bluuebunny/crossref_metadata_2025 huggingface dataset:
A cultural-existential approach to therapy: Merleau-Ponty’s phenomenology of embodiment and its implications for practice
Authors: Felder Andrew J. , Robbins Brent Dean
Published: May 2011

Maurice Merleau-Ponty’s phenomenological philosophy provides the basis for a form of cultural-existential therapy. Through an examination of Merleau-Ponty’s analysis of the phantom limb and anosognosia, we develop a cultural-existential approach to “psychopathology” and its treatment. In the course of this analysis, ego-syntonic labels are seen in the light of culture-syntonic considerations, depth analysis is married to breadth analysis, empathic understanding is re-understood through a dialectical mode of understanding, medical and psychological analyses are recast within a cultural analysis, and being is resituated within a flesh ontology. Whereas a cultural-existential psychotherapy may compassionately rally around a therapy of situated individuals, it also calls for mindful attention to a therapeutics of culture.
Follow-up of lupus choroidopathy with optical coherence tomography
Authors: Ozturk BT , Bozkurt B , Karademir Z , Kerimoglu H
Published: May 2011

Abstract: A 36-year-old female followed with the diagnosis of systemic lupus erythematosus complained of bilateral visual loss. Ophthalmological examination revealed disc edema with irregular borders, edematous and pale retina with widespread cotton wool spots, intraretinal hemorrhages and serous retinal detachment in both eyes. Optical coherence tomography (OCT) scans showed intraretinal and subretinal fluid creating cystic cavities with central subfield macular thickness values of 994 and 912 µm in the right and left eye, respectively. The follow-up scans after the treatment with systemic steroid, acetylsalicylic acid and cyclosporine documented resolution of this fluid accumulation and a decrease in macular thickness measurements. These clearly demonstrated that OCT, which is a fast, objective and non-invasive technology, may be an adjunctive imaging tool for the diagnosis and follow-up of lupus choroidopathy.
Coinfection by Porcine Circoviruses and Porcine Parvovirus in Pigs with Naturally Acquired Postweaning Multisystemic Wasting Syndrome
Authors: Ellis J. A. , Bratanich A. , Clark E. G. , Allan G. , Meehan B. , Haines D. M. , Harding J. , West K. H. , Krakowka S. , Konoby C. , Hassard L. , Martin K. , McNeilly F.
Published: May 2011

Postweaning multisystemic wasting syndrome (PMWS) is an emerging disease in swine. Recently, the disease has been reproduced with inocula containing a newly described porcine circovirus (PCV), designated PCV 2, and porcine parvovirus (PPV). In order to determine if these viruses interact in naturally acquired PMWS, affected tissues from field cases were examined by immunohistochemistry (IHC) and polymerase chain reaction (PCR) for PCV 2 and PPV, as well as by PCR for the other recognized porcine circovirus, PCV 1. Porcine circovirus 2 was detected by PCR or IHC in affected fixed or frozen tissues from 69 of 69 cases of PMWS collected over 3 years from 25 farms. Porcine parvovirus was detected in 12 of the same cases, and PCV 1 was detected in 9 of 69; however, an apparent decrease was found in the sensitivity of the PCRs used to detect the latter 2 viruses when fixed tissue from the same cases were compared with the use of frozen tissues. Porcine circovirus 2 was not detected by PCR in affected tissues from 16 age-matched pigs that had Streptococcus suis-associated disease. Electron microscopic examination of plasma pooled from 15 pigs with PMWS revealed the presence of PCV and PPV, whereas these viruses were not observed in pooled plasma from 5 age-matched clinically normal pigs. These results confirm and extend previous findings documenting a consistent association of PCV 2 with PMWS. As well, infection by PPV or PCV 1 or both may be an important cofactor in the pathogenesis of some, but apparently not all, cases of PMWS.
Porcine Postweaning Multisystemic Wasting Syndrome in Korean Pig: Detection of Porcine Circovirus 2 Infection by Immunohistochemistry and Polymerase Chain Reaction
Authors: Choi Changsun , Chae Chanhee , Clark Edward G.
Published: May 2011

This report describes the first diagnosis of porcine circovirus (PCV) infection in weaned pigs with postweaning multisystemic wasting syndrome in Korea by immunohistochemistry and polymerase chain reaction. The most unique lesions were multifocal granulomatous inflammation affecting lymph nodes, liver, and spleen, characterized by infiltrates of epithelioid macrophages and multinucleated giant cells. Circoviral antigen was detected in formalin-fixed sections and was usually present in large, round, dendritic cells in the white pulp of spleen and remnants of follicles in lymph nodes. Lymphoid follicles in the tonsils also contained PCV antigen. A 530–bp DNA fragment of circovirus was successfully amplified from all tested lymph nodes, liver, and spleen.
Nonsuppurative Meningoencephalitis Associated with Aleutian Mink Disease Parvovirus Infection in Ranch Mink
Authors: Dyer N. W. , Ching B. , Bloom M. E.
Published: May 2011

Severe nonsuppurative meningoencephalitis associated with Aleutian mink disease parvovirus (ADV) infection was observed in adult ranch mink. Brain lesions included severe, locally extensive to coalescing lymphoplasmacytic meningoencephalitis with accompanying gliosis, satellitosis, and mild extension of inflammation into the leptomeninges. ADV was identified in mesenteric lymph node, spleen, brain, and liver of affected mink by polymerase chain reaction techniques. Sequences of the ADV isolate (TH5) revealed 2 unique residues in the region of the viral genome that determines pathogenicity. These findings suggest that certain strains of ADV may preferentially cause disease in the nervous system. ADV infection should be considered in the differential diagnosis of neurologic disorders in mink.
Type C Botulism in Dairy Cattle from Feed Contaminated with a Dead Cat
Authors: Galey F. D. , Terra R. , Walker R. , Adaska J. , Etchebarne M. A. , Puschner B. , Fisher E. , Whitlock R. H. , Rocke T. , Willoughby D. , Tor E.
Published: May 2011

