bluuebunny/crossref_metadata_2025 huggingface dataset:
A Retrospective Study on the Disappearance of Blood Lead in Cattle with Accidental Lead Toxicosis
Authors: Rumbeiha Wilson K. , Braselton W. Emmett , Donch Deborah
Published: May 2011

Lead poisoning in cattle and other food animals is of public health significance because of the potential for human exposure to lead through ingestion of contaminated meat and milk products derived from lead-poisoned animals. In Michigan, lead poisoning in livestock is a reportable disease, and positive cattle are quarantined until they test negative (<0.05 ppm blood lead). There is surprisingly little information on blood lead kinetics in cattle. The half-life has been variably reported as 9 weeks and 1–2 months. Because these data did not fit those obtained from cases received at the Michigan State University Animal Health Diagnostic Laboratory, a retrospective study was conducted to review all cases of accidental lead poisoning in cattle between 1990 and 1998. This information is needed to estimate when quarantined lead-poisoned cattle can be released. The results showed that the half-life of blood lead was quite variable and ranged from 48 to 2,507 days. The shortest half-lives (48, 56, and 57 days) were found in a lactating herd of 20-month-old heifers. The longest half-life, 2,507 days, was found in a 9-month-old castrated bull, which ingested a discarded automobile battery. Of the 24 animals monitored, only 8/24 (33%) had half-lives between 6 and 14 weeks. In conclusion, the half-life of blood lead is difficult to predict in accidental cases of cattle poisoning.
Seroconversion and Abortion in Cattle Experimentally Infected with Brucella sp. Isolated from a Pacific Harbor Seal (Phoca Vitulina Richardsi)
Authors: Rhyan Jack C. , Gidlewski Thomas , Ewalt Darla R. , Hennager Steven G. , Lambourne Dyanna M. , Olsen Steven C.
Published: May 2011

Previously unrecognized Brucella species have been isolated from a number of marine mammals, including harbor seals ( Phoca vitulina richardsi) in the Puget Sound area of the state of Washington. Because of the presence of dairy herds in proximity to the harbor seal populations, a study was conducted to determine the effects of the harbor seal Brucella isolate in experimentally inoculated cattle. Six pregnant cattle were exposed by intravenous injection ( n = 3) or intraconjunctival inoculation ( n = 3). Two pregnant cows were intravenously injected with saline and served as controls. All of the cows receiving the Brucella seroconverted on 1 or more tests commonly used for the detection of Brucella abortus infection. Two of the cattle receiving the intravenous inoculation aborted, and brucellae were demonstrated in the fetuses and dams immediately following abortion. The remaining 4 Brucella-inoculated animals and their fetuses were culture negative for the organism at 14 weeks postinoculation. Results of this study indicate the marine mammal Brucella is capable of producing seroconversion and abortion in cattle but is less pathogenic in that species than B. abortus.
Evaluation of Diagnostic Tests for the Detection of Classical Swine Fever in the Field without a Gold Standard
Authors: Bouma A. , Stegeman J. A. , Engel B. , de Kluijver E. P. , Elbers A. R. W. , de Jong M. C. M.
Published: May 2011

Knowledge of the sensitivity of diagnostic tests for infectious diseases under field conditions can be used to design a surveillance program that increases the effectiveness of the control policy. In this study, the sensitivity of tests for the detection of classical swine fever (CSF) virus (CSFV) under field conditions was estimated without knowledge of the true disease status of the animals tested. During the CSF epidemic of 1997–1998 in The Netherlands, tonsil samples from pigs of CSF suspect farms were collected for laboratory diagnosis of CSF. These specimens were tested in a fluorescence antibody test (FAT1) for the presence of CSFV antigen. When at least 1 specimen in a particular sample series from a farm was positive, this farm was declared CSFV infected. Specimens of that series, either FAT1 negative (98) or FAT1 positive (127), were subsequently tested again (FAT2). After that, a suspension was made of the remaining tissue, and this suspension was evaluated with a virus isolation test. In total, 225 tonsil specimens were examined. A statistical model was formulated, and the sensitivity of the 3 tests and the prevalence of positive specimens in the sample were estimated by the method of maximum likelihood. The sensitivity of the FAT1, the test that was used for confirmation of CSFV infection in a pig herd, was approximately 78% (95% confidence interval [CI] = 62–92%). The effectiveness of the selection process of animals on the farm by the veterinarian was estimated to be 77% (64–87%). The sensitivity of the combination of FAT1 and FAT2 (60%) indicates that at least 5 animals should be selected on a CSF-suspect farm to gain a detection probability for CSFV of 99%.
The Use of a Neutralizing Monoclonal Antibody to Detect Infections of Equine Herpesvirus Type 2 (EHV-2)
Authors: Nordengrahn A. , Klingeborn B. , Lindholm A. , Merza M.
Published: May 2011

A blocking enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies to equine herpesvirus 2 in serum samples of horses. By measuring the binding to a single epitope, this blocking ELISA gives a good picture of the antibody status in the animal. The test is based on a monoclonal antibody with neutralizing activity and had a sensitivity of 94% and a specificity of 100%. Antibodies due to newly acquired infection in foals were successfully detected with this blocking ELISA.
Use of Detergent Extracts of Brucella Abortus RB51 to Detect Serologic Responses in RB51-Vaccinated Cattle
Authors: Hailing Shirley M. , Koster Nancy A.
Published: May 2011

