bluuebunny/crossref_metadata_2025 huggingface dataset:
An Inherited Lower Motor Neuron Disease of Pigs: Clinical Signs in Two Litters and Pathology of an Affected Pig
Authors: O'Toole Donal , Ingram James , Welch Val , Bardsley Katie , Haven Tom , Nunamaker Carol , Wells Gerald
Published: May 2011

A chronic progressive neurodegeneration, called hereditary porcine neuronal system degeneration (HPNSD), was recognized in a swine herd in Devon, England. Adult pigs that were presumed carriers of the dominantly inherited trait for HPNSD were transferred from England, where a breeding colony was maintained for 9 years, to the Wyoming State Veterinary Laboratory (WSVL) for study. Two litters of affected piglets were born to 2 carrier sows at the WSVL. Clinical signs of muscular tremors, paresis, or ataxia developed at 12–59 days of age in 4 of 6 liveborn pigs. Three other pigs were stillborn. In the 4 affected livebom pigs, clinical signs progressed and included symmetrical (3 pigs) or asymmetrical (1 pig) posterior paresis, bilateral knuckling of metatarsal-phalangeal or carpal joints, poor exercise tolerance, and in 1 pig, marked hind limb hypermetria. A 34-kg gilt exhibiting clinical signs of muscular tremors and posterior paresis and clinical signs for 22 days was euthanized and examined postmortem at 83 days of age. Apart from decubitus ulcers, gross lesions were absent. Microscopically, perikaryal vacuolation and osmiophilic lipid droplets were observed in atrophic alpha motor neurons in the spinal cord. There was axonal (Wallerian) degeneration in sulcomarginal and dorsal spinocerebellar tracts. Axonal degeneration also involved ventral but not dorsal spinal nerve roots, and was present in eight peripheral nerves sampled for histopathology. Changes in skeletal muscles were consistent with denervation atrophy and were most pronounced in M. tibialis cranialis of the 6 muscles sampled. Immunohistochemical staining of spinal cord for phosphorylated and nonphosphorylated neurofilaments did not reveal abnormal patterns, unlike some well-characterized inherited motor neuron diseases in other species.
Immunohistochemistry of Parasitic Subepidermal Vesiculobullous Disease in American Badgers (Taxidea Taxus)
Authors: O'Toole Donal , Welch Val , Williams Beth
Published: May 2011

Some populations of free-ranging American badgers (Taxidea taxus) develop a distinctive seasonal dermatitis due to the subcutaneous filariid Filaria taxideae. Subepidermal vesicles that contain filarial ova develop in thinly haired skin of the inguinal area, proximal thigh, and ventral abdomen. The purpose of this study was to establish by immunohistochemistry whether basement membrane components colocalized with the roof or floor of vesicles and to confirm that filarial ova occur in intradermal vessels. Samples of skin with characteristic F. taxideae-induced subepidermal vesicles were collected from 10 adult male (n = 8) and female (n = 2) badgers. Samples were fixed in formalin for 1–4 days and processed routinely into paraffin wax. Immunohistochemical staining for basement membrane was attempted with anti-collagen IV antibodies (AM168-5M, AR079-5R, AB748) and antilaminin antibodies (MA078-5C, AR078-5R, L-9393). Optimal results in skin from badgers were obtained using a biotin-streptavidin technique and AR079-5R (anti-human collagen IV) and AR078-5R (anti-murine laminin). There was positive staining of the floor of vesicles in 5 of 6 badgers tested with antibodies to laminin and collagen IV. In 5/10 badgers, filarial ova and first stage F. taxideae larvae were found in dilated vascular channels of the upper dermis, and these vessels stained positively for factor VIII-related antigen. The results suggest that F. taxideae-induced subepidermal separation occurs consistently in the lamina lucida portion of the basal lamina and that filarial ova occur in dermal vessels.
Comparison of Two Hydrated Sodium Calcium Aluminosilicate Compounds to Experimentally Protect Growing Barrows from Aflatoxicosis
Authors: Harvey Roger B. , Kubena Leon F. , Elissalde Marcel H. , Corrier Donald E. , Phillips Timothy D.
Published: May 2011

Two formulations of hydrated sodium calcium aluminosilicate (HSCAS-1 and HSCAS-3), anti-caking agents for mixed feed, were added to the diets of growing barrows and were evaluated for their potential to diminish the clinical signs of aflatoxicosis. The experimental design consisted of 8 barrows (2 replicates of 4 each/treatment) assigned to 1 of the following 6 treatment diets (total of 48): 1) 0 g of HSCAS-1 or HSCAS-3 and 0 mg of aflatoxin (AF)/kg of feed (control); 2) 5 g HSCAS-1/kg of feed; 3) 5 g HSCAS-3/kg of feed; 4) 3 mg AF/kg of feed; 5) 3 mg AF plus 5 g HSCAS-1/kg of feed; or 6) 3 mg AF plus 5 g HSCAS-3/kg of feed. Barrows were maintained in indoor concrete-floored pens, with feed and water -available for 28 days (from 8 to 12 weeks of age). Barrows were observed twice daily and weighed weekly, and blood samples were collected at day 28 for hematologic, immunologic, and serum biochemical measurements. At the termination of the study, barrows were euthanized and necropsied. Barrow body weight gains were diminished, compared to those of controls, by consumption of AF alone and both of the AF plus HSCAS diets; however, the AF plus HSCAS-1 and AF plus HSCAS-3 barrow body weight gains were significantly greater ( P < 0.05) than those of the AF-alone barrows, No toxic responses or performance differences were noticed for barrows consuming either of the HSCAS-alone diets. Serum concentrations of alkaline phosphatase, gamma glutamyltransferase, calcium, cholesterol, albumin, triglycerides, and urea nitrogen were altered in barrows of the AF-alone treatment. The use of HSCAS prevented most but not all of the AF-induced changes in biochemical values. Immunologic measurements that were adversely affected by AF included mitogen-induced lymphoblastogenesis and peritoneal macrophage activity and function. The addition of HSCAS to AF-contaminated diets protected barrows from some of these toxic changes. Although immunologic measurements in the AF plus HSCAS groups were significantly different than those of the AF-alone group, values were still not equivalent to those of controls. These findings suggest that HSCAS-1 and HSCAS-3 are equal in their ability to protect against the toxicity of AF. Although these compounds may offer a novel approach to the preventive management of aflatoxicosis in livestock and poultry, HSCAS is not approved by the US Food and Drug Administration for treatment of animal diets for prevention of mycotoxicosis.
Immunoperoxidase Test for Rabies: Utility as a Diagnostic Test
Authors: Hamir A. N. , Moser G.
Published: May 2011

