bluuebunny/crossref_metadata_2025 huggingface dataset:
Neutralizing Monoclonal Antibody Sandwich Liquid-Phase Blocking Enzyme-Linked Immunosorbent Assay for Detection of Foot-and-Mouth Disease Virus Type O Antibodies
Authors: Morioka Kazuki , Fukai Katsuhiko , Yoshida Kazuo , Yamazoe Reiko , Onozato Hiroyuki , Ohashi Seiichi , Tsuda Tomoyuki , Sakamoto Kenichi
Published: May 2011

Liquid-phase blocking enzyme-linked immunosorbent assay (LPBE) using the neutralizing monoclonal antibody (mAb) sandwich method (M-LPBE) for detection of Foot-and-mouth disease virus (FMDV) type O antibodies was developed. Two neutralizing mAbs, 72C1 and 65H6, were raised against the FMDV O/JPN/2000 strain, and used as trapping and peroxidase-labeled detecting antibodies, respectively. Sera from animals experimentally infected with FMDV showed specific positive results by M-LPBE, which were correlated with the results of the virus neutralization test (VNT). When 303 negative bovine and 302 negative swine sera were tested, the specificity of M-LPBE was 100% and 99.7%, respectively. In addition, nine samples that had been collected in 2000 in Japan and regarded as evidently false positives by LPBE (supplied by the World Reference Laboratory for Foot-and-Mouth Disease) were uniformly negative by M-LPBE, just like VNT. Therefore, M-LPBE seems to have sufficient specificity for FMDV type O antibody screening and diagnosis.
Identification of Major Antigenic Proteins of Edwardsiella Tarda Recognized by Japanese Flounder Antibody
Authors: Sakai Takamitsu , Matsuyama Tomomasa , Nishioka Toyohiro , Nakayasu Chihaya , Kamaishi Takashi , Yamaguchi Kenichi , Iida Takaji
Published: May 2011

Edwardsiella tarda is a fish pathogen that causes systemic infections in fresh water and marine fish. Determining the antigenic proteins is important for the development of an immunodiagnostic tests and a vaccine for effective infection control in fish. In the current study, antigens were detected by immunoblotting and affinity column chromatography using a Japanese flounder ( Paralichthys olivaceus) antibody produced by experimental infection with E. tarda. GroEL, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), outer membrane protein A, filament protein, 30S ribosomal protein S6, 50S ribosomal protein L9, cold shock protein, and carbon storage protein were identified as antigens of E. tarda through biochemical analyses of the molecular weights, isoelectric points, and N-terminal amino-acid sequences. These proteins can be easily detected in flounder infected with E. tarda and are potential diagnostic markers.
Genotypes, Antibiogram, and Pulsed-Field Gel Electrophoresis Profiles of Escherichia Coli Strains from Piglets in Korea
Authors: Lee Su In , Rayamahji Nabin , Lee Won Jung , Cha Seung Bin , Shin Min Kyung , Roh Yu Mi , Sang Han Yoo
Published: May 2011

Adherence factors and enterotoxins are major virulence factors of enterotoxigenic Escherichia coli (ETEC). Antibiotics have been used frequently for the treatment and prevention of ETEC infection in piggeries worldwide, including Korea. Therefore, data on both virulence profiles and antibiotic resistance patterns are useful in the epidemiological study of ETEC. A total number of 198 E. coli field isolates were examined. The most prevalent pathotype was F1, followed by a combination of F1 and EAST1. All of the 71 isolates were resistant to more than 2 antibiotics used in a disk diffusion test, and 87.94% of the isolates were found to be resistant to more than 4 antibiotics. Investigations were also conducted to correlate the virulence gene profiles with antibiogram and pulsed-field gel electrophoresis (PFGE). Although a high degree of polymorphism was noted among strains having the same virulence patterns, the highest similarity pattern was observed carrying the same virulence profiles and similar antibiogram. Thus, investigation of both virulence profiles and antibiogram is essential to the epidemiological study of ETEC. Moreover, the PFGE method might be applicable as a tool to reveal genetic relatedness among E. coli strains from piggeries in Korea.
Evaluation of a Blocking Enzyme-Linked Immunosorbent Assay for Serological Diagnosis of Bovine Herpesvirus 1
Authors: Godhardt-Cooper Jennifer A. , Zoromski Jeanne , Toohey-Kurth Kathy
Published: May 2011

Bovine herpesvirus 1 (BoHV-1) is an infectious agent of concern in the international export of bovine products; it is endemic in the United States, but it has been eradicated in many countries of the European Union (EU). For export of semen to the EU, accurate assessment of BoHV-1 status of the bull is required and is usually accomplished by measuring the level of antibody to the virus. The gold standard is virus neutralization (VN) using overnight incubation with the virus, a test approved by the World Organization for Animal Health (OIE). Enzyme-linked immunosorbent assay (ELISA) is also approved for international trade. The lone U.S. Department of Agriculture–approved commercial ELISA was compromised with specificity problems, which necessitated the development of a different ELISA. Of 4 monoclonal antibodies evaluated, 1 directed against glycoprotein C of BoHV-1 was found to be the most reliable. One hundred twenty-eight characterized positive samples and 334 negative serum samples were tested. The blocking ELISA showed 97.4% sensitivity and 99.4% specificity as compared with OIE VN. The Wisconsin Veterinary Diagnostic Laboratory ELISA fulfills the OIE requirement for a blocking or competitive ELISA to qualify animals for export to BoHV-1-free countries.
Clinical, Histologic, and Immunohistochemical Features of an Undifferentiated Renal Tubular Carcinoma in a Juvenile Olive Baboon (Papio anubis)
Authors: García Nuria , Ramis Guillermo , Pallarés Francisco J. , Seva Juan I. , Martínez Carlos M. , Quereda Juan J. , Muñoz Antonio
Published: May 2011