Serologic responses to the newly introduced rough Brucella abortus vaccine strain RB51 have been determined in a dot-blot format using γ-irradiated RB51 cells as the antigen. Because γ-irradiated cells are not easily prepared and the signal from cells was not always reliable, an alternative antigen was sought. Detergent extracts of B. abortus RB51 were prepared using zwittergent 3–14, Triton X-100, and sodium dodecyl sulfate (SDS) and examined in a dot-blot format. Zwittergent 3–14 extracts and γ-irradiated RB51 cells gave the same titers. Unlike γ-irradiated RB51 cells, zwittergent 3–14 extracts produced signals consistently, and the signals were easily interpreted. Triton X-100 extracts interfered with signal development, and SDS extracts resulted in a high background signal. Western blot analyses revealed several outer membrane proteins in the zwittergent 3–14 extract. The major antigens in the extract had apparent molecular weights of <20,000.
Seroprevalence Study of Bovine Neosporosis in Mexico
Authors: Elizabeth Morales S. , Francisco J. Trigo T. , Froylan Ibarra V. , Puente Eduardo C. , Santacruz Mario
Published: May 2011

An indirect enzyme-linked immunosorbent assay was used to obtain epidemiologic information on bovine neosporosis in dairy herds of the Mexican central plateau. Sera were collected from 1,003 cows from 50 dairy herds. Forty-three herds (group A) had been experiencing a high abortion rate. The abortion rates for the remaining 7 herds (group B) were within normal limits for Mexico. Five-hundred sixty-one (56%) of the 1,003 sera were positive. The seroprevalence of Neospora caninum antibodies was 72% (95% Cl = 6875%) in group A and 36% (95% Cl = 31–40%) in group B. These results clearly show that infection with N. caninum is widespread in Mexican dairy herds, as indicated by seropositive cows in group A and group B herds at the time of the sample collection.
Granulomatous Encephalitis in a Neurologically Impaired Goat Kid Associated with Degeneration of Neospora Caninum Tissue Cysts
Authors: Corbellini Luís Gustavo , Colodel Edson Moleta , Driemeier David
Published: May 2011

Congenital Neospora caninum infection was diagnosed in a Saanen goat from a farm in southern Brazil. The kid was unable to nurse and had difficulty rising, ataxia, and opistothotonos. The neurologic signs became more severe 3 days after birth, when it was euthanized. No gross lesions were observed at necropsy. Multifocal infiltrates primarily of mononuclear cells, nodular microgliosis, and perivascular cuffs of lymphocytes, plasma cells, and few neutrophils were observed in the brain, mostly in the cortex and adjacent to ventricles. Rare multinucleate giant cells were observed adjacent to inflammatory foci. Several tissue cysts with a thick wall that reacted strongly with polyclonal antiserum to N. caninum were in the cerebral cortex and medulla oblongata. Lesions were also present in heart, lungs, and liver, but N. caninum tachyzoites were not found. Distinguishing features in this goat kid included neurologic impairment resulting from congenital infection with N. caninum and the presence of granulomatous inflammation with rare giant cells associated with degeneration of tissue cysts.
Haemophilus Somnus Bronchopneumonia in American Bison (Bison Bison)
Authors: Dyer N. W.
Published: May 2011

Immunoperoxidase assays were performed on 21 archived formalin-fixed, paraffin-embedded tissues from from American bison ( Bison bison) with bronchopneumonia. Seven of the 21 bison had positive staining for Haemophilus somnus in alveolar exudate, visceral pleura, lung parenchyma, and chronic necrotic lesions, and H. somnus was isolated from tissues from 1 of these 7 animals. Results suggest that H. somnus is a respiratory pathogen in bison.
Paramyxovirus Infection in Pigs with Interstitial Pneumonia and Encephalitis in the United States
Authors: Janke Bruce H. , Paul Prem S. , Landgraf John G. , Halbur Patrick G. , Huinker Clark D.
Published: May 2011

In the last few years, newly recognized paramyxoviruses have been associated with severe disease in several animal species, including swine, as well as in human beings. Recently, a paramyxovirus was isolated from a swine herd in the northcentral United States that experienced an epizootic of respiratory and central nervous system disease. Affected pigs had interstitial pneumonia with necrotizing bronchiolitis and encephalitis characterized by lymphocytic perivasculitis and diffuse gliosis. Germ-free pigs inoculated with this isolate developed mild clinical illness and similar but less severe histopathologic lesions in lungs and brain.
Equine Monocytic Ehrlichiosis (Potomac Horse Fever) in Horses in Uruguay and Southern Brazil
Authors: Dutra Fernando , Schuch Luíz Filipe D. , Delucchi Eduardo , Curcio Bruna R. , Coimbra Helen , Raffi Margarida B. , Dellagostin Odir , Riet-Correa Franklin
Published: May 2011

A disease named locally as churrío or churrido equino (i.e., equine scours) has occurred for at least 100 years in Uruguay and southern Brazil in farms along both shores of the Merín lake. This report describes cases of churrido equino and provides serologic, pathologic, and DNA-based evidence indicating that the disease is in fact equine monocytic ehrlichiosis (Potomac horse fever). Results of an epidemiological investigation conducted on an endemic farm are also presented. Clinical signs in 12 horses were fever, depression, diarrhea, dehydration, and sometimes colic and distal hind limb edema. Postmortem findings of 3 horses were of acute enterocolitis. Inclusion bodies containing ehrlichial organisms were found in the cytoplasm of macrophages of the large colon of 1 horse. Eleven of the 12 horses were serologically positive to Ehrlichia risticii (indirect fluorescent antibody assay) and, of 3 paired samples, 2 showed seroconversion. Ehrlichia risticii DNA was identified by a nested polymerase chain reaction in peripheral blood of an affected horse. A healthy horse inoculated with peripheral blood from an affected horse developed the disease and antibodies to E. risticii. The disease had a peak incidence in March (summer) and was statistically associated with a marshy ecosystem near the Merín lake, where large numbers of Pomacea spp. (Ampullariidae) snails were found. Incidence density was almost 8 times higher in nonnative horses than in native horses. It was concluded that the previous diarrheic disease of horses known in Uruguay and southern Brazil as churrido equino is equine monocytic ehrlichiosis.
Molecular Identification of Candida Parapsilosis from Crop Mucosa in a Cockatiel
Authors: Kano Rui , Sakamoto Yumi , Hanahachi Ako , Kamata Hiroshi , Fukuda Youichi , Fujiwara Kosaku , Hasegawa Atsuhiko
Published: May 2011

