bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Multiple Sampling on Diagnostic Sensitivity
Authors: Thurmond Mark C. , Johnson Wesley O.
Published: May 2011

Overall diagnostic sensitivity is the probability that a diagnostic procedure will detect an agent if the tested animal is indeed infected. The overall or effective sensitivity is a function of both the probability that the assay will detect the agent if it is present in the sample tested and the probability that the agent will be present in the sample tested if the animal is infected with the agent. Thus, even with a highly sensitive assay, the probability of detecting an infected animal may be low or nil if the sampling procedure failed to capture the agent in samples tested by the assay. In this article, it is demonstrated how increased frequency of testing, such as testing multiple subsamples, can have a profound effect on increasing the overall sensitivity of a diagnostic procedure.
Detection of Three Avian Respiratory Viruses by Single-Tube Multiplex Reverse Transcription–Polymerase Chain Reaction Assay
Authors: Malik Yashpal S. , Patnayak Devi P. , Goyal Sagar M.
Published: May 2011

Acute respiratory tract infections are leading causes of morbidity in poultry farms throughout the world. Avian pneumovirus (APV), avian influenza virus (AIV), and Newcastle disease virus (NDV) have been recognized as the most important pathogens of both chicken and turkeys. Single-virus reverse transcription–polymerase chain reaction (sRT-PCR) assays are used extensively to detect these viruses in clinical samples. This study reports the development and evaluation of a single-tube multiplex RT-PCR (mRT-PCR) assay for simultaneous and specific detection of APV, AIV, and NDV. Specific primers for each virus were selected that amplified products of predicted sizes from each virus in the mRT-PCR as well as in the sRT-PCR assays (438, 218, and 532 bp for APV, AIV, and NDV, respectively). The sensitivity and specificity of mRT-PCR assay were compared with those of the sRT-PCR. The mRT-PCR assay was as sensitive as the sRT-PCR assays because virus detection limits were similar in both assays. The detection limits of mRT-PCR assay were 100.5 tissue culture infective dose (50%) (TCID50)/ml, 101.2 TCID50/ml, and 100.7 TCID50/ml for APV, AIV, and NDV, respectively. Overall, there was an excellent correlation between mRT-PCR and sRT-PCR assays. No product amplification was obtained with nucleic acid from infectious bronchitis virus and reovirus using these primer sets. In summary, mRT-PCR assay holds potential to be an economical and rapid diagnostic method for the simultaneous detection of 3 avian respiratory viruses in chickens and turkeys.
Granulomatous Lymphadenitis and Splenitis Associated with Monocillium Indicum Infection in a Dog
Authors: Mackie John T. , Padhye Arvind A. , Sutherland Reginald J. , Lamb Warwick A. , Davis Stephen
Published: May 2011

This report describes severe generalized granulomatous lymphadenitis and splenitis in a 5-year-old, spayed female, Rottweiler dog with anorexia and diarrhea. There was replacement or effacement of much of the parenchyma of the lymph nodes and spleen by sheets of macrophages, multinucleated giant cells, and myriad nonpigmented fungal organisms, most of which appeared to be intracellular. These organisms were very pleomorphic, including large chlamydospore-like cells, small round yeast-like cells, and septate hyphae. A fungus identified as Monocillium indicum was isolated from lymph node tissue. To the authors' knowledge, this is the first report of infection with Monocillium in either humans or other animals.
The Quality Assurance of Proficiency Testing Programs for Animal Disease Diagnostic Laboratories
Authors: Wiegers Ann L.
Published: May 2011

Laboratory data credibility has 3 major components: 1) valid methods, 2) proficiency testing (PT) to verify that the analyst can conduct the method and to compare results of other laboratories using the same method, and 3) third-party accreditation to verify that the laboratory is competent to conduct testing and that the method validation has been done within the environment and requirements of an effective quality-management system. Participation in external PT programs by a laboratory is strongly recommended in International Organization for Standardization/International Electrotechnical Commission International Standard 17025. Most laboratory accreditation bodies using this standard require that laboratories participate in such programs to be accredited. Internal PT is also recommended for each analyst. Benchmarking, or comparison between laboratories using PT or reference materials, is also recommended as part of the validation and evaluation of test methods. These requirements emphasize the need for proficiency test providers to demonstrate their competence. Requirements for competence are documented in national and international standards and guidelines, and accreditation is available for providers. This article discusses the activities and the components that are necessary and recommended for PT projects and programs for animal disease diagnostic testing. These are based on the requirements of the national and international standards, which address this subject, and on the experience of the author. The accreditation of external PT programs is also discussed. Organizations that accredit PT providers or that provide PT programs are listed. Existing references, guidelines, and standards that are relevant to PT in veterinary diagnostic laboratories are discussed.
Influence of Age, Sex, and Production Class on Liver Zinc Concentration in Calves
Authors: Puschner Birgit , Choi Young-Ku , Tegzes John H. , Thurmond Mark C.
Published: May 2011