An undifferentiated renal tubular carcinoma was diagnosed in a juvenile male olive baboon (Papio anubis). The animal suddenly appeared depressed and refused to eat. During physical examination, a firm, palpable mass in the left abdominal area and flank pain were detected. Clinical pathology findings included mild anemia, hypoalbuminemia, hyponatremia, and mildly increased serum creatinine and urea concentrations. Radiographs revealed a large mass in the left abdominal area. Exploratory laparotomy disclosed a 10 cm × 15 cm multilobulated mass involving the left kidney and adjacent organs. Because of a poor prognosis, the animal was humanely euthanized, and necropsy was performed. Tissue samples of the neoplasm were taken for histopathological examination. Immunohistochemical staining was done using vimentin, cytokeratin, S-100 protein, Ki-67, α-actin, and desmin-specific primary antibodies. Microscopically, elongated and irregular tubules were lined by 2 or more layers of atypical epithelial cells. Anisocytosis, anisokaryosis, and frequent mitotic figures were also observed. Following immunohistochemical staining, the cytoplasm of neoplastic cells was positive for cytokeratin, vimentin, and S-100 protein and negative for α-actin and desmin. Positive nuclear staining for Ki-67 was observed. The neoplasm was diagnosed as an undifferentiated renal tubular carcinoma.
Chronic Copper Toxicosis in Sheep Following the Use of Copper Sulfate as a Fungicide on Fruit Trees
Authors: Oruc Hasan H. , Cengiz Murat , Beskaya Attilla
Published: May 2011

Between January and October 2006, 15 Chios sheep died in a field located near a factory in Orhangazi, Bursa, Turkey. In addition, in May 2007, 2 ewes died after aborting in the same field. Clinical signs in affected animals prior to death were anorexia, hematuria, icterus, incoordination, and ptyalism. Postmortem findings included generalized icterus; yellow, friable livers; distended gallbladders with dense, dark bile; and dark, hypertrophic kidneys with hemorrhage. Copper (Cu) concentrations were measured in multiple specimens of the following: 9 sera, 3 livers, 3 kidneys, 4 plants (including 2 artichoke leaf specimens), 3 soil samples, and 1 drinking water sample. High Cu concentrations were present in the livers, kidneys, and sera of dead sheep, as well as in the vegetation and soil samples from the field. Chronic Cu toxicosis was confirmed as the cause of death attributed primarily to the use of copper sulfate as a fungicide for fruit trees within the field. In addition, factory dust containing Cu might have been an additional factor in the toxicosis.
Intraocular Sarcomas in Two Rabbits
Authors: McPherson Leslie , Newman Shelley J. , McLean Nancy , McCain Stephanie , Vemulapalli Ramesh , Kania Stephen , Dubielzig Richard R.
Published: May 2011

Intraocular neoplasms are described in 2 adult rabbits. The left globe of an 8-year-old male rabbit was enucleated after chronic inflammatory disease resulted in a nonvisual eye. The left globe of a 5-year-old female rabbit also was enucleated after a history of lens-induced uveitis, cataract formation, and resultant glaucoma. In both rabbits, histopathology revealed a variably pleomorphic, poorly differentiated, invasive, intraocular spindle cell neoplasm closely associated with lens and lens capsular fragments. Gram stains failed to detect bacterial organisms or Encephalitozoon cuniculi. Polymerase chain reaction assays, used to amplify the 16S RNA gene of numerous bacteria and E. cuniculi, were also negative. Immunohistochemical staining demonstrated strong, diffuse expression for vimentin; however, staining for smooth muscle actin, cytokeratin, S100, and desmin were negative. Long-standing intraocular inflammation and/or traumatic insults to the eyes were considered as causes of these neoplasms. The histologic features of these intraocular neoplasms closely resemble post-traumatic ocular sarcomas in cats.
Cutaneous Viral Papilloma with Local Extension and Subungual Cyst Formation in a Dog
Authors: Plattner Brandon L. , Hostetter Jesse M.
Published: May 2011

A viral-induced digital cutaneous exophytic papilloma was diagnosed in a 2-year-old, spayed, female Siberian Husky dog with lameness. Digital pain and lameness persisted after removal of the initial papilloma, and the fifth lateral digit was subsequently amputated. Upon histologic examination of the digit, a de novo digital, cutaneous, inverted, viral papilloma and subungual cyst were diagnosed. The inverted cutaneous papilloma, located at the junction of the digital paw pad and ventral nail, extended focally through the nail into the subungual space, where an expansile cyst was formed. Cellular changes suggestive of papillomavirus infection were present in the epithelium of the original exophytic papilloma, as well as the endophytic mass and subungual cyst. Cytopathic effects included ballooning degeneration of keratinocytes, koilocytosis, irregularity of keratohyalin granules, and margination of nuclear chromatin. Numerous faintly basophilic to eosinophilic intranuclear inclusions measuring 10–15 μm in diameter were present within keratinocytes of the exophytic, endophytic, and subungual cystic lesions. Electron microscopy was performed on tissues from all lesions and revealed numerous 40–45 nm diameter hexagonal virions characteristic of papillomavirus that were arranged in crystalline arrays and dense clusters within affected nuclei.
Pleomorphic Leiomyosarcoma in the Hind Leg of a Taiwanese Macaque (Macaca Cyclopis)
Authors: Kim Yongbaek , Kim Hyung-Jin , Cho Doo-Youn , Kang Min-Soo , You Mi-Jin , Kim Dae-Yong
Published: May 2011

