Journal of Comparative Pathology

Bibliography review

Canine mammary tumors.....................................................................................................................1 Diverse tumors......................................................................................................................................2 Viruses..................................................................................................................................................4 Articles ................................................................................................................................................5 Circovirus...........................................................................................................................................11 Maedi-Visna.......................................................................................................................................12 Scrapie................................................................................................................................................13 PRRS..................................................................................................................................................16 Distemper...........................................................................................................................................17 Foot-and-Mouth..................................................................................................................................17

Canine mammary tumors
Author: Restucci, B.; Borzacchiello, G.; Maiolino, P.; Martano, M.; Paciello, O.; Papparella, S. Year: 2004 Title: Expression of vascular endothelial growth factor receptor Flk-1 in canine mammary tumours Journal: J Comp Pathol Volume: 130 Issue: 2-3 Pages: 99-104 Date: Feb-Apr Abstract: Vascular endothelial growth factor (VEGF) induces endothelial cell proliferation, and the beginning of angiogenesis, by interacting with specific endothelial receptors termed VEGFR-1 (Flt1) and VEGFR-2 (Flk-1). In this study, Flk-1 expression was evaluated immunohistochemically in 10 benign and 40 malignant canine mammary tumours. There was immunolabelling of endothelial cells located within the neoplastic proliferation and at the infiltrating periphery, and also of neoplastic cells. The number of positive endothelial and neoplastic cells, was higher in malignant than in benign tumours. Moreover, in the malignant tumours, expression of Flk-1 increased from well to less differentiated phenotypes (grade 1-3). The presence of VEGF receptor on neoplastic cells suggests that VEGF has an autocrine function in which neoplastic cells act as both VEGF producers and target cells. Thus, in malignant tumours, VEGF may contribute to neoplastic growth by inducing angiogenesis and by stimulating the proliferation of neoplastic cells.

Author: Gama, A.; Paredes, J.; Albergaria, A.; Gartner, F.; Schmitt, F. Year: 2004 Title: P-cadherin expression in canine mammary tissues Journal: J Comp Pathol Volume: 130 Issue: 1 Pages: 13-20 Date: Jan Accession Number: 14693120 Abstract: P-cadherin is a classical cadherin expressed by myoepithelial cells in mammary tissue. Its expression in human breast cancer has been associated with aggressive tumour behaviour. To analyse the possible role of P-cadherin in canine mammary carcinogenesis, its expression was

examined immunohistochemically in 82 samples of normal (n=2), hyperplastic (n=11) and neoplastic (n=69) canine mammary tissues. In normal and hyperplastic canine mammary glands, P-cadherin was restricted to myoepithelial cells, usually at sites of cell-to-cell contact. In tumour tissues, however, P-cadherin expression was observed in both epithelial and myoepithelial cells, with a cytoplasmic pattern of cellular distribution. Aberrant epithelial P-cadherin immunolabelling was found in 19/44 (43%) benign tumours and in 16/25 (64%) malignant tumours (P<0.001). In malignant tumours, a significant correlation between P-cadherin expression intensity and histological type was observed (P<0.05).

Author: Matos, A. J.; Lopes, C.; Carvalheira, J.; Santos, M.; Rutteman, G. R.; Gartner, F. Year: 2006 Title: E-cadherin expression in canine malignant mammary tumours: relationship to other clinico-pathological variables Journal: J Comp Pathol Volume: 134 Issue: 2-3 Pages: 182-9 Date: Feb-Apr Abstract: The relationship between E-cadherin epithelial expression, as detected by immunohistochemical methods, and other clinico-pathological characteristics of canine malignant mammary tumours was studied in 77 tumours surgically removed from 45 female dogs. The immunohistochemical assessment was based on the estimated percentage of epithelial cells with membranous labelling. Reduction of E-cadherin expression was significantly related to size and ulceration of tumours but not to fixation to skin or underlying tissue; it was also related to lymph node metastasis, necrosis and infiltrative growth. Histological type (but not histological grade) was related to E-cadherin expression, with solid tumours more frequently lacking expression and tubulopapillary tumours showing increased expression as compared with the other types. The significant relationship between E-cadherin and other known factors of poor prognosis suggests that the loss of E-cadherin expression may have prognostic value in canine malignant mammary tumours.

Diverse tumors
Author: Machida, N.; Tanaka, R.; Takemura, N.; Fujii, Y.; Ueno, A.; Mitsumori, K. Year: 2004 Title: Development of pericardial mesothelioma in golden retrievers with a long-term history of idiopathic haemorrhagic pericardial effusion Journal: J Comp Pathol Volume: 131 Issue: 2-3 Pages: 166-75 Date: Aug-Oct Abstract: This report describes the development of pericardial mesothelioma in five golden retrievers with a long-term history of idiopathic haemorrhagic pericardial effusion (IHPE). These five dogs were treated with repeated pericardiocentesis for recurrent episodes of pericardial fluid accumulation; other than IHPE, all potential causes of this fluid accumulation were ruled out by the results of diagnostic imaging and cytology and bacterial or fungal culture of fluid obtained during pericardiocentesis. In three dogs that eventually underwent pericardiectomy, neoplastic lesions were not detected in any organs or tissues within the thoracic cavity during the surgical procedure, and the surgical biopsies were consistent with IHPE. In one of the three dogs, however, cytology of recurrent thoracic effusion revealed clusters of neoplastic mesothelial cells from 1 month after surgical intervention until death. The clinical course of the disease ranged from 30 to 54 months

between the first visit and death, and on post-mortem examination pericardial mesothelioma was diagnosed in all five dogs. The clinical observations, together with the breed and age of the affected animals, suggested that the five dogs initially suffered from IHPE, which was then followed by the development of pericardial mesothelioma. It is possible that IHPE is associated with the development of pericardial mesothelioma in golden retrievers through a chronic inflammatory process.

