bluuebunny/crossref_metadata_2025 huggingface dataset:
Environmental Air Sampling to Detect Exotic Newcastle Disease Virus in Two California Commercial Poultry Flocks
Authors: Hietala Sharon K. , Hullinger Pamela J. , Crossley Beate M. , Kinde Hailu , Ardans Alex A.
Published: May 2011

The 2002–2003 Exotic Newcastle Disease (END) outbreak in Southern California poultry provided an opportunity to evaluate environmental air sampling as an efficient and cost-effective means of sampling flocks for detection of a circulating virus. Exotic Newcastle Disease virus was detected by real-time reverse transcriptase PCR from air samples collected using a wetted-wall cyclone-style air sampler placed within 2 m of birds in 2 commercial flocks suspected of being naturally exposed to END virus during the outbreak. Exotic Newcastle Disease virus was detected after 2 hours of air sampling the poultry-house environments of the 2 naturally infected flocks.
Canine Renal Pathology Associated with Grape or Raisin Ingestion: 10 Cases
Authors: Morrow Carla M. K. , Valli Victor E. , Volmer Petra A. , Eubig Paul A.
Published: May 2011

Ten dogs suffered acute renal failure after ingesting ≥3 g/kg (dry matter) of grapes or raisins. All dogs had degeneration or necrosis (or both) of proximal renal tubules with basement membranes remaining intact, and epithelial regeneration was observed in 5 out of 10 cases. Mineralized tubular debris or granular to proteinaceous casts (or both) were present in all cases. A golden-brown, globular, intracellular pigment of varying amounts and sizes was observed in 6 out of 10 cases with variable reaction with Prussian blue. Multifocal fibrinous arteritis of the large colon was seen in 2 out of 5 cases with globulin insudation of vessel wall demonstrated by immunohistochemical staining for immunoglobulin (Ig)G and IgM. Mineral analysis on frozen renal tissue from 2 out of 2 cases revealed mildly elevated Ca:P ratio in both. Clinically significant observations were preservation of the integrity of basement membranes after grape-induced tubular injury and presence of early epithelial regeneration. Thus, recovery may be possible if anuria is aggressively managed. With respect to potential pathophysiologic mechanisms, further research into the roles of calcium homeostasis, vascular reactivity, and the significance of the golden-brown pigment is indicated.
Variation among Pathologists in Histologic Grading of Canine Cutaneous Mast Cell Tumors
Authors: Northrup Nicole C. , Harmon Barry G. , Gieger Tracy L. , Brown Cathy A. , Carmichael K. Paige , Garcia Anapatricia , Latimer Kenneth S. , Munday John S. , Rakich Pauline M. , Richey Lauren J. , Stedman Nancy L. , Cheng An-Lin , Howerth Elizabeth W.
Published: May 2011

Ten veterinary pathologists at 1 veterinary institution independently assigned histologic grades to the same 60 canine cutaneous mast cell tumors (MCTs). There was significant variation among pathologists in grading the MCTs ( P < 0.001). The probability of assigning a low grade was significantly higher for the pathologists in this study who use a published reference for histologic grading of canine cutaneous MCTs that allows subcutaneous MCTs or MCTs with mitotic figures to be included in the low-grade category ( P < 0.0001 and P < 0.0001, respectively).
Novel Purification Method for Recombinant 3AB1 Nonstructural Protein of Foot-and-Mouth Disease Virus for Use in Differentiation between Infected and Vaccinated Animals
Authors: Nanni Mariana , Alegre Mariana , Compaired Diego , Taboga Oscar , Fondevila Norberto
Published: May 2011

An indirect enzyme-linked immunosorbent assay (ELISA) was developed for differentiation of animals infected with foot-and-mouth disease virus (FMDV) from vaccinated animals. The test was based on a highly pure and concentrated preparation of recombinant 3AB1 protein obtained by expression in a prokaryotic system, protein separation by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and electro elution. Experimental- and field-serum samples from naive, vaccinated, and infected cattle were tested for anti3AB1 antibody using the ELISA. A cutoff level was set at 35% of the maximum absorbance obtained with a positive control serum (FMDV-infected animal, 21 days postinfection [dpi]). This assay could detect antibodies from sera of animals experimentally infected by contact ( n = 118) with a sensitivity of 97.5%. The specificity was 100%, based on negative test results obtained on 109 sera from naive animals. Remarkably, all sera from animals vaccinated either once ( n = 102) or twice ( n = 30) were negative. In addition, this 3AB1-ELISA could detect seroconversion at 7 dpi in animals inoculated intradermolingually. This assay constitutes an important tool for the rapid detection of FMDV outbreaks in a vaccinated population. In addition, it presents a reliable, economical, and simple method for testing large numbers of serum samples.
A Method for Detecting Complex Vertebral Malformation in Holstein Calves using Polymerase Chain Reaction–Primer Introduced Restriction Analysis
Authors: Kanae Yutaka , Endoh Daiji , Nagahata Hajime , Hayashi Masanobu
Published: May 2011

