bluuebunny/crossref_metadata_2025 huggingface dataset:
Ovarian mucinous cystadenoma in a gray brocket deer (Mazama gouazoupira)
Authors: Monteiro Lidianne N. , Salgado Breno S. , Grandi Fabrizio , Fernandes Thaís R. , Miranda Bruna S. , Teixeira Carlos R. , Rocha Rafael M. , Rocha Noeme S.
Published: May 2011

An ovarian mucinous cystadenoma is described in a gray brocket deer ( Mazama gouazoupira). The tumor was histologically characterized by the presence of cysts and proliferation of papillae, both lined by single- or multi-layered pleomorphic epithelial cells that contained alcian blue–positive mucins.
Use of a multiplex enzyme-linked immunosorbent assay to detect a subpopulation of Mycobacterium bovis–infected animals deemed negative or inconclusive by the single intradermal comparative tuberculin skin test
Authors: Whelan Clare , Shuralev Eduard , Kwok Hang Fai , Kenny Kevin , Duignan Anthony , Good Margaret , Davis William C. , Clarke John
Published: May 2011

Although the single intradermal comparative tuberculin skin test (SICTT) remains the most effective assay for detecting cattle infected with Mycobacterium bovis, not all infected animals are detected with the SICTT. This has made it difficult to control bovine tuberculosis using a single assay. Use of the gamma interferon assay in conjunction with the SICTT has improved the level of detection but some infected animals still go undetected. This could be in part attributable to both assays being reliant on a cell-mediated immune response. The present study was undertaken to determine if a multiplex assay can improve the level of detection of infected animals when used in combination with the SICTT. The Enferplex TB assay is a multi-antigen ELISA designed for the detection of antibody in animals at different stages of infection and disease. Sixty cattle that were confirmed by histopathology and/or culture to be infected with M. bovis and that were SICTT negative (43.3%) or difficult to evaluate (56.7% inconclusive) were used in the study. Fifty-three (88.3%) of the animals were positive in multiplex ELISA. The results show that the level of detection of M. bovis–infected animals can be improved by the combined use of the SICTT and the multiplex ELISA.
Preliminary validation of assays to measure parameters of calcium metabolism in captive Asian and African elephants in western Europe
Authors: van Sonsbeek Gerda R. , van der Kolk Johannes H. , van Leeuwen Johannes P.T.M. , Schaftenaar Willem
Published: May 2011

Hypocalcemia is a well known cause of dystocia in animals, including elephants in captivity. In order to study calcium metabolism in elephants, it is of utmost importance to use properly validated assays, as these might be prone to specific matrix effects in elephant blood. The aim of the current study was to conduct preliminary work for validation of various parameters involved in calcium metabolism in both blood and urine of captive elephants. Basal values of these parameters were compared between Asian elephants ( Elephas maximus) and African elephants ( Loxodonta africana). Preliminary testing of total calcium, inorganic phosphorus, and creatinine appeared valid for use in plasma and creatinine in urine in both species. Furthermore, measurements of bone alkaline phosphatase and N-terminal telopeptide of type I collagen appeared valid for use in Asian elephants. Mean heparinized plasma ionized calcium concentration and pH were not significantly affected by 3 cycles of freezing and thawing. Storage at 4°C, room temperature, and 37°C for 6, 12, and 24 hr did not alter the heparinized plasma ionized calcium concentration in Asian elephants. The following linear regression equation using pH (range: 6.858–7.887) and ionized calcium concentration in heparinized plasma was utilized: iCa7.4 (mmol/l) = –2.1075 + 0.3130·pHactual + 0.8296·iCaactual (mmol/l). Mean basal values for pH and plasma in Asian elephant whole blood were 7.40 ± 0.048 and 7.49 ± 0.077, respectively. The urinary specific gravity and creatinine concentrations in both Asian and African elephants were significantly correlated and both were significantly lower in Asian elephants.
Evaluation of the Mythic 18 hematology analyzer for use with canine, feline, and equine samples
Authors: Wassmuth Andrea K. , Riond Barbara , Hofmann-Lehmann Regina , Lutz Hans
Published: May 2011

The Mythic 18 is a fully automated hematology bench-top analyzer using impedance technology for a complete blood cell count (CBC) and a 3-part white blood cell count (WBC) differential. The purpose of the current study was to evaluate the Mythic for assessment of agreement, precision, linearity, carry-over, stability, and usability under practice conditions. Ethylenediamine tetra-acetic acid–blood samples from 122 dogs, 140 cats, and 123 horses were analyzed with the Mythic and reference methods (Sysmex XT-2000iV, manual hematocrit, and microscopic WBC differentiation). Pearson’s coefficient of correlation, Passing–Bablok regression analysis, and Bland–Altman difference plots were performed to determine agreement. For precision, standard deviation and coefficients of variation were calculated. Linearity was determined according to Emancipator–Kroll. Red blood cell parameters showed excellent correlation and small biases, except for red cell distribution width and mean corpuscular hemoglobin concentration. Total WBC correlated excellently in canine and equine and very well in feline samples. In 23 feline specimens with platelet aggregates, the Mythic overestimated WBC. In all 3 species, absolute granulocyte counts correlated excellently. Equine lymphocyte counts showed good correlation whereas canine and feline lymphocyte counts correlated poorly. Feline platelets showed good correlation with a negative bias. The instrument showed good to excellent precision. The whole 3-part differential was found to be accurate in horses. In dogs and cats, absolute granulocyte counts were reliable. As with all impedance-based hematological instruments, evaluation of a blood smear is absolutely indicated to check for the presence of platelet aggregates, to verify WBC differentiation, and to identify possible abnormalities.
Measurement of serum anti-Müllerian hormone concentration in female dogs and cats before and after ovariohysterectomy
Authors: Place Ned J. , Hansen Betty S. , Cheraskin Jeri-Lyn , Cudney Sarah E. , Flanders James A. , Newmark Andrew D. , Barry Bridget , Scarlett Janet M.
Published: May 2011