Determination of zinc concentrations in the liver of calves and young stock is commonly requested by practitioners and nutritionists to assess whether they receive an appropriate amount of zinc in their diet. However, interpretation of liver zinc concentrations is currently based on information reported for adult cattle for which the health status was unknown and irrespective of production class, sex, and age. A retrospective study of necropsy reports was undertaken to assess the relationships between liver zinc concentrations and age, sex, and production class for calves that did not have a history compatible with zinc toxicosis or zinc deficiency. Results of a generalized least squares, polynomial regression analysis of 474 records found that zinc concentration was not affected by sex ( P = 0.29) or production class ( P = 0.50). Zinc concentration was significantly associated with linear ( P < 0.00001) and nonlinear (quadratic, P = 0.0039) functions of age ( r2 = 0.1503), where the concentration decreased from 93 mg/kg wet weight at 30 days of age to 57 mg/kg wet weight at 9 months of age, after which it began to increase. The age-specific 95% confidence limits of the mean concentration for a group of calves and the 95% prediction limits of a single concentration value for an individual animal estimated in this study suggest reconsideration of the recommended limits for liver zinc concentration in calves. As a consequence of the significant influence of age on liver zinc concentration of calves presumably not experiencing zinc toxicosis or deficiency, diagnosis of zinc imbalances based on liver zinc concentration needs to consider age as a diagnostic covariate.
Investigations into Shaking Mink Syndrome: An Encephalomyelitis of Unknown Cause in Farmed Mink (Mustela Vison) Kits in Scandinavia
Authors: Dolores Gavier-Widén , Caroline Bröjer , Hans Henrik Dietz , Englund Lena , Anne Sofie Hammer , Hedlund Kjell-Olof , Carl Hård af Segerstad , Nilsson Kristina , Nowotny Norbert , Puurula Vuokko , Peter Thorén , Uhlhorn Henrik , Herbert Weissenböck , Erik Ågren , Klingeborn Berndt
Published: May 2011

An apparently novel neurological disease clinically characterized by shaking, tremors, seizures, staggering gait, and ataxia was first observed in farmed mink kits in Denmark in 2000 and subsequently in Sweden, Denmark, and Finland in 2001, and again in Denmark in 2002. Lymphoplasmacytic encephalomyelitis was found in the affected kits. The lesions were most severe in the brainstem and cerebellum and consisted of neuronal degeneration and necrosis, neuronophagia, focal and diffuse gliosis, perivascular cuffs formed by lymphocytes, plasma cells and macrophages, and segmental loss of Purkinje cells. Testing was conducted to determine the cause of the disease, including general virological investigations (virus culture, negative-staining electron microscopy, immunoelectron microscopy, polymerase chain reaction for herpesviruses, adenoviruses, pestiviruses, and coronaviruses), tests for specific viral diseases (canine distemper, Borna disease, Louping ill, West Nile virus infection, tick-borne encephalitis, Aleutian disease), tests for protozoa ( Toxoplasma gondii, Neospora caninum, Encephalitozoon cuniculi), bacteria (general culture, listeria, Clamydophila psittaci), and intracerebral inoculation of neonatal mice. The results of all these investigations were negative. One group of 3 mink kits inoculated intracerebrally with brain homogenate of affected mink developed clinical signs and histological lesions similar to those observed in naturally infected mink. Based on the histopathological features, it is postulated that the disease is caused by a yet unidentified virus.
Effect of Feeding Green Onions (Allium Ascalonicum) to White Chinese Geese (Threskiornis Spinicollis)
Authors: Crespo Rocio , Chin R. P.
Published: May 2011

Sudden increase in mortality was observed in 2 different flocks of mature breeder geese fed green onions. At necropsy, birds had pale epicardium with random petechiation, sanguinous fluid accumulation in the pericardial sac, and mild swelling of the liver and spleen. Histologically, there was accumulation of hemosiderin in hepatocytes, Kupffer cells of the liver, macrophages, and renal tubules. There was also moderate to severe hepatic necrosis, vacuolation of hepatocytes, splenitis, and renal tubular nephrosis. To assess the effects of green onion ingestion, 2 feeding trials were carried out in 3 mature White Chinese geese. In the first trial, onions were thoroughly mixed with pellet maintenance ration. In the second trial, onions were offered in a separate trough from the pelleted diet. During the 21 days of experiments, the red blood cell count and hematocrit decreased, whereas the polychromasia and reticulocyte estimate increased. The blood changes were more marked in birds from the second feeding trial. Gross and histologic changes were similar in both trials. Mild swelling and severe darkening of the liver were the only significant findings at necropsy. Histologically, the liver looked similar to that seen from the field outbreak. The liver contained moderate amounts of hemosiderin in the hepatocytes and Kupffer cells, and had centrolobular necrosis and vacuolation of hepatocytes. This experimental study demonstrated that anemia and liver pathology could be caused by ingestion of onions. Furthermore, Heinz bodies are not a consistent finding in the blood of geese fed onions.
Equine Abortion and Premature Birth Associated with Cellulosimicrobium Cellulans Infection
Authors: Bolin David C. , Donahue James M. , Vickers Mary L. , Giles Ralph C. , Harrison Lenn , Jackson Carney , Poonacha K. B. , Roberts John F. , Sebastian Manu M. , Sells Steve F. , Tramontin Robert , Williams Neil M.
Published: May 2011

During the 2002 and 2003 foaling seasons, Cellulosimicrobium ( Cellumonas) cellulans (formerly Oerskovia xanthineolytica) was the principal microorganism isolated from fetal tissues or placentas from cases of equine abortion, premature birth, and term pregnancies. Significant pathologic findings included chronic placentitis and pyogranulomatous pneumonia. In addition, microscopic and macroscopic alterations in the allantochorion from 4 of 7 cases of placentitis were similar to those caused by Crossiella equi and other nocardioform bacteria. This report confirms a causative role of C. cellulans infection in equine abortion.
Vertebral Malformation, Syringomyelia, and Ventricular Septal Defect in a Dromedary Camel (Camelius Dromedarius)
Authors: Sakamoto Kaori , Kiupel Matti , Frank Nicholas , March Philip A.
Published: May 2011