Complex vertebral malformation (CVM), a hereditary lethal disease in Holstein calves, is characterized by complex anomalies of the vertebral column and limbs in an aborted fetus and in prematurely born, stillborn, and neonatal calves. The mode of inheritance of CVM is autosomal recessive, and CVM is caused by a point mutation from G to T at nucleotide position 559 of the bovine solute carrier family 35 member 3 ( SLC35A3) gene. Although an allele-specific polymerase chain reaction (AS-PCR) is a useful method for diagnosis of CVM, the AS-PCR requires selected DNA polymerases and strictly controlled reaction conditions to obtain reliable results. Therefore, an alternative screening method for the CVM gene would be useful. Polymerase chain reaction–primer introduced restriction analysis (PCR-PIRA) is a method that can be used for detecting a single nucleotide mutation in any gene without a restriction site around the mutation site. In this study, primers were designed to introduce PstI or EcoT22 sites into PCR products from the wild-type and CVM alleles, respectively. The wild-type allele, a heterozygote, and a homozygote of the CVM allele could be discriminated by restriction fragment length polymorphism analysis. Specific introduction of restriction sites into PCR products depending on the change in a single nucleotide of template was shown using a variety of DNA polymerases and PCR machines. Therefore, the PCR-PIRA technique using primers designed in this study might provide a more useful method for extensive screening of CVM.
Gastrointestinal Pythiosis in Two Cats
Authors: Rakich Pauline M. , Grooters Amy M. , Tang Kai-Ning
Published: May 2011

Two young adult male Domestic Shorthair cats living in the southeastern United States were evaluated for signs attributable to partial intestinal obstruction. Physical examination indicated a palpable abdominal mass in each animal. Exploratory laparotomy revealed a large extraluminal mass involving the ileum and mesentery with adjacent mesenteric lymphadenopathy in cat No. 1 and an abscessed mass in the distal duodenum in cat No. 2. Mass resection and intestinal anastomosis were performed in both cats. Histologic evaluation indicated that the intestinal lesions involved primarily the outer smooth muscle layer and serosa and consisted of eosinophilic granulomatous inflammation with multifocal areas of necrosis. In Gomori methenamine silver–stained sections, broad (2.5–7.5 μm), occasionally branching, infrequently septate hyphae were observed within areas of necrosis. A diagnosis of Pythium insidiosum infection was confirmed in both cats by immunoblot serology and by immunoperoxidase staining of tissue sections using a P. insidiosum–specific polyclonal antibody. Cat No. 1 was clinically normal for 4 months after surgery but then died unexpectedly from an unknown cause. Cat No. 2 has been clinically normal for at least 9 months after surgery and appears to be cured on the basis of follow-up enzyme-linked immunosorbent assay serology.
Proliferative Pododermatitis (Canker) with Intralesional Spirochetes in Three Horses
Authors: Nagamine Claude M. , Castro Fernando , Buchanan Ben , Schumacher James , Craig Linden E.
Published: May 2011

Spirochetes were identified histologically in canker (proliferative pododermatitis) in 3 horses. The data suggest that spirochetes may contribute to the pathology of equine chronic foot diseases similar to the manner in which they contribute to the pathology of bovine papillomatous digital dermatitis and contagious ovine digital dermatitis.
Systemic Candidiasis in an Apparently Immunocompetent Dog
Authors: Brown M. Raquel , Thompson Craig A. , Mohamed Fawzi M.
Published: May 2011

Members of the family Candida spp. are ubiquitous dimorphic fungi that normally inhabit the alimentary, upper respiratory, and genital mucosae of mammals. Cell-mediated immunity appears to be an important limitation to the pathologic spread of these fungi. Prolonged immunosuppression, cytotoxic chemotherapy causing neutropenia, diabetes mellitus, long-term glucocorticoid therapy, and prolonged antimicrobial therapy have resulted in an increased incidence of both localized and disseminated candidiasis. This report describes a systemic Candida spp. infection in a dog with no obvious underlying deficiency in host resistance. Cytopathology, histopathology, transmission electron microscopy, and immunohistochemical staining were used to determine the etiology of the causative agent.
Naturally Occurring Eastern Equine Encephalitis in a Hampshire Wether
Authors: Bauer Rudy W. , Gill Marjorie S. , Poston Rob P. , Kim Dae Young
Published: May 2011

Eastern equine encephalitis (EEE) was diagnosed (postmortem) in a sheep with clinical signs attributable to a central nervous system disease. The sheep was febrile and initially had front limb incoordination, which progressed to paralysis of both front and hind limbs during a course of 2 days. The sheep maintained an alert attitude with the ability to eat up to the time of euthanasia. The only clinical pathologic abnormalities were neutrophilia and lymphopenia without appreciable leukocytosis, a moderate hyperglycemia, and an elevated creatine kinase. Treatment included hydrotherapy for lowering body temperature, intravenous fluids, thiamine hydrochloride, tetanus antitoxin, antibiotics, and corticosteroids. The only gross lesion at the time of necropsy was a wet glistening surface of the brain (leptomeninges). Microscopically, there was severe nonsuppurative meningoencephalitis, poliomyelitis, and polyradiculoneuritis with mild multifocal neutrophilic infiltration. The EEE virus was isolated from the brain, and subsequent fluorescent antibody testing for EEE was positive on cell culture.
Acute Depigmentation of Fertile Brown Eggs in a Commercial Layer Operation
Authors: Charlton Bruce R. , Tiwary Asheesh K. , Bickford Arthur A. , Filigenzi Mike
Published: May 2011

Rapid depigmentation of brown eggs is an infrequent but startling event in the commercial egg industry that can result in significant economic losses. Loss of shell pigment in brown-shelled eggs is caused by various factors. In many cases, the exact cause of flock-wide pigment loss remains undetermined. A rapid decline in shell pigmentation was observed in 2 flocks of Hyline brown layers. The lack of evidence of an infectious disease process suggested a feed or management problem. On the basis of a small-scale, “in-house” feeding trial, the feed was identified as the cause of depigmentation. Feed analysis by liquid chromatography with mass spectrometry confirmed the presence of 4,4'-dinitrocarbanilide, a major component of nicarbazin (NCZ). There was no evidence of increased mortality, and only a slight but transient drop in the egg production was observed. Depigmentation effects were rapidly reversed after replacing the feed with NCZ-free feed.
Surveillance of Amyloidosis and other Diseases at Necropsy in Captive Trumpeter Swans (Cygnus Buccinator)
Authors: Meyerholz David K. , Vanloubbeeck Yannick F. , Hostetter Shannon J. , Jordan Dianna M. , Fales-Williams Amanda J.
Published: May 2011