A 2-month-old cockatiel was evaluated for diarrhea, dyspnea, and death. Histologic examination of lesions in the crop mucosa revealed hyperkeratosis and the presence of blastoconidia and hyphae. Positive immunohistochemical staining of the organisms was achieved with an antibody directed against Candida spp. Polymerase chain reaction amplification of DNA from crop lesion material with internal transcribed spacer 2 (ITS2) primers yielded fragments of approximately 300 bp, which demonstrated 95% DNA homology with the corresponding sequence from a strain of Candida parapsilosis deposited in the GenBank data base. The Candida species in the lesion of the crop mucosa was therefore identified by DNA sequence analysis as C. parapsilosis.
Bovine Ephemeral Fever in Taiwan
Authors: Wang Fun-In , Hsu A. M. , Huang K. J.
Published: May 2011

Bovine ephemeral fever (BEF) is a vector-borne disease of cattle, spanning tropical and subtropical zones of Asia, Australia, and Africa, caused by Ephemerovirus of the Rhabdoviridae. Taiwan has had 3 BEF epizootics, occurring in 1989, 1996, and 1999, since the vaccination regimen was initiated in 1984, given once a year in the spring with a single-dose formaldehyde-inactivated vaccine using the 1983 isolate as the seed virus. This study evaluated the 1999 population immunity against BEF virus in Taiwanese dairy cows with a neutralization test and whether the recent BEF virus isolates have mutated significantly from the vaccine virus. In March 1999, before vaccination, 94% of the animals studied were already seropositive, suggestive of an endemic or persistent infection from the previous year. By June 1999, when 51% of herds had been vaccinated, the antibody level rose, and by September 1999, the serum-neutralizing antibody (SNA) level fell to a minimum, preceding the outbreak of BEF in October 1999, during which the antibody levels of vaccinated cows continued to decline while those of unvaccinated cows rose sharply. The results suggest that, in 1999, vaccine-induced immunity was partially protective against BEF. Because the current single-dose vaccination regimen resulted in minimal population immunity by September, a booster vaccination given in late summer may be advisable for future disease control. Analysis of the glycoprotein gene of Taiwanese isolates between 1983 and 1999 showed a 97.4–99.6% homology, with an alteration of 4 amino acids in antigenic sites G1, G3b, and G3c. Phylogenetic analysis of Taiwanese isolates revealed at least 2 distinct clusters: the 1983–1989 isolates and the 1996–1999 isolates. Both were distinct from 2 Japanese strains and the Australian BB7721 strain. Thus, at least 2 distinct BEF viruses, which had diverged before 1983, existed in Taiwanese dairy cows.
Prevalence of Virulent Rhodococcus Equi in Soil from Five R. Equi-Endemic Horse-Breeding Farms and Restriction Fragment Length Polymorphisms of Virulence Plasmids in Isolates from Soil and Infected Foals in Texas
Authors: Takai Shinji , Chaffin M. Keith , Cohen Noah D. , Hara Michiyo , Nakamura Mutsu , Kakuda Tsutomu , Sasaki Yukako , Tsubaki Shiro , Martens Ronald J.
Published: May 2011

Rhodococcus equi isolates (462) obtained from 64 soil samples collected on 5 R. equi-endemic horse-breeding farms and isolates from 100 infected foals in Texas were examined to determine the prevalence and genotypic diversity of virulence-associated plasmids. Isolates were tested for the presence of 15–17-kDa virulence-associated protein antigens (VapA) by immunoblotting and virulence-associated plasmids by PCR. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases for estimation of size and comparison of polymorphisims. Rhodococcus equi were isolated from soil of all 5 farms; however, virulent R. equi were only isolated from 3 of the 5 farms and represented 18.8% (87 of 462) of total isolates. Of the 87 virulent soil isolates, 56 (64.5%) contained an 85-kb type I plasmid, 23 (26.4%) an 87-kb type I plasmid, 7 (8%) a newly defined 85-kb type III plasmid (Tx 43), and 1 (1.1%) a newly defined 85-kb type IV plasmid (Tx 47). Of the 100 isolates from infected foals, 96 were virulent. Of the 96 virulent isolates, 51 (53.1%) contained an 85-kb type I plasmid, 39 (40.6%) an 87-kb type I plasmid, 4 (4.2%) an 85-kb type III plasmid (Tx 43), and 2 (2.1%) an 85-kb type IV plasmid (Tx 47). There are at least 4 different R. equi virulence-associated plasmids in Texas, 2 of which have not previously been described. Based upon virulence plasmid typing, there is geographic diversity among isolates of R. equi from clinical and environmental samples on horse-breeding farms in Texas. There is not a strong correlation between the presence of virulent R. equi in farm soils and the R. equi disease status of those farms.
Endometritis in Postparturient Cattle Associated with Bovine Herpesvirus-4 Infection: 15 Cases
Authors: Frazier Ken , Pence Mel , Mauel Michael J. , Liggett Alan , Hines Murray E. , Sangster Lowell , Lehmkuhl Howard D. , Miller Debra , Styer Eloise , West Joe , Baldwin Charles A.
Published: May 2011

