bluuebunny/crossref_metadata_2025 huggingface dataset:
Involvement of Canine Oral Papillomavirus in Generalized Oral and Cutaneous Verrucosis in a Chinese Shar Pei Dog
Authors: Sundberg J. P. , Smith E. K. , Herron A. J. , Jenson A. B. , Burk R. D. , Van Ranst M.
Published: January 2010

Severe papillomatosis developed in the oral cavity and spread throughout the haired skin of the trunk and limbs of an 8-month-old female Chinese Shar Pei dog. The dog had received corticosteroids prior to referral, which was associated with the onset of demodecosis and papillomatosis. Papillomavirus structural antigens were detected in biopsies by immunohistochemistry using a panel of monoclonal and polyclonal antibodies. An 8.2-kilobase papillomavirus-specific DNA molecule was detected in the cutaneous lesions by high stringency Southern blot hybridization using a cloned canine oral papillomavirus DNA probe. Restriction enzyme analysis revealed that the virus in the cutaneous lesions was identical to the canine oral papillomavirus. Discontinuation of the steroids combined with the use of a mitocide, antibiotics, and an autogenous vaccine resolved the demodecosis and papillomatosis. This case report suggests that corticosteroid-induced immunosuppression can expand the tissue tropism of papillomaviruses.
Proteinuria Associated with Glomerulosclerosis and Glomerular Collagen Formation in Three Newfoundland Dog Littermates
Authors: Koeman J. P. , Biewenga W. J. , Gruys E.
Published: January 2010

Three dogs out of a litter of eight Newfoundland dogs developed a progressive fatal glomerulopathy. The affected dogs were a 2-month-old male, a 2.5-month-old female, and a 1-year-old male. The disease in all three animals was characterized by growth retardation, anorexia, proteinuria (14–16 g/liter), hypoalbuminemia (15–21 g/liter, elevated plasma urea (13–28 mmol/liter), and creatinine (83–296 μmol/liter) concentrations. Because of a bad prognosis the dogs were euthanatized. On postmortem examination, the animals had enlarged, slightly pale kidneys, which revealed glomerulosclerosis and glomerulofibrosis on histologic and electron microscopic examination. The lesions consisted of subendothelial and mesangial collagen fibrils and an increase of mesangial matrix. The fibrosis may result from endothelial or mesangial collagen formation as the manifestation of a metabolic disease.
Absence of Glomerulonephritis in Guinea Pigs Deficient in the Fourth Component of Complement
Authors: Foltz C. J. , Cork L. C. , Winkelstein J. A.
Published: January 2010

Genetically determined deficiencies of the early components of the classical complement pathway (CI, C4, C2) or of the third component of complement (C3) in both human beings and experimental animals are known to be associated with renal disease, including glomerulonephritis. The current study was performed to examine the C4–deficient (C4D) guinea pig for the presence of renal disease. Eighteen C4D animals and 17 control animals (Crl:Hartlcy) (divided by sex into four age categories) were examined. Light microscopic examination revealed no differences in mesangium, glomerular cellularity, thickness of capillary loops, or presence of epithelial crescents in the kidneys of C4D guinea pigs as compared with control animals. Electron microscopic examination did not reveal glomerular or tubular immune complex deposits in either C4D or control animals. C4D guinea pigs apparently do not demonstrate glomerulonephritis.
Flow Cytometric DNA Ploidy Analysis in Canine Transitional Cell Carcinoma of Urinary Bladders
Authors: Clemo F. A. S. , DeNicola D. B. , Carlton W. W. , Morrison W. B. , Walker E.
Published: January 2010

Flow cytometric analysis of DNA ploidy was performed on 51 formalin-fixed, paraffin-embedded samples of canine transitional cell carcinoma of the urinary bladder. The DNA ploidy data obtained were compared to several clinicopathologic features. In addition, the DNA ploidy of 15 hyperplastic/inflamed and 8 normal canine urinary bladders was measured. Forty-three of the 51 neoplastic samples had interpretable DNA histograms. DNA aneuploidy was found in 34/43 (79%) of the transitional cell carcinoma samples. Of the 34 aneuploid neoplasms, 16 (47%) were hyperdiploid, 17 (50%) were tetraploid, and 1 (3%) was hypertetra-ploid. No significant correlation was found between the presence of DNA aneuploidy and the growth pattern. histologic grade, clinical stage, or individual morphologic features of this neoplasm. Additionally, the DNA ploidy was not related to the sex, age, or survival time of dogs with transitional cell carcinoma. All of the normal and all but one of the hyperplastic/inflamed urinary bladders were diploid. The results from this study demonstrated that DNA ploidy can be measured from paraffin-embedded canine samples by flow cytometry, a majority of the canine transitional cell carcinomas were aneuploid, and a significant correlation did not exist between the DNA ploidy and specific clinicopathologic features of this neoplasm.
Infected Cell Types in Ovine Lung Following Exposure to Bovine Respiratory Syncytial Virus
Authors: Meehan J. T. , Cutlip R. C. , Lehmkuhl H. D. , Kluge J. P. , Ackermann M. R.
Published: January 2010