Author: Ueki, H.; Kowatari, Y.; Oyamada, T.; Oikawa, M.; Yoshikawa, H. Year: 2004 Title: Non-functional C-cell adenoma in aged horses Journal: J Comp Pathol Volume: 131 Issue: 2-3 Pages: 157-65 Date: Aug-Oct Abstract: Thyroid tumours occur in older horses, and most such tumours have been considered to be of follicular epithelial origin. However, their immunohistochemical characterization has not been investigated. The purpose of this study was to confirm a suspicion that most of these tumours are in fact parafollicular cell (C cell)-derived adenomas, and to evaluate their pathogenesis and functional state. Thyroid glands from 38 horses aged 10-29 years were evaluated, all tissue samples being examined histologically, immunohistochemically and ultrastructurally. Nodular tumour masses were found in the thyroids of 12 of 38 horses older than 10 years (31.6%), and in nine of 12 horses older than 20 years (75.0%), regardless of sex or breed. Nodular lesions were composed of solid proliferations of polygonal cells with eosinophilic granular cytoplasm. Immunohistochemically, tumour cells were positive for calcitonin and neuron specific enolase, but negative for thyroglobulin. Ultrastructurally, few if any secretion granules were found in tumour cells. On the basis of these results it was concluded that the nodular lesions were C-cell adenomas, not follicular adenomas. It was suspected that the C-cell adenomas were nonfunctional and unlikely to lead to calcitonin hypersecretion-related diseases.

Author: Day, M. J.; Henderson, S. M.; Belshaw, Z.; Bacon, N. J. Year: 2004 Title: An immunohistochemical investigation of 18 cases of feline nasal lymphoma Journal: J Comp Pathol Volume: 130 Issue: 2-3 Pages: 152-61 Date: Feb-Apr Accession Number: 15003473 Abstract: This report details clinical, histopathological and immunohistochemical findings in 18 cats with chronic nasal disease diagnosed as nasal lymphoma. Eight of the cats were female and 10 were male, with a median age of 10.5 years (range 7-14 years). Three of the cats were Siamese, one was Burmese, and the rest were non-pedigree. The duration of clinical signs before referral ranged from 30 to 540 days (median 88.5 days). The most common clinical signs were nasal discharge, stertor and sneezing. Nasal radiographs were abnormal in 14/16 cases examined. Abnormal masses were detected endoscopically in 13/18 cases. Nine cats received multi-agent chemotherapy or radiation therapy, or both, with survival times ranging from 14 to >541 days. Biopsy material from these 18 cats was examined by light microscopy, and serial sections were subjected to immunohistochemical labelling for the T lymphocyte marker CD3 and the B lymphocyte marker CD79a. In 13 tissues, expression of class II molecules of the major histocompatibility complex and the myelomonocytic antigen MAC387 was also determined. Twelve of the tumours were classified as diffuse large B-cell lymphomas, four as lymphoblastic B-cell lymphomas, and one as a follicular B-cell lymphoma. The tumour cells within these lesions all

expressed CD79a, and (where tested) most also expressed MHC class II. One tumour was an anaplastic large cell neoplasm, in which the neoplastic cells expressed MHC class II alone in the absence of either lymphoid marker. There was a variable infiltration of reactive small T lymphocytes into these tumours, and zones of necrosis within the tumour tissue were sometimes heavily infiltrated by MAC387+ phagocytic cells.

Author: Waly, N. E.; Gruffydd-Jones, T. J.; Stokes, C. R.; Day, M. J. Year: 2005 Title: Immunohistochemical diagnosis of alimentary lymphomas and severe intestinal inflammation in cats Journal: J Comp Pathol Volume: 133 Issue: 4 Pages: 253-60 Date: Nov Accession Number: 16213517 Abstract: Intestinal tissue samples were examined from 32 cats in which a histopathological diagnosis of alimentary lymphoma or multicentric lymphoma affecting the gastrointestinal tract had been made. These samples were re-evaluated histopathologically and serial sections were examined immunohistochemically with antisera specific for the lymphoid markers CD3, CD79a and BLA-36 and for class II molecules of the major histocompatability complex. The cats ranged in age from 4-16 years (median 10.5 years). The main presenting clinical signs were vomiting, diarrhoea and weight loss. The majority of alimentary lymphomas were of the B-cell type (n=15), whereas cases of T-cell lymphoma were fewer in number (n=8). Four cats had lymphoma of a mixed T-and B-cell phenotype. In five of the cats, immunohistochemistry suggested an inflammatory process, in contradiction to the original histopathological diagnosis of lymphoma. Immunolabelling would appear to be a useful adjunct to histopathology in classifying cases of feline alimentary lymphoma, and may help in distinguishing lymphoma from severe intestinal inflammation.

Viruses
Author: Ortin, A.; Perez de Villarreal, M.; Minguijon, E.; Cousens, C.; Sharp, J. M.; De las Heras, M. Year: 2004 Title: Coexistence of enzootic nasal adenocarcinoma and jaagsiekte retrovirus infection in sheep Journal: J Comp Pathol Volume: 131 Issue: 4 Pages: 253-8 Date: Nov Abstract: Ten sheep naturally affected with enzootic nasal adenocarcinoma (ENA), a disease associated with ovine enzootic nasal tumour virus (ENTV-1), were found also to be infected with jaagsiekte sheep retrovirus (JSRV), the causal agent of ovine pulmonary adenocarcinoma (OPA). Only one of the sheep showed OPA lung lesions. The animals belonged to 10 flocks located in a geographical area in which OPA is frequently seen. ENTV-1 was found in all the ENA tumours but only occasionally in extra-tumoral sites, confirming the results of a previous study. In contrast, JSRV had a disseminated tissue distribution, similar to that previously reported for animals infected with JSRV. However, the occurrence of JSRV in lymphoid tissues was clearly greater than in sheep infected with JSRV but with no lesions of ENA. The data suggested a synergistic relationship between ENTV-1 and JSRV, resulting in increased proliferation of JSRV.

Author: Tjornehoj, K.; Fooks, A. R.; Agerholm, J. S.; Ronsholt, L. Year: 2006 Title: Natural and experimental infection of sheep with European bat lyssavirus type-1 of Danish bat origin Journal: J Comp Pathol Volume: 134 Issue: 2-3 Pages: 190-201 Date: Feb-Apr Accession Number: 16545840 Abstract: In 1998 and 2002, European bat lyssavirus type-1 (EBLV-1) was demonstrated in brain tissue of five Danish sheep suffering from neurological disorders. Four of the five sheep also had encephalic listeriosis. The animals originated from four flocks on pastures within a limited area of western Jutland. In a serological investigation in two of the herds, from which three of the diseased animals originated, EBLV-1 neutralizing antibodies were detected in only one of 69 sheep. In follow-up surveys, 2110 sheep sera collected at Danish slaughterhouses during 2000 were all negative for EBLV-1-antibodies, and EBLV-1 was not demonstrated in 87 ruminants displaying neurological symptoms. To investigate the pathogenic effects of EBLV-1, four sheep were inoculated intralabially with either brain material from one of the naturally infected sheep or virus isolated from the same sheep. These animals developed EBLV-1 neutralizing antibodies at 5-9 weeks post-inoculation but did not exhibit neurological signs during a 33-week observation period. It was speculated that the immune response prevented viral dissemination to the brain, resulting in an abortive peripheral infection. It was concluded that EBLV-1 can infect sheep under natural conditions as an incidental event.

