. The PEDV and TGEV nucleic acids in fecal samples were stable up to 3 days at 4 C. The PEDV and TGEV nucleic acids were consistently detected in fecal samples up to 60 hours at 21 C and 24 hours only at 36 and 45 C. Thereafter, the number of positive signals declined. Positive signals were detected in fecal samples stored at 4 C by 240 hours. The PEDV and TGEV nucleic acids were consistently detected in fecal samples up to 60 hours at 21 C and 24 hours only at 36 and 45 C. The results of this study suggested that storage temperature has significant effect on the detection of PEDV and TGEV nucleic acids from fecal samples by multiplex RT-PCR.
Cytology of the Normal and Abnormal Choroid Plexi in Selected Domestic Mammals, Wildlife Species, and Man
Authors: Garma-Aviña Armando
Published: May 2011

A cytologic study of the choroid plexi of animals and humans was carried out using impression smears (imprints, imp) to understand better the cellular changes that occur in the cerebrospinal fluid in the case of disease. The samples, totaling 756 imp were from 11 dogs (239 imp), 10 horses (219 imp), 1 mule (23 imp), 3 cattle (69 imp), 1 sheep (19 imp), 2 pigs (39 imp), 1 deer (20 imp), 4 monkeys (22 imp), and 7 humans (106 imp). The samples came from individuals clinically free of neurologic disease, as well as from a few abnormal cases. Six of the 7 humans had no history of neurologic disease and had a complete necropsy with brain histopathology. The seventh human case had mild neurologic signs at the time of death and only a partial necropsy with histopathological examination of the brain, in which a few small leptomeningeal lymphocytic infiltrates and polymicrogyria were found. One of the human brains without neurologic disease had arteriosclerosis. In the 40 individuals studied, several features and some unique cell types were found, for which little or no information is available. Four different morphologic cell types were consistently found in all the species studied. The first 3 types were arbitrarily named alpha (with deeply basophilic cytoplasm), beta (with neutral to weakly acidophilic cytoplasm), and gamma or vesicle-bearing cells. The third type, gamma, was a cell bearing unique inclusions (vesicles) filled with many tiny light tan to pale pink granules. The fourth type was the Kolmer cell found in very low numbers. Immature lymphocytes were found in all of 3 newborn foals, in 1 pig, and in the only stillborn calf and deer studied. The results suggest that the choroid plexi contain more than 1 epithelial cell type and that knowledge of their morphology is far from complete because other unusual cells and structures are also present in small numbers. Imprints are excellent for studying the choroid plexi, especially for tiny changes that are too subtle to be seen in hematoxylin and eosin sections.
Familial Chondrodysplasia in Holstein Calves
Authors: Agerholm Jorgen S. , Arnbjerg Jens , Andersen Ole
Published: May 2011

After the release of a report from France on the occurrence of malformed calves genetically related to a Holstein sire, a study was performed to characterize the defect. Danish breeders were encouraged to submit defective progeny of the sire for laboratory examination. Four cases were submitted, whereas a fifth case was only reported. Lesions in affected calves were analogous, with disproportionate growth retardation characterized by fascial dysplasia and shortening of the vertebral column and the abaxial skeleton. Endochondral osteogenesis was disturbed with disorganization of epiphyseal plate chondrocytes, a lesion consistent with generalized chondrodysplasia. Based on morphology, the defect was grouped as a “Dexter bulldog type”. A genetic etiology was suspected as cases occurred in a familial pattern. Genealogical evaluation revealed several common ancestors belonging to widely used breeding lines of US Holstein, but because of the extensive use of these sires, their presence in the pedigrees of affected calves might be accidental. Further studies are needed to determine the mode of inheritance.
Agreement between Three Serological Tests for Neospora Caninum in Beef Cattle
Authors: Waldner C. L. , Cunningham G. , Campbell J. R.
Published: May 2011