Sixteen adult sheep (ten females, six males obtained from a closed flock at National Animal Disease Center, Ames, IA) were experimentally infected with bovine respiratory syncytial virus strain 375 (BRSV), and lung tissues were stained for viral antigen. Two infected sheep were euthanatized at each of the following post-inoculation times: 12, 24, 36, 48, 72, 96, 144, and 192 hours. Lung, nasal turbinates, trachea, right cranial bronchial and mediastinal lymph nodes, liver, and spleen were collected for histologic evaluation. An indirect immunoperoxidase technique was performed on routine paraffin-embedded sections of lung tissue, trachea, turbinates, and bronchial and mediastinal lymph nodes to determine the location of the BRSV antigen. For lung tissue from each sheep 400 light microscopic fields at 160x magnification were examined for staining for BRSV antigen. Lung tissue was also collected for virus and bacterial isolation. Daily serum samples were taken for determination of anti-BRSV titers. Severe respiratory disease was not produced in any sheep. Bovine respiratory syncytial virus was isolated from lung tissue collected from all sheep up through 144 hours postinoculation. At 12 hours post-inoculation (case No. 2) respiratory syncytial virus antigen was detected in bronchiolar epithelium and a mononuclear cell within an alveolar space. Lung tissue from the sheep necropsied between 24 and 144 hours postinoculattion (case Nos. 3–14) contained BRSV antigen in bronchiolar epithelium, type I pneumocytes, type II pneumocytes. alveolar macrophages, and mononuclear cells within alveolar spaces. Macrophages staining for viral antigen were rare. Bronchiolar and type I epithelial cells comprised the majority of infected cells. In a separate experiment, lung slices inoculated in vitro with either BRSV or ovine adenovirus did not stain for the respective antigens. Slices inoculated with parainfluenzavirus-3 did stain for that viral antigen.
Comparative Pathology of a Chicken-origin and Two Duck-origin Influenza Virus Isolates in Chickens: The Effect of Route of Inoculation
Authors: Swayne D. E. , Slemons R. D.
Published: January 2010

Forty-nine 5–week-old chickens were inoculated by the intravenous (IV), intratracheal (IT), or intranasal (IN) routes with either a chicken-origin or one of two duck-origin type A influenza virus isolates. Twelve control chickens were inoculated with sterile chorioallantoic fluid. For all viruses, IV inoculation produced predominate lesions of renal tubule necrosis (nephrosis) and nephritis, and influenza virus nuclcoprotein was localized in nuclei and cytoplasm of necrotic renal tubule epithelium. Chickens inoculated by the IT route, and to a lesser extent the IN route, had mild to severe tracheitis, bronchitis, and ventromedial pneumonia associated with secondary bronchi but lacked renal tubule necrosis and nephritis. These data indicate low-virulence avian-origin influenza viruses were nephrotropic during simulated systemic infection (IV inoculation) and pneumotropic during simulated local infection (IT and IN inoculation). Gross and histologic kidney lesions produced by IV inoculation of the chicken-origin influenza virus were similar to changes reported in outbreaks of low-virulence influenza virus in laying chickens.
Light Microscopic, Ultrastructural, and Immunohistochemical Examinations of Two Calcifying Epithelial Odontogenic Tumors (CEOT) in a Dog and a Cat
Authors: Breuer W. , Geisel O. , Linke R. P. , Hermanns W.
Published: January 2010

Two calcifying epithelial odontogenic tumors from a 10-year-old cross-breed dog and a 9-year-old cat were characterized histologically and electron microscopically. Important characteristics were plexiform to squamous epithelial structure, amyloid in the tumor tissue with partial calcification (Liesegang's rings), and a low rate of mitosis. The amyloid found in the tumors was subjected to immunohistochemical examination, using a series of antibodies (anti-AA, -AL, -AF, -ASc1, -AB, -cytokeratin, -vimentin, -desmin, -laminin). The exclusive reaction to anti-cytokeratin and anti-laminin suggests that the amyloid is of epithelial origin.
Familial Nephropathy in Bernese Mountain Dogs
Authors: Minkus G. , Breuer W. , Wanke R. , Reusch C. , Leuterer G. , Brem G. , Hermanns W.
Published: January 2010

Between January 1988 and March 1992 nephropathies were frequently diagnosed in Bernese Mountain Dogs. During this period, 20 animals (16 females, four males), ages 2–5 years (average age at time of diagnosis = 3.3 years) presented with clinically renal insufficiency. Morphologic diagnosis of the renal lesions was identical in all cases, i.e., membranoproliferative glomerulonephritis (MPGN) with concomitant interstitial nephritis. Deposits of immunoglobulin-M (IgM) and of the third complement component were regularly demonstrated immunohistochemically in the glomeruli; deposits of immunoglobulin-A (IgA) and immunoglobulin-G (IgG) were found only in isolated cases. Reduplication of glomerular basement membranes, mesangial interposition, and subendothelial deposits of the immunocomplex type were also detected by electron microscopy. A pedigree analysis indicated that the MPGN in these 20 Bernese Mountain Dogs of approximately the same age was of hereditary genesis. Thus, MPGN should be allocated to the group of familial nephropathies. Serologically, high IgG titers against Borrelia burgdorferi were found in 17 dogs. These findings are discussed in relation to familial nephropathies in humans.
Multiple Osteomas in Mice
Authors: Höger H. , Gialamas J. , Jelinek F.
Published: January 2010

Osteomas (dense compact neoplasms of mature bone tissue) are rare in nearly all strains and stocks of mice. Of 224 Him: OF1 mice maintained until natural death or until terminally ill. 116 (51.8%) had one or more osteomas. Osteomas had a predilection for the skull and the larger bones of the limbs. Plasma alkaline phosphatase concentrations were elevated significantly in osteoma-bearing mice (446 ± 153 U/liter versus 206 ± 65 U/liter in age-matched controls without osteomas). Only very large osteomas resulted in clinical signs, and longevity was not shortened. Histologic examination showed clearly separated dense bony tissue irregularly arranged and forming a mosaic pattern, with distinct cement lines and medullary spaces filled with fibroreticular connective tissue. Electron microscopic examination revealed virus-like structures in osteoblasts, osteocytes, and fibroblasts and in the place of remnants of necrotic cells.
Feline Mucopolysaccharidosis VII Due to β-glucuronidase Deficiency
Authors: Gitzelmann R. , Bosshard N. U. , Superti-Furga A. , Spycher M. A. , Briner J. , Wiesmann U. , Lutz H. , Litschi B.
Published: January 2010

