bluuebunny/crossref_metadata_2025 huggingface dataset:
A Toxicological Investigation of the Garden Shrub Brunfelsia Calcyina var. Floribunda (Yesterday-Today-and-Tomorrow) in Three Species
Authors: Spainhour Charles B. , Fiske Robert A. , Flory Wayne , Reagor John C.
Published: May 2011

Brunfelsia calcyina var. floribunda is an ornamental evergreen shrub found in the United States. A diagnosis of the fatal intoxication of a canine due to consumption of plant material (primarily berries) was made. The significant features of the clinical constellation were similar to those seen with substances interfering with the neurotransmission process, such as lathyrus or strychnine. Necropsy findings on the canine were unrevealing. Toxicologic studies performed on mice and rats with ground shrub material demonstrated that all parts of this plant are toxic, but unequally so. All plant preparations produced signs similar to those of a spinal convulsant. There were no distinguishing gross pathologic or histopathologic findings associated with the toxicoses induced in the laboratory animals with preparations from this plant. The toxic principles from this shrub are water soluble and very stable. The ability of aqueous extracts stored at 4 C to produce the clinical syndrome and subsequent lethality remained unchanged over a period of 4 months. Exposures are not always fatal. They most often occur in the canine and there is a significant hazard for small children.
The Evaluation of Postmortem Ocular Fluid Analysis as a Diagnostic Aid in Sows
Authors: Drolet Richard , D'Allaire Sylvie , Chagnon Madeleine
Published: May 2011

This study was undertaken to evaluate the diagnostic usefulness of postmortem ocular fluid analysis in estimating the antemortem status of various serochemical constituents. Chemical values of serum and aqueous and vitreous humors were compared following different procedures. A blood sample and the 2 eyes were collected from each of 100 sows at a nearby abattoir. The results obtained from immediate centrifugation of ocular fluids after sampling were compared with those samples in which centrifugation was delayed by 2 hours. Two different postmortem intervals were used for sampling ocular fluids, 2 and 24 hours. Concentrations of urea, calcium, phosphorus, potassium, sodium, and chloride were determined from serum and humors. Delayed centrifugation did not affect chemical values of ocular fluids nor the relationships between serum and humors. Phosphorus and potassium values increased significantly with the postmortem interval in both aqueous and vitreous humors. The relationships between chemical values of ocular fluids and serum were determined using simple linear regression. There was a poor correlation between ocular fluid and serum values for all electrolytes; a significant correlation was found only for urea concentrations in both humors.
A Study Comparing the Immunologic Responses of Swine to Pseudorabies Viral Antigens Based on the ELISA, Serum Virus Neutralization, and Latex Agglutination Tests
Authors: Schoenbaum Mark A. , Beran George W. , Murphy Dorothy P.
Published: May 2011

A study of pseudorabies virus (PRV)-vaccinated pigs comparing the immune responses detected by the latex agglutination test (LAT) with responses detected by other routine tests for pseudorabies antibodies indicated that LAT was more sensitive than either the enzyme-linked immunosorbent assay (ELISA) or the serum virus neutralization test (SVNT). The LAT detected antibodies sooner than ELISA and SVNT in unvaccinated pigs after challenge with virulent PRV. The specificities of the 3 tests were found to be near 100%. The LAT is a good alternative to SVNT or ELISA for detection of PRV-specific antibodies.
Laboratory Diagnosis of African Horse Sickness: Comparison of Serological Techniques and Evaluation of Storage Methods of Samples for Virus Isolation
Authors: House Carol , Mikiciuk Peter E. , Beminger Mary Lou
Published: May 2011

Five serological methods of diagnosing African horse sickness were evaluated, using a battery of serum samples from experimental horses vaccinated and challenged with each serotype of African horse sickness virus (AHSV1 through AHSV9): agar gel immunodiffusion (AGID), indirect fluorescent antibody (IFA), complement fixation (CF), virus neutralization (VN), and enzyme-linked immunosorbent assay (ELISA). The 5 tests were also compared using a panel of field samples, convalescent equine sera with antibodies to domestic equine viral diseases, and sera from horses awaiting export. The ELISA described in this paper was group specific. It did not require calibration with a standard positive serum but did yield elevated values with negative sera that were repeatedly frozen and thawed or heat inactivated. The IFA test was sensitive but could not be used on some field sera as the control cells exhibited fluorescence, possibly due to the animal being recently vaccinated with cell culture material. Sixty-two experimental sera were compared by VN, CF, AGID, and ELISA. Forty sera, 10 positive and 30 negative, were correctly classified by the 5 serologic assays. The 22 remaining sera gave mixed reactions. The AGID had no false positive results but had false negative results for up to 20% of the samples, depending upon the comparison. The VN, CF, and ELISA were similar in their variability.

The length of time that virus could be recovered from a viremic blood sample was compared in an evaluation of storage methods for virus isolation samples. Washed erythrocytes were held at 4 C, washed erythrocytes plus stabilizer were held at −70 C, and blood that was drawn into a preservative (oxalate/phenol/glycerol) was held at 4 C. Virus was isolated for 12 months from a sample stored as washed erythrocytes at 4 C but for only 6 months from the same blood stored by the other 2 methods.
Ovine Fetal Telencephalic White Matter Necrosis
Authors: Montgomery Donald L. , Gilmore William C.
Published: May 2011

Discrete foci of white matter necrosis were observed in the telencephalic white matter in 6 casesof spontaneous ovine abortion. All fetuses were in the third trimester of gestation and 2 of the 6 were term fetuses. The lesions were distributed randomly throughout the cerebrocortical white matter and consist of pale hypocellular plaques with a variable glial cell response and encrustation of necrotic cellular processes. A variety of additional lesions were present in the brains of the aborted fetuses. The lesions of telencephalic white matter necrosis have not been reported previously and are unlike those associated with known viral infections of the developing fetus. Based on the character of the lesions, a hypoxic-ischemic or hypotensive mechanism is suggested. The lesions are compared with a similar condition in human fetuses and neonates.
The Effect of Storage Time on Isolation of Leptospira Interrogans from Bovine Kidneys
Authors: Miller David A. , Wilson Mark A. , Beran George W.
Published: May 2011