An adult, captive Taiwanese monkey ( Macaca cyclopis) presented clinically with a large, rapidly growing mass located in the crus of the left hind leg. The overlying skin was severely ulcerated and necrotic. Radiographs suggested an invasive neoplasm in soft tissue with no bone involvement. The animal's clinical condition progressively worsened over the next 3 months until it died. Necropsy revealed that the mass infiltrated the surrounding skeletal muscle with no evidence of distant metastasis. Microscopically, the mass was highly cellular and composed of round to spindle cells with frequent rhabdoid cells characterized by abundant eosinophilic glassy cytoplasmic inclusions and large, bizarre nuclei. Phosphotungstic acid hematoxylin staining failed to reveal distinct cross-striations within the neoplastic cells. Neoplastic cells were strongly positive for smooth muscle actin and vimentin but were negative for sarcomeric actin, myoglobin, desmin, cytokeratin, S100, and lysozyme. The gross, microscopic, and immunohistochemical findings supported the diagnosis of pleomorphic leiomyosarcoma.
Rhabditid Nematode-Associated Ophthalmitis and Meningoencephalomyelitis in Captive Asian Horned Frogs (Megophrys Montana)
Authors: Imai Denise M. , Nadler Steven A. , Brenner Deena , Donovan Taryn A. , Pessier Allan P.
Published: May 2011

Between 2006 and 2008, 4 captive Asian horned frogs (Megophrys montana) were diagnosed with ocular and neurologic disease associated with rhabditid nematodiasis. Mortality, either spontaneous or by humane euthanasia, was high (3/4, 75%). Gross and histologic findings included varying degrees of ulcerative keratitis, histiocytic uveitis and retinitis, meningoencephalomyelitis, and epidermal chromatophore (iridophore) hyperplasia with intralesional nematodes. Entry into the host was presumed to be by direct invasion of the skin and the cornea with migration through the optic nerve to the brain and spinal cord. One frog was diagnosed with rhabditid nematodiasis antemortem, and clinical signs and lesions in the frog did not progress after unilateral enucleation and anthelminthic treatment were completed. Gross and tissue morphology of the nematodes were consistent with the order Rhabditida. DNA was extracted separately from 2 individual nematodes that were isolated from frozen and ethanol-preserved eye and brain tissue. These DNA templates were used for polymerase chain reaction amplification and sequencing of nuclear 28S large subunit (LSU) and internal transcribed spacer (ITS) ribosomal DNA regions. Comparison of the LSU and ITS sequences to those deposited in GenBank revealed an exact match for Caenorhabditis elegans.
Molecular Detection of Canine Distemper Virus and the Immunohistochemical Characterization of the Neurologic Lesions in Naturally Occurring Old Dog Encephalitis
Authors: Headley Selwyn A. , Amude Alexandre M. , Alfieri Alice F. , Bracarense Ana Paula F. R. L. , Alfieri Amauri A. , Summers Brian A.
Published: May 2011

The current article describes a spontaneous case of old dog encephalitis (ODE) in a 7–year-old, intact, female Miniature Schnauzer dog from Londrina, Paraná, southern Brazil. Unlike conventional distemper encephalomyelitis, ODE is a poorly understood and extremely rare manifestation of Canine distemper virus (CDV) infection. The dog was presented with progressive clinical manifestations consistent with cerebral dysfunction. Briefly, histopathologic lesions were restricted to the forebrain and included chronic multifocal lymphoplasmacytic encephalitis with extensive perivascular cuffing, astrocytosis, and intranuclear inclusions within astrocytes and giant cells, with both intracytoplasmic and intranuclear inclusions. Immunohistochemistry (IHC) was used to identify the antigens of the nucleoprotein (NP) of CDV and to detect cluster of differentiation (CD)3, CD79a, macrophage (MAC) 387, glial fibrillary acidic protein, and vimentin to characterize the neuroparenchymal lesions. By IHC, CDV NP was demonstrated predominantly within neurons and astrocytes. Cells that formed perivascular cuffs and some astrocyte-like cells reacted intensely to vimentin. Reverse transcription polymerase chain reaction assay from brain sections further confirmed a role for CDV in this disease by the amplification and partial sequence analysis of the NP gene. These findings confirmed simultaneous detection of CDV in ODE by IHC and molecular assays. In addition, results of the current study could contribute to the neuropathologic characterization of this rare manifestation of CDV.
Antigen-Capture Blocking Enzyme-Linked Immunosorbent Assay Based on a Baculovirus Recombinant Antigen to Differentiate Transmissible Gastroenteritis Virus from Porcine Respiratory Coronavirus Antibodies
Authors: López Lissett , Venteo Angel , García Marga , Camuñas Ana , Ranz Ana , García Julia , Sarraseca Javier , Anaya Carmen , Rueda Paloma
Published: May 2011

A new commercially available antigen-capture, blocking enzyme-linked immunosorbent assay (antigen-capture b-ELISA), based on baculovirus truncated-S recombinant protein of Transmissible gastroenteritis virus (TGEV) and 3 specific monoclonal antibodies, was developed and evaluated by examining a panel of 453 positive Porcine respiratory coronavirus (PRCoV), 31 positive TGEV, and 126 negative field sera by using another commercially available differential coronavirus b-ELISA as the reference technique to differentiate TGEV- from PRCoV-induced antibodies. The recombinant S protein-based ELISA appeared to be 100% sensitive for TGEV and PRCoV detection and highly specific for TGEV and PRCoV detection (100% and 92.06%, respectively), when qualitative results (positive or negative) were compared with those of the reference technique. In variability experiments, the ELISA gave consistent results when the same serum was evaluated on different wells and different plates. These results indicated that truncated recombinant S protein is a suitable alternative to the complete virus as antigen in ELISA assays. The use of recombinant S protein as antigen offers great advantages because it is an easy-to-produce, easy-to-standardize, noninfectious antigen that does not require further purification or concentration. Those advantages represent an important improvement for antigen preparation, in comparison with other assays in which an inactivated virus from mammalian cell cultures is used.
West Nile Virus Detection in Nonvascular Feathers from Avian Carcasses
Authors: Nemeth Nicole M. , Young Ginger R. , Burkhalter Kristen L. , Brault Aaron C. , Reisen William K. , Komar Nicholas
Published: May 2011