During 1999, serum samples were collected from beef cows on pastures in western Canada. Some of the herds had a history of confirmed abortions associated with Neospora caninum infection. All these samples were initially analyzed using a single application of 1 common commercial enzyme-linked immunosorbent assay (ELISA) for antibodies to N. caninum. From these initial results, 239 positive and 250 negative samples were randomly selected for further testing. This group of samples was retested using the 3 commercially available ELISA tests for N. caninum as per the manufacturer's recommendations. The agreement between 2 of the ELISAs was good (k = 0.76); agreement of these 2 tests with the third test was much lower (k = 0.46 and 0.60). Quantitative agreement between tests measured by intraclass correlation coefficients was also acceptable between the first 2 tests but was almost zero when the first 2 tests were compared with the third. This information is necessary to understand the differences in seroprevalence reported in different regions from laboratories using different methods.
Transmission of Sheep Scrapie to Elk (Cervus Elaphus Nelsoni) by Intracerebral Inoculation: Final Outcome of the Experiment
Authors: Hamir Amir N. , Miller Janice M. , Cutlip Randall C. , Kunkle Robert A. , Jenny Allen L. , Stack Mick J. , Chaplin Melanie J. , Richt Juergen A.
Published: May 2011

This is a final report of an experimental transmission of sheep scrapie agent by intracerebral inoculation to Rocky Mountain elk ( Cervus elaphus nelsoni). It documents results obtained in experimental ( n = 6) and control ( n = 2) elk. During the first 2 years postinoculation (PI), 3 animals died or were euthanized because of infection or injuries other than spongiform encephalopathy (SE). In years 3 and 4 PI, 3 other inoculated elk died after brief terminal neurological episodes. Necropsy of these animals revealed moderate weight loss but no other gross lesions. Microscopically, characteristic lesions of SE were seen throughout the brain and spinal cord, and the tissue was positive for proteinase K-resistant prion protein (PrPres) by immunohistochemistry (IHC) and by Western blot. Scrapie-associated fibrils (SAF) were observed by negative-stain electron microscopy in the brain of elk with neurologic signs. PrPres and SAF were not detected in the 3 inoculated elk necropsied during the first 2 years or in the 2 control animals. Retrospective analysis of the gene-encoding cervid PrP revealed a polymorphism at codon 132. The elk with SE were either homozygous (MM) or heterozygous (LM). These findings confirm that intracerebral inoculation of sheep scrapie agent results in SE with accumulations of PrPres in the central nervous system of elk. Based on morphologic and IHC findings, the experimentally induced SE cannot be distinguished from chronic wasting disease of elk with currently available diagnostic techniques.
Natural West Nile Virus Infection in a Captive Juvenile Arctic Wolf (Canis Lupus)
Authors: Lanthier Isabelle , Michel Hébert , Tremblay Donald , Josée Harel , Dallaire André D. , Girard Christiane
Published: May 2011

A case of West Nile virus (WNV) infection in a captive 4-month-old Arctic wolf ( Canis lupus) is described. The animal had vomiting, anorexia, and ataxia before death. Histopathology revealed multifocal severe renal lymphoplasmacytic vasculitis, mostly affecting small arterioles, with fibrinoid degeneration of some vessel walls. Many small foci of gliosis were detected in the cerebral cortex. West Nile virus was demonstrated in the kidneys and cerebrum by immunohistochemistry and polymerase chain reaction. The described renal changes represent a novel pathological finding of WNV infection.
Polymerase Chain Reaction–based Genetic Typing of Japanese Porcine Reproductive and Respiratory Syndrome Viruses
Authors: Yoshii Masaaki , Kaku Yoshihiro , Murakami Yosuke , Shimizu Mitsugu , Kato Kanako , Ikeda Hidetoshi
Published: May 2011

Porcine reproductive and respiratory syndrome viruses (PRRSVs) are classified into 2 distinct genotypes: the North American type and the European type. The Japanese PRRSVs were genotyped by reverse transcriptase–polymerase chain reaction using the reported primer pairs that were either reactive to both types, specific to the North American type or specific to the European type. All the PRRSV genomes from 66 tissue homogenates or sera and 55 infectious viruses were of the North American type, whereas no European-type viral genome was detected. Two PCR primers specific to the North American type showed different detection efficiency. Half of the tissue samples and 15% of the infectious viruses were not detected with one primer pair, although all of them were detected with the other primer pair. Nucleotide sequencing analysis of the forward-and reverse primer–binding sites of the nonreactive viruses indicated that all these viruses had nucleotide mismatches within the 4 bases corresponding to the 3′ end of the reverse primer. These mismatches appeared to be responsible for the nonreactivity of the former primers to these viruses.
A Comparison of Routine Culture with Polymerase Chain Reaction Technology for the Detection of Mycoplasma Species in Feline Nasal Samples
Authors: Johnson Lynelle R. , Drazenovich Nicole L. , Foley Janet E.
Published: May 2011