Articles
Author: Dubey, J. P.; Buxton, D.; Wouda, W. Year: 2006 Title: Pathogenesis of bovine neosporosis Journal: J Comp Pathol Volume: 134 Issue: 4 Pages: 267-89 Date: May Abstract: The protozoan parasite Neospora caninum is a major pathogen of cattle and dogs, being a significant cause of abortion in cattle in many countries. It is one of the most efficiently transmitted parasites, with up to 90% of cattle infected in some herds. The pathogenesis of abortion due to Neospora is complex and only partially understood. Losses occur after a primary infection during pregnancy but more commonly as the result of recrudescence of a persistent infection during pregnancy. Parasitaemia is followed by invasion of the placenta and fetus. It is suggested that abortion occurs when primary parasite-induced placental damage jeopardises fetal survival directly or causes release of maternal prostaglandins that in turn cause luteolysis and abortion. Fetal damage may also occur due to primary tissue damage caused by the multiplication of N. caninum in the fetus or due to insufficient oxygen/nutrition, secondary to placental damage. In

addition, maternal immune expulsion of the fetus may occur associated with maternal placental inflammation and the release of maternal pro-inflammatory cytokines in the placenta. Thus N. caninum is a primary pathogen capable of causing abortion either through maternal placental inflammation, maternal and fetal placental necrosis, fetal damage, or a combination of all three. The question of how N. caninum kills the fetus exposes the complex and finely balanced biological processes that have evolved to permit bovine and other mammalian pregnancies to occur. Defining these immunological mechanisms will shed light on potential methods of control of bovine neosporosis and enrich our understanding of the continuity of mammalian and protozoal survival. Author: Choi, C.; Kwon, D.; Jung, K.; Ha, Y.; Lee, Y. H.; Kim, O.; Park, H. K.; Kim, S. H.; Hwang, K. K.; Chae, C. Year: 2006 Title: Expression of inflammatory cytokines in pigs experimentally infected with Mycoplasma hyopneumoniae Journal: J Comp Pathol Volume: 134 Issue: 1 Pages: 40-6 Date: Jan Abstract: The expression of interleukin-1 (IL-1), tumour necrosis factor-alpha (TNF-alpha) and IL-6 were studied over a period of 35 days in the lungs of pigs experimentally infected with Mycoplasma hyopneumoniae, by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR), morphometric analysis and in-situ hybridization. Fifteen colostrum-deprived pigs aged 14 days were inoculated intranasally with M. hyopneumoniae. IL-1, TNF-alpha and IL-6 were detected by RT-PCR in the lungs of the infected pigs from 7 days post-inoculation (dpi) onwards, but not in the uninfected control pigs. Concurrent expression of all three cytokines was always observed, in association with lung lesions. Inflammatory cytokine-positive cells were detected in the lungs at 7 dpi, their number increasing at 21dpi, and decreasing thereafter. The results suggest that such cytokines play a role in mediating and regulating inflammation in M. hyopneumoniae infection. Author: Cecchi, R.; Wills, S. J.; Dean, R.; Pearson, G. R. Year: 2006 Title: Demonstration of Ollulanus tricuspis in the stomach of domestic cats by biopsy Journal: J Comp Pathol Volume: 134 Issue: 4 Pages: 374-7 Date: May Abstract: Ollulanus tricuspis is a small nematode of the family Ollulanidae, found in the stomach of domestic cats and other felids. Of 131 gastric biopsy samples collected at endoscopic examination, four were shown to contain the parasite. Vomiting was the main presenting sign in three cats and weight loss in the fourth. The stomachs were grossly normal on endoscopic examination. Chronic gastritis was observed histologically in two cases, while the remaining cases were microscopically normal. The significance of the parasite remained undetermined. To our knowledge, this is the first report of O. tricuspis infection in domestic cats in which the diagnosis was made by examining routine endoscopic biopsy samples. Author: Carvalho, T.; Pinto, C.; Peleteiro, M. C. Year: 2006 Title: Urinary bladder lesions in bovine enzootic haematuria Journal: J Comp Pathol Volume: 134 Issue: 4 Pages: 336-46 Date: May Abstract: In cattle, bracken fern chronic toxicity is characterized by the presence of multiple tumours in the bladder (bovine enzootic haematuria). From October 1999 to March 2003, 433

urinary bladders with macroscopical lesions were collected in the slaughterhouse of Sao Miguel Island (Azores, Portugal), an endemic area where Pteridium aquilinum infestation in pastures is high. Bladder lesions were divided into three main categories (inflammatory lesions, non-neoplastic epithelial abnormalities and tumours) and described in detail. In some cases, neoplastic growth was confined to a single site, but in most cases multiple tumours developed within the same bladder. Epithelial tumours alone were present in 51.2% of the affected bladders, mesenchymal tumours alone in 17.4%, and both epithelial and mesenchymal tumours in the remaining 31.4%. The large number of tumours examined (870) revealed new categories not yet included in other veterinary classification systems, namely, inverted papilloma, papillary neoplasm of apparent low malignant potential, and haemangioendothelioma.

