bluuebunny/crossref_metadata_2025 huggingface dataset:
Otitis Media in Preweaned Holstein Dairy Calves in Michigan Due to Mycoplasma Bovis
Authors: Walz Paul H. , Mullaney Thomas P. , Render James A. , Walker Robert D. , Mosser Theresa , Baker John C.
Published: May 2011

Mycoplasma bovis was isolated from the tympanic bullae of dairy calves with an exudative otitis media. The history, clinical signs, gross and histologic lesions, and bacteriologic findings are described for 5 preweaned Holstein calves with otitis media from a 600-cow dairy in Michigan. Clinical findings consisted of unilateral or bilateral ear droop, epiphora, head tilt, and recumbency in severely affected calves. Postmortem examination revealed unilateral or bilateral fibrinosuppurative to caseous exudate in the tympanic bullae. Histologically, a marked fibrinosuppurative to caseous exudate filled the tympanic air spaces. The partially ulcerated tympanic mucosa was markedly thickened with mononuclear cell infiltration and proliferation of fibrous connective tissue. Bone remodeling and periosteal hyperostosis were present in some osseous septa. Mycoplasma bovis was isolated from the tympanic bullae of all 5 calves and from the lungs of 2 calves and the frontal sinus of 1 calf. Mycoplasma bovis was isolated at > 100,000 colony forming units/ml from the bulk milk tank of the farm of origin. The isolation of M. bovis from the bulk milk tank, indicating subclinical mycoplasmal mastitis coupled with the feeding of waste milk from mastitic cows to calves is suggestive of a possible source of the infection resulting in otitis media in preweaned dairy calves.
Detection of Canine Parvovirus in Naturally Infected Dogs with Enteritis and Myocarditis by in Situ Hybridization
Authors: Nho Whan-Gook , Sur Jung-Hyang , Doster Alan R. , Kim Soon-Bok
Published: May 2011

An improved method for the diagnosis of canine parvovirus using in situ hybridization in standard formalin-fixed, paraffin-embedded tissue sections was developed. A digoxigenin-labeled probe complementary to DNA sequences that code for the entire sequence of the capsid protein VP-1 and the middle part of the sequence of the capsid protein VP-2 was designed. Specific histologic localization of canine parvovirus-infected cells was demonstrated in small intestine, tonsil, lymph node, thymus, spleen, heart, liver, and kidney from dogs diagnosed at necropsy with canine parvovirus infection. The in situ hybridization accurately pinpointed the specific sites of viral infection. The detection of canine parvovirus in liver, kidney, and heart tissues together in the same pups could represent an enhanced virulence of this strain of canine parvovirus and suggests a broadened tissue tropism not seen before in Korean strains of canine parvovirus.
Validation of Nonradioactive Chemiluminescent Immunoassay Methods for the Analysis of Thyroxine and Cortisol in Blood Samples Obtained from Dogs, Cats, and Horses
Authors: Singh A. K. , Jiang Y. , White T. , Spassova D.
Published: May 2011

The performances of a radioimmunoassay method, a chemiluminescent immunoassay method, and a chemiluminescent-enzyme immunoassay method were evaluated for the analysis of cortisol and total thyroxine in blood samples obtained from dogs, cats, horses, and humans (reference samples). The analysis of cortisol in human and animal samples exhibited good precision, linearity, and recovery. The 3 methods gave comparable values for the ACTH-induced increase and the dexamethasone-induced decrease in cortisol concentrations in animal samples. The recoveries of total thyroxine from human samples, analyzed by the 3 methods, were comparable. However, the basal total thyroxine concentrations determined by the chemiluminescent immunoassay method were 30–40% lower than those determined by the radioimmunoassay and the chemiluminescent-enzyme immunoassay methods in animal samples. In both human and animal samples, the plot of thyroxine values obtained by the radioimmunoassay method against those obtained by the chemiluminescent immunoassay method or the chemiluminescent-enzyme immunoassay method was linear. However, although the slope of the radioimmunoassay versus chemiluminescent-enzyme immunoassay curve was close to unity, the slope of the radioimmunoassay versus chemiluminescent immunoassay curve was 0.6. This result suggests that, compared with the radioimmunoassay method, the chemiluminescent immunoassay method underestimated thyroxine values in animal samples but not in human samples. Although all 3 methods yielded comparable changes in thyroxine concentrations in response to thyroid stimulating hormone, they did not yield comparable thyroxine concentrations in response to T3 suppression in dogs and cats.
Comparison of Conventional Culture Methods and Two Commercial Enzyme Immunoassays for Detection of Salmonella in Porcine Fecal Samples and Cecal Contents
Authors: Wegener Henrik Caspar , Baggesen Dorte Lau
Published: May 2011

Two commercial enzyme immunoassays, designated EIA-1 and EIA-2, for the detection of salmonella in feces and cecal contents were compared to conventional culture methods. Out of 362 cecal content samples, 35 were positive by EIA-1 and 30 were positive by EIA-2 and conventional methods. Out of 189 fecal samples, 41 were positive by EIA-1, 30 were positive by EIA-2, and 24 were positive by conventional culture methods. The EIA-1 assay method detected more positive samples overall, although only comparison of the fecal samples was statistically significant. The results indicate that the EIA-1 method is rapid, requiring only 24 hours, and may be more sensitive than the EIA-2 assay and conventional culture techniques. Enzyme immunoassays for the detection of salmonella in fecal samples or cecal contents provide a reasonable alternative to conventional culture techniques.
Identification of Mycoplasma Iowae by Colony Immunoblotting Utilizing Monoclonal Antibodies
Authors: Singh P. , Yavari Christine A. , Newman John A. , Bradbury Janet M.
Published: May 2011