Kidney tissues from 20 cattle infected with Leptospira interrogans serovars hardjo, pomona, or grippotyphosa were cultured on the day of slaughter and 3, 6, and 8 days later to examine the effect of storage time on the recovery of leptospires by conventional culture methods. Leptospires were isolated from 85% of infected bovine kidney tissues cultured on day 1, and from 95%, 90%, and 90% of kidney tissues stored in transport medium at 4 C for 3, 6, and 8 days, respectively, prior to inoculation of culture media.
Limits of Detection of Bluetongue Virus with Different Assay Systems
Authors: Wechsler Sally J. , Austin Kathleen J. , Wilson William C.
Published: May 2011

The sensitivity of different assay systems for detecting low concentrations of bluetongue virus (BTV) were compared. These assays included blind passage on baby hamster kidney (BHK-21) cells and on cattle pulmonary artery endothelial (CPAE) cells, immunoperoxidase staining of cells on multiwell slides, and cDNA/RNA hybridization of BTV infected cells. Nine serial lo-fold dilutions of a cell culture-adapted BTV serotype 11 were tested (each dilution was treated as a separate sample) in all assays. Visual inspection for cytopathic effects (CPE) during 3 passages in BHK-21 cells detected samples that contained ≥ 3 plaque forming units (PFU)/ml of BTV. Evidence of CPE during 3 passages in CPAE cells detected samples that contained ≥ 0.3 PFU/ml of BTV. A limit of detection (≥0.3 PFU/ml) was obtained faster by immunoperoxidase staining of BTV-inoculated CPAE cells on multiwell slides and incubated for 3 days. The cDNA/RNA hybridizations of CPAE and BHK-21 cells incubated for 2 or 3 days, respectively, with BTV dilution samples detected samples that contained ≥30 PFU/ml. Of the assay systems examined, immunoperoxidase staining of CPAE cells on multiwell slides inoculated with cell culture-adapted BTV was the most sensitive and fastest assay for definitive virus identification.
The Isolation of Lumpy Skin Disease Virus and Bovine Herpesvirus- from Cattle in Egypt
Authors: House James A. , Wilson Terrance M. , Nakashly Sinan El , Karim Ikram A. , Ismail Ibrahim , Danaf Nabil El , Moussa Aly M. , Ayoub Nazmi N.
Published: May 2011

Lumpy skin disease (LSD) virus (LSDV) was isolated for the first time from cattle in Egypt in 2 disease outbreaks. Bovine herpesvirus-4 (BHV-4) and LSDV were detected in a pooled sample from the first outbreak (Suez). Only LSDV was isolated from the second outbreak (Ismalia). The capripoxviruses were identified as LSDV by neutralization with specific antiserum and by their ability to produce generalized LSD in experimentally inoculated cattle.
Monoclonal Antibody in the Identification of Haemophilus Somnus
Authors: Thomson Mary S. , Lauerman Lloyd H. , Wilt Gerald R.
Published: May 2011

Electrophoretic comparisons of outer membrane proteins of Haemophilus somnus isolates revealed 2 major protein bands (46 and 14 kilodaltons [kD]) common to all isolates tested. A monoclonal antibody raised against H. somnus reacted to the 46-kD band.

Coagglutination tests were performed using a monoclonal antibody coagglutination assay. The monoclonal reagent was produced by incubating Cowan strain Staphylococcus aureus suspension, used as a source of crude protein A, with mouse ascitic fluid monoclonal antibody or goat anti-H. somnus hyperimmune serum. Bacteria to be tested were suspended at a concentration of 4.5 × 109 cells/ml. The coagglutination test was performed by the addition of 50 μ1 of the monoclonal reagent to 50 μ1 of the bacterial suspension on a glass plate and manual rotation for 2–3 minutes.

The coagglutination assay using Cowan strain Staphylococcus aureus protein A, coupled with the monoclonal antibody, agglutinated 10 different H. somnus isolates. The antibody reagent did not coagglutinate with Actinobacillus suis, A. equuli, Pasteurella haemolytica, P. multocida, or P. pneumotropica under similar test conditions.
Some Immune Responses in Cattle Exposed to Mycobacterium Paratuberculosis after Injection with Modified-Live Bovine Viral Diarrhea Virus Vaccine
Authors: Thoen Charles O. , Waite Kyle J.
Published: May 2011

Delayed-type hypersensitivity responses to Mycobacterium paratuberculosis purified protein derivative (PPD) were decreased in cows experimentally exposed to M. paratuberculosis 7 days after exposure to a modified-live bovine viral diarrhea virus (ML-BVDV) vaccine. In vitro lymphocyte blastogenic responses to phytohemagglutinin were decreased in each of 3 cows 7 days after exposure to ML-BVDV vaccine. Also, decreased lymphocyte blastogenic responses to M. paratuberculosis PPD were observed in cultures of 2 of 3 cows 7 days after exposure to ML-BVDV vaccine. No significant differences in enzyme-linked immunosorbent assay reactions were detected in sera of M. paratuberculosis-infected cattle collected before and at 4 and 12 weeks after exposure to ML-BVDV vaccine.
Foot-and-Mouth Disease Virus in the Llama (Lama Glama): Diagnosis, Transmission, and Susceptibility
Authors: Lubroth Juan , Yedloutschnig Ronald J. , Culhane Veronica K. , Mikiciuk Peter E.
Published: May 2011