Author: Wangoo, A.; Johnson, L.; Gough, J.; Ackbar, R.; Inglut, S.; Hicks, D.; Spencer, Y.; Hewinson, G.; Vordermeier, M. Year: 2005 Title: Advanced granulomatous lesions in Mycobacterium bovis-infected cattle are associated with increased expression of type I procollagen, gammadelta (WC1+) T cells and CD 68+ cells Journal: J Comp Pathol Volume: 133 Issue: 4 Pages: 223-34 Date: Nov Abstract: The pathognomonic characteristic of tuberculosis (TB) is the formation of a tuberculous granuloma. The objective of this study was to classify lymph node granulomas from experimentally infected calves into different histopathological stages and characterize them further by studying cell types and markers of fibrosis associated with each of the stages. Four stages of granuloma were identified and mRNA and protein expression for cell markers, cytokines and pro-fibrotic markers were studied by immunohistochemistry (IHC) and in-situ hybridization (ISH). In advanced stage granulomas, there was an increase in the expression of TGF-beta, and of type I procollagen as demonstrated by IHC and ISH. As the granulomas advanced, there were fewer CD3+T cells and they tended to be more prominent towards the periphery of the lesions, with a steady increase in the number of CD68+ cells and gammadelta (WC1+) T cells. Granuloma classification and application of cell cytokine markers will assist in improving understanding of the pathogenesis of bovine TB and may help to identify the immunopathology of active disease versus contained or inactive disease. Such disease correlates may help to inform the development of improved diagnostic methods and support vaccine development programmes. Year: 2005 Title: Attaching-effacing bacteria in animals Journal: J Comp Pathol Volume: 132 Issue: 1 Pages: 1-26 Date: Jan Abstract: Enteric bacteria with a demonstrable or potential ability to form attaching-effacing lesions, so-called attaching-effacing (AE) bacteria, have been found in the intestinal tracts of a wide variety of warm-blooded animal species, including man. In some host species, for example cattle, pigs, rabbits and human beings, attaching-effacing Escherichia coli (AEEC) have an established role as enteropathogens. In other host species, AE bacteria are of less certain significance. With continuing advances in the detection and typing of AE strains, the importance of these bacteria for many hosts is likely to become clearer. The pathogenic effects of AE bacteria result from adhesion to the intestinal mucosa by a variety of mechanisms, culminating in the formation of the characteristic intimate adhesion of the AE lesion. The ability to induce AE lesions is mediated by

the co-ordinated expression of some 40 bacterial genes organized within a so-called pathogenicity island, known as the "Locus for Enterocyte Effacement". It is also believed that the production of bacterial toxins, principally Vero toxins, is a significant virulence factor for some AEEC strains. Recent areas of research into AE bacteria include: the use of Citrobacter rodentium to model human AEEC disease; quorum-sensing mechanisms used by AEEC to modulate virulence gene expression; and the potential role of adhesion in the persistent colonization of the intestine by AE bacteria. This review of AE bacteria covers their molecular biology, their occurrence in various animal species, and the diagnosis, pathology and clinical aspects of animal diseases with which they are associated. Reference is made to human pathogens where appropriate. The focus is mainly on natural colonization and disease, but complementary experimental data are also included.

Author: Summers, B. A.; Straubinger, A. F.; Jacobson, R. H.; Chang, Y. F.; Appel, M. J.; Straubinger, R. K. Year: 2005 Title: Histopathological studies of experimental lyme disease in the dog Journal: J Comp Pathol Volume: 133 Issue: 1 Pages: 1-13 Date: Jul Accession Number: 15904927 Abstract: Experimental borrelia infection was induced in 62 specific--pathogen-free beagle dogs by exposure to Ixodes scapularis ticks harbouring the spirochaete Borrelia burgdorferi. Clinical signs of Lyme disease occurred in 39/62 dogs, the remaining 23 being subclinically infected. Clinical signs consisted of one to six episodes of transitory lameness with joint swelling and pain, most commonly affecting the elbow or shoulder joints. The polymerase chain reaction and culture demonstrated that the dogs remained infected for up to 581 days. At necropsy, gross findings consisted of lymphadenopathy in the area of tick attachment. Microscopical changes consisted of effusive fibrinosuppurative inflammation or nonsuppurative inflammation, or both, affecting synovial membranes, joint capsules and associated tendon sheaths. Plasma cells dominated areas of chronic inflammation, with CD3(+) T cells being present in lesser numbers. Microscopical signs of arthritis were polyarticular and more widespread than indicated by clinical signs, and most of the subclinically affected animals also had synovitis. In areas of tick attachment to the skin, hyperkeratosis and a mixture of suppurative and nonsuppurative dermatitis were encountered. Lymphadenopathy in superficial lymph nodes resulted from follicular and parafollicular hyperplasia. In 14/62 dogs, lymphoplasmacytic periarteritis and perineuritis were noted, resembling lesions found in human Lyme disease and syphilis, in which an underlying microangiopathy has been proposed.

Author: Rallis, T.; Day, M. J.; Saridomichelakis, M. N.; Adamama-Moraitou, K. K.; Papazoglou, L.; Fytianou, A.; Koutinas, A. F. Year: 2005

Title: Chronic hepatitis associated with canine leishmaniosis (Leishmania infantum): a clinicopathological study of 26 cases Journal: J Comp Pathol Volume: 132 Issue: 2-3 Pages: 145-52 Date: Feb-Apr Accession Number: 15737341 Abstract: Hepatic tissue samples were obtained from 26 dogs humanely destroyed because of naturally occurring leishmaniosis (Leishmania infantum). None of the animals had palpable hepatomegaly or any other physical finding or historical evidence indicative of liver failure. However, serum biochemistry revealed hypoalbuminaemia (6/26), increased alkaline phosphatase (ALP) activity (15/26), and increased concentrations of total bilirubin (2/26) and post-prandial bile acids (4/26). Three main histological patterns were identified. In pattern 1 (3/26), the liver microarchitecture remained unchanged apart from the presence of individual or clustered macrophages in the sinusoids. In pattern 2 (20/26), there was multifocal, mild to moderate, granulomatous to pyogranulomatous infiltration of the hepatic parenchyma, particularly in the portal areas. Pattern 3 (3/26), which was the most severe form, was characterized by marked portal lymphoplasmacytic infiltration with occasional broaching of the limiting plate and extension into the adjacent parenchyma. In this pattern there was also mild portal fibrosis, together with lymphoplasmacytic aggregates within the parenchyma and small clusters of lymphocytes and plasma cells within the sinusoids. All three patterns were associated with hepatocyte vacuolation (15/26 dogs), and haemosiderin accumulation within the hepatocyte cytoplasm. Congestion was present in the liver of five dogs. No correlation was found between histopathological pattern and breed, sex, age, clinical manifestations, serum biochemical profile or parasite load in the hepatic tissue; patterns 1-3 may, however, represent sequential stages of hepatic leishmania infection during the chronic course of the disease. Author: Peeters, D.; Day, M. J.; Clercx, C. Year: 2005 Title: Distribution of leucocyte subsets in bronchial mucosa from dogs with eosinophilic bronchopneumopathy Journal: J Comp Pathol Volume: 133 Issue: 2-3 Pages: 128-35 Date: Aug-Oct Abstract: Immunohistochemistry was used to characterize the distribution of leucocyte subsets in the bronchial mucosa of 11 dogs with idiopathic eosinophilic bronchopneumopathy (EBP). Formalin-fixed tissues from all dogs were included in the study, but frozen tissue from only one dog was available. MHC class II(+) cells were found in moderate numbers in the lamina propria (LP). These cells were morphologically either dendritic-like cells or macrophages, but many macrophages did not express MHC class II. Such molecules were expressed by occasional fibroblasts. L1(+) cells, which formed a relatively small component of the LP inflammatory infiltrate, were morphologically either macrophages or polymorphonuclear cells (probably eosinophils). IgA(+) plasma cells were found in varying numbers in the LP, mostly in association with glandular tissue. IgG(+) plasma cells were less common, and IgM(+) plasma cells were present in low numbers. Many CD3(+) cells were present in the LP. In the single case from which frozen tissue was available, most of the lymphocytes were labelled with CD4 marker, while smaller numbers were CD8(+) T cells. Most of the lymphocytes in this case were positively labelled with T-cell receptor (TCR)-alphabeta marker. TCR-gammadelta(+) cells, although less common, were present in significant numbers throughout the LP. CDlc(+) dendritic cells were numerous in the epithelium and in the LP, immediately beneath the basement membrane. These findings, which were similar to those described in human asthma, are suggestive of a Th2 dominant immune response in canine EBP. As in human asthma, this provides a possible basis for new forms of treatment in canine EBP.