The utility of monoclonal antibody Mab-2C in identification of Mycoplasma iowae (MI) by colony immunoblotting technique was explored. Colony immunoblots of reference MI strains, field isolates, and mycoplasmas recovered from experimentally inoculated turkey embryos were probed with Mab-2C. The monoclonal antibody identified colonies of all the MI isolates tested and did not cross-react with colonies of M. gallisepticum, M. synoviae, or M. meleagridis. In western immunoblots of 22 MI field isolates, Mab-2C showed immunoreactivity with an antigen of approximately 45 kD molecular weight. No phenotypic variation of the epitope recognized by Mab-2C was observed in colony immunoblots of MI colonies. The monoclonal antibody reported here can be used for identification of MI colonies by a simple and rapid colony immunoblot method.
Seminal Vesiculitis and Orchitis Caused by Brucella Abortus Biovar 1 in Young Bison Bulls from South Dakota
Authors: Rhyan Jack C. , Holland Sam D. , Gidlewski Thomas , Saari Dennis A. , Jensen Allen E. , Ewalt Darla R. , Hennager Steve G. , Olsen Steven C. , Cheville Norman F.
Published: May 2011

Specimens of blood, lymph nodes, spleens, and genitalia were collected at slaughter from seven 3- and 4-year-old male bison that had recently become seropositive for brucellosis. The animals were from a captive herd of approximately 3,500 bison located in central South Dakota. Brucella abortus biovar 1 was isolated from 2 or more specimens from each of 6 bison. Severe necrotizing and pyogranulomatous orchitis was present in 1 testicle from 1 bull, and 4 animals had mild to marked seminal vesiculitis. Immunohistochemical staining labeled organisms in seminal vesicles and the testicle with orchitis. Ultrastructurally, intact bacilli were present in cytoplasmic vacuoles of some macrophages; other macrophages contained intracytoplasmic aggregates of calcified coccobacilli.
Specific Detection of Shedding and Latency of Bovine Herpesvirus 1 and 5 using a Nested Polymerase Chain Reaction
Authors: Ashbaugh Scott E. , Thompson Karin E. , Belknap Ellen B. , Schultheiss Patricia C. , Chowdhury Shafiqul , Collins James K.
Published: May 2011

A sensitive method for simultaneously detecting and discriminating between bovine herpesviruses types 1 and 5 (BHV-1 and BHV-5) was developed using a nested polymerase chain reaction (PCR) technique. Following amplification using type-common primers derived from gC sequences, amplification using type-specific nesting primers produced different-sized bands specific to the corresponding types, as demonstrated by blot hybridization. Less than 0.1 plaque-forming units (PFU) of each virus and 75 fg or less of viral DNA were routinely detected. The PCR technique amplified correct product from 4 BHV-5 isolates and from 48 BHV-1 isolates, all from the United States, and did not amplify heterologous herpesviruses. The PCR technique was more sensitive than virus isolation in detection of BHV-1 or BHV-5 in nasal secretions from experimentally and naturally infected calves, and it detected BHV-1 or BHV-5 in trigeminal ganglia from these calves.
Isolation of Porcine Circovirus-like Viruses from Pigs with a Wasting Disease in the USA and Europe
Authors: Allan G. M. , McNeilly F. , Kennedy S. , Daft B. , Ellis J. A. , Haines D. M. , Meehan B. M. , Adair B. M.
Published: May 2011

Samples of lung, liver, kidney, pancreas, spleen, and lymph node from pigs with postweaning multisystemic wasting syndrome from California (USA) and samples of mesenteric lymph nodes from similarly diseased pigs from Brittany (France) were examined by light microscopy, in situ hybridization (ISH), and/or virus isolation. Whole genomic probes for porcine circovirus (PCV) and chicken anemia virus (CAV) were used for ISH. Tissue homogenate supernatants were inoculated onto PK/15 cells for virus isolation, and the presence of viral antigen and viral particles was verified by indirect immunofluorescence, ISH, and electron microscopy. Histologic examination of lung from pigs from California revealed interstitial pneumonia, alveolar epithelial hyperplasia, and basophilic nuclear and cytoplasmic inclusions in mononuclear cell infiltrates and various pulmonary epithelial cells. Granulomatous lymphadenitis with syncytial cells typified the lesions seen in the pigs from France. PCV-like nucleic acid was detected by ISH in lung, pancreas, lymph node, kidney, and liver in pigs from California. Positive signal was also obtained in lymph node sections from pigs from France. Probes for CAV were consistently negative. PK/15 cell cultures inoculated with lung preparations from diseased California pigs and mesenteric lymph node preparations from pigs from France had positive fluorescence by indirect staining for PCV using pooled polyclonal pig sera and hyperimmune rabbit serum and had variable staining with a panel of 7 monoclonal antibodies specific for cell culture contaminant PCV. PCV-like nucleic acid was also detected by ISH in cell cultures. Cytopathic effect was not observed Electron microscopic examination of inoculated cell cultures revealed 17-nm viral particles morphologically consistent with PCV No other virus particles were observed. Although genomic analysis for the definitive identification of these viral isolates remains to be done, the evidence provided strongly suggests that these tissue isolates are closely related to, although antigenically distinct from, the original PCV cell culture contaminant.
Risk Analysis of Quarantine Station Performance: A Case Study of the Importation of Equine Infectious Anemia Virus-Infected Horses into California
Authors: Carpenter Tim E. , McBride Michael D. , Hird David W.
Published: May 2011