Foot-and-mouth disease virus (FMDV) was shown to be transmitted from either cattle to llamas, llamas to swine (interspecies), or llamas to llamas (intraspecies). Response to FMDV varied greatly in the 6 llamas studied; 3 llamas developed generalized clinical disease with mild pyrexia, 2 after intradermolingual inoculation, and 1 after exposure to a calf infected with FMDV serotype A24. Another contact llama developed vesicular lesions on all 4 extremities but no oral lesions. Two contact llamas, in separate study groups, did not seroconvert or develop clinical signs of FMDV infection. All 4 llamas showing clinical disease developed virus-neutralizing antibodies against FMDV A24 and antibodies against the virus-infection-associated antigen. Virus-neutralizing antibody titers remained elevated for over 200 days postinoculation or exposure. Antibodies to virus-infection-associated antigen were detected several days after virus-neutralizing antibody appeared and became weaker 100–125 days post-FMDV exposure in 3 of the 4 clinically affected llamas. One inoculated llama was still positive for virus-infection-associated antigen at 360 days after inoculation. Foot-and-mouth disease virus A24 was not detected from esophageal-pharyngeal fluid specimens beyond 8 days postexposure using in vitro techniques.
Laboratory Findings Associated with Abomasal Ulcers/Tympany in Range Calves
Authors: Mills Kenneth W. , Johnson Jerre L. , Jensen Rue L. , Woodard Lynn F. , Doster Alan R.
Published: May 2011

The etiology of abomasal ulcers/tympany was investigated in 48 animals from 36 ranches in Wyoming and Nebraska. Results indicate that subclinical trace mineral deficiencies of copper and/or selenium exist in the range cattle in west central Nebraska and Wyoming. Etiological agents most frequently incriminated by bacteriologic cultures and/or histopathic examination were Clostridium perfringens and Campylobacter species. Histopathologic evaluation of abomasums revealed 31 of 38 cases contained abundant gram-positive bacteria associated with the damaged abomasal mucosa. Campylobacter-like organisms were demonstrated in 9 of 38 cases using the modified Dieterle stain. Clostridium perfringens was isolated in 14 of 38 cases, and Campylobacter jejuni was recovered from 5 of 38 cases.
Pulmonary Edema and Hydrothorax in Swine Produced by Fumonisin B1, a Toxic Metabolite of Fusarium Moniliforme
Authors: Harrison Lenn R. , Colvin Billy M. , Greene J. Todd , Newman Louis E. , Cole John R.
Published: May 2011

Pulmonary edema and hydrothorax were observed in mature swine that died approximately 5 days after consuming corn screenings. These postmortem observations were reproduced in younger swine (16–24 kg) that died within 1 week when fed the corn screenings under experimental conditions. Additionally, pulmonary edema and hydrothorax occurred in a pig (7.1 kg) that died after receiving 4 daily intravenous injections of fumonisin B,. A fungus was isolated from the corn screenings that is identical to Fusarium moniliforme MRC-826 in colony morphology and under microscopic examination.
A Grading System for Lymphocytic Plasmacytic Colitis in Dogs
Authors: Roth Lois , Walton Amelia M. , Leib Michael S. , Burrows Colin F.
Published: May 2011

Colonic mucosal samples were obtained every 4 weeks for 13 months from 6 clinically normal dogs and from 47 dogs with a clinical diagnosis of chronic inflammatory bowel disease. All samples were graded on a scale of 0–5, based upon the quantity of lymphocytes and plasma cells in the lamina propria, epithelial changes, and the presence of ulcers and erosions. A grade of ≤2.0 was considered normal and was assigned to 77 of 78 samples from clinically normal dogs and 28 of 48 samples from dogs with diarrhea. A transient increase in cellulaiity was noted in 1 sample from 1 control dog. Nineteen dogs with clinical disease had obvious histologic abnormalities. The grading scheme described provides the pathologist with an objective criterion for the microscopic evaluation of colonic mucosal samples obtained by endoscopic techniques and offers clinicians a method of assessing the dog's progress and response to therapy.
A Sensitive Bioassay for Detection of Dietary Estrogens in Animal Feeds
Authors: Welshons Wade V. , Rottinghaus George E. , Nonneman Dan J. , Dolan-Timpe Marianne , Ross P. Frank
Published: May 2011

Estrogen-responsive proliferation in the MCF-7 cell line was used as a bioassay for detection of dietary estrogens. The bioassay procedure was adapted to screen for estrogenic activity in feedstuffs that have been associated with hyperestrogenism in livestock. Methanolic feed extracts were added to the cell culture medium at μ1/ml concentrations for 4 days, after which the cell proliferation response was measured as DNA content. The half-maximal response for estradiol occurred at 2 pM, or 0.54 pg/ml. For zearalenone, a weaker estrogen, the half-maximal response occurred at approximately 200 pM, or 64 pg/ml. The bioassay was calibrated against a number of known estrogens (estradiol, diethylstilbestrol, zearalenone, zearalanol [cattle implant], β-zearalenol, zearalane), including the naturally occurring phytoestrogens (formononetin, genistein, daidzein, biochanin A, and coumestrol). The estrogenic activity of feed samples was expressed as equivalents of zearalenone (ppm zearalenone) that would have to be present to equally stimulate proliferation of the MCF-7 cells. The sensitivity of the bioassay was 0.05–0.1 ppm equivalents of zearalenone in feed, well below the threshold level associated with reproductive problems. The feed additive melengestrol acetate (MGA) showed no estrogenic activity in this assay. Estrogenic activity of feed extracts was confirmed by competitive inhibition with the antiestrogens tamoxifen or LY156758 (keoxifene) to show that stimulation of growth by feed extracts was through an estrogenic mechanism. Confirmation of known estrogens was by tandem mass spectroscopy. The assay is a sensitive and reliable screening procedure for detecting estrogenic activity in feedstuffs.
Immunohistochemical Identification of Infectious Pancreatic Necrosis Virus in Paraffin-Embedded Tissues of Atlantic Salmon (Salmo Salar)
Authors: Evensen Øystein , Rimstad Espen
Published: May 2011

