bluuebunny/crossref_metadata_2025 huggingface dataset:
Development of Real-Time Polymerase Chain Reaction Assays for Rapid Detection and Differentiation of Wild-Type Pseudorabies and Gene-Deleted Vaccine Viruses
Authors: Ma Wenjun , Lager Kelly M. , Richt Juergen A. , Stoffregen William C. , Zhou Fanghong , Yoon Kyoung-Jin
Published: May 2011

The successful eradication of pseudorabies in U.S. domestic swine was accomplished through the use of glycoprotein E (gE) deleted modified live virus vaccines and an accompanying gE differential enzyme-linked immunosorbent assay (ELISA). Yet, pseudorabies virus (PRV) was established in feral swine in the United States, becoming a potential reservoir of PRV for infection of domestic swine and other native wildlife. A critical need for the current PRV surveillance program in the United States is the rapid detection of PRV infection. For this reason, a set of 2 real-time polymerase chain reaction (PCR) assays by using TaqMan chemistry was developed and evaluated for their capability in the detection and differentiation of field and vaccine strains of PRV. PCR primers and probes were designed for gB and gE genes of PRV, respectively. The newly developed PRV-specific real-time PCR assays could detect all wild-type PRV isolates from diagnostic submissions and differentiate them from vaccine strains. The analytical sensitivity of the assays was approximately 0.1 plaque-forming units per reaction. The assays were highly specific for PRV, because no positive results were obtained from testing other common swine viral pathogens and other animal herpesviruses. The results of testing samples from domestic and feral swine and from bovine showed that the real-time PCR assays are more sensitive than gel-based PCR. These results demonstrated the potential application of the developed real-time PCR assays as a differential test for rapid and specific detection of PRV in domestic and feral swine, as well as nonporcine species that can be infected with PRV and serve as carriers.
Rapid Detection and Differentiation of Wild-Type and Three Attenuated Lapinized Vaccine Strains of Classical Swine Fever Virus by Reverse Transcription Polymerase Chain Reaction
Authors: Pan Chu-Hsiang , Jong Ming-Hwa , Huang Yu-Liang , Huang Tien-Shine , Chao Parn-Hwa , Lai Shiow-Suey
Published: May 2011

A simple one-step reverse transcription polymerase chain reaction (RT-PCR) method was developed based on T-rich insertions in the viral genome for simultaneous detection and differentiation of wild type and vaccine strains of Classical swine fever virus (CSFV). The CSFV-specific primers were designed to contain the sequences of the T-rich insertion sites that exist uniquely in the 3' nontranslated regions (3′ NTR) of the genome of lapinized CSFV vaccine strains. By using a one-step RT-PCR or a nested PCR followed by an agarose gel electrophoresis or a multicapillary electrophoresis, the wild-type and lapinized vaccine strains of CSFV in clinical samples could be detected and accurately distinguished. These assays can be applied to at least 3 attenuated lapinized vaccine strains, lapinized Philippines Coronel (LPC), hog cholera lapinized virus (HCLV), and Chinese strain (C strain). The detection limit of the wild-type virus was 6.3 TCID50 (50% tissue culture infective dose)/ml for RT-PCR and 0.63 TCID50/ml for nested PCR. In previous studies, notable T-rich insertions of 12–13 nucleotides (nt) were found in the 3′ NTR of the genome of lapinized vaccine strains of CSFV. However, this study discovered that 2 T-rich insertions, 42 and 36 nt in length, are present in the viral genome of lapinized vaccine strains LPC/PRK (primary rabbit kidney) and LPC/TS (Tam-Sui), respectively. These T-rich insertions of 12, 36, and 42 nt length increases the size of PCR fragments, which are favorable genetic markers for rapid detection of and differentiation between wild-type and different lapinized vaccine strains of CSFV.
Lack of Correlation between Antibody Titers to Fibrinogen-Binding Protein of Streptococcus Equi and Persistent Carriers of Strangles
Authors: Davidson Ann , Traub-Dargatz Josie L. , Magnuson Roberta , Hill Ashley , Irwin Vivienne , Newton Richard , Waller Andrew , Smith Kenneth , Callan Robert J. , Meehan Mary , Owen Peter , Salman Mo
Published: May 2011

Previously published studies have neither used nor reported the results of an indirect enzyme-linked immunosorbent assay (iELISA) to measure serologic responses in natural outbreaks of strangles. The concept of using serologic responses to identify persistent carriers of Streptococcus equi has been proposed but not scientifically evaluated. The specific aims of the current study were to determine the duration and level of truncated fibrinogen-binding protein-specific (SeM allele 1) antibody production in ponies involved in a natural outbreak of strangles and to determine if test results from this serologic iELISA could predict persistent carrier status. Serologic samples were obtained before and after an outbreak of naturally occurring strangles infection. Persistent carriers of S. equi were identified via culture and polymerase chain reaction (PCR) testing of lavage fluid collected from the guttural pouches and nasopharynx or swabs of the nasopharynx after recovery from acute disease and at postmortem examination. Logistic regression analysis was used to determine if an association existed between serologic response and persistent carrier state. The ELISA reported in the current study definitively confirmed a recent exposure to S. equi. However, the measured serologic response did not predict carrier status in this strangles outbreak. Therefore, a guttural-pouch endoscopy with subsequent culture or PCR testing to detect S. equi remains the most accurate method available for the identification of persistent carriers.
Development of a Novel Biochip for Rapid Multiplex Detection of Seven Mastitis-Causing Pathogens in Bovine Milk Samples
Authors: Lee Kuo-Hua , Lee Jai-Wei , Wang Shih-Wen , Liu Lu-Yuan , Lee Mei-Fen , Chuang Shih-Te , Shy Yih-Min , Chang Chu-Li , Wu Ming-Che , Chi Chau-Hwa
Published: May 2011