We examined the risk of importing and mistakenly releasing equine infectious anemia virus (EIAV)-infected horses into California. A computer simulation model was constructed to evaluate current and alternative quarantine station procedures; 150,000 iterations were performed to simulate 15 different scenarios of 10,000 horses imported into the state over a 14-year period. Simulation results showed that under current conditions of low EIAV prevalence in exporting countries, increasing the quarantine period would not decrease the number of EIAV-infected horses mistakenly released from quarantine. In a worst case scenario of high EIAV prevalence in exporting countries, the model predicted 10 EIAV-infected horses would be imported, of these 1 or none would escape detection and would be released mistakenly if quarantine duration were 3 or 14 days, respectively. This model may be applied to other quarantine station situations for evaluating the importation risk for EIAV and other diseases.
Changes in Levels of Viremia in Cattle Persistently Infected with Bovine Viral Diarrhea Virus
Authors: Brock K. V. , Grooms D. L. , Ridpath J. , Bolin S. R.
Published: May 2011

Virus isolation and serum neutralizing antibody titers were determined over a period of time from samples collected from animals persistently infected with bovine viral diarrhea virus (BVDV). To evaluate over time the ability to detect BVDV by virus isolation from serum or white blood cell preparations, 4 persistently infected calves were monitored from birth until 70 days of age. In 3 of 4 persistently infected calves, virus isolation from serum and white blood cells was negative until approximately 42 days of age, when colostral antibody had declined. The level of viremia in 7 adult (>12 months) persistently infected animals decreased by 1 10-fold dilution over at least a 2-year period. The level of viremia became undetectable by virus isolation from serum in 1 of the 7 animals examined. This decline was associated with the development of virus neutralizing antibody. Although the level of viremia is fairly stable within persistently infected animals, the presence of specific neutralizing antibody may affect the ability to isolate BVDV. These findings are important when considering diagnostic testing to identify persistently infected animals by virus isolation.
Severe Acute Bovine Viral Diarrhea in Ontario, 1993–1995
Authors: Carman Susy , van Dreumel Tony , Ridpath Julia , Hazlett Murray , Alves David , Dubovi Ed , Tremblay Rob , Bolin Steve , Godkin Ann , Anderson Neil
Published: May 2011

In 1993, noncytopathic bovine viral diarrhea virus (BVDV) strains with enhanced virulence caused unprecedented outbreaks of severe acute bovine viral diarrhea (BVD) in dairy, beef, and veal herds in Ontario (Canada). Fever, pneumonia, diarrhea, and sudden death occurred in all age groups of cattle. Abortions often occurred in pregnant animals. Gross lesions in the alimentary tract were similar to those associated with mucosal disease, especially in animals >6 months of age. Cattle of all age groups had microscopic lesions in the alimentary tract similar to those seen with mucosal disease. The epidemic peaked in the summer of 1993, with 15% of all bovine accessions from diseased cattle presented to the diagnostic laboratory being associated with BVDV. The virus strains involved in the outbreak were analyzed using monoclonal and polyclonal antibodies and the polymerase chain reaction. The virus isolates from these outbreaks of severe disease were determined to be type 2 BVDV. Type 2 BVDV has been present in Ontario at least since 1981 without causing widespread outbreaks of severe acute BVD, which suggests that type 2 designation in itself does not imply enhanced virulence. Cattle properly vaccinated with type 1 BVDV vaccines appear to be protected from clinical disease.
Enteric Lesions Induced by Different Pseudorabies (Aujeszky's Disease) Virus Strains Inoculated into Closed Intestinal Loops of Pigs
Authors: Narita M. , Zhao Y. M. , Kawashima K. , Arai S. , Hirose H. , Yamada S. , Ezura K.
Published: May 2011

Three different strains of pseudorabies (Aujeszky's disease) virus were inoculated into ligated closed loops in the jejunum and ileum of five specific-pathogen-free pigs. Infected areas were compared with respect to distribution of histologic lesions and pseudorabies virus antigen. Two wild-type strains of pseudorabies virus produced enteric lesions consisting of necrosis of the subepithelial macrophages in the basilar crypt epithelium, necrosis of the lymphoid follicles in the Peyer's patches, degeneration of the epithelial cells in the crypt and villi, degeneration of the neuronal cells in the myenteric plexuses, and formation of intranuclear inclusion bodies on postinoculation days 2–4. Pseudorabies virus antigen was initially detected in subepithelial macrophages of the dome of Peyer's patches on postinoculation day 2 and subsequently extended to superficial epithelium and deeper into the lymphoid follicles and myenteric plexuses on postinoculation days 3–4. Many pseudorabies virus particles were also detected in the center of their necrotic foci. However, 1 mutant strain (ara-T-resistant) of pseudorabies virus did not produce enteric lesions. The results suggest that the primary target of infection by wild-type strains of pseudorabies virus might be the macrophages distributed in the subepithelial area of the dome Peyer's patches.
Evaluation of a Single Dilution ELISA System for Detection of Seroconversion to Bovine Viral Diarrhea Virus, Bovine Respiratory Syncytial Virus, Parainfluenza-3 Virus, and Infectious Bovine Rhinotracheitis Virus: Comparison with Testing by Virus Neutralization and Hemagglutination Inhibition
Authors: Graham D. A. , McShane J. , Mawhinney K. A. , McLaren I. E. , Adair B. M. , Merza M.
Published: May 2011