West Nile virus (WNV) is a public health threat and has caused the death of thousands of North American birds. As such, surveillance for WNV has been ongoing, utilizing numerous biological specimens and testing methods. Nonvascular (i.e., fully grown) feathers would provide a simple method of collection from either dead or live birds of all ages and molt cycles, with presumably less biosafety risk compared with other specimen types, including feather pulp. The current study evaluates WNV detection in nonvascular feathers removed from naturally infected avian carcasses of several species groups. Feathers of corvid passeriforms had the highest sensitivity of detection (64%), followed by noncorvid passeriforms (43%), columbiforms (33%), and falconiforms (31%). Storing feathers for 1 year at −20°C or at ambient room temperature resulted in detection rates of infectious WNV of 16% and zero, respectively, but had no effect on detection rates of WNV RNA in a subset of matched feather pairs (47% for both storage temperatures). The efficacy of WNV detection in nonvascular feathers is greatly enhanced by testing multiple feathers. The advantages of using nonvascular feathers over other tissues may outweigh the relatively low detectability of WNV RNA in certain situations such as remote areas lacking resources for acquiring other types of samples or maintaining the cold chain.
Detection and Differentiation of Four Poultry Diseases Using Asymmetric Reverse Transcription Polymerase Chain Reaction in Combination with Oligonucleotide Microarrays
Authors: Tao Qimeng , Wang Xiurong , Bao Hongmei , Wu Jianan , Shi Lin , Li Yanbing , Qiao Chuanling , Yakovlevich Samuilenko Anatolij , Mikhaylovna Poukhova Nina , Chen Hualan
Published: May 2011

Asymmetric reverse transcription polymerase chain reaction (RT-PCR) and microarrays were combined to distinguish 4 viruses, including Avian influenza virus (AIV), Newcastle disease virus (NDV), Infectious bronchitis virus (IBV), and Infectious bursal disease virus (IBDV), and hemagglutinin (HA) subtypes H5, H7, and H9, and neuraminidase (NA) subtypes N1 and N2 of AIV. The AIV matrix protein (M), and HA and NA genes, IBV nucleoprotein (NP) gene, NDV NP gene, and IBDV A fragment gene were cloned into plasmids. These genes were amplified from these positive recombinant plasmids, which included the inserted target genes by PCR. The PCR products were purified and printed on the amino-modified slides as the probes. RNA was extracted from samples and labeled by asymmetric RT-PCR using a cyanine (Cy)3–labeled primers. The labeled complementary (c)DNA was hybridized to the probes immobilized on the glass slides. After hybridization, the microarrays were scanned, and the hybridization pattern agreed perfectly with the known location of each probe. No cross-hybridization could be detected. Results demonstrated that microarray based on asymmetric RT-PCR is an effective way to distinguish AIV, IBV, NDV, and IBDV simultaneously.
Establishment of Reference Intervals for Kaolin-Activated Thromboelastography in Dogs Including an Assessment of the Effects of Sex and Anticoagulant Use
Authors: Bauer Natali , Eralp Oya , Moritz Andreas
Published: May 2011

Tissue factor (TF)- and kaolin-activated thromboelastography (TEG) have been performed in a small number of healthy dogs, but reference intervals have not been assessed in a larger number of dogs. The goal of the current study was to establish reference intervals and assess intra-assay repeatability for kaolin-activated TEG in dogs. Additionally, the impact of sex and the influence of anticoagulant (native blood vs. recalcified citrate anticoagulated blood) were evaluated. Thromboelastography analyses were performed in 56 healthy dogs including German Shepherd Dogs ( n = 19), Beagles ( n = 15), and others ( n = 22). Median age was 2 years (range: 1–6 years) and sex was evenly distributed (31 males and 25 females). To establish reference intervals, citrated whole-blood samples were collected, and TEG was performed 1 hr after sampling. Five TEG variables (R = reaction time; K = clot formation time; α = angle α; MA = maximal amplitude; G-value reflecting clot stability) were evaluated, and reference intervals were defined as the mean + 1.96-fold standard deviation. Intra-assay repeatability was assessed by calculating the pooled variance estimate in duplicate measurements of 6 healthy dogs. The effect of anticoagulant was assessed in 17 specimens. Reference intervals were as follows: R = 1.8–8.6 min; angle α = 36.9–74.6 degrees; K = 1.3–5.7 min; MA = 42.9–67.9 mm, and G = 3.2–9.6 Kdyn/cm 2 . Coefficients of variation for R, K, angle α, MA, and G were 7.6%, 17.7%, 7.4%, 2.9%, and 6.6%, respectively. There was no significant impact of sex or anticoagulant on results. Interindividual variation was higher in native samples than in citrated whole blood. A limitation of the current study was that most of the samples were obtained from Beagles and German Shepherd Dogs. This study provides useful reference intervals for kaolin-activated TEG.
Validation and Application of an Immunofluorimetric Assay for Detection of Serum Free Triiodothyronine and Free Thyroxine Concentrations in Buffalo (Bubalus Bubalis) under Various Physiological Conditions
Authors: Ambrosio Roberto , Sannino Maria Laura , Cortese Laura , Nappi Clemente , Ara Daniela , Cioffi Michele
Published: May 2011