A male cat 12–14 weeks old had walking difficulties and an enlarged abdomen. Facial dysmorphism, plump paws, corneal clouding, granulation of neutrophils, vacuolated lymphocytes, and a positive urine test for sulfated glycosaminoglycans suggested mucopolysaccharidosis. Cultured fibroblasts incorporated 35SO4 into mucopolysaccharides more actively than did fibroblasts of a feline control, and degradation was far inferior. Activity of β-glucuronidase was absent in leukocytes and markedly reduced in fibroblasts, thus establishing the diagnosis of mucopolysaccharidosis VII, a disorder previously described in humans, dogs, and mice. Light microscopic examination revealed foam cells in virtually all organs examined, and electron microscopic examination showed pancytic storage of floccular material characteristic of mucopolysaccharides. Stored sphingolipids in the form of zebra bodies were seen in ganglion cells of the central nervous system and in smooth muscle cells of blood vessels. This case represents another animal model of mucopolysaccharidosis VII with the full disease characteristics known in human patients.
Immunohistochemical Detection of Melanoma-associated Antigens on Formalin-fixed, Paraffin-embedded Canine Tumors
Authors: Berrington A. J. , Jimbow K. , Haines D. M.
Published: January 2010

Seven monoclonal antibodies (Moabs), recognizing melanoma-associated antigens in human tissues, were evaluated for their ability to immunohistochemically stain formalin-fixed, paraffin-embedded canine melanomas. Only 2 Moabs, designated human melanosome-specific antigen (HMSA)-1 and HMSA-5, stained routinely processed canine melanomas, staining 21/35 (60%) and 24/35 (69%), respectively. Twenty-nine of 35 (83%) melanomas tested were stained if results of the 2 Moabs were combined. Monoclonal antibody HMSA-1 also stained neoplastic cells of 10/35 (29%) tumors of nonmelanocytic origin and some neurons and salivary gland epithelial cells in normal canine tissues. However, Moab HMSA-1 staining in the nonmelanocytic tumors, consisting of small, discrete periodic acid-Schiff-positivc cytoplasmic droplets, was readily distinguishable from the diffusely granular, cytoplasmic staining of melanocytic tumors. In addition to melanomas, Moab HMSA-5 stained melanocytes and some melanin-containing tumor cells of a pigmented basal cell tumor and melanocytes in normal canine skin. Monoclonal antibodies HMSA-1 and HMSA-5 immunohistochemically identified the majority of canine melanomas, had limited and distinguishable staining in normal tissues and nonmelanocytic tumors, and therefore may be a useful adjunct for the diagnosis of canine melanoma in formalin-fixed, paraffin-embedded tissues.
Cardiovascular Lesions in Bovine Marfan Syndrome
Authors: Potter K. A. , Besser T. E.
Published: January 2010

Bovine Marfan syndrome is a genetic disease with many of the clinical and pathologic manifestations of human Marfan syndrome. Major manifestations include ectopia lentis and aortic dilatation, aneurysm, and rupture. Affected cattle have a defect in fibrillin metabolism similar to that in human patients. Ten cattle were followed and their disease progression and lesions documented. Ages ranged from a term fetus (No. 9) to a 4-year-old cow (No. 4); three animals were male (Nos. 1–3) and seven were female (Nos. 4–10). Of eight animals (80%) that died or were euthanatized (Nos. 1–3, 5–9), six (75%) had severe cardiovascular lesions identified at necropsy. Gross cardiovascular lesions of bovine Marfan syndrome included cardiac tamponade secondary to aortic rupture (animal Nos. 3, 6, 8), dissecting aneurysms of the aorta and pulmonary artery (animal No. 5), and intrauterine cardiac tamponade secondary to rupture of the pulmonary artery (animal No. 9). Microscopically, Verhoeff Van Gieson-stained sections of aorta contained severe fragmentation of the elastic laminae in the aortic media, but the cystic medial necrosis seen in human Marfan aortae was not identified, even in the chronic aortic dissection. Ultrastructurally, affected aortic tissue was characterized by thin, dark elastic fibers with abundant, tangled microfibrils on the periphery, Swirls of collagen fibers and bundles of hypertrophic smooth muscle cells replaced damaged elastic laminae. Gross and microscopic cardiovascular lesions in bovine Marfan syndrome are similar to those in human Marfan syndrome. Bovine Marfan syndrome is a valuable model for investigation of molecular pathogenesis and treatment of human Marfan syndrome.
Diagnostic and Prognostic Importance of Chromosomal Aberrations Identified in 61 Dogs with Lymphosarcoma
Authors: Hahn K. A. , Richardson R. C. , Hahn E. A. , Chrisman C. L.
Published: January 2010

To determine the diagnostic and/or prognostic importance of chromosomal aberrations identified in dogs with malignant (non-Hodgkin's) lymphoma, clinical stages for 61 dogs with lymphosarcoma were determined, the lymph node(s) were histopathologically graded, and the malignant tissue lymphocytes were karyotyped. The results from life table survival curve analysis demonstrated that first remission length and survival time were significantly longer in 15 of 61 (25%) dogs that had a trisomy of chromosome 13 as the primary chromosomal aberration than in those dogs (46/61, 75%) with other primary chromosomal aberrations ( P < 0.05). Sex, age, weight, histopathologic subtype and grade, World Health Organization (WHO) clinical stage, WHO and modified Karnofsky performance status, chromosomal modal number, and treatment protocol were of no prognostic importance in predicting first remission length or survival time ( P > 0.05). Multivariate analysis did not identify a significant correlation between the prognostic groups or within the various prognostic subsets ( P > 0.05). The pathogenesis of canine and human non-Hodgkin's lymphoma, as observed cytogenetically, differs.
Detection of Activated Feline Platelets in Platelet-rich Plasma by Use of Fluorescein-labeled Antibodies and Flow Cytometry
Authors: Welles E. G. , Bourne C. , Tyler J. W. , Boudreaux M. K.
Published: January 2010

