bluuebunny/crossref_metadata_2025 huggingface dataset:
Mycoplasma Bovis Real-Time Polymerase Chain Reaction Assay Validation and Diagnostic Performance
Authors: Clothier Kristin A. , Jordan Dianna M. , Thompson Curtis J. , Kinyon Joann M. , Frana Timothy S. , Strait Erin L.
Published: May 2011

Mycoplasma bovis is an important bacterial pathogen in cattle, producing a variety of clinical diseases. The organism, which requires specialized culture conditions and extended incubation times to isolate and identify, is frequently associated with concurrent infection with other pathogens which can potentially be more easily identified. Real-time polymerase chain reaction (real-time PCR) is a valuable diagnostic technique that can rapidly identify infectious agents in clinical specimens. A real-time PCR assay was designed based on the uvrC gene to identify M. bovis in diagnostic samples. Using culture as the gold standard test, the assay performed well in a variety of diagnostic matrices. Initial validation testing was conducted on 122 milk samples (sensitivity: 88.9% [95% confidence interval (CI): 68.4-100%], specificity: 100%); 154 lung tissues (sensitivity: 89.0% [95% CI: 83.1-94.9%], specificity: 97.8% [95% CI: 93.5-100%]); 70 joint tissue/fluid specimens (sensitivity: 92.3% [95% CI: 82.1-100%], specificity: 95.5% [95% CI: 89.3-100%]); and 26 nasal swabs (sensitivity: 75.0% [95% CI: 45.0-100%], specificity: 83.3% [95% CI: 66.1-100%]). Low numbers of other sample matrices showed good agreement between results of culture and PCR. A review of clinical cases from 2009 revealed that, in general, PCR was used much more frequently than culture and provided useful diagnostic information in conjunction with clinical signs, signalment, and gross and histopathologic lesions. Diagnostic performance of the real-time PCR assay developed as a testing method indicates that it is a rapid, accurate assay that is adaptable to a variety of PCR platforms and can provide reliable results on an array of clinical samples.
Proventricular Dilatation Disease Associated with Avian Bornavirus in a Scarlet Macaw (Ara Macao)
Authors: Keller Dominique L. , Honkavuori Kirsi S. , Briese Thomas , Lipkin W. Ian , Muthuswamy Anantharaman , Steinberg Howard , Sladky Kurt K.
Published: May 2011

A case of proventricular dilatation disease is described in a scarlet macaw( Ara macao) from clinical presentation to diagnosis with molecular methods. The initial clinical signs were depression progressing to head pressing over several days. A leukocytosis with toxic heterophil changes, hypoalbuminemia, and increased serum activity of aspartate aminotransferase and creatine kinase were present. Lead and zinc assays were within reference ranges, and results from Chlamydophila and polyomavirus testing were negative. Contrast-enhanced fluoroscopy revealed normal gastrointestinal transit times and motility as well as the presence of 2 small metallic foreign bodies in the ventriculus. The macaw was treated with antimicrobials, analgesics, vitamins E and B complex, force-feeding, and fluid administration with little improvement. Euthanasia was elected, and histologic examination of brain tissue revealed a perivascular lymphoplasmacytic infiltration, while the lungs had evidence of a fungal pneumonia. Tissue samples from the brain and proventriculus tested positive for the presence of Avian bornavirus genotype 2, while serology confirmed Avian bornavirus infection.
Chronic Myositis in an Australian Alpaca (Llama Pacos) Associated with Sarcocystis Spp
Authors: Gabor Melinda , Gabor Les J. , Srivastava Mukesh , Booth Mark , Reece Rod
Published: May 2011

An alpaca ( Llama pacos), born and raised in Australia, was presented with multiple subcutaneous abscesses. Histological findings indicated a severe necrotizing and histiocytic myositis and cellulitis associated with central caseation and multiple sarcocysts. Ultrastructural examination supported the diagnosis; however, cyst wall characteristics were not consistent with the 2 known species found in alpacas. While seroconversion in camelids is reported to be near ubiquitous, myositis is rare, and this is the first case reported outside of the Americas.
Retroperitoneal Pulmonary Choristoma in a Newborn Calf
Authors: Bassi Patrizia , Gentile Arcangelo , Militerno Gianfranco
Published: May 2011

A newborn male Romagnola calf, who had died a few hours after birth, was submitted for postmortem examination. Necropsy revealed a 23 cm × 22 cm × 5 cm, pale pink, lobulated, elastic, partially fluctuant mass that protruded from the dorsal retroperitoneal space into the abdominal cavity, extending from the diaphragm to the left kidney. The mass consisted of mature pulmonary tissue and was consistent with a pulmonary choristoma. The gross and microscopic appearance of this rare tumor-like congenital lesion and the possible pathogenesis are discussed.
Intrapericardial Ectopic Thyroid Carcinoma in a Cat
Authors: Knowles Susan , Uhl Elizabeth W. , Blas-Machado Uriel , Butler Abigail M.
Published: May 2011

A 7-year-old spayed female domestic shorthair feline presented with tachycardia and was later euthanized due to a declining condition. On gross examination, the thoracic cavity contained an expansile, multiloculated mass that displaced the lungs dorsocaudally. The mass, within the pericardial sac, compressed adjacent myocardium. Cut surface revealed variably sized, fluid-filled spaces with multiple foci of hemorrhage and necrosis. Histologically, the mass was composed of solid foci of polygonal cells admixed with colloid-containing follicles. Immunohistochemical staining for thyroglobulin was positive, and staining for calcitonin was negative. Grossly, thyroid glands were normal, and serum thyroxine was within reference intervals.
Epidemic and Economic Impacts of Delayed Detection of Foot-And-Mouth Disease: A Case Study of a Simulated Outbreak in California
Authors: Carpenter Tim E. , O'Brien Joshua M. , Hagerman Amy D. , McCarl Bruce A.
Published: May 2011