Four hundred twenty-seven of 441 adult Holstein dairy cattle from a 1,200-cow dairy died over a 1-week period during early spring 1998. Affected animals were from 4 late lactation pens, one of which included the bull string. Signs included weakness, recumbency, watery diarrhea, and death. Eighty animals from the 4 pens were dead approximately 8 hours after the first ill cows were noted. Affected cows would collapse on stimulation and extend all 4 limbs with moderate rigidity. Several lacked lingual tonus and had abdominal breathing patterns. The animals had been fed a load of total mixed ration that included a rotten bale of oat hay containing a dead cat. No common toxicants were identified, and pathologic examination revealed no consistent lesions. Testing of tissue from the cat carcass found in the feed sample using mouse protection bioassay identified the presence of type C botulinum toxin. Samples of feed, tissue from affected animals, cat tissue from feed, milk, and serum were also tested using an enzyme-linked immunosorbent assay (ELISA) specific for type C botulinum. Two samples of rumen contents were tested and found to be positive for botulism by ELISA, and 1 of 3 liver samples had a weak positive finding. No botulinum toxin was found in milk or sera using the ELISA.
Urinary Alkaloid Excretion as a Diagnostic Tool for Fescue Toxicosis in Cattle
Authors: Hill N. S. , Thompson F. N. , Stuedemann J. A. , Dawe D. L. , Hiatt E. E.
Published: May 2011

Fescue toxicosis research studies have often included serum prolactin as a physiologic index of the disorder. Serum prolactin has not been used as a clinical measure of fescue toxicosis because of variation associated with sex and physiologic condition of the animal and climatic and seasonal factors. The primary excretory route of the alkaloids responsible for this toxicosis is the urine. Three pasture experiments were conducted to examine serum prolactin and urinary ergot alkaloid variability among steers continuously grazing endophyte-infected (E+) or endophyte-free (E-) tall fescue and among steers that were switched from one pasture form to the other. A fourth grazing experiment was used to examine how to best to manage the steers prior to sampling for urinary ergot alkaloid excretion. Coefficients of variability for urinary alkaloid excretion were consistently lower (46–65%) than serum prolactin (64–142%). Urinary alkaloid excretion patterns changed within 12 hours following switching steers from E+ to E- pasture or visa versa, but serum prolactin was recalcitrant to change. Because it is less variable and more dynamic than serum prolactin, urinary alkaloid excretion can be used for health assessment of steers grazing E+ and E- pastures. Regression analysis established a quadratic relationship between alkaloid excretion and average daily weight gain, with a regression coefficient of 0.86. Urinary alkaloid analysis was useful in determining whether cattle were consuming toxic tall fescue.
Detection of Streptococcus Suis by in Situ Hybridization, Indirect Immunofluorescence, and Peroxidase-Antiperoxidase Assays in Formalin-Fixed, Paraffin-Embedded Tissue Sections from Pigs
Authors: Boye Mette , Feenstra Anne A. , Tegtmeier Conny , Andresen Lars Ole , Rasmussen Søren R. , Bille-Hansen Vivi
Published: May 2011

Streptococcus suis is an important pathogen in pigs and is considered a zoonotic agent. To aid diagnosis of infection caused by S. suis, a species-specific probe targeting 16S ribosomal RNA was designed and used for fluorescent in situ hybridization. Two additional immunohistochemical detection methods, an indirect immunofluorescence assay and a peroxidase-antiperoxidase method, using polyclonal antibodies also were developed. The specificity of the oligonucleotide probe was examined by whole-cell and dot-blot hybridization against reference strains of the 35 serotypes of S. suis and other closely related streptococci and other bacteria commonly isolated from pigs. The probe was specific for S. suis serotypes 1–31. The specificity of the polyclonal antibodies, which has previously been evaluated for use in diagnostic bacteriology for typing of serotype 2, was further evaluated in experimentally infected murine tissue with pure culture of different serotypes of S. suis, related streptococci, and other bacteria commonly found in pigs. The polyclonal antibodies against S. suis serotype 2 cross-reacted with serotypes 1 and 1/2 in these assays. The in situ hybridization and the immunohistochemical methods were used for detection of S. suis in formalin-fixed, paraffin-embedded tissue sections of brain, endocardium, and lung from pigs infected with S. suis. The methods developed were able to detect single cells of S. suis in situ in the respective samples, whereas no signal was observed from control tissue sections that contained organisms other than S. suis. These techniques are suitable for determining the in vivo localization of S. suis for research and diagnostic purposes.
Assessment of the Long-Term Shedding Pattern of Salmonella Serovar Choleraesuis Following Experimental Infection of Neonatal Piglets
Authors: Anderson Robin C. , Genovese Ken J. , Harvey Roger B. , Stanker Larry H. , DeLoach John R. , Nisbet David J.
Published: May 2011