An occipitoatlantoaxial malformation and ventricular septal defect (VSD) were diagnosed in a 36-hour-old female camel. Physical examination revealed a firm protrusion of the dorsal aspect of the atlas and axis, tilting of the head to the left, and a grade V/VI systolic murmur. Neurological examination revealed proprioceptive deficits and ataxia of all 4 limbs. Radiographic examination and necropsy demonstrated malformation, fusion of the atlas to the occiput and hypoplasia of the dens of the axis, and subluxation of the atlantoaxial joint. Dorsoventral laxity of the atlantoaxial joint was also present, with compression of the cervical spinal cord. A 1.5-cm-diameter VSD was observed also. Histopathologic examination of the cervical spinal cord revealed a cavity extending from the level of the first to fourth cervical segment, dorsal to the central canal, 5 cm long and 1–2 mm in diameter. The cells around the cavity were positive for glial fibrillary acidic protein and sporadically positive for vimentin. This cavitary structure was consistent with syringomyelia, which was lined by glial cells, surrounded by edematous white matter with Wallerian-like degeneration and with neuronal necrosis in the adjacent dorsal horns.
Concurrent Presence of Sarcocystis Neurona Sporocysts, Besnoitia Darlingi Tissue Cysts, and Sarcocystis Inghami Sarcocysts in Naturally Infected Opossums (Didelphis Virginiana)
Authors: Elsheikha H. M. , Fitzgerald S. D. , Rosenthal B. M. , Mansfield L. S.
Published: May 2011

Opossums ( Didelphis virginiana) are exposed to a wide range of coccidia through feeding on a variety of foods, including, but not limited to, carrion, insects, and nestling birds. Abundant D. virginiana populations in urban and suburban areas can be important reservoirs of parasitic infection because of their profuse and prolonged excretion of the sporocysts of several species of Sarcocystis, their omnivorous diet, and their relatively long life span. This report describes 2 adult female opossums found to be simultaneously infected with the tissue cysts of Besnoitia darlingi, sarcocysts of Sarcocystis inghami, as well as with the intestinal sporocysts of S. neurona. Cysts typical of B. darlingi based on gross, histological, and ultrastructural characteristics were disseminated throughout the visceral organs, musculature, ears, and skin. The S. neurona and B. darlingi infections were confirmed by comparative sequence analysis of polymerase chain reaction—amplified diagnostic genetic loci. Sarcocysts of S. inghami are also described. Such examples of multiple parasitic infections show that concurrent infections occur naturally. The propensity for species to coexist should be considered in the differential diagnosis of tissue cyst—forming coccidian protozoa and may have important epidemiological and evolutionary implications.
Fibrocartilaginous Embolic Myelopathy in a Calf
Authors: Landolfi Jennifer A. , Saunders Geoffrey K. , Swecker William S.
Published: May 2011

A 5-month-old Angus heifer with a history of acute hindlimb paresis that quickly progressed to lateral recumbency was necropsied. Gross lesions included a 6-cm segment of gray to brown discoloration and softening of the right ventrolateral spinal cord between T2 and T3. Microscopically, there was liquefactive necrosis of ventrolateral white and gray matter, and multiple intravascular emboli partially or completely occluded many intralesional and adjacent spinal and meningeal arteries and veins. Emboli were alcian blue positive, consistent with fibrocartilage of the nucleus pulposus of the intervertebral disk. No gross abnormalities were detected in the vertebrae or intervertebral disks. Fibrocartilaginous embolic myelopathy appears to be very rare in cattle; however, it should be considered in cases of acute, nonprogressive spinal cord dysfunction.
Real–Time Reverse Transcription–Polymerase Chain Reaction Assays for the Detection and Differentiation of North American Swine Influenza Viruses
Authors: Richt Jürgen A. , Lager Kelly M. , Clouser Deborah F. , Spackman Erica , Suarez David L. , Yoon Kyoung-Jin
Published: May 2011

Swine influenza is an acute respiratory disease of swine caused by type A influenza viruses. Before 1998, mainly “classical” H1N1 swine influenza viruses (SIVs) were isolated from swine in the United States. Since then, antigenically distinct reassortant H3 and H1 SIVs have been identified as causative agents of respiratory disease in pigs on US farms. Improvement in SIV diagnostics is needed in light of the recently observed rapid evolution of H1 and H3 SIVs and their zoonotic potential. To address this need, real-time reverse transcription–polymerase chain reaction (RT-PCR) assays for the detection of SIVs were developed. A highly sensitive matrix (M) gene–based RT-PCR assay that is able to detect both the H1 and H3 subtypes of SIVs, with a sensitivity per reaction of approximately 2 copies of in vitro–generated M-specific negative-sense RNA molecules and approximately 0.05 TCID50 in lung lavage of experimentally SIV-infected pigs, was established. This RT-PCR assay can be performed within a few hours and showed a sensitivity of 94% and a specificity of 85% when compared with virus isolation. In addition, H1-, H3-, N1-, and N2-specific primer and probe sets were designed for use in the differentiation of different SIV subtypes. The hemagglutinin (H)- and neuraminidase (N)-specific primer and probe sets were less sensitive than the M-specific assay, although they were found to be specific for their respective viral genes and able to distinguish between their respective SIV subtypes.
Distribution of Viral Antigen and Tissue Lesions in Persistent and Acute Infection with the Homologous Strain of Noncytopathic Bovine Viral Diarrhea Virus
Authors: Liebler-Tenorio Elisabeth M. , Ridpath Julia F. , Neill John D.
Published: May 2011