Anti-Müllerian hormone (AMH), or Müllerian inhibitory substance, is a hormone that is best known for its production by fetal testes in mammals and as the inhibitor of Müllerian (paramesonephric) duct development in males. However, following the development of the Müllerian ducts into the oviduct, uterus, and upper vagina in female mammals, the ovaries produce AMH, which can be found in measureable amounts within the peripheral circulation, especially in adults. The ovaries appear to be the sole source of AMH in the circulation; therefore, it may be a useful marker in clinically relevant situations when an assessment of the presence or absence of ovaries or ovarian remnants in dogs and cats is important. To that end, a commercially available, human-based assay was evaluated for the measurement of AMH in dogs and cats. A preliminary assessment involved a single test on a set of serum samples from dogs that were submitted to a diagnostic endocrinology laboratory for other tests. Favorable preliminary results led to a more formal assessment of the assay using serum samples from dogs and cats with the presence or absence of the ovaries known by surgical confirmation. Overall, a single measurement of serum AMH concentration was highly effective at distinguishing ovariohysterectomized from intact adult animals. In addition, the assay also accurately identified several cases of ovarian remnant syndrome.
Canine parvovirus-2b–associated erythema multiforme in a litter of English Setter dogs
Authors: Woldemeskel Moges , Liggett Alan , Ilha Marcia , Saliki Jeremiah T. , Johnson Leslie P.
Published: May 2011

Erythema multiforme (EM) was diagnosed in a litter of English Setter puppies. The puppies developed erythematous cutaneous lesions at the age of 2 weeks. Microscopically, there was individual keratinocyte apoptosis associated with lymphocyte exocytosis in all layers of the epidermis. Intranuclear viral inclusions were seen in multiple tissues and organs. Tissues from the tongue, lymph node, spleen, skin, and small intestine were positive for Canine parvovirus-2 (CPV-2) and negative for Canine distemper virus (CDV) and Canid herpesvirus 1 by fluorescent antibody test. Negative-staining electron microscopy detected parvovirus particles in the intestinal contents. The skin and small intestine were positive for CPV-2b and negative for CDV by polymerase chain reaction. The mucocutaneous junctions and small intestines stained positive for CPV by immunohistochemistry. The present report documents CPV-2b–associated EM in a litter of English Setters and substantiates the single previous report associating EM with CPV-2. The finding suggests that CPV should be considered as a possible cause of EM in dogs.
The use of meat juice or blood serum for the diagnosis of Salmonella infection in pigs and its possible implications on Salmonella control programs
Authors: Vico Juan P. , Mainar-Jaime Raúl C.
Published: May 2011

Serology is the method of choice for country-scale Salmonella control programs in pigs and can be carried out both on blood serum or meat juice. However, the diagnostic performance of enzyme-linked immunosorbent assays (ELISAs) on these sample matrices has not been sufficiently compared. The agreement between the serum ELISA and meat juice ELISA on samples taken from commercial farms was assessed in 2 pig populations (adult sows and finishers). Results of optical density percentage (OD%) from the serum ELISA were consistently higher than those from the meat juice ELISA (38.5 vs. 27.9; P < 0.001), and the mean difference between them was significantly different from zero ( P < 0.0001). The overall correlation coefficient between serum ELISA and meat juice ELISA results was low ( r = 0.53). These results indicated an important disagreement between ELISA performed on serum and meat juice matrices and suggested that before implementing a control program to reduce the prevalence of Salmonella in swine the choice of matrix on which to perform the ELISA should be carefully considered.
Polymerase chain reaction–based identification of clinically relevant Pasteurellaceae isolated from cats and dogs in Poland
Authors: Król Jarosław , Bania Jacek , Florek Magdalena , Pliszczak-Król Aleksandra , Staroniewicz Zdzisław
Published: May 2011

A set of polymerase chain reaction (PCR) assays for identification of the most important Pasteurellaceae species encountered in cats and dogs were developed. Primers for Pasteurella multocida were designed to detect a fragment of the kmt, a gene encoding the outer-membrane protein. Primers specific to Pasteurella canis, Pasteurella dagmatis, and Pasteurella stomatis were based on the manganese-dependent superoxide dismutase gene ( sodA) and those specific to [ Haemophilus] haemoglobinophilus on species-specific sequences of the 16S ribosomal RNA gene. All the primers were tested on respective reference and control strains and applied to the identification of 47 canine and feline field isolates of Pasteurellaceae. The PCR assays were shown to be species specific, providing a valuable supplement to phenotypic identification of species within this group of bacteria.
Polymerase chain reaction–restriction fragment length polymorphism analysis of the genes involved in the biosynthesis of the lipopolysaccharide of Pasteurella multocida
Authors: Tsai Yu-Chen , Shien Jui-Hung , Wu Jin-Ru , Shieh Happy K. , Chang Poa-Chun
Published: May 2011

