bluuebunny/crossref_metadata_2025 huggingface dataset:
Congenital hydranencephaly and cerebellar hypoplasia in water buffalo in southern Brazil
Authors: Schild Ana Lucia , Fiss Letícia , Damé Maria C. , Uzal Francisco A. , Soares Mauro P. , Schuch Luiz F. , Flores Eduardo F. , Riet-Correa Franklin
Published: May 2011

Seven cases of hydranencephaly and cerebellar hypoplasia in a water buffalo herd are described. The herd of 133 females was divided for mating into 4 groups and mated with different bulls. The disease was observed in the offspring of 5 cows from only 1 group over a period of 6 years. These cows had all mated with the same bull during that period. All affected calves were unable to suckle, showed intention tremors, involuntary movements of the ears, depression, and blindness. Some calves were recumbent and others showed wide-based stance and inability to walk. At necropsy, all affected calves had similar lesions consisting of hydranencephaly and cerebellar hypoplasia. Histologically, the cavities were surrounded by normal-looking nervous tissue. The occurrence of the disease in the offspring of only 1 out of 6 bulls in different years, the similar lesions in all 7 calves affected, and the negative serology and immunohistochemistry for Bovine virus diarrhea virus and Bluetongue virus highly suggest that the disease is hereditary.
Development of a reverse transcription loop-mediated isothermal amplification assay for detection of Porcine teschovirus
Authors: Wang Bin , Wang Yongqiang , Tian Zhi-Jun , An Tong-Qing , Peng Jin-Mei , Tong Guang-Zhi
Published: May 2011

Loop-mediated isothermal ampliﬁcation (LAMP) is a sensitive method for DNA ampliﬁcation. In the present report, the development of a single-tube, one-step, real-time accelerated reverse transcription (RT)-LAMP for the detection of Porcine teschovirus (PTV) is described. Six designed primers amplified target gene sequences successfully at constant temperature (65°C) within 1 hr, and the amplification results could be visualized directly by the naked eye. The sensitivity of the LAMP was 10 times higher than that of conventional polymerase chain reaction, and no cross-reactivity was found when the genomes of other common swine pathogens were subjected to the RT-LAMP system. When 43 clinical samples were tested by the RT-LAMP method, results indicated that the test is simple, rapid, accurate, and sensitive for the detection of PTV.
Baccharis megapotamica var. weirii poisoning in water buffalo (Bubalus bubalis)
Authors: Oliveira-Filho José C. , Carmo Priscila M. S. , Lucena Ricardo B. , Pierezan Felipe , Barros Claudio S. L.
Published: May 2011

An outbreak of an acute disease in buffalo ( Bubalus bubalis) caused by the ingestion of Baccharis megapotamica var. weirii occurred in the southern region of Brazil. Ten out of 50 buffalo died 24–48 hr after being introduced into a pasture containing abundant amounts of the plant. Factors influencing the ingestion of the plant and consequent toxicosis included hunger, stress caused by shipment, and unfamiliarity with the plant. Clinical signs included serous ocular discharge, incoordination, mild bloat, and muscle trembling. One buffalo was necropsied. Gross findings included dehydration, abundant liquid in the rumen, reddening of the mucosa of forestomachs, abomasum, and intestine, and edema of the wall of the rumen. The main histologic lesions were superficial to full thickness degeneration and necrosis of the stratified epithelium lining the forestomachs, necrosis of the intestinal mucosa, and widespread lymphoid necrosis. A calf ( Bos taurus) was fed a single dose of 5g/kg/body weight of B. megapotamica var. weirii harvested from the same site where the buffalo died. Twenty hours after the administration of the plant this calf died with clinical signs and lesions similar to those observed in the naturally poisoned buffalo.
Disseminated lipid-rich peritoneal mesothelioma in a horse
Authors: Dobromylskyj Melanie J. , Copas Victoria , Durham Andy , Hughes Thomas K. , Patterson-Kane Janet C.
Published: May 2011

A 9-year-old Haflinger mare presented to the Liphook Equine Hospital with a history of weight loss, azotemia, and repeated episodes of ascites over a period of 10 days. The horse was euthanized after exploratory laparotomy revealed large numbers of variably sized masses distributed throughout the peritoneal cavity. Macroscopically, some masses were papillary, while others were nodular. Histologically, the masses were comprised of large to giant, variably shaped, and occasionally multinucleated neoplastic cells with marked anisokaryosis and anisocytosis and a high mitotic rate. Small to moderate numbers of neoplastic cells were swollen by 1 to several, moderately sized to large, clear, circular or ovoid vacuoles, which stained positive with oil red O. Immunohistochemically, the neoplastic cells co-expressed vimentin and cytokeratin. Electron microscopy demonstrated tumor cells with tight junctions, microvilli, and numerous intracytoplasmic lipid droplets. These findings are consistent with a lipid-rich form of mesothelioma, which should be considered as a differential diagnosis if lipid vacuoles are present in potentially neoplastic cells in equine abdominocentesis samples.
Detection of a bacteriophage gene encoding a Mu-like portal protein in Haemophilus parasuis reference strains and field isolates by nested polymerase chain reaction
Authors: Zehr Emilie S. , Tabatabai Louisa B.
Published: May 2011

A nested polymerase chain reaction (nPCR) assay was developed to determine the presence of a gene encoding a bacteriophage Mu-like portal protein, gp29, in 15 reference strains and 31 field isolates of Haemophilus parasuis. Specific primers, based on the gene’s sequence, were utilized. A majority of the virulent reference strains and field isolates tested harbored the gene. The results suggest that the nPCR technique described in the current report could serve as a tool for epidemiological studies of H. parasuis.
Chondrosarcoma in a dromedary camel (Camelus dromedarius)
Authors: Janardhan Kyathanahalli S. , Ganta Chanran K. , Andrews Gordon A. , Anderson David E.
Published: May 2011