In the United States, swine salmonellosis is most often attributed to infections by Salmonella serovar choleraesuis. As a host-adapted pathogen rarely found in nonswine sources, S. choleraesuis is thought to be spread primarily via horizontal transmission, with carrier animals playing an important role. Little has been reported regarding infection of neonatal piglets, particularly regarding their potential to become carriers. Evidence reported herein demonstrates that piglets experimentally infected by S. choleraesuis at 2 days of age were capable of shedding the pathogen for up to 85 days postinfection, at which time the study was concluded. This study also presents findings supporting the use of GN-Hajna as a preenrichment medium for the isolation of S. choleraesuis.
Representational Differential Analysis Detects Amplification of Satellite Sequences in Postweaning Multisystemic Wasting Syndrome of Pigs
Authors: Bratanich Ana C. , Ellis John A. , Blanchetot Alain
Published: May 2011

Representational difference analysis (RDA) was used as a molecular approach to identify unique sequences associated with postweaning multisystemic wasting syndrome (PMWS) in pigs. Three rounds of subtractive hybridization and amplification between driver DNA extracted from normal pigs and tester DNA from PMWS-affected animals were performed. The final product corresponding to sequences associated with PMWS in pigs was analyzed using agarose gel electrophoresis, and 9 fragments were visualized after staining with ethidium bromide. Eight recombinants were successively cloned and sequenced, and the results were then compared with existing databases. Most of the PMWS clones isolated were satellite sequences from pig centrometric regions and 1 was a microsatellite sequence. One clone represented a microsatellite sequence, and 2 clones showed no homology with any gene found in the databases. The sequence comparison data did not reveal any homology with an infectious agent such as a virus or a bacterium. In the present experimental setting, it was concluded that PMWS in pigs triggers molecular changes such as an amplification of genomic regions containing repeated sequences.
Analysis by Sodium Dodecyl Sulfate Polyacrylamide gel Electrophoresis and Western Blot of Nonspecific and Specific Viral Proteins Frequently Detected in Different Antigen Preparations of Bovine Leukemia Virus
Authors: Llames Louie , Gómez-Lucia Esperanza , Doménech Ana , Suárez Guillermo , Goyache Joaquín
Published: May 2011

Bovine leukemia virus (BLV) infection in cattle is seldom manifested clinically, and is routinely diagnosed by serologic tests such as enzyme-linked immunosorbent assay or Western blot (WB). Because of the difficulty in interpreting WB results, the aim of the present study was to determine which of the bands observed in WB were specifically produced by BLV and which corresponded to nonspecific proteins, either derived from medium components or of a cellular nature. Five different BLV antigen preparations from 2 cell lines (FLK-BLV and BLV-bat2) frequently used for the production of BLV antigen were compared. The protein profiles of these antigen preparations were analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis and WB. Fetal calf serum, required for cellular growth and important in induction of viral transcription in vitro, was identified as a source of irrelevant proteins. In this study, 15 nonspecific protein bands in the growth medium were observed. These bands interfered with the interpretation of results. A nonspecific protein (25 kD) that was highly reactive in cell lysate preparation from BLV-bat2 was also detected. The unequivocal identification of protein bands, both specific and nonspecific, seen in WB is important not for understanding the protein profile of antigen preparations but also for determining if an animal is BLV positive or negative.
Effects of Storage Conditions and Hemolysis on Vitamin E Concentrations in Porcine Serum and Liver
Authors: Hooser Stephen B. , McCarthy Jennifer M. , Wilson Christina R. , Harms Jennifer L. , Stevenson Gregory , Everson Robert J.
Published: May 2011

Vitamin E (α-tocopherol) is an antioxidant vitamin important in protecting unsaturated fatty acids in lipid membranes from peroxidation. Variation in collection, storage, and shipping conditions of samples can potentially lead to breakdown of vitamin E prior to analysis. Therefore, the purposes of this project were 1) to determine the stability of vitamin E in refrigerated and frozen porcine liver and serum and 2) to evaluate the effects of red blood cell (RBC) hemolysis on porcine serum vitamin E concentrations. Porcine liver and nonhemolyzed serum were collected and stored refrigerated or frozen. Samples were analyzed for vitamin E immediately or on days 2, 3, 7, or 14. In addition, porcine RBCs were added to normal serum at concentrations from 1 × 106 to 1 × 109 RBC/ml and hemolyzed by freeze-thaw prior to analysis for vitamin E or products of lipid peroxidation.
Fatal Pulmonary Edema in White-Tailed Deer (Odocoileus Virginianus) Associated with Adenovirus Infection
Authors: Sorden Steven D. , Woods Leslie W. , Lehmkuhl Howard D.
Published: May 2011

Sporadic sudden deaths in adult white-tailed deer occurred from November 1997 through August 1998 on an Iowa game farm. Three of the 4 deer necropsied had severe pulmonary edema, widespread mild lymphocytic vasculitis, and amphophilic intranuclear inclusion bodies in scattered endothelial cells in blood vessels in the lung and abdominal viscera. Immunohistochemistry with bovine adenovirus 5 antisera and transmission electron microscopy demonstrated adenoviral antigen and nucleocapsids, respectively, within endothelial cells. Adenovirus was isolated in cell culture from 1 of the affected deer. The isolate was neutralized by California black-tailed deer adenovirus antiserum. These findings indicate that adenovirus should be considered in the differential diagnosis of both black-tailed and white-tailed deer with pulmonary edema and/or hemorrhagic enteropathy.
Ribotyping and Restriction Endonuclease Analysis Reveal a Novel Clone of Bordetella Bronchiseptica in Seals
Authors: Register Karen B. , Sacco Randy E. , Foster Geoff
Published: May 2011