Platelets contribute to prethrombotic or thrombotic states; however, accepted evaluation methods (i.e., in vitro testing by use of an aggregometer) of platelet function in cats can be difficult because of the large volume of blood required from which platelets are isolated and the potential for platelet activation due to difficult venipunctures in sometimes uncooperative or excited animals. The activation problem also contributes to errors in platelet counts. Platelets from four domestic short haired cats (two males, two females, 2–3 years old) minimally restrained without sedation or anesthesia were evaluated. Blood (5 ml) was collected by jugular venipuncture directly into syringes containing 3.8% trisodium citrate (nine parts blood to one part anticoagulant) plus prostaglandin E1 (3 μM; 0.25, 0.5, 1, or 2 μl/500 μl citrate) or 3.8% trisodium citrate alone. Prostaglandin E1, which is a stable metabolite of arachidonic acid with platelet inhibitory properties similar to those of prostaglandin I2 was added to the anticoagulant to prevent activation of platelets during the collection process. Feline platelets exposed to prostaglandin E1 became immediately and persistently nonreactive to agonists, which negated their use in functional studies (aggregation, 14C-serotonin release, binding of fluorescein-conjugated antifibrinogen) but improved platelet counting accuracy. Detection of in vivo activation of platelets in prethrombotic and thrombotic states in humans has been done by identification of activation-dependent molecules on platelet surfaces by use of specific antibody recognition and detection by flow cytometric analysis. Many activation-dependent platelet surface receptor changes are species specific; however, fibrinogen appears to be conserved across species. Results of flow cytometric analysis by forward and side angle light scatter easily identified feline platelets in 10 μl of platelet-rich plasma but were less definitive in identification of platelets in whole blood. Feline platelets in platelet-rich plasma activated by ADP (5 or 50 μM), which allowed binding of fibrinogen to exposed fibrinogen receptors, were labeled with fluorescein-conjugated anti-human fibrinogen. Labeled platelets were identified as a separate population by flow cytometric staining pattern and fluorescence intensity from nonlabeled platelets or nonspecifically-labeled resting platelets. This technique should allow detection of activated feline platelets from extremely small quantities of blood, which could facilitate repetitive and serial platelet testing in diseases thought to be accompanied, caused, or augmented by thrombotic conditions.
Presence of African Horse Sickness Virus in Equine Tissues, as Determined by in situ Hybridization
Authors: Brown C. C. , Meyer R. F. , Grubman M. J.
Published: January 2010

In a retrospective study, a negative-sense digoxigenin-labeled RNA probe, corresponding to the gene encoding nonstructural protein-1 of African horse sickness virus (AHSV) serotype 4, was applied to formalin-fixed, paraffin-embedded tissue taken from horses in the terminal stages of infection with AHSV, Fifteen infected ponies and one noninfected control were studied. Ponies exhibited a range of clinical signs and lesions. Thirteen ponies were infected with serotype 4, one with serotype 1, and one with serotype 2. Ponies were monitored clinically and euthanatized when severely clinically ill. The following tissues were available for study by in situ hybridization and histopathology: lung, heart, spleen, neck muscle, and supraorbital fat. Histologically, the most striking changes were pulmonary edema and, in some, acute myocardial necrosis. In situ hybridization revealed virus distributed widely in sections of lung and heart examined, with relatively less in spleen, neck muscle, or supraorbital fat. Virus was localized to target cells with morphologic features compatible with endothelium in all organs except spleen, where it was found in both endotheliumlike cells and large mononuclear cells.
Morphologic Changes in the Nasal Cavity Associated with Sialodacryoadenitis Virus Infection in the Wistar Rat
Authors: Bihun C. G. D. , Percy D. H.
Published: January 2010

A sequential study of lesions of the nasal cavity associated with sialodacryoadenitis virus (SDAV) infection was made in the laboratory rat. Wistar rats were intranasally inoculated with approximately 103TCID50of the Coronavirus SDAV. Transverse sections of four regions of the nasal cavity from inoculated and control animals were examined by light microscopy and immunohistochemistry at 2, 4, 6, 8, 10, and 14 days postinoculation (PI). Lesions were observed in the following regions of the upper respiratory tract: respiratory epithelium, transitional epithelium, olfactory epithelium, nasolacrimal duct, vomeronasal organ, and the submucosal glands of the nasal passages. In general, in structures lined by ciliated epithelial cells, there was focal to segmental necrosis with exfoliation of affected cells and polymorphonuclear cell infiltration during the acute stages, progressing to squamous metaplasia during the reparative stages. Repair in these regions was essentially complete by 14 days PI. In the olfactory epithelium and the vomeronasal organ, there was interstitial edema with necrosis and exfoliation of epithelial cells and minimal to moderate inflammatory cell response during the acute stages. Residual reparative lesions were still evident in the olfactory epithelium, the columnar epithelium and neuroepithelium of the vomeronasal organ, and the nasolacrimal duct at 14 days PI. Viral antigen was demonstrated by immunohistochemistry in all regions during the acute stages of the disease, with the exception of the vomeronasal organ. In view of these findings, infections of the respiratory tract with viruses such as SDAV could have significant effects on functions such as olfaction and chemoreception for ≥ 2 weeks postexposure in this species.
Occurrence of Keratoconjunctivitis Apparently Caused by Mycoplasma gallisepticum in Layer Chickens
Authors: Nunoya T. , Yagihashi T. , Tajima M. , Nagasawa Y.
Published: January 2010