The purpose of this study was to characterize the incidence and diagnostic features of amyloidosis and other diseases found at necropsy in captive trumpeter swans ( Cygnus buccinator). A search of Iowa State University's Department of Veterinary Pathology and Veterinary Diagnostic Laboratory databases yielded 31 trumpeter swan ( C. buccinator) necropsy cases from captive swans in protected habitats. Eleven of the 31 birds had amyloid deposition most commonly in the spleen (8 of 11), liver (7 of 11), and kidney (6 of 11) and less often in the pancreas (2 of 11) and adrenal gland (2 of 11). Amyloid deposition effaced normal tissue with adjacent necrosis and hemorrhage in severe cases. Amyloidosis was most often diagnosed in February and March. Other disease diagnoses in the trumpeter swans included aspergillosis (5 of 31, 16%); bacterial infection (5 of 31, 16%); lead toxicosis (3 of 31, 10%); gout (2 of 31, 6%); parasitic infection (2 of 31, 6%); vitamin E deficiency (1 of 31, 3%); trauma (1 of 31, 3%); and ventricular foreign body (1 of 31, 3%). Histopathologic, toxicologic, and microbiologic analyses did not define an etiologic diagnosis in the deaths of 9 trumpeter swans. In these cases, necropsy lesions included emaciation (5 of 9), enteritis (1 of 9), pulmonary hemorrhage (1 of 9), and no lesions (3 of 9). The number of trumpeter swan case submissions was greatest in January and February. This study provides a reference for veterinary diagnosticians concerning incidence and diagnostic features of amyloidosis and other diseases in captive trumpeter swans of the midwestern United States.
Amplified Fragment Length Polymorphism to Detect Clonal Diversity and Distribution of Mycobacterium Avium Subspecies Paratuberculosis in Selected Minnesota Dairy Cattle
Authors: Kiehnbaum Laura A. , Amonsin Alongkorn , Wells Scott J. , Kapur Vivek
Published: May 2011

The molecular ecology of Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne's disease, is not well understood in the United States. In this study, a DNA fingerprinting method, amplified fragment length polymorphism (AFLP), was used to subtype the pathogen and assess the clonal diversity of MAP in Minnesota dairy herds. Fifty-six fecal culture test–positive isolates from various Minnesota counties and culture dates were analyzed in this study. The AFLP identified 11 profiles with 50% of isolates representing 1 major profile. The major profile was distributed across the state. The genetic diversity of bovine MAP clones in Minnesota based on AFLP analysis of this data appears to be relatively low.
Nictitating Membrane as a Potentially Useful Postmortem Diagnostic Specimen for Classical Swine Fever
Authors: Teifke Jens P. , Lange Elke , Klopfleisch Robert , Kaden Volker
Published: May 2011

The gold standard for diagnosis of classical swine fever (CSF) is cell culture virus isolation combined with reverse transcriptase–polymerase chain reaction (RT-PCR) and fluorescent antibody test (FAT) in cryosections of tonsils, spleen, various lymph nodes, ileum, and kidney. Autolytic and heterolytic samples render correct FAT evaluation difficult and can even yield false-negative or ambiguously positive results. To extend the spectrum of CSF diagnostic specimens, the authors tested whether the nictitating membrane (NM) might be a useful adjunct diagnostic specimen in wild boars and domestic pigs. To accomplish this, results of virus isolation, FAT, and RT-PCR were compared on NM samples and lymphoid tissues, which are the routine specimens of choice for CSF diagnosis. Wild boars ( n = 30) and domestic pigs ( n = 8) were experimentally challenged with various CSF virus (CSFV) strains or isolates of different virulence. The FAT revealed CSFV antigen in surface and tubular adenoid epithelium as well as in lymphatic follicles of the NM. In wild boars and domestic pigs with CSF, a strong agreement was found between results of FAT, virus isolation, and RT-PCR on NM and lymphoid tissues. These results suggest that NM is a useful additional specimen that can provide valuable data for postmortem diagnosis of CSF. The NM is relatively easy to sample at necropsy, and postmortem autolysis and heterolysis of this tissue is minimal compared with internal organs.
Fatal Human Herpesvirus Type 1 Infection in a White-Handed Gibbon (Hylobates Lar)
Authors: Landolfi Jennifer A. , Wellehan James F. X. , Johnson April J. , Kinsel Michael J.
Published: May 2011

This report documents a case of spontaneous, fatal, and likely recrudescent human herpesvirus type 1 (HHV-1) infection in a captive white-handed gibbon ( Hylobates lar) confirmed by polymerase chain reaction (PCR). An approximately 44-year-old, captive, female, white-handed gibbon with a history of recurrent conjunctivitis and occasional seizures became acutely weak, disoriented, and ataxic. A postictal state was suspected by caretakers and veterinary staff, and euthanasia was ultimately elected because of lack of clinical improvement with supportive care. No significant abnormalities were detected at necropsy. Histologically, sections of cerebrum and midbrain contained minimal to mild, multifocal lymphoplasmacytic meningoencephalitis with numerous intranuclear viral inclusions within astrocytes and some neurons. The presumptive diagnosis of HHV-1–induced encephalitis was strengthened by nested PCR amplification of a segment of the herpesvirus DNA polymerase gene. Sequences from this region have been found to be unique to each herpesvirus species, thus identifying HHV-1 as the likely etiologic agent in this case. Positive HHV-1 serology from several years before the terminal episode suggested that the disease was most likely due to recrudescence of latent HHV-1 infection.
The Adequacy of Sample Type/Weight and Incubation Period on Detection of Salmonella Spp. in Slaughter Cattle
Authors: Vimont A. J. M. , Kiermeier A. , Padula D. J. , Holds G. L. , Pointon A. M.
Published: May 2011