A 4-year 10-month-old, intact female dromedary camel had progressive left carpal joint swelling and lameness for 7 months. Radiographs showed multifocal lytic lesions in the carpal and proximal metacarpal bones. Surgical biopsy of the synovial capsule and carpal bones suggested neoplasia, and the camel was subsequently euthanized. At necropsy, a white to pale pink, firm, multilobulated, soft tissue mass was located on the palmar aspect of the left carpal joint. Two smaller masses were present on the dorsal aspect of the carpal joint. The masses infiltrated all the carpal bones and the proximal region of the metacarpal bone. The joint capsule was diffusely thickened. The articular surfaces of the carpal bones and the metacarpal bone were multifocally eroded. The lungs contained multiple, firm, raised, gray, randomly distributed nodules. The neoplastic cells stained positive for vimentin and S-100. Chondrosarcoma arising from around the carpal joint with infiltration of carpal and metacarpal bones, and pulmonary metastasis, was diagnosed based on the histopathological and immunohistochemical evaluation.
A new variant of Actinobacillus pleuropneumoniae serotype 3 lacking the entire apxII operon
Authors: Kuhnert Peter , Rohde Judith , Korczak Bożena M.
Published: May 2011

Actinobacillus pleuropneumoniae is an important respiratory pathogen causing pleuropneumonia in pig. The species is genetically characterized by the presence of 4 RTX ( Repeats in the Structural To Xin) toxin genes: apxI, apxII, and apxIII genes are differentially present in various combinations among the different serotypes, thereby defining pathogenicity; the apxIV gene is present in all serotypes. Polymerase chain reaction (PCR)-based apx gene typing is done in many veterinary diagnostic laboratories, especially reference laboratories. The present report describes the isolation of atypical A. pleuropneumoniae from 4 independent cases from 2 countries. All isolates were beta-nicotinamide adenine dinucleotide (β-NAD) dependent and nonhemolytic but showed strong co-hemolysis with the sphingomyelinase of Staphylococcus aureus on sheep blood agar. Classical biochemical tests as well as Matrix-assisted laser desorption ionization time-of-flight mass spectrometry and sequence-based analysis (16S ribosomal RNA [rRNA] and rpoB genes) identified them as A. pleuropneumoniae. Apx-toxin gene typing using 2 different PCR systems showed the presence of apxIV and only the apxIII operon ( apxIIICABD). None of the apxI or apxII genes were present as confirmed by Southern blot analysis. The 16S rRNA and rpoB gene analyses as well as serotype-specific PCR indicate that the isolates are variants of serotype 3. Strains harboring only apxIV and the apxIII operon are possibly emerging types of A. pleuropneumoniae and should therefore be carefully monitored for epidemiological reasons.
Development of a chromogenic in situ hybridization for Giardia duodenalis and its application in canine, feline, and porcine intestinal tissues samples
Authors: Weissenböck Herbert , Ondrovics Martina , Gurtner Susanne , Schiessl Peter , Mostegl Meike M. , Richter Barbara
Published: May 2011

In the present study, a chromogenic in situ hybridization for the identification of Giardia duodenalis in paraffin-embedded tissue samples was developed. The sensitivity and specificity of the probe was validated by testing it on cultured reference samples of different assemblages of G. duodenalis as well as culture and tissue samples containing other protozoa and infectious agents. The probe gave a positive reaction with the Giardia samples and a negative reaction with all other samples. Further, the probe was used for screening of histological slides of intestine from different animal species (99 canine samples, 85 feline samples, and 202 porcine samples) for the presence of G. duodenalis trophozoites. With this assay, the parasites were detected in samples from 8 dogs (8.08%), 6 cats (7.06%), and zero pigs. The results clearly indicate that the described method is useful for detection of Giardia trophozoites in routinely processed intestinal tissue of different animal species.
Development and analytical validation of a radioimmunoassay for the measurement of alpha1-proteinase inhibitor concentrations in feces from healthy puppies and adult dogs
Authors: Heilmann Romy M. , Paddock Casey G. , Ruhnke Isabelle , Berghoff Nora , Suchodolski Jan S. , Steiner Jörg M.
Published: May 2011

Canine α1-proteinase inhibitor (cα1-PI), a proteolysis-resistant protein with a molecular weight similar to albumin, has been shown to be clinically useful as a marker for gastrointestinal protein loss in dogs. A competitive, liquid-phase radioimmunoassay was developed and analytically validated. Fecal samples were collected from 101 healthy pet dogs of various breeds and ages, and fecal cα1-PI (Fcα1-PI) concentrations were compared between dogs of different age groups. A reference interval for Fcα1-PI concentration was calculated using the central 95th percentile. Analytical sensitivity of the assay was 2.2 µg Fcα1-PI/g feces. Observed-to-expected ratios for the serial dilution and spiking recovery of 9 and 6 fecal extracts ranged from 90.4 to 152.0% and from 71.3 to 112.3%, respectively. Coefficients of variation for intra- and interassay variability for 6 fecal extracts were ≤10.8% and ≤12.5%, respectively. The reference intervals for the mean and maximum Fcα1-PI from fecal samples collected on 3 consecutive days were 2.2–13.9 µg/g and 2.2–21.0 µg/g, respectively. Fcα1-PI was significantly higher in dogs <1 year of age ( P < 0.0001 for both mean and maximum Fcα1-PI for the 3 samples). The radioimmunoassay described is sensitive, linear, precise, reproducible, and accurate for clinical use, thus allowing reliable quantification of Fcα1-PI in clinical patients. Using this assay, a mean or a maximum Fcα1-PI for 3 sampling days of >13.9 µg/g or >21.0 µg/g, respectively, should be considered abnormal in dogs >1 year of age. Fecal cα1-PI concentrations in dogs <1 year of age were significantly higher and should be carefully interpreted in this age group.
Minimum inhibitory concentrations of selected antimicrobial agents for Moraxella bovoculi associated with infectious bovine keratoconjunctivitis
Authors: Angelos John A. , Ball Louise M. , Byrne Barbara A.
Published: May 2011