Viral distribution and lesions were compared between calves born with persistent infection (PI) and calves acutely infected with the same bovine viral diarrhea virus (BVDV) isolate. Two PI calves from 1 dairy herd were necropsied. The PI viruses from these calves were isolated, characterized by sequencing, and found to be identical. This virus strain, designated BVDV2-RS886, was characterized as a noncytopathic (ncp) type 2 BVDV. To establish acute infections, BVDV2-RS886 was used to inoculate clinically healthy, seronegative calves which were 3 weeks to 3 months old. Nine calves received 106–107 tissue culture infective dose of BVDV2-RS886 intranasally. Four additional age-matched animals served as noninfected controls. Infected calves were necropsied at 3, 6, 9, or 13 days postinoculation (dpi). Viral antigen was detected by immunohistochemistry in frozen sections, and lesions were evaluated in hematoxylin eosin–stained paraplast sections. In the PI calves, a wide distribution of viral antigen was found in all tissues and was not associated with lesions. In the acutely infected calves, viral antigen was widespread in lymphoid tissues at 6 dpi but had been mostly eliminated at 9 and 13 dpi. Depletion of lymphoid tissues was seen at 6, 9, and 13 dpi and repopulation at 9 and 13 dpi. In 1 of the calves at 13 dpi, severe arteritis was present in lymph nodes and myocardium. This comparison shows that an ncp BVDV strain that causes no lesions in PI animals is able to induce marked depletion of lymphoid tissues in calves with acute infection. Therefore, the failure to eliminate PI cattle from a herd causes problems not only in pregnant cattle but may also affect other age groups.
Mycoplasma Otitis in California Calves
Authors: Lamm Catherine G. , Munson Linda , Thurmond Mark C. , Barr Bradd C. , George Lisle W.
Published: May 2011

A retrospective study of Mycoplasma otitis in California calves submitted for necropsy between 1993 and 2002 was conducted to characterize the demographic features of the disease and the pathologic findings associated with infection. Sixty-one confirmed cases of Mycoplasma otitis were identified among 20,525 necropsied cattle. All affected animals were calves, ranging in age from 2 weeks to 4 months and with a median age of 1.5 months. Ninety-two percent of the cases were dairy breeds. A higher percent of necropsied calves with Mycoplasma otitis were males (0.45%) than females (0.23%). The proportion of cases that had Mycoplasma otitis increased from 1993 to 2002, and there was a significant ( P < 0.05) seasonal distribution, with the highest proportion in the spring and the lowest in the summer months. Infections involved both the middle and inner ear and were characterized by a suppurative inflammatory response with extensive bony involvement. Three species of Mycoplasma were isolated from the ears: M. bovis, M. bovirhinis, and M. alkalescens. Concurrent pneumonia occurred in 47 cases (77%), and Mycoplasma was isolated from the lungs of 30 of those cases. The increasing proportion of Mycoplasma otitis cases in the past 10 years emphasizes the importance of identifying risk factors that could be modified to lower the incidence of this disease in calves.
Immunoglobulin G2 Deficiency with Transient Hypogammaglobulinemia and Chronic Respiratory Disease in a 6-Month-Old Holstein Heifer
Authors: Francoz David , Lapointe Jean-Martin , Wellemans Vincent , Desrochers André , Caswell Jeff L. , Stott Jeff L. , Dubreuil Pascal
Published: May 2011

A 6-month-old Holstein heifer that was nonresponsive to medical treatment was evaluated for chronic respiratory disease. Complete blood count and serum chemistry revealed neutrophilic leukocytosis and low globulin levels. Assays for bovine leukemia virus, bovine virus diarrhea, and bovine leukocyte adhesion deficiency were negative. Serum globulin subclass assays revealed transient low concentrations of immunoglobulin (Ig) G1 and IgA, persistent low IgG2, and subnormal IgM. Vaccination with 2 doses of multiple, inactived viruses induced seroconversion for most viruses. Flow cytometric analysis of blood lymphocyte subpopulation demonstrated an increase in CD5+ B-cells. Blood lymphocyte proliferation and neutrophil function tests were normal. Results of immunologic assays indicated IgG2 deficiency with transient hypogammaglobulinemia.
Use of Conventional and Real-Time Polymerase Chain Reaction for Confirmation of Mycobacterium Avium Subsp. Paratuberculosis in a Broth-Based Culture System ESP II
Authors: Kim Sung G. , Kim Eun H. , Lafferty Caroline J. , Miller Loretta J. , Koo Hye J. , Stehman Susan M. , Shin Sang J.
Published: May 2011

The ESP II Culture System (ESP II), a broth-based culture system, has been modified and optimized for culturing Mycobacterium avium subsp. paratuberculosis ( M. paratuberculosis) in animal feces since 2000. Conventional and real-time polymerase chain reaction (PCR) assays based on the IS 900 sequence were performed as confirmatory tests for M. paratuberculosis in ESP II liquid culture medium. There were no differences between test results of conventional and real-time PCR assays. During the 5-week incubation period, if acid-fast bacilli (AFB) were detected in ESP culture–positive samples, IS 900 PCR assays were performed to confirm whether those AFB were M. paratuberculosis. At the end of the 5-week incubation, AF staining was performed on all ESP II–negative cultures to screen any false-negative cultures; IS 900 PCR assays were performed on AFB-positive cultures. During a period of 1 year, of a total of 18,499 ESP II cultures, 2,814 (15.2%) PCR confirmation assays were performed. Of those, 2,259 (80%) were both ESP and PCR positive; 104 (4%) were ESP positive and PCR negative; 423 (15%) were ESP negative and PCR positive; 28 (1%) were both ESP and PCR negative. The AF-staining step after the 5-week incubation produced 423 (15%) more PCR-positive cultures. Of a total of 2,814 AFB-positive cultures, 132 (5%) were not confirmed as M. paratuberculosis. Further studies are needed for speciation of non– M. paratuberculosis isolates.
Isolation of an Adenovirus and an Adeno-Associated Virus from Goat Kids with Enteritis
Authors: Olson Erik J. , Haskell Scott R. R. , Frank Rodney K. , Lehmkuhl Howard D. , Hobbs Lea Ann , Warg Janet V. , Landgraf John G. , Wünschmann Arno
Published: May 2011