Suppurative, ulcerative endometritis associated with bovine herpesvirus-4 (BHV-4) infection was identified in 15 postparturient dairy cows from 5 separate dairies. Characteristic eosinophilic to amphophilic intranuclear viral inclusion bodies were identified within degenerate endometrial lining epithelium and endothelial cells. Bovine herpesvirus-4 was confirmed as the etiology by a combination of fluorescent antibody assays, viral isolation, heminested PCR, ultrastructural examination of the uterus and inoculated tissue culture cells, and negative-stain electron microscopy of tissue culture supernatant. Viral particles measuring 70–95 nm were demonstrated in uterine epithelial and endothelial cells by electron microscopy. Bacteria including Arcanobacterium pyogenes, Escherichia coli, and an α- Streptococcus isolate were isolated from all uteri. Bovine herpesvirus-4-associated endometritis has been previously reported in sporadic cases in Europe but has not been previously reported in the United States. Endometritis associated with BHV-4 appears to be an emerging syndrome in Georgia dairy herds.
Application of a PCR Assay to Enhance the Detection and Identification of Tritrichomonas Foetus in Cultured Preputial Samples
Authors: Parker Sarah , Lun Zhao-Rong , Gajadhar Alvin
Published: May 2011

The traditional diagnostic test for Tritrichomonas foetus involves collection of preputial or vaginal samples followed by culture in a growth media and microscopic examination. Recently, polymerase chain reaction (PCR) techniques have been described for use as a diagnostic assay. The objective of this study was to evaluate a previously described PCR assay for detecting T. foetus in cultured preputial material. The detection limits of the assay for T. foetus organisms in a growth medium, in samples prepared from washing microscope slides, and in preputial material cultured in a growth medium were determined. Preputial samples were collected from 13 bulls uninfected with T. foetus. The PCR assay was able to detect 5 T. foetus organisms in the growth medium and the cultured preputial material. Amplification products were obtained from samples prepared from washes of microscope slides containing as few as 3 visualized organisms. The PCR assay was able to detect organisms in culture at a lower concentration than was possible by direct microscopic examination. This low detection limit may allow the PCR assay to be used to enhance the sensitivity of the current diagnostic test. In addition, the assay could be used to confirm the identification of T. foetus organisms observed by direct microscopic examination when other confirmation techniques, such as staining and phase microscopy, are not practical.
Differential Detection of Transmissible Gastroenteritis Virus and Porcine Epidemic Diarrhea Virus by Duplex RT-PCR
Authors: Kim So Y. , Song Dae S. , Park Bong K.
Published: May 2011

Transmissible gastroenteritis (TGE) and porcine epidemic diarrhea (PED) are highly contagious enteric diseases of piglets. The clinical signs of these diseases are very similar and include watery, yellowish diarrhea. Thus, the effective differential detection of TGE virus and PED virus is required. In the present study, a duplex reverse transcription-polymerase chain reaction (RT-PCR) was established for the differential detection of TGE and PED viruses. The primers were designed for the S gene of each virus. RNA was extracted from the intestines and stool samples that were collected from the swine with diarrhea. The RT-PCR test could detect both TGE and PED viruses with 2 TCID50/200 μl. Among 90 clinical samples, 7 TGE viruses and 2 PED viruses were detected by the duplex RT-PCR. This duplex RT-PCR may be a useful diagnostic method for the rapid, specific, and sensitive differential detection of TGE and PED viruses using clinical samples.
Hydatidosis in a Chapman's Zebra (Equus Burchelli Antiquorum)
Authors: Chiou Ming T. , Wang Fun-In , Chang Pen H. , Liu Chen-Hsuan , Jeng Chian R. , Cheng Chiung H. , Jou Jiann , Pang Victor F.
Published: May 2011

A 14-year-old male Chapman's zebra had been treated for a cervical granulomatous cellulitis for 3 months prior to sudden death associated with myocardial fibrosis. Incidental findings at necropsy included multiple 3–8-cm unilocular cysts in the liver and lungs. Cysts contained either a clear fluid with or without hydatid sand or a gelatinous substance with foci of mineralization. Light and scanning electron microscopic examination of the hydatid sand revealed free protoscolices as well as intact and ruptured brood capsules with protoscolices attached to the germinal membrane. The protoscolices had 2 rows of 36–38 rostellar hooks with a length of 25–30 μm. The cyst wall consisted of inner germinal, intermediate laminated, and outer adventitial layers. Hydatidosis caused by Echinococcus granulosus was diagnosed based on the unilocular cysts, multiple protoscolices formed in a brood capsule, typical trilayered cyst wall, and herbivorous intermediate host. This is the first reported case of animal hydatidosis in Taiwan. The infection is thought to have been established 12 years ago in South Africa prior to importation.
Validation of Monoclonal Antibody F99/97.6.1 for Immunohistochemical Staining of Brain and Tonsil in Mule Deer (Odocoileus Hemionus) with Chronic Wasting Disease
Authors: Spraker T. R. , O'Rourke K. I. , Balachandran A. , Zink R. R. , Cummings B. A. , Miller M. W. , Powers B. E.
Published: May 2011

A new monoclonal antibody (MAb), F99/97.6.1, that has been used to demonstrate scrapie-associated prion protein PrPsc in brain and lymphoid tissues of domestic sheep with scrapie was used in an immunohistochemistry assay for diagnosis of chronic wasting disease (CWD) in mule deer ( Odocoileus hemionus). The MAb F99/97.6.1 immunohistochemistry assay was evaluated in brain and tonsil tissue from 100 mule deer that had spongiform encephalopathy compatible with CWD and from 1,050 mule deer outside the CWD-endemic area. This MAb demonstrated abnormal protease-resistant prion protein (PrPres) in brains of all of the 100 mule deer and in 99 of the 100 tonsil samples. No immunostaining was seen in samples collected from deer outside the endemic area. MAb F99/97.6.1 demonstrated excellent properties for detection of PrPres in fresh, frozen, or mildly to moderately autolytic samples of brain and tonsil. This immunohistochemistry assay is a sensitive, specific, readily standardized diagnostic test for CWD in deer.
An Improved Immunohistochemical Diagnostic Technique for Canine Leptospirosis using Antileptospiral Antibodies on Renal Tissue
Authors: Wild Chad J. , Greenlee Justin J. , Bolin Carole A. , Barnett Jeanne K. , Haake A. David , Cheville Norman F.
Published: May 2011