Infectious pancreatic necrosis virus serotype Sp was identified by immunohistochemistry in formaldehyde-fixed and paraffin-embedded tissue of Atlantic salmon ( Salmo salar). The immunoreaction was present in degenerating and necrotic cells in exocrine pancreatic cells. Cross reactions were observed with rabbit antisera against serotypes Sp, Ab, and VR-299 in neutralization tests and western blotting. Immunohistochemically, only Sp antiserum produced positive immunostaining to Sp antigens, whereas antisera to serotypes Ab and VR-299 were negative.
Limitations of in Situ Hybridization for the Detection of Bluetongue Virus in Blood Mononuclear Cells
Authors: Dangler Charles A. , de la Concha-Bermejillo Andrés , Stott Jeffrey L. , Osburn Bennie I.
Published: May 2011

In situ nucleic acid hybridization was tested for the ability to detect bluetongue virus (BTV) nucleic acids in blood mononuclear cells. A standard protocol was devised and applied to the demonstration of BTV genetic sequences in cultured bovine mononuclear cells that had been infected in vitro. In situ hybridization using biotinylated single-stranded RNA probes, in the presence of 50% formamide at 50 C, demonstrated an intense, positive signal in 0.00l–0.01% of the BTV-infected cultured mononuclear cells. The protocol was applied to isolated mononuclear cells from an experimentally infected heifer. No infected cells were observed by this method, although the blood specimens were obtained during peak viremia.
Evidence for a Porcine Respiratory Coronavirus, Antigenically Similar to Transmissible Gastroenteritis Virus, in the United States
Authors: Wesley Ronald D. , Woods Roger D. , Hill Howard T. , Biwer Jerome D.
Published: May 2011

A respiratory variant of transmissible gastroenteritis virus (TGEV), designated PRCV-Ind/89, was isolated from a swine breeding stock herd in Indiana. The virus was readily isolated from nasal swabs of pigs of different ages and induced cytopathology on primary porcine kidney cells and on a swine testicular (ST) cell line. An 8-week-old pig infected oral/nasally with the respiratory variant and a contact pig showed no signs of respiratory or enteric disease. These pigs did not shed virus in feces but did shed the agent from the upper respiratory tract for approximately 2 weeks. Baby pigs from 2 separate litters (2 and 3 days old) also showed no clinical signs following oral/nasal inoculation with PRCV-Ind/89. In a third litter, 5 of 7 piglets (5 days old) infected either oral/nasally or by stomach tube developed a transient mild diarrhea with villous atrophy. However, virus was not isolated from rectal swabs or ileal homogenates of these piglets, and viral antigen was not detected in the ileum by fluorescent antibody staining even though the virus was easily recovered from nasal swabs and lung tissue homogenates. Swine antisera produced against PRCV-Ind/89 or enteric TGEV cross-neutralized either virus. In addition, an anti-peplomer monoclonal antibody, 4F6, that neutralizes TGEV also neutralized the PRCV-Ind/89 isolate. Radioimmunoassays with a panel of monoclonal antibodies indicated that the Indiana respiratory variant and the European PRCV are antigenically similar.
Evaluation of an Enzyme Immunoassay for Diagnosis of Bovine Leptospirosis Caused by Leptospira Interrogans Serovar Hardjo Type Hardjo-Bovis
Authors: Trueba Gabriel A. , Bolin Carole A. , Thoen Charles O.
Published: May 2011

Sensitivity and specificity of 4 different antigen preparations from Leptospira interrogans serovar hardjo were compared in an enzyme immunoassay for detection of antibodies against serovar hardjo type hardjo-bovis in serum. Two antigens prepared using detergents showed serogroup cross-reactivity. A mechanically extracted membrane and a lipopolysaccharide antigen showed a high degree of leptospiral serogroup specificity. The lipopolysaccharide antigen was the most suitable antigen for detection of anti-hardjo antibodies. Enzyme immunoassay was more sensitive than the microscopic agglutination test for detecting antibodies in serum from experimentally and naturally infected cattle. It was not possible to differentiate vaccinated from infected animals or to detect a secondary immune response in vaccinated animals that were subsequently infected.
Minimal Inhibitory Concentrations of Five Antimicrobials against Treponema Hyodysen Teriae and Treponema Innocens
Authors: Messier Serge , Higgins Robert , Moore Camille
Published: May 2011

The minimal inhibitory concentrations of carbadox, dimetridazole, lincomycin, ronidazole, and tiamulin against isolates of Treponema hyodysenteriae and Treponema innocens were determined by an agar-dilution method. The results obtained indicated that tiamulin was the most effective antimicrobial in vitro against T. hyodysenteriae, followed by carbadox. Dimetridazole, lincomycin, and ronidazole had poor efficacy in vitro against the T. hyodysenteriae isolates. Isolates of T. innocens were more sensitive to the various antimicrobials. Carbadox and tiamulin were the most effective in vitro, followed by ronidazole, dimetridazole, and lincomycin.
Detection of Cattle Infected with Bovine Viral Diarrhea Virus Using Nucleic Acid Hybridization
Authors: Roberts Karen L. , Collins James K. , Carman Jane , Blair Carol D.
Published: May 2011