The epidemic and economic impacts of Foot-and-mouth disease virus (FMDV) spread and control were examined by using epidemic simulation and economic (epinomic) optimization models. The simulated index herd was a ≥2,000 cow dairy located in California. Simulated disease spread was limited to California; however, economic impact was assessed throughout the United States and included international trade effects. Five index case detection delays were examined, which ranged from 7 to 22 days. The simulated median number of infected premises (IP) ranged from approximately 15 to 745, increasing as the detection delay increased from 7 to 22 days. Similarly, the median number of herds under quarantine increased from approximately 680 to 6,200, whereas animals slaughtered went from approximately 8,700 to 260,400 for detection delays of 7–22 days, respectively. The median economic impact of an FMD outbreak in California was estimated to result in national agriculture welfare losses of $2.3–$69.0 billion as detection delay increased from 7 to 22 days, respectively. If assuming a detection delay of 21 days, it was estimated that, for every additional hr of delay, the impact would be an additional approximately 2,000 animals slaughtered and an additional economic loss of $565 million. These findings underline the critical importance that the United States has an effective early detection system in place before an introduction of FMDV if it hopes to avoid dramatic losses to both livestock and the economy.
Development of an Individual Rapid Test Based on Enzymatic Immunofiltration Assay for Detection of Anti–Brucella Abortus Antibody in Bovine Sera
Authors: Genç Oktay , Büyüktanir Özlem , Yurdusev Nevzat
Published: May 2011

To detect bovine antibody directed to smooth lipopolysaccharide (LPS), cell lysate (LYS), O-polysaccharide (OPS), and LPS-deprived chromatographic fractions (ChF) of Brucella abortus, 2 bi-antigenic diagnostic models based on the enzymatic rapid immunofiltration assay (ERIFA), ERIFALPS/LYS and ERIFAOPS/ChF, were developed. Their diagnostic performance was compared with complement fixation test (CFT), Rose Bengal test (RBT), indirect in-house and commercial enzyme-linked immunosorbent assays (iELISA and com-ELISA, respectively), based on the smooth LPS antigen, by using a total of 420 cattle sera collected from aborted-unvaccinated, aborted-unvaccinated and culture-positive, healthy-unvaccinated, and healthy-vaccinated cattle. The results demonstrated excellent agreement and no statistical difference between iELISAs and LPS-, LYS-, OPS-based ERIFA models. However, diagnostic performance of CFT, RBT, and ChF-based ERIFA was less significant than that of LPS-, LYS-, and OPS-based ERIFA models, and iELISAs. The results demonstrated a successful adaptation of the multi-antigenic ERIFA model to anti– B. abortus antibody in bovine sera and suggest that the ERIFA model can be considered as an “individual rapid ELISA” due to its similarity with ELISA, individual applicability, and rapidity in determining reactor animals within 5 minutes. In conclusion, the potential of multi-antigenic applications can make the rapid ERIFA model not only an alternative screening method but also a confirmatory test for bovine brucellosis diagnosis.
Comparison of Detection of Bovine Virus Diarrhea Virus Antigen in Various Types of Tissue and Fluid Samples Collected from Persistently Infected Cattle
Authors: VanderLey Brian , Ridpath Julia , Sweiger Shaun
Published: May 2011

Bovine viral diarrhea viruses are economically important pathogens of cattle. Most infections in susceptible animals are acquired from animals persistently infected with the virus. Surveillance programs rely on skin biopsies of persistently infected (PI) cattle to detect the infection. The purpose of this study was to compare antigen capture enzyme-linked immunosorbent assay (ACE) testing results using different types of samples from PI animals. The intent was to determine comparative detection rates in types of samples that are frequently submitted to diagnostic laboratories for evaluation of cases of unknown etiology or samples that could be easily collected for Bovine viral diarrhea virus (BVDV) screening. Eight types of samples were collected from 40 PI animals. The sample types were ear notches, serum, nasal swabs, conjunctival swabs, oral swabs, rectal swabs, vaginal/preputial swabs, and a tail skin fold biopsy. Each type of sample ( n = 8) for each animal ( n = 40) was evaluated with a commercial ACE kit. When using ACE, tail-skin fold and nasal swab samples were 100% sensitive compared with results using ear notches. Sensitivity using other samples was as follows: serum and vaginal/preputial swabs, 92%; conjunctival swabs, 64%; rectal swabs, 10%; oral swabs, 8%. Testing of tail skin fold biopsies, nasal swabs, and ear notch samples resulted in reliable results. In contrast, other sample types were unreliable for diagnosis of persistent infection in calves.
Isolation of Avian Influenza virus from Polymerase Chain Reaction–Negative Cloacal Samples of Waterfowl
Authors: Zowalaty Mohamed E. El , Abin Martha , Raju Subhatra , Chander Yogesh , Redig Patrick T. , Abd El Latif Hemmat K. , El Sayed Mona A. , Goyal Sagar M.
Published: May 2011