Author: Gonzalez, J.; Geijo, M. V.; Garcia-Pariente, C.; Verna, A.; Corpa, J. M.; Reyes, L. E.; Ferreras, M. C.; Juste, R. A.; Garcia Marin, J. F.; Perez, V. Year: 2005 Title: Histopathological classification of lesions associated with natural paratuberculosis infection in cattle Journal: J Comp Pathol Volume: 133 Issue: 2-3 Pages: 184-96 Date: Aug-Oct Abstract: Paratuberculosis-associated lesions in 116 naturally infected adult cows, with or without clinical signs, were classified histopathologically. Tissue samples obtained focused on gutassociated lymphoid tissue. Lesions were divided into five categories. Focal lesions (n=68 cases), consisted of small granulomas in the ileal and jejunal lymph nodes or the ileocaecal lymphoid tissue. In the multifocal type (n=13 cases), small granulomas or scattered giant cells appeared in some intestinal villi, as well as in the lymph nodes. Diffuse multibacillary lesions (n=15 cases), associated with severe granulomatous enteritis affecting different intestinal locations and lymph nodes, were formed by macrophages containing large numbers of acid-fast bacilli. In diffuse lymphocytic lesions (n=3 cases), lymphocytes were the main inflammatory cells, with some macrophages or giant cells containing few if any mycobacteria. In diffuse intermediate forms (n=17 cases), the infiltrate was formed by abundant lymphocytes and macrophages, and mycobacteria were present to varying degrees related to the number of macrophages. Clinical signs and gross lesions were mainly associated with diffuse forms. Thickening of the intestinal wall, which was the most common macroscopical finding, was related to the degree of submucosal change. Mycobacterium avium subsp. paratuberculosis was identified by culture or PCR in all cows with diffuse lesions, and in 55.5 and 37% of those with multifocal or focal forms, respectively. The importance of sampling the ileal and caudal jejunal lymph nodes to find histological lesions of paratuberculosis in cattle is emphasized.

Author: Yoshioka, K.; Enaga, S.; Taniguchi, K.; Fukushima, U.; Uechi, M.; Mutoh, K. Year: 2004 Title: Morphological characterization of ductular reactions in canine liver disease Journal: J Comp Pathol Volume: 130 Issue: 2-3 Pages: 92-8 Date: Feb-Apr Abstract: Intrahepatic bile duct proliferation (ductular reaction) was examined histologically, immunohistochemically and ultrastructurally in four cases of canine liver disease, diagnosed as chronic hepatitis, liver fibrosis, cirrhosis and cholangiocellular carcinoma. Ductular reaction was a common finding in all cases. Most of the proliferated bile ducts were similar to normal bile ducts. In addition, duct-like structures occurred, consisting of hepatocytes and of intermediate cells that had phenotypic characteristics of both cholangiocytes and hepatocytes. The proliferated bile ducts were immunohistochemically negative for proliferating cell nuclear antigen (PCNA) and stem cell

factor (SCF). The proliferated bile ducts in these four cases of canine liver disease thus showed both typical ductular reactions, such as elongation and tortuosity of the existing bile ducts, and atypical ductular reactions resulting from metaplasia of hepatocytes.

Author: Solano-Gallego, L.; Fernandez-Bellon, H.; Morell, P.; Fondevila, D.; Alberola, J.; Ramis, A.; Ferrer, L. Year: 2004 Title: Histological and immunohistochemical study of clinically normal skin of Leishmania infantum-infected dogs Journal: J Comp Pathol Volume: 130 Issue: 1 Pages: 7-12 Date: Jan Abstract: Skin lesions are the most usual manifestation of canine leishmaniosis. The aim of this study was to investigate the histological pattern and parasite load in clinically normal skin of Leishmania-infected dogs. Two groups of Leishmania-infected dogs were studied. Group A consisted of 15 symptomless animals which, although seronegative or only mildly seropositive, gave a positive polymerase chain reaction (PCR) for Leishmania in the skin. Group B consisted of 20 clinically affected dogs which were highly seropositive and PCR-positive. Biopsies of normal skin from all dogs were processed for routine histology and Leishmania immunohistochemistry. The study demonstrated microscopical lesions and the presence of parasites in the skin from dogs of group B, but not group A. The results cast doubt on the relevance of infected but symptomless dogs in the epidemiology of canine leishmaniosis. In contrast, however, the clinically normal skin of sick dogs harbours the parasite and probably plays a role in the transmission of leishmaniosis.