The goal of the present study was to characterize, by ribotyping and restriction endonuclease analysis (REA), 35 phocine Bordetella bronchiseptica isolates and to ascertain their relationship to one another and to isolates acquired from other host species. Thirty-four isolates were obtained in Scotland during a 10-year period encompassing the 1988 epizootic; the remaining isolate was obtained independently in Denmark. All phocine isolates had an identical Pvu II ribotype unique from the 18 ribotypes previously detected in strains from heterologous hosts. Alternative restriction enzymes, useful for subgrouping strains within Pvu II ribotypes, also failed to discriminate among isolates from seals. The exclusive occurrence of a single ribotype of B. bronchiseptica in a particular host species has not been previously observed. Similarly, REA based on either Hinfl or Dde I profiles did not reveal detectable polymorphisms, although unique patterns were readily distinguished among a limited number of isolates from other host species. This is the first report demonstrating the utility of REA using frequently cutting enzymes for discrimination of B. bronchiseptica strains. These data suggest that B. bronchiseptica-induced respiratory disease in seals along the Scottish shore may be due to the circulation of a single, unique clone.
Polymerase Chain Reaction for Definitive Identification of Yersinia Ruckeri
Authors: LeJeune Jeffrey T. , Rurangirwa Fred R.
Published: May 2011

Yersinia ruckeri causes enteric red mouth (ERM) disease in salmonids. Serologic identification of Y. ruckeri is hampered by cross-reactivity with other bacterial isolates of fish origin. Oligonucleotide primers incorporating Y. ruckeri unique sequences were designed to amplify a 409 bp fragment of Y. ruckeri 16S rDNA. The primers did not amplify other genetically related Yersinia or a wide variety of other aquatic or piscine bacteria. This assay provides a rapid, definitive identification of Y. ruckeri that is not subject to the variability inherent in serologic methods.
Adenovirus Hepatitis in a Boa Constrictor (Boa Constrictor)
Authors: Ramis A. , Fernández-Bellon H. , Majó N. , Martínez-Silvestre A. , Latimer K. , Campagnoli R.
Published: May 2011

A boa constrictor was submitted for postmortem evaluation. At necropsy, there were no substantial lesions except in the liver. Light microscopy revealed severe multifocal to coalescing coagulative necrotic hepatitis, with basophilic and eosinophilic intranuclear inclusions in hepatocytes within the necrotic foci. The histopathological findings suggested a viral hepatitis. An adenoviral infection was diagnosed by means of transmission electronic microscopy and in situ hybridization techniques.
An Indirect Enzyme-Linked Immunosorbent Assay for Diagnosis of American Canine Hepatozoonosis
Authors: Mathew John S. , Saliki Jeremiah T. , Ewing Sidney A. , Lehenbauer Terry W. , Panciera Roger J. , Malayer Jerry R. , Cummings Connie A. , Kocan A. Alan
Published: May 2011

American canine hepatozoonosis (ACH), caused by Hepatozoon americanum, is an emerging tick-borne disease of dogs. An indirect enzyme-linked immunosorbent assay (ELISA) that should facilitate diagnosis of infection and study of the epidemiology of ACH has been developed using H. americanum sporozoites as antigen. Efficacy of the new test as a diagnostic tool was compared with that of skeletal muscle biopsy, the current gold standard for confirming H. americanum infection. Results show that the test is sensitive (93%) and specific (96%) and that it is as reliable as histopathologic examination of skeletal muscle for detecting infection. The ELISA would be suitable as a routine laboratory test for diagnosis of ACH.
Validation of Synthetic Peptide Enzyme Immunoassays in Differentiating Two Subgroups of Ruminant Respiratory Syncytial Virus
Authors: Grubbs Steven T. , Kania Stephen A. , Potgieter L. N. D.
Published: May 2011

Subgroup-specific peptide-based enzyme immunoassays from each respective G-glycoprotein of the ovine and the bovine respiratory syncytial virus (RSV) were developed to detect RSV-specific IgG responses in cattle. Antigenic peptides from the respective G-glycoprotein were identified from the extracellular central hydrophobic region (amino acids 158–189) located between 2 mucin-rich regions. These antigenic peptides identified by epitope mapping from each G-glycoprotein were synthesized and used to develop the subgroup-specific enzyme immunoassays. The negative cutoff for each enzyme immunoassay was established as the mean optical density of indirect immunofluorescent antibody–negative bovine sera plus 3 SDs. The sensitivity (82.9%) and specificity (100%) of the bovine enzyme immunoassay and the specificity (95.8%) of the ovine enzyme immunoassay were determined by comparison with indirect immunofluorescence (used as the “gold standard”). The negative and positive predictive values were calculated for each assay. The presence of serum antibody to ovine RSV in cattle implies that this virus infects cattle and may contribute to the pathogenesis of bovine respiratory disease.
An Atypical Lymphoma of T-Cell Lineage in the Thorax of an Aged Cow
Authors: Step D. L. , Cummings C. A. , Streeter R. N. , Kirkpatrick J. G. , Campbell G. A.
Published: May 2011

An aged beef cow was presented for signs of thoracic disease. A complete clinical and diagnostic workup suggested neoplasia. Postmortem examination revealed a lymphoma of T-cell lineage confined solely to the thoracic cavity, predominantly in lung tissue. The diagnosis was based on light and electron microscopy, immunohistochemistry, and negative bovine leukemia virus and bovine immunodeficiency virus results.
Postweaning Multisystemic Wasting Syndrome Induced after Experimental Inoculation of Cesarean-Derived, Colostrum-Deprived Piglets with Type 2 Porcine Circovirus
Authors: Bolin Steven R. , Stoffregen William C. , Nayar Gopi P. S. , Hamel Andre L.
Published: May 2011