The purpose of this study was to examine the adequacy of different sample types (fecal and rumen content), rumen-content sample weight (1, 10, and 25 g), and incubation period on the detection of Salmonella spp. in grass-fed beef cattle at slaughter. The culture technique was the same for all samples and followed the Australian Standard (AS 1766.2.5—1991). Sample adequacy was defined as the ratio between the overall prevalence, as obtained from samples identified as positive by any sample type/weight, and the estimated prevalence, as obtained from samples identified as positive by a particular sample type/weight. Sample adequacy reflects the likelihood of a sample of a particular type and weight to contain the organism of interest and hence is related to the sensitivity of the diagnostic test. It was found that sample adequacy differed between sample types and weights: 37.5% for both a 10-g fecal sample and a 1-g rumen sample, 77.1% for a 10-g rumen sample, and 79.2% for a 25-g rumen sample. On this basis, it is strongly recommended that sample type and weight be considered in the design of studies that aim to quantify Salmonella contamination in cattle.
Effects of Delayed or Prolonged Fixation on Immunohistochemical Detection of Bovine Viral Diarrhea Virus Type I in Skin of Two Persistently Infected Calves
Authors: Miller Margaret A. , Ramos-Vara José A. , Kleiboeker Steven B. , Larson Robert L.
Published: May 2011

The effects of delayed or prolonged fixation on immunohistochemical detection of bovine viral diarrhea virus (BVDV) antigen were evaluated in skin. Ear-notch specimens from 2 calves persistently infected with BVDV type 1 were handled in 1 of 3 ways: 1) fixed in formalin promptly and processed for immunohistochemistry (IHC) after 3–176 days; 2) held at 3–4°C in plastic bags up to 10 days, then fixed in formalin for 2–5 days before processing; or 3) exposed to room air and temperature for 1–5 days before formalin fixation. Immunohistochemical staining intensity was evaluated without the knowledge of specimen handling. Staining of specimens that had been promptly fixed in formalin was moderate to strong at all fixation periods through 36 days, weak or no staining was evident in specimens fixed for 176 days. Refrigerated specimens typically had moderate to strong immunohistochemical staining. Even after 10 days of refrigeration before fixation, all immunohistochemical reactions were positive. However, no immunohistochemical staining was detected in any specimen that was exposed to room air. Results indicate that prompt formalin fixation is optimal for BVDV IHC. Samples can be held in formalin at least 36 days, without loss of reactivity. A 1-day delay in fixation caused no loss of reactivity, provided the specimen was refrigerated and protected from desiccation.
Comparison of Two Enzyme-Linked Immunosorbent Assays for Diagnosis of Mycobacterium Avium Subsp. Paratuberculosis
Authors: McKenna S. L. B. , Sockett D. C. , Keefe G. P. , McClure J. , VanLeeuwen J. A. , Barkema H. W.
Published: May 2011

Enzyme-linked immunosorbent assays (ELISAs) are used in Johne's disease (JD) control programs as a first screening for presence of the disease in a herd. A high sensitivity of the ELISA is therefore important, yet the commonly used ELISAs have relatively low sensitivity. The inclusion of an absorption phase, although improving specificity, potentially decreases sensitivity. Sera and feces of 383 adult dairy cows in 8 herds were used to compare the test characteristics of an absorbed and a nonabsorbed indirect ELISA for the detection of JD. The absorbed ELISA is based on a protoplasmic antigen, whereas the nonabsorbed uses a lipoarabinomannan-based antigen. The potential advantage of the nonabsorbed ELISA is that it may be less specific and more sensitive. Two herds certified free of JD were used to compare the specificity of the ELISAs. The other herds used to compare sensitivity were either infected with Mycobacterium avium subsp. paratuberculosis or had unknown status. Using fecal culture as a gold standard, the diagnostic specificity for the absorbed and nonabsorbed ELISAs were 98.4% and 87.9%, respectively. The diagnostic sensitivity was 72.4% and 65.5% for the absorbed and the nonabsorbed ELISA, respectively. Furthermore, a comparison using a fecal DNA probe as the comparison standard resulted in both ELISAs having a sensitivity of 61.9%. Agreement between the 2 ELISAs was moderate, with a kappa statistic of 0.58. The nonabsorbed ELISA did not have a higher sensitivity and had a lower specificity than the absorbed ELISA. Therefore, in this population, there was no advantage gained with using the nonabsorbed ELISA.
A Simple and Rapid Chromatographic Strip Test for Detection of Antibody to Porcine Reproductive and Respiratory Syndrome Virus
Authors: Lyoo Y. S. , Kleiboeker S. B. , Jang K.-Y. , Shin N. K. , Kang J.-M. , Kim C.-H. , Lee S.-J. , Sur J.-H.
Published: May 2011