A dairy goat operation in Minnesota experienced a sudden, markedly increased mortality among its neonatal goats. Approximately 60 of 130 kids (46%) died. The animals had diarrhea and dyspnea of 1–2 days duration before death. Necropsy of 4 goat kids revealed marked, acute, catarrhal enteritis and fibrinous pleuropneumonia. Mannheimia haemolytica was isolated from the lungs. Basophilic inclusion bodies filling the entire nucleus were present in enterocytes of the ileum of 3 goats. Adenoviral particles were detected in the feces by electron microscopy and adenovirus was subsequently isolated from the intestinal content together with a parvo-like virus (dependovirus). Morphology, physicochemical characteristics, and neutralization tests indicated that the adenovirus resembled ovine adenovirus-2 (OAdV-2). However, the PstI restriction endonuclease pattern produced by the goat adenovirus was distinct from that of OAdV-2. This is the first report of enteritis in goats with an adenovirus antigenically related to OAdV-2 and with a parvo-like dependovirus.
Eye Infections due to Listeria Monocytogenes in Three Cows and One Horse
Authors: Evans Katie , Smith Mary , McDonough Pat , Wiedmann Martin
Published: May 2011

A retrospective study was conducted to determine case histories, microbiological characteristics, and molecular subtypes associated with Listeria monocytogenes infections of the eye in large animals. For selected cases, environmental L. monocytogenes contamination patterns on case farms were also evaluated to probe for potential sources and spread of listerial eye infections. Records of 170 L. monocytogenes isolates from animal infections were reviewed to determine the fraction of isolates associated with eye infections (conjunctivitis, keratitis, and uveitis) of animals and to gather information on the clinical history of these cases. Overall, 4 of 170 Listeria monocytogenes isolates were associated with eye infections; 3 of these had occurred in cows and 1 in a horse. Molecular subtyping (by EcoRI ribotying) showed that 4 different L. monocytogenes subtypes were isolated from these 4 cases; the same ribotypes had previously been found among invasive animal listeriosis infections. Although a variety of L. monocytogenes subtypes were isolated from environmental sources, on 1 farm, the same ribotype associated with the eye infection was also isolated from a fecal sample of a healthy animal and from a soil sample. The data reported in this study further suggest that L. monocytogenes can be a cause of eye infections in several animal species. Listerial eye infections do not seem to require specific pathogen-related virulence characteristics but rather seem to be a function of environmental or host factors, such as direct exposure of the eyes of susceptible animals to high numbers of the pathogen. Although listerial eye infections are rarely diagnosed because of its ubiquitous nature, L. monocytogenes may have to be considered more commonly as a causative agent of eye infections in ruminants and horses.
Rapid and Simple Mutation Screening of GM1 Gangliosidosis in Shiba Dogs by Direct Amplification of Deoxyribonucleic Acid from Various Forms of Canine Whole-Blood Specimens
Authors: Yamato Osamu , Jo Eun-Og , Shoda Toru , Yamasaki Masahiro , Maede Yoshimitsu
Published: May 2011

This report describes a rapid and simple method for mutation screening of GM1 gangliosidosis in Shiba dogs by direct amplification of DNA from canine whole-blood specimens using a novel polymerase chain reaction (PCR) reagent cocktail, which can eliminate the DNA extraction process and amplify the genomic DNA directly from human or murine whole blood. The strategy of this mutation screening is based on the identification of a nucleotide deletion by restriction enzyme analysis, coupled with the direct PCR amplification. The target sequence of the canine β-galactosidase gene could be amplified directly from various forms of canine whole-blood specimens, including anticoagulated blood, blood stored frozen for 1 year, dried blood held in filter paper for 1 year at room temperature, and dry powder of blood stripped from Giemsa-stained blood films, which had been prepared 10 years earlier, resulting in the determination of genotypes in all the specimens. This method simplified the molecular diagnosis and carrier screening of GM1 gangliosidosis in Shiba dogs, making it simple to examine specimens from the large, widely distributed population of these dogs.
Utility of High-Performance Liquid Chromatography in the Characterization of Iatrogenic Sodium Diatrizoate–Induced Seizures in a Horse
Authors: Fitzgerald Scott D. , Braselton W. Emmett
Published: May 2011