To efficiently prevent and treat bovine mastitis and minimize its effect on the dairy industry, a sensitive, rapid, and specific test is required for identifying the mastitis-causing pathogens. In this study, a biochip capable of detecting 7 common species of mastitis-causing pathogens, including Corynebacterium bovis, Mycoplasma bovis, Staphylococcus aureus, and the Streptococcus spp. S. agalactiae, S. bovis, S. dysgalactiae, and S. uberis, within 6 hr was developed. The technique is based on DNA amplification of genes specific to the target pathogens and consists of 4 basic steps: DNA extraction of bacteria, polymerase chain reaction, DNA hybridization, and colorimetric reaction. To examine the accuracy and specificity of this biochip, a preliminary test with 82 random quarter milk samples were analyzed and compared with results from conventional microbiological methods conducted simultaneously. Results from all but 1 sample analyzed by the biochip were in agreement with those analyzed by bacteriology. The biochip could be a feasible tool for rapidly diagnosing mastitis-causing pathogens in milk and providing information for a more effective treatment to cure mastitis.
Interaction of Ionophore and Vitamin E in Knockdown Syndrome of Turkeys
Authors: Crespo Rocio , Shivaprasad H. L. , Sommer Franz , Puschner Birgit
Published: May 2011

Monensin and vitamin E concentrations, as well as histopathology of skeletal muscles and myocardium, were evaluated in broad-breasted white turkeys kept in commercial facilities. Turkeys with knockdown syndrome had myopathy of skeletal muscles, but no lesions in the myocardium. Generally, concentration of monensin in serum was highest in turkeys diagnosed with knockdown syndrome given more than 90 mg/kg of monensin in the diet, followed by turkeys diagnosed with knockdown syndrome given <90 mg/kg of monensin in the diet, healthy turkeys fed a diet that contained <90 mg/kg of monensin, and finally healthy turkeys fed a diet free of monensin (not detectable). However, the concentration of monensin was highly variable within each group, and the median was lower than the average. Vitamin E concentrations in the livers varied from low–normal to below normal and were statistically higher in healthy turkeys fed a diet free of monensin than in the livers of birds from the 3 groups exposed to monensin. This suggests that the concentration of monensin in serum positively correlates to the severity of clinical signs and pathology and to the amount of monensin in the feed. Although the methodology developed to detect serum monensin concentrations is beneficial and accurate for case investigations, it is recommended that several samples from each flock be evaluated because of variation within a flock. The current study also suggests that monensin in the feed could induce lower concentrations of vitamin E in the liver of turkeys and can predispose the turkeys to knockdown syndrome.
Antimicrobial Resistance in Salmonella Enterica Subspecies Enterica Serovar Dublin from Dairy Source Calves in the Central San Joaquin Valley, California (1998–2002)
Authors: Berge Anna Catharina B. , Thornburg Elizabeth , Adaska John M. , Moeller Robert B. , Blanchard Patricia C.
Published: May 2011

The present study describes antimicrobial resistance patterns of Salmonella enterica subspecies enterica serovar Dublin ( S. Dublin) in clinical submissions from calves and temporal and farm-type trends in antimicrobial resistance patterns of the isolates. A total of 300 isolates of S. Dublin were obtained from fecal or internal organs of calves fewer than 120 days of age originating from 84 dairies and 18 calf ranches from July 1998 to December 2002. The isolates were susceptibility tested to a panel of 10 antimicrobials using the disk diffusion assay. Temporal and farm-type trends in individual antimicrobial inhibition zone sizes were assessed and antimicrobial resistance patterns were described using cluster analysis. Isolates obtained from calf ranches compared with dairies exhibited decreased susceptibility to florfenicol, gentamicin, neomycin, sulfisoxazole, sulfamethoxazole/trimethoprim, and tetracycline. During the years 1998–2002, decreasing susceptibility was seen for ceftiofur, enrofloxacin, and sulfamethoxazole/trimethoprim. There were 20 different antimicrobial resistance patterns in the isolate set, indicating that S. Dublin has the ability to transfer and pick up resistance genes with relative ease. The trends seen in antimicrobial resistance in S. Dublin may likely be linked to antimicrobial drug use in young calves.
Mycobacterium Fortuitum Infection Interference with Mycobacterium Bovis Diagnostics: Natural Infection Cases and a Pilot Experimental Infection
Authors: Michel Anita L.
Published: May 2011