The purpose of this study was to compare the immunoreactivity in canine renal tissues stained with antisera specific for 3 leptospiral antigens and those processed with traditional staining methods. In addition, immunoglobulin staining was done on tissues with immunoreactivity to leptospiral antigens. Formalin-fixed renal sections from 12 dogs with chronic interstitial nephritis suspected or proven to have leptospirosis (6 dogs with silver-stained leptospires and 6 dogs in which silver-stained leptospires were not detected) were used. Antibodies consisted of a monoclonal antibody to Leptospira kirschneri serovar grippotyphosa lipopolysaccharide (LPS) and 2 polyclonal antibodies to outer membrane proteins, including OmpL1, a leptospiral porin, and LipL41, an outer membrane lipoprotein. The murine monoclonal antisera against LPS (F71C2-1) had the most abundant and consistent immunoreactivity. Immunoreactive areas were present in 6 of 6 sections positive by silver staining and included extracellular granular debris in intertubular areas, debris in macrophages, organisms in tubular lumina, and cytoplasmic granules in tubular epithelia. Antisera with specificity for the outer membrane proteins OmpL1 and LipL41 detected only intact organisms in tubular lumina. Immunoreactivity to OmpL1 (polyclonal 338) occurred in 4 of 5 sections positive by silver staining, but immunoreactivity to LipL41 (polyclonal 813) occurred in only 1 of 6 silver-positive sections. Each of the kidney sections in which leptospiral antigens were detected by immunohistochemistry also was positive by silver staining. Sections negative by silver staining were also negative by immunostaining. Although immunohistochemistry did not enhance sensitivity, amplification of signal by secondary antibody and hematoxylin counterstaining improved the ease of diagnosis and allowed better evaluation of tissue morphology than did silver staining methods. IgG was the most abundant immunoglobulin. IgG immunoreactivity occurred predominantly in plasma cells within interstitial infiltrates. Interstitial infiltrates contained abundant immunoreactivity to LPS, but immunoreactivity to OmpL1 and LipL41 was not noted.
Polymerase Chain Reaction and DNA Sequencing for Detection of Ovine Herpesvirus 2 in American Bison (Bison Bison)
Authors: Sausker Elizabeth A. , Dyer Neil W.
Published: May 2011

Serum samples were collected at slaughter from 226 24–30-month-old ranch-raised, clinically normal American bison ( Bison bison) bulls from North Dakota, Minnesota, Kansas, and Manitoba to assess the presence of antibodies to ovine herpesvirus 2 (OHV-2). Antibodies to OHV-2 were detected by competitive inhibition enzyme-linked immunosorbent assay in 10 of 226 (4.40%) samples. Polymerase chain reaction (PCR) analysis of sera positive for OHV-2 DNA demonstrated a 238 kilobase fragment. The nucleotide sequence of the PCR-positive samples in comparison to the reported OHV-2 nucleotide sequence resulted in a homology range of 82.8–95.4%.
Altered Immunohistochemical Staining for Desmoglein in Skin Biopsies in Canine Pemphigus Foliaceus
Authors: Steeves Elizabeth B. , Chelack Brian J. , Clark Edward G. , Haines Deborah M.
Published: May 2011

In this study, 50 cases of canine pemphigus foliaceus and 49 cases of canine superficial pyoderma were examined by immunohistochemical staining for patterns of desmoglein expression. In 31/50 (62%) of pemphigus foliaceus cases, there was an altered staining pattern for desmoglein consisting of distinct clumped deposits at the periphery of keratinocytes and/or dark cytoplasmic staining of acantholytic cells (consistent with internalization of desmoglein). In contrast, desmoglein staining in biopsies from cases of superficial pyoderma was diffusely pale without evidence for clumping or distinct internalization. This study demonstrates that epidermal desmoglein expression is altered in some cases of pemphigus foliaceus in dogs and suggests that immunohistochemical staining for this protein may be useful in diagnosis.
Evaluation of a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay for Subtyping Hemagglutinin Genes 1 and 3 of Swine Influenza Type A Virus in Clinical Samples
Authors: Choi Young Ki , Goyal Sagar M. , Joo Han Soo
Published: May 2011

A multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay was developed to detect and identify subtypes of hemagglutinin (H) 1 and H3 swine influenza virus (SIV). Two oligonucleotide primer sets were prepared using published sequence data for H1N1 and H3N2. The PCR products with unique size characteristics of each subtype were sequenced, and the sequences were confirmed to be subtype specific for H gene 1 or 3. These primer sets did not amplify when RT-PCR assay was performed for genomic DNA or RNA from other common swine pathogens. The RT-PCR assay was able to detect viral RNA up to 1 tissue culture infective dose of reference SIV H1N1 or H3N2. The multiplex RT-PCR was applied to 30 SIV isolates subtyped by hemagglutination inhibition (HI) test. Forty-three positive and 20 negative swine field samples for SIV by virus isolation were also tested. Of these 73 SIV-positive samples tested, H1 and H3 were identified in 38 and 28 samples by the multiplex RT-PCR, respectively. The remaining 7 samples were positive for both H1 and H3 genes. No positive reaction was found for all 20 SIV-negative field samples. Subsequently, 235 random field samples from pigs with respiratory problems were tested by the multiplex RT-PCR, and 26 and 13 samples were found to be positive for H1 and H3, respectively. Results of these multiplex RT-PCR were comparable with those of the HI test. These results suggest that multiplex RT-PCR can be a useful test for detection and subtyping of SIV in clinical samples.
Seroprevalence of OHV-2, BVDV, BHV-1, and BRSV in Ranch-Raised Bison (Bison Bison)
Authors: Sausker Elizabeth A. , Dyer Neil W.
Published: May 2011