Avian influenza virus (AIV) is one of the most important zoonotic pathogens because of its potential to cause severe disease outbreaks in avian and human hosts. Virus isolation in embryonated chicken eggs (ECEs) remains a gold standard technique for AIV detection. However, some laboratories prefer molecular methods, such as real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), for initial sample screening because of their high throughput sample processing and rapid results. Samples found positive on real-time qRT-PCR are then inoculated in ECEs for virus isolation and characterization. This approach is based on the premise that real-time qRT-PCR will detect all AIV-positive samples. The current study aimed to determine if AIV can be isolated from cloacal samples of waterfowl that were initially found to be negative by real-time qRT-PCR screening. Quantitative RT-PCR–negative cloacal samples (1,369) were inoculated for virus isolation in commercial nonspecific pathogen–free ECEs. After 4 days of incubation, the allantoic fluids were harvested and inoculated in fresh ECEs for a second passage. Allantoic fluids from 147 samples were positive for hemagglutination with chicken erythrocytes. Of the 147 hemagglutination-positive allantoic fluids, 82 were AIV positive when confirmed with real-time qRT-PCR. Ten isolates were subtyped as H7N2 ( n = 7), H7N1, H1N2, and H2N2. In addition, N subtype could be determined for isolates from an additional 25 samples. These results highlight the fact that screening by realtime qRT-PCR may result in some false-negative cloacal samples for AIV.
Isolation of a Variant Porphyromonas Sp. from Polymicrobial Infections in Central Bearded Dragons (Pogona Vitticeps)
Authors: Bemis David A. , Greenacre Cheryl B. , Bryant Mary Jean , Jones Rebekah D. , Kania Stephen A.
Published: May 2011

Isolates of Gram-negative anaerobic bacteria from reptiles have only occasionally been identified to the genus and species level in the veterinary medical literature. In particular, reports identifying Porphyromonas spp. from infections in reptiles are scarce. The present report describes unique Porphyromonas isolates obtained from necrosuppurative infections in central bearded dragons ( Pogona vitticeps). The isolates grew in the presence of oxygen, were strongly hemolytic, and did not produce detectable black, iron porphyrin pigment. Biochemical identification kit numeric biocodes gave high but unreliable probabilities (>99.9%) for identification as Porphyromonas gingivalis. Partial 16S ribosomal RNA gene sequences of the isolates were identical to each other and shared 91% identity with those of Porphyromonas gulae. The isolates may represent a new reptile-associated Porphyromonas species.
Detection of Brucella Canis–Induced Reproductive Diseases in a Kennel
Authors: Gyuranecz Miklós , Szeredi Levente , Rónai Zsuzsanna , Dénes Béla , Dencso László , Dán Ádám , Pálmai Nimród , Hauser Zsófia , Lami Erzsébet , Makrai László , Erdélyi Károly , Jánosi Szilárd
Published: May 2011

Brucella spp. were isolated from an abortion case submitted for laboratory examination 8 months after the first clinical symptoms appeared in a kennel consisting of 31 dogs. Pathological investigations revealed the parallel presence of necrotic placentitis and the strong immunostaining of trophoblast cells by immunohistochemistry (IHC) using hyperimmune rabbit anti- Brucella canis primary antibodies. The rapid slide agglutination test was positive in 7 of 31 (23%) cases. The organism B. canis was successfully cultured from the blood, tissues, or vaginal swabs of only 3 of 31 (10%) cases. The isolated strains were identified as B. canis based on their colony morphology and agglutination with R sera. The strains were initially misidentified as B. suis with the “Bruce-ladder” method, and were subsequently correctly identified as B. canis with a single nucleotide polymorphism (SNP) typing test. Three culture-positive cases and 3 culture-negative cases with histories of reproductive disorders were selected and examined for the presence of B. canis infection using histopathology, IHC, and polymerase chain reaction (PCR) assays. Characteristic histologic lesions were found in all of the 6 animals, whereas IHC and PCR yielded positive results only in single cases from both groups. The results imply that all cases of canine abortion should be examined for brucellosis by bacterial culture of aborted fetuses and placentas. Immunohistochemical examination of placentas is also recommended because it is a quick and sensitive technique compared with bacterial culture. Multiple methods (i.e., serology, blood, and genital bacterial cultures) should be applied simultaneously and repeatedly for the reliable screening of B. canis infection in live individuals.
Piscirickettsia-Like Organisms as a Cause of Acute Necrotic Lesions in Colombian Tilapia Larvae
Authors: Iregui Carlos A. , Vasquez Gersson M. , Rey Alba L. , Verjan Noel
Published: May 2011

Rickettsial organisms are well-known fish pathogens in both natural and culture environments. This study reports an outbreak of disease in red tilapia larvae caused by piscirickettsia-like organisms (PLOs), which lasted from June until October 2009. Severe mortality was recorded almost exclusively in larvae and postlarvae aged 1–22 days old. Although clinical or gross findings were not evident in diseased fish, histopathology revealed severe necrosis of the epidermis and gill epithelium, with concomitant changes in the underlying skeletal muscle as being the most relevant microscopic lesions. Although PLOs were visible with the routine hematoxylin eosin technique, they were better observed with Giemsa and toluidine blue stains. Transmission electron microscopy revealed that the bacterium was located within the cytoplasm and phagolysosoma-like structures of epithelial cells from the gills and the skin. The bacteria measured 0.9 ± 0.2 μm × 2.1 ± 0.6 mm and had a double cell membrane (the outer one having undulating projections), with variable electron-dense and electron-lucent areas. Ultrastructurally, abundant myelin figures surrounded the microorganisms within host cell cytoplasm. Results indicated that Piscirickettsia-like organisms can cause massive epithelial cell damage associated with concomitant alteration of the electrolyte balance.
Lymphoblastic Lymphoma and Leukemic Blood Profile in a Red-Tail Boa (Boa Constrictor Constrictor) with Concurrent Inclusion Body Disease
Authors: Schilliger Lionel , Selleri Paolo , Frye Fredric L.
Published: May 2011