Nasal flush samples were collected from 20 cats and submitted for Mycoplasma culture and polymerase chain reaction (PCR). Nasal biopsy samples were also obtained from each cat and simultaneously evaluated for Mycoplasma by standard culture and PCR. Concordance of the test results was determined through calculation of the kappa statistic. In 6 cats, nasal flush samples were culture positive for Mycoplasma. PCR was positive in each culture-positive cat and also positive in 1 flush sample that was culture negative. DNA sequencing of the PCR product from the culture negative flush sample identified the organism as Mycoplasma arginini. All other flush samples that were culture negative were also PCR negative (kappa = 0.89). Nasal biopsy samples from 7 cats were culture positive for Mycoplasma, and all were PCR positive. Biopsy samples that were culture negative for Mycoplasma were also PCR negative (kappa = 1.0). Results of culture and PCR for both nasal flush and biopsy were concordant in 19 of 20 cats, and PCR was able to identify an unusual Mycoplasma species that did not grow in culture. In most cats, organisms could be detected in either nasal flush or biopsy samples. In this study, PCR provided rapid and sensitive detection of Mycoplasma species in nasal samples from cats and detected 1 organism that did not grow in culture.
Characterization of a Wild-Type Strain of Francisella Tularensis Isolated from a Cat
Authors: Inzana Thomas J. , Glindemann Gretchen E. , Snider Gerald , Gardner Susan , Crofton Lisa , Byrne Barbara , Harper Joseph
Published: May 2011

Francisella tularensis type A is the primary cause of tularemia in animals and humans in North America. The majority of research on F. tularensis has been done with the attenuated live vaccine strain (LVS), which is a type B, but very few wild-type F. tularensis strains have been characterized. A gram-negative coccobacillus that was isolated in pure culture from the lungs of a cat that died after being lost for 5 days was received for identification at the Virginia–Maryland Regional College of Veterinary Medicine Teaching hospital. The isolate (strain TI0902) was not identified (or was misidentified) by commercial identification systems; however, it was identified as F. tularensis subspecies tularensis (type A) by sequencing a portion of the 16S ribosomal RNA gene. Furthermore, repetitive extragenic palindromic sequences–polymerase chain reaction amplified a 4-kb DNA fragment from TI0902 that was characteristic of F. tularensis type A but not type B. The electrophoretic profile of the lipopolysaccharide of strain TI0902 was identical to that of the LVS by Western blotting with antiserum to LVS. The protein-enriched outer membrane of strain TI0902 contained 6–8 proteins, which were similar in molecular size to those from the LVS. Electron microscopy of negatively stained and alcian blue–stained LVS and TI0902 cells showed that both strains were coccobacillary in shape and may be encapsulated. However, after mouse challenge, the TI0902 strain was clearly more virulent than the LVS strain. Results of this study indicate that the genotype and phenotype of wild-type F. tularensis type A strain TI0902 is similar, but not identical, to that of the LVS strain. Further studies will help determine whether pathogenesis and host–pathogen interactions are also similar between the 2 strains.
Diagnosis of Taenia Saginata Cysticercosis by Immunohistochemical Test on Formalin-Fixed and Paraffin-Embedded Bovine Lesions
Authors: Ogunremi Oladele , MacDonald Garry , Geerts Stanny , Brandt Jef
Published: May 2011

A new method of diagnosing cysticercus or larval stage of the human tapeworm, Taenia saginata, also known as Cysticercus bovis, in formalin-fixed bovine tissue was developed using a monoclonal antibody to T. saginata and avidin–biotin complex immunohistochemistry. Grossly recognizable viable and degenerate cysts were identifiable after immunohistochemical staining and could be differentiated from Sarcocystis, Actinobacillus, or non-cyst, normal bovine structures. The new test should permit laboratory confirmation of suspected T. saginata cysticercus lesions.
A Comprehensive Study of Easter Lily Poisoning in Cats
Authors: Rumbeiha Wilson K. , Francis Jayaraj A. , Fitzgerald Scott D. , Nair Muraleedharan G. , Holan Kate , Bugyei Kwasi A. , Simmons Heather
Published: May 2011