A ribonucleic acid (RNA) hybridization assay to identify cattle infected by bovine viral diarrhea virus (BVDV) is described. The RNA probe was derived from the coding region at the 3′ end of the genome of the NADL strain of BVDV. Total RNA from infected cell cultures or peripheral blood leukocytes from suspect animals was extracted and applied to nylon membranes with a slot blot apparatus. Peripheral blood leukocytes were tested concurrently for BVDV by virus isolation. The results of hybridization and virus isolation were in agreement for 92% of the cases. When compared with virus isolation, hybridization had a sensitivity of detection of 59.5% and a specificity of 95%. Cross-reactivity to RNA extracts of border disease virus-infected cells was noted. No cross-reactivity was detected to other common bovine viruses (bovine herpesvirus-l, bovine respiratory syncytial virus, parainfluenza-3 virus, and bluetongue virus), to viruses classified in related families (equine arteritis virus and Venezuelan equine encephalitis virus), or to viruses having similar genomic organization (dengue virus type 2 and Japanese encephalitis virus).
Detection of Bluetongue Virus RNA by in Situ Hybridization: Comparison with Virus Isolation and Antigen Detection
Authors: Schoepp Randal J. , Blair Carol D. , Roy Polly , Beaty Barry J.
Published: May 2011

An in situ nucleic acid hybridization (ISH) technique was developed to detect bluetongue virus (BTV) RNA in cell culture. The sensitivity of the ISH technique was compared with virus isolation (VI) and antigen detection, using an indirect fluorescent-antibody (IFA) or an enzyme immunocytoassay (EICA) technique, for detection of 5 BTV serotypes indigenous to the United States. The VI was the most sensitive technique, detecting BTV early after infection of the cells. The IFA and EICA were of similar sensitivity; BTV antigen could be detected shortly after demonstration of virus by isolation. The sensitivity of ISH for detection of BTV-17 was equivalent to that of antigen detection. The ISH was not as sensitive as VI or antigen detection when assaying for the other BTV serotypes.
Differentiation between Transmissible Gastroenteritis Virus and Porcine Respiratory Coronavirus Using a cDNA Probe
Authors: Wesley Ronald D. , Wesley Irene V. , Woods Roger D.
Published: May 2011

A plasmid, pG3BS, containing a cDNA clone from the 5′ coding region of the peplomer glycoprotein gene appears to be specific for enteric transmissible gastroenteritis virus (TGEV) strains and for live-attenuated TGEV vaccines. This cDNA probe is used to differentiate porcine respiratory coronavirus (PRCV) isolates from TGEV field and vaccine strains by a slot blot hybridization assay. Probe pG3BS also hybridizes to canine coronavirus (CCV) RNA but does not hybridize to antigenically related feline infectious peritonitis virus (FIPV) RNA. The RNAs of 13 enteric TGEV isolates from the United States, Japan, and England, 4 US-licensed live-attenuated TGEV vaccines, and antigenically closely related CCV were detected by pG3BS. The RNAs of FIPV and 3 US isolates of PRCV did not react with pG3BS but were detected by a TGEV-derived plasmid, pRP3. Pigs infected with either PRCV or TGEV test serologically positive for TGEV antibody by the serum neutralization test. Characterization of the virus circulating in a swine herd by the pG3BS probe will differentiate between an enteric TGEV and a respiratory PRCV infection.
A Current Assessment of the Role of Porcine Parvovirus as a Cause of Fetal Porcine Death
Authors: Mengeling William L. , Lager Kelly M. , Zimmerman Jeffery K. , Samarikermani Nader , Beran George W.
Published: May 2011

One hundred one litters containing 1 or more dead porcine fetuses were collected at an Iowa abattoir during a 2-month interval and examined for evidence of viral infection. Each of 1,137 fetuses (302 dead, 835 alive) of these litters was tested for porcine parvovirus (PPV) antigens by direct immunofluorescence microscopy (FA) of fetal lung. Antigens of PPV were detected in the lungs of most of the fetuses of 11 of the litters. The 11 FA-positive litters contained 105 dead (100 FA-positive) and 14 live (12 FA-positive) fetuses. Infectious PPV was isolated from 10 of the 11 FA-positive litters and from 3 of the 90 FA-negative litters. No cytopathogenic agents other than PPV were isolated from any of the litters. Eleven of 101 (11%) litters examined and 100 of 302 (33%) dead fetuses examined were FA positive for viral antigen, indicating that PPV remains as a major cause of porcine fetal death.
Genital Bovine Papillomavirus Infection in Saudi Arabia
Authors: Elzein El Tayb Ebu , Sundberg John P. , Housawi Fadhel M. , Gameel Ahmed A. , Ramadan Ramadan O. , Hassanein Magdi M.
Published: May 2011

Genital bovine papillomavirus infection was observed for the first time in the Al-Ahsa region of Saudi Arabia. The disease involved 1 female and 2 male 2–4-year-old crossbred cattle. Fibropapillomas (warts) were limited to the prepuce and vulva. Electron micrographs of thin sections of the lesions revealed the presence of intranuclear viruslike particles. Using a broadly cross-reactive rabbit polyclonal antiserum directed against papillomavirus group-specific antigens, the infection was confirmed by immunohistochemical staining of paraffin-embedded tissues to be due to a papillomavirus. Staining with a series of monoclonal antibodies of various specificities indicated that the virus was bovine papillomavirus type 1. Attempts to propagate the virus by inoculation of tumor homogenates onto chorioallantoic membranes of chicken embryos were unsuccessful.
Hereditary Lymphedema in Hereford Cattle
Authors: Schild Ana L. , Riet-Correa Franklin , Mendez Maria C.
Published: May 2011

Congenital lymphedema is described in Hereford cattle. The disease was characterized by edema of the hind limbs, sometimes forelimbs, tail, and prepuce. Lymphatic system lesions were hypoplasia and aplasia of lymph vessels and prescapular, iliofemoral, and popliteal lymph nodes. Test matings demonstrated the transmission of the disease by an autosomal dominant trait with variable expressivity and incomplete penetrance.
Traumatic Reticulitis in Cattle: A Review of 60 Fatal Cases
Authors: Roth Lois , King John M.
Published: May 2011