Infectious bovine keratoconjunctivitis (IBK) has been associated with ocular infections by Moraxella bovis, the established etiologic agent of IBK, and more recently, Moraxella bovoculi, a recently described species of Moraxella. To assist in designing rational treatment regimens for M. bovoculi infections associated with IBK, the in vitro susceptibilities of 57 M. bovoculi field isolates cultured from eyes of cattle with IBK in California from 2002 through 2007 were determined. The minimum inhibitory concentration required to inhibit the growth of 90% of organisms (MIC90) of the following 18 antibiotics tested in the present study were: danofloxacin and enrofloxacin: ≤0.12 µg/ml; ampicillin and ceftiofur: ≤0.25 µg/ml; penicillin: 0.25 µg/ml; gentamicin: ≤1 µg/ml; chlortetracycline, oxytetracycline, and tiamulin: 1 µg/ml; florfenicol: 0.5 µg/ml; trimethoprim–sulfamethoxazole: ≤2/38 µg/ml; clindamycin: 2 µg/ml; neomycin and tilmicosin: ≤4 µg/ml; tulathromycin: 4 µg/ml; spectinomycin and tylosin: 16 µg/ml; and sulfadimethoxine: >256 µg/ml. The low MIC90 of these M. bovoculi isolates suggests that commonly used antibiotics for treatment of IBK associated with M. bovis should also be effective against M. bovoculi.
Cutaneous Hepatozoon canis infection in a dog from New Jersey
Authors: Little Liz , Baneth Gad
Published: May 2011

A 7-month-old mixed-breed intact female dog was presented to a private veterinarian with a 2 cm in diameter raised, pruritic, alopecic, subcutaneous, fluctuant swelling over the right eye. Cytology of the mass revealed many degenerate neutrophils, moderate numbers of eosinophils, moderate numbers of macrophages, rare mast cells, and few erythrocytes. Rare neutrophils contained a protozoal agent compatible with a Hepatozoon gamont. Real-time polymerase chain reaction of peripheral blood was positive for Hepatozoon canis. The complete sequence identity of the amplified 18S ribosomal RNA fragment from the dog’s blood confirmed H. canis and proved it was relatively distant from the corresponding fragment sequence of Hepatozoon americanum. This case is important in documenting an unusual presentation of infection with H. canis outside of the southern United States.
An Aerogenic Pasteurella-Like Organism Isolated from Horses
Authors: Schlater Linda R. K.
Published: May 2011

Thirteen strains of a gram-negative, fermentative bacterium that produced gas from glucose were isolated from horses with a variety of clinical conditions. The morphological and biochemical characteristics of this bacterium are similar to those described for the family Pasteurellaceae. These strains appear to constitute a new taxon within the genus Pasteurella; however, the final taxonomic position of this group depends upon more detailed genetic studies. Case histories indicate that this bacterium may be a primary respiratory pathogen and may play a secondary role in various other disease conditions.
Systemic Salmonellosis in Mature Beef Cows
Authors: Morter Raymond L. , Armstrong Charles H. , Amstutz Harold E. , Thacker H. Leon
Published: May 2011

Systemic infection of mature beef cows with Salmonella typhimurium resulted in death of cows, abortions, and premature births. Salmonella typhimurium was isolated from the kidney, liver, and spleen of cows but not from an aborted fetus. Diarrhea was not a prominent clinical feature of the epizootic. The source of the salmonella was not determined.
Rapid Fluorescence Detection of in Situ Hybridization with Biotinylated Bovine Herpesvirus-1 DNA Probes
Authors: Brock Kenny V. , Potgieter Leon N. D
Published: May 2011

The use of biotinylated DNA hybridization probes for clinical detection of bovine herpesvirus-1 was investigated. Biotinylated DNA hybridization probes were prepared from bovine herpesvirus-1 DNA purified from infected cell cultures. The viral DNA was nick translated in the presence of biotin-dUTP with DNA polymerase to incorporate biotin into the newly synthesized strand. The probe was tested for specificity in in situ hybridization assays with bovine herpesvirus-1 DNA. Hybridization was detected using avidin-fluorescein single sandwich systems and an avidin-globulin with anti-globulin-fluorescein double sandwich system. Hybridization was detected by specific fluorescence of infected cells. Fluorescence was present only in bovine herpesvirus-1-infected cell culture and not in noninfected cell culture or cell cultures infected with several other viruses. The assay was performed in 6 hr.
Elevated IgG Antibody to Sarcocystis Cruzi Associated with Eosinophilic Myositis in Cattle
Authors: Ely Ray W. , Fox J. Carl
Published: May 2011

Blood sera, skeletal muscle, and cardiac muscle from 24 bovine carcasses condemned for eosinophilic myositis by US Department of Agriculture meat inspection veterinarians were compared with similar specimens from 35 random carcasses passed for human consumption. Fluorescence values determined by using a fluorometric immunoassay system were used to measure the specific antibodies to Sarcocystis cruzi. The values were significantly elevated in carcasses condemned for eosinophilic myositis as compared to carcasses passed for human consumption. The elevated fluorescence values appeared to be more than coincidental, suggesting that S. cruzi may be a causative agent of eosinophilic myositis. Microscopic examination of affected muscle revealed lesions typical of eosinophilic myositis. Lesions were characterized by extensive multifocal areas of myofiber hyaline degeneration, necrosis with sarcoplasmic fragmentation, mineralization of occasional myofibers, and atrophy of fibers with varied stages of fibrosis. Inflammatory cell exudates were predominantly eosinophils, with some macrophages and lymphocytes, and extravasated erythrocytes within, and adjacent to, affected myofibers. Affected muscles contained more S. cruzi than unaffected muscle from passed carcasses. However, a distinct cause and effect relationship could not be determined between the parasite and the presence of eosinophilic myositis.
Diagnostic Investigation of Bovine Viral Diarrhea Infection in a Minnesota Dairy Herd
Authors: Werdin Ronald E. , Ames Trevor R. , Goyal Sagar M. , DeVries Gordon P.
Published: May 2011