A single-dilution quantitative enzyme-linked immunosorbent assay (ELISA) system, based on commercial ELISA kits, for the simultaneous detection of seroconversion to bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), parainfluenza-3 virus (PI3V), and infectious bovine rhinotracheitis virus (IBRV) was evaluated by testing acute and convalescent serum pairs from 564 cattle in 145 outbreaks of respiratory disease. Seroconversion to BVDV, BRSV, PI3V and IBRV was detected in 8.0%, 19.0%, 13.7%, and 7.4%, respectively, of serum pairs tested. Seroconversion was detected in 60.7% of herds and 34.6% of animals tested. Infection with 2 or more viruses was found in 46.6% of these herds and in 27.2% of these animals. The majority of BVDV infections (62%) were associated with other virus infections, suggesting that BVDV may potentiate infection with other agents rather than being a primary pathogen of the respiratory tract. The results were compared with those obtained by virus neutralization and hemagglutination inhibition testing, and the sensitivity, specificity, and overall correlation were calculated. Sensitivities of 92%, 95%, 100%, and 100% were obtained for BVDV, BRSV, PI3V, and IBRV, respectively. The corresponding specificity values were 89%, 92%, 86%, and 91%. The overall correlation for each virus was 90%, 93%, 90%, and 93%, respectively. These results demonstrate that this ELISA system may be used successfully to detect seroconversion in serum pairs, highlight the frequency of multiple viral infections in outbreaks of respiratory disease, and provide further evidence of an immunosuppressive role for BVDV infections.
Detection of Human Granulocytic Ehrlichiosis Agent and Borrelia Burgdorferi in Ticks by Polymerase Chain Reaction
Authors: Chang Yung-Fu , Novosel Vesna , Chang Chao-Fu , Kim Jong Bae , Shin Sang J. , Lein Donald H.
Published: May 2011

Adult ixodid ticks were collected from Westchester County, New York, and Ipswich, Massachusetts, to determine the presence of infection with a human granulocytic ehrlichiosis (HGE) agent by using the polymerase chain reaction (PCR). The presence of Borrelia burgdorferi in ticks collected from New York was also determined by PCR. Of the 229 ticks from New York and 47 ticks from Massachusetts, 9% (22/229) and 25% (12/47) of ticks contained HGE agent, respectively. Fifty-four percent (123/229) of the ticks collected from New York were B. burgdorferi positive; 4% (9/229) of these ticks contained both HGE agent and B. burgdorferi. This finding indicates that animals with Lyme borreliosis may be also exposed to the etiologic agent of HGE. More extensive laboratory diagnosis may be necessary when multiple tick-borne diseases are suspected in animals.
Detection of Cytopathic Bovine Viral Diarrhea Virus in the Ovaries of Cattle following Immunization with a Modified Live Bovine Viral Diarrhea Virus Vaccine
Authors: Grooms Daniel L. , Brock Kenny V. , Ward Lucy A.
Published: May 2011

Economic loss from infection with bovine viral diarrhea virus (BVDV) is of worldwide concern. The unique pathogenesis and antigenic variability of BVDV have made this virus challenging to control. Vaccination programs are a major component of control and prevention strategies. Both killed and modified live vaccines are commercially available. Choice between killed and modified live vaccines is controversial. Of major concern is the safety of modified live vaccines. Little information is available on their tissue tropism and potential for causing pathology, especially with respect to the reproductive system. The objective of this study was to determine if BVDV could be detected in the ovary of cattle following immunization with a modified live BVDV vaccine. In 2 separate trials, 6 heifers and 4 mature cows were immunized with a modified live BVDV vaccine and ovaries were removed between 7 and 30 days postvaccination. Cytopathic BVDV was isolated from ovaries removed on days 8, 10, and 12. BVDV antigen was detected using immunohistochemistry on days 10–30. These findings are significant because replication of virus in the ovary could cause ovarian dysfunction, resulting in reduced fertility.
Prevalence of Bovine Viral Diarrhea Virus Genotypes and Antibody against those Viral Genotypes in Fetal Bovine Serum
Authors: Bolin Steven R. , Ridpath Julia F.
Published: May 2011

One thousand lots of pooled fetal bovine serum (FBS) were tested for contamination with bovine viral diarrhea virus (BVDV) and/or for contamination with neutralizing antibody against BVDV. Noncytopathic or cytopathic BVDV was isolated from 203 lots of FBS. Analysis of the viral isolates identified 115 type 1 and 65 type 2 BVDV isolates. An additional 23 virus isolates were mixtures of >2 BVDV isolates and were not classified to viral genotype. Further characterization of the type 1 viruses identified 51 subgenotype 1a and 64 subgenotype 1b BVDV isolates. Viral neutralizing antibody was detected in 113 lots of FBS. Differential viral neutralization indicated that type 1 BVDV induced the antibody detected in 48 lots of FBS and type 2 BVDV induced the antibody detected in 16 lots of FBS.
Differentiation of a Porcine Reproductive and Respiratory Syndrome Virus Vaccine Strain from North American Field Strains by Restriction Fragment Length Polymorphism Analysis of ORF 5
Authors: Wesley Ronald D. , Mengeling William L. , Lager Kelly M. , Clouser Deborah F. , Landgraf John G. , Frey Merwin L.
Published: May 2011

The suitability of restriction fragment length polymorphism (RFLP) analysis for differentiating a porcine reproductive and respiratory syndrome virus (PRRSV) vaccine strain from other North American field strains was investigated. Open reading frame 5 nucleotide sequence data of the vaccine virus, its parent strain VR-2332, and 22 other strains of PRRSV included in this study indicated that 3 restriction enzyme gel patterns characterize the vaccine virus and the parent strain genotype. The combined 3 RFLP patterns differentiate the vaccine and parent virus from other PRRSV strains. This test will be a valuable tool in epidemiologic studies and will be useful in identifying individual strains in cases of multistrain PRRSV infections.
Gastrointestinal Pathogenicity of Adenoviruses and Reoviruses Isolated from Broiler Chickens in Alabama
Authors: Lenz Stephen D. , Hoerr Frederic J. , Ellis Alfred C. , Toivio-Kinnucan Maria A. , Yu Maria
Published: May 2011