Mycobacterium fortuitum and at least 1 unidentified species of soil mycobacteria were isolated from lymph nodes from 4 of 5 African buffalo ( Syncerus caffer) that had been culled because of positive test results using the Bovigam assay. The buffalo were part of a group of 16 free-ranging buffalo captured in the far north of the Kruger National Park (South Africa) assumed to be free of bovine tuberculosis. No Mycobacterium bovis was isolated. To investigate the possible cause of the apparent false-positive diagnosis, the Mycobacterium isolates were inoculated into 4 experimental cattle and their immune responses monitored over a 13-week period, using the gamma interferon assay. The immune reactivity was predominantly directed toward avian tuberculin purified protein derivative (PPD) and lasted for approximately 8 weeks. During that period 3 of 4 cattle yielded positive test results on 1 or 2 occasions. The immune responsiveness was boosted when the inoculations were repeated after 15 weeks, which led to 2 subsequent positive reactions in the experimental animal that did not react previously. Including an additional stimulatory antigen, sensitin prepared from M. fortuitum in the gamma interferon assay, showed that it was able to elicit a detectable gamma interferon response in all 4 experimentally inoculated cattle when applied in parallel with bovine and avian tuberculin PPD for the stimulation of blood samples. The implications of occasional cross-reactive responses in natural cases of infection with environmental mycobacteria in the diagnosis of bovine tuberculosis in African buffalo and cattle in South Africa are discussed.
Tularemia in Range Sheep: An Overlooked Syndrome?
Authors: O'Toole Donal , Williams Elizabeth S. , Woods Leslie W. , Mills Kenneth , Boerger-Fields Amy , Montgomery Donald L. , Jaeger Paula , Edwards William H. , Christensen Dwayne , Marlatt William
Published: May 2011

Abortion and death caused by Francisella tularensis were well recognized in range flocks of domestic sheep in Idaho, Montana, and Wyoming in the first 6 decades of the 20th century. The current report describes 4 episodes of tularemia in 3 range flocks in Wyoming and South Dakota in 1997 and 2007 (1 flock was affected twice). Flock owners reported that ticks were unusually numerous and commonly present on sheep during outbreaks. Tularemia presented as late-term abortions (3 episodes) or listlessness and death in lambs and, to a lesser extent, ewes (1 episode). Lesions were multifocal pinpoint necrotic foci in tissues, particularly spleen, liver, and lung. An immunohistochemical procedure demonstrated F. tularensis, particularly in necrotic foci. The diagnosis was corroborated by bacterial isolation and, in individual cases, by serology, fluorescent antibody assay, and/or polymerase chain reaction detection of F. tularensis. Diagnosticians in endemic areas should include tularemia as a differential diagnosis when investigating late-term abortions or outbreaks of fatal illness in young lambs, particularly in years of high tick activity and when characteristic necrotic foci occur in spleen, liver, and lung.
Equine Giant Cell Tumor of Soft Parts: A Series of 21 Cases (2000–2007)
Authors: Bush Jamie M. , Powers Barbara E.
Published: May 2011

In horses, giant-cell tumors of soft parts are rare neoplasms, with the majority of reported cases occurring within the hind limb muscles and soft tissues in older horses. The following article documents 21 cases of equine giant-cell tumors of soft parts clinically examined within the state of Colorado from 2000 to 2007. The majority of cases occurred in male horses aged 10 years or older. Nine (43%) arose within the hind limbs. Key histologic features included numerous multinucleated giant cells and hemosiderin-laden macrophages admixed with a spindle-cell proliferation. The majority demonstrated liposarcomatous change, variable areas of necrosis and hemorrhage, and an intermediate number of mitotic figures. Immunohistochemical results demonstrated 2 distinct cell populations: vimentin-expressing neoplastic mesenchymal cells and CD18 (histiocytic marker) expressing multinucleated giant cells. These results suggest a mesenchymal origin of the neoplasm with possible recruitment of the secondary histiocytic population. Surgical excision was attempted in the majority of horses and was considered clinically complete. A recurrence of the neoplasm was documented in 1 horse and 1 mule. In 18 horses, surgical excision, regardless of margin integrity, appeared successful with no recurrence of disease documented. Unfortunately, 10 of 21 horses were lost to follow-up within approximately 3 months of surgery. Of the 11 remaining horses that were available for follow-up evaluation, there has been no evidence of metastasis. A larger case series with more controlled follow-up is necessary to evaluate malignant potential and the importance of complete surgical excision.
Renal Mucus Gland Cystadenomas in a Horse
Authors: Loynachan Alan T. , Bryant Uneeda K. , Williams Neil M.
Published: May 2011

A 35-year-old horse was submitted to the necropsy service at the University of Kentucky Livestock Disease Diagnostic Center. At necropsy, multiple 1–4-cm-diameter cystic structures were incidentally identified unilaterally in the right renal medulla and the cortex. On histologic examination, the cystic structures compressed the normal renal architecture, were lined by tall columnar epithelium that formed occasional papillary projections, and contained large amounts of mucicarmine and periodic acid–Schiff-positive mucinous material. The masses were diagnosed as renal mucus-gland cystadenomas. This tumor should be considered as a differential diagnosis when cystic structures are identified in the equine kidney.
Acalculous and Clostridial Cholecystitis in a Pig
Authors: Starost Matthew F. , Burkholder Tanya H.
Published: May 2011

A 21-month-old domestic Hanford pig ( Sus scrofa domestica) in a 1-year study for experimental myocardial infarction was euthanized at the end of the study. One week earlier, the animal had symptoms and elevated clinical chemistry results suggestive of hepatobiliary disease, which resolved after medical therapy. At necropsy, the gallbladder was markedly enlarged, discolored, and had a thickened wall. Within the gallbladder, there was abundant friable green-brown material. A culture of the gallbladder luminal material yielded Clostridium perfringens type A. Histopathology of the gallbladder demonstrated multifocal areas of necrosis of varying depths, admixed with an inflammatory infiltrate that was also observed on the serosa and within the associated adipose tissue. Luminal material was composed of cellular debris and bile sludge admixed with numerous bacterial rods. Smooth-muscle hypertrophy of numerous small arterioles with narrowed lumina was observed in the gallbladder. A diagnosis of acalculous cholecystitis presumably because of ischemia of the gallbladder with secondary clostridial infection was made. To the authors' knowledge, this is the first reported case of acalculous cholecystitis with evidence of vascular compromise in a pig, as well as cholecystitis secondarily attributed to C. perfringens type A.
A Chromatographic Strip Test for Rapid Detection of One Lineage of the H5 Subtype of Highly Pathogenic Avian Influenza
Authors: Cui Shangjin , Tong Guangzhi
Published: May 2011