Bovine viral diarrhea (BVD) virus infection was diagnosed in neonatal calves with enteritis. Successful diagnostic procedures included direct immunofluorescence of frozen tissue sections, histopathology, and virus isolation. Virus isolation from buffy coats and serum was successful in detecting infected animals, whereas direct immunofluorescence of buffy coat samples was found to be less reliable. Virus was not isolated from any fecal samples. Booster vaccinations and the culling of animals shedding virus resulted in improved calf viability in this herd. It is suggested that procedures for the diagnosis of BVD virus infection should always be included in the diagnosis of neonatal calf enteritis.
Laboratory Investigation of a Naturally Occurring Outbreak of Arthrogryposis-Hydranencephaly in Texas Sheep
Authors: Crandell Robert A. , Livingston Charles W. , Shelton Maurice J.
Published: May 2011

An epizootic of abortions, weak lambs, stillbirths, and congenital arthrogryposis-hydranencephaly occurred in a sheep flock in West Texas. The outbreak began during the first week of January 1987 and continued through the third week of February 1987. Lambs born after February 1987 were not affected. A high incidence of antibodies to Cache Valley virus (Texas 7856 isolate) was demonstrated in the ewes' serum and in precolostral serum from affected lambs. No virus was isolated from tissues and body fluids of the affected lambs. The clinical, pathological, and immunological features of the epizootic were similar to those reported in Akabane virus infection in sheep. Although serological findings strongly suggest Cache Valley virus as the etiological agent of this outbreak, transmission studies are needed.
Measurement of IgG Concentration in Ovine Fetal Fluids: A Useful Diagnostic Test
Authors: Hedstrom Olaf , Sonn Robert , Dearing Peggy , Snyder Stanley P. , Lassen E. Duane
Published: May 2011

The Veterinary Diagnostic Laboratory at Oregon State University received 172 aborted ovine fetuses during the 1985–1987 lambing seasons; from 120 of these, body fluids were evaluated for IgG levels. Fifty-two (43%) of the fetal fluids had immunoglobulin G (IgG) levels greater than 15 mg/dl. Forty-five (87%) of the fluids with elevated IgG levels were confirmed or presumed toxoplasma or Chlamydia abortions. A mean fetal fluid IgG concentration of 111.5 ± 78 mg/dl was found for the 26 toxoplasma abortions; for the 19 Chlamydia abortions, a mean IgG concentration of 109 ± 91 mg/dl was found. Antibody titers equal to or greater than 1:40 against Toxoplasma gondii were detected in 23 fetal fluids. Fetal fluid IgG concentration less than 15 mg/dl was found to be associated with bacterial organisms (i.e., Campylobacter sp.) as the confirmed or presumed cause of abortion. These results suggest that measurement of fetal fluid IgG concentration is a useful, supportive diagnostic test in determining the cause of ovine abortion, and should be included as a routine laboratory procedure for ovine abortion diagnosis.
Serologic Examination of Aborted Ovine and Bovine Fetal Fluids for the Diagnosis of Border Disease, Bluetongue, Bovine Viral Diarrhea, and Leptospiral Infections
Authors: Kirkbride Clyde A. , Johnson Mark W.
Published: May 2011

Fetal serum from most of 994 bovine and 553 ovine aborted fetuses was tested serologically for antibodies to border disease (BD), bovine viral diarrhea (BVD), and bluetongue (BT) viruses, and to Leptospira sp., and the results were compared with the results of isolation procedures, fluorescent antibody tests (FAT), and histologic examinations of the same fetuses. Antibodies to BT virus were not found in any of the 994 bovine and 553 ovine fetuses. Antibody titers to BVD virus were present in 39 of 966 bovine fetuses tested, and BVD virus was detected in 4 of the 39. Four of 74 fetuses in which the BVD virus was detected by FAT or isolation had titers to BVD virus. Microagglutination (MAT) titers to 1 or more of 5 serovars of leptospires were present in 52 of 773 bovine fetal sera tested. Leptospires were not detected by FAT in any bovine fetuses that had leptospiral antibody titers. Leptospires were detected by FAT in 15 aborted calves, and none of these had MAT titers. Antibody titers to BD virus were present in 80 of 486 fetal lamb sera tested, and the virus was detected by FAT or isolation in 3 of the 80 fetuses. Border disease virus was detected in 14 of 486 fetal lambs tested. Twelve of the 14 were tested serologically and 3 had titers to BD virus. Leptospiral antibody titers were present in 27 of 326 ovine fetal sera tested. Leptospires were not detected in any of the 326 ovine fetuses tested by FAT.
Evaluation of Multivalent Leptospira Fluorescent Antibody Conjugates for General Diagnostic Use
Authors: Miller David A. , Wilson Mark A. , Kirkbride Clyde A.
Published: May 2011