Sixty fatal cases of traumatic reticulitis in cattle were reviewed. Fifty-nine cases were caused by fragments of wire. A nail perforated the reticulum of 1 animal. Common clinical signs included decreased milk production, anorexia, fever, and weight loss. Abnormal or muffled heart sounds associated with pericarditis and epicarditis was the most common sequela, occurring in 40 cases. Fibrous adhesions found at necropsy in all cases suggest that initial clinical signs are difficult to recognize and in most cases it takes weeks to months for abnormalities to be observed.
Characteristics of an Unusual Mycoplasma Isolated from a Case of Caprine Mastitis and Arthritis with Possible Systemic Manifestations
Authors: DaMassa Al J. , Nascimento Elmiro R. , Khan Mazhar I. , Yamamoto Richard , Brooks Dale L.
Published: May 2011

A mycoplasma designated strain GM790A was isolated from milk and internal organs of 2 lactating goats showing mastitis and arthritis. The isolate was not related serologically to any of the currently known ovine-caprine mycoplasmas, except an isolate designated Mycoplasma sp. G, first recorded from the external ear canal of clinically normal goats in Australia. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and restriction enzyme DNA studies of strain GM790A and Mycoplasma sp. G revealed similar but not identical patterns. The inoculation of strain GM790A into the teat canal of 2 lactating goats resulted in an abrupt diminution of lactation leading to mastitis and agalactia in about 3 days. A maximum of 1 × 107 colony-forming units (CFU) of the mycoplasma were shed per ml of mammary secretion. Milk production partially resumed at a low level 3 weeks postinoculation, the longest period tested, but the milk still contained 1 × 102 CFU of the agent. The results of this study indicate that strain GM790A possesses pathogenic potential for the goat and most probably represents a new species of the genus Mycoplasma.
In Vitro Antimicrobial Susceptibility of Pasteurella Haemolytica and Pasteurella Multocida Recovered from Cattle with Bovine Respiratory Disease Complex
Authors: Post Karen W. , Cole N. Andy , Raleigh Russell H.
Published: May 2011

The antimicrobial susceptibilities of 421 Pasteurella haemolytica and 158 P. multocida isolates recovered from cattle with respiratory disease were determined with a microdilution minimal inhibitory concentration test system. Isolates were analyzed for patterns of resistance to ampicillin, ceftiofur, erythromycin, gentamicin, penicillin, spectinomycin, sulfachlorpyridazine, sulfadimethoxine, tetracycline, and tylosin. All isolates tested were found susceptible to ceftiofur and sulfachlorpyridazine. Pasteurella haemolytica isolates were resistant to ampicillin, penicillin, sulfadimethoxine, tetracycline, and tylosin. Pasteurella multocida isolates were resistant to sulfadimethoxine, tetracycline, and tylosin.
Comparison of in Vitro Replication and Cytopathology Caused by Strains of Canine Distemper Virus of Vaccine and Field Origin
Authors: Evans Mary B. , Bunn Thomas O. , Hill Howard T. , Platt Kenneth B.
Published: May 2011

Three biological properties of canine distemper virus were examined to determine if any would consistently differentiate field from vaccine strains of the virus. The properties were the ability to (1) infect macrophages and epithelial cells, (2) produce distinct cytopathologic effect in alveolar and peritoneal macrophages and Vero cells, and (3) produce pocks on the chorioallantoic membrane of embryonated chicken eggs. Four vaccine strains and 5 field isolates were used in the study. The 5 field isolates were obtained directly from canine tissues. Of the 3 properties studied, only the comparison of the ability of the viruses to infect macrophages and epithelial cells was a consistent marker of virus origin. Virulent field isolates would only infect macrophage cultures, whereas the vaccine strains infected both types of cells. One avirulent field isolate from a case of old dog encephalitis reacted more like a vaccine strain by infecting both cell types.
Anamnestic Immune Response of Pigs to Pseudorabies Virus: Latent Virus Reactivation versus Direct Oronasal and Parenteral Exposure to Virus
Authors: Mengeling W. L.
Published: May 2011

The humoral antibody response of pseudorabies-immune pigs to reactivation of latent pseudorabies virus (PRV) was compared with the response following direct exposure to virulent PRV. Nine pigs that had been vaccinated for pseudorabies and later exposed to virulent virus to establish latent infection were given dexamethasone to reactivate latent virus (3 pigs), were exposed oronasally and parenterally to virulent virus (3 pigs), or were kept as nontreated controls (3 pigs). Sera collected from all 9 pigs just before and 3 weeks after treatment were tested by virus neutralization and radioimmunoprecipitation. The 3 pigs exposed directly to virulent virus and 2 of the 3 pigs given dexamethasone had a 4-fold or greater increase in neutralizing antibody titer. All 6 of these pigs had an increase in precipitating antibody activity. Precipitation patterns changed both quantitatively and qualitatively, especially for virus-coded proteins of relatively low molecular weight (<46 K). There were some differences in the precipitation patterns associated with sera of individual pigs. However, there was no clear indication of any difference between the 2 treatment groups and therefore no evidence that reactivation of latent virus is associated with any unique immunologic response that could be detected by radioimmunoprecipitation and used diagnostically. Clinical signs were limited to the 3 pigs that were exposed oronasally and parenterally to virulent virus even though the dexamethasone-treated pigs shed more virus for much longer than did those exposed directly to virus.
Experimental Equine Herpesvirus-l Infection in Llamas (Lama Glama)
Authors: House James A. , Gregg Douglas A. , Lubroth Juan , Dubovi Edward J. , Torres A.
Published: May 2011