The lipopolysaccharide, also known as the somatic antigen or O-antigen, is an important virulence factor of Pasteurella multocida. In the current study, the genes involved in the biosynthesis of the outer core region of the lipopolysaccharide, which were obtained from somatic type reference strains and field strains of P. multocida, were subjected to polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis. The PCR-RFLP analysis classified 11 out of the 16 serotypes into 5 PCR-RFLP types (I–V). Types I and V contain strains belong to serotypes 1 and 13, respectively. The rest of the PCR-RFLP types contain strains belong to certain groups of serotypes. Typing of 38 field strains from poultry using PCR-RFLP analysis and the gel diffusion precipitation test showed consistent results. These results indicate that the PCR-RFLP analysis can be a useful tool for rapid somatic typing of some strains of P. multocida.
Optimization of Betanodavirus culture and enumeration in striped snakehead fish cells
Authors: Hick Paul , Tweedie Alison , Whittington Richard J.
Published: May 2011

An optimized culture method for detection of infection of fish with the Red spotted grouper nervous necrosis virus (RGNNV) genotype of betanodavirus in striped snakehead (SSN-1, Channa striatus) cells is described. Inoculation of fish tissue homogenates at the same time or within 4 hr of seeding the SSN-1 cells was as sensitive as the method recommended by the World Organization for Animal Health, where homogenates were adsorbed onto an established cell monolayer. Such modification halved the time required and the costs of consumables, and reduced the potential for error when processing large numbers of samples. Positive culture results were obtained from 88.3% of 392 fish tissue homogenates in which RGNNV was detected using a quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay; 99.7% of 943 tissue homogenates, which were qRT-PCR negative, were cell culture negative. Cytopathic effect (CPE) was characterized by large intracytoplasmic vacuoles in 0.1–60% of cells. Detachment of affected cells from the culture surface resulting in progressive disruption of the monolayer occurred in 46.4% of primary cultures and 96.0% of subcultures of positive samples. Identification of CPE that did not disrupt the cell monolayer increased estimates of the 50% tissue culture infective dose (TCID50) by 1.07–2.79 logs (95% confidence interval). The predicted mean TCID50/ml was 3.3 logs higher when cells were inoculated less than 36 hr after subculture at less than 80% confluence compared to cells inoculated at greater than 80% confluence and more than 36 hr after subculture ( P < 0.05).
Expression of a secreted version of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus
Authors: Zoth Silvina Chimeno , Gómez Evangelina , Carballeda Juan Manuel , Carrillo Elisa , Berinstein Analía
Published: May 2011

The hemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus (NDV) constitutes, together with the fusion glycoprotein, the main surface antigen of this avian pathogen, which causes a highly contagious disease, relevant economically worldwide. The purpose of this work was to obtain the HN glycoprotein as a soluble antigen in culture supernatants of recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells and to evaluate its application to the development of a recombinant enzyme-linked immunosorbent assay (rELISA) for the analysis of chicken sera. A transfer vector for baculovirus containing the sequence of a melittin signal peptide was constructed and the sequence coding for HN protein without its own signal peptide was cloned. The recombinant protein was secreted and recovered easily from the culture medium of Sf9-infected cells. The recombinant protein was evaluated as antigen for ELISA coating the plates with the recovered HN using 79 positive and 142 negative samples. The Cohen kappa value resulted 0.91, indicating excellent agreement between the rELISA and the hemagglutinin inhibition tests. The rELISA was also compared with a commercial ELISA, finding high levels of agreement between both assays. The present results show that the cloning strategy developed yielded the HN protein free in the cell culture supernatant and that the recombinant protein retained its reactivity with anti-NDV HN antibodies in chicken sera.
In vitro antimicrobial inhibition of Mycoplasma bovis isolates submitted to the Pennsylvania Animal Diagnostic Laboratory using flow cytometry and a broth microdilution method
Authors: Soehnlen Marty K. , Kunze M. Elaine , Karunathilake K. Eranda , Henwood Brittnee M. , Kariyawasam Subhashinie , Wolfgang David R. , Jayarao Bhushan M.
Published: May 2011

Mycoplasma bovis is an important pathogen of cattle, causing mastitis, pneumonia, conjunctivitis, otitis, and arthritis. Currently there are only a few reports of sensitivity levels for M. bovis isolates from the United States. Mycoplasma bovis isolates submitted to the Pennsylvania Animal Diagnostic Laboratory between December 2007 and December 2008 (n = 192) were tested for antimicrobial susceptibility to enrofloxacin, erythromycin, florfenicol, spectinomycin, ceftiofur, tetracycline, and oxytetracycline using a broth microdilution method. The most effective antimicrobials against M. bovis determined by using the broth microdilution method were florfenicol, enrofloxacin, and tetracycline with minimum inhibitory concentration (MIC) ranges of 2–32 µg/ml, 0.1–3.2 µg/ml, and 0.05 to >12.8 µg/ml, respectively. Spectinomycin, oxytetracycline, and tetracycline showed a wide-ranging level of efficacy in isolate inhibition with broth microdilution with MIC ranges of 4 to >256 µg/ml, 0.05 to >12.8 µg/ml, and 0.05 to >12.8 µg/ml, respectively. A significant difference in the susceptibility levels between quarter milk and lung isolates was found for spectinomycin. When MIC values of a subset of the M. bovis isolates (n = 12) were tested using a flow cytometric technique, the MIC ranges of enrofloxacin, spectinomycin, ceftiofur, erythromycin, tetracycline, oxytetracycline, and florfenicol ranges were 0.1–0.4 µg/ml, 4 to >256 µg/ml, >125 µg/ml, >3.2 µg/ml, <0.025 to >6.4 µg/ml, 0.8 to >12.8 µg/ml, and <2–4 µg/ml, respectively. Flow cytometry offers potential in clinical applications due to high-throughput capability, quick turnaround time, and the objective nature of interpreting results.
Attaching and Effacing Escherichia coli Infections in Calves, Pigs, Lambs, and Dogs
Authors: Janke Bruce H. , Francis David H. , Collins James E. , Libal Melissa C. , Zeman David H. , Johnson Darrell D.
Published: May 2011