Adenoviruses and reoviruses isolated from commercial broiler chickens were evaluated for gastrointestinal pathogenicity in specific-pathogen-free Leghorn chickens. The viruses were originally isolated from either the proventriculus or a gastrointestinal pool of tissues of broiler chickens with proventriculitis or enteritis. Isolates were cloned by terminal dilution. Day-old chickens were inoculated by oral and ocular routes with undiluted tissue culture fluids (titers of 102-104 TCID50/ml) and then examined at necropsy on days 5, 10, and 15 postinoculation. Chickens in all virus groups (but not the control group) developed wet, unformed fecal droppings that persisted for the duration of the study. Mild lesions occurred in reovirus-inoculated chickens and included hyperplasia of lymphocyte aggregates in various organs and mild gizzard erosions. Chickens inoculated with adenovirus isolates developed marked gizzard erosions and necrotizing pancreatitis as well as mild proventriculitis. Intranuclear viral inclusion bodies occurred in gizzard epithelium and pancreatic acinar cells at the sites of lesions. Lymphocytic atrophy occurred in the bursa of Fabricius. Respective viruses were reisolated from proventriculus and duodenum collected from chickens of each group; no viruses were isolated from controls. Under the conditions of this study, adenovirus isolates were more pathogenic than the reovirus isolates in the digestive system.
Evaluation of a Computerized Antimicrobial Susceptibility System with Bacteria Isolated from Animals
Authors: Hubert Susannah K. , Nguyen Phouc Dinh , Walker Robert D.
Published: May 2011

The BIOMIC is a computerized system used to calculate the minimal inhibitory concentration (MIC) of an antimicrobial agent from a zone of inhibition generated by a disk diffusion test. This system was developed using bacterial pathogens of human origin. This study investigated the use of the BIOMIC system for determining MICs for bacterial pathogens from animals. The MICs generated by the BIOMIC system were compared with the MICs generated using a broth microdilution testing method. A total of 663 drug—organism combinations was tested. These combinations included 3 species of gram-positive bacteria, 5 species of gram-negative bacteria, and the antimicrobial agents ampicillin, gentamicin, cephalothin, ciprofloxacin, enrofloxacin, trimethoprim/sulfamethoxazole, tetracycline, and erythromycin. Overall, the MICs generated by the BIOMIC system correlated with the broth microdilution MICs for 72% of the total drug—organism combinations tested. The Pseudomonas aeruginosa strains tested showed the highest agreement between the 2 systems, with 100% for all antibacterial agents tested, whereas Pasteurella haemolytica, Pasteurella multocida, and enterococci showed the least agreement (76%, 57%, and 47%, respectively). Among these organisms, trimethoprim-sulfa showed the least agreement (31%) and ciprofloxacin showed the greatest (91%). These results indicate that the BIOMIC system could be a useful tool in veterinary medicine for producing quantitative antimicrobial susceptibility results. However, it is currently unreliable for some drug—bacteria combinations. This discrepancy possibly could be corrected by modification of the software using data points generated by a large-scale study.
Normal Reference Intervals and the Effects of Time and Feeding on Serum Bile Acid Concentrations in Llamas
Authors: Andreasen Claire B. , Pearson Erwin G. , Smith Brad B. , Gerros Terry C. , Lassen E. Duane
Published: May 2011

Fifty clinically healthy llamas, 0.5–13 years of age (22 intact males, 10 neutered males, 18 females), with no biochemical evidence of liver disease or hematologic abnormalities, were selected to establish serum bile acid reference intervals. Serum samples submitted to the clinical pathology laboratory were analyzed using a colorimetric enzymatic assay to establish bile acid reference intervals. A nonparametric distribution of llama bile acid concentrations was 1–23 μmol/liter for llamas >1 year of age and 10–44 μmol/liter for llamas ≤1 year of age. A significant difference was found between these 2 age groups. No correlation was detected between gender and bile acid concentrations. The reference intervals were 1.1–22.9 μmol/liter for llamas >1 year of age and 1.8–49.8 μmol/liter for llamas ≤1 year of age. Additionally, a separate group of 10 healthy adult llamas (5 males, 5 females, 5–11 years of age) without biochemical or hematologic abnormalities was selected to assess the effects of feeding and time intervals on serum bile acid concentrations. These 10 llamas were provided fresh water and hay ad libitum, and serum samples were obtained via an indwelling jugular catheter hourly for 11 hours. Llamas were then kept from food overnight (12 hours), and subsequent samples were taken prior to feeding (fasting baseline time, 23 hours after trial initiation) and postprandially at 0.5, 1, 2, 4, and 8 hours. In feeding trials, there was no consistent interaction between bile acid concentrations and time, feeding, or 12-hour fasting. Prior feeding or time of day did not result in serum bile acid concentrations outside the reference interval, but concentrations from individual llamas varied within this interval over time.
Prior Infection of Nursery-Age Pigs with Porcine Reproductive and Respiratory Syndrome Virus Does not Affect the Outcome of Transmissible Gastroenteritis Virus Challenge
Authors: Wesley Ronald D. , Mengeling William L. , Lager Kelly M.
Published: May 2011