Three llamas (Lama glama) were experimentally infected intranasally with an isolate of equine herpesvirus-1 (EHV-1) from the brain of an alpaca that had experienced severe neurologic signs. Two of the 3 llamas developed severe neurologic disorders following inoculation; 1 died, and 1 was euthanized in a moribund state. The third llama showed only mild neurologic signs. The euthanized llama had preexisting antibodies to-EHV-1, and the remaining 2 llamas were seronegative (virus neutralization titer < 6) at the time of inoculation. One of the seronegative llamas died acutely without production of detectable antibodies, and the other developed antibodies typical of a primary immune response. The EHV-1 virus was recovered only from a sample of the thalamus of the llama that died acutely. Histopathologic lesions were observed in the brain and retina of the dead and euthanized animals. This study verifies the pathogenic potential of EHV-1 for llamas.
A Competitive ELISA for Detection of Antibodies to the Group Antigen of Bluetongue Virus
Authors: Reddington J. J. , Reddington G. M. , MacLachlan N. J.
Published: May 2011

A competitive enzyme-linked immunosorbent assay (cELISA) was developed to detect antibodies to the group antigen of bluetongue virus (BTV). The epitope recognized by the BTV-specific monoclonal antibody was confirmed, by immunofluorescence staining of monolayers of virus-infected Vero cells, to be present on BTV serotypes 2, 10, 11, 13, and 17 but not on epizootic hemorrhagic disease virus (EHDV) serotypes 1 and 2. Sera from BTV-inoculated ruminants and rabbits were used to evaluate the cELISA and to compare its specificity and sensitivity with that of the conventional BTV-specific agar gel immunodiffusion (AGID) and serum neutralization (SN) tests. Rabbit antisera to the 5 serotypes of BTV present in the United States had cELISA titers (inverse of the final dilution of serum that gave > 20% inhibition) that ranged from 32 to > 1,024. Seroconversion of the 8 calves and lambs inoculated with BTV was detected by all 3 serologic tests (SN, AGID, cELISA) by 6 weeks after inoculation. Specificity of the cELISA test was confirmed with bovine sera that contained neutralizing antibodies to EHDV but not to the 5 serotypes of BTV present in the United States; these sera gave positive results by AGID test but were negative by cELISA. The sensitivity and specificity of the cELISA test was further confirmed by analysis of a panel of bovine test sera supplied by the National Veterinary Services Laboratories, indicating that the cELISA is a superior test for detection of BTV group-specific antibodies in sera from ruminants in the United States.
Molecular Diagnosis of Alcelaphine Herpesvirus (Malignant Catarrhal Fever) Infections by Nested Amplification of Viral DNA in Bovine Blood Buffy Coat Specimens
Authors: Katz Jonathan , Seal Bruce , Ridpath Julia
Published: May 2011

A fragment of alcelaphine herpesvirus-1 (AHV-1; malignant catarrhal fever) DNA was subcloned into pUC 18 and sequenced. The subclone hybridized strongly to AHV-1 DNA, weakly to alcelaphine herpesvirus-2 (AHV-2) DNA, and not at all to DNA from bovine herpesvirus-1 (BHV-1; infectious bovine rhinotracheitis [IBR] virus), bovine herpesvirus-2 (BHV-2; bovine herpes mamillitis [BHM] virus), and bovine herpesvirus-4 (BHV-4; isolate DN599). A 2-stage (nested) polymerase chain reaction (PCR) diagnostic test was devised based on a portion of the subcloned AHV-1 DNA sequence. First and second stage amplified AHV-1 DNA targets were 487 and 172 base pairs (bp) in length, respectively. Unique Pvu II and Stu I restriction endonuclease cleavage sites confirmed the identity of amplified AHV-1 DNA. Five AHV-1 and 2 AHV-2 isolates were identically and specifically PCR positive. BHV-1, BHV-2, and BHV-4 viruses were negative by the same procedure. As little as 0.01 TCID50 AHV-1 was detected using the nested amplification procedure. Simple methods of buffy coat isolation from bovine blood were employed to prepare specimens for PCR. An AHV-1-infected calf was PCR positive from 3 to 77 days postinoculation (PI), with rising seroconversion first noted 14 days PI. The AHV-1 DNA sequence was 62% homologous to a portion of the Epstein-Barr virus genome. The nested PCR procedure may improve the viral diagnosis of clinical and subclinical alcelaphine herpesvirus infections.
Methods for Detection and Frequency of Contamination of Fetal Calf Serum with Bovine Viral Diarrhea Virus and Antibodies against Bovine Viral Diarrhea Virus
Authors: Bolin Steven R. , Matthews Peter J. , Ridpath Julia F.
Published: May 2011

Methods used by the National Animal Disease Center to test fetal calf serum for contamination with bovine viral diarrhea virus (BVDV) and antibodies against BVDV are described. Using those methods, virus was isolated from 332 of 1,608 (20.6%) lots of raw fetal calf serum obtained specifically for the Center and 93 of 190 (49%) lots of commercially available fetal calf serum. Virus neutralization and immunoperoxidase staining tests were used to detect antibodies against BVDV in 224 of the 1,608 (13.9%) lots of raw fetal calf serum. Both BVDV and antibodies against BVDV were detected in 50 lots of raw serum. The molecular specificity of antibodies against BVDV was determined by radioimmunoprecipitation. Lots of fetal calf serum that contained BVDV-specific antibodies that did not neutralize virus were identified.
Suitability of the Broth-Disk Elution Test for Evaluating Susceptibility of Obligate Anaerobes to Trimethoprim-Sulfonamides
Authors: Indiveri Mary C. , Hirsh Dwight C.
Published: May 2011