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be a major economic problem for swine industries worldwide despite several disease-reduction strategies such as age-segregated early weaning and all-in-all-out pig movement. Routine diagnosis of PRRSV is carried out by the combined use of an antibody-detecting enzyme-linked immunosorbent assay (ELISA), immunofluorescence, reverse transcription-polymerase chain reaction, and virus isolation. These assays require specialized laboratory equipment in addition to multistep sample handling and sample preparation. The objective of this study was to evaluate a simple pen-side assay (BioSign™ PRRSV) for rapid detection of PRRSV antibody based on a lateral flow chromatographic strip immunoassay system. This assay uses Escherichia coli–expressed viral nucleocapsid protein antigen for detecting antibodies against PRRSV in swine sera. In this report, the authors describe the evaluation of this assay using sera from both clinical samples and experimentally infected piglets. The results were compared with those of a standard, commercially available antibody ELISA (HerdChek®PRRS ELISA) and an indirect immunofluorescence assay using the same serum samples. The BioSign™ PRRSV assay was capable of detecting antibodies in sera known to contain antibodies to PRRSV, resulting in 93.2% sensitivity for samples from experimentally infected pigs and 98.7% sensitivity for clinical serum samples. For sera that did not contain antibodies to PRRSV, the specificity was found to be 98.5% and 99.2% for clinical and experimental serum samples, respectively.
Lymphoid Follicular Cloacal Inflammation Associated with a Novel Herpesvirus in Juvenile Alligators (Alligator Mississippiensis)
Authors: Govett Pamela D. , Harms Craig A. , Johnson April J. , Latimer Kenneth S. , Wellehan James F. X. , Fatzinger Michael H. , Christian L. Shane , Kelly Terra R. , Lewbart Gregory A.
Published: May 2011

Multifocal hyperemic nodules and plaques associated with the cloacal mucosa of juvenile alligators ( Alligator mississippiensis) at a public aquarium were investigated. Grossly, pale pink to dark red multifocal, circular lesions of varying degrees of severity were identified on the cloacal and, in males, phallus mucosa. Cloacal mucosa biopsies were obtained from 2 of the alligators. These samples were examined histologically and by polymerase chain reaction (PCR) using consensus primers targeting a conserved region of the herpesvirus polymerase gene. Microscopically, the lesions were characterized as submucosal lymphoid follicles with hyperemia and hemorrhage. No inclusion bodies were observed. Minimal to no anisokaryosis was present, and no etiologic agents were identified. Through PCR, a band consistent in size with herpesvirus was observed. Tissues showing similar clinical, histopathologic, and PCR findings were collected from animals at an alligator farm several months later. Sequencing of the PCR amplicon resulted in a 180-base pair sequence that shared 85% sequence identity with tortoise herpesvirus-1.
Segmental Aplasia of Uterine Body in an Adult Mixed Breed Dog
Authors: Oh Ki-Seok , Son Chang-Ho , Kim Bang-Sil , Hwang Shun-Shin , Kim You-Jung , Park Su-Jin , Jeong Jae-Ho , Jeong Cheol , Park Sung-Hee , Cho Kyoung-Oh
Published: May 2011

Segmental aplasia of the uterine body was diagnosed in a 5-year-old, mixed breed bitch. Abdominal radiography and transabdominal ultrasonography revealed marked dilation of fluid-filled uterine horns with no evidence of a uterine body. Sex hormone assays did not detect the presence of estradiol-17 (β; however, progesterone (2 ng/ml) was found in the serum, indicating anestrus. On gross examination of the reproductive tract, the uterine body was absent, apparently never formed. In its place, a cord-like piece of tissue was identified as an aplastic/dysplastic remnant, connecting the cervix and right uterine horn. The tip of the cord-like piece branched into 5 string-like pieces of tissue, 1 of which was connected to the region dividing the left and right uterine horns. Both the uterine horns were dilated markedly revealing hydrometra. Histologically, uterine body remnant tissues from the endometrium, myometrium, and perimetrium were detected in proximal and distal parts of the uterine body. The string-like piece of tissue connecting the uterine body remnant and the uterine horn consisted of a round cluster of smooth muscle cells surrounding a central core of adipose tissue with blood vessels. It was concluded that the hydrometra observed in both uterine horns was induced by an obstruction resulting from segmental aplasia in the uterine body. This is the first known report of segmental aplasia in the uterine body of a bitch.
Spontaneous Aortic Dissecting Hematoma in Two Dogs
Authors: Boulineau Theresa Marie , Andrews-Jones Lydia , Van Alstine William
Published: May 2011

This report describes 2 cases of spontaneous aortic dissecting hematoma in young Border Collie and Border Collie crossbred dogs. Histology was performed in one of the cases involving an unusual splitting of the elastin present within the wall of the aorta, consistent with elastin dysplasia as described in Marfan syndrome in humans. The first case involved a young purebred Border Collie that died suddenly and the second case involved a Border Collie crossbred dog that died after a 1-month history of seizures. Gross lesions included pericardial tamponade with dissection of the ascending aorta in the former case and thoracic cavity hemorrhage, mediastinal hematoma, and aortic dissection in the latter. Histologic lesions in the case of the Border Collie crossbred dog included a dissecting hematoma of the ascending aorta with elastin dysplasia and right axillary arterial intimal proliferation.
Status Spongiosus of White Matter in Newborn Gelbvieh-Cross Calves
Authors: O'Toole D. , Montgomery D. L. , Steadman L. , O'Rourke B. , Russell W. , Dennis J.
Published: May 2011