An adult male wild-caught true red-tail boa ( Boa constrictor constrictor), imported from Surinam, was presented for anorexia, extreme lethargy, and coelomic swelling in the cranial third of the body, in the anatomic location of the thymus. The snake died a few minutes after blood sampling via cardiocentesis. Hematology revealed anemia and extreme leukocytosis (820 × 103/ml) characterized by a predominance (95%) of lymphocytes. Necropsy revealed enlargement of most of the visceral organs. Histology confirmed lymphoblastic lymphoma with a leukemic blood profile and diffuse infiltration of some of the heart, thymus, bone marrow, kidney, spleen, lung, and liver. Several large intracytoplasmic eosinophilic inclusion bodies surrounded by narrow clear “halos” were identified within gastric mucosal cells, proximal and distal convoluted tubule epithelial cells, and splenic cells. The final diagnosis was lymphoblast lymphoma with a leukemic blood profile and concurrent inclusion body disease.
Corynebacterium Pseudotuberculosis Secreted Antigen-Induced Specific Gamma-Interferon Production by Peripheral Blood Leukocytes: Potential Diagnostic Marker for Caseous Lymphadenitis in Sheep and Goats
Authors: Rebouças Miriam F. , Portela Ricardo W. , Lima Danielle D. , Loureiro Dan , Bastos Bruno L. , Moura-Costa Lilia F. , Vale Vera L. , Miyoshi Anderson , Azevedo Vasco , Meyer Roberto
Published: May 2011

In the current study, the applicability of the quantification of gamma interferon (IFN-γ) levels for the detection of animals infected with Corynebacterium pseudotuberculosis and for determining caseous lymphadenitis (CLA) clinical status was evaluated. Peripheral blood leukocytes were collected from CLA nonendemic areas animals, from CLA seropositive animals without clinical signs of the disease, and from seropositive animals presenting CLA clinical signs. The leukocytes were stimulated with C. pseudotuberculosis—secreted antigens that were concentrated by the three-phase partitioning technique. An ovine IFN-γ enzyme-linked immunosorbent assay was used to quantify IFN-γ production. Goats and sheep with CLA had higher IFN-γ levels than uninfected seronegative animals. Leukocytes from sheep with CLA chronic abscesses produced higher IFN-γ levels when compared with seropositive sheep without CLA clinical signs, but this difference was not significant in goats. The sensitivity of the assay was 55.8% and 56%, whereas the specificity was 100% and 93%, for goats and sheep, respectively. In conclusion, IFN-γ is a potential marker for the determination of CLA infection status in small ruminants; however, further research is needed to improve assay sensitivity.
Clinical and Morphologic Changes in Ewes and Fetuses Poisoned by Ipomoea Carnea Subspecies Fistulosa
Authors: Armién Aníbal G. , Tokarnia Carlos H. , Peixoto Paulo Vargas , Barbosa Jose D. , Frese Knut
Published: May 2011

Intoxication with Ipomoea carnea has been reported in goats, sheep, and cattle in tropical regions worldwide. The disease has been characterized only in goats; therefore, the present study was conducted in sheep. Nine animals were fed feed rations that contained 3 different concentrations of Ipomoea carnea subsp. fistulosa. Individual intake varied between 10.5 and 135.2 g of fresh plant per kilogram of body weight (BW) per day. Animals first showed clinical signs between day 43 and day 63. The maximum survival time was 133 days. Sheep presented with weight loss and neurologic abnormalities. Neurologic signs were dominated by marked depression, abnormal behavior, and musculoskeletal weakness, with poorly defined motor and proprioceptive deficits. In mature animals, cytoplasmic vacuolation, consistent with accumulation of secondary lysosomes, affected neurons, astrocytes, exocrine pancreatic acinar epithelia, hepatocytes and Kupffer cells, renal tubular epithelia, thyroid follicular epithelia, cortical adrenal epithelia, endothelia and perivascular cells, and macrophages in lymph nodes and spleen. In the central nervous system, there was axonal degeneration and astrogliosis. Abortion was observed as early as day 22 of the trial. In fetal tissues and placenta of chronically poisoned ewes, cytoplasmic vacuolation was histologically detected in neurons, exocrine pancreatic acinar epithelia, hepatocytes, renal tubular epithelia, and thyroid follicular epithelia. All the sheep developed a glycoprotein storage disease, with lysosomal accumulation of N-glycosidically linked oligosaccharides, which was indistinguishable from that induced by the alkaloid swainsonine alone.
Theoretical and Experimental Approaches to Estimate the Usefulness of Pooled Serum Samples for the Diagnosis of Postweaning Multisystemic Wasting Syndrome
Authors: Cortey Martí , Napp Sebastian , Alba Anna , Pileri Emanuela , Grau-Roma Llorenç , Sibila Marina , Segalés Joaquim
Published: May 2011

Classical postweaning multisystemic wasting syndrome (PMWS) diagnosis is based on postmortem findings (histopathology plus viral detection in lymphoid tissues). Because one of the major differences between PMWS-affected and nonaffected pigs is Porcine circovirus-2 (PCV-2) load in serum and tissues, real-time quantitative polymerase chain reaction (qPCR) has been suggested as a potential diagnostic technique for the disease. The objective of the present study was to assess the applicability of qPCR to quantify PCV-2 loads in pooled serum samples as an easy-to-use PMWS diagnostic tool at the herd level. The experimental design included two simulation studies with several serum pool sizes from pigs already screened for PMWS (by histopathology and detection of PCV-2 by qPCR). Several qPCR thresholds were defined and validated with experimental pools created in the laboratory. Quantitative PCR on pooled serum samples did not result in a sufficiently reliable alternate method to the classical PMWS diagnosis method based on individual clinical, histopathological, and PCV-2 detection criteria. However, serum pools seemed to be an alternative at a low economic cost for the quantification of PCV-2 loads in suspicious herds. A targeted (including only clinically diseased animals) sampling approach did not give better estimates compared with a random sampling approach.
Detection of Influenza a Virus in Porcine Oral Fluid Samples
Authors: Detmer Susan E. , Patnayak Devi P. , Jiang Yin , Gramer Marie R. , Goyal Sagar M.
Published: May 2011