Attaching and effacing Escherichia coli (AEEC) adhere to mucosal epithelium in both small and large intestine and induce a distinctive lesion characterized by an irregular scalloped appearance of the epithelial layer. Infection with attaching and effacing E. coli was detected in 14 calves, 7 pigs, 2 lambs, and 3 dogs. Affected animals were from farms and kennels in South Dakota, Minnesota, Iowa, Nebraska, and Wisconsin. Ages of affected animals were calves, 2 days to 4 months; pigs, 1–6 weeks; lambs, 1 week; and dogs, 7–8 weeks. Clinical signs included diarrhea in all animals, but other nonenteric disease problems were present in some animals. Concurrent infection with other enteropathogens was detected in 9 calves and 5 pigs. Infection with AEEC appeared to be the sole cause of illness and death in some animals. There was evidence of intestinal hemorrhage in 5 of the calves and in all 3 dogs. Attaching and effacing lesions varied from small scattered foci to widespread involvement of large areas of intestinal mucosa. Verotoxin was produced by E. coli strains isolated from 9 calves, but not by strains from pigs, lambs, or dogs.
Actinobacillus (Haemophilus) Pleuropneumoniae: Use of Coagglutination and Complement Fixation to Determine the Relationship between Presence of Organism and Antibody Titer in Slaughterhouse Pigs
Authors: Hoffman Lorraine J.
Published: May 2011

The conventional culture method was compared to coagglutination for detection of Actinobacillus (Haemophilus) pleuropneumoniae in 425 sets of pig lungs. Sera from the same animals were evaluated for antibodies to A. pleuropneumoniae by the complement fixation (CF) test. All samples were collected at 2 packing plants in Iowa. In 2 nonvaccinated herds with no history of respiratory disease, the difference between standard culture results and coagglutination was highly significant ( P < 0.001). None of the 57 pigs in this group were positive for A. pleuropneumoniae by conventional culture, but 7 were positive by the coagglutination test. There were 15 animals with CF titers between 1:8 and 1:32. Animals from 6 herds vaccinated for A. pleuropneumoniae and without recent respiratory problems were evaluated. One out of 118 animals tested was positive for A. pleuropneumoniae by standard culture as compared to 9 positive by coagglutination. The difference in positive results between culture and coagglutination was highly significant ( P < 0.001). Twenty-eight animals had CF titers to A. pleuropneumoniae (1:4 to ≥ 1: 128). Two hundred fifty lungs and sera samples were collected from 7 herds which had recently experienced varying degrees of respiratory disease. Thirty-nine lungs were positive for A. pleuropneumoniae by culture and 182 were positive by coagglutination. The number of positives detected by coagglutination was significantly different ( P < 0.001) from the number positive by culture. There were 172 animals with antibody titers ranging from suspect to ≥ 1:128. There were significantly fewer positive animals detected by standard culture than with the CF test ( P < 0.001). There was no significant difference between coagglutination results and CF titers when a titer of 1:4 was used as the positive threshold.
Computer Analysis as an Aid in Identification of Mycobacteria
Authors: Jarnagin Jerald L. , Swanson Merrill R.
Published: May 2011

One hundred eighty-four isolates representing 23 species of mycobacteria were identified using computer-assisted analysis. All isolates were examined using a standard series of 12 biochemical tests. These tests were selected because of their reproducibility and ease of performance in the laboratory. Data from these tests were analyzed by a computer that had been previously programmed to process the information and make a species determination. Identifications from the probability model were compared to identifications from conventional methods. There was 96.7% agreement between the 2 methods. The computer-assisted data analysis for identification provides increased accuracy over conventional methods because a statistical probability is applied. It also requires less time. Differences in computer data between mycobacterial species are discussed.
Detection of Bovine Viral Diarrhea Virus by Spot Hybridization with Probes Prepared from Cloned cDNA Sequences
Authors: Potgieter Leon N. D. , Brock Kenny V.
Published: May 2011

A cytopathic strain of bovine viral diarrhea virus (BVDV) was purified from infected cell culture fluids by isopycnic density-gradient centrifugation. Genomic RNA was extracted and tailed with adenine residues at the 3′ end with poly-A polymerase. Double-stranded complementary DNA (cDNA) was synthesized, using the poly-A-tailed RNA as a template and oligo-dT as a primer, and then cloned into the pUC9 plasmid. Virusspecific cDNA sequences, varying in length from 0.5 to 2.5 kilobases (kb), were obtained. One BVDV-specific sequence of cloned cDNA, 1.1 kb in length and with an internal Pst I restriction endonuclease cleavage site, was selected for use as a probe. The cloned cDNA insert was removed from the plasmid either with or without flanking plasmid sequences and labeled with 32P-nucleotides by nick translation for use as hybridization probes for BVDV. The performance of probes of smaller fragments of the insert was compared to that of the intact sequence in hybridization assays. In addition, 2 methods of specimen preparation were compared to establish optimum parameters for hybridization. The hybridization assay was 10–100 times more sensitive than infectivity assays for BVDV in infected cell cultures. Freezing of specimens reduced by lo-fold the sensitivity of hybridization for BVDV target sequences. The probes prepared from the cloned cDNA hybridized with all cytopathic and noncytopathic BVDV strains tested but not with uninfected cell cultures, cellular ribosomal RNA, bovine coronavirus, bluetongue virus, or bovine adenovirus 3. Probes prepared with native plasmid DNA did not hybridize with BVDV or uninfected cell cultures. Probes of 1.1 kb, 0.6 kb, and 0.5 kb hybridized with equal sensitivity to target sequences in the dot blot system. The presence of flanking plasmid sequences did not improve the hybridization performance of the probes. Hybridization results with specimens prepared by RNAsin and heat treatment were similar to those obtained after Nonidet P-40 and formaldehyde treatment.
Use of an Immunoperoxidase Test for the Detection of Bovine Herpesvirus-1 in Aborted Fetal Tissue
Authors: Smith George H. , Collins James K. , Carman Jane
Published: May 2011