Various forms of status spongiosus occur in neonatal cattle, the best characterized of which is due to mutations of the branched-chain alpha-keto acid dehydrogenase complex (BCKD), resulting in bovine maple syrup urine disease (MSUD, branched-chain ketoaciduria). A distinctive neurological syndrome was identified between 1998 and 2003 in 9 calves in a 250-cow stabilized Gelbvieh-Red Angus herd. Both sexes were affected (6 heifers, 3 bulls), with a low annual incidence (3 cases in 1998; no cases in 1999; 2 cases in 2000; 2 in 2001; 1 in 2002; 1 in 2003). Affected calves were born full-term, unable to stand, and had constant whole-body tremors when stimulated. Animals remained in lateral recumbency until death or euthanasia; the longest survival time was 10 days postpartum. The principal histological change in 2 affected calves was diffuse, moderately severe bilaterally symmetrical status spongiosus with Alzheimer type II cells throughout the white matter of the brain. Myelin deficits were not evident and vacuoles were due to cleaved myelin sheaths. Neither recognized mutation of MSUD was identified in the E1a subunit of BCKD in 2 affected calves, 8 dams that gave birth to affected calves, a grand-dam of 3 affected calves, or a sire of 1 calf. Amino acid analysis of serum from 1 affected calf revealed normal concentrations of branched-chain amino acids, indicating that this disease is distinct from MSUD. The genetic and biochemical basis for the disorder, provisionally named congenital status spongiosus of Gelbvieh-cross cattle, is undetermined. The pattern of inheritance was not established.
Effect of Egg Yolk on the Detection of Mycobacterium Avium Subsp. Paratuberculosis Using the ESP II Liquid Culture System
Authors: Harris N. Beth , Robbe-Austerman Suelee , Payeur Janet B.
Published: May 2011

Rapid diagnosis of paratuberculosis in infected cattle is important for the successful control of Johne disease within herds. Thus, improving culture methods for Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) will aid in the identification of asymptomatic animals. Egg yolk is a component of the media used for growing M. paratuberculosis, but its requirement as a supplement has not been reported. Using the ESP II liquid culture system, 2 different sources and 5 concentrations (3.3%, 1.6%, 0.8%, 0.4%, and 0%) of egg yolk were analyzed. Egg yolk source did not affect either recovery rate or time to detection, but both parameters were significantly improved when the 3.3% egg yolk concentrations (final volume) were used over media containing no egg yolk. This study also assessed the recovery of M. paratuberculosis from fecal samples that were cultured multiple times using Herrold egg yolk agar (HEY). Specimens containing greater than 70 cfu/g feces could routinely be identified as positive for M. paratuberculosis after only 1 culture attempt, whereas specimens with fewer bacteria were only intermittently positive, even after 5 replicate cultures. Therefore, this study indicates that the sensitivity of the Trek Diagnostic ESP II liquid culture system for M. paratuberculosis is affected by egg yolk concentration and that single culture attempts using HEY solid media may not identify specimens containing low numbers of bacteria.
Variation among Pathologists in the Histologic Grading of Canine Cutaneous Mast Cell Tumors with Uniform Use of a Single Grading Reference
Authors: Northrup N. C. , Howerth E. W. , Harmon B. G. , Brown C. A. , Carmicheal K. P. , Garcia A. P. , Latimer K. S. , Munday J. S. , Rakich P. M. , Richey L. J. , Stedman N. L. , Gieger T. L.
Published: May 2011

Ten veterinary pathologists independently assigned histologic grades to the same 60 canine cutaneous mast cell tumors using the Patnaik classifications. The degree of agreement in grading among the pathologists was compared with the degree of agreement among the same pathologists in a previous study, in which each pathologist used the reference for grading that he/she uses routinely. Mean agreement improved significantly from 50.3% to 62.1% with uniform use of the Patnaik classifications ( P = 0.00001), suggesting that there is value in uniform application of a single grading scheme for canine cutaneous mast cell tumors. Agreement among pathologists was still not 100%, suggesting that a more objective grading scheme should be developed and that other histologic indicators of prognosis should be investigated.
Cloning and Sequencing of Equine Cardiac Troponin I and Confirmation of Its Usefulness as a Target Analyte for Commercial Troponin I Analyzers
Authors: Rishniw Mark , Simpson Kenny W.
Published: May 2011

The analysis of cardiac troponin I (cTnI) in the diagnosis of myocardial injury in domestic animals is gaining popularity. In this study, equine cTnI was sequenced and compared with previously characterized cTnI from other species. A 6-amino-acid N-terminal deletion unique to the horse was identified. This deletion was outside the epitope region of cTnI recognized by most commercial immunoassays and did not affect the ability of a commercial analyzer system to detect recombinant equine cTnI. No function could be ascribed to the deleted portion. These data support the use of commercial analyzers in measuring equine cTnI.
A Method for Extracting Genomic DNA from Individual Elaphostrongyline (Nematoda: Protostrongylidae) Larvae and Differentiation of Elaphostrongylus Spp. from Parelaphostrongylus Spp. by PCR Assay
Authors: Chilton Neil B. , Huby-Chilton Florence , Lankester Murray W. , Gajadhar Alvin A.
Published: May 2011

This article reports a rapid and effective method for the extraction and purification of genomic DNA (gDNA) from individual first-stage larvae (L1) of elaphostrongyline nematodes that had been stored frozen or fixed in 95% ethanol for 1 to 5 years. The method was highly effective for L1s of all 6 species of elaphostrongylines, based on polymerase chain reaction (PCR) amplification of a partial fragment of the first internal transcribed spacer (ITS-1) of the ribosomal DNA. Differences were detected in the sizes of partial ITS-1 amplicons between the 2 elaphostrongyline genera, Elaphostrongylus and Parelaphostrongylus. The reliability of the ITS-1 PCR assay was tested by using L1s of unknown identity from Newfoundland and Labrador, Canada. The ability to consistently isolate gDNA from individual L1s, together with a simple PCR-based method to distinguish between Parelaphostrongylus and Elaphostrongylus, have important implications for diagnostic testing and for conducting epizootiological studies on these parasites of veterinary importance.
Seasonal Meningoencephalitis in Holstein Cattle Caused by Naegleria Fowleri
Authors: Daft Barbara M. , Visvesvara Govinda S. , Read Deryck H. , Kinde Hailu , Uzal Francisco A. , Manzer Michael D.
Published: May 2011