Diagnostic evaluation of a mildly ataxic young gelding Quarterhorse included a contrast myelogram. Unfortunately, sodium diatrizoate was inappropriately administered by intrathecal injection resulting in severe unrelenting seizure activity and necessitated the destruction of the animal. The contrast agent was identified by analyzing cerebrospinal fluid by high-performance liquid chromatography and confirmed with probe mass spectrometric analysis. This contrast agent is highly ionic, has high osmolarity, and also blocks neurotransmission at specific receptors; for these reasons it is contraindicated for use in myelography. This is the first report of intrathecal administration of this agent in a horse, although there has been a previous report of similar untoward reactions in a dog, and numerous cases have been reported in humans.
Abortion and Ulcerative Posthitis Associated with Caprine Herpesvirus–1 Infection in Goats in California
Authors: Uzal Francisco A. , Woods Leslie , Stillian Michelle , Nordhausen Robert , Read Deryck H. , Van Kampen Hanna , Odani Jenee , Hietala Sharon , Hurley E. J. , Vickers Mary L. , Gard Sharon M.
Published: May 2011

Three outbreaks of late-gestation abortions in does and ulcerative posthitis in bucks, associated with caprine herpes virus–1 (CHV-1), in California are described. In herd A, 10 of 17 does aborted in a 7-day period, whereas in herd B, 4 of 130 does aborted in a 45-day period and in herd C, 100 of 300 does aborted in a 3-week period. Most fetuses had multifocal pinpoint depressed foci with a zone of hyperemia on external and cut surfaces of the kidneys, liver, lungs, and adrenal glands. Histologically, scattered multifocal areas of necrosis with mild neutrophilic infiltrate were observed in kidneys, brain, liver, adrenal glands, and lungs of most fetuses of the 3 herds. Large amphophilic intranuclear inclusion bodies, which displaced the chromatin, were observed in cells within and around the necrotic foci in kidneys and adrenal glands. Particles 85–113 nm in size with morphology compatible with herpes virus were observed in the nuclei of these cells when examined by electron microscopy. Irregular, shallow, red ulcers were observed in the prepuce of 1 buck from herd C. Prepuce biopsies from this animal had necrosis of the superficial mucosal epithelium and severe submucosal lymphoplasmocytic infiltrates. Large intranuclear amphophilic inclusion bodies were observed in most cells of the stratum spinosum of the preputial epithelium, but no viral particles were observed in these cells. Caprine herpes virus–1 was isolated from tissue pools of fetuses from the 3 herds but not from prepuce biopsies. Positive results were obtained when tissues of a fetus from herd C were processed by a polymerase chain reaction technique to amplify the amino terminus of the glycoprotein C gene of CHV-1. Sera from aborted does from herds B and C and from the 3 bucks from herd C had high antibody titers to CHV-1. The results presented here support the hypothesis that the male goat is involved in the transmission of CHV-1. However, other forms of transmission cannot be ruled out.
The Effect of Pooling Sera on the Detection of Avian Pneumovirus Antibodies using an Enzyme-Linked Immunosorbent Assay Test
Authors: Maherchandani Sunil , Muñoz-Zanzi Claudia A. , Patnayak Devi P. , Malik Yashpal S. , Goyal Sagar M.
Published: May 2011

Pooling of samples is a cost-effective approach to estimate disease prevalence and to identify infected individuals. The objective of this study was to evaluate the use of serum pools for the detection of avian pneumovirus infection in turkey flocks by enzyme-linked immunosorbent assay, so that a minimum number of tests can be performed without compromising the sensitivity and specificity of the test. A total of 900 field samples were tested; 20 samples from each of 45 flocks. All samples were tested individually followed by pool testing in groups of 3, 4, 5, and 7 samples each. The number of positive pools for a given pool size was positively associated with the number of positive samples. In a separate experiment, the effect of dilution was examined by pooling 1 positive sample with different numbers of negative samples to form pools of sizes 2–7. These laboratory results were analyzed and integrated into a simulation model aimed at evaluating cost-efficient testing procedures. The probability of detecting an infected flock depended on prevalence of infection, size of serum pool, and the cutoff value used for optical density difference. At a theoretical prevalence of 20%, the probability of detecting an infected flock was 0.93 and 0.86 for a pool of 2 and 7, respectively. The probability of detecting positive flocks increased with increased prevalence and decreased cutoff. Pooling of samples represented a significant reduction in the cost of testing, suggesting that pooling is more advantageous and cost effective than testing individual samples.
Detection of Mycobacterium Avium Subsp. Paratuberculosis in Bovine Fecal Samples: Comparison of Three Polymerase Chain Reaction—Based Diagnostic Tests with a Conventional Culture Method
Authors: Taddei Simone , Robbi Chiara , Cesena Camilla , Rossi Ilenia , Schiano Emiliana , Arrigoni Norma , Vicenzoni Gaddo , Cavirani Sandro
Published: May 2011

Three commercially available assays, designed to specifically detect the presence of Mycobacterium avium subsp. paratuberculosis (MAP) in fecal samples by IS900-PCR, were compared with a conventional culture method. Fecal samples from 100 dairy cows were tested. Fifty-four (67.5%) of 80 culture-positive samples were positive for an assay that detects MAP DNA by dot spot hybridization of polymerase chain reaction products (kit A), 48 (60%) were positive by an assay using ethidium bromide staining for agar gel visualization of amplification products (kit B), and 49 (61.3%) were positive by an assay in which amplified products are detected by a colorimetric detection system (kit C). Relative sensitivity of all tests increased in proportion to the presence of MAP in fecal samples. Specificity was 100% based on results from 20 culture-negative samples from an MAP-free herd.
A Retrospective Study of Mortality in Pennsylvania Captive White-Tailed Deer (Odocoileus Virginianus): 2000–2003
Authors: Hattel Arthur L. , Shaw Daniel P. , Love Brenda C. , Wagner Donald C. , Drake Thomas R. , Brooks Jason W.
Published: May 2011

