bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of GN Hajna and Tetrathionate as Initial Enrichment for Salmonellae Recovery from Swine Lymph Nodes and Cecal Contents Collected at Slaughter
Authors: Harvey Roger B. , Anderson Robin C. , Farrington Leigh A. , Droleskey Robert E. , Genovese Kenneth J. , Ziprin Richard L. , Nisbet David J.
Published: May 2011

An epidemiologic survey was conducted to determine the prevalence of salmonellae in swine from 5 farms of an integrated swine operation. The purpose of this study was to evaluate the recovery efficiencies for salmonellae from swine lymph nodes and cecal contents when GN Hajna and tetrathionate were compared as initial enrichments. Salmonellae were isolated from 61% of 645 pigs at slaughter; 324 positive cultures were from lymph nodes, and 224 were from cecal contents. Frequently, pigs had salmonellae isolated from both the lymph nodes and cecal contents. Total isolations, regardless of source, were similar for GN Hajna (247) and tetrathionate (301). There was no difference ( P > 0.05) in the number of isolations from lymph nodes when GN Hajna enrichment was compared with tetrathionate enrichment (174 vs. 150). However, there was a significant ( P > 0.05) advantage of utilizing tetrathionate when compared with GN Hajna for isolations from cecal contents (151 vs. 73).
Causes of Caprine Abortion: Diagnostic Assessment of 211 Cases (1991–1998)
Authors: Moeller Robert B.
Published: May 2011

In an 8-year period, 1991–1998, 217 accessions of caprine abortions were submitted to the California Veterinary Diagnostic Laboratory System. Of these 217 submissions, 211 were suitable for examination in this study (6 had insufficient data). Infectious agents as the cause of abortions were found in 37% of the cases: bacterial agents were identified in 30.5%, viral agents in 2%, fungal agents in 0.5%, and protozoal agents in 4% of the cases submitted. The most common causes of abortions were Chlamydia psittaci and Coxiella burnetii infection, which accounted for 23% of all goat abortions. Mineral deficiencies were observed in 4%, fetal anomalies accounted for 3%, and leukoencephalomalacia of the brain (probable oxygen deprivation) accounted for 3% of the submissions. No diagnosis was made in 112 of the 211 submissions (53%). No lesions were noted in 104 of the submissions (49%). The other 8 submissions (4%) had histologic lesions suggestive of a bacterial agent; however, no infectious agents were identified in these cases.
Comparison of Porcine Reproductive and Respiratory Syndrome Virus Growth in Media Supplemented with Fetal Bovine Serum or a Serum Replacement
Authors: Schommer Susan K. , Carpenter Susan L. , Paul Prem S.
Published: May 2011

Commercially available serum replacements are often used in cell culture as a cheaper and less variable substitute for fetal bovine serum (FBS). The growth of porcine reproductive and respiratory syndrome virus (PRRSV) isolates in CRL11171 cells maintained in a medium supplemented with FBS was compared with virus propagation in the same cell line maintained in the same medium with a serum replacement. The PRRSV replicated significantly better when the cell culture medium was supplemented with FBS. The results of this study have implications for the use of serum replacement-supplemented medium for PRRSV diagnosis by virus isolation.
Characterization of an Undifferentiated Malignancy as a Mast Cell Tumor Using Mutation Analysis in the Proto-Oncogene c-KIT
Authors: Zemke Daniel , Yamini Behzad , Yuzbasiyan-Gurkan Vilma
Published: May 2011

A 6.5-year-old female Boxer was euthanized and presented for necropsy following rapid clinical decline concomitant with the development of numerous tumor masses. The largest of these masses was in the same location as a mast cell tumor that had been previously removed from this dog. Gross examination revealed the presence of nodules 5–200 mm in diameter throughout the body, including the lymph nodes. Histologic analysis showed an influx of round cells with no granules, leading to the provisional diagnosis of systemic lymphosarcoma. Immunohistochemical staining for B- and T-lymphocyte antigens was negative. Molecular tests were used to identify a tandem duplication in the c- KIT proto-oncogene from both the earlier mast cell tumor and the current nodules, implicating a common origin. Addition of molecular testing to conventional necropsy evaluations allowed a definitive diagnosis of mast cell tumors.
Prevalence of EaeA+ Escherichia Coli Isolated from Pigs with Diarrhea
Authors: Kim Junghyun , Choi Changsun , Chae Chanhee
Published: May 2011

A total of 600 Escherichia coli strains isolated from 382 preweaned and 197 postweaned pigs with diarrhea were tested for the presence of the eaeA gene by polymerase chain reaction techniques. Of the 393 isolates from preweaned pigs, 23 (5.8%) E. coli strains isolated from 23 pigs carried the eaeA gene. Of the 207 isolates from postweaned pigs, 9 (4.3%) E. coli strains isolated from 9 pigs carried the eaeA gene. The results suggest that eaeA+ E. coli is associated with diarrhea in pre- and postweaned pigs.
Application of Competitive Enzyme-Linked Immunosorbent Assay for the Serologic Diagnosis of Classical Swine Fever Virus Infection
Authors: Clavijo Alfonso , Lin Min , Riva José , Zhou En-Min
Published: May 2011