This study was conducted with 3 objectives in mind: first, to identify the toxic fraction (aqueous or organic) in leaves and flowers; second, to identify diagnostic marker(s) of toxicosis in cats; and, third, to evaluate the morphologic effects of intoxication. The study was conducted in 2 phases. Phase 1 was to identify which extract, organic or aqueous, was nephrotoxic and also to determine the appropriate dose for use in the phase 2 studies. Results indicated that only the aqueous extracts of leaves and flowers were nephrotoxic and pancreotoxic. To identify the proximate toxic compound, cats in the phase 2 study were orally exposed to subfractions of the aqueous flower extract, 1 subfraction per cat. Results confirmed vomiting, depression, polyuria, polydipsia, azotemia, glucosuria, proteinuria, and isosthenuria as toxic effects of the Easter lily plant. Another significant finding in serum was elevated creatinine kinase. Significant histologic kidney changes included acute necrosis of proximal convoluted tubules and degeneration of pancreatic acinar cells. Renal ultrastructural changes included swollen mitochondria, megamitochondria, edema, and lipidosis. Subfraction IIa3 of the aqueous floral extract contained most of the toxic compound(s). These studies reproduced the clinical disease, identified the most toxic fraction of the Easter lily, and helped characterize the clinical pathology, histopathology, and ultrastructural pathology associated with the disease.
Expression of the KIT Protein (CD117) in Primary Cutaneous Mast Cell Tumors of the Dog
Authors: Preziosi Rosario , Morini Maria , Sarli Giuseppe
Published: May 2011

Thirty-one canine cutaneous masses, diagnosed as mast cell tumors (MCT) by histopathologic analysis, were used to evaluate the immunohistochemical pattern of expression of KIT protein (CD117), a type III tyrosine kinase protein involved in mast cell growth and differentiation. Lesions were graded as I (well differentiated), II (intermediate differentiation), or III (poorly differentiated) according to the following morphologic features: invasiveness, cellularity and cellular morphology, mitotic index, and stromal reaction. Immunohistochemical KIT expression was compared with histologic grade and some histomorphologic features (cell differentiation and nuclear grade) evaluated separately. A possible predictive role of biologic behavior in MCTs for KIT expression was also investigated. Immunohistochemical analysis revealed three different patterns of KIT expression: a cytoplasmic diffuse pattern, a membranous pattern with immunostaining located on the cell surface, and a cytoplasmic perinuclear pattern, where KIT expression was detected in the cytoplasm of the neoplastic mast cells, close to the nucleus. Statistical analysis showed a close relationship between different KIT immunohistochemical patterns and histologic grade ( P < 0.00000), cell differentiation ( P < 0.00000), and nuclear grade ( P < 0.0024). According to Kaplan-Meier–estimated survival curves compared by survival analysis, KIT expression was significantly associated with survival time ( P = 0.037) but not cancer-free interval ( P = 0.50). Similar to other well-known histomorphological features, KIT expression is a useful parameter of malignancy in cutaneous MCTs. KIT expression also predicted the biological behavior of the tumors in this study.
Diagnosis of a Mixed Mycoplasma Infection Associated with a Severe Outbreak of Bovine Pinkeye in Young Calves
Authors: Levisohn Sharon , Garazi Shlomo , Gerchman Irinna , Brenner Jacob
Published: May 2011

Mycoplasma bovoculi and Mycoplasma bovis were both isolated from conjunctival swabs taken from young calves showing symptoms consistent with infectious bovine keratoconjunctivitis (pinkeye). No Moraxella spp. or other nonmycoplasma bacteria were isolated in association with this severe clinical outbreak. Based on laboratory tests and clinical observations, the first phase of the disease was likely pneumonic in nature, possibly caused by bovine respiratory syncytial virus and M. bovis. In the subsequent phase of the disease course, infection with both M. bovoculi and M. bovis resulted in ocular disease. A combination of microbiological, serological, and molecular diagnostic methods was used to elucidate the etiology of the outbreak.
Design, Validation, and Absolute Sensitivity of a Novel Test for the Molecular Detection of Avian Pneumovirus
Authors: Cecchinato Mattia , Catelli Elena , Savage Carol E. , Jones Richard C. , Naylor Clive J.
Published: May 2011