Cesarean-derived, colostrum-deprived pigs ( n = 23) were inoculated intranasally and subcutaneously with a low cell culture passage of type 2 porcine circovirus. In 11 pigs, a persistent fever that lasted 7–17 days began 12–15 days after inoculation with virus. Additional signs of disease in those 11 pigs included depression (11 of 11 pigs), palpable enlargement of inguinal, prefemoral, and popliteal lymph nodes (11 of 11), icterus (6 of 11), and hyperpnea (2 of 11). The remaining 12 pigs had fever that occurred intermittently for 2–4 days between days 12 and 20 postinoculation. Overt signs of disease in those pigs were limited to palpable enlargement of inguinal and popliteal lymph nodes (9 of 12 pigs). When compared with control pigs of similar age, the average daily rate of weight gain for all pigs inoculated with virus was less over a 2-week period that began 2 weeks post inoculation. At postmortem examination, lymph node enlargement was seen in 14 of 14 pigs euthanized between days 20 and 28 postinoculation. Lymph node enlargement was especially prominent in pigs that developed a persistent fever. Microscopic lesions noted in pigs that developed a persistent fever included cellular depletion in lymphoid tissues; hepatic cell necrosis; and lymphogranulomatous inflammation of lymph nodes, Peyer's patches of the intestine, liver, kidney, and heart. Virus was isolated with varying frequency from nasal, rectal, or tonsil swab specimens, buffy coat, serum, urine, and lung lavage fluid obtained antemortem or postmortem. Virus was isolated from or viral DNA was detected in a variety of tissues obtained postmortem up to 125 days postinoculation. Antibody against type 2 porcine circovirus usually was detected in serum between 15 and 20 days postinoculation; however, antibody against virus was not detected in serum from 4 pigs euthanized 20–24 days postinoculation. Direct contact with pigs inoculated with virus 42 days previously resulted in transmission of virus to 3 of 3 control pigs.
Isotype-Specific Antibody Responses of Cattle to Salmonella Dublin Lipopolysaccharide and Porin Following Salmonella Dublin Vaccination and Acute and Chronic Infection
Authors: House John K. , Smith Bradford P. , O'Connell Kevin , VanMetre David C.
Published: May 2011

Stimulation of different T-cell subsets during antigen presentation influences the antibody isotype response to an antigen. Salmonella infection and Salmonella bacterin vaccination are likely to stimulate different T-cell subtypes. The objective of this study was to determine whether there are differences in the isotype response of cattle to Salmonella antigens following Salmonella infection and Salmonella bacterin vaccination. Sera from Salmonella bacterin–vaccinated, experimentally infected, and chronically infected (carrier) adult cattle collected during previous studies was used to evaluate the IgG1, IgG2, and IgM isotype responses of cows to Salmonella serotype Dublin lipopolysaccharide (LPS) and porin. Following vaccination and experimental oral infection, IgG1 titers to LPS and porin rose more quickly and persisted longer than did IgG2 titers. In contrast to Salmonella infection, bacterin vaccination stimulated a weak response to Salmonella porin. Salmonella infection also induced a higher IgG2:IgG1 titer ratio to LPS than did bacterin vaccination. Chronic Salmonella infection induced the highest LPS and porin IgG2:IgG1 titer ratios and the highest correlation between LPS and porin titers. Response operating characteristic curves for each isotype-specific enzyme-linked immunosorbent assay (ELISA) were determined to evaluate the effect of isotype on the sensitivity and specificity of Salmonella ELISA serology for distinguishing sera of Salmonella carriers from those of vaccinated and acutely infected cows. IgG2 titers to LPS and porin provide a more specific indicator of chronic Salmonella infection status than do IgG1 titers to the same antigens with little to no loss in sensitivity.
Differential Detection of Bartonella Species and Strains in Cat Scratch Disease Diagnostics by Polymerase Chain Reaction Amplification of 16S Ribosomal RNA Gene
Authors: Tapp Ronald A. , Roy Alma F. , Corstvet Richard E. , Wilson Vincent L.
Published: May 2011

Cat scratch disease (CSD) has been difficult to diagnose in animals because of the protracted clinical course of infection and the quiescent phases when the microbial culprit lies dormant. The causative agent in CSD appears to be multiple species and strains of Bartonella. Using polymerase chain reaction (PCR) techniques for amplification of highly variable regions of the 16S ribosomal RNA (rRNA) gene sequence, a very sensitive species- and strain-specific assay for CSD-causing Bartonella species was developed. PCR primers were designed to specifically amplify the 16S rRNA gene of Bartonella species but not of other microbial pathogens. This initial PCR was multiplexed with a universal primer set, based on conserved sequence regions in the 16S rRNA gene, that provides a 162-bp fragment in all species tested. Subsequently, 3 distinct nested PCR primer sets enabled the individual amplification and specific detection of Bartonella henselae type I, B. henselae type II, and B. clarridgeae. Thus, this 2-step PCR procedure enabled the sensitive detection and identification of these species and the B. henselae genotype by exploiting minor sequences differences. Verification of these results were demonstrated with both sequencing and ligase chain reaction techniques. The diagnostic usefulness of this CSD test has been demonstrated by the analysis of specimens from control and infected cats. The diagnosis was confirmed and the specific B. henselae strain was correctly identified in peripheral blood specimens obtained from control and strain-specific CSD-infected cats. Such an accurate and sensitive diagnostic tool for the detection and identification of CSD causative agents should be a useful for the medical, veterinary, and scientific communities.
Septicemia Associated with Stenotrophomonas Maltophilia in a West African Dwarf Crocodile (Osteolaemus Tetraspis Subsp. Tetraspis)
Authors: Harris N. Beth , Rogers Douglas G.
Published: May 2011