Circovirus
Author: Opriessnig, T.; Janke, B. H.; Halbur, P. G. Year: 2006 Title: Cardiovascular lesions in pigs naturally or experimentally infected with porcine circovirus type 2 Journal: J Comp Pathol Volume: 134 Issue: 1 Pages: 105-10

Date: Jan Abstract: Abundant intracytoplasmic porcine circovirus type 2 (PCV2) was associated with myocardiocyte swelling or necrosis, or myocardial fibrosis (or both) in three naturally infected pigs aged 4-7 weeks from three different farms. One 6 week old pig from a fourth farm had severe diffuse segmental to circumferential lymphohistiocytic and plasmacytic periarteritis and endarteritis in several organs, PCV2 antigen was demonstrated in endothelial cells, and inflammatory cells in the arterial walls. In three pigs experimentally infected with PCV2, viral antigen was also associated with obliterated blood vessels in areas of granulomatous and necrotizing lymphadenitis. Together these findings suggest that the cardiovascular system in general and endothelial cells in particular play an important role in the pathogenesis of PCV2-associated diseases.

Maedi-Visna
Author: Benavides, J.; Fuertes, M.; Garcia-Pariente, C.; Ferreras, M. C.; Garcia Marin, J. F.; Perez, V. Year: 2006 Title: Natural cases of visna in sheep with myelitis as the sole lesion in the central nervous system Journal: J Comp Pathol Volume: 134 Issue: 2-3 Pages: 219-30 Date: Feb-Apr Abstract: Of 118 sheep with visna, 12 showed myelitis as the only nervous lesion. They were ovine lentivirus (OvLV)-seropositive and provirus DNA was demonstrated by LTR-PCR in all the samples with lesions. Clinically, all showed hindlimb paralysis and some were completely recumbent. Grossly, a swollen and discoloured area was identified in the white matter in 10 sheep. Microscopical changes consisted of a wedge-shaped area of non-suppurative leucomyelitis with mononuclear perivascular cuffing, demyelination and white matter degeneration. Except for two samples, grey matter was affected adjacent to severe white matter lesions. Three different microscopical patterns of lesion were identified, all having in common the presence of perivascular inflammation: the so-called vascular pattern was characterized by perivascular cuffs with minimal lesions in the adjacent neuroparenchyma; the malacic pattern, which was the commonest type, was characterized by severe white matter destruction and small numbers of macrophages; and the infiltrative pattern was characterized by a severe infiltrate of histiocytes in the parenchyma. Maedivisna virus antigen was detected immunohistochemically only in areas with lesions, and the degree of immunolabelling was unrelated to the severity of the damage. Diagnosticians should bear in mind that a considerable number of visna cases show only spinal cord lesions. Examination of paraffin wax-embedded samples by LTR-PCR and immunohistochemistry would seem useful in confirming a histopathological diagnosis of visna from spinal cord samples.

Author: Angelopoulou, K.; Brellou, G. D.; Vlemmas, I. Year: 2006 Title: Detection of maedi-visna virus in the kidneys of naturally infected sheep Journal: J Comp Pathol Volume: 134 Issue: 4

Pages: 329-35 Date: May Abstract: Infections with maedi-visna virus (MVV) cause progressive inflammation in different organs, mainly the lung, mammary gland, brain and joints. The aim of the present study was to investigate whether the kidney represents a viral target in natural MVV infection. For this, kidney samples from 13 sheep naturally infected with MVV were examined by histology, polymerase chain reaction (PCR), and immunohistochemistry. The kidneys of nine animals showed membranoproliferative glomerulonephritis and interstitial nephritis. The inflammatory infiltrate consisted of lymphocytes, plasma cells and macrophages. Interestingly, lymphoid follicles resembling those known to occur in other MVV-infected tissues were observed. Lung tissue from the same animals had typical MVV lesions, such as lymphofollicular hyperplasia and interstitial pneumonia. Maedi-visna proviral DNA sequences were detected in renal and lung tissue samples from these nine sheep by PCR, and the specificity of the amplified products was further verified by DNA sequencing. Moreover, MVV-specific immunohistochemistry revealed viral antigen in affected kidneys and lungs. These results suggest that the kidney may be a common target in natural MVV infection, and raise the issue of the role of this organ in the disease.

Author: Biescas, E.; Preziuso, S.; Bulgin, M.; DeMartini, J. C. Year: 2005 Title: Ovine lentivirus-associated leucomyelitis in naturally infected North American sheep Journal: J Comp Pathol Volume: 132 Issue: 2-3 Pages: 107-16 Date: Feb-Apr Abstract: Leucomyelitis was the predominant feature in four North American adult sheep (cases 14) with ovine lentivirus (OvLV) infection. All four animals were OvLV-seropositive and a syncytogenic virus consistent with OvLV was isolated from the brain of case 3 and the lungs of case 4. Clinically, the sheep had dyspnoea and neurologic signs of varying severity. Changes in the central nervous system included asymmetrical meningoleucomyelitis with white matter degeneration in all four sheep and scattered foci of leucoencephalitis in periventricular, subependymal and other white matter areas of the brain of the three animals (cases 1, 2 and 4) for which the brain was examined. In the lungs of two sheep (cases 3 and 4), there was lymphoid interstitial pneumonia with marked lymphoid hyperplasia. The viral capsid antigen (p25) was detected by immunohistochemistry (IHC) in sections of lung, brain and spinal cord of the four sheep and OvLV RNA was detected by in-situ hybridization (ISH) in lung and spinal cord samples. The results confirm the usefulness of the IHC and ISH for differential diagnosis of visna.

Scrapie
Author: Jeffrey, M.; Gonzalez, L.; Chong, A.; Foster, J.; Goldmann, W.; Hunter, N.; Martin, S. Year: 2006 Title: Ovine infection with the agents of scrapie (CH1641 isolate) and bovine spongiform encephalopathy: immunochemical similarities can be resolved by immunohistochemistry Journal: J Comp Pathol Volume: 134 Issue: 1 Pages: 17-29 Date: Jan

Abstract: Immunochemical ("rapid") tests, which recognize a partly protease-resistant conformer of the prion protein (PrP(res)) are now widely used in Europe for the diagnosis of transmissible spongiform encephalopathies (TSEs). Some of these tests can be used to distinguish natural scrapie from experimental bovine spongiform encephalopathy (BSE) in sheep, on the basis of migration pattern differences of PrP(res) in Western immunoblots. However, PrP(res) from sheep inoculated with CH1641 scrapie gives an immunoblot profile similar to that of sheep inoculated with BSE. Therefore, field scrapie strains similar to CH1641 might be misclassified as ovine BSE in the rapid tests currently employed. This study confirmed that the Western blot similarities (size of the unglycosylated band and distinct reactivity with 6H4 and P4 antibodies) between CH1641 and BSE remained consistent regardless of the PrP genotype of the sheep, but the two infections resulted in accumulation of disease-associated PrP (PrP(d)) that could easily be distinguished by the immunohistochemical "peptide mapping" method. This method, which reveals conformational differences of PrP(d) by the use of a panel of antibodies, indicated that PrP(d) from the CH1641 isolate was truncated further upstream in the N terminus than was PrP(d) from other ovine TSEs, including experimental BSE. In addition, the immunohistochemical "PrP(d) profile method", which defines the phenotype of PrP(d) accumulation in the brain of affected sheep, showed that CH1641 infection leads to much more intra-neuronal and considerably less extracellular PrP(d) than does experimental BSE. The overall results demonstrate that a combined Western blotting and immunohistochemical approach is required to discriminate between different TSE strains in sheep.