Primary amoebic meningoencephalitis is a fulminant infection of the human central nervous system caused by Naegleria fowleri, a free-living amoeba that thrives in artificially or naturally heated water. The infection usually is acquired while bathing or swimming in such waters. The portal of entry is the olfactory neuroepithelium. This report describes fatal meningoencephalitis caused by N. fowleri in Holstein cattle that consumed untreated surface water in an area of California where summer temperatures at times exceed 42°C. In the summers of 1998 and 1999, severe multifocal necrosuppurative hemorrhagic meningoencephalitis was observed in brain samples from nine 10-20-month-old heifers with clinical histories of acute central nervous system disease. Olfactory lobes and cerebella were most severely affected. Lesions were also evident in periventricular and submeningeal neuropil as well as olfactory nerves. Naegleria fowleri was demonstrated by immunohistochemistry in brain and olfactory nerve lesions and was isolated from one brain. Even though cultures of drinking water did not yield N. fowleri, drinking water was the likely source of the amoeba. The disease in cattle closely resembles primary amoebic meningoencephalitis in humans. Naegleria meningoencephalitis should be included among differential diagnoses of central nervous system disease in cattle during the summer season in areas with high ambient temperatures.
Gossypol Toxicosis in a Dog Consequent to Ingestion of Cottonseed Bedding
Authors: Uzal Francisco A. , Puschner Birgit , Tahara John M. , Nordhausen Robert W.
Published: May 2011

Six dogs died after accidental ingestion of cottonseed bedding. No clinical signs of illness were observed prior to death. A full diagnostic workup was performed on one of these dogs. At necropsy, the lungs were congested and edematous, and the liver was firm, congested, and had a marked reticular pattern. There was also moderate ascites. Histopathologic examination revealed multifocal myocardial degeneration and necrosis, severe pulmonary edema, and chronic passive congestion of the lungs, heart, liver, and kidneys. Transmission electron microscopy of the myocardium revealed disruption of myofibrils, chromatin condensation, and disrupted and swollen mitochondria. The cottonseed bedding contained 1,600 mg/kg of free gossypol, a concentration considered toxic for monogastric animals. The stomach content revealed the presence of gossypol, thus confirming ingestion of cottonseed. Gossypol poisoning in dogs is extremely rare and has not yet been associated with cottonseed bedding. This first documented case of gossypol poisoning in a dog, caused by the ingestion of cottonseed bedding, demonstrates how specific toxicological analysis is crucial in reaching an accurate diagnosis.
Histochemical and Immunohistochemical Evidence of a Bacterium Associated with Lesions of Epizootic Bovine Abortion
Authors: Anderson M. L. , Kennedy P. C. , Blanchard M. T. , Barbano L. , Chiu P. , Walker R. L. , Manzer M. , Hall M. R. , King D. P. , Stott J. L.
Published: May 2011

Epizootic bovine abortion (EBA), a tick-transmitted disease of pregnant cattle grazing foothill pastures, is a major cause of reproductive failure in California and adjacent states. Affected fetuses develop a chronic disease, resulting in late-term abortion or premature calving. Despite investigations spanning 50 years, to the authors' knowledge, the etiologic agent of EBA has not yet been isolated from affected fetuses or the tick vector. The diagnosis of EBA is based on gross and microscopic lesions. Recently, documentation that the etiologic agent is susceptible to antibiotics and identification of a unique 16S deltaproteobacterial rDNA gene sequence in 90% of thymus tissues from aborted fetuses have supported the role of a bacterial infection as the cause of EBA. To determine whether bacteria could be detected in the tissues, histochemical staining and immunohistochemical procedures were used on formalin-fixed, paraffin-embedded tissues. Use of a modified Steiner silver stain revealed small numbers of intracytoplasmic bacterial rods in 37 of 42 thymic samples from EBA-affected fetuses. Improved detection was achieved by use of immunohistochemical staining with serum from EBA-affected fetuses that resulted in detection of numerous bacterial rods in the cytoplasm of histiocytic cells in the thymus from all 42 EBA-affected fetuses. Immunohistochemical examination of additional tissues from 21 field and experimental EBA cases revealed positively stained intracytoplasmic bacterial rods in many organs with inflammatory lesions. Use of the modified Steiner stain and immunohistochemical staining of tissues from negative-control fetuses failed to reveal organisms. To the authors' knowledge, this is the first report to document morphologic evidence of a bacterium associated with the lesions of EBA.
Detection of Canine Parvovirus in Fecal Samples using Loop-Mediated Isothermal Amplification
Authors: Kang Jong-Il , Park Nam-Yong , Cho Ho-Seong
Published: May 2011