A rapid immune colloidal gold test strip for detecting 1 lineage of the H5 subtype of highly pathogenic avian influenza was developed based on membrane chromatography with monoclonal antibodies (mAbs) against hemagglutinin (HA) of the H5 subtype of Avian influenza virus (AIV). Bagg albino C mice that were immunized with AIV A/Goose/Guangdong/3/96(H5N1) yielded 4 hybridomas secreting mAbs (1A6, 1B8, 3C4, and 8F6) specific for HA. Monoclonal antibody 3C4 was conjugated with colloidal gold as the detector antibody. On the test strip, 1A6, in combination with rabbit anti-H5N1 polyclonal antibody, was used as the capture complex at the “test line,” and goat antimouse immunoglobulin G antibody was used as the capture antibody at the “control line.” The strip test correctly detected the H5 subtype of AIV in known positive samples and did not react with the standard antigens of AIV H1-H4 or H6-H14, or the antigens of Newcastle disease virus, Infectious bursal disease virus, Infectious bronchitis virus, or Avian infectious laryngotracheitis virus. When 483 clinical specimens suspected of infection with H5N1 were tested, results from the strip test were nearly identical with those from the HA/hemagglutination inhibition test, reverse transcription polymerase chain reaction, and a commercial enzyme-linked immunosorbent assay kit. Because the newly developed strip was specific, rapid, and sensitive, it should be useful for diagnosing the H5 subtype of AIV and for investigating its epidemiology. The test strip is only reliable for the lineage of H5N1. As for the other strains of AIV, further experiments will be needed.
Acute Phase Protein Response in Goats
Authors: González Félix H. D. , Tecles Fernando , Martínez-Subiela Silvia , Tvarijonaviciute Asta , Soler Laura , Cerón José J.
Published: May 2011

Acute phase proteins (APPs) are important diagnostic indicators of inflammatory disturbances in animals. The objectives of the current study were to validate analytical methods for measuring haptoglobin (Hp), serum amyloid A (SAA), acid soluble glycoprotein (ASG), fibrinogen, and albumin concentrations in goats and to evaluate their response to an inflammatory stimulus in this species. Intra- and interassay coefficients of variation (CVs) were in the range 0.07–9.31% and 1.83–12.68%, respectively, for all APPs and showed good precision. All assays determined APPs in a linear manner at different sample dilutions with high correlation coefficients with the exception of fibrinogen, which was measured by the heat precipitation method. Subcutaneous injection of turpentine oil induced an increase in Hp, SAA, ASG, and fibrinogen serum concentrations and a decrease in albumin concentration.
Evaluation of three Methods for Measurement of Hemoglobin and Calculated Hemoglobin Variables with the ADVIA 120® and ADVIA 2120® Systems in Goats
Authors: Bauer Natali , Moritz Andreas
Published: May 2011

The present study investigated 3 methods of hemoglobin (Hb) determination in goats using the ADVIA 120 and ADVIA 2120 systems. Ethylenediaminetetraacetic acid anticoagulated caprine blood samples (n = 40 goats) were subjected to Hb determination via the cyanmethemoglobin methods in both instruments and a novel, cyanide-free, colorimetric method with the ADVIA 2120. Statistical analysis of the data included a linear regression, Passing-Bablok regression, and Bland-Altman diagram. Colorimetric Hb results determined with both analyzers had excellent correlation ( r = 0.98); however, a mean proportional bias of −19.1% was present in comparison to the reference method. There also was excellent agreement between cellular Hb concentrations when measured with both analyzers ( r = 0.96), and the constant bias was close to zero. However, imprecision was higher compared to colorimetric methods. Excellent to fair agreement was evident for all calculated erythrocyte and Hb variables. Because of the excellent correlation between the ADVIA 120 and ADVIA 2120, the cyanide-free method of Hb determination could be used with caprine blood specimens; however, the proportional bias must be considered.
Determination of Cardiac Troponin I in the Blood and Heart of Calves with Foot-and-Mouth Disease
Authors: Tunca Recai , Sozmen Mahmut , Erdogan Hidayet , Citil Mehmet , Uzlu Erdogan , Ozen Hasan , Gokçe Erhan
Published: May 2011

The current study was designed to determine the changes of the cardiac troponin I (cTnI) expression in blood and tissue during the myocardial degeneration in calves with foot-and-mouth disease (FMD). Seventeen crossbred calves presenting pathological signs for FMD confirmed by viral analysis were studied. A biochemistry panel and immunohistochemistry were performed on 17 diseased calves and 7 calves used as controls. Creatine kinase (CK), CK-myocardial band (CK-MB), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) activities were analyzed for both groups. Cardiac troponin I levels were measured by a commercially available enzyme-linked immunosorbent assay kit. Mean cTnI (14.8 ± 1.9 ng/ml) concentration and CK (573 ± 407 U/l), CK-MB (238 ± 37 U/l), AST (84 ± 7), and LDH (298 ± 29 U/l) activities were higher in FMD cases compared with controls. Immunohistochemistry revealed loss or depletion of cTnI expression in myocardium of all cases. None of the 7 controls showed loss of cTnI expression. Increased serum cTnI concentration correlated with myocardial injury and loss of cTnI immunolabeling in cardiomyocytes of calves with FMD.
Duplex Real-Time Polymerase Chain Reaction for Simultaneous Detection and Quantification of Anaplasma Marginale and Anaplasma Centrale
Authors: Decaro Nicola , Carelli Grazia , Lorusso Eleonora , Lucente Maria Stella , Greco Grazia , Lorusso Alessio , Radogna Arianna , Ceci Luigi , Buonavoglia Canio
Published: May 2011