Although a fast and reliable fluorescent antibody (FA) test for rabies diagnosis is available, ideally the diagnostic procedure requires fresh or frozen brain tissues. In some instances (and particularly for retrospective studies), fresh or frozen tissues may not be available. In such cases, immunohistochemical tests may be utilized. Because such tests have been used only in a limited number of cases, their diagnostic reliability has not been thoroughly evaluated. This study documents the results of a streptavidin-biotin complex (ABC) immunoperoxidase test on formalin-fixed, paraffin-embedded brain tissues of domestic and wild animals that were positive on FA test for rabies prior to fixation. Thirty-nine of 40 rabies cases were positive with the ABC technique. Based on these preliminary results, the ABC technique appears reliable. However, more corroborative test results are needed before the ABC method can be used as a routine diagnostic procedure, especially with field samples and negative controls.
A Survey of G6 and G10 Serotypes of Group a Bovine Rotaviruses from Diarrheic Beef and Dairy Calves using Monoclonal Antibodies in ELISA
Authors: Lucchelli A. , Kang S. Y. , Jayasekera M. K. , Parwani A. V. , Zeman D. H. , Saif L. J.
Published: May 2011

Group A bovine rotaviruses (BRV) have been identified worldwide as a major cause of diarrhea in the young of many species, including humans. Group A rotaviruses are classified into serotypes on the basis of the outer capsid proteins, VP7 (G types) and VP4 (P types). To date, there are 14 G types of group A rotaviruses, with G1, G6, G8, and G10 described for BRV isolates. In this study, G6- and G lo-specific monoclonal antibodies (MAbs) were used in an enzyme-linked immunosorbent assay (ELISA) for the G typing of BRV-positive stool samples from diarrheic beef and dairy calves from South Dakota, Ohio, Michigan, Nebraska, and Washington, USA, and Ontario, Canada. ELISA plates were coated using a broadly reactive VP7 MAb (Common 60) or with G6- or G10-specific MAbs. BRV-positive fecal samples were diluted and added to duplicate wells, followed by the addition of polyclonal guinea pig anti-group A rotavirus serum as the secondary antibody. Several reference G6 and G10 BRV strains as well as other G types previously reported in cattle (G1, G2, G3, G8) and BRV-negative samples were included as G type specificity and negative controls. From a total of 308 field samples analyzed, 79% (244/308) tested positive by the broadly reactive VP7 MAb; of these, 54% (131/244) were G6 positive, 14% (35/244) were G10 positive, 4% (9/244) were both G6 and G10 positive, and 28% (69/244) were G6 and G10 negative. The negative samples may represent additional or undefined serotypes. The 89 samples from South Dakota were further subdivided into samples from beef ( n = 43) or dairy ( n = 46) herds. G6 was more prevalent in beef herd samples (67%) than in dairy herd samples (47.5%). In addition, dairy herds had higher percentages of G10-positive samples (17.5%) G6-G10 double positives (10%), and untypable samples (25%) than did beef herds, in which the prevalence of G10 positive samples was 5.5%, G6-G10 double positives was 5.5%, and untypable samples was 22%. Application of the serotype ELISA for the analysis of additional BRV samples will provide further epidemiologic data on the distribution of BRV serotypes in beef or dairy cattle, an important consideration for the development of improved BRV vaccines.
Comparison of Particle Concentration Fluorescence Immunoassay to Card and Complement Fixation Tests Using Isolation of Brucella Abortus as the Standard
Authors: Greenlee Michael T. , Farrar Jeff A. , Hird David W. , Holmes John C.
Published: May 2011

Serologic test data and bacteriologic culture results from 816 cattle were evaluated. Brucella abortus (field strain or strain 19) was isolated from 27.3% of the cows. Results of the card test, particle concentration fluorescence immunoassay (PCFIA), and complement fixation (CF) test were compared. Antibody titers were directly associated with B. abortus isolation, with the majority (64.7%) of field strain isolations having a PCFIA value of ≤0.300 and a CF test result of at least 1 + at a 1:160 dilution. The specificity of the CF test was significantly higher than that of the PCFIA (at CF ≥ 1:40 and PCFIA ≥ 0.300).
A Sensitive Immunoblotting Technique for the Serodiagnosis of Brucella Ovis Infections
Authors: Kittelberger Reinhold , Hansen Mike , Ross Gail P. , Hilbink Frans
Published: May 2011