Serum samples were collected at slaughter from 226 24–30-month-old American bison ( Bison bison) bulls from Kansas, Minnesota, North Dakota, and Manitoba and assayed for antibodies to ovine herpesvirus type-2 (OHV-2), bovine viral diarrhea virus (BVDV), bovine herpesvirus type-1 (BHV-1), and bovine respiratory syncytial virus (BRSV). Antibodies were detected by serum neutralization for BVDV, BHV-1, and BRSV, while antibodies to OHV-2 were detected by competitive inhibition-ELISA (CI-ELISA). Detectable antibodies were found against all viruses: 10 of 226 (4.40%) against OHV-2, 125 of 226 (55.3%) against BVDV, 99 of 226 (43.8%) against BHV-1, and 208 of 226 (92.0%) against BRSV. Titers from 93.6% of the BVDV-positive animals, 79.8% of the BHV-1-positive animals, and 98.1% of the BRSV-positive animals were ≥ 1.25. These data indicate that a low percentage of clinically normal bison are seropositive for OHV-2 while a high percentage of bison sampled are seropositive for BVDV, BHV-1, and BRSV.
Pyogranulomatous Meningoencephalitis Associated with Dematiaceous Fungal (Cladophialophora Bantiana) Infection in a Domestic Cat
Authors: Bouljihad Mostafa , Lindeman Cindy J. , Hayden David W.
Published: May 2011

A 6-month-old, castrated male domestic cat with progressive neurological signs of 2–3 weeks duration was necropsied. Macroscopic findings were restricted to the brain and included irregularly shaped, well-delineated but unencapsulated areas of intense black pigmentation involving the rostral portion of both cerebral hemispheres. Microscopically, numerous brown, oblong, segmented branching hyphae and conidial-like structures and extensive pyogranulomatous inflammation were identified throughout the cerebral lesion and in adjacent blood vessels. Hyphae and oval conidia were best demonstrated with either Gomori methenamine silver or periodic acid–Schiff stain. Fungal infection in the brain of this cat was unrelated to any concurrent immunodeficiency syndrome or immunosuppressive treatment. This report deals with a case of cerebral phaeohyphomycosis from which a different species of dematiaceous fungus, Cladophialophora bantiana, was isolated and identified.
Failure of Porcine Reproductive and Respiratory Syndrome Virus to Replicate in Porcine Endothelial Cell Cultures
Authors: Howerth E. W. , Murphy M. D. , Roberts A. W.
Published: May 2011

Clinical, gross, and microscopic pathologic and immunohistochemical findings in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) suggest that PRRSV may replicate in endothelial cells. Endothelial cell cultures from porcine aorta and pulmonary artery were tested for susceptibility to various strains of PRRSV. Cultures were identified as endothelium by light microscopy and immunohistochemical staining for P-selectin and von Willebrand factor. Five strains of PRRSV, i.e., the National Veterinary Services Laboratories, Ames, IA PRRSV strain 130-PDV and 4 field strains isolated from pneumonic lungs, failed to replicate in these porcine large-vessel endothelial cell cultures.
The Age of Competence: An Update on the International Laboratory Accreditation Scene for Veterinary Testing Laboratories
Authors: Wiegers Ann L.
Published: May 2011

Many changes have recently taken place in the world of laboratory accreditation. These changes include the increased use of voluntary standards in lieu of regulations, a move towards harmonization (equivalent results using voluntary standards) over standardization (using the same procedures or regulations), and an increased focus on competence, which includes the competence of laboratories to conduct testing, the competence of accreditation bodies to operate accreditation programs, the competence of bodies such as the National Cooperation for Laboratory Accreditation (NACLA) to recognize accreditation bodies as meeting the requirements of relevant standards, and the competence of organizations providing services to the accreditation process, such as the operation of proficiency testing programs. To describe these changes, a brief and general description of the International Laboratory Accreditation Cooperation accreditation scheme is provided, including an update on relevant decisions and activities in the United States and a description of the organization and activities of the newly formed NACLA. Following this discussion, with emphasis on veterinary testing, is an overview of several national and international organizations, including accreditation bodies, that promote harmonization, standardization, and analytical excellence. Also outlined are relevant activities of these organizations, an overview of some of the standards and guidelines they produce, and a description of how such organizations interact with each other and with laboratories seeking recognition for competence. Next is a brief discussion of recent developments and trends in laboratory accreditation, the impact of these developments, and the costs and benefits of accreditation to laboratories. Suggestions to veterinary laboratories for formulating strategy for keeping current with developments in accreditation and for determining quality goals are included.
Field Validation of a Commercial Blocking ELISA to Differentiate Antibody to Transmissible Gastroenteritis Virus (TGEV) and Porcine Respiratory Coronavirus and to Identify TGEV-Infected Swine Herds
Authors: Carman Susy , Josephson Gaylan , McEwen Beverly , Maxie Grant , Antochi Mioara , Eernisse Ken , Nayar Gopi , Halbur Pat , Erickson Gene , Nilsson Ernst
Published: May 2011