The current study was designed to validate an immunofluorimetric method to measure free triiodothyronine (fT3) and free thyroxine (fT4) concentrations in the serum of buffalo and to develop a better understanding of thyroid metabolism in buffalo ( Bubalus bubalis) of different ages and reproductive statuses. Free triiodothyronine and fT4 were determined in sera from 4 groups of 10 animals, each representing 5–10-year-old lactating buffalo, 2–3–year-old first calf heifers, 1–1.5-year-old virgin heifers, and 5–10–year-old dry buffalo. The average sensitivity of the immunofluorimetric assay was 5.9 pmol/l for fT3 and 14.7 pmol/l for fT4. Intra-assay and interassay coefficients of variation were 5.6% and 5.1% for fT3 and 1.5% and 4.3% for fT4, respectively. Virgin heifers had higher average values for fT3 and fT4, while lactating buffalo had the lowest average values. Dry buffalo and first calf heifers had intermediate fT3 and fT4 values. The findings of this study suggested that lactating buffalo were in a state of low thyroid activity, while virgin heifers had increased thyroid activity.
Assessment of Prothrombin Time, Activated Partial Thromboplastin Time, and Fibrinogen Concentration on Equine Plasma Samples following Different Storage Conditions
Authors: Casella Stefania , Giannetto Claudia , Fazio Francesco , Giudice Elisabetta , Piccione Giuseppe
Published: May 2011

The aim of the present study was to assess the effect of different storage conditions on prothrombin time (PT), activated partial thromboplastin time (aPTT), and fibrinogen concentration in clinical samples from healthy horses. A total of 100 healthy horses of varying breeds and gender, ranging in age from 4 to 18 years, with a mean body weight of 480 + 70 kg, were used. Blood was collected by jugular venipuncture, and a hemochrome-cytometric examination was conducted on all samples. All blood samples were centrifuged and divided into 4 different aliquots to assess clotting parameters by means of a coagulometer. The first aliquots were analyzed 1 hr after collection, the second aliquots were refrigerated at 8°C for 6 hr, the third aliquots were frozen at −20°C for 24 hr, and the fourth aliquots were frozen at −20°C for 48 hr. Significant differences ( P < 0.05) were determined by one-way analysis of variance with repeated measures, and statistical analysis showed a significant effect of the experimental conditions on all parameters studied. In particular, the results demonstrated that coagulation tests can be done within 6 hr when samples are stored at 8°C because the short-term refrigeration does not change the result of analyses; storage at −20°C is acceptable only after 24 hr for PT, aPTT, and fibrinogen measurements because after 48 hr, freezing alters the values of clotting parameters. Therefore, the results of this investigation indicate that clotting parameters remain stable only up to 24 hr in horses without adversely affecting hemostasis test results.
Real-Time Detection of the Mutation Responsible for Progressive Rod-Cone Degeneration in Labrador Retriever Dogs Using Locked Nucleic Acid TaqMan Probes
Authors: Gentilini Fabio , Rovesti Gian Luca , Turba Maria Elena
Published: May 2011

Progressive rod-cone degeneration ( prcd) is a late onset, autosomal recessive, inherited disease in dogs caused by a G > A substitution in the PRCD locus, which has been reported in more than 18 breeds, including Labrador Retriever dogs. In this study, a real-time polymerase chain reaction (PCR) assay, exploiting the features of locked nucleic acid (LNA) fluorescent-labeled probes, was developed to genotype the sequence variants responsible for the disease. Two Labrador Retrievers were diagnosed with prcd by ophthalmological examination performed by a panelist of the Italian hereditary eye disease control program. The 2 dogs, as well as 8 related and 14 unrelated Labrador Retrievers, were genotyped with both direct sequencing of the disease locus and real-time LNA TaqMan PCR assay. Even though the region surrounding the mutation was predicted to be highly structured, making probe annealing difficult, the real-time PCR assay allowed researchers to correctly genotype the dogs in all cases with a sensitivity threshold of 4 ng/reaction of genomic DNA. A real-time PCR assay will allow a high-throughput analysis of a larger cohort of dogs, thereby enabling researchers to investigate the prevalence of the mutated allele in the affected breeds.
Evaluation of a New Commercial Enzyme-Linked Immunosorbent Assay for the Detection of Porcine Antibodies against Trichinella Spp.
Authors: Frey Caroline F. , Buholzer Patrik , Beck Relja , Marinculić Albert , Raeber Alex J. , Gottstein Bruno , Schuppers Manon E.
Published: May 2011

Trichinellosis is a zoonotic disease that is caused by the nematode Trichinella spp. Both European Union regulations and guidelines from the World Organization for Animal Health foresee the possibility of conducting serological surveillance for Trichinella spp. A newly developed commercial enzyme-linked immunosorbent assay (ELISA) was evaluated against 2 existing diagnostic techniques: an in-house ELISA and an in-house Western blot. A total of 875 Trichinella larva-negative samples of pigs and 93 Trichinella larva-positive samples of both naturally and experimentally infected pigs were included in the study. Bayesian modeling techniques were used to correct for the absence of a perfect reference test. The sensitivity and specificity of the commercial ELISA was 97.1–97.8% and 99.5–99.8%, respectively. Sensitivity analysis demonstrated high stability in the models. In a serological surveillance system, ELISA-positive samples should be tested by a confirmatory test. The Western blot is a suitable test for this purpose. With the use of the results of the models, the sensitivity and specificity of a test protocol in both ELISA and Western blot were 95.9% and 99.9%, respectively. The high sensitivity and specificity were achieved with a lower limit of detection than that of the routine artificial digestion test, suggesting that serological surveillance is a valuable alternative in surveillance for Trichinella spp. in pig production.
Antimicrobial Resistance of Streptococci Isolated from Mastitic Bovine Milk Samples in Korea
Authors: Nam Hyang-Mi , Lim Suk-Kyung , Kang Hyun-Mi , Kim Jae-Myoung , Moon Jin-San , Jang Keum-Chan , Joo Yi-Seok , Kang Mun-Il , Jung Suk-Chan
Published: May 2011