A competitive enzyme-linked immunosorbent assay (C-ELISA), based on a truncated E2 recombinant protein of the Alfort/187 strain of classical swine fever virus (CSFV) and a specific monoclonal antibody M1669, was evaluated using 2,000 sera from clinically healthy pigs in Canada (a CSFV-free country) and sera from experimentally infected pigs. The relative specificity and sensitivity of the C-ELISA were 100% and 86%, respectively, at a cutoff of 25% inhibition using negative and positive pig sera, as defined by the neutralizing peroxidase-linked assay (NPLA). A kappa value of 0.91 was obtained, indicating an excellent level of agreement between the NPLA and the C-ELISA. When sera from 120 infected pigs were used in the test at ≥21 days postinfection, the sensitivity of the C-ELISA and the kappa value increased to 97% and 0.98, respectively. This C-ELISA will be useful when a large number of samples must be tested, as could occur during a disease outbreak or for surveillance or prevalence studies.
The Use of Polymerase Chain Reaction Assay to Diagnose Proliferative Gill Disease in Channel Catfish (Ictalurus Punctatus)
Authors: Whitaker Julia W. , Pote Linda M. , Khoo Lester , Shivaji Renuka , Hanson Larry A.
Published: May 2011

To assess the potential of a polymerase chain reaction (PCR) assay as a diagnostic tool in the detection of proliferative gill disease (PGD) in channel catfish ( Ictalurus punctatus), PCR assays were compared with the traditional diagnostic methods of gill wet mounts and histology. A PCR assay using primers for Aurantiactinomyxon ictaluri, the actinospore associated with PGD, was performed with tissues from fish from commercial ponds. Using histology as the “gold standard,” the sensitivity, specificity, and accuracy of the PCR assay were all >90%. In comparison, the wet mount examinations had a lower sensitivity and specificity. Using the chi-square test and a test for strength of association, there was a significant, strong association between results obtained by PCR and those obtained by the other 2 methods. These results demonstrate that the PCR assay is a good diagnostic tool for the detection of PGD.
Longitudinal Evaluation of Bovine Mammary Gland Health Status by Somatic Cell Counting, Flow Cytometry, and Cytology
Authors: Rivas Ariel L. , Quimby Fred W. , Blue Julia , Coksaygan Ozden
Published: May 2011

Bovine mastitis phases induced by Staphylococcus aureus were assessed in 6 lactating cows before challenge and at 1, 4–8, and 9–14 days postinoculation (dpi). Milk lymphocytes, macrophages, and polymorphonuclear cells (PMN) were counted by conventional (manual) cytology, identified by CD3+ and CD11b+ immunofluorescence and counted by flow cytometry (based on leukocyte forward and side light scatter values). Somatic cell counts (SCC) and recovery of bacteria were recorded at the same times. Preinoculation samples showed a lymphocyte-dominated composition. At 1 dpi, the percentage of PMN increased and that of lymphocytes decreased. At 4–8 dpi, PMN were predominant, but the percentage of mononuclear cells increased above that at 1 dpi and further increased by 9–14 dpi (when lymphocytes approached prechallenge values). Based on leukocyte percentages, 3 indices were created from the data: 1) the PMN/lymphocyte percentage ratio (PMN/L), 2) the PMN/macrophage percentage ratio (PMN/M), and 3) the phagocyte (PMN and macrophage)/lymphocyte percentage ratio (Phago/L). Significant correlations were found between cytologic and flow cytometric data in all of these indicators (all with P ≤ 0.01). These indices identified nonmastitic, early inflammatory (1–8 dpi), and late inflammatory (9–14 dpi) animals. In contrast, SCC and bacteriology did not. Although sensitivity of the SCC was similar to that of Phago/L, the specificity of SCC was almost half that of the Phago/L index. Based on flow cytometry indicators, an algorithm for presumptive diagnosis of bovine mastitis was developed. Flow cytometry provides results as valid as those obtained by conventional (manual) cytology, shows greater ability to identify mastitic cases than does SCC, and may identify 3 mammary gland health-related conditions.
Soft Tissue Sarcomas in the African Hedgehog (Atelerix Albiventris): Microscopic and Immunohistologic Study of Three Cases
Authors: Ramos-Vara Josá A.
Published: May 2011

Three soft tissue tumors from 2 female hedgehogs were examined microscopically and immunohistochemically. Two tumors involved haired skin and the third one was vaginal. Microscopically, the cutaneous tumors had features of malignant peripheral nerve sheath tumor (MPNST), whereas the vaginal tumor was classified only as a spindle cell sarcoma. Immunohistochemically, all 3 tumors were strongly positive for vimentin and strongly to moderately positive for CD10 and neuron-specific enolase but did not stain with antibody to S100 protein, an antigen typically present in human MPNST. The cutaneous tumor from hedgehog no. 1 was examined ultrastructurally and the neoplastic cells resembled fibroblasts. Hedgehog no. 1 was euthanized at the time of the biopsy. The outcome of the other hedgehog was unknown.
The Ovine Respiratory Syncytial Virus F Gene Sequence and its Diagnostic Application
Authors: Eleraky Nasser Z. , Kania Stephen A. , Potgieter Leon N. D.
Published: May 2011