A simplified electrophoretic immunoblotting technique based on antigen extracted from Brucella ovis cells with sodium dodecyl sulfate/mercaptoethanol was compared with the complement fixation test (CFT), the enzyme-linked immunosorbent assay, and the gel diffusion test. Sera from 89 chronically infected, semen culture-positive rams, 378 sera from B. ovis-infected flocks, 300 sera from accredited disease-free flocks, and 29 sera from specific-pathogen-free sheep were used. The immunoblotting technique had sensitivity and specificity comparable to those of the standard tests and was able to identify several CFT-negative or -borderline sera as positive. The major immunoreactive antigens of B. ovis had molecular masses of 63,29, 19 kD (proteins) and 8-12 kD (rough lipopolysaccharide). Antibodies against these antigens were present in 96% of CFT-positive sera from infected flocks and in 100% of sera from semen culture-positive rams. However, immunoblotting also identified antibodies to components other than the major antigens in 1% of CFT-negative sera from infected flocks and in 7.7% of the sera from flocks with a history of freedom from the disease. These reactions probably represent cross-reactivities with other microorganisms and were distinguishable from truly positive reactions.
Seroprevalence of Bovine Anaplasmosis in Oklahoma from 1977 to 1991
Authors: Rodgers S. J. , Welsh R. D. , Stebbins M. E.
Published: May 2011

The prevalence of anaplasmosis in Oklahoma cattle was determined on the basis of the standardized Anaplasma marginale complement fixation test on 20,155 sera submitted to the Oklahoma Animal Disease Diagnostic Laboratory during a 15-year period. Rates of seropositivity ranged from 4.7% to 17.6% on samples submitted for anaplasmosis testing of adult cows. The geographic distribution of recorded cases of anaplasmosis was 35 Oklahoma counties in 1977 and 48 Oklahoma counties in 1991.
Experimental Toxoplasmosis in Japanese Quail
Authors: Dubey J. P. , Goodwin M. A. , Ruff M. D. , Kwok O. C. H. , Shen S. K. , Wilkins G. C. , Thulliez P.
Published: May 2011

Twenty-four 5-month-old battery-hatched Japanese quail were inoculated orally with 105 (ME 49 strain, group A, 6 birds), 103 (ME 49 strain, group B, 6 birds), 105 (GT-1 strain, group C, 6 birds), and 103 (GT-1 strain, group D, 6 birds) Toxoplasma gondii oocysts. All birds in group C died or were euthanized within 8 days after inoculation (DAI). Five of the 6 birds in group D died or were euthanized 8, 9, 15, 19, and 23 DAI. One of the 6 quail in group A died 9 DAI, and 1 of the 6 birds in group D died 16 DAI. The 11 quail (1 from group D and 10 from groups A and B) were euthanized 63 DAI; T. gondii was isolated by bioassays in mice from the brains of 10, hearts of 10, and skeletal muscles of all 11 quail. Quail that survived marked small intestinal and splenic toxoplasmosis lived long enough to develop severe protozoal pneumonia, myocarditis, or meningoencephalitis. The quail that survived only to be examined at 63 DAI had moderate multifocal nonpurulent encephalitis and myositis and had a hypertrophic spleen that contained hemosiderin-laden macrophages. Toxoplasma gondii antibodies were found in the sera of all quail examined 63 DAI. Antibody titers to T. gondii in the modified agglutination test were higher than in the indirect hemagglutination and latex agglutination tests. Antibodies were not detected in quail sera examined by the Sabin-Feldman dye test.
Antigens of Adults and Third-Stage Larvae of the Meningeal Worm, Parelaphostrongylus Tenuis (Nematoda, Metastrongyloidea)
Authors: Neumann N. F. , Pon W. S. , Nowicki A. , Samuel W. M. , Belosevic M.
Published: May 2011

We studied the antigens of adults and third-stage larvae of the meningeal worm, Parelaphostrongylus tenuis, in an attempt to identify potential serodiagnostic molecules for this important infection of wild ungulates. Soluble extracts of P. tenuis adult worms and third-stage larvae were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional (2D) electrophoresis and analyzed by immunoblotting using purified rabbit anti- P. tenuis immunoglobulin G (IgG). The IgG antibodies were obtained from animals immunized with P. tenuis adult worm or third-stage larva soluble extract and serum from elk infected with P. tenuis. Out of more than 75 antigens (as shown by 2D electrophoresis and immunoblotting), 7 antigens from adults (four 170-120-kD molecules with isoelectric points between 6.0 and 6.6, two 55kD molecules with isoelectric points of 5.6 and 5.8, and one 13-kD molecule) and 2 antigens from third-stage larvae (one 25-30-kD molecule with an isoelectric point of 6.3 and one 13-kD molecule) distinguished P. tenuis from two other nematodes, Dictyocaulus viviparus and Trichinella spiralis. Initial results using serum from experimentally infected elk indicate that this serum recognized a similar profile of P. tenuis antigens when compared with the serum from immunized rabbits. This research has set the foundation for the development of a test for P. tenuis infections in wild and recently domesticated elk and other ungulates.
Effect of Time and Storage Temperature on Cholinesterase Activity in Blood from Normal and Organophosphorus Insecticide-Treated Horses
Authors: Plumlee Konstanze H. , Richardson Elizabeth R. , Gardner Ian A. , Galey Francis D.
Published: May 2011