Thirty-six specific-pathogen-free pigs were weaned at 2 weeks of age and separated into 4 treatment groups (A—D, 9 pigs/group). Treatment groups B and D were infected with porcine reproductive and respiratory syndrome virus (PRRSV), whereas groups A and C remained uninfected. Two weeks later, 1 pig from each group was necropsied to assess gross lung involvement, and then the remaining group D PRRSV-infected pigs and the group C uninfected pigs were challenged at 4 weeks of age with transmissible gastroenteritis virus (TGEV) to determine if prior infection with PRRSV increased the severity of TGEV disease after challenge. One hundred percent morbidity but no mortality occurred in pigs following challenge. Clinically, pigs of both groups C and D were similar in terms of onset and severity of diarrhea. The serum antibody response to TGEV and the amount and duration of TGEV shedding after challenge was similar for both groups. Only a few pigs in each group had a transient fever postchallenge, and both group C and group D pigs began to recover and to gain weight at or near the end of the first week postchallenge. It was concluded that the clinical course of TGEV disease was not markedly affected by infection of pigs with TGEV 2 weeks after they had been infected with PRRSV.
Immunohistochemical Detection of Ki-67 and PCNA in Canine Mammary Tumors: Relationship to Clinical and Pathologic Variables
Authors: Peña Laura L. , Nieto Ana I. , Pérez-Alenza Dolores , Cuesta Pedro , Castaño Maria
Published: May 2011

The objectives of this study were to measure the proliferation indices in canine mammary tumors using immunohistochemical detection of Ki-67 and proliferating cell nuclear antigen (PCNA), to determine the relationship of these antigens to clinical and pathologic variables, and to investigate the usefulness of these antigens as prognostic indicators. Ninety-six female dogs with 115 primary nonmetastasized spontaneous mammary tumors and dysplasias were included in the study. Immunostaining was performed using MIB-1 and PC10 monoclonal antibodies against Ki-67 and PCNA, respectively. Ki-67 and PCNA proliferation indices were determined. Dogs were followed for 18 months, with clinical examinations every 3–4 months. There was a significant correlation between Ki-67 and PCNA indices in the dogs with dysplasias and benign tumors but not in the dogs with malignant tumors. The clinical stage at first presentation was related to the proliferative index measured with Ki-67 but not to that measured with PCNA. Proliferation indices were significantly lower in the nonmalignant tumors and dysplasias than in the malignant tumors. In malignant tumors, the PCNA index had a positive correlation with the histologic malignant grade and the nuclear grade. High index values of Ki-67 were positively correlated with metastasis, death from neoplasia, low disease-free survival rates, and low overall survival rates. PCNA displayed no significant association with these variables. Multivariate analyses concerning metastasis, disease-free survival, and overall survival revealed that the Ki-67 index had prognostic value.
Malignant Catarrhal Fever in Bison, Acute and Chronic Cases
Authors: Schultheiss Patricia C. , Collins James K. , Austgen Laura E. , DeMartini James C.
Published: May 2011

Acute malignant catarrhal fever (MCF) was diagnosed in 10 bison from 6 herds and ranging from 1 to 6 years of age. The pattern of clinical signs and morphologic lesions differed among bison. Combinations of corneal opacity, lacrimation, nasal discharge, depression, excess salivation, anorexia, diarrhea, melena, and hematuria were observed. Vasculitis characterized by lymphoid infiltrates in the adventia with variable extension into media and intima was found in multiple tissues in each animal. Fibrinoid vascular necrosis was rare. Ulceration in the alimentary tract was found in 9/10 bison, and ulceration or hemorrhage in the urinary bladder was found in 8/10 bison. Lymphoid infiltrates were present in 7 of 9 livers and 9 of 9 kidneys examined histologically. Hyperplasia of lymph nodes was observed in 5 bison. Chronic MCF was diagnosed in 1 bison with an 80-day course of illness that began with lacrimation, corneal opacity, mucoid nasal discharge, depression, and anorexia. These signs ceased after 15 days but circling and blindness developed on day 76. Chronic vascular lesions characterized by endothelial cell hypertrophy, intimal thickening, fragmentation of the internal elastic membrane, smooth muscle hypertrophy, and adventitial infiltrates of lymphocytes and plasma cells were found in many organs. The retinal arteries had chronic inflammation and acute transmural fibrinoid necrosis. The retinas were infarcted. Polymerase chain reaction technique for amplification of ovine herpesvirus 2 sequences was performed on formalin-fixed tissues, and viral sequences were detected in 1–7 tissues from each animal. These viral sequences were not found in tissues of 4 bison not affected by MCF.
Enzyme-Linked Immunosorbent Assay for Detection of Feline Herpesvirus 1 IgG in Serum, Aqueous Humor, and Cerebrospinal Fluid
Authors: Dawson D. A. , Carman J. , Collins J. , Hill S. , Lappin M. R.
Published: May 2011

Feline herpesvirus 1 (FHV) is an important cause of feline ocular and respiratory disease, but the role the virus plays in central nervous system disease of cats has not been explored. The study described here was performed to validate an indirect enzyme-linked immunosorbent assay (ELISA) to detect FHV-specific IgG antibodies for use in feline epidemiologic, ocular, and central nervous system disease studies. The indirect IgG ELISA was applied to serum, aqueous humor, and cerebrospinal fluid from cats. Serum FHV IgG ELISA results were compared with those of serum neutralization in client-owned cats and laboratory-housed cats following vaccination. Of the 100 client-owned cats tested by ELISA, 97 had detectable FHV IgG; 95 had titers >32. The FHV IgG ELISA was more sensitive than serum neutralization and could be used with aqueous humor and cerebrospinal fluid. Cats with inflammatory central nervous system or ocular diseases had significant leakage of serum proteins into aqueous humor and cerebrospinal fluid, necessitating use of cutoff values derived from serum when these fluids were assessed.
Use of Polymerase Chain Reaction and Immunohistochemistry for Detection of Canine Adenovirus Type 1 in Formalin-Fixed, Paraffin-Embedded Liver of Dogs with Chronic Hepatitis or Cirrhosis
Authors: Chouinard Luc , Martineau Daniel , Forget Céline , Girard Christiane
Published: May 2011