Ruminant respiratory syncytial viruses (RSVs) are classified into 2 subgroups, ovine RSV and bovine RSV. Although ovine RSV infects cattle, its contribution to bovine respiratory tract disease has not been established, which is an important issue for vaccine development in cattle. Diagnosis by virus isolation or serology has low or variable sensitivity and/or specificity and polymerase chain reaction (PCR) has been recommended as a rapid and sensitive technique for RSV detection. A simple procedure has been developed to detect and identify bovine and ovine RSVs. First, the nucleotide sequence of the ovine RSV fusion (F) gene was determined and compared with representative strains of bovine RSV and human RSV subgroups A and B. The ovine RSV F gene has 85 and 72–73% nucleotide identity with those of bovine RSV and human RSV, respectively. The predicted amino acid sequence of the ovine RSV F gene has 94 and 83–84% amino acid identity with those of bovine RSV and human RSV, respectively. Then PCR primers targeting a specific F gene fragment of bovine and ovine RSV were designed. The primers represented bases 85–103 and the complementary sequence to bases 510–493 of the ovine RSV F gene. A similar PCR product (426 bp) was obtained on agarose gel electrophoresis from bovine RSV and from ovine RSV. The products, however, were unique to the parent virus and could be distinguished by EcoRI or MspI restriction endonuclease cleavage. EcoRI cleaved the ovine product into 2 bands (285 and 141 bp) but failed to affect the bovine RSV PCR product. However, MspI cleaved the bovine product into 2 bands (229 and 197 bp) but had no effect on the ovine product. Also, this assay did not amplify any PCR product with human RSV. The reverse transcription-polymerase chain reaction (RT-PCR) followed by restriction enzyme digestion is a useful and practical approach for detection and differentiation of ruminant respiratory syncytial viruses.
Development of a PCR Test to Diagnose Haemophilus Parasuis Infections
Authors: Oliveira Simone , Galina Lucina , Pijoan Carlos
Published: May 2011

A polymerase chain reaction (PCR) test was developed in order to improve the accuracy and speed of diagnosis of Haemophilus parasuis, an economically important respiratory pathogen that affects swine. The gene sequence of the 16S small subunit ribosomal RNA of H. parasuis (GenBank M75065) was compared with 56 16S sequences of related bacteria, including those frequently isolated from pig tissues. Two species-specific primers were designed: HPS forward and HPS reverse. The predicted size of the amplified PCR product was 821 bp. The PCR test could detect a minimum of 102 bacteria and 0.69 pg of DNA. Thirty-one H. parasuis isolates, including 12 different serovars and 19 field isolates, were positive using the PCR test. No amplification was observed when the test was run using DNA from 15 other bacterial species commonly isolated from swine tissues. A weak band was observed when the PCR test was performed using Actinobacillus indolicus DNA as template. Clinical samples tested by PCR included tissues and swabs from 5 animals naturally infected with H. parasuis and 1 experimentally infected animal. The PCR was positive in 26 of 30 clinical samples. Four samples showed weak bands, and these results were not considered positive. Haemophilus parasuis was isolated from 18 of 30 of these samples. Tissues from specific pathogen-free (SPF) pigs and from unrelated species were negative for H. parasuis isolation and PCR. The developed PCR was successfully used in the diagnosis of H. parasuis infection, especially when compared with traditional microbiology techniques.
Cultivation of Ehrlichia Canis in a Continuous BALB/C Mouse Macrophage Cell Culture Line
Authors: Keysary Avi , Waner Trevor , Strenger Carmella , Harrus Shimon
Published: May 2011

This report describes the successful adaptation of the Israeli isolate of Ehrlichia canis on a continuous mouse macrophage cell line (J774.A1). Successful infection of the J774.A1 cells was first judged by the direct immunofluorescence antibody test using an anti- E. canis-IgG:FITC conjugate. A particular property of infected J774.A1 cells was the ability to reestablish after harvesting of the monolayer by scaping. Infected cells were used as antigen for immunofluorescence antibody tests (IFA), and the results compared well with those of DH82 cells. It was concluded that the J774.A1 continuous cell line could serve as an alternate propagation cell line for E. canis organisms.
Tuberculin Skin Testing in White-Tailed Deer (Odocoileus Virginianus)
Authors: Palmer Mitchell V. , Whipple Diana L. , Waters W. Ray
Published: May 2011

The comparative cervical skin test for antemortem diagnosis of tuberculosis was done 169 times on 116 different white-tailed deer of known Mycobacterium bovis infection status. The sensitivity and specificity were 97 and 81%, respectively. The magnitude of change in skin thickness at test sites was not significantly influenced by dosage of inoculum, dissemination of the disease process, or repeated skin testing. However, the magnitude of change in skin thickness was significantly greater in deer infected for less than 109 days than in deer infected for more than 109 days. As used in the present study, the comparative cervical skin test is a sensitive method of antemortem diagnosis of M. bovis infection in white-tailed deer.
The 14-3-3 Cerebrospinal Fluid Immunoassay Lacks Utility in the Diagnosis of Clinical Scrapie
Authors: Tyler Jeff W. , Lakritz Jeff , Weaver Dusty , Johnson Gayle , VanMetre David , Smith K. , Taylor William , Jenny A.
Published: May 2011

This study determined whether the immunoassay for cerebrospinal fluid 14-3-3 protein concentration was sensitive and specific in the diagnosis of naturally occurring clinical scrapie in sheep. Cerebrospinal fluid was collected from 9 sheep with the confirmed diagnosis of scrapie. Additionally, cerebrospinal fluid was collected from 13 clinically normal sheep, which originated from a closely monitored flock with no history of scrapie. Sensitivity and specificity were calculated using standard epidemiological methods. Cerebrospinal fluid immunoassay results did not differ significantly between positive and negative sheep. Test sensitivity varied from 0.55 to 0.66, depending on the choice of test endpoint. Test specificity varied from 0.30 to 0.77, depending on the choice of test endpoint. The 14-3-3 cerebrospinal fluid immunoassay appears to have no value in the diagnosis of clinical scrapie in sheep.
A Comparison of Virus Isolation, Immunohistochemistry, Fetal Serology, and Reverse-Transcription Polymerase Chain Reaction Assay for the Identification of Porcine Reproductive and Respiratory Syndrome Virus Transplacental Infection in the Fetus
Authors: Benson James E. , Yaeger Michael J. , Christopher-Hennings Jane , Lager Kelly , Yoon Kyoung-Jin
Published: May 2011