This study describes attempts to increase and measure sensitivity of molecular tests to detect avian pneumovirus (APV). Polymerase chain reaction (PCR) diagnostic tests were designed for the detection of nucleic acid from an A-type APV genome. The objective was selection of PCR oligonucleotide combinations, which would provide the greatest test sensitivity and thereby enable optimal detection when used for later testing of field materials. Relative and absolute test sensitivities could be determined because of laboratory access to known quantities of purified full-length DNA copies of APV genome derived from the same A-type virus. Four new nested PCR tests were designed in the fusion (F) protein (2 tests), small hydrophobic (SH) protein (1 test), and nucleocapsid (N) protein (1 test) genes and compared with an established test in the attachment (G) protein gene. Known amounts of full-length APV genome were serially diluted 10-fold, and these dilutions were used as templates for the different tests. Sensitivities were found to differ between the tests, the most sensitive being the established G test, which proved able to detect 6,000 copies of the G gene. The G test contained predominantly pyrimidine residues at its 3' termini, and because of this, oligonucleotides for the most sensitive F test were modified to incorporate the same residue types at their 3' termini. This was found to increase sensitivity, so that after full 3' pyrimidine substitutions, the F test became able to detect 600 copies of the F gene.
Copper-Associated Hepatopathy in a Mexican Fruit Bat (Artibeus Jamaicensis) and Establishment of a Reference Range for Hepatic Copper in Bats
Authors: Hoenerhoff M. , Williams K.
Published: May 2011

Copper toxicity has been described in numerous domestic species. The characteristic lesions include hemoglobinuric nephrosis and piecemeal hepatic necrosis with bile ductular hyperplasia and portal fibrosis. Certain species, such as sheep, are prone to toxicity when exposed to copper in feed, whereas an inherent genetic defect of copper storage is present in some breeds of dogs (Bedlington Terriers, West Highland White Terriers, Doberman Pinschers). In nondomestic species, reference ranges have not been established for copper in internal organs, so the establishment of copper toxicity as a diagnosis is difficult. A case of copper toxicity in a captive Mexican fruit bat is presented. Hepatic copper levels in 16 additional bats, of at least 3 different species, were measured. To the authors' knowledge, this is the first reported case of copper toxicity in a chiropteran.
Genetic and Clinical Analyses of Bovine Viral Diarrhea Virus Isolates from Dairy Operations in the United States of America
Authors: Tajima Motoshi , Dubovi Edward J.
Published: May 2011

To assess the prevalence of bovine viral diarrhea virus (BVDV) on the basis of the genotype and clinical signs, isolates from 16 dairy herds (bulk milk samples) and 37 BVDV-infected cattle were examined. Isolates for this study were selected from submissions that contained an adequate clinical history. A part of the E2 gene of BVDV from these isolates was amplified by reverse transcription-polymerase chain reaction. From the nucleotide sequence of the amplified products, phylogenetic analyses were performed and genotypes or subgenotypes were identified. Forty percent of the selected field isolates were BVDV-2, and 60% were BVDV-1. Eighty-one percent of BVDV-1 isolates were determined to be the BVDV-1b subgenotype. BVDV-1b and BVDV-2 formed more closely related clusters in each group than did the BVDV-1a isolates. There was no obvious association of any genotype or subgenotype with geographical localization or clinical manifestations. A higher prevalence of BVDV-2 infection was found in the United States than in other countries. BVDV-1a has been thought of as a prototype of BVDV; however, there were fewer isolations of BVDV-1a than of other subgenotypes of BVDV Phylogenetic analyses of BVDV isolates using the E2 region of the genome generated results similar to those of studies done in the United States using the 5' untranslated region.
Vulvar Lipoleiomyoma in a Dog
Authors: Radi Zaher A.
Published: May 2011

A vulvar neoplasm from a 5-year-old female Siberian Husky dog was removed surgically and examined histologically. Macroscopically, the neoplasm was firm, white, and measured 6 × 4 × 3 cm. Microscopically, the neoplasm was expansile, nonencapsulated, and composed of lobules of mature adipocytes admixed with streams and bundles of well-differentiated smooth muscle cells. Immunohistochemically, the neoplastic cells had strong diffuse cytoplasmic immunoreactivity for α-smooth muscle actin and desmin, and no immunoreactivity for cytokeratin or vimentin. On the basis of gross, histopathologic, and immunohistochemical findings, a diagnosis of lipoleiomyoma was made. This is, to the author's knowledge, the first report of canine vulvar lipoleiomyoma.
Persistent Metanephric Ducts in a Geriatric White Tiger
Authors: de Bruin Alain , Wack Ray , Weisbrode Steven E.
Published: May 2011