Delays between time of sampling and time of testing are common; therefore, the length of time that blood can be stored at various temperatures was evaluated for effects on cholinesterase activity. Six horses were treated with 16 g of trichlorfon per os, 6 horses were treated with 15 g of dichlorvos per os, and 10 horses were untreated controls. The cholinesterase activity in whole blood from each horse was measured using an adaptation of the Ellman calorimetric method. The blood from each horse was then divided into 3 groups and stored at 5 C (refrigerated), 20 C (room temperature), or 38 C (incubated). Subsequent cholinesterase activities were measured daily and then at weekly intervals. The cholinesterase activities did not significantly increase or decrease ( P > 0.05) in the blood from the untreated horses until after 1 week for any of the 3 temperature groups. The cholinesterase activities did not significantly increase or decrease ( P > 0.05) in the stored blood from the trichlorfon-treated horses for 4 weeks in all 3 temperature groups. The cholinesterase activities significantly increased ( P < 0.05) in the stored blood from the dichlorvos-treated horses after 1 week when the blood was refrigerated and by 24 hours when the blood was stored at room temperature or incubated. Therefore, blood from normal or organophosphate-treated horses can be used for cholinesterase evaluation for up to 1 week when stored at 5 C.
Canine Distemper Epizootic in Lions, Tigers, and Leopards in North America
Authors: Appel Max J. G. , Yates Rebecca A. , Foley George L. , Bernstein Jon J. , Santinelli Silvio , Spelman Lucy H. , Miller Lyle D. , Arp Lawrence H. , Anderson Mark , Barr Margaret , Pearce-Kelling Susan , Summers Brian A.
Published: May 2011

Canine distemper virus (CDV) infection occurred in captive leopards ( Panthera pardus), tigers ( Panthera tigris), lions ( Panthera leo), and a jaguar ( Panthera onca) in 1991 and 1992. An epizootic affected all 4 types of cats at the Wildlife Waystation, San Fernando, California, with 17 mortalities. CDV-infected raccoons were thought to be the source of infection in these cats. Two black leopards died at the Naibi Zoo, Coal Valley, Illinois, and 2 tigers died at the Shambala Preserve, Acton, California. Initial clinical signs were anorexia with gastrointestinal and/or respiratory disease followed by seizures. Canine distemper virus was isolated from 3 leopards, 3 tigers, and 3 lions that died or were euthanized when moribund. Monoclonal antibody testing identified the virus isolates as CDV. Gross and histopathologic findings were similar to those found in canids with distemper with a few exceptions. There were fewer lesions in the brain, and there was a pronounced type 2 cell proliferation in the lung, with inclusion bodies and CDV antigen demonstrated by immunohistology. Neutralizing antibody to CDV was found in high titers in serum from most animals but was absent or was found only in low titers in some cats that succumbed after CDV infection. There was a marked difference in neutralizing antibody titers when tests were done with different strains of CDV.
Cloning, Expression, and Sequence Analysis of the ORF4 Gene of the Porcine Reproductive and Respiratory Syndrome Virus MN-1b
Authors: Kwang Jimmy , Kim Hyun Soo , Joo Han S.
Published: May 2011

Porcine reproductive and respiratory syndrome virus (PRRSV) MN-1b strain open reading frame 4 (ORF4) has been cloned, sequenced, and expressed in Escherichia coli. The homologies of nucleotide and amino acid sequences between MN-1b (US isolate) and LV (European isolate) are 69% and 64%, respectively. The data also showed that ORF4 of MN-1b is 36 bases shorter than that of LV. Western blot analysis of expressed recombinant ORF4 protein reacted with 65% (26/40) of PRRSV-infected pig sera tested. These results demonstrated that ORF4 of PRRSV may not be a well-conserved region.
Importance of Primer Selection in the Application of PCR Technology to the Diagnosis of Bovine Leukemia Virus
Authors: Marsolais Grégoire , Dubuc Roger , Bergeron Jean , Morrey John D. , Kelly Emma J. , Jackson M. Keven
Published: May 2011

The polymerase chain reaction (PCR) was used to detect bovine leukemia virus in bovine blood samples. When applied to leucocytes extracted from the blood samples, the standard method of DNA extraction gave good correlation with agar gel immunodiffusion, but a method in which 5 μl of blood was the starting material was unreliable. Selection of the primers was important, and differences in results were observed when the PCR method was applied to blood samples from different geographic areas. The sensitivity varied from 50% to 90%, depending on the primer set applied to the gag gene of proviral nucleic acid. This variation was based on geographic origin of the cattle, suggesting an influence of viral strain. In some areas, more than 1 primer may needed to optimize results.
Detection of Bovine Group B Rotaviruses in Feces by Polymerase Chain Reaction
Authors: Chinsangaram Jarasvech , Akita Geoffrey Y. , Osburn Bennie I.
Published: May 2011

A pair of primers designed from the sequence of genome segment 9 of group B rat rotavirus (IDIR) were employed to amplify genome segment 9 of a group B bovine rotavirus in a polymerase chain reaction (PCR) and to sequence the derived PCR products. A new pair of primers were synthesized from the obtained sequence data and used in a PCR detection assay for group B bovine rotavirus in fecal samples. In addition, another pair of primers were designed to produce a PCR-derived internal probe. This probe was used in a chemiluminescent hybridization to confirm the specificity and to increase the sensitivity of the assay. This assay could detect 0.1 fg of target double-stranded RNA. It was specific to group B bovine rotavirus and did not detect group B rat (IDIR) and porcine rotaviruses, group A bovine (NCDV), simian (SA-11), equine (H-2), porcine (OSU), human (DS-1), deer, and avian rotaviruses, coronavirus, or other enteric organisms tested in this study.
An Indirect Microimmunofluorescence Test for Detection of Chlamydial Antibodies in Ovine Fetal Fluids
Authors: Sanderson T. P. , Andersen A. A. , Miller L. D. , Andrew J. J. , Janke B. H. , Larson D. L. , Schwartz K. J.
Published: May 2011