The postmortem records of 160 white-tailed deer ( Odocoileus virginianus) submitted for necropsy examination from 59 separate Pennsylvania captive deer farms over a 3.5-year period were reviewed to determine the primary cause of death of each animal. The most common causes of death were bronchopneumonia (39 cases), enterocolitis (30 cases), malnutrition (13 cases), and trauma (11 cases). Other causes of mortality included severe gastrointestinal parasitism (6 cases), cellulitis with septicemia (5 cases), degenerative myopathy (4 cases), ruminal acidosis (4 cases), and nephritis (4 cases). The cause of death was undetermined in 13 of the 160 animals. Arcanobacterium pyogenes (19 cases), Fusobacterium necrophorum (10 cases), Escherichia coli (7 cases), and Mannheimia haemolytica (4 cases) were the most commonly isolated bacteria from the pneumonic lungs. Bacterial agents associated with enterocolitis included Clostridium perfringens (15 cases), E. coli (12 cases), and Mycobacterium avium subsp. paratuberculosis (2 cases). The majority (52.2%) of the death loss in white-tailed deer of known ages occurred in animals 1 year of age or less, with 46.2% of the bronchopneumonia cases and 50.0% of the enterocolitis cases occurring during this time period. Cases of degenerative myopathy, myocardial degeneration, hepatic necrosis, meningoencephalitis, peritonitis, and urolithiasis considered severe enough to be the primary cause of death appeared early in life, affecting deer 6 months of age or less in all cases. In conclusion, bronchopneumonia, enterocolitis, malnutrition, and trauma were considered the most common causes of death in confined white-tailed deer in this study.
A Retrospective Study of Visceral and Nonvisceral Hemangiosarcoma and Hemangiomas in Domestic Animals
Authors: Schultheiss Patricia C.
Published: May 2011

Cases of hemangiosarcoma submitted to the Colorado State University Veterinary Diagnostic Laboratory during a 6-year period were reviewed. Visceral hemangiosarcomas represent less than 2% of canine specimens submitted for histologic examination and nonvisceral hemangiosarcoma less than 1%. Most nonvisceral hemangiosarcomas of dogs occur in skin. Hemangiosarcomas are less common in cats and usually occur in skin. They are also rare in other animal species. Animals with nonvisceral hemangiosarcomas are usually mature; dogs and cats average 10 years of age. The tumors develop in many different locations, and there is no sex predilection. A wide variety of dog breeds are affected, but Italian greyhounds, greyhounds, and whippets are overrepresented. Clinical outcomes of 76 cases of nonvisceral hemangiosarcomas in dogs and cats were obtained from submitting veterinarians. Completeness of excision of a tumor is the most important factor that can be used in predicting clinical outcome for an affected dog or cat. In all cases in which the animals were clinically normal for at least 1 year after surgical removal of a nonvisceral hemangiosarcoma, the margins were reported to be free of neoplastic cells. Degree of differentiation, mitotic rate, size of tumor, and presence or absence of epidermal ulceration, mast cells, or solar elastosis did not correlate with clinical outcome.
E-Cadherin Immunoreactivity in Canine Mammary Tumors
Authors: Sarli Giuseppe , Preziosi Rosario , Tolla Louis De , Brunetti Barbara , Benazzi Cinzia
Published: May 2011

The reduction or loss of E-cadherin (E-cad), a calcium-dependent epithelial cell adhesion molecule, has been associated with tumor dedifferentiation and invasiveness. The immunohistochemical pattern of E-cad expression was evaluated in formalin-fixed and paraffin-embedded sections of 6 normal mammary glands, 3 dysplasias, 12 benign tumors (8 benign mixed tumors, 4 adenomas), and 60 malignant tumors (12 stage 0, 29 stage I, 19 stage II) of the canine mammary gland. E-cadherin expression was classified as membranous, when on cell–cell boundaries, or as cytoplasmic, when in the form of a diffuse cytoplasmic staining. In addition, the percentage of E-cad–positive epithelial neoplastic cells was graded by a semiquantitative method, categorizing cases into a reduced (or -) type group, when showing less than 25% positivity, a reduced (or +/-) type group, when showing 25–75% positivity, and a preserved (or +) type group, when more than 75% positive cells were present. In the normal mammary gland, E-cad expression was evident in epithelial luminal cells. A stronger positivity was revealed in ductular than in alveolar luminal cells. The myoepithelial cells showed inconsistent, weak cytoplasmic positivity in the normal gland as well as in mammary tumors. In normal glands and benign and malignant noninvasive tumors, E-cad expression was mainly membranous and preserved in most of the epithelial cells. In stage I tumors, both membranous (38%) and cytoplasmic (62%) positivity were well represented, as well as preserved type (55%) and reduced type (45%) tumors. All stage II malignant tumors showed the highest frequency of cytoplasmic positivity (79%) and reduced type (62%) tumors.
Competitive Enzyme-Linked Immunosorbent Assay for the Detection of the Antibodies Specific to Akabane Virus
Authors: Tsuda Tomoyuki , Yoshida Kazuo , Yanase Tohru , Ohashi Seiichi , Yamakawa Makoto
Published: May 2011