Author: Jeffrey, M.; Martin, S.; Gonzalez, L.; Foster, J.; Langeveld, J. P.; van Zijderveld, F. G.; Grassi, J.; Hunter, N. Year: 2006 Title: Immunohistochemical features of PrP(d) accumulation in natural and experimental goat transmissible spongiform encephalopathies Journal: J Comp Pathol Volume: 134 Issue: 2-3 Pages: 171-81 Date: Feb-Apr Abstract: Scrapie is a transmissible spongiform encephalopathy (TSE) or prion disease, which naturally affects sheep and goats. Immunohistochemical epitope mapping of abnormal PrP accumulations (PrP(d)) in brain can help in characterizing sheep TSE sources or strains and in identifying potential bovine spongiform encephalopathy (BSE) infections of sheep. Natural and experimental TSE infections of goats were examined to determine whether the epitope mapping approach could also be applied to aid recognition of BSE infection in goats. Goats experimentally infected with the SSBP/1 or CH1641 sheep scrapie strains or with cattle BSE, together with four field cases of natural TSE in goats, were examined immunohistochemically with six different antibodies. CH1641 and SSBP/1 infections in goats, as in sheep, showed PrP(d) accumulations which were mainly intracellular. Some differences in targeting, particularly of Purkinje cells, was evident in inter-species comparisons of CH1641 and SSBP/1. PrP(d) labelling of goat BSE experimental cases showed extensive intracellular and extracellular accumulations, also similar to those in sheep BSE. Intra-neuronal PrP(d) in both goat and sheep BSE was labelled only by antibodies recognizing epitopes located C-terminally of residue His99, whereas in natural sheep TSE sources, and in sheep and goat SSBP/1, PrP(d) was also detected by antibodies to epitopes located between residues Trp93 and His99. Testing of four natural goat TSE samples showed one case in which epitope mapping characteristics and the overall patterns of PrP(d) accumulation was identical with those of experimental goat BSE. The four natural goat scrapie cases examined showed some degree of immunohistochemical phenotype variability, suggesting that multiple strains exist within the relatively small UK goat population.

Author: Thuring, C. M.; van Keulen, L. J.; Langeveld, J. P.; Vromans, M. E.; van Zijderveld, F. G.; Sweeney, T. Year: 2005 Title: Immunohistochemical distinction between preclinical bovine spongiform encephalopathy and scrapie infection in sheep Journal: J Comp Pathol Volume: 132 Issue: 1 Pages: 59-69 Date: Jan Abstract: Sheep are susceptible experimentally to bovine spongiform encephalopathy (BSE), the clinical signs being indistinguishable from those of scrapie. Because of the possibility of natural ovine BSE infection, laboratory tests are needed to distinguish between scrapie and BSE infection. The objectives of this study were to determine whether (1) PrPSc accumulates in biopsy samples of the tonsil or third eyelid, or both, of BSE-infected sheep before the appearance of clinical disease, and (2) such samples from BSE- and scrapie-infected sheep differ in respect of PrPSc accumulations. Homozygous ARQ sheep (n = 10) were dosed orally at 4-5 months of age with a brain homogenate from BSE-infected cattle. Third eyelid and tonsillar biopsy samples were taken at < or = 6 monthly intervals post-infection and examined immunohistochemically for PrPSc. Third eyelid protuberances were difficult to identify, resulting in many unsuitable samples; however, third eyelid samples shown to contain lymphoid follicles were invariably negative for PrPSc. In contrast, tonsillar biopsy samples became positive for PrPSc from 11 to 20 months post-infection. Consistent differences in the morphology of PrPSc granules in tingible body macrophages (TBMs) between BSE- and scrapie-infected sheep were detected with anti-peptide antibodies directed towards amino acids 93-106 of the ovine prion protein: thus, PrPSc appeared as single granules in TBMs of tonsillar sections from BSE-infected sheep, whereas clusters of PrPSc granules were observed within TBMs in the tonsils of scrapie-infected sheep. In contrast, antibodies against epitopes situated N- and C-terminally from the 93-106 region of the ovine prion protein revealed no differences between BSE- and scrapie-infected sheep in terms of PrPSc granules in TBMs.

Author: Green, R.; Horrocks, C.; Wilkinson, A.; Hawkins, S. A.; Ryder, S. J. Year: 2005 Title: Primary isolation of the bovine spongiform encephalopathy agent in mice: agent definition based on a review of 150 transmissions Journal: J Comp Pathol Volume: 132 Issue: 2-3 Pages: 117-31 Date: Feb-Apr Abstract: In the epizootic of bovine spongiform encephalopathy (BSE) in Great Britain, the cattle in which a positive diagnosis was made numbered almost 180 000, but strain characterization was performed on only a very small sample of these cases. This report describes the results of BSE transmission to Prnp(a) mice from 150 transmission experiments at the Veterinary Laboratories Agency (VLA) over the last decade. These data, derived from a large sample of BSE-affected cattle, confirmed previous reports that show no evidence for diversity in BSE isolates. The agent was readily transmitted to mice, with a mean incubation period of 408 days in the RIII strain. Because the incubation period was related to the titre of the inoculum, it is not a reliable characteristic of strain type on primary isolation. Consistent neuropathological changes associated with infection by the BSE agent in RIII and C57Bl mice included focal vacuolation in the dorsal

cochlear nuclei, vacuolation of the granule cell layer of the cerebellum, absence of lesions in the hippocampus and in the molecular layer of the cerebellum, and a fine particulate distribution of disease-specific PrP (demonstrated immunohistochemically), with few or no amyloid plaques. These features, together with the conventional lesion profile, will be of use in distinguishing the agents of BSE and scrapie in sheep.