Virus isolation (VI), immunohistochemistry (IHC), fetal serology, and reverse-transcription polymerase chain reaction assay (RT-PCR) were performed on samples from 107 fetuses comprising 10 litters taken from sows experimentally infected with porcine reproductive and respiratory syndrome virus (PRRSV). In addition to comparing the relative sensitivity and specificity of each test, RT-PCR was evaluated with respect to the relative suitability of thoracic fluids and tissues as samples, the effects of autolysis, and the effects of pooling of fetal specimens. VI, IHC, and fetal serology identified PRRSV infection in 48.6%, 23.4%, and 14.9% of 107 fetuses, respectively, and identified at least 1 infected fetus in 10, 10, and 5 of 10 litters, respectively. In utero death with autolysis reduced the test efficacy of all 3 methods. Fetal thoracic fluid and tissues were equally suitable for RT-PCR detection of PRRSV Pooling fetal tissues or fluids from VI-positive animals with comparable material from negative controls had no detrimental effect on RT-PCR results when evaluated at dilutions of 1:1, 1:2, 1:4, and 1:8. The results of RT-PCR testing were positive in 100%, 94.4%, and 83.3% of VI-positive specimens allowed to autolyze at 4, 21, or 37 C, respectively, for 24, 48, and 96 hours. Compared with the other testing modalities, RT-PCR appeared to be impacted the least by the adverse effects of autolysis.
Prognostic Value of Histologic Stage and Proliferative Activity in Canine Malignant Mammary Tumors
Authors: Sarli Giuseppe , Preziosi Rosario , Benazzi Cinzia , Castellani Gastone , Marcato Paolo Stefano
Published: May 2011

The aim of this study was to investigate the correlation between the histologic invasiveness (histologic stage) and various cell proliferation activity assays (quantity of argyrophil proteins associated with nucleolar organizer regions [AgNORs], mitotic activity, MIB1 [Ki67] immunohistochemical detection) for predicting the biologic behavior of malignant canine mammary tumors. Sixty specimens from malignant canine mammary tumors with no distant metastases (M0) at surgery were selected, and follow-up data were collected over a 2-year period. The histologic invasiveness was graded by histologic stage (stage 0 = tumors without stromal invasion; stage I = tumors with stromal invasion; stage II = tumors with neoplastic emboli in vessels), and the proliferative indices were expressed as MIB1 index (the percentage of nuclear area immunohistochemically stained by MIB1 antibody), mitotic index (the number of mitoses per 1,000 neoplastic cells), and AgNOR index (the ratio between mean AgNOR area of tumor cells and the mean AgNOR area of fibroblasts/lymphocytes). The measures of proliferative activity were compared among groups with different histologic stages, and the influence of different prognostic variables (histologic stage, AgNOR index, mitotic index, MIB1 index) on survival time was evaluated. A significant difference in the proliferation patterns was recorded between the different histologic stages for the mitotic index ( P = 0.0006) and MIB1 index (0.0013). Among the different parameters considered, histologic stage ( P < 0.05), AgNOR index ( P = 0.0291), and MIB1 index ( P = 0.014) revealed a significant association with prognosis in univariate analysis. AgNOR index for 1-year survival and histologic stage for 2-year survival were the most significant parameters influencing survival, as determined by multiple nonlinear logistic regression.
Scirrhous Gastric Carcinoma with Mediastinal Invasion in a Dog
Authors: Wang Fun-In , Lee Jihjong J. , Liu Chen-Hsuan , Pang Victor Fei , Chang Pen-Heng , Jeng Chian-Ren
Published: May 2011

An 8-year-old male Rottweiler was presented for recurrent episodes of dysphagia and vomiting with chronic weight loss. Radiography revealed a mediastinal mass in the heart base region. Necropsy revealed a firm, white mediastinal mass extending along the distal esophagus, through the diaphragm, to the gastric cardia, leftward to the convex visceral aspect of the fundus, and rightward along the lesser curvature of the stomach to the pyloric antrum. The gastric lymph node was enlarged and the omentum contained several nodules. Histologically, deep fundic mucosa contained pleomorphic, vacuolated cells with intracytoplasmic mucin, which was hyaluronidase resistant. Neoplastic cells were cytokeratin positive and vimentin negative. Transmural invasion was evidenced by the presence of cytokeratin-positive cells between smooth muscle bundles of the gastric wall. The mediastinal mass was composed of clusters of neoplastic cells in a stroma of dense and loose connective tissue. Neoplastic cells were also within blood and lymphatic vessels, tracheobronchial and gastric lymph nodes, and around peripheral nerves. This carcinoma most likely arose from the gastric fundus and extended to the cardia, from where it advanced proximally to the mediastinum as well as further rightward along the lesser curvature, demonstrating an anatomic continuity suggestive of a direct invasion. Metastasis, evidenced by the presence of lymphatic, blood, and perineural tumor emboli, also occurred.
Confirmation of Vertical Transmission of Bovine Immunodeficiency Virus in Naturally Infected Dairy Cattle Using the Polymerase Chain Reaction
Authors: Moody Cary A. , Pharr G. Todd , Murphey Joey , Hughlett Marian B. , Weaver Carroll C. , Nelson Phillip D. , Coats Karen S.
Published: May 2011