The objective of this study was to evaluate an indirect microimmunofluorescence test (IMIF) for detection of Chlamydial antibodies in serum and/or thoracic fluids of aborted ovine fetuses. One hundred eighty-two ovine fetuses, including 64 fetuses from 40 ewes that were experimentally infected with an ovine abortion strain of Chlamydia psittaci at gestation days 90–100, 10 fetuses from 6 normal ewes, and 108 fetuses selected from those received at the Iowa Veterinary Diagnostic Laboratory, were evaluated in this study. Fetuses from experimentally infected ewes were examined 4–60 days after inoculation. The IMIF findings were compared with the results of complement fixation serology for Chlamydiae and concentrations of immunoglobulin (IgG). Chlamydiae-specific antibodies were detected by IMIF in 28 of 38 fetuses infected with C. psittaci. Elevated levels of IgG and IMIF titers ≥ 1:8 were consistent findings in ovine fetuses infected with Chlamydiae for more than 24 days. IgG levels and titers of Chlamydial antibodies increased with maturity of the fetus and duration of Chlamydial infection. Chlamydial antibodies were not detected with the complement fixation test. Fluids from ovine fetuses aborted as a result of other causes also were examined, and IMIF results were negative. The results of this study indicate that the IMIF is a useful and relatively rapid test for identification of Chlamydial antibodies in ovine fetuses.
Streptococclcs Suis Infection in Swine: A Retrospective Study of 256 Cases. Part II. Clinical Signs, Gross and Microscopic Lesions, and Coexisting Microorganisms
Authors: Reams Rachel Y. , Glickman Lawrence T. , Harrington Daniel D. , Thacker H. Leon , Bowersock Terry L.
Published: May 2011

A retrospective study of 256 cases of naturally acquired Streptococcus suis infections in swine submitted to the Indiana Animal Disease Diagnostic Laboratory from 1985 to 1989 was undertaken to describe the clinical signs, lesions, and coexisting organisms associated with S. suis serotypes 1–8 and 1/2. Infected pigs generally had clinical signs and gross lesions referable to either the respiratory system or to the central nervous system (CNS), but not both. Neurologic signs were inversely related to gross lesions in the respiratory tract ( R2 = −0.19, P = 0.003), as were respiratory signs and gross lesions in the CNS ( R2 = −0.19, P = 0.003). Suppurative bronchopneumonia was the most common gross lesion observed (55.2%, overall). Fibrinous and/or suppurative pleuritis, epicarditis, pericarditis, arthritis, peritonitis, and polyserositis were also reported. In 68% of the pigs, other bacteria in addition to S. suis were isolated. Escherichia coli (35.0%) and Pasteurella multocida (30.0%) were the most commonly recovered bacterial agents. Mycoplasma and viral agents were identified less often, and their role in the development of streptococcosis was difficult to assess. In pigs infected with serotypes 2–5, 7, 8, and 1/2, suppurative meningitis with suppurative or nonsuppurative encephalitis, suppurative bronchopneumonia, fibrinopurulent epicarditis, multifocal myocarditis, and cardiac vasculitis were the most common microscopic lesions observed, whereas pigs infected with serotype 1 generally presented with suppurative meningitis and interstitial pneumonia. Microscopic lesions were morphologically similar among serotypes and were also similar to those reported with other pyogenic bacteria. The distribution of clinical signs and the gross and microscopic lesions in pigs infected with S. suis varied among serotypes. However, these differences were not statistically significant and could not be used to distinguish between the various serotypes. These findings suggest that in pigs infected with S. suis, suppurative or fibrinopurulent inflammation in brain, heart, lungs, and serosae predominates and that bacterial culture is needed to confirm a diagnosis of streptococcosis in swine and to differentiate this disease from those caused by other pyogenic bacteria.
Mycoplasma Infection in a Commercial Goat Dairy Caused by Mycoplasma Agalactiae and Mycoplasma Mycoides Subsp. Mycoides (Caprine Biotype)
Authors: Kinde Hailu , DaMassa Al J. , Wakenell Patricia S. , Petty R.
Published: May 2011

A commercial dairy goat herd of 600 animals experienced sudden onset of arthritis/polyarthritis, clinical mastitis, and sudden death in does. The offending infectious agents were Mycoplasma agalactiae and M. mycoides subsp. mycoides (caprine biotype). The disease syndrome began approximately 4 weeks following the 1) introduction into the herd of a lactating doe with no apparent clinical signs and 2) a breakdown of proper hygienic conditions in the milking parlor. Over a period of 3 weeks, 90 does (15%) either died or were culled because of arthritis/polyarthritis and mastitis. A management decision resulted in only the does affected with M. mycoides subsp. mycoides being submitted for necropsy; those affected with M. agalactiae, which were in a different “string,” were not submitted for evaluation. Gross necropsy of the does affected with M. mycoides subsp. mycoides showed purulent discharges from the udders, enlarged supramammary lymph nodes, enlarged and firm spleens, and swollen livers. Microscopic findings were characterized by a loss of vascular integrity and diffuse fluid leakage in multiple organs. Antibiotic therapy with tylosin was attempted but was not successful. The outbreak was terminated following the removal or segregation of affected does and implementation of hygienic conditions in the milking parlor.
Molecular and Biological Characterization of a Newly Isolated Anaplasma Marginale Strain
Authors: Eriks Inge S. , Stiller David , Goff Will L. , Panton Marc , Parish Steven M. , McElwain Terry F. , Palmer Guy H.
Published: May 2011