Natural cases of keratoconjunctivitis, apparently caused by Mycoplasma gallisepticum (MG), in layer chickens are described. The disease occurred in a commercial flock consisting of 36,000 pullets (Babcock), first appearing around 30 days of age. Clinically, affected chickens showed unilateral or bilateral swelling of the facial skin and the eyelids, increased lacrimation, congestion of conjunctival vessels, and respiratory rales. Some of the severely affected chickens closed their eyes. The morbidity reached 27.8%, and it was estimated that approximately 10% died from reduced feed intake due to impaired vision. Ten 70-day-old chickens with clinical diseases were examined for lesions. There was acute to subacute keratoconjunctivitis in all of the chickens, and some exhibited laryngitis. Adherence of mycoplasma organisms to epithelial cells of the conjunctiva, cornea, and larynx was frequently observed. These organisms had an ultrastructure characteristic of MG and showed a positive reaction with rabbit polyclonal antibodies against the S6 strain of MG by immunohistochemical analysis. MG was isolated from tissue homogenates of the eyelids and tracheas of the affected chickens. Many of the chickens had atrophic bursae, and infectious bursal disease virus antigens were detected in necrotic bursal follicles by immunostaining. Therefore, immunosuppression due to infectious bursal disease was implicated in the pathogenesis of keratoconjunctivitis in the present cases.
Ultrastructural and Histomorphometric Evaluations of Gallium Nitrate on Bone in Nude Mice Bearing a Canine Adenocarcinoma (CAC-8) Model of Humoral Hypercalcemia of Malignancy
Authors: Okada H. , Merryman J. I. , Capen C. C. , Rosol T. J.
Published: January 2010

Gallium is a group IIIa transition metal that lowers serum calcium by an unknown mechanism and has been utilized in the treatment of cancer-associated hypercalcemia. The purpose of this study was to histomorphometrically investigate the ultrastructural effects of gallium nitrate on osteoclasts and osteoblasts in trabecular bone of normal nude mice and nude mice with humoral hypercalcemia of malignancy. Two groups of normal nude mice ( n = 7 and n = 8, respectively) and two groups of hypercalcemic nude mice ( n = 9) bearing a serially transplantable canine adenocarcinoma (CAC-8) were treated with vehicle or gallium nitrate. Osteoclasts were hypertrophied in vehicle-treated tumor-bearing nude mice as compared with vehicle-treated nontumor-bearing nude mice. Osteoclasts from tumor-bearing nude mice treated with gallium nitrate were significantly decreased in size and had fewer intracytoplasmic vesicles as compared with osteoclasts from vehicle-treated tumor-bearing nude mice. Degenerate osteoclasts, characterized by pyknotic nuclei and increased cytoplasmic vacuolation, were observed in both groups of gallium-treated nude mice. Osteoblasts from vehicle-treated tumor-bearing nude mice were hypertrophied and had extensive lamellar arrays of rough endoplasmic reticulum as compared with osteoblasts from vehicle-treated nontumor-bearing nude mice. Osteoblasts in gallium-treated nude mice (tumor-bearing and nontumor-bearing) were small and flattened with poorly developed cytoplasmic organelles. This investigation demonstrated that osteoclasts and osteoblasts in nude mice treated with gallium nitrate had ultrastructural evidence of decreased metabolic and functional activity. The results suggest that gallium nitrate lowers serum calcium by inhibiting osteoclastic bone resorption.
Local Production of Tumor Necrosis Factor-α in Corynebacterial Pulmonary Lesions in Sheep
Authors: Ellis J. A. , Campos M. , Snyder M. , Chelak B. , Haines D. M.
Published: January 2010

Corynebacterium pseudotuberculosis infection is a common cause of pyogranulomas in ovine lungs and often occurs as a dual infection with lentiviruses. This coinfection usually leads to the development of chronic pneumonia and cachexia that is similar to the clinical syndrome seen in human beings with AIDS-related pneumonias. Recent in vitro studies indicate that monokines such as tumor necrosis factor-α (TNFα) are induced by C. pseudotuberculosis, suggesting that TNFα is involved in the pathogenesis of corynebacterial lesions in vivo. To substantiate in vitro observations concerning bacterial induction of TNFα in ovine pulmonary macrophages, immunohistochemical labeling techniques were used in combination with in situ hybridization to identify TNF-producing cells in corynebacterial lesion sites in vivo. TNFα message and translation product were found in macrophages comprising pyogranulomas that were induced by naturally acquired and experimental pulmonary C. pseudotuberculosis infections.
A Comparative Study of the Histopathologic Features of Bovine Tuberculosis in Cattle, Fallow Deer (Dama dama), Sika Deer (Cervus nippon), and Red Deer and Elk (Cervus elaphus)
Authors: Rhyan J. C. , Saari D. A.
Published: January 2010

Sections of tuberculous lesions from 23 elk ( Cervus elaphus nelsoni) and red deer ( Cervus elaphus elaphus), 12 fallow deer ( Dama dama), 10 sika deer ( Cervus nippon), and 30 cattle were examined and compared. Lesions were scored for caseous necrosis, mineralization, neutrophils, macrophages, giant cells, and acid-fast bacilli. Some differences in lesion morphology between the species were noted. Elk/red deer lesions had marked variation and often differed from bovine lesions in several characteristics; elk/red deer lesions usually had scattered peripheral mineralization rather than central mineralization and contained more neutrophils and fewer giant cells than did bovine lesions. Fallow deer lesions contained more giant cells but were otherwise indistinguishable from elk lesions. Sika deer lesions had more giant cells and fewer neutrophils than did lesions from cattle or other cervid species. Sika deer giant cells were larger and contained more nuclei than did giant cells in the other species.
Renal Glomerular Fibrosis in Two Pigs
Authors: Shirota K. , Masaki T. , Kitada H. , Yanagi M. , Ikeda Y. , Une Y. , Nokmura Y. , Jothy S.
Published: January 2010