To assess the possible involvement of canine adenovirus type 1 (CAV-1) in naturally occurring cases of canine chronic liver disease, a polymerase chain reaction (PCR)-based assay was developed to detect a conserved region of the major core protein gene ( pVII) of CAV-1 in formalin-fixed, paraffin-embedded liver sections. Results were compared with a standard avidin-biotin immunoperoxidase complex technique that detected CAV-1 antigens using a commercial monoclonal anti-adenovirus antibody. Seventeen cases of cirrhosis and 28 cases of chronic hepatitis with piecemeal necrosis and progressive fibrosis were selected for the study. Formalin-fixed, paraffin-embedded liver sections of 2 cases of infectious canine hepatitis (ICH) and crude DNA extract from CAV-1 (ATCC VR 293 Utrecht strain) served as positive controls. A 411-base-pair viral region was amplified and sequenced as CAV-1 pVII in both cases of infectious canine hepatitis and in the CAV-1 crude DNA extract. The 2 ICH cases were positive for CAV-1 antigens by the immunoperoxidase method. CAV-1 DNA or antigens were not detected by either technique in any of the 45 cases of chronic liver disease selected for the study. These results indicate that both PCR and immunohistochemistry are reliable and rapid techniques for detecting CAV-1 in formalin-fixed, paraffin-embedded liver sections of dogs with ICH. Several possibilities may explain the negative results obtained with both techniques in this study, including the noninvolvement of CAV-1 in canine chronic hepatitis and cirrhosis and the possibility that the virus causes initial damage, provokes a self-perpetuating chronic liver disease, and disappears.
Seasonal Values of Selected Blood Parameters of Farm-Raised Channel Catfish (Ictalurus Punctatus) in the Mississippi Delta
Authors: Leard A. Timothy , Wagner Bruce A. , Camp Karen L. , Wise David J. , Gao Xio Dan
Published: May 2011

Hematocrit, sodium, chloride, potassium, calcium, glucose, and pH were measured in whole blood of 1,522 channel catfish collected from 3 commercial food-fish ponds in the Mississippi Delta. Samples were collected from March 1995 to March 1996 to monitor seasonal fluctuations. A total of 10–20 fish were arbitrarily collected with snag lines from each pond on each sample day. The mean monthly hematocrits fluctuated seasonally from a low of 14.5% in midwinter to a high of 25.7% in midsummer (annual = 21%, SE = 0.15). Sodium levels were consistent throughout the year with a mean (SE) of 134 (0.13) mM/liter. Mean chloride values for the year were 120 (0.14) mM/liter but increased to 132 mM/liter in midwinter. By March 1996, the chloride levels had returned to levels observed during spring 1995. Potassium and glucose levels varied throughout the year with means of 4.43 (0.06) mM/liter and 26.9 (0.46) mg/dl, respectively, and coefficients of variation of 51.8% and 63.3%, respectively. Calcium and pH values were fairly stable with means of 1.31 (0.004) mM/liter and 7.13 (0.004), respectively. All parameters except glucose and potassium may be adequately evaluated with a sample size of 25 or less. These data were collected to provide baseline information for ongoing pond health studies.
Reproduction of Lesions of Postweaning Multisystemic Wasting Syndrome in Gnotobiotic Piglets
Authors: Ellis John , Krakowka Steven , Lairmore Michael , Haines Deborah , Bratanich Ana , Clark Edward , Allan Gordon , Konoby Carrie , Hassard Lori , Meehan Brian , Martin Karen , Harding John , Kennedy Seamus , McNeilly Francis
Published: May 2011

Neonatal gnotobiotic piglets were inoculated with tissue homogenates and low- and high-passage cell culture material to determine if the lesions of the newly described porcine postweaning multi-systemic wasting syndrome (PMWS) could be reproduced. For this, 17 3-day-old gnotobiotic piglets were inoculated intranasally with pelleted chloroform-treated, filtered extracts from cell cultures, filter-sterilized homogenates of lymphoid tissue from PMWS-affected piglets, or control materials. Piglets were maintained in germ-free isolators for up to 5 weeks after infection prior to euthanasia and collection of samples for analysis. All piglets inoculated with the viral inocula developed lesions typical of PMWS, including generalized lymphadenopathy, hepatitis, nephritis, interstitial pneumonia, myocarditis, and gastritis. Porcine circovirus (PCV), as well as porcine parvovirus (PPV), was detected in tissues by virus reisolation, polymerase chain reaction analysis, or immunohistochemistry. All infected piglets developed moderate to high titers of antibody to PCV and moderate titers to PPV. No lesions, virus, or virus-specific antibodies were detected in sham-inoculated or uninoculated control piglets. These studies demonstrate that the lesions of PMWS can be experimentally reproduced in gnotobiotic piglets using filterable viral agents derived from pigs with PMWS and provide an experimental basis for further investigation into the pathogenesis and control of this emerging infectious disease in swine.
Sensitivity Comparison for Detection of Respiratory Bovine Coronaviruses in Nasal Samples from Feedlot Cattle by ELISA and Isolation with the G Clone of HRT-18 Cells
Authors: da Silva Manuel Reis , O'Reilly Kathy L. , Lin Xiaoqing , Stine Lisa , Storz Johannes
Published: May 2011