An indirect immunoperoxidase (IP) procedure using a specific monoclonal antibody and an avidinbiotin-peroxidase complex was developed and applied to detect virus antigen in formalin-fixed, paraffin-embedded tissue sections. This IP procedure was compared with currently used diagnostic tests for detection of virus-induced abortions caused by bovine herpesvirus-1 (BHV–1). The IP procedure was applied to detect BHV-1 antigen in sections of liver and lung from 87 aborted fetuses. Sixteen of these cases were positive for viral antigen by IP staining. Sections from both liver and lung were positive in 15 of the 16 cases. A fluorescent antibody test (FA), which was applied to acetone-fixed frozen sections of liver and lung, gave positive results on 12 of the 87 fetuses, 11 of which were also positive by IP. Seven of the 12 FA-positive cases were positive on both sections of liver and lung. When FA and IP were compared, FA had a sensitivity of 67% and IP had a sensitivity of 94%. Virus was isolated from one of the 67 cases tested. The tissues in which antigen was most frequently detected by IP were liver, lung, and kidney. Distinct multifocal staining was seen in positive sections of all these tissues.
Serologic, Virologic, and Histopathologic Observations of Encephalomyocarditis Virus Infection in Mummified and Stillborn Pigs
Authors: Kim Hyun S. , Joo Han S. , Bergeland Martin E.
Published: May 2011

Stillborn and mummified swine fetuses from swine farms experiencing reproductive problems were investigated for evidence of infection with encephalomyocarditis (EMC) virus by fetal serology, virus isolation, and histopathologic examination. Fetal sera or thoracic fluids of 478 abnormal fetuses collected during January through December 1987 were tested for the presence of antibody specific to EMC virus. Of 478 samples tested, 175 (36.6%) had EMC virus serum neutralizing antibody titers of 1:64 or greater. The samples positive for EMC virus antibody were obtained from 38 swine farms located in 9 states in the United States. In addition to serologic observations, tissue samples of some abnormal fetuses were examined for the presence of virus and histopathologic lesions. The EMC virus was isolated in 1 case from the fetuses of an aborted litter. The isolate was serologically identical to a reference EMC virus. Nonsuppurative encephalitis and myocarditis were observed in the fetal samples collected from 2 different herds. Thoracic fluid of 1 stillborn pig with lesions was positive for EMC virus antibody at a titer of 1:512. We suggest that a widespread reproductive problem recently experienced in several major swine-producing areas of the United States may have been caused by EMC virus infection.
Quantitation of Canine Distemper Virus and Antibodies by Enzyme-Linked Immunosorbent Assays Using Protein A and Monoclonal Antibody Capture
Authors: Potgieter Leon N. D. , Ajidagba Peace A.
Published: May 2011

Monoclonal antibodies produced from 19 cloned hybridomas were selected for this study. Specific canine distemper virus (CDV) antibodies in medium from cloned hybridomas were detected by direct enzyme-linked immunosorbent assays (ELISA) and by indirect immunofluorescence. Three different sandwich ELISA systems were developed either to detect CDV in cell cultures and clinical specimens or to detect specific antibody in canine sera. Protein A and monoclonal antibodies attached in sequence to a solid phase constituted the capture system in the assays. Viral antigens were detected by sandwiching extracts of clinical specimens (or infected cell cultures), monoclonal antibody, and peroxidase-labeled protein A in sequence onto the capture layer. In 1 procedure, biotin-labeled antibody and peroxidase-labeled avidin were used as the last 2 layers in the assay. The CDV antibodies in dog sera were quantitated in a similar manner, but the sequential sandwiching levels consisted of partially purified CDV, serum specimen, and peroxidase-labeled protein A, respectively. The procedures were specific and highly sensitive.
Evaluation of Vitamin K3 Feed Additive for Prevention of Sweet Clover Disease
Authors: Casper Howard H. , Alstad Arnold D. , Tacke David B. , Johnson Ladon J. , Lloyd W. Eugene
Published: May 2011

Sweet clover poisoning in cattle is caused by an anticoagulant (dicumarol) that is formed in moldy sweet clover hay. Previous experiments with vitamin K3 and vitamin K3 in therapy trials indicated that vitamin K3 was effective in reducing prothrombin times but vitamin K3 was not. As a possible alternative in the use of toxic sweet clover hays, vitamin K3 was evaluated to see if it would prevent hemorrhagic crises when fed to cattle consuming toxic sweet clover hay. Vitamin K3 levels of 0, 0.45, 4.5, 11, and 45 mg/kg body weight/day were fed to 173-235-kg steers consuming toxic (40–50 ppm dicumarol) sweet clover. The 45-mg K3/kg/day supplement was not palatable and had to be discontinued. The 0.45, 4.5, and 11-mg K3/kg/day supplements did not significantly reduce the prothrombin times as compared to the 0-mg K3/kg/day group.
Detection of Bovine Herpesvirus-1-Specific IgM Using a Capture Enzyme Immune Assay with Isotype-Specific Monoclonal Antibodies
Authors: Osorio Fernando , Srikumaran Subramaniam , Rhodes Marvin , Christensen David , Srikumaran Pushpa
Published: May 2011