Species of anaerobic bacteria isolated from clinical veterinary specimens were tested for susceptibility to trimethoprim-sulfonamides by the broth-disk elution technique. Three different media were used for each organism: prereduced anaerobically sterilized (PRAS) brain-heart infusion broth (BHI), thioglycollate broth, and a semidefined PRAS medium. Susceptibility results from these media were compared with those determined by interpreting the minimal inhibitory concentration obtained using an agar dilution technique. Results from broth-disk testing in semidefined medium agreed in 68.7% of the cases, in 53.7% for thioglycollate broth, and in 36.9% for BHI. The greatest deviation between techniques occurred with isolates belonging to the genus Bacteroides, followed by those of the genus Clostridium and those of the genus Fusobacterium. This deviation was directly proportional to increasing concentrations of thymidine in the BHI and thioglycollate broths but not with the semidefined medium. We conclude that the broth-disk elution method for measuring susceptibility of obligate anaerobes to trimethoprim-sulfonamides is unsuitable.
Use of a Salmonella Typhimurium-Derived Virulence Probe in the Detection of Salmonella sp. and in the Characterization of S. Cholerae-suis Virulence Plasmids
Authors: Maddox Carol W. , Fales William H.
Published: May 2011

A genetic probe encoding a virulence gene from Salmonella typhimurium was useful in the detection of Salmonella from feces during an outbreak of salmonellosis at a local dairy. A 3.2-kb BamHI restriction endonuclease fragment of the S. typhimurium virulence plasmid, pStSR100, has been useful as a DNA probe for both detection of Salmonella sp. and characterization of virulence plasmids from numerous field isolates. This virA probe hybridizes to a highly conserved gene carried on the large virulence plasmids of invasive Salmonella isolates. Colony blots prepared from feces directly plated onto MaConkey's agar failed to detect low numbers of Salmonella sp. However, hybridization of the virA probe to vacuum blots or colony blots prepared from feces in tetrathionate enrichment broth incubated for 16 hours at 37 C was effective for detecting Salmonella sp. and resulted in an 85.9% correlation with culture results. The probe also demonstrated the highly conserved nature (96%) of the virulence gene among S. cholerae-suis isolate plasmids detected using Southern blot analysis.
An Outbreak of Equine Neonatal Salmonellosis
Authors: Walker R. L. , Madigan J. E. , Hird D. W. , Case J. T. , Villanueva M. R. , Bogenrief D. S.
Published: May 2011

An outbreak of salmonellosis in foals occurred on a large Thoroughbred farm in California. Only foals <8 days of age exhibited clinical signs, which included depression, anorexia, and diarrhea. Three foals died from septicemia. The agent responsible was Salmonella ohio, which is rarely involved in salmonellosis in horses. During the course of the outbreak, S. ohio was isolated from 27 of 97 mares (27.8%) and 34 of 97 foals (35.1%). Mares were the presumed source of infection for foals. The absence of clinical signs in mares allowed for increased exposure of foals through environmental contamination. Although foals continued to become infected after strict control measures were adopted, none became ill. Salmonella serotypes of seemingly low virulence can produce serious disease outbreaks.
Immunocytochemical Detection of Ehrlichia Platys Antigens in Canine Blood Platelets
Authors: Simpson R. Mark , Gaunt Stephen D.
Published: May 2011

An avidin-biotin immunoperoxidase complex (ABC) immunocytochemical (ICC) stain procedure was optimized for detection of Ehrlichia platys antigens. Positive immunoreactivity was detected with dilutions of canine immune serum on acetone-fixed smears of platelet-rich plasma from E. platys-infected dogs. No E. platys antigens were detected when this ICC stain was applied to frozen or paraffin-embedded formalin- or acetone-fixed tissue sections from dogs with acute E. platys infection. Acetone fixation and freezing preserved ICC staining of ehrlichial antigens in infected blood platelets, whereas formalin treatment of similarly preserved E. platys -infected platelets nullified positive immunoreactivity. Significant E. platys infection of cells and tissues other than platelets may not occur.
Evaluation of Serologic Methods for the Detection of Antibodies to Encephalomyocarditis Virus in Swine Fetal Thoracic Fluids
Authors: Kim Hyun S. , Joo Han S. , Christianson William T. , Morrison Robert B.
Published: May 2011

Hemagglutination inhibition (HI) and agar gel immunodiffusion (AGID) tests were compared to the serum neutralization (SN) test to evaluate their ability to detect antibodies to encephalomyocarditis virus (EMCV). Swine fetal thoracic fluids of known EMCV SN antibody titers (200 samples 2 1:2, 100 samples < 1:2) were selected from a collection of field cases. The thoracic fluids were tested for EMCV antibodies by HI and AGID, and the results were compared to those of the SN test. Of 200 SN antibody-positive samples, 183 (9 1.5%) and 173 (86.5%) were positive in HI and AGID tests, respectively. Of 100 SN-negative samples, 81 (8 1%) and 94 (94%) were negative in HI and AGID tests, respectively. Agreement between the tests was analyzed by calculating Kappa values. The values were 0.73 between SN and HI tests and 0.77 between SN and AGID tests, indicating very good to excellent agreement for HI and AGID tests with the SN test. Of 200 SN-positive samples, 19 samples with low SN titers (1:2–1:16) were further tested by Western immunoblotting, and all were confirmed as positive. Interpretation of the present results suggests that both HI and AGID tests can be used as alternatives to the SN test.
Serologic Evidence of Coronavirus Infection in New York and New England Dairy Cattle with Winter Dysentery
Authors: Van Kruiningen H. J. , Castellano V. P. , Torres A. , Sharpee R. L.
Published: May 2011

Acute and convalescent sera were collected from 8 dairy herds with classic clinical features of winter dysentery. An enzyme-linked immunosorbent assay was used to measure coronavirus antibody titers, employing calf diarrhea coronavirus as antigen. Twenty-two of the 35 animals tested (63%) showed a ≥4-fold seroconversion. Adult cattle in all 8 herds seroconverted. These findings complement previously reported immunoperoxidase and electron microscopic evidence, suggesting an etiologic role for an enteric coronavirus in this disease.