The purpose of this study was to determine whether bovine immunodeficiency virus (BIV) is vertically transmitted in naturally infected dairy cattle. Twenty-two dam/calf pairs from a Mississippi Agriculture and Forestry Experiment Station dairy were the study group. Blood samples were collected following delivery of calves, the peripheral blood leukocytes were purified from these samples, and the leukocyte DNA was used in polymerase chain reactions targeting the pol gene region of the BIV provirus. Southern blotting and hybridization were used to confirm the BIV specificity of the amplified fragments. BIV provirus was detected in 14 of 22 calves (64%), demonstrating vertical transmission. Eight of the calves were disqualified from the final interpretation of transplacental transfer because they may have nursed their mothers prior to blood collection, allowing the possibility of lactogenic transfer of the virus. Transplacental transmission of BIV was identified in 6 of 22 calves (27%).
Detection of Bovine Viral Diarrhea Virus by TaqMan® Reverse Transcription Polymerase Chain Reaction
Authors: Mahlum Carrie E. , Haugerud Sigrun , Shivers Jan L. , Rossow Kurt D. , Goyal Sagar M. , Collins James E. , Faaberg Kay S.
Published: May 2011

Detection and elimination of calves and cows persistently infected with bovine viral diarrhea virus (BVDV) is important for the control of this pathogen. Historically, BVDV detection involved cell culture isolation followed by virus detection through immunofluorescence or immunoperoxidase monolayer assay (IPMA) methods. More recently, immunohistochemistry (IHC) has been added as a routine test for BVDV detection. The detection of BVDV by gel-based reverse transcription polymerase chain reaction (RT-PCR) is more sensitive and rapid than by cell culture isolation, but test results can be compromised by sample contamination during nucleic acid amplification. This study was designed to develop a closed-tube format of BVDV nucleic acid amplification and detection, TaqMan® RT-PCR. The results of this new technique were compared with those obtained with virus isolation, IPMA, and IHC. With TaqMan® RT-PCR, BVDV was detected in many samples negative by IPMA, IHC, and virus isolation with the exception of 1 sample that was positive by IHC. TaqMan® RT-PCR in a closed-tube format offers a rapid, economical, high volume, and sensitive method for BVDV detection without the concerns of amplified cDNA product contamination associated with open-tube gel-based PCR tests.
Effects of Different Variables on whole Blood Cholinesterase Analysis in Dogs
Authors: Tecles Fernando , Panizo Cándido Gutiérrez , Subiela Silvia Martínez , Cerón Jose J.
Published: May 2011

The influence of several variables such as sample and reagent storage, anticoagulants, reaction temperature, pH, and substrate concentration on whole blood cholinesterase determination was studied. Storage of nondiluted whole blood samples at room temperature or under refrigeration (4 C) was adequate for short-term storage (3 days to 2 weeks). However, freezing would be more appropriate for long-term storage (≥1 month), and successive thawing and freezing did not produce any loss of cholinesterase activity. All reagents (2,2′-dithiodipyridine as chromophore and acetylthiocholine and butyrylthiocholine as substrates) were stable for 3 months when frozen. Heparin and ethylenediaminetetraacetic acid were the most suitable anticoagulants for whole blood acetylcholinesterase and butyrylcholinesterase determination, because citrate yielded lower acetylcholinesterase values and fluoride inhibited butyrylcholinesterase. Increases in reaction temperature and pH yielded higher cholinesterase values but also increased nonenzymatic substrate hydrolysis. Higher cholinesterase and nonenzymatic substrate hydrolysis values were obtained as higher substrate concentrations were used.
A Familial Degenerative Neuromuscular Disease of Gelbvieh Cattle
Authors: Moisan Peter G. , Steffen David J. , Sanderson Michael W. , Nietfeld Jerome C. , Finley Melissa R. , Grotelueschen Dale M. , Andrews Gordon A. , Johnson Gayle , Williamson Lisa , Rushton Steven D. , Hall D. Greg , Harmon Barry G.
Published: May 2011

A degenerative skeletal muscle disease with vascular, neurologic, and renal lesions and a probable familial distribution was identified in 4–20-month-old purebred Gelbvieh cattle. Thirteen affected animals were confirmed from 6 separate beef herds, with a mortality rate of 100%. Clinical signs in affected animals consisted of ataxia, weakness, and terminal recumbency. Gross and histologic muscle lesions were indicative of nutritional myopathy of ruminants, with a lack of myocardial lesions in most cases and only rare myocardial changes in a few animals. Acute to chronic lesions in most large skeletal muscle groups consisted of degeneration, necrosis, regeneration, fibrosis, and atrophy. Fibrinoid necrosis of arterioles was a common feature in multiple tissues. Lesions in the spinal cord white matter and peripheral nerves consisted of degeneration of the dorsal columns and axons, respectively. Changes in the kidneys consisted of chronic interstitial nephritis with fibrosis, hyaline droplet change and tubular epithelial vacuolar change and were most severe in the older calves. Intracytoplasmic myoglobin and iron were demonstrated within the hyaline droplets in degenerate renal cortical tubular epithelial cells. Vitamin E levels were deficient in most (6/7) of the animals tested. Investigation of the pedigree of affected animals revealed a common ancestry for all but 1 of the animals whose parentage could be traced. This investigation suggests that a hereditary metabolic defect, possibly involving antioxidant metabolism, could be responsible for this condition. Renal disease, possibly secondary to myoglobinuria, may be unique to this bovine condition.
Placentitis, Fetal Pneumonia, and Abortion Due to Rhodococcus Equi Infection in a Thoroughbred
Authors: Patterson-Kane Janet C. , Donahue J. Mike , Harrison Lenn R.
Published: May 2011

Rhodococcus equi is a rare cause of equine abortion. This report describes pyogranulomatous placentitis and fetal pneumonia in a case of abortion from a Thoroughbred mare. Numerous Gram-positive coccobacilli were noted histologically within macrophages in placental and pulmonary lesions. Rhodococcus equi was isolated in pure culture from the placenta, lung, liver, kidney, and stomach content. This is the first description of placentitis due to Rhodococcus equi infection in a horse.
Immunohistochemical Demonstration of Francisella Tularensis in Lesions of Cats with Tularemia
Authors: DeBey Brad M. , Andrews Gordon A. , Chard-Bergstrom Cindy , Cox Linda
Published: May 2011