A 17-year-old male captive West African dwarf crocodile ( Osteolaemus tetraspis subsp. tetraspis) died 1 month after fighting with a penmate. Abrasions were present on the head and mandible. Necropsy revealed a vegetative valvular lesion of the left atrioventricular valve, miliary foci of necrosis in the endocardium and myocardium, multiple duodenal and rectal ulcers, and serous atrophy of body fat. Stenotrophomonas maltophilia was isolated in pure culture from lung, liver, and kidney. Gram-negative bacilli were seen histologically in the valvular lesion and in foci of necrosis in the myocardium, liver, spleen, pancreas, kidney, and intestine. Septic thrombi in multiple tissues, arteritis, and pneumonia were additional histologic lesions. Findings indicated that the crocodile died from acute S. maltophilia septicemia, although the primary site of infection was not determined. Stenotrophomonas maltophilia is ubiquitous in the environment and is recognized as an important nosocomial pathogen in humans.
Multiplex Polymerase Chain Reaction for Distinguishing Taylorella Equigenitalis from Taylorella Equigenitalis-Like Organisms
Authors: Arata Amy B. , Cooke Cara L. , Jang Spencer S. , Hirsh Dwight C.
Published: May 2011

It is difficult to distinguish isolates of Taylorella equigenitalis, the cause of contagious equine metritis, from a T. equigenitalis-like organism isolated from asymptomatic donkeys and horses. Although T. equigenitalis is responsible for a severe, contagious disease of the reproductive tract of equids, the T. equigenitalis-like organism, although contagious, does not appear to produce disease. Because of the economic consequences of correctly distinguishing isolates of these 2 microorganisms, a polymerase chain reaction (PCR)-based assay was developed that will distinguish isolates of T. equigenitalis from the T. equigenitalis-like microorganism. The primers used in the PCR assay were designed to amplify unique regions of the gene encoding the 16S ribosomal RNA.
Partial Passive Protection with Two Monoclonal Antibodies and Frequency of Feeding of Hyperimmune Anti–Transmissible Gastroenteritis Virus (TGEV) Serum for Protection of Three-Day-Old Piglets from a TGEV Challenge Infection
Authors: Wesley Ronald D. , Woods Roger D.
Published: May 2011

Passive protection experiments were conducted to determine the frequency and amounts of hyperimmune antiserum needed to block a transmissible gastroenteritis virus (TGEV) challenge infection and to identify monoclonal antibodies that are partially protective against TGEV. Hyperimmune antiserum or monoclonal antibodies were added to milk at each feeding or at selected feedings when the amount of antiserum was reduced. Three-day-old piglets were challenged with virulent virus that had been preincubated with antiserum or monoclonal antibodies. The results indicated that supplementing antiserum every other day was not efficacious for protection. Supplementing even small quantities of hyperimmune antiserum (0.5 ml) at least once a day in most cases was sufficient for piglet survival but did not prevent morbidity. Increasing the amount (>2 ml) and providing antiserum 3 times/day completely blocked the TGEV challenge infection. Two monoclonal antibodies were discovered that also provided passive protection for baby pigs. One monoclonal antibody, 5G1, had a high neutralizing titer, and the other, 6C4, was more effective in neutralizing and binding to virulent TGEV than to attenuated TGEVs. Both of these monoclonal antibodies were partially effective as supplements in milk for passive protection. Furthermore, these monoclonal antibodies were useful for boosting the efficacy of TGEV-neutralizing colostrum, which by itself was ineffective. These results show that other antigenic sites, different from the 4-well characterized epitopes on the S glycoprotein of TGEV, also are important for passive protection.
Multiplex Polymerase Chain Reaction Assay for Identification of Enterotoxigenic Escherichia Coli Strains
Authors: Osek Jacek
Published: May 2011

A multiplex polymerase chain reaction (PCR) system was developed for identification of enterotoxigenic Escherichia coli (ETEC) strains and to differentiate them from other gram negative enteric bacteria. This test simultaneously amplifies heat-labile (LTI) and heat-stable (STI and STII) toxin sequences and the E. coli-specific universal stress protein ( uspA). The specificity of the method was validated by single PCR tests performed with the reference E. coli and non- E. coli strains and with bacteria isolated from pig feces. The multiplex PCR allowed the rapid and specific identification of enterotoxin-positive E. coli and may be used as a method for direct determination of ETEC and to differentiate them from other E. coli and gram-negative enteric isolates.
Complex Vertebral Malformation in a Holstein Calf: Report of a Case in the USA
Authors: Duncan Robert B. , Carrig Colin B. , Agerholm Jorgen S. , Bendixen Christian
Published: May 2011

Complex vertebral malformation (CVM), a familial syndrome of Holstein calves, has been reported in aborted fetuses and in prematurely born, stillborn, and neonatal calves. Affected calves have anomalies in the vertebral column, including hemivertebrae, fused and misshapen vertebrae and ribs, scoliosis, and vertebral synostosis. Concurrent low body weight, symmetrical arthrogryposis, and cardiac anomalies have been documented in affected calves. The syndrome was identified and characterized in Holstein cattle in Denmark; however, a global distribution of this genetic disorder is likely based on identification of common ancestral sires widely used for artificial insemination. This is the first documented case of CVM in a Holstein calf in the USA.
Detection and Confirmation of Reptilian Adenovirus Infection by in Situ Hybridization
Authors: Perkins Laura E. Leigh , Campagnoli Raymond P. , Harmon Barry G. , Gregory Christopher R. , Steffens W. L. , Latimer Ken , Clubb Susan , Crane Maria
Published: May 2011