Porcine oral fluids have been used for the detection of Porcine reproductive and respiratory syndrome virus and Porcine circovirus-2. The objective of the present study was to determine whether Influenza A virus (FLUAV) is present in porcine oral fluids at detectable levels and to validate a standard FLUAV molecular diagnostic test for porcine oral fluids. Pen-based oral fluid samples were collected on 3, 4, 5, and 6 days postinfection (DPI) from 4 groups of 6 pigs each that were inoculated intratracheally with A/Swine/ Iowa/00239/2004 H1N1 and from 2 untreated or mock-inoculated groups of 6 pigs each that served as negative controls. Individual nasal swabs were also collected from these 36 pigs on 3 and 7 DPI. All oral fluid samples were examined for the presence of FLUAV by matrix gene real-time reverse transcription polymerase chain reaction (real-time RT-PCR) and virus isolation. Nasal swabs were tested initially by virus isolation followed by retest of negative samples with real-time RT-PCR. No oral fluid sample from virus-inoculated pigs was positive by virus isolation, but 15 of 16 positive (94%) oral fluids were positive by real-time RT-PCR. In contrast, virus was isolated from 32 of 48 (67%) nasal swabs collected from virus-inoculated pigs. In addition, 382 of 910 porcine oral fluids collected from pigs in the field between August 1, 2009, and January 31, 2010, were positive by real-time RT-PCR. The results of the present study indicate that pen-based oral fluids provide an easy, effective, and safe collection method for the detection of FLUAV with rapid testing methods such as real-time RT-PCR.
Comparison of RNA Extraction and Real-Time Reverse Transcription Polymerase Chain Reaction Methods for the Detection of Porcine Reproductive and Respiratory Syndrome Virus in Porcine Oral Fluid Specimens
Authors: Chittick Wayne A. , Stensland Wendy R. , Prickett John R. , Strait Erin L. , Harmon Karen , Yoon Kyoung-Jin , Wang Chong , Zimmerman Jeffrey J.
Published: May 2011

The objective of the current study was to evaluate various RNA extraction and polymerase chain reaction (PCR) protocols for the detection of Porcine reproductive and respiratory syndrome virus (PRRSV) in porcine oral fluids. Extraction protocols were selected based on ease of use and compatibility with high-throughput, automated systems. The results showed marked differences among extraction protocols, PCR protocols, and combinations thereof in detecting PRRSV in the oral fluid matrix. An important finding was that PCR reactions were partially inhibited by unknown factors in the oral fluid matrix and that inhibition was reduced by use of a higher concentration of PCR enzymes. The results suggest that further optimization of PCR assays for porcine oral fluids is needed and that laboratories should not assume that methods optimized for detection of virus in serum will perform equally with porcine oral fluids.
Necrotic Enteritis in Young Calves
Authors: Morris Winston E. , Venzano Agustín J. , Elizondo Ana , Vilte Daniel A. , Mercado Elsa C. , Fernandez-Miyakawa Mariano E.
Published: May 2011

Non–enterotoxin (CPE)–producing Clostridium perfringens type A has been associated with enteritis in calves. Recent evidence has suggested that a novel toxin, named beta2 (CPB2), is implicated in the pathogenesis of this disease, although there is little evidence supporting this. In the current study, the role of C. perfringens type A in an outbreak of enteritis in calves was studied. Two 20-day-old dairy calves exhibiting apathy and reluctance to eat, with paresis of the anterior limbs, were euthanized for postmortem examination. Gross and histological changes compatible with acute enteritis, rumenitis, meningitis, and pneumonia were seen in both calves. Clostridium perfringens type A non-CPE, non-CPB2 was isolated from the abomasum and the small intestine. Escherichia coli ONT:H8 (with cdtBIII and f17 virulence genes detected by polymerase chain reaction) was also isolated from the brain, abomasum, and intestine from both calves. All the samples were negative for Salmonella spp. When the C. perfringens strain was inoculated into bovine ligated small and large intestinal loops, cell detachment, erosion, and hemorrhage of the lamina propria were observed, predominantly in the small intestine. The results suggest that non-CPE, non-CPB2 C. perfringens type A is able to induce pathologic changes in the intestine of calves, probably enhanced by other pathogens, such as some pathogenic E. coli strains.
Prevalence and Antimicrogram of Staphylococcus Intermedius Group Isolates from Veterinary Staff, Companion Animals, and the Environment in Veterinary Hospitals in Korea
Authors: Youn Jung-Ho , Yoon Jang Won , Koo Hye Cheong , Lim Suk-Kyung , Park Yong Ho
Published: May 2011

The Staphylococcus intermedius bacterial group (SIG) includes 3 distinct genetically heterogenous species: S. intermedius, S. pseudintermedius, and S. delphini. This pathogen group is associated with many opportunistic skin and ear infections in companion animals. Human infections with S. intermedius and S. pseudintermedius isolates and the emergence of methicillin-resistant isolates have been recently reported, which emphasizes the importance of nationwide identification of SIG isolate prevalence and antibiotic resistance in veterinary clinics. In the present study, a total of 178 SIG isolates were obtained from veterinary staff ( n = 40), companion animals ( n = 115), and the local environment ( n = 23) in 8 Korean veterinary hospitals. Isolates were differentiated into 167 S. pseudintermedius (93.8%) and 11 S. intermedius (6.2%) isolates; S. delphini isolates were not identified. The most effective antibiotics against these isolates included amoxicillin-clavulanic acid, amikacin, nitrofloxacin, imipenem, and vancomycin; whereas ampicillin, penicillin, tetracycline, erythromycin, and trimethoprim-sulfamethoxazole were not effective. Surprisingly, the 128 SIG isolates (71.9%) displayed multiple drug resistance (MDR) against 3 or more antibiotic classes. Out of 52 SIG isolates carrying the methicillin-resistance gene ( mecA), only 34 (65.4%) were oxacillin-resistant, and 49 (94.2%) methicillin-resistant SIG were multidrug resistant. This finding suggests the presence of greater numbers of MDR phenotypes than other isolates ( P < 0.05).
Development of an Immunochromatographic Strip for Serological Diagnosis of Porcine Hemagglutinating Encephalomyelitis Virus
Authors: Chen Keyan , He Wenqi , Lu Huijun , Song Deguang , Gao Wei , Lan Yungang , Zhao Kui , Gao Feng
Published: May 2011