An immunohistochemical test was developed and validated for detection of Francisella tularensis antigen in tissues of cats with fatal tularemia. Ten cases of naturally occurring tularemia in cats were positive both by isolation of F. tularensis and immunohistochemical identification of F. tularensis antigen. Nine additional cases with lesions typical of tularemia were positive for F. tularensis antigen, although bacterial cultures were not performed. Immunohistochemical identification of F. tularensis in formalin-fixed tissue is valuable for establishing a rapid etiologic diagnosis under circumstances where fresh tissues may not be available for isolation and identification of the organism.
Diagnosis of Epizootic Bovine Abortion in Nevada and Identification of the Vector
Authors: Hall Mark R. , Hanks Donald , Kvasnicka William , Bosomworth Alan , Smith Harry , Stott Jeffrey L. , Blanchard Myra T. , Anderson Mark L.
Published: May 2011

In the 43 years since the first description in California, epizootic bovine abortion (EBA) has been considered but not definitively diagnosed as a cause of late-term abortions on Nevada ranches. Examination of aborted full-term bovine fetuses obtained from Nevada ranches revealed gross abnormalities consistent with EBA (enlarged lymph nodes, petechial hemorrhages of the oral mucosa and conjunctiva, ascites, and splenohepatomegaly), and EBA was confirmed by histologic examination of fetal tissues. The histologic thymic changes were characteristic of EBA and included severe histocytic thymusitis with depletion of thymocytes, interlobular hemorrhage, and fibrinocellular exudation. The gross enlargement of lymph nodes was the result of cortical follicular hyperplasia and histiocytic lymphadenitis. In addition, widespread, predominately nonsuppurative histologic lesions typical of EBA were observed in most organs, including the brain, lung, heart, liver, and spleen. Furthermore, the presence of Ornithodorus coriaceus, the argasid tick vector of EBA, was established by tick collection using CO2 traps. The tick was identified on ranches and in geographic areas (northern and northwestern counties of Nevada) coincident with diagnosis of multiple cases of EBA. This study establishes the presence of EBA as a cause of late-term abortion in Nevada. Additionally, identification of the EBA tick vector, O. coriaceus, in the same areas as the abortions provides strong evidence that the disease is endemic.
Evaluation of Kinetics and Single-Read Enzyme-Linked Immunoassays for Detection of Toxoplasma Gondii Antibodies in Sheep
Authors: Werre Stephen R. , Jacobson Richard H. , Bowman Dwight D. , Dubey Jitender P. , Mohammed Hussni O.
Published: May 2011

A kinetics enzyme-linked immunosorbent assay (ELISA) and a single-read ELISA for the detection of ovine anti- Toxoplasma gondii IgG were developed and optimized. During the kinetics assay, 3 optical densities were obtained for each serum sample at intervals of 45 seconds, and the results were presented as average slopes (replicates of 2) of the reaction rate between bound enzyme and substrate solution. The kinetics ELISA was stopped 5 minutes after dispensing the substrate to constitute the single-read ELISA, and the results were presented as average optical densities for duplicates of each sample. Performance of the assays was evaluated using the modified agglutination test (MAT) as the “gold standard.” There was a high level of agreement between both ELISAs and the MAT, as measured by Pearson correlation coefficients, kappa statistics, and the area under the receiver operating characteristics curves. The single-read ELISA was as accurate as the kinetics ELISA, with a sensitivity of 89% and a specificity of 96%.
Detection of Infectious Bursal Disease Virus in Different Lymphoid Organs by Single-Step Reverse Transcription Polymerase Chain Reaction and Microplate Hybridization Assay
Authors: Barlič-Maganja Darja , Zorman-Rojs Olga , Grom Jože
Published: May 2011

A rapid and sensitive method for the detection of infectious bursal disease virus (IBDV) RNA in different chicken lymphoid organs was developed. The method is based on a single-step reverse transcription polymerase chain reaction (RT-PCR) procedure and the enzyme-linked immunosorbent assay (ELISA) detection method of amplified products. Vaccinal IBDV strain and field isolates were used for the optimization of RT-PCR and for the determination of conditions for microplate hybridization and colorimetric detection of the amplicons. With this method, viral RNA could be detected in various stages of infection in samples of different lymphoid organs. Bursas and cecal tonsils were suitable organs for viral RNA detection at different times during IBDV infection. The RT-PCR/ELISA method can be applied for IBDV detection in routine diagnostic tests, which are not usually carried out because of the difficulties involved in isolating the virus.
Evaluation of Nested Polymerase Chain Reaction, Pepsin/Trypsin Digest, and Histology for the Detection of Myxobolus Cerebralis in Salmonid Fishes of Indiana and Michigan
Authors: Qureshi T. , White M. R. , Santrich C.
Published: May 2011