Anaplasma marginale and Anaplasma centrale are rickettsial pathogens responsible for acute disease and mild infections, respectively, in cattle herds. A duplex real-time polymerase chain reaction (PCR) assay with probes labeled with different fluorophores was developed for simultaneous detection and quantification of A. marginale and A. centrale DNA in bovine blood samples. The assay was able to detect as few as 101 and 102 DNA copies for A. marginale and A. centrale, respectively, with optimal specificity and reproducibility. Analysis by real-time and nested PCR carried out on 54 samples previously tested by reverse line blot hybridization showed that the established duplex real-time PCR assay can detect and quantify the 2 Anaplasma spp., even if present simultaneously in the same blood samples. Such an assay could be used in pathogenesis studies on bovine acute anaplasmosis.
Serological Evaluation of Precolostral Serum Samples to Detect Bovine Viral Diarrhea Virus Infections in Large Commercial Dairy Herds
Authors: Schefers Jeremy , Munoz-Zanzi Claudia , Collins James E. , Goyal Sagar M. , Ames Trevor R.
Published: May 2011

The authors propose that screening newborn calves for Bovine viral diarrhea virus (BVDV) antibody prior to colostrum feeding is a useful strategy to detect herds with endemic BVDV infection. In the current study, precolostral serum samples of newborn calves in 2 Minnesota and 2 California dairy farms were examined. Precolostral BVDV antibodies were detected by serum neutralization and enzyme-linked immunosorbent assay in 7.4% (33/446) and 6.2% (32/515) of newborn calves in the California and Minnesota herds, respectively. The serum samples were also tested by reverse transcription polymerase chain reaction (RT-PCR), and BVDV was detected in 1.6% (7/446) and 3.5% (18/515) of newborn calves in the California and Minnesota herds, respectively. The primary advantages of precolostral testing are that calves congenitally infected with BVDV and seropositive at birth represent a larger percentage of calves born than BVDV-viremic calves and that fewer animals would need to be tested with an antibody test than a RT-PCR or antigen detection test to detect endemic BVDV infections at the herd level. Testing for BVDV antibody in calves prior to colostrum feeding detects fetal infections in both late-gestating cows and nonlactating heifers. Precolostral serum antibody detection is not confounded by vaccination and may be a more sensitive screening method than bulk milk RT-PCR and nonvaccinated sentinel calf strategies in large dairy herds.
Canine Procalcitonin Messenger RNA Expression
Authors: Kuzi Sharon , Aroch Itamar , Peleg Keren , Karnieli Ohad , Klement Eyal , Dank Gillian
Published: May 2011

Procalcitonin is considered an acute phase protein used as both a marker of infection and prognosis in human medicine. Canine procalcitonin has been previously sequenced; however, its use as a diagnostic or prognostic tool in dogs has never been assessed. A quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay for canine procalcitonin messenger RNA (mRNA) was developed. Whole blood samples were collected from ill and healthy dogs. RNA was extracted and the realtime PCR was assessed. The patients' diagnoses, complete blood cell count, and differential leukocyte count results were recorded. Based on the diagnosis, dogs were divided into 5 groups: inflammatory, infectious, neoplastic, other diseases, and healthy controls. Procalcitonin mRNA expression and the hematological measures were compared between groups, and their correlations were assessed. Procalcitonin mRNA expression was assessed in 70 dogs, including infectious (17), noninfectious inflammatory (17), neoplastic (18), other diseases (7), and healthy controls (11), and was significantly ( P < 0.001) higher in all ill dogs versus controls. Procalcitonin may therefore be considered an acutephase protein in dogs. However, there were no significant differences in procalcitonin mRNA expression between ill dog groups and no correlations between its expression levels and hematological measures. In 5 dogs of all disease categories, procalcitonin mRNA expression was measured twice during the course of disease. The changes in its levels were in agreement with the clinical evaluation of improvement or deterioration, suggesting a possible prognostic value.
Diagnostic Significance of Serum Glycated Albumin in Diabetic Dogs
Authors: Sako Toshinori , Mori Akihiro , Lee Peter , Takahashi Tomoya , Izawa Takehito , Karasawa Sino , Furuuchi Miho , Azakami Daigo , Mizukoshi Mina , Mizutani Hisashi , Kiyosawa Yoshio , Arai Toshiro
Published: May 2011

Measurements of serum fructosamine, glycated hemoglobin, and glycated albumin (GA) are increasingly used to complement serum glucose concentration for better management of diabetes mellitus. Fructosamine tests are currently not performed in veterinary medicine in Japan. As such, the measurement of GA may serve as a replacement test. Therefore, in the current study, serum GA and fructosamine were evaluated for a positive correlation in dogs, and, depending on the correlation, a reference range of GA percentage would also be determined from healthy control dogs. The degree of glycemic control in diabetic dogs was determined by fructosamine concentration. A positive correlation between GA and fructosamine was observed with both normal and diabetic animals. In addition, the reference interval of serum GA percentage in control dogs was determined to be 11.4–11.9% (95% confidence interval). Interestingly, no significant difference in serum GA percentages was observed between samples from diabetic dogs with excellent glycemic control and control dogs. However, good, fair, and poor glycemic control diabetic dogs resulted in a significant increase in serum GA percentages in comparison with control dogs. These results suggest that serum GA may be a useful diagnostic indicator, substituting for fructosamine, to monitor glycemic control in diabetic dogs.
Evaluation of Four DNA Extraction Methods for the Detection of Tritrichomonas Foetus in Feline Stool Specimens by Polymerase Chain Reaction
Authors: Stauffer Stephen H. , Birkenheuer Adam J. , Levy Michael G. , Marr Henry , Gookin Jody L.
Published: May 2011