A commercially available blocking ELISA was analyzed for its ability to identify antibodies to porcine coronaviruses (transmissible gastroenteritis virus [TGEV] or porcine respiratory coronavirus [PRCV]), to differentiate antibodies to TGEV and PRCV, and to identify TGEV-infected herds. Nine sera from uninfected pigs, 34 sera from 16 pigs experimentally infected with TGEV, and sera from 10 pigs experimentally infected with PRCV were evaluated using both the TGEV/PRCV blocking ELISA and a virus neutralization (VN) assay. The ELISA was not consistently effective in identifying pigs experimentally infected with TGEV until 21 days postinfection. Sera from 100 commercial swine herds (1,783 sera; median 15 per herd) were similarly evaluated using both tests. Thirty of these commercial herds had a clinical history of TGEV infection and a positive TGEV fluorescent antibody test recorded at necropsy within the last 35 months, while 70 herds had no history of clinical TGEV infection. The blocking ELISA and the VN showed good agreement (kappa 0.84) for the detection of porcine coronavirus antibody (TGEV or PRCV). The sensitivity (0.933) of the ELISA to identify TGEV-infected herds was good when considered on a herd basis. The ELISA was also highly specific (0.943) for the detection of TGEV-infected herds when the test results were evaluated on a herd basis. When sera from specific age groups were compared, the ELISA identified a greater proportion (0.83) of pigs in herds with TGEV antibody when suckling piglets were used. In repeatability experiments, the ELISA gave consistent results when the same sera were evaluated on different days (kappa 0.889) and when sera were evaluated before and after heating (kappa 0.888). The blocking ELISA was determined to be useful for herd monitoring programs and could be used alone without parallel use of the VN assay for the assessment of large swine populations for the detection of TGEV-infected herds.
Evaluation of a Porcine Circovirus Type 2–Specific Antigen-Capture Enzyme-Linked Immunosorbent Assay for the Diagnosis of Postweaning Multisystemic Wasting Syndrome in Pigs: Comparison with Virus Isolation, Immunohistochemistry, and the Polymerase Chain Reaction
Authors: McNeilly F. , McNair I. , O'Connor M. , Brockbank S. , Gilpin D. , Lasagna C. , Boriosi G. , Meehan B. , Ellis J. , Krakowka S. , Allan G. M.
Published: May 2011

Quantitative virus isolation, immunohistochemistry, polymerase chain reaction (PCR) assay, and a porcine circovirus 2 (PCV2)-specific antigen-capture enzyme-linked immunosorbent assay (ELISA) were used for differentiation between clinical and subclinical PCV2 infections of swine. Tissue samples from pigs experimentally infected with PCV2 and field cases of postweaning multisystemic wasting syndrome and PCV2-associated reproductive disorders were used in this evaluation. In initial studies on 6 PCV2 pools using 3 previously published PCR protocols for PCV2 detection, quantitative virus isolation, and antigen-capture ELISA, substantial differences in sensitivity were identified among these procedures. Examination of tissue samples from diseased and clinically normal pigs indicated that immunohistochemistry, quantitative virus isolation, and antigen-capture ELISA could be used to differentiate between clinical and subclinical PCV2 infections, but the PCR assay could not. Because subclinical infections of pigs with PCV2 are common, the use of nonquantitative PCR as a diagnostic tool for PCV2-related diseases should be discouraged and the PCV2-specific antigen-capture ELISA evaluated further.
Primary Epitheliotropic Intestinal T-cell Lymphoma in a Horse
Authors: Pinkerton Marie E. , Bailey Keith L. , Thomas Kathy K. , Goetz Thomas E. , Valli Victor E.
Published: May 2011

A 17-year-old Quarterhorse gelding with a clinical diagnosis of protein-losing enteropathy was submitted for necropsy following a 4–5-month duration of weight loss, decreased appetite, and hypoproteinemia. Gross findings included multiple 1–2-cm diameter ulcers on the luminal surfaces of the duodenum and ileum. Histologic examination revealed individual large, round cells infiltrating much of the mucosal epithelium of the duodenum, jejunum, ileum, and colon in addition to multifocal areas of ulceration. Similar round cells infiltrated Brunner's glands and expanded the submucosa beneath the foci of ulceration. Immunohistochemical staining indicated the round cell population was of T-lymphocyte origin. Several features of this equine neoplasm bear similarities to enteropathy-associated T-cell lymphoma in humans.
Antimicrobial Susceptibility of Actinobacillus Pleuropneumoniae, Escherichia Coli, and Salmonella Choleraesuis Recovered from Taiwanese Swine
Authors: Chang Chao-Fu , Chang Lin-Chu , Chang Yung-Fu , Chen Michael , Chiang Tai-Sheng
Published: May 2011