Massive deposition of collagen fibrils was found in the renal glomeruli of two pigs. The affected pigs were a 6-month-old female hybrid pig with systemic mycobacteriosis and a retired sow showing azotemia. Microscopically, various amounts of a homogeneous eosinophilic substance were deposited within the expanded mesangium of almost all renal glomeruli in both cases. The deposit was also seen occasionally on the glomerular capillary walls in the sow. Capillary lumens were occluded in the glomerular segments with heavy deposition. Obsolescent glomeruli were often surrounded by lymphoid cells. The deposited substance was Congo red negative and stained deep blue with Heidenhain's azan staining. Immunohistochemical evaluation revealed that the major component of the homogeneous substance was type-Ill collagen. Electron microscopic examination showed that the deposits were masses of fibrils of variable length, width, and shape. The fibrils had transverse bands with a periodicity of approximately 60 nm, consistent with collagen fibrils. These glomerular changes were identical to those of collagenofibrotic glomerulonephropathy that has recently been described as a new glomerular disease in humans. The glomerular lesion may be the result of unusual production of type-III collagen by mesangial cells.
Elaeophorosis in Red Deer Caused byElaeophora elaphi: Lesions of Natural Disease
Authors: Carrasco L. , Fierro Y. , Sánchez-Castillejo J. M. , Bautista M. J. , Gómez-Vllamandos J. C. , Sierra M. A.
Published: January 2010

This is the first study of the lesions produced by Elaeophora elaphi, a filarial worm first reported in 1986. Only the morphologic characteristics of this species have been described previously. Samples for the study were taken from 74 red deer ( Cervus elaphus), 1-10 years old, slaughtered in central Spain. Grossly, filariae were observed in the transverse, left, or right branches of the portal vein of 18 animals. In addition to filariae, whitish thrombi were found in the right and left branches of the portal vein in nine animals. Thrombi of similar characteristics were also observed in 12 deer in which no adult filariae were found. Microscopic changes were observed only in the right and left branches of the portal vein and consisted of thickening of the intima with surface formation of villi. Stroma of villi contained a moderate to intense infiltrate composed mainly of eosinophils and mononuclear cells. The tunica media of affected vessels was uniformly hypertrophic and in some cases presented a moderate inflammatory infiltrate similar to that observed in the stroma of villi. Thrombi consisted of a central area containing calcified filariae or cuticle debris, surrounded by giant cells and numerous lymphoid cells forming clearly defined nodules resembling lymphoid follicles. Thrombi were externally surrounded by a strip of connective tissue and beyond by endothelial cells that were occasionally continuous with the tunica intima. Other lesions probably associated with the infestation were multifocal granulomas in the periportal areas, lymphoid tissue formations in the hepatic parenchyma, and an interstitial nephritis.
Morphologic Changes of the Ascending Colon during Experimental Ischemia and Reperfusion in Ponies
Authors: Darien B. J. , Stone W. C. , Dubielzig R. R. , Clayton M. K.
Published: January 2010

The morphologic changes following ascending colon volvulus result from the interaction of inflammatory and coagulation mediators. The objective of this study was to establish a quantifiable histopathologic scoring system to evaluate the serial pathomorphologic changes during ischemia and reperfusion. Such a scoring system could then be applied to subsequent studies designed to attenuate bowel lesions by regulating activity of individual mediators. Ten normal, healthy adult ponies were randomly divided into two equal groups. Following anesthesia and a 30-minute stabilization period, the colon of ponies in the experimental group was twisted 720°: for 2 hours of ischemia and then untwisted for 2 hours of reperfusion. The control ponies underwent 4 hours of anesthesia. Full-thickness colonic biopsy specimens were obtained from the antimessenteric border of the pelvic flexure and prepared for light and transmission electron microscopy at 0, 1, 2, 3, and 4 hours after twisting. The mean histopathology score at 0 hours for control ponies was 1.2 and remained unchanged; the score for the experimental ponies was 1.8 and progressed to 4.2 by 4 hours. At 2 hours of ischemia, there was edema of the submucosa and lamina propria attributable to accumulation of platelets, fibrin, and neutrophils in capillaries of the lamina propria. The morphologic changes in experimental ponies progressed to complete destruction of the mucosa with congestion, hemorrhage, and microvascular thrombosis. Mean histopathology scores of control and experimental ponies were significantly different at 1 hour of ischemia, and lesions progressively worsened through the reperfusion period. The morphologic changes observed in this study accurately reflect those changes reported in horses with naturally occurring colonic volvulus and can serve as a reference for subsequent studies on attenuating bowel injury.
Immunohistochemical Demonstration of Parathyroid Hormone-related Protein in Thyroid Gland of Sheep
Authors: Okada H. , Capen C. C. , Rosol T. J.
Published: January 2010

Thyroid and parathyroid glands from normal adult sheep and sheep with vitamin D3-induced hypercalcemia were evaluated immunohistochemically for the presence of parathyroid hormone-related protein (PTHrP) and calcitonin (CT). Thyroid follicular cells, C-cells, and parathyroid chief cells stained moderately to strongly positive for PTHrP in both groups of sheep. C-cells were identified as cells positive for cytoplasmic staining of CT. Staining of C-cells for calcitonin in hypercalcemic sheep was diminished in intensity. The data demonstrated that PTHrP is present in the sheep thyroid and parathyroid glands. PTHrP may function as a local paracrine factor in these organs, but its staining intensity was not affected by the induction of hypercalcemia.
Basal Cell Tumor in a Japanese Macaque (Macaca fuscata)
Authors: Yanai T. , Wakabayashi S. , Masegi T. , Iwasaki T. , Yamazoe K. , Ishikawa K. , Ueda K.
Published: January 2010