Feces are increasingly valued as practical samples for molecular diagnosis of infectious disease. However, extraction of polymerase chain reaction (PCR) quality DNA from fecal samples can be challenging because of coextraction of PCR inhibitors. Because the type and quantity of PCR inhibitors is influenced by diet, endogenous flora, and concurrent disease, it is unlikely that extraction method performance with human feces can be directly extrapolated to that of domestic cats. In the present study, 4 commercially available DNA extraction methods were examined for their influence on the sensitivity of PCR for the detection of Tritrichomonas foetus in feline stool. DNA was extracted from serially diluted feline-origin T. foetus trophozoites in the absence or presence of feline feces. The ZR Fecal DNA kit was identified as affording the greatest analytical sensitivity and reproducibility and was able to detect ≥10 T. foetus organisms per 100 mg feces in 100% of PCR reactions. Further, the identified extraction method could be completed in the shortest time of all kits tested.
Occipitoatlantoaxial Malformation in an Adult Goat
Authors: Seva Juan I. , Gómez Serafin , Pallarés Francisco J. , Sánchez Pedro , Bernabé Antonio
Published: May 2011

An occipitoatlantoaxial malformation was diagnosed in a 1-year-old Murciano-Granadina goat. At clinical examination, the head and cranial part of the neck were deviated to the right. Clinical signs of spinal cord or brain disease were not observed. At necropsy, morphological abnormalities were seen in the craniovertebral junction and cervical vertebrae, characterized by a firm attachment and incomplete articulation between the occipital bone and the atlas, and scoliosis in the cervical regions. The definitive diagnosis was bilateral asymmetrical occipitoatlantoaxial fusion with rotation of the atlas and atlantoaxial subluxation. To the authors' knowledge, this case report is the second occipitoatlantoaxial malformation described in a goat and the first description in an adult goat.
Diffuse Intestinal T-cell Lymphosarcoma in a Yellow-Naped Amazon Parrot (Amazona Ochrocephala Auropalliata)
Authors: Souza Marcy J. , Newman Shelley J. , Greenacre Cheryl B. , Avenell James S. , Wall Jonathan S. , Phillips Jeffrey C. , Fry Michael M. , Donnell Robert L. , Daniel Gregory B.
Published: May 2011

A 10-year-old, intact, female yellow-naped Amazon parrot ( Amazona ochrocephala auropalliata) was examined because of anemia, lymphocytic leukocytosis, regurgitation, and weight loss. A positive fecal occult blood and monoclonal globulinopathy were present. A distended proventriculus and diffusely thickened loops of small intestine with irregular luminal surfaces were identified with contrast radiography and contrast computed tomography. A micro positron emission tomography scan was performed with 18F-fluorodeoxyglucose. Diffuse intestinal T-cell lymphosarcoma was diagnosed based on histopathology and immunohistochemistry of full thickness small intestinal biopsies. The patient was treated with a multidrug chemotherapy protocol with little to no effect. Euthanasia was elected, and intestinal lymphosarcoma was confirmed on histopathology of necropsy intestinal samples; no other organs demonstrated neoplastic infiltration. To the authors' knowledge, no reports are currently available detailing the clinical presentation or diagnosis of diffuse intestinal T-cell lymphosarcoma in any avian species.
Lipid-Rich Pleural Mesothelioma in a Dog
Authors: Avakian Arlen , Alroy Joseph , Rozanski Elizabeth , Keating John , Rosenberg Andrew
Published: May 2011

An 11-year-old, neutered, male Golden Retriever cross dog was euthanized following a history of recurrent pericardial effusions. At necropsy, blood-tinged pericardial and intrathoracic effusions were seen along with numerous firm to hard plaque-like masses that studded the epicardial, pericardial, mediastinal, and costal pleural surfaces. Within the right thorax, the lesions coalesced into a large mass that occupied most of the cavity. Histologically, the masses were composed of solid sheets and papillary aggregates of medium-sized polygonal cells that contained abundant vacuolated to clear cytoplasm. Some of the cytoplasmic vacuoles stained positive with oil red O. The stroma contained metaplastic trabeculae of woven and lamellar bone. Immunohistochemically, the neoplastic cells expressed vimentin, pancytokeratin, and S-100 protein. Transmission electron microscopy corroborated the presence of intracytoplasmic vacuoles and demonstrated prominent intercellular junctional complexes and apically located microvilli. These findings are consistent with a lipid-rich variant of mesothelioma. To the authors' knowledge, this is the first report of a lipid-rich mesothelioma in a dog.
Ulcerative Enterocolitis in Two Goats Associated with Enterotoxin- and beta2 Toxin–Positive Clostridium Perfringens Type D
Authors: Uzal Francisco A. , Fisher Derek J. , Saputo Juliann , Sayeed Sameera , McClane Bruce A. , Songer Glenn , Trinh Hien T. , Miyakawa Fernandez Mariano E. , Gard Sharon
Published: May 2011