Anaplasma marginale, a rickettsial hemoparasite of cattle and other ruminants, results in significant economic losses worldwide. Distinct strains of A. marginale have been identified based on differences in tick transmissibility, molecular size of surface proteins and DNA restriction fragments, and reactivity to a panel of monoclonal antibodies. These different strains vary considerably in their virulence, antigenic composition, and ability to protect against heterologous challenge. In this paper, we report on the molecular characterization of a newly isolated strain of A. marginale, designated St. Maries, recovered from an acutely infected cow in northern Idaho. Dermacentor andersoni ticks taken from the infected animal were tested for infection by RNA probe analysis. The infection rate of male ticks (as determined by midgut infection) was 100%, and the infection rate of female ticks was 83%. Infected male ticks were able to transmit the St. Maries strain to a susceptible calf. The high infection rate in male ticks may be particularly relevant, given that male ticks are believed to be epidemiologically important in transmission of A. marginale because of their intermittent feeding behavior, which promotes interhost transfer. The newly isolated strain differs from other US strains, including strains previously isolated in Idaho and Washington, based on reactivity to a panel of monoclonal antibodies and restriction fragment length polymorphisms. These results imply that antigenically distinct strains of A. marginale may arise within the same region. However, because epitopes previously demonstrated to be conserved in other A. marginale strains were also present in the St. Maries strain, it is possible to develop diagnostic tests against conserved epitopes that could be used for early identification of infected cows or ticks.
Canine Mast Cell Tumors: A Comparison of Staining Techniques
Authors: Simoes Jose Pedro Cannas , Schoning Polly
Published: May 2011

Twelve histochemical methods; affinity staining with avidin peroxidase, wheat germ agglutinin, and concavalin-A agglutinin; and an immunohistochemical stain with Kpl (CD68) antibody were compared for their relative effectiveness in staining canine mast cell tumors. Stains were compared in 28 mast cell tumors and 19 histiocytomas. The effectiveness of the histochemical methods and the lectins decreased as the mast cells became less differentiated. None of the staining methods were positive on histiocytomas. Periodic acid-Schiff (PAS) gave positive results in a few cases of mast cell tumors where other histochemical stains were negative. Although avidin peroxidase and Kpl antibody stained more mast cell tumors than any other method, they did not differ significantly from Luna's method, toluidine blue pH 0.5, toluidine blue pH 4.5, alcian blue pH 2.5, safranin O, Unna's method, and Giemsa. No stain was ideal for the diagnosis of canine mast cell tumors; however, this study suggests that the use of avidin peroxidase, Kpl antibody, and PAS may give additional information for individual poorly differentiated tumors without substantial increase in time or cost.
Diagnosis of Porcine Reproductive and Respiratory Syndrome
Authors: Mengeling William L. , Lager Kelly M. , Vorwald Ann C.
Published: May 2011

The most suitable tissue samples and test procedures for the etiologic diagnosis of porcine reproductive and respiratory syndrome (PRRS) were found to depend on several variables including the age of the pig from which tissues were collected, the stage of infection (acute or persistent), the available complement of diagnostic reagents, and the urgency in obtaining results. When the diagnosis involved acute infection of congenitally or neonatally infected pigs, and susceptible cell culture(s) was available for virus isolation, then both serum and alveolar macrophages (AM) were reliable samples. Alveolar macrophages flushed from infected lungs provided a temporal advantage, however, in that in addition to their use for virus isolation, i.e., from a lysate of AM, they could be cultured in vitro and examined for the presence of viral antigens by immunofluorescence microscopy (FA) as early as 1 hour after they were added to the culture vessel. The examination of AM in this manner also circumvented the need for additional cell cultures to test for infectious virus. Testing presuckhng sera by indirect FA for antibodies to PRRS virus also was of diagnostic value and, like FA with AM, could be completed soon after sample collection. For older pigs, AM were more reliable than serum, lungs, or any of 27 other tissues evaluated as diagnostic samples and were often the only samples in which infectious virus and viral antigens were detected when pigs were euthanized more than 3 weeks postexposure. A simple procedure for on-farm collection of AM as well as methods for testing AM for viral antigens and neonatal (presuckling) sera for homologous antibody in a modestly equipped laboratory, such as one that might be maintained by a veterinary practitioner, are described and discussed.
Early Infection of Interdigitating Dendritic Cells in the Pig Lymph Node with African Swine Fever Viruses of High and Low Virulence: Immunohistochemical and Ultrastructural Studies
Authors: Gregg Douglas A. , Mebus Charles A. , Schlafer Donald H.
Published: May 2011

The role of interdigitating dendritic cells (IDCs) in the early pathogenesis of African swine fever (ASF) was investigated using mandibular lymphoid tissue from normal pigs and pigs inoculated oronasally with highly virulent Lisbon 60 (L-60) and moderately virulent Dominican Republic 1979 (DR-2) ASF virus (ASFV) isolates. Paraffin-embedded tissue sections were immunostained for ASFV antigen and S-100 protein, a marker of IDCs, using an avidin-biotin alkaline phosphatase procedure. Swine IDCs were identified morphologically by light microscopy, electron microscopy, and S-100 immunostaining. Infection with ASFV caused a marked reduction in S-100 staining by 3 days postinfection (DPI) that persisted through 14 DPI. Early ASFV infection of IDCs was demonstrated at 3 DPI by double immunohistochemical staining of cryosections and by transmission electron microscopy. These results support the hypothesis that the failure of a humoral immune response to virulent ASFV may be due to a primary infection of IDCs and the inability of IDCs to initiate an immune response. Infection of IDCs has also been demonstrated with human immunodeficiency virus (HIV-1), and these infections have some aspects in common.
Detection and Subtyping of Foot-and-Mouth Disease Virus in Infected Cattle by Polymerase chain Reaction and Amplified VP1 Sequencing
Authors: Stram Yehuda , Molad Thea , Chai Dalia , Gelman Boris , Yadin Hagai
Published: May 2011