Four lots of conjugate were evaluated for optimal dilution and degree of fluorescence produced with reference cultures and bovine and porcine leptospira isolates. One lot that uniformly produced better fluorescence was evaluated for sensitivity and specificity with reference cultures, isolates, culture-positive tissues, and 13 other bacterial species. Further evaluation of the conjugates was done with bovine, porcine, and ovine specimens submitted to a diagnostic laboratory. Leptospires were detected with the fluorescent antibody test (FAT) in 9 of 21 culture-positive bovine kidneys and were detected in diluted cultures when present at concentrations of 102-1O3 organisms/ml. With the exception of Treponema hyodysenteriae, FAT's of other bacterial cultures produced minimal fluorescence or were negative. Positives were characterized by moderate to brilliant fluorescence of typical cell forms, and most nonspecific fluorescence was eliminated with a flazo-orange counter-stain. The results indicated that the FAT utilizing multivalent conjugates could be used successfully as an additional method for diagnosis of leptospira infections.
Etiologic Studies on Late-Term Swine Abortions
Authors: Nielsen Judith N. , Armstrong Charles H. , Turek John J. , Nielsen Niels C.
Published: May 2011

One hundred thirty-eight swine abortions were studied in detail in an effort to identify an etiologic agent. A viral agent was implicated in 7 cases. Bacteria were isolated in less than half of the examined cases; however, in 61% of the cases, motile, filamentous organisms were observed in tissues and fluids. Although swine sera from farms experiencing reproductive problems had a high reactor rate to Leptospira bratislava antigen, electron microscopy of the observed organism revealed a wall-free prokaryote morphologically typical of the class Mollicutes.
A Bacteriologic Study of Scabby-Hip Lesions from Broiler Chickens in Texas
Authors: Scanlan Charles M. , Hargis Billy M.
Published: May 2011

Broilers from commercial flocks experiencing a 10–60% incidence of scabby-hip lesions at processing were examined, and selected skin lesions were cultured. Over 70% of the lesions were associated with traumatic excoriations, particularly on the caudal dorsal convexity of the birds. Most lesions were observed on birds that were 5 weeks of age or older. From the 27 specimens cultured, Clostridium perfringens was isolated in pure culture from 4 lesions and Staphylococcus species from 10 lesions. Pure cultures of staphylococci were recovered from 4 lesions, and 2–5 different staphylococci were isolated from 6 lesions. Eight staphylococci were identified as S. sciuri, 8 as S. simulans, 2 as S. epidermidis, 2 as S. lentus, 2 as S. warneri, 1 as S. cohnii, and 1 as S. intermedius. Fifty cutaneous specimens from 10 5-week-old normal broilers were cultured. A total of 197 isolates were identified including 65 S. sciuri, 52 S. lentus, 24 S. simulans, 12 S. hyicus, 11 S. warneri, 9 S. cohnii, 9 S. gallinarium, 8 S. xylosus, and 7 S. epidermidis.
Detection of Ehrlichia Risticii using an Avidin-Biotin-Immunoperoxidase Staining System
Authors: Breider Mike A. , Callahan Ginger , Corstvet Richard E.
Published: May 2011

An indirect immunoperoxidase procedure using a specific anti- Ehrlichia risticii monoclonal antibody and an avidin-biotin-peroxidase staining method was used to detect E. risticii antigen in infected P388D1 murine monocytes. Several different methods of cytological fixation were used, including acetone (15 min), 95% ethanol (15 min), Bouin's fixative (5 hr), and 10% buffered neutral formalin (24 hr). The E. risticii organisms were labeled effectively and identified in cells fixed with acetone and ethanol. However, infected P388D1 cells fixed in 10% formalin or Bouin's fixative required enzymatic digestion with 1.0% trypsin for 15 min at 37 C before positive results were evident. This indirect immunoperoxidase avidin-biotin staining procedure proved to be a sensitive assay for the detection of intracellular E. risticii and may be an effective diagnostic procedure for formalin-fixed paraffin-embedded tissue.
Use of in Situ Hybridization with a Biotinylated Probe for the Detection of Bovine Herpesvirus-l in Aborted Fetal Tissue
Authors: Ayers Virginia K. , Collins James K. , Blair Carol D. , Beaty Barry J.
Published: May 2011

Forty-five cases of bovine abortion were examined using in situ hybridization (ISH) with a biotinylated DNA probe specific for bovine herpesvirus-1 (BHV-1). Of the 45 cases, 16 were diagnosed as due to BHV-1, 15 were determined to be due to other causes, and 14 were of undetermined etiology. Direct comparisons between ISH and an immunoperoxidase (IP) test specific for BHV-1 were performed on formalin-fixed, paraffin-embedded tissue sections of lung, liver, kidney, spleen, thymus, and placenta; fluorescent antibody tests for BHV-1 and virus isolation were performed on fresh lung and liver. In comparison to these routine BHV-1 detection techniques, ISH had an overall sensitivity of 88.2% and a specificity of 89.3% in detecting BHV-1 in aborted fetuses. Immunoperoxidase was more sensitive than ISH with tissue sections from lung (87.5% vs. 69%), liver (92% vs. 17%), spleen, and placenta; results of the tests on tissue sections from kidney were concordant. Liver sections presented special problems in that nonspecific reactions were frequently observed with hybridization. With thymus sections, the rate of detection was higher by hybridization than by IP, but the specificity of some of these reactions could not be confirmed.
Recombinant cDNA Probe from Bluetongue Virus Genome Segment 10 for Identification of Bluetongue Virus
Authors: de Mattos Carlos Alberto , de Mattos Cecilia Cristina , Osburn Bennie Irve
Published: May 2011