Adenovirus infections are documented in at least 12 different species of reptiles. In contrast to their mammalian and avian counterparts reptilian adenoviruses are not well characterized as to their pathogenic potential and their ability to cause primary disease. In the diagnostic setting, fresh tissues are often not available for virus isolation, and the confirmation of reptilian adenovirus infections is dependent largely upon electron microscopy for the identification of intranuclear viral inclusions associated with histopathologic changes. The diagnosis of adenovirus infection in 2 different species of snake was confirmed by the application of DNA in situ hybridization. Using an aviadenovirus specific oligoprobe, adenoviral DNA was observed in the nuclei of hepatocytes, Kupffer cells, endothelial cells, and enterocytes. Electron microscopy of the liver confirmed the presence of intranuclear viral particles morphologically consistent with an adenovirus. DNA in situ hybridization on formalin-fixed tissues can serve as a suitable alternative to electron microscopy in the diagnosis of reptilian adenovirus infections. Both affected snakes had other concurrent diseases, suggesting that the adenovirus may not have been the primary pathogen.
In Vitro Detection of Shiga Toxin Using Porcine Alveolar Macrophages
Authors: Mengeling William L. , Vorwald Ann C. , Cornick Nancy A. , Lager Kelly M. , Moon Harley W.
Published: May 2011

Porcine alveolar macrophages were found to be highly susceptible to the cytolytic effects of a toxin (Shiga toxin [Stx]) produced by certain strains of Escherichia coli and sometimes associated with clinical disease in pigs and other animals. In comparison with the cells that are most commonly used for Stx detection and titration in vitro (namely, Vero cells), porcine alveolar macrophages appeared to be generally more sensitive and test results could be obtained in less time. Moreover, unlike Vero cells, porcine alveolar macrophages need not be continuously propagated to ensure immediate availability. They can simply be removed from a low-temperature repository, thawed, seeded, and shortly thereafter exposed to the sample in question. These characteristics suggest that porcine alveolar macrophages may be useful in developing a highly sensitive and timely diagnostic test for Stx.
Detection of Bovine Herpesvirus-1 in Peripheral Blood Mononuclear Cells Eight Months Postinfection
Authors: Wang Pu , Hurley David J. , Braun Lyle J. , Chase Christopher C. L.
Published: May 2011

Peripheral blood mononuclear cells (PBMCs) from 5 calves (3 controls and 2 vaccinates) used in a bovine herpesvirus 1 (BHV-1) vaccine study with a BHV-1 Cooper strain challenge were collected 6 months after challenge. The PBMCs from the control animals were positive by immunofluorescence for the BHV-1 glycoprotein D (gD) while the vaccinates were negative. The PBMC samples from 4 of the 5 animals were examined for BHV-1 DNA by polymerase chain reaction (PCR) and for gD immunofluorescence at 8 months after challenge. The BHV-1 DNA and viral antigen were detected in PBMC samples at 8 months postinfection, but no virus was isolated.
PCR Detection of the Cryptococcus Neoformans CAP59 Gene from a Biopsy Specimen from a Case of Feline Cryptococcosis
Authors: Kano Rui , Fujino Yasuhito , Takamoto Nobuko , Tsujimoto Hajime , Hasegawa Atsuhiko
Published: May 2011

A polymerase chain reaction assay was developed to detect Cryptococcus neoformans in biopsy samples. The assay detects the CAP59 gene of Cryptococcus neoformans and was used to substantiate cutaneous cryptococcosis in a 5-year-old cat submitted to the Veterinary Medicine Center at the University of Tokyo.
A Simple Thin-Layer Chromatographic Method for the Detection of Ergovaline in Leaf Sheaths of Tall Fescue (Festuca Arundinacea) Infected with Neotyphodium Coenophialum
Authors: Salvat Adriana E. , Godoy Héctor M.
Published: May 2011

A relatively simple and inexpensive thin-layer chromatographic (TLC) method is described for the detection and semiquantitative measurement of ergovaline in leaf sheaths of tall fescue ( Festuca arundinacea). Samples were finely ground and extracted with methanol. The extracts were filtered and the methanol was evaporated. The aqueous residue was extracted with hexane, followed by chloroform at pH 9. The chloroform extract was concentrated and further purified on a preparative silica gel TLC plate, developed with toluene/ethyl acetate/acetonitrile (50:10:40). The ergovaline band was scraped and eluted with methanol. The eluant was concentrated and an aliquot was applied to a silica gel TLC plate. The plate was developed successively with chloroform/acetone/acetic acid (90:10:5) and chloroform/ethanol (9:1). Ergovaline was visualized with p-dimethylaminobenzaldehyde and sulfuric acid. Semiquantitation of ergovaline was achieved by comparison with a known standard of ergotamine, which was shown to have the same Rf as ergovaline in this system. Spike recovery of ergotamine averaged 60%, with a limit of detection of 200 μg/kg of dry tall fescue leaf sheaths. The method was applied to 15 tall fescue samples with varying degrees of fungal infection, and ergovaline was identified in all contaminated samples with endophyte infection above 15%. Thin-layer chromatography may be also applicable for tall fescue seed, where the ergovaline content is usually higher and the amount of interfering pigments is much lower.
Detection of Circovirus Infection in Pigeons by in Situ Hybridization Using Cloned DNA Probes
Authors: Smyth Joan A. , Weston Jonathan , Moffett Deborah A. , Todd Daniel
Published: May 2011