The aim of the current study was to determine the antimicrobial resistance of streptococci isolated from mastitic bovine milk samples. A total of 178 isolates belonging to 6 different Streptococcus species were examined: S. uberis (n = 99), S. bovis ( n = 30), S. oralis ( n = 24), S. salivarius ( n = 13), S. intermedius ( n = 7), and S. agalactiae ( n = 5). Only 8.9% (16/178) of the isolates were susceptible to all antimicrobial agents tested in this study, and S. agalactiae and S. intermedius isolates were all resistant to at least 1 antimicrobial agent tested. Overall, the most frequently observed resistance was to tetracycline (61.2%), followed by lincomycin (43.2%), gentamycin (35.3%), oxacillin (34.3%), and erythromycin (28.6%). Cephalothin and penicillin were the only antimicrobial agents to which most of the streptococci (92%) were susceptible. Wide differences in the prevalence of resistance are apparent among the individual species: S. salivarius displayed exceptionally high resistance to cephalothin (23.0%) and oxacillin (76.9%) and S. agalactiae (20%) and S. intermedius (14.2%) to penicillin. Streptococcus salivarius and S. agalactiae were all susceptible to erythromycin, but others showed various rates of resistance ranging from 12.5% to 42.8%. Resistance to 3 or more of 7 antimicrobial agents was observed in all species (37.6%, 67/178).
Disseminated Cutaneous Mast Cell Tumors with Epitheliotropism and Systemic Mastocytosis in a Domestic Cat
Authors: Lamm Catherine G. , Stern Adam W. , Smith Amanda J. , Cooper Emily J. , Ullom Steven W. , Campbell Gregory A.
Published: May 2011

A 15-year-old female Domestic Medium Hair cat presented to the referring veterinarian with a 2-month history of multiple, raised, disseminated, nodular skin lesions. A biopsy of 1 of the lesions was submitted to the Oklahoma Animal Disease Diagnostic Laboratory for evaluation. Histologically, there were multiple dermal nodules composed of sheets of neoplastic round cells. Multifocally, the neoplastic cells formed multiple small clusters of 3–5 cells within the epidermis. Distinct cytoplasmic granules were evident within the neoplastic cells with toluidine blue and Giemsa stains. The neoplastic cells were immunoreactive for c-KIT and lacked immunoreactivity for cluster of differentiation 3 with immunohistochemistry. Based on these findings, multiple epitheliotropic cutaneous mast cell tumors were diagnosed. The cat's health declined rapidly despite aggressive treatment, and the animal was humanely euthanatized. A complete necropsy revealed sheets of similar neoplastic mast cells within the spleen, liver, and individual cells scattered within the bone marrow. Exon 11 of the c-KIT messenger RNA from 1 of the cutaneous masses and the spleen was amplified with reverse transcription polymerase chain reaction, sequenced, and compared with the published c-KIT messenger RNA sequence from fetal cat tissues. The maximum identity was 100% for both tissue samples. To the authors’ knowledge, the present report is the first to describe disseminated cutaneous mast cell tumors with epitheliotropism and systemic mastocytosis in a domestic cat.
Postmortem Diagnostic Investigation of Disease in Free-Ranging Marine Turtle Populations: A Review of Common Pathologic Findings and Protocols
Authors: Flint Mark , Patterson-Kane Janet C. , Limpus Colin J. , Work Thierry M. , Blair David , Mills Paul C.
Published: May 2011

Over the past few decades, there have been increasing numbers of reports of diseases in marine turtles. Furthermore, in recent years, there have been documented instances of apparently new diseases emerging in these species of which the etiology and/or pathogenesis remain unknown. These instances i) raise concern for the survival of marine turtles, and ii) question the health and stability of the benthic marine environments in which turtles live. Knowledge of common disease processes and pathologic changes in lesions, along with a standardized approach to postmortem and sample collection are required to document and understand the host-agent-environment interactions in marine turtle health. This review combines, for the first time, a standardized approach to the postmortem of marine turtles for veterinary clinicians, with a concurrent descriptive review of the gross and microscopic pathologic changes in lesions commonly seen.
Phenotypic and Genetic Characterization of Pasteurella Multocida and Related Isolates from Rabbits in Switzerland
Authors: Stahel Anina B. J. , Hoop Richard K. , Kuhnert Peter , Korczak Bożena M.
Published: May 2011