An 18-year-old male intact white Tiger ( Panthera tigris) was euthanized after a clinical diagnosis of severe renal failure. Postmortem macroscopic examination of the kidneys revealed unilateral hydronephrosis with renal calculi and bilateral cortical and medullary fibrosis and papillary coagulation necrosis. Interestingly, multiple large persistent metanephric ducts were found at the corticomedullary junctions accompanied by marked interstitial fibrosis, tubular atrophy, and lymphoplasmacytic interstitial nephritis. To our knowledge, this is the first reported case of persistent metanephric ducts in cats.
Simultaneous Detection of North American and European Porcine Reproductive and Respiratory Syndrome Virus Using Real-Time Quantitative Reverse Transcriptase–PCR
Authors: Kleiboeker Steven B. , Schommer Susan K. , Lee Sang-Myeong , Watkins Sandy , Chittick Wayne , Polson Dale
Published: May 2011

Porcine reproductive and respiratory syndrome (PRRS) is 1 of the most economically important diseases of swine. Detection of the etiologic agent, PRRS virus (PRRSV), represents a diagnostic challenge due to the heterogeneity of field isolates as well as the propensity for swine to develop persistent infection in which virus is difficult to detect. Recently European (EU) lineage PRRSV isolates, which are genetically divergent from North American (NA) isolates, have been introduced into NA swine further complicating efforts to diagnose this disease. In this study, real-time ( TaqMan) reverse transcriptase (RT)–PCR assays were developed for multiplex detection, differentiation, and quantification of NA and EU PRRSV field isolates. Oligonucleotide primers and dual-labeled probes were selected from conserved regions of open-reading frame 7 and the 3'-untranslated region. The real-time RT-PCR assays described for the NA or EU genotype of PRRSV detected viral RNA from 83/83 strains (74 NA; 9 EU) previously isolated by cell culture between 1992 and 2003. The analytical sensitivity of both assays was consistently found to be less than a single TCID50, which corresponded to 5–10 RNA molecules, and was not significantly reduced when the reactions were performed in a multiplex format. When performing multiplex reactions, sensitive detection was possible even when 1 viral RNA concentration was up to 5,000-fold higher than the second. The diagnostic sensitivity and specificity of the multiplex reaction was found to be at a minimum equivalent to that of both nested RT-PCR and virus isolation.
Features of Porcine Circovirus-2 Disease: Correlations between Lesions, Amount and Distribution of Virus, and Clinical Outcome
Authors: Krakowka Steven , Ellis John , McNeilly Francis , Waldner Cheryl , Allan Gordon
Published: May 2011

Tissue sets from 36 snatch-farrowed colostrum-deprived (SF/CD) and 71 Caesarian-derived gnotobiotic swine infected with porcine circovirus type 2 (PCV-2) as neonates were examined and scored for the types and tissue distribution of histologic lesions associated with this viral infection. The occurrence and severity of these lesions were correlated with qualitative and quantitative determinations of viral burden in tissues by immunohistochemistry (IHC) and tissue titrations for infectious virus, respectively. These measures were, in turn, related to 1 of 3 categories of clinical disease expressed in PCV-2–infected swine as subclinical infection, preclinical postweaning multisystemic wasting syndrome (PMWS), and clinically evident PMWS, respectively. Statistically significant ( P < 0.05 to 0.001) associations between both measures of viral burden, the severity of histologic lesions and the stage of disease were obtained. Discrimination between and among categories of disease was best accomplished by a combination of IHC and histopathology. The results of this study confirm that viral burden in PCV-2–infected tissues, specifically lymphoid tissues and liver, directly correlate with severity of clinical disease expression in PCV-2 infected swine.
Psittacine Plasma Concentrations of Elements: Daily Fluctuations and Clinical Implications
Authors: Rosenthal Karen L. , Johnston Matthew S. , Shofer Frances S. , Poppenga Robert H.
Published: May 2011