Author: Ligios, C.; Dexter, G.; Spiropoulos, J.; Maestrale, C.; Carta, A.; Simmons, M. M. Year: 2004 Title: Distribution of vascular amyloid in scrapie-affected sheep with different genotypes Journal: J Comp Pathol Volume: 131 Issue: 4 Pages: 271-6 Date: Nov Abstract: Vascular amyloidosis in the brain is a pathological feature of ovine scrapie. Its occurrence varies between sheep, but whether this variation reflects differences in the host or the infecting scrapie strain (or both) is not clear. To investigate whether amyloidosis, like vacuolation and PrPsc distribution, is associated with genotype, the brains from 131 sheep representing a range of genotypes commonly associated with scrapie were examined histologically and immunohistochemically. Vascular amyloidosis was absent in 66 sheep, 59 of which were of the ARQ/ARQ genotype and seven the ARQ/AHQ genotype. In contrast, it was found in four of 39 ARQ/VRQ sheep (10.2%) and in 10 of 26 VRQ/VRQ sheep (38.4%). The distribution of amyloid was highly variable, but the most severely affected areas were the lateral geniculate nuclei (five cases) and the ventral thalamic nuclei (four cases). No amyloidosis was found in the medulla or in the basal nuclei. From this preliminary study it was concluded that amyloidosis is relatively rare in sheep with scrapie. Moreover, its occurrence appeared to depend on the presence of at least one valine at codon 136.

PRRS

Author: Cheon, D. S.; Chae, C. Year: 2004 Title: Comparison of the pathogenicity of two strains (wild type and vaccine-like) of porcine reproductive and respiratory syndrome virus (PRRSV) in experimentally infected sows Journal: J Comp Pathol Volume: 130 Issue: 2-3 Pages: 105-11 Date: Feb-Apr Abstract: The objective of this study was to compare two Korean strains of porcine reproductive and respiratory syndrome virus (PRRSV), namely a wild type (WT) strain and a vaccine-like (VL) strain, in respect of pathogenicity and viral distribution in the tissues. Both strains were of the North American genotype. Two groups of five pregnant sows were infected with either the WT or the VL strain 2 weeks before their expected farrowing date. The WT strain-inoculated sows showed abortion and premature farrowing, whereas the VL strain-inoculated sows remained clinically normal and did not farrow prematurely. Of the 18 liveborn piglets from the WT strain-inoculated sows, 14 had interstitial pneumonia. Of the 60 liveborn piglets from the VL strain-inoculated sows, only six had interstitial pneumonia. PRRSV antigen or nucleic acid was detected in 48/65 (73.8%)

of stillborn and liveborn piglets from the WT strain-inoculated sows, but in only 12/64 (18.8%) of stillborn and liveborn piglets from the VL strain-inoculated sows.

Distemper
Author: Friess, M.; Engelhardt, P.; Dobbelaere, D.; Zurbriggen, A.; Grone, A. Year: 2005 Title: Reduced nuclear translocation of nuclear factor (NF)-kappaB p65 in the footpad epidermis of dogs infected with distemper virus Journal: J Comp Pathol Volume: 132 Issue: 1 Pages: 82-9 Date: Jan Abstract: Infection of canine footpads with the canine distemper virus (CDV) can cause massive epidermal thickening (hard pad disease), as a consequence of increased proliferation of keratinocytes and hyperkeratosis. Keratinocytes of canine footpad epidermis containing detectable CDV nucleoprotein antigen and CDV mRNA were shown previously to have increased proliferation indices. Because various proteins that play a role in the proliferation of epidermal cells are viral targets, the potential participation of such proteins in CDV-associated keratinocyte proliferation was investigated. Transforming growth factor-alpha (TGF-alpha), cell cycle regulatory proteins p21, p27 and p53, and nuclear factor (NF)-kappaB transcription factor components p50 and p65 were studied in the footpad epidermis from the following groups of dogs inoculated with CDV: group 1, consisting of seven dogs with clinical distemper and CDV in the footpad epidermis; group 2, consisting of four dogs with clinical distemper but no CDV in the footpad epidermis; group 3, consisting of eight dogs with neither clinical distemper nor CDV in the footpad epithelium. Group 4 consisted of two uninoculated control dogs. The expression of TGF-alpha, p21, p27 and p53, and p50 in the basal layer, lower and upper spinous layers, and in the granular layer did not differ statistically between CDV-positive (group 1) and CDV-negative (groups 2-4) footpad epidermis. However, there were differences in the levels of nuclear and cytoplasmic p65 expression between group 1 dogs and the other three groups. Thus, footpads from group 1 dogs had more keratinocytes containing p65 in the cytoplasm and, conversely, fewer nuclei that were positive for p65. These findings indicate that p65 translocation into the nucleus is reduced in CDV-infected footpad epidermis. Such decreased translocation of p65 may help to explain increased keratinocyte proliferation in hard pad disease and suggests interference of CDV with the NFkappaB pathway.

Foot-and-Mouth

Author: Quan, M.; Murphy, C. M.; Zhang, Z.; Alexandersen, S. Year: 2004 Title: Determinants of early foot-and-mouth disease virus dynamics in pigs Journal: J Comp Pathol Volume: 131 Issue: 4 Pages: 294-307 Date: Nov

Abstract: This paper provides a quantitative description of the early infectious process in pigs experimentally infected with foot-and-mouth disease virus (FMDV), obtained by dose-dependent, time course studies of viral load in serum. Pigs were inoculated by the intravenous or intradermal/subcutaneous route with FMDV and housed together in groups or individually. The effects of dose, inoculation route and exposure intensity on the replication of FMDV in vivo and the development of disease were studied. It was shown that the higher the dose, the shorter was the time to the start of active viraemia and to the onset of clinical signs. Exposure intensity and housing conditions influenced the viral dynamics of FMDV. Increasing the exposure intensity, by increasing the number of infected pigs housed together, had the effect of synchronizing the infection and reducing the variance in the start of active viraemia. Increasing the number of pigs housed together also increased the interaction between the pigs and the activity of individual pigs, which had the effect of shortening the time to the onset of clinical signs such as vesicle formation. Intradermal inoculation was more effective than intravenous inoculation for transmitting FMDV to pigs, resulting in shorter times to the start of active viraemia and in higher clinical scores.

Sign up to vote on this title
UsefulNot useful