Degenerate primers were designed based on known sequence information for the circoviruses psittacine beak and feather disease virus and porcine circovirus and applied by polymerase chain reaction (PCR) to known virus-infected bursa of Fabricius (BF) from a pigeon. A 548–bp DNA fragment was amplified and shown to be specific to a novel circovirus, named pigeon circovirus (PiCV), and was used to produce sensitive and specific probes for detection of circovirus DNA by in situ hybridization (ISH). Using ISH on BF from 107 pigeons submitted for necropsy, infection was detected in 89%, compared with a histologic detection rate of 66%. Using the ISH technique, infected cells were also found in liver, kidney, trachea, lung, brain, crop, intestine, spleen, bone marrow, and heart of some birds. Large quantities of DNA were present in some of these tissues, and in the absence of BF, liver in particular is identified as a potentially useful organ to examine for presence of PiCV. This high prevalence of infection in diseased birds is noteworthy, emphasizing the need for studies to determine the precise role of this virus as a disease-producing agent.
Serological Distinction of Bovine Salmonella Carriers from Vaccinated and Acutely Infected Cows
Authors: House John K. , Smith Bradford P. , Kamiya Darin
Published: May 2011

Identification of Salmonella carriers using lipopolysaccharide (LPS) ELISA serology in a Salmonella-infected herd requires distinction of chronically infected cattle from convalescent and vaccinated cows. Cows responding to Salmonella infection and vaccination produce titers to Salmonella LPS that overlap with the lower titers of some Salmonella carriers. The objective of this study was to determine if the LPS antigen specificity of the bovine humoral immune response to Salmonella LPS antigens differs following vaccination and acute and chronic Salmonella infection. The study focused on the nondiscriminatory area of Salmonella ELISA serology, specifically, peak-titered sera from Salmonella bacterin-vaccinated and experimentally infected cows and low-titered sera from Salmonella carriers. The LPS serogroup specificity of the IgG1 and IgG2 response following acute and chronic Salmonella serotype Dublin infection and Salmonella bacterin vaccination was evaluated using 5 Salmonella serogroup (B, D, E1, C3, and C1) LPS ELISA assays. IgG1 titers of carriers, vaccinated, and acutely infected cows were predominantly O antigen specific. Similarly, the IgG2 titers of acutely infected cows were also O antigen specific. In contrast, Salmonella carriers produced an IgG2 response to each of the heterologous LPS antigens (B, E1, C3, and C1) examined. The results of this study indicate that the bovine IgG1 isotype response to Salmonella LPS is serogroup specific. Conversely, production of IgG2 antibodies to core Salmonella LPS antigens shared across Salmonella serogroups is a feature of chronic Salmonella infections.
Development of Species-Specific PCR Techniques for the Detection of Tortoise Herpesvirus
Authors: Murakami Masaru , Matsuba Chikako , Une Yumi , Nomura Yasuo , Fujitani Hideo
Published: May 2011

Previously, a nested polymerase chain reaction (PCR) was employed with consensus degenerate primers targeting highly conserved motifs within herpesviral DNA polymerase genes to detect a newly described tortoise herpesvirus. However, nucleotide sequence information obtained from the final amplified fragment was restricted to a small region of 181 bp. In the present study, additional sequences flanking this segment were determined from a PCR product successfully amplified using a set of known degenerate primers, which covered a 692-bp region within the tortoise herpesviral DNA polymerase gene. Polymerase chain reaction primers for specific amplification of the tortoise herpesviral DNA were designed on the basis of these nucleotide sequences and successfully amplified tortoise herpesviral DNA from the tissues of tortoises that were well characterized histopathologically with herpesviral infection. The lower limit of detection was 1,000 herpesviral DNA equivalents in the presence of normal tortoise genomic DNA. Furthermore, a more sensitive and specific PCR technique for the identification of herpesviral infections in tortoises was developed employing a heminested form, which will enable the detection of latent infections or herpesvirus carriers in tortoises.
Mycobacterium Marinum Dermatitis and Panniculitis with Chronic Pleuritis in a Captive White Whale (Delphinapterus Leucas) with Aortic Rupture
Authors: Bowenkamp Kathryn E. , Frasca Salvatore , Draghi Andrew , Tsongalis Gregory J. , Koerting Claudia , Hinckley Lynn , de Guise Sylvain , Montali Richard J. , Goertz Caroline E. C. , St. Aubin David J. , Dunn J. Lawrence
Published: May 2011

A 16-year-old female white whale, Delphinapterus leucas, died after nearly 18 months of chronic lymphopenia and pyogranulomatous dermatitis. Necropsy revealed rupture of the aorta with hemorrhage into the cranial mediastinum and between fascial planes of the ventral neck musculature. Multiple foci of ulcerative dermatitis and panniculitis were present across the thorax and abdomen and surrounded the genital folds. In addition, there was a chronic proliferative pleuritis with over 20 liters of histiocytic exudate in the thoracic cavity. Acid-fast bacteria consistent with Mycobacterium sp. were identified in sections of skin lesions and in cytospins of pleural exudate. Cultures of pleura and 1 skin lesion collected at necropsy yielded sparse growth of an acid-fast bacillus with colony characteristics and morphology consistent with Mycobacterium marinum. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis confirmed the presence of M. marinum DNA in samples of skin. This is the first documented occurrence of mycobacteriosis in a white whale and is a unique presentation of mycobacterial dermatitis and panniculitis with chronic pleuritis in a cetacean. The improved PCR-RFLP protocol utilized in this case unifies techniques from several protocols to differentiate between species of Nocardia and rapidly growing mycobacteria clinically relevant to aquatic animals.