An immunochromatographic strip was developed for the detection of an antibody against Porcine hemagglutinating encephalomyelitis virus (PHEV). Colloidal gold–labeled rabbit anti-pig immunoglobulin G (IgG) was used as the detection reagent, and the PHEV recombinant antigens and goat anti-rabbit IgG were coated on the prototype strip and the control lines, respectively. The immunochromatographic strip was capable of specifically detecting PHEV antibodies in serum with a hemagglutination inhibition (HI) titer of 2 within 10 min. Storage of the strips at room temperature for 6 months or at 4°C for 12 months did not change their sensitivity and specificity. Using HI as a reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 93.41% and 98.42%, respectively. There was a strong agreement between the results obtained by HI and the immunochromatographic strips (κ = 0.926). Additionally, there was a strong agreement between enzyme-linked immunosorbent assay and immunochromatographic strips (κ = 0.929). When the immunochromatographic strip was used for serological diagnosis of 1,117 serum samples in Jilin Province in China, the seropositivity ranged from 6.5% in the Liaoyuan District to 81.6% in the Changchun District. Furthermore, many piglets were seropositive to PHEV, indicating the possible transfer of maternal antibodies via the colostrum. Based on the high specificity, sensitivity, and stability of the immunochromatographic strip, it would be suitable for on-site detection of PHEV antibodies in order to monitor PHEV infections in an animal population.
Occurrence of Chlamydiaceae, Mycoplasma Conjunctivae, and Pestiviruses in Alpine Chamois (Rupicapra r. Rupicapra) of Grisons, Switzerland
Authors: Holzwarth Nathalie , Pospischil Andreas , Mavrot Fabien , Vilei Edy M. , Hilbe Monika , Zlinszky Kati , Regenscheit Nadine , Pewsner Mirjam , Thoma Rudolf , Borel Nicole
Published: May 2011

Because interactions between livestock and chamois occur on Alpine pastures, transmission of infectious diseases is considered possible. Thus, the occurrence of Chlamydiaceae, Mycoplasma conjunctivae, and pestiviruses in Alpine chamois ( Rupicapra r. rupicapra) of the Surselva region (eastern Swiss Alps) was investigated. In total, 71 sera, 158 eye swabs, 135 tissue samples, and 23 fecal samples from 85 chamois were analyzed. The sera were tested by 2 enzyme-linked immunosorbent assay (ELISA) kits specific for Chlamydophila abortus. Eye swabs, tissue, and fecal samples were examined by a Chlamydiaceae-specific realtime polymerase chain reaction (PCR). Positive cases were further investigated by microarray method. One serum sample (1.4%) was positive in 1 of the ELISAs. Eye swabs of 3 chamois (3.8%) were positive for Chlamydiaceae. The microarray method revealed the presence of Chlamydophila abortus, C. pecorum, and C. pneumoniae. All tissue and fecal samples were negative. With real-time PCR, 3.9% of the chamois tested positive for Mycoplasma conjunctivae. One chamois had a simultaneous infection with M. conjunctivae and 2 chlamydial species ( C abortus, C. pecorum). Skin and tongue tissue samples of 35 chamois were negative for pestivirus antigen by immunohistochemistry. It was concluded that in contrast to the findings in Pyrenean chamois ( Capra p. pyrenaica) of Spain, the occurrence of Chlamydiaceae in Alpine chamois of the Surselva region is low, and the transmission between domestic and wild Caprinae seems not to be frequent. Comparably, persistent pestiviral infections do not seem to be common in chamois of the Surselva region.
Validation of Preexisting Reference Intervals: Can the Procedure be Applied to Canine Hemostasis?
Authors: Geffre Anne , Concordet Didier , Trumel Catherine , Braun Jean-Pierre
Published: May 2011

The de novo establishment of reference intervals (RIs) for all variables is beyond the capabilities of many small laboratories. Thus, recent international recommendations propose procedures to adopt RIs established by “donor” laboratories after validation in “receiving” laboratories. The objective of the current study was to use recently published RIs of canine hemostasis tests as possible donor values and evaluate the validation procedure with randomized sets of values obtained in another study of canine RI determination of prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen, and antithrombin (AT). The preanalytical, analytical, and demographic conditions of the donor and receiving laboratories were first compared. To represent new reference individuals, 25 validation sample sets of 20 results of the receiving laboratory were randomly selected for each variable and compared with the RI of the donor laboratory. Validation was rejected in all cases for APTT and AT. Donor RI could be validated in 14 of 25 cases for fibrinogen and in 4 of 25 cases for PT. When preanalytical and analytical differences existed between donor and receiving laboratories, validation procedures consistently rejected preexisting RI. When the differences are smaller, the variability of the results obtained in the validation sample sets tested may be responsible for validations or rejections, which can lead to further misinterpretations of results from patients. Validation of a preexisting reference interval is certainly an interesting option for small laboratories, but progressive determination of the laboratory's own reference interval is probably a better long-term solution.
Prevalence, Sites of Colonization, and Antimicrobial Resistance Among Staphylococcus Pseudintermedius Isolated from Healthy Dogs in Saskatoon, Canada
Authors: Rubin Joseph E. , Chirino-Trejo Manuel
Published: May 2011