Several bacteria belonging to the family Pasteurellaceae are potential pathogens in rabbits. In particular, Pasteurella multocida is considered to be important, and outbreaks caused by this species result in considerable economic losses in rabbitries. However, Pasteurellaceae spp. isolated from rabbits are poorly characterized, and thus, proper identification of P. multocida isolates from these animals is problematic and often unsatisfactory, thereby hampering epidemiological investigations. Therefore, 228 isolates from rabbit populations originating from a breeding and fattening organization with group management and postmortem cases with pasteurellosis from individual owners were phenotypically and genotypically analyzed using biochemical tests and repetitive extragenic palindromic polymerase chain reaction (REP-PCR). Furthermore, 41 samples representing observed phenotypes were selected for phylogenetic analysis using 16S ribosomal RNA and rpoB genes. The REP-PCR typing and phylogenetic analyses correlated well and appeared to be distinct molecular methods for characterization of rabbit isolates. Phenotyping, however, diverged from molecular recognition, reflecting the problematic conventional diagnosis of these strains. The fermentation of sorbitol appeared to be an imprecise indicator for P. multocida subspecies classification. According to REP-PCR and sequencing results, 82% of the isolates were characterized as P. multocida subsp. multocida, 3% as P. multocida subsp. septica, and 5% as P. multocida. Further, 5% were identified as Pasteurella canis. The other 5% represented a homogeneous group of unknown species belonging to the Pasteurellaceae. Samples obtained from individual postmortem cases demonstrated a higher phenotypic and genetic heterogeneity than samples from group management rabbits.
Cytokeratin and Vimentin Co-Expression in 21 Canine Primary Pulmonary Epithelial Neoplasms
Authors: Burgess Hilary J. , Kerr Moira E.
Published: May 2011

Co-expression of cytokeratin and vimentin has been traditionally associated with a few select tumors. However, this phenomenon is being recognized in a wider range of tumors. Twenty-one canine primary pulmonary epithelial neoplasms were evaluated for the co-expression of cytokeratin and vimentin. The histologic pattern and grade, and an immunohistochemical grade for cytokeratin and vimentin staining, were determined for each neoplasm. Adenocarcinomas predominated, and histologically, most tumors were grade II. All of the neoplasms stained positive for cytokeratin, while only 8 (38%) stained positive for both vimentin and cytokeratin. Papillary adenocarcinomas were consistently vimentin negative. The anaplastic histologic pattern had significantly more vimentin staining than the other histologic patterns. There was no significant difference in histologic grade or grading criteria between those tumors that stained with vimentin and those that did not. The present study established that cytokeratin and vimentin co-expression occurs in canine primary pulmonary epithelial tumors at a similar frequency to human pulmonary neoplasms. Further investigation will be needed to characterize the significance of this finding, particularly with respect to prognosis.
A Retrospective Immunohistochemical Study Reveals Atypical Scrapie has Existed in the United Kingdom since at Least 1987
Authors: Webb Paul R. , Powell Linda , Denyer Margaret , Marsh Sarah , Weaver Colin , Simmons Marion M. , Johns Elizabeth , Sheehan John , Horsfield Peter , Lyth Chris , Wilson Christina , Long Ann , Cawthraw Saira , Saunders Ginny C. , Spencer Yvonne I.
Published: May 2011

Atypical scrapie is a relatively recent discovery, and it was unknown whether it was a new phenomenon or whether it had existed undetected in the United Kingdom national flock. Before 1998, the routine statutory diagnosis of transmissible spongiform encephalopathy (TSE) in sheep relied on the presence of TSE vacuolation in the brainstem. This method would not have been effective for the detection of atypical scrapie. Currently, immunohistochemistry (IHC) and Western blot are commonly used for the differential diagnosis of classical and atypical scrapie. The IHC pattern of PrP d deposition in atypical scrapie is very different from that in classical scrapie using the same antibody. It is thus possible that because of a lack of suitable diagnostic techniques and awareness of this form of the disease, historic cases of atypical scrapie remain undiagnosed. Immunohistochemistry was performed on selected formalin-fixed, paraffin-embedded (FFPE) blocks of ovine brain from the Veterinary Laboratories Agency archives that were submitted for various reasons, including suspect neurological disorders, between 1980 and 1989. It was found that PrP d deposits in a single case were consistent with atypical scrapie. A method was developed to obtain a PrP genotype from FFPE tissues and was applied to material from this single case, which was shown to be AHQ/AHQ. This animal was a scrapie suspect from 1987, but diagnosis was not confirmed by the available techniques at that time.
Rapid Detection of Infectious Bursal Disease Virus by Reverse Transcription Loop-Mediated Isothermal Amplification Assay
Authors: Xue Chunyi , Zhang Yun , Zhou Qingfeng , Xu Cong , Li Xiaoming , Cao Yongchang
Published: May 2011

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the rapid identification of Infectious bursal disease virus (IBDV). The RT-LAMP assay used a set of 4 primers to amplify the viral protein 2 gene of IBDV for the detection of IBDV, showing not only high efficiency but also analytic specificity. The data demonstrated that the RT-LAMP assay detected 30 different IBDV isolates, had no cross-reaction with 3 other avian viruses ( Infectious bronchitis virus, Newcastle disease virus, and Avian influenza virus), and obtained a 95.45% sensitivity in 22 positive clinical samples in reference to virus isolation. Therefore, this rapid, specific, sensitive, and convenient RT-LAMP assay could be applicable to the identification of IBDV in less-equipped laboratories as well as in the field.
Molecular Typing of Staphylococcus Aureus Strains from Ovine Mastitis by Pulsed-Field Gel Electrophoresis and Polymerase Chain Reaction Based on Coagulase and Protein A Gene Polymorphisms
Authors: Çiftci Alper , Onuk E. Emek , Findik Arzu , Yildirim Tuba , Sogut Mehtap Unlu
Published: May 2011