The detection of virus-specific immunoglobulin M (IgM) antibodies in acute-phase serum samples offers the possibility of making an accurate and rapid serologic diagnosis. We have developed a solid-phase capture assay that uses murine monoclonal antibodies specific for bovine IgM to separate the whole IgM fraction of a bovine serum sample. The IgM specific for bovine herpesvirus-1 (BHV-1) is then detected by the addition of viral antigen, which in turn is detected by BHV-1-specific monoclonal antibodies conjugated to horseradish peroxidase. A BHV-1 IgM antibody response was detected during the early postinfection period (7–40 days PI). Bovine herpesvirus-1 IgM antibody was not detected in sera taken from 3 animals following dexamethasone-induced viral reactivation. This method compares favorably with viral isolation, antigen detection in the clinical samples, and paired serology in the diagnosis of BHV-1 infection at a herd level.
A Rapid Monoclonal Immunofluorescence Assay for Chlamydia Psittaci in Fecal Smears from Psittacine Birds
Authors: Woods Leslie W. , Dotson Jill F. , Castro Anthony E.
Published: May 2011

One hundred two fecal specimens from psittacine birds submitted to Veterinary Laboratory Services of the California Department of Food and Agriculture at Petaluma were screened for Chlamydia psittaci by a direct immunofluorescence assay using a fluorescein-labeled monoclonal antibody conjugate specific for Chlamydia sp. Results were compared with those obtained by isolation of Chlamydia in cultures of McCoy mouse cells. The relative specificity of the direct fluorescent antibody test on fecal smears was 98.9% and the relative sensitivity was 62.5%. The results of this study suggested that the direct fluorescent antibody test was highly specific, and it proved to be a useful same-day antemortem diagnostic test for birds with symptomatic Chlamydial infection. The use of centrifugation in the cell culture assay was found to significantly enhance the level of Chlamydial infection in cell culture.
A Serological Survey of Ehrlichia Canis, Ehrlichia Equi, Rickettsia Rickettsii, and Borrelia Burgdorferi in Dogs in Oklahoma
Authors: Rodgers Sandra J. , Morton Rebecca J. , Baldwin Charles A.
Published: May 2011

Serum samples from 259 dogs were tested for antibodies to Ehrlichia canis, Ehrlichia equi, Rickettsia rickettsii, and Borrelia burgdorferi using the indirect fluorescent antibody test. The sera were obtained from submissions to the Oklahoma Animal Disease Diagnostic Laboratory during a 14-month period from June 1986 through July 1987. The rate for positive antibody titers to E. canis was 53%, to E. equi was 33%, to R. rickettsii was 38%, and to B. burgdorferi was 18%. Higher percentages of sera serologically positive to E. canis were found in the spring through the fall months, but there were no seasonal variations for E. equi, R. rickettsii, and B. burgdorferi. There was no consistent pattern of titers to the 4 antigens when age-groups of the dogs were compared. Forty-four different breeds were tested.
Serum Biochemical and Hematologic Values of Normal and Mycobacterium Bovis-Infected American Bison
Authors: Miller Lyle D. , Thoen Charles O. , Throlson Kenneth J. , Himes Elmer M. , Morgan Ronald L.
Published: May 2011

Hematologic and serum biochemical tests were used to monitor the health of 3 groups of bison in an experimental study of tuberculosis. Bison were randomly assigned to Mycobacterium bovis-infected, M. bovis -sensitized, and uninfected control groups. Hematologic measurements included total and differential leukocyte counts, hemoglobin (Hb), packed cell volume (PCV), fibrinogen, and plasma proteins. Biochemical tests included serum urea nitrogen, creatinine, aspartate amino transferase, sorbitol dehydrogenase, serum calcium, and serum phosphorus.

There were no significant differences (P = 0.05) in any test values between groups of bison. The bison data were combined and compared to similar data of cattle. The mean values for PCV and Hb were higher than values (PCV 24–46%, Hb 8–15 g/dl) for cattle. Mycobacterium bovis -infected bison had a slight increase in the number of blood monocytes and lymphocytes when compared to the uninfected bison but were within the normal ranges for bison and cattle. Other hematologic parameters were within normal ranges reported for cattle. Creatinine levels in all bison were above the normal range (1.0–1.5 mg/dl) for cattle. Phosphorus levels for M. bovis-infected and M. bovis -sensitized bison exceeded the normal range (5.6–8.0 mg/dl) reported for cattle. The level for uninfected bison was near the upper limit of normal for cattle. Mean values for other serum biochemical tests were within the normal ranges reported for cattle.
Control of Johne's Disease in Four Commercial Dairy Herds in Iowa
Authors: Thoen Charles O. , Moore Larry A.
Published: May 2011

A 6-year study was conducted in 4 dairy herds in Iowa in which Johne's disease was diagnosed previously. Fecal specimens were collected at 6-month intervals from animals 2 years of age and over for mycobacteriologic examination. Serum samples were obtained at 3-month intervals and tested by enzyme-linked immunosorbent assay (ELISA). The antigen used in the ELISA was a potassium chloride extract of a field strain of Mycobacterium paratuberculosis. The ELISA reactions were observed in 87% of the cows from which M. paratuberculosis was isolated. Dairy producers that participated in the Johne's control program reported reduced economic losses. Increased income was attributed to improved milk production, increased value of vaccinated animals sold as replacements to other dairy herds in which Johne's disease had been diagnosed, and the increased market value of slaughter animals removed from the herd.
Susceptibility of a Fetal Tongue Cell Line Derived from Bighorn Sheep to Five Serotypes of Bluetongue Virus and its Potential for the Isolation of Viruses
Authors: Castro Anthony E. , Montague Steven R. , Dotson Jill F. , Jessup David A. , DeForge James R.
Published: May 2011