Enterotoxemia caused by Clostridium perfringens type D in sheep is believed to result from the action of epsilon toxin (ETX). However, the sole role of ETX in the intestinal changes of the acute and chronic forms of enterotoxemia in goats remains controversial, and the synergistic action of other C. perfringens toxins has been suggested previously. The current study examined 2 goats that were found dead without premonitory clinical signs. Gross lesions at necropsy consisted of multifocal fibrinonecrotic enterocolitis, edematous lungs, and excess pleural fluid. Histologically, there were multifocal fibrinonecrotic and ulcerative ileitis and colitis, edema of the colonic serosa, and proteinaceous interstitial edema of the lungs. Clostridium perfringens type D carrying the genes for enterotoxin (CPE) and beta2 toxin (CPB2) was cultured from intestinal content and feces of 1 of 2 goats, while C. perfringens type D CPB2–positive was isolated from the other animal. When multiple colonies of the primary isolations from both animals were tested by Western blot, most of the isolates expressed CPB2, and only a few isolates from the first case expressed CPE. Alpha toxin and ETX were detected in ileal and colonic contents and feces of both animals by antigen capture enzyme-linked immunosorbent assay. CPB2, but not CPE, was identified in the small and large intestines of both goats by immunohistochemistry. These findings indicate that CPB2 may have contributed to the necrotic changes observed in the intestine, possibly assisting ETX transit across the intestinal mucosa.
Mixed Urolith (Struvite and Calcium Oxalate) in a Ferret (Mustela Putorius Furo)
Authors: Del Angel-Caraza Javier , Chávez-Moreno Orlando , García-Navarro Sergio , Pérez-García Carlos
Published: May 2011

Urolithiasis has been reported in veterinary literature in some species of the Mustelidae family, including ferrets. In this species, the most common form of urolithiasis is that caused by struvite. The current study examined the case of mixed urolith in an 8-month-old female ferret ( Mustela putorius furo) with antecedents of strangury and hematuria. A cystotomy was carried out to remove the urolith, which had a rough surface and a homogeneously porous interior and was formed by a mixture of struvite (60%) and calcium oxalate dehydrated (40%).
Fat Embolism Secondary to Yellow Fat Disease in an Appaloosa Horse
Authors: Suárez-Bonnet Alejandro , de los Monteros Antonio Espinosa , Herráez Pedro , Rodríguez Francisco , Andrada Marisa , Caballero Maria José
Published: May 2011

A 4-year-old female Appaloosa horse was referred to the Department of Comparative Pathology, Veterinary Faculty, with a history of colic and acute respiratory distress. At necropsy, gross lesions consisted of extensive firm, multinodular, hemorrhagic foci in fat tissues with yellow-brown discoloration. The most affected areas were peritoneal fat and perirenal, epicardial, and subcutaneous adipose tissues. Other findings were hepatic lipidosis and multiple 1–1.5 cm hemorrhagic foci scattered in both lungs. Histopathological examination revealed severe degeneration and necrosis of adipose tissue with dystrophic calcification. The necrotic fat was infiltrated by abundant foamy macrophages and multinucleated giant cells. Based on these histopathological changes, yellow fat disease, also called nutritional panniculitis, was diagnosed. In addition, the microscopic examination of lung and kidney sections stained with osmium tetroxide and oil red O revealed numerous lipid droplets within glomerular and alveolar septal capillaries. To the authors' knowledge, this is the first description of the development of fat embolism in horses affected by yellow fat disease.
Isolation of Arcanobacterium Hippocoleae from a Case of Placentitis and Stillbirth in a Mare
Authors: Bemis David A. , Bryant Mary Jean , Kania Stephen A. , Newman Shelley J.
Published: May 2011

Based on the scarcity of reports in veterinary medicine literature, it appears that Arcanobacterium spp. are rarely isolated from horses. Recently, a single isolate from vaginal discharge in a mare was characterized as a new species, Arcanobacterium hippocoleae. The present report describes a case of necrosuppurative placentitis and stillbirth in an American Quarterhorse mare. Numerous colonies of irregular, Gram-positive, rod-shaped bacteria were observed by histological examination within fibrin lattice associated with placental lesions. Arcanobacterium hippocoleae was isolated in large numbers from the placenta, lung, and stomach contents. To the authors' knowledge, this is the first report of placentitis associated with Arcanobacterium spp. in a horse and the third reported isolation of A. hippocoleae from a horse.
Idiopathic Arterial Medial Calcification of the Thoracic Arteries in an Adult Horse
Authors: Fales-Williams Amanda , Sponseller Beatrice , Flaherty Heather
Published: May 2011

A 6-year-old, gelded, Paint horse displayed clinical signs of muscle wasting and limb stiffness for a 6-month period. The horse's clinical signs abated with corticosteroid therapy, but returned upon cessation of treatment. Upon necropsy, severe lesions of aortic thickening and aortic valve rigidity were observed. Histologically, the tunica media of the aorta, coronary arteries, and pulmonary arteries were expanded by foci of elastin fiber calcification and extracellular matrix with lacunae formation. The vascular lesions are comparative to what has been described as medial arterial calcification, seen in humans suffering from chronic renal failure or diabetes mellitus. No exposure to vitamin D-containing plants or feedstuff could be documented at the time of onset or during the period of clinical signs. The current case describes dramatic lesions of arterial medial calcification of the aorta, coronary, and pulmonary arteries of undetermined cause.
Routine Phenotypic Identification of Bacterial Species of the Family Pasteurellaceae Isolated from Animals
Authors: Dousse Florence , Thomann Andreas , Brodard Isabelle , Korczak Bozena M. , Schlatter Yvonne , Kuhnert Peter , Miserez Raymond , Frey Joachim
Published: May 2011