During the past 2 decades, the potential for excessive exposure of pet birds to zinc has become a concern for many pet bird owners. Ideally, avian zinc toxicosis is diagnosed on the basis of history of exposure to zinc, radiographic evidence of ingested metal, occurrence of melena, detection of an elevated plasma zinc concentration, and response to treatment. However, most pet birds suspected of having zinc toxicosis present with vague signs and lack of radiographic evidence; therefore, the diagnosis relies on the presence of an elevated plasma zinc concentration. A question was, is there a significant diurnal variation in the zinc concentration in psittacine birds and could this be clinically relevant? Because studies in other species have shown that zinc is not the only element that shows a diurnal variation, the authors examined 13 other plasma elements including arsenic, cadmium, calcium, cobalt, copper, iodine, iron, magnesium, manganese, molybdenum, potassium, selenium, and total phosphorus. Fifteen adult psittacine birds housed in the same aviary were used in this study. Three blood samples, separated by 4 hours, were taken from the right jugular vein in each bird. All elements were measured in plasma. Zinc, copper, and molybdenum revealed diurnal fluctuations. The results of this study suggest that interpretation of clinical samples may be more complicated than previously believed. Furthermore, on the basis of the results of this study, it is possible that some avian reference ranges may need to be reexamined.
Retroperitoneal Perirenal Pseudocyst in a Massese Breed Ram
Authors: Mutinelli Franco , Vascellari Marta , Schiavon Eliana
Published: May 2011

The macroscopic and microscopic features of a retroperitoneal perirenal pseudocyst in a 12-month-old ram without impairment of renal function are described. In humans and animals, uriniferous pseudocysts may be of traumatic origin, resulting from rupture of kidney, renal pelvis, or ureter, or congenital. Lymphatic pseudocysts may develop secondary to inflammatory obstruction of the hilar lymphatics after perinephritis or renal transplantation. In this case, histologic characteristics of the pseudocyst wall were suggestive of development from the parietal peritoneal layer encapsulating the kidney. This is the first case of retroperitoneal perirenal pseudocyst in a sheep.
Frequency, Body Distribution, and Population Size of Malassezia Species in Healthy Dogs and in Dogs with Localized Cutaneous Lesions
Authors: Cafarchia Claudia , Gallo Sabrina , Romito Diana , Capelli Gioia , Chermette René , Guillot Jacques , Otranto Domenico
Published: May 2011

Malassezia species are commensal organisms of human and animal skin that occasionally act as opportunistic pathogens. The lipid-dependent species are associated with human skin disorders, whereas the non–lipid-dependent species ( Malassezia pachydermatis) is considered as an opportunistic secondary pathogen affecting the canine skin surface and ear canal. This study evaluated the relationship between Malassezia yeasts, their population size, and the occurrence of skin lesions from healthy and skin-diseased dogs. The efficiency of cytological examination and fungal culture for Malassezia detection was also evaluated. From March 2002 to July 2003, 33 healthy dogs and 54 dogs with pruritic localized skin diseases were examined; skin swabs (1218) were collected from 7 anatomical sites for culture and cytological examination. Malassezia prevalence according to anatomical site and the agreement between cytological results and fungal cultures were statistically analyzed. Differences in mean colony forming unit counts between positive healthy and diseased dogs were evaluated using the Bonferroni test for post hoc pair-wise comparisons. In healthy dogs, Malassezia yeasts were most frequently isolated in the perianal and perioral areas. The frequency of isolation and population size of Malassezia species were higher in dogs with localized dermatitis, especially in affected areas, indicating a role for Malassezia in the occurrence of skin lesions. Malassezia pachydermatis was the species most commonly cultured from the skin and external ear canal of healthy and diseased dogs; isolation of lipid-dependent yeasts from healthy dogs was less frequent. Using fungal culture as the gold standard, cytological examination showed good relative specificity (95%) but very low relative sensitivity (30%).
Adenocarcinoma of Apocrine Sweat Glands in a Mouflon (Ovis Musimon)
Authors: Morandi Federico , Benazzi Cinzia , Simoni Paolo
Published: May 2011

A free-living mouflon ( Ovis musimon) was presented with a mass on the left shoulder. At necropsy, multifocal, slightly protruding whitish spots were noted on the kidneys, and several lymph nodes were abnormal. Histologically, the mass was composed of epithelial cells arranged in tubular and tubulopapillary structures. The cytoplasm of the epithelial cells had numerous periodic acid–Schiff-positive and diastase-resistant granules. Ultrastructurally, the cytoplasm of the neoplastic cells contained numerous pleomorphic secretory granules and microvilli, which partially covered the luminal surface of the tumor cells. Metastatic foci were present in pres-capular and mediastinal lymph nodes and kidneys. On the basis of histological and ultrastructural findings, this tumor was diagnosed as a tubulopapillary adenocarcinoma, arising from apocrine sweat glands of the skin.