A monoclonal antibody-based capture enzyme-linked immunosorbent assay (ELISA) was developed to detect respiratory bovine coronavirus (RBCV) antigens in nasal swabs collected from cattle showing signs of respiratory tract disease following shipping. These samples had been previously tested for RBCV by inoculation of G clone cultures of human rectal tumor cells (HRT-18G) and for bovine herpes virus 1, parainfluenza virus 3, bovine adenovirus, bovine respiratory syncytial virus, and bovine viral diarrhea virus on other specifically permissive cell cultures. RBCV has not previously been recognized as an important etiological factor in the bovine respiratory disease complex of feedlot cattle. Thirty of 100 samples tested positive for RBCV antigen by capture ELISA in contrast to 38 of 100 samples that yielded RBCV isolates in G clone cells. Samples yielding other bovine respiratory viruses in the absence of RBCV were negative in the capture ELISA, which was based on the use of a single monoclonal antibody that recognizes one RBCV epitope on the S glycoprotein with the broadest reactivity with different strains of RBCV tested. Some RBCV strains may not be detected by this ELISA, which may account for the higher percentage of RBCV-infected cattle detected by RBCV isolation. However, the ELISA was simple to perform, sensitive, and specific and was more rapid than virus isolation. This assay will be useful for processing large numbers of field samples in future epidemiologic and diagnostic studies of RBCV infections of cattle.
Detection of Porcine Reproductive and Respiratory Syndrome Virus by Reverse Transcription-Polymerase Chain Reaction using Different Regions of the Viral Genome
Authors: Guarino Helena , Goyal Sagar M. , Murtaugh Michael P. , Morrison Robert B. , Kapur Vivek
Published: May 2011

Serologic studies have revealed strain variability between American and European isolates and among American isolates of porcine reproductive and respiratory syndrome virus (PRRSV). The objective of this study was to develop an assay for the routine diagnosis of PRRSV in field specimens using reverse transcription-polymerase chain reaction (RT-PCR) amplification of conserved genomic regions. Twenty-four field isolates of PRRSV from different regions of the USA were analyzed in the study. Six primer pairs from open reading frames (ORFs) 4, 6, and 7 of the American strain (ATCC VR-2332) and from ORF 1b of the Lelystad strain were used for the amplification of the viral genome by PCR. Amplification products of the expected sizes were obtained from all isolates by PCR amplification of ORF 7, the gene encoding the nucleocapsid protein. Oligonucleotide primers designed to amplify ORFs 4 and 6 detected 92% and 96% of the isolates, respectively, whereas primers for the amplification of ORF 1b detected 88% of all isolates. The specificity of the amplified products of ORF 7 from 7 field isolates and 2 reference strains was confirmed by chemiluminescent hybridization using an internal digoxigenin-labeled DNA probe. Sequence analysis of this region indicated variation in the nucleotide sequence of 2 isolates that did not hybridize with the internal probe. These results indicate that ORF 7 may serve as a potential target for the detection of PRRSV strains by RT-PCR and that genomic variability should be considered when nucleic acid hybridization is used to confirm the specificity of PCR amplification for diagnostic purposes.
Differential Serologic Response to Mycoplasma Ovipneumoniae and Mycoplasma Arginini in Lambs Affected with Chronic Respiratory Disease
Authors: Niang Mamadou , Rosenbusch Ricardo F. , Lopez-Virella Jose , Kaeberle Merlin L.
Published: May 2011

An enzyme-linked immunoabsorbent assay (ELISA) was used to evaluate the levels of antibodies to Mycoplasma ovipneumoniae and M. arginini in lambs with chronic respiratory disease. Sera were obtained from lambs in several flocks at various stages of the clinical disease and tested with sodium dodecyl sulfate (SDS)-treated M. ovipneumoniae and M. arginini whole cells and a crude capsular extract of M. ovipneumoniae as the antigens. There were low levels of antibody to M. ovipneumoniae in flocks sampled at the early stages of infection, whereas increased levels of antibody were present in lambs from flocks that had apparently recovered from the clinical disease. Slowly rising titers of circulating antibodies to M. ovipneumoniae were confirmed by sequential bleeding of lambs during the course of the clinical disease. However, antibody levels of M. arginini were more likely to increase earlier in the disease process. There was significant cross-reactivity between the 2 SDS-treated antigens in both the ELISA test and western immunoblotting. In contrast, the crude capsular extract was specific for detecting antibodies to M. ovipneumoniae.
An IgG Avidity ELISA to Discriminate between Recent and Chronic Neospora Caninum Infection
Authors: Björkman Camilla , Näslund Katarina , Stenlund Susanne , Maley Stephen W. , Buxton David , Uggla Arvid
Published: May 2011

The avidity of IgG antibodies directed to Neospora caninum was measured using an IgG avidity enzyme-linked immunosorbent assay (ELISA) employing N. caninum proteins incorporated into immunostimulating complexes as antigen. In this ELISA, low-affinity antibodies were eluted by adding an incubation step with urea after the serum incubation. The antibody titers obtained with and without incubation with urea were then used to calculate the IgG avidity values. Analysis of sequential sera collected from experimentally infected calves revealed that the avidity increased during the course of infection. Three weeks after infection, the IgG avidity was 9–18%, and 24 weeks later it had increased to 58–76%. Cattle naturally infected for more than 6 months all had avidities >50%. The results in this study, however preliminary, indicate that the IgG avidity ELISA can be used to discriminate between recent and chronic N. caninum infections and may therefore be a valuable complement to IgG assays in epidemiologic studies of N. caninum infection in cattle.