Pasteurellaceae are bacteria with an important role as primary or opportunistic, mainly respiratory, pathogens in domestic and wild animals. Some species of Pasteurellaceae cause severe diseases with high economic losses in commercial animal husbandry and are of great diagnostic concern. Because of new data on the phylogeny of Pasteurellaceae, their taxonomy has recently been revised profoundly, thus requiring an improved phenotypic differentiation procedure to identify the individual species of this family. A new and simplified procedure to identify species of Actinobacillus, Avibacterium, Gallibacterium, Haemophilus, Mannheimia, Nicoletella, and Pasteurella, which are most commonly isolated from clinical samples of diseased animals in veterinary diagnostic laboratories, is presented in the current study. The identification procedure was evaluated with 40 type and reference strains and with 267 strains from routine diagnostic analysis of various animal species, including 28 different bacterial species. Type, reference, and field strains were analyzed by 16S ribosomal RNA ( rrs) and rpoB gene sequencing for unambiguous species determination as a basis to evaluate the phenotypic differentiation schema. Primary phenotypic differentiation is based on β-nicotinamide adenine dinucleotide (β-NAD) dependence and hemolysis, which are readily determined on the isolation medium. The procedure divides the 28 species into 4 groups for which particular biochemical reactions were chosen to identify the bacterial species. The phenotypic identification procedure allowed researchers to determine the species of 240 out of 267 field strains. The procedure is an easy and cost-effective system for the rapid identification of species of the Pasteurellaceae family isolated from clinical specimens of animals.
Diagnostic Performance of Assays for the Detection of Anti-Porcine Reproductive and Respiratory Syndrome Virus Antibodies in Serum and Muscle Transudate (“Meat Juice”) Based on Samples Collected under Experimental Conditions
Authors: Molina Ramon M. , Chittick Wayne , Nelson Eric A. , Christopher-Hennings Jane , Rowland Raymond R. R. , Zimmerman Jeffrey J.
Published: May 2011

Three assays were evaluated for their ability to detect antibodies against Porcine reproductive and respiratory syndrome virus (PRRSV) in porcine muscle transudate (“meat juice”) samples. Samples were derived from 91 pigs inoculated with PRRSV isolate VR-2332 and 46 age-matched controls. Serum and muscle ( Musculus longissimus dorsi) samples were collected from randomly selected animals euthanized at ∼14-day intervals from 28 to 202 days postinoculation. Serum samples were assayed at a dilution of 1:40, and muscle transudate samples were assayed at 5 dilutions (1:2, 1:5, 1:10, 1:20, 1:40) using a commercial PRRSV antibody enzyme-linked immunosorbent assay (ELISA). In addition, muscle transudate samples were tested using an indirect fluorescent antibody test (IFAT) at 5 dilutions (1:2, 1:5, 1:10, 1:20, 1:40). Attempts to assay muscle transudate samples for neutralizing antibodies using a modified fluorescent focus neutralization assay were unsuccessful. Receiver operator characteristic (ROC) curve analyses were used to estimate cutoff thresholds and the associated diagnostic sensitivities and specificities for ELISA and IFAT at each dilution. For ELISA, muscle transudate samples at the ROC-optimized cutoffs were >95% sensitive and 100% specific at each dilution. At a cutoff dilution of ≥1:5, the IFAT diagnostic sensitivity and specificity of muscle transudate was estimated at 63.3% and 100%, respectively. These findings validated the use of muscle transudate samples in PRRSV surveillance programs based on ELISA antibody testing.
Evidence for Persistent Bovine Viral Diarrhea Virus Infection in a Captive Mountain Goat (Oreamnos Americanus)
Authors: Nelson Danielle D. , Dark Michael J. , Bradway Daniel S. , Ridpath Julia F. , Call Neill , Haruna Julius , Rurangirwa Fred R. , Evermann James F.
Published: May 2011

Bovine viral diarrhea (BVD) viruses are pestiviruses that have been isolated from domestic and wild ruminants. There is serologic evidence of pestiviral infection in more than 40 species of free-range and captive mammals. Vertical transmission can produce persistently infected animals that are immunotolerant to the infecting strain of Bovine viral diarrhea virus (BVDV) and shed virus throughout their lives. Seven species (white-tailed deer, mouse deer, eland, domestic cattle, alpaca, sheep, and pigs) have been definitively identified as persistently infected with BVDV. This study provides serological, molecular, immunohistochemical, and histological evidence for BVDV infection in 2 captive mountain goats from a zoological park in Idaho. The study was triggered by isolation of BVDV from tissues and immunohistochemical identification of viral antigen within lesions of a 7-month-old male mountain goat (goat 1). Blood was collected from other mountain goats and white-tailed and mule deer on the premises for BVDV serum neutralization, viral isolation, and reverse transcription polymerase chain reaction. One 3-month-old mountain goat (goat 2) was antibody negative and BVDV positive in serum samples collected 3 months apart. This goat subsequently died, and though still antibody negative, BVDV was isolated from tissues and identified by immunohistochemistry within lesions. Sequencing and phylogenetic analysis identified the isolates as BVDV-2. These findings provide evidence of persistent infection in a mountain goat, underscoring the need for pestivirus control strategies for wild ruminants in zoological collections.