Loop-mediated isothermal amplification (LAMP) is a novel, sensitive, and rapid technique for detection of genomic DNA. The end-product of the technique is a white precipitate of magnesium pyrophosphate that is visible without the use of gel electrophoresis. The LAMP method was applied to the detection of canine parvovirus (CPV) genomic DNA. A set of 4 primers, 2 outer and 2 inner, were designed from CPV genomic DNA targeting the VP2 gene. The optimal reaction time and temperature for LAMP were determined to be 60 minutes and 63°C. On the basis of results for 50 canine fecal samples using polymerase chain reaction (PCR) analysis as the gold standard, the relative sensitivity of LAMP was 100% and the relative specificity was 76.9%. The detection limit of the LAMP method was 10−1 median tissue culture infective doses (TCID50)/ml, compared with 10 TCID50/ml for PCR analysis. In addition to the advantage resulting from visual detection of the end product, the LAMP method is very rapid, requiring only 1 hour to complete. This assay would be a viable alterative to PCR analysis for diagnosis of CPV infection in dogs. The LAMP method holds promise for use as a diagnostic assay for CPV detection in a clinical setting.
Detection of Foot-and-Mouth Disease Virus: Comparative Diagnostic Sensitivity of Two Independent Real-Time Reverse Transcription-Polymerase Chain Reaction Assays
Authors: King Donald P. , Ferris Nigel P. , Shaw Andrew E. , Reid Scott M. , Hutchings Geoff H. , Giuffre Angelica C. , Robida John M. , Callahan Johnny D. , Nelson William M. , Beckham Tammy R.
Published: May 2011

Rapid and accurate diagnosis is central to the effective control of foot-and-mouth disease (FMD). It is now recognized that reverse-transcription polymerase chain reaction (RT-PCR) assays can play an important role in the routine detection of FMD virus (FMDV) in clinical samples. The aim of this study was to compare the ability of 2 independent real-time RT-PCR (rRT-PCR) assays targeting the 5′ untranslated region (5′UTR) and RNA polymerase (3D) to detect FMDV in clinical samples. There was concordance between the results generated by the 2 assays for 88.1% (347 of 394) of RNA samples extracted from suspensions of epithelial tissue obtained from suspect FMD cases. The comparison between the 2 tests highlighted 19 FMDV isolates (13 for the 5′UTR and 6 for the 3D assay), which failed to produce a signal in 1 assay but gave a positive signal in the other. The sequence of the genomic targets of selected isolates highlighted nucleotide substitutions in the primer or probe regions, thereby providing an explanation for negative results generated in the rRT-PCR assays. These data illustrate the importance of the continuous monitoring of circulating FMDV field strains to ensure the design of the rRT-PCR assay remains fit for purpose and suggest that the use of multiple diagnostic targets could further enhance the sensitivity of molecular methods for the detection of FMDV
Use of Reverse Transcription-Polymerase Chain Reaction to Determine the Stability of Rabies Virus Genome in Brains Kept at Room Temperature
Authors: Rojas Anaya Edith , Loza-Rubio Elizabeth , Banda Ruiz Víctor Manuel , Hernández Baumgarten Eliseo
Published: May 2011

In tropical and subtropical climates, the shipment of animal brains for rabies diagnosis may be a problem because brain specimens sometimes arrive decomposed at the diagnostic laboratory. In this situation, reverse transcription-polymerase chain reaction (RT-PCR) may serve as a potential solution because of its high sensitivity. However, little is known about the stability of rabies viral RNA in decomposed brain tissue. To determine the stability of rabies virus genomic RNA in brain samples, 72 mice were inoculated with the challenge virus strain-11 of rabies virus. After incubation period, mice were euthanized to obtain their brains. These were categorized in 2 different groups. In the first group, 36 brains were kept at room temperature (25–27°C) immediately after euthanasia. In the second group, the other 36 inoculated brains were frozen at −70°C and later maintained at room temperature. In both groups, RT-PCR was performed at days 0, 1, 2, 3, 4, 7, 10, 12, 16, 18, 23, and 26 by using primers previously described in the literature and a primer set specifically designed for a Mexican variant of vampire-bat rabies. Reverse-transcriptase PCR experiments were performed in 3 different inoculated brains, in which the direct fluorescent antibody (DFA) test was previously conducted to detect rabies viral antigen in the brains kept at room temperature and in the frozen brains. The DFA test resulted positive in both groups up to day 7. In brain samples stored at ambient temperature (25–27°C), the intensity of the RT-PCR band started to diminish by day 12; however, rabies virus genome could be successfully amplified by RT-PCR up to 23 days. These results indicate that brain samples kept at ambient temperature (up to 27°C) may reach a reference laboratory in an adequate state for rabies diagnosis by RT-PCR.
Efficient Screening of the Cystinuria-Related C663T Slc3a1 Nonsense Mutation in Newfoundland Dogs by Denaturing High-Performance Liquid Chromatography
Authors: Matos Augusto José Ferreira , Mascarenhas Cláudia , Magalhães Paula , Pinto Jorge Pereira
Published: May 2011

Cystinuria in Newfoundland dogs is a metabolic disease associated with a nonsense mutation in the exon 2 of the Slc3a1 gene. Similar to type I human cystinuria, heterozygote carriers are not affected by the disease and do not reveal differences in urinary concentration of dibasic amino acids when compared with normal dogs. However, through a recessive mode of inheritance, these dogs are able to transmit the disease to their offspring. Early detection of mutation carriers through cost-effective reliable methods is therefore essential for the implementation of breeding methods aimed at the eradication of the disease. Denaturing high-performance liquid chromatography (DHPLC) is a recently developed technique for rapid and efficient screening of nucleotide polymorphisms in polymerase chain reaction–amplified products. This technique was used for the identification of the C663T Slc3a1 mutation in Portuguese Newfoundland dogs. Polymerase chain reaction products amplified from a region containing the C663T locus were subjected to DHPLC analysis, and results were double checked by DNA sequencing. Results showed the presence of the mutation in 6 of the 22 dogs tested. Urine biochemical parameters correlated well with the number of mutated Slc3a1 copies, and homozygotes for the C663T mutation were the only dogs diagnosed with cystinuria. Sequence analysis confirmed the DHPLC results, demonstrating that the technique could be a reliable alternative to sequencing for the rapid and cost-effective identification of mutations in canine breeds.