Genome segment 10 of bluetongue virus (BTV) serotype 11 UC8 strain was cloned and subsequently hybridized to viral double-stranded RNA extracted from 90 field isolates of BTV serotypes 10, 11, 13, and 17; the prototype strains of BTV 2, 10, 11, 13, and 17; the prototype strain epizootic hemorrhagic disease virus (EHDV) serotype 1; and 4 field isolates of EHDV serotype 2. The 90 field isolates were obtained from different counties in California, Louisiana, and Idaho during the years 1979, 1980, and 1981. The cloned genetic probe hybridized with all the BTV samples tested, showing different degrees of cross-hybridization at the stringency conditions used in this study. This indicated that BTV genome segment 10 has conserved nucleotide sequences among the BTV serotypes 2, 10, 11, 13, and 17. No cross-hybridization signals were detected between the cloned genome segment 10 of BTV 11 UC8 strain and the prototype strain of EHDV serotype 1 and the field isolates of serotype 2. This probe recognized a wide variety of BTV isolates.
Genotypic Transitions among Bluetongue Viral Isolates from Domestic Ruminants in Colorado during 1981–1984
Authors: Collisson Ellen W. , Barber T. Lynwood , Shannon Colleen M. , Kemp Maurice C.
Published: May 2011

Two predominant electropherotypes of bluetongue virus (BTV) serotype 11 isolates from cattle during a 1981–1984 field study in eastern Colorado were characterized. The genomes of strains isolated from the first 2 years of the study had 1 predominant electropherotype (CO81), with the exception of 1 isolate that differed only in the migration of segment 3. A second electropherotype (CO83), with differences in the migration of 4 segments, coexisted in the same region during 1983 and 1984 with strains having the CO81 RNA profile. The genomes of CO81 and CO83 were also distinguishable from those of the US prototype of BTV 11. Analysis of the polypeptides of representative strains of each electropherotype by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the proteins were very similar. The occurrence of the CO81 electropherotype was apparently the result of multiple viral infections since the positions of 7 segments had faint second bands and single-banded variants were isolated after serial plaque purifications. In addition, protein 7 of 1 of the CO81 isolates and protein 7 of the single-banded variant differed as shown by reverse phase-high performance liquid chromatography of 35S-methionine-labeled tryptic peptides.
Association of Reoviridae Particles in an Enteric Syndrome of Poults Observed in Turkey Flocks during 1988
Authors: Lozano Luis-Fernando , Bickford Arthur A. , Castro Anthony E. , Swartzman-Andert Joyce , Chin Richard , Meteyer Carol , Cooper George , Reynolds Bruce , Manalac Rosa Lynn
Published: May 2011

An enteric syndrome of turkey poults, characterized by enteritis, crop mycosis, intestinal changes (pale, thin-walled ballooning with watery contents), and rickets, occurred during 1988 in 74 turkey flocks from different farms belonging to 9 California turkey growers. The flocks ranged in size from 9,000 to 120,000 birds. Pools of intestine sections from 618 birds, representing 78 field cases, were examined. Histopathological examination of the intestines showed a mild to severe atrophy with a reduced depth of crypts, which was more prominent in the distal part of the small intestine. Viral isolation attempts with primary cell cultures of chicken embryo kidney cells were negative. Examination by electron microscopy of negatively stained intestinal specimens revealed the presence of Reoviridae particles of 58.8 to 80 nm in diameter. Enzyme-linked immunosorbent assay results on the intestinal pools for mammalian and group A avian rotaviruses were negative. A statistically significant relationship was found for the presence of Reoviridae particles in the intestines of 10-21-day-old birds. Of the 7 most common pathological conditions analyzed, 2, rickets and intestinal changes (thin-walled ballooning intestine with watery contents), showed a statistically significant association with the presence of Reoviridae particles.
Leptospira Abortion in Horses
Authors: Hodgin E. Clay , Miller David A. , Lozano Fernando
Published: May 2011

Leptospira infection was diagnosed as the cause of 4 late-term equine abortions/stillbirths and 1 neonatal death in Louisiana. The most consistent gross and microscopic lesions were icterus and interstitial nephritis, respectively. Diagnoses were based on visualization of compatible spirochetes in Warthin-Starry-stained sections of kidney, liver, and placenta. Confirmation by immunofluorescence was made in 2 cases.
Diagnostic Features of Chlamydia Infection in Dairy Calves
Authors: Reggiardo Carlos , Fuhrmann Thomas J. , Meerdink Gavin L. , Bicknell Edward J.
Published: May 2011

Chlamydia were routinely isolated from a group of calves with pneumonia, systemic disease, and enteric involvement. The most commonly observed lesions were acute interstitial pneumonia, hepatitis, and enteritis. Cultures of sequentially collected nasal and rectal samples from clinically normal calves revealed a large percentage of inapparent infections during, at least, the first 30 days of age.
Blocking ELISA to Distinguish Pseudorabies Virus-Infected Pigs from Those Vaccinated with a Glycoprotein gIII Deletion Mutant
Authors: Kit Saul , Awaya Yukikazu , Otsuka Haruki , Kit Malon
Published: May 2011