A competitive enzyme-linked immunosorbent assay (C-ELISA) using neutralizing monoclonal antibodies (MAbs) against Akabane virus (AKAV) was developed to detect antibodies to AKAV in cattle sera. The performance of the test using 7 different competitor MAbs was evaluated in sequential serum samples and sera from cattle infected with various bovine arboviruses. The dynamics of the antibody response expressed by percentage of inhibition (PI) in C-ELISA coincided with those of neutralizing antibody titers in sequential serum samples from 2 cattle experimentally infected with AKAV. The value of PI in C-ELISA for convalescent sera from cattle infected with arboviruses correlated with the neutralizing antibody titer to AKAV but was unaffected by the antibodies to other arboviruses. In the validation experiment of C-ELISA using 286 bovine sera previously examined for the AKAV antibody by serum neutralization (SN) test, the relative specificity of C-ELISA was more than 98%, whereas the relative sensitivities of individual MAbs ranged from 49% to 82.2%. Overall agreement between C-ELISA and the SN test varied from 72% to 90% depending on the MAb. These results suggest that the C-ELISA is acceptable as a rapid and specific method for detecting antibodies to AKAV and is a potential alternative to the SN test.
Polymerase Chain Reaction Analysis of Eae Gene Subtypes Present in Attaching and Effacing Escherichia Coli Isolated from Pigs with Diarrhea
Authors: Ha Seung-Kwon , Hong Keumsuk , Choi Changsun , Jung Kwonil , Ha Yoonchul , Kim Junghyun , Kim Sung-Hoon , Yoon ByoungSu , Chae Chanhee
Published: May 2011

In this study the subtype of eae gene was determined by polymerase chain reaction for a total of 59 attaching and effacing Escherichia coli isolated from preweaned (38 isolates) and postweaned (21 isolates) pigs. The eaeβ gene detected in 19 E. coli from preweaned pigs and 10 E. coli from postweaned pigs was found to be the most common subtype, followed by eaeγ, eaeε, and eaeζ genes. Subtypes were not determined for 7 E. coli isolates. No other subtype of the eae gene was detected in eae+ E. coli evaluated in this study.
An Inexpensive Sedimentation Chamber for the Preparation of Cytologic Specimens of Cerebrospinal Fluid
Authors: Garma-Aviña Armando
Published: May 2011

An inexpensive sedimentation chamber to obtain cytologic specimens of cerebrospinal fluid (CSF) is described. The device, which has a total cost of about $5.00 can be built in few minutes. The device permits the cytologic study of specimens of CSF in clinics, where because of economic constraints, a cytocentrifuge is not available. The device permits the study of the CSF cells on either air-dried or wet smears even under field conditions, and the results obtained are consistent. Also, the device permits to retrieve the cell-free fluid for its use in chemical or immunologic procedures.
Experimental Transmission of Sheep Scrapie by Intracerebral and Oral Routes to Genetically Susceptible Suffolk Sheep in the United States
Authors: Hamir Amir N. , Kunkle Robert A. , Richt Juergen A. , Miller Janice M. , Cutlip Randall C. , Jenny Allen L.
Published: May 2011

Scrapie is a naturally occurring fatal neurodegenerative disease of sheep and goats. Susceptibility to the disease is partly dependent on the genetic makeup of the host. This study documents clinicopathological findings and the distribution of abnormal prion proteins (PrPres) by immunohistochemical and Western blot techniques, in tissues of genetically susceptible sheep inoculated with US sheep scrapie agents. Four-month-old Suffolk lambs (QQ or HQ at codon 171) were inoculated (5 intracerebrally and 19 orally) with an inoculum (#13–7) consisting of a pool of scrapie-affected sheep brains. Intracerebrally inoculated animals were euthanized when advanced clinical signs of scrapie were observed. Orally inoculated animals were euthanized at predetermined time points (4, 9, 12, 15, and 21 months postinoculation [PI]) and thereafter when the animals had terminal signs of disease. All intracerebrally inoculated animals exhibited clinical signs of scrapie and were euthanized between 13 and 24 months PI. Spongiform lesions in the brains and PrPres deposits in central nervous system and lymphoid tissues were present in these sheep. In orally inoculated sheep, clinical signs of scrapie were seen between 27 and 43 months PI in 5/9 animals. The earliest detectable PrPres was observed in brainstem and lymphoid tissues of a clinically normal, orally inoculated sheep at 15 months PI. Three of the 4 clinically normal sheep were positive at 15, 20, and 49 months PI by PrPres immunohistochemistry.
Myositis, Lameness, and Recumbency after use of Water-in-Oil Adjuvanted Vaccines in Near-Term Beef Cattle
Authors: O'Toole D. , Steadman L. , Raisbeck M. , Torpy R.
Published: May 2011

A producer administered 2 US Department of Agriculture-licensed adjuvanted veterinary vaccines (inactivated bovine rotavirus-coronavirus vaccine; Clostridium perfringens type C- Escherichia coli bacterin-toxoid) into muscles of the left and right hips of 469 pregnant beef cows. Within 24 hours, 5 cattle were recumbent, and another 2 had non-weight bearing pelvic limb lameness (1.5% affected; 7/469). During the next 10 days, 50% of the herd developed firm swellings up to 24 cm in vaccination sites in muscles of the hip. Histological samples revealed granulomatous myositis with intralesional oil. Lesions resolved slowly during the next 6 months. Six cattle were injected experimentally with the vaccines. None became lame, but all developed foreign body granulomatous myositis similar to those in the affected herd. The maximum diameter of experimentally induced lesions in muscle at necropsy 60 days after injection with the recommended dose of the bacterin-toxoid vaccines was 12 cm. Histological examination revealed pyogranulomatous myositis, fibrosis, and myonecrosis. The inactivated viral vaccine induced milder granulomatous myositis with intralesional lipid and scant fibrosis. Acute transient lameness on the ranch was attributed to use of 2 irritating biological vaccines in the hip muscles of cows that were close to parturition.