Fast and accurate detection of foot-and-mouth disease (FMD) outbreaks is needed to limit spread of the disease by proper vaccination. The use of the polymerase chain reaction (PCR) has revolutionized the way in which viral diseases are diagnosed. Sequence analysis of the amplified VP1 sequence can enable the classification of FMD virus detected in the morbid animal. PCR assays were carried out to identify the virus and its serotype in suspect animals from 2 outbreaks of FMD type O virus. Sequence analysis of the amplified VP1 cDNA showed 78% homology with O1K and over 95% homology between the samples. These findings suggest that the 2 outbreaks were due to infection with the same virus serosubtype.
Evaluation of an Antigen-Capture ELISA for Detection of Bovine Viral Diarrhea Virus in Cattle Blood Samples
Authors: Sandvik Torstein , Krogsrud Johan
Published: May 2011

From blood samples routinely received for bovine viral diarrhea (BVD) diagnosis, 860 samples without antibodies to BVD virus were examined in both virus isolation and in an antigen-detecting enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibodies (MAbs) against the nonstructural BVD virus protein p125/p80. A total of 843 samples (98%) were positive ( n= 170, 20%) or negative ( n = 673, 78%) in both tests, corresponding to an agreement of K = 0.94. Among 17 samples with diverging results, 3 were from animals transiently infected with BVD virus, and 5 came from clinically affected animals. The reactivity of the MAbs was controlled against 387 field isolates of BVD virus. All were detected by the MAbs, thereby confirming the general view that the p125 virus protein is highly conserved among different BVD viruses.
Immunoperoxidase Detection of Ophidian Paramyxovirus in Snake Lung using a Polyclonal Antibody
Authors: Homer Bruce L. , Sundberg John P. , Gaskin Jack M. , Schumacher Juergen , Jacobson Elliott R.
Published: May 2011

In a retrospective study of proliferative interstitial pneumonia in viperid and nonviperid snakes, formalin-fixed, paraffin-embedded lungs from 52 snakes were screened for immunohistochemical reactivity to ophidian paramyxovirus. All snakes were from zoological collections that experienced mortalities attributed to paramyxovirus infection. Of the 52 snakes, 47 had pulmonary lesions compatible with ophidian paramyxovirus infection. Histologic changes in affected lungs included hyperplasia and hypertrophy of septal and faveolar epithelial cells, loss of ciliated cells, mixed leukocytic interstitial infiltrates, fibrinonecrotic exudate in the lumen of proximal and distal faveolar compartments, and occasional epithelial syncytial cell formation or intraepithelial eosinophilic intracytoplasmic inclusions. Lungs were immunohistochemically stained for paramyxovirus antigens by utilizing rabbit polyclonal antibodies against a paramyxovirus isolate from a black mamba ( Dendroaspis polyepis polyepis). Virus infection in 6 snakes was confirmed by virus isolation from frozen lung tissue. Of the 6 lungs from which paramyxovirus was isolated, 5 lungs stained positively for viral antigens utilizing antisera to the black mamba isolate. Altogether, 36 lungs stained positively for paramyxovirus antigens. There was multifocal to diffuse linear staining of the lumenal surface of faveolar epithelium, and there were multiple foci of granular cytoplasmic staining. Immunohistochemical staining of formalin-fixed lungs from snakes with proliferative interstitial pneumonia was helpful as a routine diagnostic test for substantiating a diagnosis of ophidian paramyxovirus infection.
Outbreaks of Yersiniosis Caused by Yersinia Pseudotuberculosis in Farmed Cervids
Authors: Sanford S. Ernest
Published: May 2011

Yersiniosis, caused by Yersinia pseudotuberculosis, was diagnosed in 25 deer submitted to the Veterinary Diagnostic Laboratories of the Ontario Ministry of Agriculture and Food from 8 farms in Ontario, Canada, over the 4-year period of January 1990 to December 1993. The organism was cultured, usually in large numbers, from the intestines and, less frequently, mesenteric lymph nodes and/or the spleen of 13 red deer (Cervus elaphus), 6 elk (C. e. canadiensis), 4 fallow deer (Dama dama), and 2 red deer-elk hybrids. Outbreaks occurred almost exclusively in the fall (September-November) and affected recently weaned calves, 4–6 months old. Sudden death usually signalled the start of an outbreak. Outbreaks often occurred during inclement weather; usually a sudden cold snap, with or without rain, after a period of relatively mild fall weather. At necropsy, the animals were usually emaciated and dehydrated, and the perineum and tail were sometimes fecally stained. The small and large intestines were congested and filled with serofibrinous and/or seroanguineous fluid. Mesenteric lymph nodes were edematous and congested. Microabscesses surrounding large bacterial colonies in the lamina propria in the jejunum and ileum were typical lesions seen histologically. Peyer's patches were depleted and sometimes contained microabscesses. Purulent lymphadenitis associated with bacterial colonies was the prominent histologic lesion in mesenteric lymph nodes.
Detection of Anti-Listeriolysin O in Dairy Cattle Experimentally Infected with Listeria Monocytogenes
Authors: Baetz Albert L. , Wesley Irene V.
Published: May 2011