A blocking enzyme-linked immunosorbent assay (ELISA) test has been developed to distinguish pseudorabies virus (PRV)-infected pigs from those immunized with a glycoprotein g92(gIII) deletion mutant, PRV(dlg92dltk). The blocking ELISA utilizes 96-well microtiter test plates coated with a cloned PRV g92(gIII) antigen, a mouse monoclonal antibody against gIII antigen (moMCAgIII): horseradish peroxidase (HRPO) conjugate, and undiluted test sera. Analyses can be completed in less than 3 hours with results printed out by an automated plate reader. Analyses on over 300 pig sera from PRV-free farms, on sera from other species, and on control sera containing antibodies to microorganisms other than PRV showed that the ratio of the optical density at 405 nm for the test sample to the optical density at 405 nm for the negative control (S/N value) was >0.7 for all sera. No false positives were identified. Likewise, the S/N values were >0.7 for over 400 sera obtained from pigs vaccinated twice with more than 1,000 times the standard PRV (dlg92dltk) dose or 1–4 times with the standard dose (2 × 105 TCID50/pig). Following challenge exposure to virulent PRV, the S/N values of the vaccinates were 0.1, showing that g92(gIII) antibodies in the sera of experimentally challenged pigs strongly blocked the binding of the moMCAgIII: HRPO conjugate to the antigen-coated wells. Sera of 233 pigs from PRV-infected herds with virus neutralization (VN) titers of 1:4 or greater were tested. All except 2 of these sera had S/N values <0.7 and more than 175 had S/N values <0.1. Sixteen sera from feral pigs with VN titers of 1:4 or greater had S/N values of 0.24 or less, but 2 sera with VN titers of 1:4 when tested 5 years prior to the PRV g92(gIII) blocking ELISA test gave false negative S/N values. Twenty-four of 29 pig sera from PRV-infected herds with VN titers < 1:4 were positive for g92(gIII) antibodies, illustrating the sensitivity of the PRV g92(gIII) blocking ELISA test. Analyses on 7 sera with VN titers of 1:4–1:64 showed that titers obtained with the PRV g92(gIII) blocking ELISA test were from 2- to 16-fold greater than the VN titers. The accuracy and sensitivity of the PRV g92(gIII) blocking ELISA test was further demonstrated by analyses of 40 unknown sera supplied in the National Veterinary Services Laboratories 1988 PRV check test kit.
Sequential Changes in the Humoral Immune Response of Pigs to Pseudorabies Virus after Vaccination, Exposure to Virulent Virus, and Reactivation of Latent Virus
Authors: Mengeling William L. , Pirtle Eugene C.
Published: May 2011

Sequential changes in the humoral immune response of pigs to pseudorabies virus (PRV) after each of several exposures to the virus were evaluated by determining virus neutralization (VN) and radioimmunoprecipitation (RIP) activities of sera collected at selected intervals. Pigs were vaccinated intramuscularly with live attenuated virus (6 pigs), inactivated attenuated virus (6 pigs), or inactivated virulent virus (6 pigs). All pigs were challenged oronasally with virulent virus 3 weeks later and 12 (4 pigs of each vaccine group) were subsequently treated with dexamethasone in an attempt to reactivate latent virus.

The relatively low serum titers of VN antibody that were raised by vaccination (titers ranged from 2 to 32) increased markedly (at least 16-fold) for all pigs after exposure to virulent virus. After dexamethasone treatment, the VN titers of 2 pigs increased 16-fold, whereas those of the other 10 dexamethasone-treated pigs and the 6 nontreated pigs either remained the same or increased only minimally (i.e., no more than 2-fold). The results of RIP using 35S-methionine-labeled viral proteins were initially similar to those of VN in that the low levels of serum RIP activity detected after vaccination increased markedly after subsequent exposure to virulent virus. In contrast to VN, however, most pigs (11 of 12) treated with dexamethasone had a clear increase in serum RIP activity. The increase was particularly striking for viral proteins of relatively low (<46K) molecular weight. Precipitating activity for 14C-glucosamine-labeled viral glycoproteins was not detected until after pigs were exposed to virulent virus. The increase in RIP activity detected after dexamethasone treatment was likely due to an additional antigenic stimulus associated with virus reactivation. However, virus was isolated from nasal swabs of only 4 of the 12 treated pigs. None of the results appeared to be affected appreciably by the type of vaccine used for initial immunization.
Isolation of Pasteurellaceae from Bovine Abortions
Authors: Ward Alton C. S.
Published: May 2011

A 2-year study was conducted to determine the incidence of Pasteurellaceae in abortion samples submitted for diagnostic evaluation. A total of 687 cases, including 623 with fetal tissues and/or stomach contents and 302 with placenta and/or uterine discharge, were evaluated. Pasteurellaceae were isolated on a nonselective medium from 9 (1.5%), 14 (2.8%), 13 (12.1%), and 42 (17.4%) of the fetal tissues, stomach contents, uterine discharges, and placentas, respectively. A total of 35 (19.9%) of 176 placental samples cultured on both a selective medium for Pasteurellaceae and a nonselective medium were positive for Pasteurellaceae. Fifteen (42.9%) of these isolates were detected only on the selective medium, whereas 5 (14.2%) were detected only on the nonselective medium and 15 (42.9%) grew on both media. Placentitis of different severity was evident in 13 (68.4%) of the 19 placentas from which Pasteurellaceae were isolated in the absence of other known abortifacient agents.
Restriction Endonuclease Analysis of Alcelaphine Herpesvirus 1 DNA and Molecular Cloning of Virus Genomic DNA for Potential Diagnostic Use
Authors: Seal Bruce S. , Klieforth Robert B. , Heuschele Werner P.
Published: May 2011

Alcelaphine herpesvirus 1 (AHV-1) genomic DNA was analyzed using restriction enzymes having recognition sequences both low in guanine-cytosine content (BamHI, KpnI, HindIII) and high in guanine-cytosine content (SmaI, AvaI, ApaI). The results from the restriction enzyme analyses along with exonuclease treatment demonstrated that the termini of AHV-1 DNA are likely composed of polyrepetitive sequences high in guanine-cytosine content similar to those found in Herpesvirus saimiri DNA. Cleaving AHV-1 DNA with the restriction enzyme Smal produced polyrepetitive sequences that appeared as low molecular weight super-molar bands < 1 kilobase pairs (kb) in size. Analysis of AHV-1 DNA cleaved with HindIII indicated that the polyrepetitive sequences are approximately 29.5 kb in size. The total molecular weight for AHV-1 DNA was determined to be approximately 118 kb. Seventy-five percent of the AHV-1 genome was cloned into the plasmids pAT153 and pBR322. Cloned DNA fragments representing unique sequences of the AHV-1 genome hybridized with AHV-1 genomic DNA but showed no appreciable hybridization with AHV-2 DNA or bovine herpesvirus 1, 2, or 4 DNA.
Identification of Brucella Abortus Strain 19 by Decreased Ability to Utilize Erythritol as Determined by Gas Liquid Chromatography
Authors: Ewalt Darla R. , Ross P. Frank , Payeur Janet B.
Published: May 2011