The objectives of this study were to survey fish from state hatcheries in Indiana and Michigan and to compare the nested polymerase chain reaction (PCR) test with pepsin/trypsin digest (PTD) and histopathology for the diagnosis of whirling disease (WD). One group of 40 and 9 groups of 60 fish heads, for a total of 580 samples, were submitted from hatcheries in Indiana and Michigan. These samples were examined for myxozoan spores using histopathology, PTD, and PCR tests. The heads were hemisectioned, and one half was fixed in 10% neutral-buffered formalin for histopathologic examination. The other half was processed for PTD. Some of the sediment was examined for the presence of myxozoan spores, and the rest was prepared for the nested PCR. Histologic examinations did not reveal Myxobolus cerebralis in any of the 580 samples. One hundred serial step sections, taken at 5-μm intervals, were evaluated for samples with positive spore identification by PTD. Histologic examination of these sections failed to reveal any myxozoan parasites. Myxozoan spores were observed in 16.9% (98/580) of samples in sediment after PTD. Spores morphologically similar to those of M. cerebralis were observed in 1.0% of PTD samples ( n = 6). The nested PCR indicated that M. cerebralis spores were present in 0.5% of samples ( n = 3). All 3 nested PCR-positive samples came from the same hatchery; however, spores of M. cerebralis were seen in 1 sample, spores of other myxozoan species were seen in the second sample, and spores were not seen in the third sample. When comparing the PTD to the nested PCR test, the PTD diagnosed 1 true positive, 5 false positives, 2 false negatives, and 572 true negatives, for a sensitivity of 33% and a specificity of 99.1%. Screening for M. cerebralis infection in this study indicated a low prevalence of the disease. Histopathology was a very insensitive indicator of WD. The PCR test was highly specific and was used to differentiate spores of M. cerebralis from similar spores of other species.
Association of Two Newly Recognized Herpesviruses with Interstitial Pneumonia in Donkeys (Equus Asinus)
Authors: Kleiboeker Steven B. , Schommer Susan K. , Johnson Philip J. , Ehlers Bernard , Turnquist Susan E. , Boucher Magalie , Kreeger John M.
Published: May 2011

Over a period of 6 years, antemortem and postmortem examinations were performed on a number of donkeys suffering from respiratory disease. For many cases, initial diagnostic efforts failed to identify an etiology consistent with the pathologic findings. However, retrospective examination of these cases using consensus primer polymerase chain reaction, designed to recognize herpesviruses from all 3 subfamilies of the Herpesviridae, amplified a fragment of the highly conserved herpesvirus DNA polymerase gene from a number of these animals. Two novel herpesviruses, herein designated asinine herpesvirus 4 (AHV4) and asinine herpesvirus 5 (AHV5), were consistently detected in lung tissue from donkeys in which the histopathology was characterized by interstitial pneumonia and marked syncytial cell formation but not in lung tissue from donkeys with evidence of bacterial or verminous pneumonia. Nucleotide sequence and phylogenetic analysis places these new viruses within the Gammaherpesvirinae subfamily and indicates that they are most closely related to the recently identified zebra herpesvirus and wildass herpesvirus as well as equine herpesviruses 2 and 5.
Evaluation of Microbial Culture Techniques for the Isolation of Pythium Insidiosum from Equine Tissues
Authors: Grooters Amy M. , Whittington Amy , Lopez Mae K. , Boroughs Michelle N. , Roy Alma F.
Published: May 2011

The purpose of this study was to evaluate the effects of sample handling, storage, and culture techniques on the isolation of Pythium insidiosum from infected equine tissues. Tissue and kunker samples obtained immediately posteuthanasia from a horse with subcutaneous pythiosis were used to assess the effects of sample type (kunkers vs. tissues), media type (selective vs. nonselective), storage technique, and storage time on P. insidiosum isolation rate. Overall, isolation rates were higher from fresh kunkers (94.6%) and stored kunkers (76.4%) than from fresh tissues (8.3%) or stored tissues (4.6%). Isolation of P. insidiosum also occurred more often on antibiotic-containing media than on nonselective media for both fresh and stored samples. For samples that were stored for 1–3 days prior to culture, P. insidiosum isolation rates were highest for the following techniques: kunkers stored at room temperature and plated on selective media (100%), kunkers stored at 4 C and then plated on either nonselective (91.7%) or selective (95.8%) media, kunkers stored on cold packs and then plated on either nonselective (93.8%) or selective (100%) media, kunkers stored in ampicillin solution and plated on selective media (100%), and kunkers stored in ampicillin/gentocin solution and plated on selective media (87.5%). For samples stored for 4–5 days, P. insidiosum isolation rates were highest for kunkers stored at 4 C and then plated on either nonselective (81.3%) or selective (87.5%) media, kunkers stored in ampicillin solution and then plated on selective media (87.5%), and kunkers stored in ampicillin/gentocin solution and plated on selective media (87.5%). Results of this study suggest that optimal isolation rates of P. insidiosum from infected equine tissues are achieved by culturing fresh kunkers on selective media. For samples that cannot be processed immediately, acceptable handling techniques include storage at room temperature for up to 3 days, refrigeration for up to 5 days, shipping on cold packs, and storage in antibiotic solution, each combined with subsequent inoculation on selective media.
Isolation and Multiresidue Detection of Macrolide Endectocides Present in Animal Matrices
Authors: Rudik Irina , Cummings Margie R. , Poppenga Robert H.
Published: May 2011

A simple, rapid, and sensitive method for isolation and detection of macrolide endectocides (moxidectin, doramectin, selamectin, ivermectin, and eprinomectin) in animal sera and liver is described. Fortified sera or homogenized liver samples were treated with sodium chloride followed by organic solvent extraction. No additional steps were required prior to analysis. Separation of analytes and collection of mass information was achieved by liquid chromatography/mass spectrometry with positive atmospheric pressure chemical ionization set in selected ion monitoring mode with each sample analysis complete in 15 minutes. Presence of each compound was confirmed based on 2 separate extracted ion profiles. Detection of avermectins and moxidectin in a range of working standards was achieved at 10, 50, and 100 ppb. Quantitation of these compounds in fortified samples was based on standard calibration curves with R2 > 0.99. Detection limits of 10 ppb for ivermectin, moxidectin, and doramectin, 50 ppb for selamectin, and 100 ppb for eprinomectin were achieved in spiked sera. Recoveries of avermectins and moxidectin in 500 ppb fortified sera fell between 61 and 89% (±5.7–15.7). Analysis of fortified liver gave comparable results with recovery of selamectin of 83–91% ± 18.3. A complete mass spectral fragmentation pattern of selamectin and affordable screening method for 6 macrolide endectocides are reported. Method comparison for salt treatment and solid-phase extraction of fortified samples is discussed.
Evaluation of the Reverse Transcription-Polymerase Chain Reaction/Probe Test of Serum Samples and Immunohistochemistry of Skin Sections for Detection of Acute Bovine Viral Diarrhea Infections
Authors: Ridpath Julia F. , Hietala Sharon K. , Sorden Steve , Neill John D.
Published: May 2011