A rare case of basal cell tumor of the skin in a wild-caught female Japanese macaque ( Macaca fuscata) was studied. Tumor growth, which was composed of cells resembling basal cells, was observed in the dermis and subcutis and showed a mixture of ribbon, solid, glandular, and cystic patterns. The tumor was separated from surrounding normal tissue by a compressed zone of fibrous connective tissue. The nuclei were round and hyperchromatic and possessed a single centrally located nucleolus. Mitotic figures were common. Immunohistochemically, a positive reaction for epithelial membrane antigen, keratin, and cytokeratin AE3 was evident. Electron microscopic examination revealed a small number of tonofilaments in the cytoplasm.
Early Ultrastructural Glomerular Alterations in Neonatal Nephrotic Mice (ICGN Strain)
Authors: Ogura A. , Fujimura H. , Asano T. , Koura M. , Naito I. , Kobayashi Y.
Published: January 2010

ICGN is a strain of mice with hereditary nephrotic syndrome of an unknown cause. In this study, early glomerular alterations in newborn ICGN mice were observed with electron microscopy to gain a better insight into the onset of the disease. Development of the glomeruli was normal until fusion of epithelial and endothelial basement membranes in the developing capillary stage. From the maturing glomerulus stage onward, the fused glomerular basement membrane (GBM) increased in thickness by excessive accumulation of the basement membrane material secreted from the epithelial cells. This accumulation was followed by overall loss of epithelial foot processes in the glomeruli. These findings indicate that the disease in ICGN mice is caused by some defect(s) in the GBM maturation process, which may be crucial for the generation of the glomerular permselectivity.
The Early Dynamic Response of the Calf Ileal Epithelium toSalmonella typhimurium
Authors: Frost A. J. , Bland A. P. , Wallis T. S.
Published: January 2010

Ileal loops including Peyer's patch were prepared in five 28-day-old calves and infused with Salmonella typhimurium strain ST4/74. Loops were fixed 5 minutes to 2 hours after inoculation, and the mucosa was examined by light and electron microscopy. Within 5 minutes, the bacteria were interacting with the follicle-associated epithelium (FAE); the surface of M cells changed to lamellipodia, engulfing many bacteria. This process proceeded rapidly to 30 minutes, involving most M cells above crypt level. Most cells were exfoliated, and many were packed with bacteria, and the domed villi became stunted. There was a rapid migration of neutrophils through the FAE into the lumen by 15 minutes. By 60 minutes, there was no further interaction between the bacteria and the FAE; at this time bacteria were present in macrophages in the lamina propria. Restitution of the FAE was complete by 2 hours in spite of the many bacteria in the cell debris overlying the epithelium. Interaction of bacteria with the absorptive villi was delayed compared with interaction with the FAE. After 15 minutes, bacteria were seen adhering to some enterocytes of the upper third of the villi; many bacteria were adhering to the surface of the enterocytes at 20 and 30 minutes, but few were seen thereafter. Adherence was patchy and largely confined to cells whose surfaces were depressed relative to others. The microvillous surface of these enterocytes was extensively remodelled. Tissue response, with uptake of bacteria into vacuoles, exfoliation of enterocytes containing bacteria, and subsequent stunting of the villi, began at 30 minutes and was severe and progressive to 2 hours. Following the initial attachment and uptake of the bacteria, loss of enterocytes progressed from these initial sites; bacteria were associated with the lateral cell membrane of cells adjacent to cells being extruded and not with the microvilli of cells at new sites. In a calf 4 hours after dosing orally with the same strain, M cells were engulfing bacteria and their cell surface was changed as seen in the inoculated loops; absorptive enterocytes were also taking up bacteria as seen in the ileal loops, indicating the process seen in the loops and after oral dosage was similar. For this strain of S. typhimurium, there was an initial concentration of bacilli around the domed villus epithelium. This distribution was not random but may have resulted from a specific attraction to the FAE.
Immunohistochemistry of Pancreatic Islet Cell Tumors in the Ferret (Mustela putorius furo)
Authors: Andrews G. A. , Myers N. C. , Chard-Bergstrom C.
Published: January 2010

Twenty-two pancreatic islet cell tumors and normal pancreatic islets from ferrets were evaluated by immunohistochemistry for expression of the peptide hormones insulin, somatostatin, glucagon, and pancreatic polypeptide (PP) and the neuroendocrine markers chromogranin A (CgA) and neuron-specific enolase (NSE). In normal pancreatic islets, the majority of cells stained strongly with CgA and NSE. A cells, B cells, D cells, and PP cells stained strongly with glucagon, insulin, somatostatin, and PP, respectively. All 22 tumors stained with CgA and NSE. The proportion of cells within tumors staining for CgA was variable, but more than half of the cells stained positively in 18 of the tumors. The intensity of staining for CgA was strong (reactivity equivalent to or greater than normal islet cells in adjacent tissue) in 11 moderate in six, and weak in five of the tumors. All tumors stained for NSE, with ≥50% of the cells staining in 21 of the tumors, and the intensity of staining was strong in 18 of the tumors. Twenty of 22 tumors stained positively for insulin, with ≥50% of the cells staining in 19 of them. The intensity of staining for insulin was strong in 12, moderate in seven, and weak in one of the tumors. Approximately ≤1% of the cells in 15 of 22 tumors stained for somatostatin, five tumors stained for pancreatic polypeptide, and three tumors stained for glucagon. These data indicate that the majority of islet cell tumors of ferrets express immunohistochemically detectable insulin. CgA and NSE are both useful general markers for such tumors, including those that are insulin negative. Commercially available antisera to CgA, NSE, insulin, glucagon, somatostatin, and PP work well in formalin-fixed, paraffin-embedded tissue for immunophenotyping islet cell tumors in the ferret.
Fatal Disseminated Cercopithecine Herpesvirus 1 (Herpes B) Infection in Cynomolgus Monkeys {Macaca fascicularis)
Authors: Carlson C. S. , O'Sullivan M. G. , Jayo M. J. , Anderson D. K. , Harber E. S. , Jerome W. G. , Bullock B. C. , Heberling R. L.
Published: January 2010