A cell line (BHFTE) was derived from a tongue explant of a bighorn sheep fetus (Ovis canadensis nelsoni). The cells have been maintained through 23 serial passages, and the modal number of chromosomes was calculated to be 55. Monolayer cultures were shown to be susceptible to various viruses, including bluetongue virus (BTV). Of 5 BTV serotypes (2, 10, 11, 13, and 17) tested, each produced a cytopathic effect (CPE) on initial passage at 33 C. A field isolate (serotype 10) of BTV from a black-tailed deer (Odocoileus hemionus columbianus) in its second passage in Vero-M cells also produced CPE when inoculated into BHFTE cells. Antigens of BTV were demonstrated by direct immunofluorescence in the cytoplasm of BHFTE cells inoculated with homogenates of chicken embryos injected with clinical specimens from a domestic sheep and an Arabian oryx (Oryx gazella leucoryx).

A suspension of BTV-infected gnats (Culicoides spp.) produced CPE and BTV-specific fluorescence on the first passage in cells inoculated with a suspension of blood from sheep experimentally infected with BTV. Additionally, selected bovine viruses induced CPE in the cells. The cell line, which is free of mycoplasma and bovine viral diarrhea virus contamination, may be useful in diagnostic medicine and research involving the ruminant species.
Cultural and Biochemical Characterization of Actinobacillus and Actinobacillus-Like Species from Ram Lambs with Epididymitis
Authors: Scanlan Charles M. , Healey Mark C. , Torres Anthony R. , Johnston Alice V.
Published: May 2011

Cellular, colonial, cultural, and biochemical characteristics of 25 field strains of gram-negative pleomorphic bacilli from rams with epididymitis were compared with Actinobacillus actinomycetemcomitans American Type Culture Collection (ATCC) strain 29522 and Actinobacillus seminis ATCC strain 15768. Three field strains were identified as A. actinomycetemcomitans, 15 as A. seminis, and 2 as Haemophilus agni; however, 5 strains (3 in group A and 2 in group B) were not identified as species in the genera Actinobacillus, Haemophilus, or Pasteurella based on the taxonomic criteria in Bergey's manual of systematic bacteriology. The 5 Actinobacillus-like organisms in groups A and B were predominantly gram-negative coccobacilli and exhibited less pleomorphism than the 2 Actinobacillus species. The colonial morphologies of groups A and B were similar to the 2 Actinobacillus species but were smaller in diameter and had a pale yellow color. Groups A and B, like the actinobacilli, were facultative anaerobic and capnophilic, did not grow on MacConkey agar, and were catalase-positive and oxidase-positive. Group A reduced nitrate but group B did not. The A. seminis strains utilized ornithine, and group A utilized arginine; but group B did not utilize either omithine or arginine. All strains failed to utilize lysine or tryptophane. All strains produced acid but no gas from glucose, and the utilization of other carbohydrates varied markedly both between and within the 5 groups of bacteria. Based on carbohydrate utilization profiles, the 15 field strains of A. seminis represented 13 biotypes, but with a commercial semi-quantitative enzyme system only 1 biotype was identified. All the A. actinomycetemcomitans, A. seminis, and group B strains produced moderate to high levels of alkaline phosphatase and acid phosphatase, but the group A strains were negative. Groups A and B were negative for beta-glucuronidase, but all the A. actinomycetemcomitans and A. seminis strains produced this enzyme.
Canine Pythiosis - Isolation and Identification of Pythium Insidiosum
Authors: Bentinck-Smith John , Padhye Arvind A. , Maslin William R. , Hamilton Carolyn , McDonald Robert K. , Woody Benny J.
Published: May 2011

Pythium insidiosum was isolated from the subcutaneous tissue of a 1-year-old tan crossbreed dog and from the intestinal tract of an 18-month-old Samoyed male. Gomori's methenamine silver stain was superior to hematoxylin and eosin in demonstrating the organism in tissue sections. The agent was identified as P. insidiosum by zoospore formation in an aqueous yeast extract solution containing grass blades. Exoantigens produced in culture were shown to be identical to known P. insidiosum antigens by microimmunodiffusion.
Evaluation of an Enzyme Immunoassay for Detection of Chlamydia Psittaci in Vaginal Secretions, Placentas, and Fetal Tissues from Aborting Ewes
Authors: Sanderson Thomas P. , Andersen Arthur A.
Published: May 2011

A commercially available enzyme immunoassay (EIA) for the detection of Chlamydia trachomatis in human urogenital and conjunctival specimens was compared with isolation in cell culture for the detection of Chlamydia psittaci in vaginal and placental swabs from aborting ewes and swabs of aborted fetal tissues. The EIA on vaginal swabs collected from 10 ewes experimentally infected with C. psittaci had a sensitivity of 85.7% and a specificity of 85.7%. Vaginal swabs collected at the time of abortion or within 3 days were the best samples for detection of Chlamydial infection. The 29 vaginal swabs collected during this period from experimentally infected ewes were all strongly EIA-positive, and Chlamydia were isolated from 28. The EIA on vaginal swabs from 78 field cases of abortion had a sensitivity of 78.0% and a specificity of 76.8%. The EIA on swabs of cotyledons from 65 placentas had a sensitivity of 100% and a specificity of 75.0% compared with isolation in cell culture. The EIA on 57 swabs of fetal tissues or body fluids from 10 aborted fetuses or weak lambs from experimentally infected ewes had a sensitivity of 26.6% and a specificity of 88.1% compared with isolation in cell culture. Limitations of the EIA are discussed.
A Blocking ELISA for the Detection of Specific Antibodies to Bovine Respiratory Syncytial Virus
Authors: Rhodes Marvin B. , Klucas Carol A. , Frey Merwin L. , Anderson Gary A.
Published: May 2011