Bovine viral diarrhea viruses (BVDV) cause both acute and persistent infections. While diagnostic tests have been designed to detect animals persistently infected (PI) with BVDV, the reliability of these tests in detecting acute BVDV infections is not known. It is also possible that acute BVDV infections may be confused with persistent infections in surveys for PI animals. In this study, 2 tests presently in use in diagnostic laboratories to test for PI animals, polymerase chain reaction amplification followed by probe hybridization (RT-PCR/probe) of serum samples and immunohistochemical detection of viral antigen in skin biopsies (IHC), were evaluated for their ability to detect acute BVDV infections. Sixteen colostrum-deprived, BVDV-free, and BVDV-antibody-free calves were infected with 6 different BVDV strains. Clinical signs, seroconversion, and virus isolation indicated that inoculated animals did replicate virus. Virus could be detected in 19% (3/16) of acutely infected animals by the RT-PCR/probe technique. No acutely infected animals were positive by IHC.
Paratuberculosis in a Mandrill (Papio Sphinx)
Authors: Zwick Laura S. , Walsh Timothy F. , Barbiers Robyn , Collins Michael T. , Kinsel Michael J. , Murnane Robert D.
Published: May 2011

A 2.5-year-old captive female mandrill ( Papio sphinx) died following a protracted course of intermittent abdominal bloat, diarrhea, and severe weight loss. Necropsy revealed emaciation and marked gastrointestinal distention with gas and ingesta. Histologic evaluation revealed severe diffuse granulomatous enterocolitis and mesenteric lymphadenitis with massive numbers of 1–2-μm acid-fast bacilli within macrophages. Additionally, there was moderate to severe multifocal myocardial and vascular amyloidosis, moderate multifocal pyogranulomatous interstitial pneumonia with no acid-fast bacteria, and moderate multifocal glossal candidiasis. Samples of feces, ileum, and colon were positive for Mycobacterium avium subsp. paratuberculosis by radiometric culture and a polymerase chain reaction-amplified DNA probe specific for the insertion sequence IS900 of this organism.
Giant Right Atrial Diverticulum in a Foal
Authors: Patterson-Kane Janet C. , Harrison Lenn R.
Published: May 2011

A 5-month-old male Thoroughbred foal with a history of chronic septic arthritis of the tibiotarsal joint and recent respiratory distress was euthanized and a postmortem examination performed. A giant diverticulum communicating with the lateral aspect of the right atrial cavity of the heart was observed. Histologically, the wall was comprised of myocardial tissue containing cavernous vascular spaces. There was gross and histologic evidence of right-sided heart failure. Congenital right atrial diverticula are rare anomalies in humans and have not previously been reported in foals.
Cryptococcosis in a Llama (Lama Glama)
Authors: Bildfell Robert J. , Long Patrick , Sonn Robert
Published: May 2011

Cryptococcosis was diagnosed in a 17-year-old male llama that had been euthanatized following an acute onset of neurologic disease. Tissues affected included the brain, spinal cord, lung, and kidney. The character of the leukocytic response varied from minimal to pyogranulomatous. Cryptococcosis has not been previously reported in a llama, although the infection has been described in 2 other species of New World camelids. The pathogenesis of cryptococcosis is briefly reviewed.
Detection of Rhodococcus Equi by Polymerase Chain Reaction Using Species-Specific Nonproprietary Primers
Authors: Arriaga José Miguel , Cohen Noah D. , Derr James N. , Chaffin M. Keith , Martens Ronald J.
Published: May 2011

Species-specific primers for the polymerase chain reaction (PCR) for the detection of Rhodococcus equi were developed. These primers were based on unique DNA fragments produced from R. equi reference strains and field isolates. Following random amplification of polymorphic DNA from R. equi and R. rhodochrous with a set of 40 arbitrary 10–base pair (bp) primers, a pair of species-specific primers was designed to detect a unique 700-bp fragment of R. equi chromosomal DNA. This PCR product was limited to R. equi and was not detectable in other Rhodococcus species or in a panel of additional gram-positive and gram-negative bacteria.
Development of a Polymerase Chain Reaction and Restriction Typing Assay for the Diagnosis of Bovine Herpesvirus 1, Bovine Herpesvirus 2, and Bovine Herpesvirus 4 Infections
Authors: De-Giuli Luciana , Magnino Simone , Vigo Pier Giorgio , Labalestra Iris , Fabbi Massimo
Published: May 2011

A multiplex polymerase chain reaction (PCR) method coupled with a restriction analysis of PCR products (PCR with restriction fragment length polymorphism) was developed for the simultaneous detection of bovine herpesvirus 1, bovine herpesvirus 2, and bovine herpesvirus 4 infections. The specificity, sensitivity, and practical diagnostic applicability of this method were evaluated. This assay may be also adapted to the diagnosis of suid herpesvirus 1 and equine herpesviruses 1 and 3 and could become a powerful diagnostic tool.