Minimum inhibition concentrations (MICs) were determined for ampicillin, ceftiofur, cephalothin, chloramphenicol, enrofloxacin, gentamicin, lincomycin, lincospectin (lincomycin/spectinomycin), neomycin, premafloxacin, spectinomycin, sulfamethoxazole/trimethoprim, and tetracycline against a total of 180 isolates of Actinobacillus pleuropneumoniae, Escherichia coli, and Salmonella choleraesuis (60 each) clinically isolated from pigs on farms in Taiwan from 1994 to 1996. No more than 3 isolates per farm were used. Ceftiofur had the highest activity in vitro against isolates of A. pleuropneumoniae, E. coli, and S. choleraesuis, with MIC90 values of 0.03, 2, and 1 μg/ml, respectively. Premafloxacin was highly active against isolates of A. pleuropneumoniae, E. coli, and S. choleraesuis, with MIC90 values of 2, 8, and 0.5 μg/ml, respectively, which were lower than those with enrofloxacin (MIC90 8, 32, and 2 μg/ml, respectively). Neomycin was moderately active against A. pleuropneumoniae and E. coli, with MIC90 values of 8 and 64 μg/ml, respectively, but was inactive with S. choleraesuis. Gentamicin showed high activity against A. pleuropneumoniae (MIC90 of 2 μg/ml) but was only moderately active with E. coli and S. choleraesuis (MIC90 of 64 and 32 μg/ml). Cephalothin was highly active against isolates of A. pleuropneumoniae (MIC90 of 1 μg/ml) but was inactive with E. coli (MIC90 of 128 μg/ml). Lincomycin had moderate activity (MIC90 of 32 μg/ml) against A. pleuropneumoniae. Chloramphenicol, lincomycin, and tetracycline were inactive with E. coli and S. choleraesuis (MIC90 > 128 μg/ml). In conclusion, ceftiofur and premafloxacin were highly active against isolates of A. pleuropneumoniae, E. coli, and S. choleraesuis; enrofloxacin and gentamicin were highly to moderately active; cephalothin was highly active against A. pleuropneumoniae and moderately active against S. cholearesuis; chloramphenicol, lincomycin, and tetracycline were active only with A. pleuropneumoniae; neomycin was moderately active against A. pleuropneumoniae and E. coli. The other antimicrobials tested were inactive.
An Immunoperoxidase Monolayer Assay for the Detection of Antibodies against Swine Influenza Virus
Authors: Direksin Kochakorn , Joo HanSoo , Goyal Sagar M.
Published: May 2011

An immunoperoxidase monolayer assay (IPMA) has been developed to detect antibodies against swine influenza A virus (SIV) in pig sera. The test was evaluated by using sequential sera from pigs experimentally infected with H1N1 subtype of SIV. Two hundred field serum samples that had been examined by the hemagglutination inhibition (HI) test were also tested. Antibodies specific to SIV were detected as early as 3 days postinoculation (dpi) in the IPMA test as compared with 7 dpi by the HI test. Unlike HI, no serum treatment was required in the IPMA test. Regardless of the virus used in the test, IPMA detected antibodies to both H1N1 and H3N2 subtypes of SIV whereas HI detects antibodies against either H1N1 or H3N2, depending upon the virus used in the test. Results of this study indicate that IPMA is a useful test for screening of pig sera for SIV antibodies.
Neuroaxonal Dystrophy in Raccoons (Procyon Lotor) from Iowa
Authors: Hamir Amir N. , Miller Janice M. , Stack Mick J. , Chaplin Melanie J. , Cutlip Randall C.
Published: May 2011

During a 12-month period (1998–1999), microscopic evidence of neuroaxonal dystrophy (NAD) in medullae oblongata of raccoons ( Procyon lotor) was observed in 17/39 (47% prevalence in adults) from Iowa, USA. Three of the animals were kits (<3 months), 26 were between 1 and 2 years, and 10 were over 7 years. Lesions were not seen in the medullae of the 3 kits. In young adults, the lesions were mild and were seen in 7 animals. More severe lesions were present in the 10 older raccoons. Grossly, the brains were unremarkable. Microscopically, NAD was confined to the dorsal caudal medulla, where certain nuclei (predominantly gracilis and cuneate) were bilaterally affected. Severely affected animals had vacuolar degeneration of neurons or neuronal loss and extensive areas of spongiosis. Tests for the presence of PrPres in the brain were negative. Spongiotic areas often contained axonal spheroids. Degenerate neurons and axons occasionally contained amphophilic periodic acid-Schiff-positive granular material. There was a paucity of inflammatory cells in the affected areas. Since lesions were not present in kits, were either absent or mild in young adults, and were severe in older raccoons, the findings may be related to advancing age. Neuroaxonal dystrophy has not been previously reported in raccoons. Retrospective examination of raccoon brains from the eastern and northwestern areas of the country revealed very low prevalence of NAD. Because of the apparently high prevalence of this condition at this geographic location, factors other than age (genetic, nutritional, and/or environmental) may influence this degenerative process in the brains of raccoons in Iowa.
Malignant Catarrhal Fever in a Bison (Bison Bison) Feedlot, 1993–2000
Authors: O'Toole D. , Li H. , Sourk C. , Montgomery D. L. , Crawford T. B.
Published: May 2011

A fatal enteric syndrome was identified in American bison ( Bison bison) at a large feedlot in the American Midwest in early 1998. An estimated 150 bison died of the syndrome between January 1998 and December 1999. The syndrome was identified as malignant catarrhal fever (MCF), primarily the alimentary form. Clinical onset was acute, and most affected bison died within 1–3 days; none recovered. Consistent lesions were hemorrhagic cystitis, ulcerative enterotyphlocolitis, and arteritis-phlebitis. Vasculitis was milder and more localized than that in cattle with MCF, and in contrast to the situation in cattle, lymphadenomegaly was minimal. Virtually all affected bison examined were positive for ovine herpesvirus 2 (OvHV-2) by polymerase chain reaction (PCR) assay. A retrospective study of archived tissues established that MCF occurred in the yard as early as 1993. A prospective study was undertaken to establish the importance of MCF relative to other fatal diseases at the feedlot. The fate of a group of 300 healthy male bison in a consignment of 1,101 animals was followed for up to 7 months to slaughter. At entry, 23% (71/300) of bison were seropositive for MCF viruses, and 11% (8/71) of these seropositive bison were PCR positive for OvHV-2. Forty seronegative bison were selected at random from the group, and all were PCR negative for OvHV-2. There was no change in seroprevalence in the group during the investigation. The minimum infection rate for MCF virus was 36.3% (93/256). Twenty-two (7.3%) of the 300 bison in the feedlot died. Of these, 15 had MCF, 4 had acute or chronic pneumonia, and 3 were unexamined. Losses in the entire consignment were higher (98/1,101; 8.8% death loss); 76% of deaths were attributable to MCF. The study failed to reveal a relationship between subclinical infection and development of clinical disease.