In dogs, Staphylococcus pseudintermedius is a common colonizer and is associated with pyoderma, otitis externa, and urinary tract infections. In the current study, nasal, pharyngeal, and rectal swabs were taken from 175 healthy dogs and cultured for S. pseudintermedius. The organism was found in 153 dogs (87.4%), including individuals in which it exclusively colonized in the nares ( n = 1), pharynx ( n = 16), and rectum ( n = 17). Antimicrobial susceptibility testing revealed that a remarkably susceptible population, 46.4% of isolates, was susceptible to all drugs tested, and resistance to penicillin (39.9%) and tetracycline (23.5%) were most common. No methicillin (oxacillin)-resistant isolates were identified. Although 3.3% of isolates were erythromycin resistant, no inducible clindamycin resistance was found. The data provide a baseline for future resistance surveillance and indicate that multiple body sites, including at least the pharynx and rectum, should be tested.
Use of the Intradermal Tuberculin Test in a Herd of Captive elk (Cervus Elaphus Nelsoni) Naturally Infected with Mycobacterium Bovis
Authors: Palmer Mitchell V. , Whipple Diana L. , Payeur Janet B. , Bolin Carole A.
Published: May 2011

In the United States, tuberculosis of captive cervids, caused by Mycobacterium bovis, attracted attention in 1991 when investigations, prompted by the identification of a tuberculous elk ( Cervus elaphus nelsoni) of U.S. origin exported to Canada, revealed tuberculosis in 10 different elk herds in 8 different states. Based on methods used in cattle, official regulations pertaining to testing and eradication of tuberculosis in captive cervids were added to the U.S. Department of Agriculture bovine tuberculosis eradication effort in 1994. However, little published information exists on the accuracy of intradermal tuberculin testing in naturally infected cervids. Evaluation of a captive herd of 71 animals in Wisconsin included postmortem examination and tissue sample collection from both tuberculin test responders and nonresponders. Within this captive herd, of admittedly small size, results showed the single cervical test to have a sensitivity of 88% and a specificity of 69%. Evaluation of diagnostic tests in the species of interest is important, as extrapolation of data obtained from other species may not be appropriate.
Hepatic Yersiniosis Caused by Yersinia Enterocolitica 4:O3 in an Adult Dog
Authors: Byun Jae-Won , Yoon Soon-Seek , Lim Suk-Kyung , Lee O-Soo , Jung Byeong Yeal
Published: May 2011

A 2-year-old Pekinese dog was diagnosed with hepatic yersiniosis. Grossly, white-to-yellow nodules consisting of degenerated inflammatory cells, cell debris, and bacterial clumps were scattered throughout the liver. Histopathologically, suppurative and necrotizing hepatitis was apparent. Yersinia enterocolitica biotype 4, serotype O3 (4:O3) was identified and confirmed in the liver immunohistochemically, using a monoclonal antibody. The virulence genes ystA and ail were detected, but the isolate was negative for autoagglutination and calcium-dependent growth. To confirm systemic yersiniosis in animals, it is imperative that the organism(s) be identified because the hepatic lesions are similar to those of Y. pseudotuberculosis and other diseases, including plague, which is also a zoonotic pathogen.
Sellar Xanthogranuloma in a Dog
Authors: Cramer Sarah D. , Miller Andrew D. , Medici Emily L. , Brunker Jill D. , Ritchey Jerry W.
Published: May 2011

A 7-year-old, intact male standard Poodle dog with hypothyroidism and atypical hyperadrenocorticism developed acute signs of lethargy, weakness, inappetence, vomiting, and diarrhea. Clinical signs progressed to hind limb proprioceptive deficits, aggressive behavior with obtundation, and an equivocal seizure. Necropsy revealed a mass in the sellar region that histologically consisted of multinucleated giant cells, macrophages, and hemosiderin-laden macrophages with fewer lymphocytes and plasma cells admixed with large regions of cholesterol cleft deposition, fibrin, and prominent Rosenthal fibers. Pituitary tissue was not identified on gross or histologic examination, but the mass was partially bordered by epithelial cells. The histologic characteristics are similar to changes described in the human medical literature as xanthogranuloma of the sellar region and xanthogranulomatous hypophysitis.
Paraffin-embedded tissue blot as a sensitive method for discrimination between classical scrapie and experimental bovine spongiform encephalopathy in sheep
Authors: Webb Paul R. , Denyer Margaret , Gough Julie , Spiropoulos John , Simmons Marion M. , Spencer Yvonne I.
Published: May 2011

The paraffin-embedded tissue (PET) blot was modified for use as a tool to differentiate between classical scrapie and experimental bovine spongiform encephalopathy (BSE) in sheep. Medulla (obex) from 21 cases of classical scrapie and 6 cases of experimental ovine BSE were used to develop the method such that it can be used as a tool to differentiate between BSE and scrapie in the same way that differential immunohistochemistry (IHC) has been used previously. The differential PET blot successfully differentiated between all of the scrapie and ovine BSE cases. Differentiation was permitted more easily with PET blot than by differential IHC, with accurate observations possible at the macroscopic level. At the microscopic level, sensitivity was such that discrimination by the differential PET blot could be made with more confidence than with differential IHC in cases where the immunohistochemical differences were subtle. The differential PET blot makes use of harsh epitope demasking conditions, and, because of the differences in the way prion protein is processed in different prion diseases, it can serve as a new, highly sensitive method to discriminate between classical scrapie and experimental BSE in sheep.
Diagnosis of Avian bornavirus infection in psittaciformes by serum antibody detection and reverse transcription polymerase chain reaction assay using feather calami
Authors: de Kloet Arne H. , Kerski Anelle , de Kloet Siwo R.
Published: May 2011