A blocking enzyme-linked immunosorbent assay (ELISA) has been adapted to detect specific antibodies in bovine sera to respiratory syncytial virus using a horseradish peroxidase-labeled monclonal antibody to the fusion protein of the virus. This assay plus an indirect blocking ELISA and indirect ELISA were used to detect antibodies to the bovine respiratory syncytial virus (BRSV) in 159 field-origin bovine sera. Results of these assays were compared with serum antibody titers measured by the serum neutralization (SN) test. Over a 56-day period, the mean neutralization titers and the mean delta absorbance values for the blocking ELISA, on the same sera, showed similar declines. However, the calculated correlation coefficients between mean SN titer and mean absorbance value for the blocking ELISA of the individual sera ranged from −0.2 to −0.5 depending on the source of sera. Similar values were obtained whether using crude or purified viral antigen in the assays. Corresponding calculated correlation coefficients were generally higher for the indirect blocking ELISA or indirect ELISA than for the blocking ELISA. The blocking ELISA was between 70 and 64% as sensitive as the serum neutralization test with a specificity of 100 or 90% using the crude and purified viral antigen, respectively. The indirect blocking ELISA and indirect ELISA had similar calculated sensitivities and specificities. The blocking ELISA was faster to run than either of the other ELISA's or the neutralization test. Further, nonspecific background absorbance was obviated because the blocking ELISA detects antibodies to 1 specific viral protein, the fusion protein. These studies suggest that the blocking ELISA should be useful as a serological test for BRSV antibodies.
Use of in Situ Hybridization for the Detection of Foot-and-Mouth Disease Virus in Cell Culture
Authors: Meyer Richard F. , Brown Corrie C. , Molitor Thomas W. , Vakharia Vikram N.
Published: May 2011

Biotinylated complementary DNA (cDNA) and RNA probes were prepared from a specific and highly conserved section of the foot-and-mouth disease virus (FMDV) genome coding for the RNA-dependent RNA polymerase. Hybridization was conducted on FMDV-infected, bovine enterovirus (BEV)-infected, and noninfected swine kidney cell cultures. The detection system utilized the enzyme system streptavidin-alkaline phosphatase, the substrate phosphate, and the chromogen nitroblue tetrazolium. Intense cytoplasmic granular staining was present at 2 and 4 hr postinfection (hpi), with less staining observed at 24 hpi. The staining was specific for FMDV, as indicated by a lack of staining of noninfected cells and BEV-infected cells. With the RNA probe, positive cells were detected up to the highest viral dilution assayed, which was approximately 96 TCID50. The cDNA probe was slightly less sensitive, detecting positive cells at lo-fold lower dilutions. This technique could prove useful in the diagnosis of foot-and-mouth disease in animals or in the detection of FMDV in biologics submitted for importation.
Rabies-Like Inclusions in Dogs
Authors: Nietfeld Jerome C. , Rakich Pauline M. , Tyler David E. , Bauer Rudy W.
Published: May 2011

Inclusion bodies, indistinguishable from rabies inclusion bodies (Negri bodies), were found in the brains of 8 nonrabid dogs. The inclusions were compared to Negri bodies present in neurons of rabies-positive animals and examined for the presence of rabies virus by a combination of immunoperoxidase staining (7 cases), fluorescent antibody (FA) staining (1 case), and transmission electron microscopy (4 cases). Positive immunoperoxidase staining for rabies was obtained in brain tissues from FA rabies-positive animals. All brain tissues from the 7 dogs stained by the immunoperoxidase method and the brain from the 1 dog stained by the FA method were negative for rabies. Rabies virus was not found in inclusion-containing neurons in the cases examined by transmission electron microscopy. These results emphasize the importance of FA testing and mouse inoculation for the diagnosis of rabies.
Feline Immunodeficiency Virus Infection in a Population of Pet Cats from Southeastern Florida
Authors: Fisch Harvey , Altman Norman H.
Published: May 2011

Blood samples from 95 randomly selected pet cats that were brought to veterinarians in southeastern Florida were tested for antibodies to feline immunodeficiency virus (FIV). Virus-specific antibodies (indicative of virus infection) were found in 8 of the 95 (8.4%) cats tested. All of the virus-infected cats were males (statistically significant, P ≤ 0.016) and were at least 1 year of age. The 3 most severely ill cats infected with FIV were also infected with feline leukemia virus.
Element Concentrations in Livers and Kidneys of Ranch Mink
Authors: Stejskal Susan M. , Aulerich Richard J. , Slanker Michael R. , Braselton W. Emmett , Lehning Ellen J. , Napolitano Angelo C.
Published: May 2011

Reference ranges for element concentrations in livers and kidneys of “healthy” mink of known age, sex, and coat color and fed a conventional diet were determined. After euthanasia and removal of the pelts, liver and kidney samples were collected from 174 mink and analyzed for 22 elements using inductively coupled argon plasma emission spectroscopy. The diet of the mink was also analyzed for element concentrations. Descriptive statistics of element concentrations for livers and kidneys of the mink are given and compared with dietary element concentrations