A method to identify Brucella abortus strain 19 by erythritol utilization using gas liquid chromatography (GLC) was developed. A total of 69 strains of B. abortus (41 virulent field strain isolates and 28 strain 19 isolates) were tested. Following incubation of the isolate with a standard amount of erythritol, the erythritol present in the cell suspension was acetylated and measured by GLC. Field strains of B. abortus utilized an average of 90.9% of the erythritol, whereas vaccine strains utilized an average of 42.4%. This difference in erythritol utilization will allow a more rapid identification of B. abortus strain 19.
A High-Performance Liquid Chromatography Method for Determination of 4-Aminopyridine in Tissues and Urine
Authors: Casteel Stan W. , Thomas Brian R.
Published: May 2011

An analytical method was developed to measure 4-aminopyridine in tissues and urine to determine appropriate diagnostic samples in acute poisoning cases. Tissues from rats dosed with 4-aminopyridine were extracted with methylene chloride. Extracts were analyzed by high-performance liquid chromatography using an isocratic solvent system of acetonitrile and aqueous solution (15/85 v/v) consisting of 0.015 M sodium salt of 1-heptane-sulfonic acid, 0.002 M tetramethylammonium bromide, and 0.01 M sodium dihydrogen phosphate adjusted to pH 3.0 with phosphoric acid. We concluded that suitable diagnostic samples for acute poisoning cases include stomach contents, kidney, liver, and urine.
Performance Studies of an Enzyme-Linked Immunosorbent Assay for Detecting Staphylococcus Aureus Antibody in Bovine Milk
Authors: Matsushita Tatsuo , Dinsmore R. Page , Eberhart Robert J. , Jones Gerald M. , McDonald John S. , Sears Philip M. , Adams D. Scott
Published: May 2011

An enzyme-linked immunosorbent assay (ELISA) for detecting Staphylococcus aureus antibody in bovine milk samples was examined for repeatability. A set of 51 bovine milk samples from 4 universities with confirmed culture results was assembled, and a panel of 30 milk samples was randomly selected. When the selected panel was tested at the collection laboratory, there was 97% agreement between the ELISA and the culture test. The panel was tested with the ELISA by the 4 university laboratories. Results were scored by both visual and optical density reader methods. When compared to reference ELISA results, the university laboratory ELISA results showed an agreement of 99.8% for negative samples, 98% for positive samples, and 99% for all samples. Additional studies on 19 milk samples that cultured positive for bacteria other than S. aureus showed 100% specificity. Overall comparison of ELISA and culture results showed high agreement between the 2 techniques. Disagreement appeared to result from explainable differences in antibody and bacterial levels and not from errors in either of the 2 techniques.
Porcine Leptospirosis in Iowa
Authors: Miller David A. , Wilson Mark A. , Owen William J. , Beran George W.
Published: May 2011

The epidemiology of leptospirosis in Iowa swine was examined on the basis of serologic results and herd data from 55 herds in the National Animal Health Monitoring System (NAHMS) program and culture results and histories from 578 cases of reproductive failure submitted to the Iowa Veterinary Diagnostic Laboratory during a 3-year period. Thirty-eight percent of sera from NAHMS herds contained antibodies against 1 or more of 12 leptospira antigens. Leptospires were isolated from 9 (1.6%) of 578 cases of reproductive failure. Seven (78%) of the isolates were identified as Leptospira interrogans serovar kennewicki and 2 (22%) as serovar grippotyphosa. In 7 herds from which leptospires were isolated, attack rates ranged from 1% to 84%. Clinical leptospirosis, characterized by reproductive failure and confirmed by isolation of leptospires, was sporadic. No significant differences in farrowing averages and reproductive problems were observed between vaccinated and nonvaccinated NAHMS herds or between herds with higher (43–63%) or lower (14–40%) percentages of animals that were serologically positive against serovar bratislava.
Comparison of Polyacrylamide Gel Electrophoresis, an Enzyme-Linked-Immunosorbent Assay, and an Agglutination Test for the Direct Identification of Bovine Rotavirus from Feces and Coelectrophoresis of Viral RNA's
Authors: Hammami Salah , Castro Anthony E. , Osbum Bennie I.
Published: May 2011

The dsRNA concentrated polyacrylamide gel electrophoresis (CPAGE) detected rotavirus directly from 19% of 77 stool specimens from diarrheic calves. A commercial enzyme-linked immunosorbent assay (ELISA) detected 25%, latex agglutination test, 23%, and polyacrylamide gel electrophoresis (PAGE), 19%. Establishing CPAGE as the “standard,” the commercial ELISA and the latex agglutination test both had higher sensitivity (84%) than PAGE (79%). However, PAGE produced the highest specificity (100%), followed by agglutination (88%) and ELISA (84%). The commercial ELISA had a slightly higher sensitivity than agglutination, PAGE, and CPAGE, but the ELISA specificity was generally lower. The latex agglutination test had a lower sensitivity than ELISA, but specificity was higher. Agglutination had similar negative predictive values (94%), compared with agglutination and PAGE, but had the lowest positive predictive value (a measure of accuracy) (70%). Agreement with CPAGE was highest for PAGE (94.8%), followed by agglutination (87%) and ELISA (84.4%). The calculated percentages of total disagreement with all other tests indicated that ELISA differed from the other rotavirus detection assays in 10.4% of the cases, agglutination in 7.8%, PAGE in 2.6%, and CPAGE in 1.3%. The 2 PAGE assays allowed the detection of atypical rotaviruses from feces based on the characteristic “super-short” migration pattern of the 11 genomic segments of rotaviruses and of other members of the Reoviridae.