Avian bornavirus (ABV) is the causative agent of proventricular dilatation disease (PDD), a highly devastating and contagious disease of psittacines (parrots and parakeets), which has resulted in the death of many captive birds. Accurate diagnosis of bornavirus infection is therefore important for the identification and isolation of infected birds. The current study showed that nonvascular contour (chest) feather calami provide a ready and minimally invasive source of RNA for the detection of ABV by reverse transcription polymerase chain reaction (RT-PCR). Storage of the feathers at room temperature for at least a month did not affect the results. Serological analysis by enzyme-linked immunosorbent assay (ELISA) showed that identification of anti-bornaviral nucleoprotein P40 antibodies can identify many birds with a past or present infection. The presence of anti-avian bornaviral P24 phosphoprotein and P16 matrix protein antibodies was quite variable, rendering these antibodies less useful for diagnosis of ABV infection. The significance of the present findings is that the use of nonvascular feathers as a source of RNA allows sample collection under conditions where storage of other samples would be difficult. Serum detection by ELISA of anti-P40 antibodies allows the identification of infected birds when RT-PCR fails.
Embryonal carcinoma in the abdominal cavity of a male calf
Authors: Aihara Naoyuki , Yamamoto Naoki , Takagi Takayo , Une Yumi
Published: May 2011

An embryonal carcinoma was diagnosed in the abdominal cavity of a 55-day-old male calf. Macroscopically, a large volume of ascitic fluid was observed along with white to yellowish-white masses of various sizes densely located on the abdominal wall and the surface of abdominal organs. There was an absence of testes in the scrotum. Histologically, the tumor cells were polygonal, and the tumor was mostly composed of sheets of densely packed solid patterns with occasional papillary and tubular structures. Cell nuclei were variable in size, and cellular mitotic rate was high. Immunohistochemically, tumor cells were positive for alpha-fetoprotein, placental alkaline phosphatase, cytokeratin, and carcinoembryonic antigen. Ultrastructurally, the tumor cells had large nuclei, extensive rough endoplasmic reticulum, and small numbers of mitochondria. Microvillus-like structures and desmosomes were occasionally observed. From lectin histochemical examination, the tumor cells were positive for concanavalin A, wheat germ agglutinin, peanut agglutinin, Dolichos biflorus agglutinin, soybean agglutinin, Griffonia simplicifolia I, and Bauhinia purpurea, and negative for Ulex europaeus agglutinin I. Results of histopathological, immunohistochemical, and ultrastructural examinations of the tumor were similar to those obtained for human embryonal carcinoma.
Use of sentinel serology in a Bovine viral diarrhea virus eradication program
Authors: Corbett Erik M. , Grooms Daniel L. , Bolin Steven R. , Bartlett Ben , Grotelueschen Dale M.
Published: May 2011

Forty-seven cattle management groups from 36 herds in a regional Bovine viral diarrhea virus (BVDV) eradication program were selected to evaluate serology as a tool to detect herd infection with BVDV. Serum samples were obtained from 5 non-vaccinated sentinel calves ≥ 6 months old in each management group and virus neutralizing (VN) antibody titers against BVDV genotypes 1 and 2 were determined. A herd was considered positive if 2 or more sentinel calves had VN antibody titers ≥ 128 to either genotype. Results were compared to individual animal testing of all available calves by reverse transcription polymerase chain reaction (RT-PCR) on skin biopsy samples. In 1 management group from 1 herd (n = 24), 3 sentinel calves had VN antibody titers ≥ 128. Three ear notch samples from that herd were positive for BVDV on RT-PCR assay. All other management groups were negative for BVDV. In the present study, the herd sensitivity of sentinel serology was 100% (95% confidence interval [CI]: 0.05–1.0) and herd specificity was 100% (95% CI: 0.90–1.0). The κ value for agreement between sentinel serology and RT-PCR was 1.0 (95% CI: 1.0–1.0). Preliminary results suggest that sentinel animal serology can be utilized in a BVDV eradication program to provide an accurate and efficient evaluation of herd status.
Outbreaks of Eastern equine encephalitis in northeastern Brazil
Authors: Silva Maria L. C. R. , Galiza Glauco J. N. , Dantas Antônio F. M. , Oliveira Rafael N. , Iamamoto Keila , Achkar Samira M. , Riet-Correa Franklin
Published: May 2011

Outbreaks of eastern equine encephalitis observed from May 2008 to August 2009 in the Brazilian states of Pernambuco, Ceará, and Paraíba are reported. The disease occurred in 93 farms affecting 229 equids with a case fatality rate of 72.92%. Main clinical signs were circling, depression or hyperexcitability, ataxia, and progressive paralysis with a clinical manifestation period of 3–15 days. Main histologic lesions were a diffuse lymphocytic encephalomyelitis with neuronal death, satellitosis, neuronophagia, and hemorrhages being more severe in the cerebral gray matter of the telencephalon, diencephalon, and mesencephalon. Some animals also had areas of malacia in the telencephalon, thalamus, and basal nuclei. From 1 case, the virus was isolated by mice inoculation, and in other 13 cases was identified as Eastern equine encephalitis virus by semi-nested reverse transcription polymerase chain reaction. After DNA sequencing, all samples were identified as eastern equine encephalitis through the BLASTn analysis, but samples from the Ceará and Paraíba states corresponded to the same cluster, while the sample from the state of Pernambuco corresponded to a different cluster.