Two adult female cynomolgus monkeys ( Macaca fascicularis) that had been housed together for 4 months died within 2 weeks of each other after brief illnesses. Monkey No. 1 presented with collapse, watery stool, and hypothermia and died overnight. Monkey No. 2 presented with dyspnea, nasal discharge, leukopenia, and hypoproteinemia and was euthanized after 2 days. Both animals had peritoneal effusions, massive necrosis of pharyngeal, esophageal, and gastric mucosa, and multifocal hepatic and pancreatic necrosis. Monkey No. 2 also had lingual ulcers and locally extensive necrosis of spleen, adrenal glands, and lymph nodes. Large numbers of eosinophilic intranuclear inclusion bodies were present in epithelial and syncytial cells adjoining the necrotic foci in Monkey No. 2 but were absent in Monkey No. 1. Monkey No. 1 seroconverted to cercopithecine herpesvirus 1 (CHV-1, commonly known as herpes B) in the month before death. CHV-1 was isolated from a sample of stomach from Monkey No. 2, and electron microscopy of liver from this animal demonstrated herpesvirus particles within hepatocytes. Both animals were seropositive for simian type D retrovirus, and the virus was cultured from the liver of Monkey No. 2. A diagnosis of disseminated CHV-1 infection was made, possibly occurring secondary to immunosuppression due to infection with simian type D retrovirus. Although a high percentage of cynomolgus monkeys are inapparently infected with CHV-1, disseminated disease is rare. Because infection with CHV-1 in humans is associated with a high fatality rate, familiarity with the lesions of disseminated infection with this virus is important.
Effects ofPasteurella multocidaToxin on Porcine Bone Marrow Cell Differentiation into Osteoclasts and Osteoblasts
Authors: Gwaltney S. M. , Galvin R. J. S. , Register K. B. , Rimler R. B. , Ackermann M. R.
Published: January 2010

The effect of Pasteurella multocida toxin (PMT) on porcine osteoclast and osteoblast differentiation was studied using in vitro cell culture systems. When grown in the presence of Vitamin D3, isolated porcine bone marrow cells formed multinucleated cells with features characteristic of osteoclasts. Exposure of bone marrow cells to Vitamin D3and PMT during growth resulted in formation of increased numbers and earlier appearance of osteoclasts compared to controls. Ultrafiltered medium from PMT-treated cells likewise increased osteoclast numbers, suggesting that a soluble mediator may be involved in the action of PMT. When cell cultures were treated with fluorescein-labeled PMT, fluorescence was found within the cytoplasm of small, round cells that did not resemble either osteoclasts or osteoclastic precursor cells.

Cultures of porcine bone marrow cells exposed to dexamethasone, ascorbic acid, and β-glycerophosphate developed into osteoblastic cells that formed multilayered, mineralized nodules. Exposure of osteoblastic cultures to low concentrations of PMT resulted in retarded cell growth, formation of decreased numbers of nodules, and minimal to no mineralization in the nodules; higher concentrations of PMT resulted in increased cellular debris and poor growth of cells, with no nodule formation. These findings suggest that PMT may induce turbinate atrophy in pigs by increasing osteoclast numbers and inhibiting osteoblastic bone formation. The effect of PMT on osteoclastic differentiation and growth may not be due to a direct effect on preosteoclastic cells, but rather due to alterations in the soluble mediator secretion by marrow stromal cells.
Pulmonary Lesions in Experimental Ophidian Paramyxovirus Pneumonia of Aruba Island Rattlesnakes,Crotalus unicolor
Authors: Jacobson E. R. , Adams H. P. , Geisbert T. W. , Tucker S. J. , Hall B. J. , Homer B. L.
Published: January 2010

Histologic and ultrastructural changes were observed in the respiratory portions of lung in five 29-40-month-old Aruba Island rattlesnakes, Crotalus unicolor, that were inoculated with an Aruba Island Rattlesnake virus (ATV) strain of ophidian paramyxovirus (OPMV) isolated from an Aruba Island rattlesnake. Lungs from one non-infected and three mock-infected Aruba Island rattlesnakes were examined also. From 4 to 22 days following intratracheal inoculation, progressive microscopic changes were seen in the lung. Initially, increased numbers of heterophils were observed in the interstitium followed by proliferation and vacuolation of epithelial cells lining faveoli. The changes appeared to progress from cranial to caudal portions of the respiratory lung following inoculation. Beginning at 4 days postinoculation, viral antigen was demonstrated in epithelial cells lining faveoli with an immunofluorescent technique using a rabbit anti-AIV polyclonal antibody. Electron microscopy revealed loss of type I cells, hyperplasia of type II cells, and interstitial infiltrates of heterophils and mononuclear cells. Viral nucleocapsid material was seen within the cytoplasm and mature virus was seen budding from cytoplasmic membranes of infected type I and type II cells from 8 to 19 days after infection. A virus consistent with AIV was isolated from lung tissues of infected rattlesnakes, thus fulfilling Koch's postulates.