A dot-blot assay and an enzyme-linked immunosorbent assay (ELISA) to detect listeriosis in dairy cattle were developed that detected anti-listeriolysin O antibodies in the serum of cows experimentally infected with Listeria monocytogenes. The tests utilized purified listeriolysin O (LLO) as the detection antigen and streptolysin O (SLO) to absorb cross-reacting antibodies. The two tests were compared with an agglutination test that used formalin-killed whole L. monocytogenes cells. Blood samples were collected periodically from 17 cows after intramammary gland infection, and the development of anti-LLO antibodies was followed by an agglutination test, the dot-blot test, and the ELISA. In general, an agglutination titer of >640 was needed for a positive dot-blot anti-LLO test for nonpregnant cows. However, I pregnant cow with an agglutination titer of 20 was positive in the dot-blot test. The ELISA was as sensitive as the dot-blot assay but gave a quantitative measurement to distinguish serum samples of positive reactors from cross-reactors. The specificity of the LLO-based tests was further evaluated using serum from cows that had been experimentally infected with Staphylococcus aureus, 17 of which had agglutination titers for L. monocytogenes >640. These elevated agglutination titers were probably due to cross-reacting bacterial antigens because serum from 9 of 17 of these animals did not react to the purified LLO antigen. A positive response to the LLO-based dot-blot and ELISA assays is indicative of previous or current infection with L. monocytogenes.
Use of Commercial Enzyme Kits and Fatty Acid Production for the Identification of Serpulina Hyodysenteriae: A Potential Misdiagnosis
Authors: Milner Julie A. , Truelove Kirsty G. , Foster Richard J. , Sellwood Richard
Published: May 2011

The accuracy of identification of Serpulina hyodysenteriae strains grown in a complex medium was 90% when 2 commercial test kits were used. Unlike the other S. hyodysenteriae strains, S. hyodysenteriae strain P35/2 was unusual in being indole negative. The nonpathogenic intestinal spirochete PWS/A, which is from a different species, was indole positive and α-galactosidase negative. Identification of these spirochetes on the basis of these kits alone would have been incorrect. The analysis of volatile fatty acids by gas chromatography showed that the ratio of acetic to butyric acid was from 11:1 to 44:1 for S. hyodysenteriae strains, which distinguished them from the other spirochetes. The exception was PWS/A (acetic: butyric of 32:1), but this spirochete, unlike the S. hyodysenteriae spirochetes, also produced isobutyric acid. Short chain fatty acid (SCFA) analysis by high-performance liquid chromatography detected different SCFAs in addition to acetic and butyric acids. These additional SCFAs did not contribute to further differentiation of the porcine spirochetes.
Isolation and Culture of Large Vessel Endothelium from White-Tailed Deer (Odocoileus Virginianus)
Authors: Howerth Elizabeth W. , Stallknecht David E.
Published: May 2011

Endothelial cells were obtained from white-tailed deer carotid arteries and umbilical vessels by instilling a weak collagenase type II solution. Cell growth was best when cultures were grown in plates coated with either fibronectin or laminin. Several commercially available media supported the growth of these cells when supplemented with a commercially available endothelial growth supplement and 10% fetal calf serum. Cells obtained could be characterized as endothelium by ultrastructural characteristics and by the uptake of fluorescent acetylated low-density lipoprotein. Cells stained positive for factor VIII-related antigen up to passage 2, but staining was inconsistent by passage 3, and no immunoreactivity could be demonstrated after passage 4.
Evaluation and Application of an Indirect ELISA for the Detection of Antibodies to Bovine Respiratory Syncytial Virus in Milk, Bulk Milk, and Serum
Authors: Elvander M. , Edwards S. , Näslund IS. , Linde N.
Published: May 2011

An indirect enzyme-linked immunosorbent assay (ELISA) was developed at the National Veterinary Institute (NVI), Uppsala, to detect antibodies to bovine respiratory syncytial virus (BRSV) in serum and milk. For the evaluation of the NVI ELISA, field sera collected from cattle in England and Sweden were tested in parallel with an ELISA in use at the Central Veterinary Laboratory (CVL), Weybridge. The tests showed 96% agreement. The sensitivity and specificity of the NVI ELISA relative to the CVL ELISA were 94% and 100%, respectively. There was evidence that the difference in sensitivity between the 2 tests was due to the detection of both IgG and IgM class antibodies by the CVL ELISA, whereas the NVI ELISA was designed specifically to detect IgGl. Milk and serum samples from individual cows were tested by the NVI ELISA for presence of antibodies to BRSV. There was a good correlation between the ability to detect antibodies in serum and the ability to detect them in milk, although the antibody titer was generally lower in milk than in serum. Bulk milk samples were collected from farms with severe clinical symptoms of respiratory distress and from farms with no history of respiratory disease. There was a clear distinction between antibody levels in diseased and healthy herds. The NVI ELISA is a rapid and reliable test for detecting antibodies to BRSV in milk, bulk milk, and serum samples.
Development and Application of a Microneutralization ELISA for the Detection of Antibodies to Bovine Respiratory Syncytial Viruses
Authors: Ellis John A. , Hassard Lori , Morley Paul S.
Published: May 2011

A microneutralization enzyme-linked immunosorbent assay (ELISA) was developed to detect specific antibodies to bovine respiratory syncytial viruses (BRSVs) in cattle sera using a monoclonal antibody to the fusion protein of the virus. Serum from 20 naturally exposed, 24 experimentally infected, and 15 immunized cattle were evaluated using 3 different BRSV isolates. Antibody titers determined with the micro-neutralization ELISA were compared with those derived from a classical virus neutralization assay, an indirect ELISA, and a fusion inhibition assay. These studies demonstrated a high degree of correlation (usually 0.90) among the assays. Furthermore, the results showed that immunization of cattle with one isolate (subgroup) of BRSV induced antibody responses that cross-reacted with at least 2 disparate isolates. These results document the utility of the microneutralization ELISA in assessing functionally important antibody responses to BRSVs in cattle.