Staphylococcus aureus is one of the most important etiologic agents of ovine mastitis. To develop effective control measures for mastitis, it is important to type S. aureus strains that have considerable genetic heterogeneity. In the current study, 47 S. aureus strains isolated from ovine mastitis were typed by polymerase chain reaction (PCR) based on coagulase ( coa) and protein A ( spa) polymorphisms and by pulsed-field gel electrophoresis (PFGE). Eight different coa types and 4 spa types were identified by PCR. While the most prevalent coa type was CG2 (42.56%), the spa types S4 and S1 were the most commonly observed (44.68% and 38.29%, respectively). Nineteen different pulsotypes were identified, and 12 of these were represented by a single isolate. Pulsotypes J and K were predominant and each represented 9 isolates (19.14%). All isolates belonging to J and K pulsotypes were CG2. Although all 9 isolates belonging to the J pulsotype were S4, all isolates in the K pulsotype were S1. While PFGE was found to be the best discriminatory technique for distinguishing strains, coa and spa types were found to be in correlation with PFGE types and can be used for quick, preliminary epidemiologic studies for detecting strains that may cause mastitis.
Isolation Rates, Serovars, and Toxin Genotypes of Nicotinamide Adenine Dinucleotide-Independent Actinobacillus Pleuropneumoniae among Pigs Suffering from Pleuropneumonia in Spain
Authors: Maldonado Jaime , Valls Laura , Martínez Eva , Riera Pere
Published: May 2011

Actinobacillus pleuropneumoniae is the etiologic agent of swine pleuropneumonia, a major production-limiting disease in the pig industry. In the current study, 2,171 lung specimens obtained from pigs housed in 870 Spanish pig farms in regions of substantial pig production were examined. Conventional microbiology, coupled with species-specific polymerase chain reaction, identified 127 biovar 2 isolates, accounting for 25.3% of all A. pleuropneumoniae ( n = 502) detected. Most isolates (79%) were recovered as pure primary cultures or as the predominant bacteria from lungs exhibiting lesions typical of acute swine pleuropneumonia. Coagglutination testing identified the isolates as belonging to serovars 2 (4.7%), 4 (4.7%), 7 (68.5%), and 11 (1.6%); however, 26 isolates were nontypeable. All biovar 2 isolates showed genes of the apxII operon alone, which encodes the corresponding ApxII exotoxin, leading to a different gene pattern for isolates in serovars 2, 4, and 11 compared with those of biovar 1. From this survey, it can be concluded that A. pleuropneumoniae biovar 2 infections are common in pigs in Spain, and they may be a common cause of respiratory disease in swine.
Species Distribution and Resistance Patterns to Growth-Promoting Antimicrobials of Enterococci Isolated from Pigs and Chickens in Korea
Authors: Hwang In Yeong , Ku Hyun Ok , Lim Suk Kyung , Park Choi Kyu , Jung Gab Su , Jung Suk Chan , Nam Hyang Mi
Published: May 2011

A total of 147 Enterococcus faecium and 165 Enterococcus faecalis isolates from fecal samples of chickens and pigs at slaughterhouses in Korea were tested for their resistance to 8 growth-promoting antimicrobials commonly used in animals and quinupristin and dalfopristin. Resistance to most antimicrobials was very common among both E. faecalis and E. faecium. In particular, E. faecalis showed almost no susceptibility to all the antimicrobials tested except penicillin and flavomycin, to which 1.4% and less than 24% showed resistance, respectively. Although the prevalence of resistance was lower than in E. faecalis, E. faecium showed relatively uniform resistance to all the agents tested. Among the antimicrobials tested, virginiamycin and penicillin were the most effective against E. faecium isolates: less than 31% and 41% showed resistance to those 2 antimicrobials, respectively. Penicillin was the only agent that showed relatively strong activity against both E. faecalis and E. faecium. Resistance observed in E. faecalis and E. faecium against most antimicrobials used for growth promotion was more prevalent in Korea than in European countries. The current study is the first report of resistance against feed additive antimicrobials in enterococcal isolates from livestock in Korea.
Comparison of Conventional Direct and Enrichment Culture Methods for Erysipelothrix spp. from Experimentally and Naturally Infected Swine
Authors: Bender Joseph S. , Kinyon Joann M. , Kariyawasam Subhashinie , Halbur Patrick G. , Opriessnig Tanja
Published: May 2011

The objective of the current study was to compare the diagnostic performance of a direct isolation method for Erysipelothrix spp. with a broth-based enrichment technique. Samples were obtained from three sources: 1) experimentally inoculated pigs, 2) porcine tissue samples submitted to the Iowa State University Veterinary Diagnostic Laboratory (Ames, IA), and 3) tissues from condemned carcasses at an abattoir. Culture plates from direct isolation and broth-based technique were evaluated for growth at 24 and 48 hr. Results indicated that the broth enrichment method was markedly more sensitive for the isolation of Erysipelothrix spp. To the authors' knowledge, this is the first comparison of direct culture and broth-based enrichment methods for the isolation of Erysipelothrix spp. Interestingly, in several samples, a Gram-positive bacterium with almost identical growth characteristics to Erysipelothrix spp. was detected and identified as a Vagococcus sp. through 16S ribosomal RNA gene sequencing. The results of this study indicate that the broth-based enrichment method should be used for the isolation of Erysipelothrix spp. from clinical samples with a history suggestive of erysipelas and that Vagococcus spp. is potentially an important differential diagnosis.
Adenomyofibroma of the Fimbria in a Cynomolgus Monkey (Macaca Fascicularis)
Authors: Starost Matthew F.
Published: May 2011

A 20-year-old female cynomolgus monkey ( Macaca fascicularis) was presented for an end-of-study necropsy. At necropsy, a 2 cm × 1.5 cm × 1 cm, butterfly-shaped, multilobulated mass extended off the right uterine tube fimbria. Microscopically, the mass was composed of large, plump, finger-like projections lined primarily by simple columnar ciliated epithelium. The interstitium contained a proliferation of smooth muscle stromal cells admixed with varying amounts of collagen. A diagnosis of adenomyofibroma of the fimbria was made. This benign neoplasm should be considered as a differential diagnosis for masses arising from the fallopian tube in old-world macaques.