Enzootic Haematuria Research Articles

Pathological study of non-neoplastic urinary bladder lesions in cattle and buffaloes: a preliminary report

A total of 236 urinary bladders (94 cattle and 142 buffaloes) collected from Bareilly, Uttar Pradesh, India, were studied for spontaneous lesions. These adult animals belonged to Institute's organized dairy farm and rural areas in the Rohilkhand region of Uttar Pradesh. Grossly, congestion, hemorrhages, and cystoliths in urinary bladders were diagnosed. Histopathologically, the major conditions diagnosed were acute cystitis, 44 (18.64%), including, congestion, hemorrhages, sub-acute cystitis; chronic cystitis, 74 (31.35%), including chronic cystitis un-complicated type, lymphocytic cystitis, plasmolymphocytic cystitis, follicular cystitis, hyperplasia, nodular/acinar hyperplasia, and cystolithiasis; and nothing unusual diagnosed, 118 (50.00%). Similar types of pathological conditions were diagnosed in both species of animals with exception of follicular cystitis and nodular/acinar hyperplasia which was diagnosed respectively only in buffaloes and cystoliths in cows. In addition, a good number of 17/25 (68%) urinary bladder samples tested were found positive for presence of bovine papillomavirus type-2 (BPV-2) by polymerase chain reaction. These included eight cases of acute cystitis, an equal number of cases of chronic cystitis, and one normal bladder. BPV-2 is known as potential source of enzootic bovine hematuria along with other co-factors in enzootic areas. Lesions of zoonotic significance, like tuberculosis, etc., were not diagnosed. None of the observed lesions represented conditions, which, by themselves, would warrant carcass condemnation in buffaloes.

Clotting profile in cattle showing chronic enzootic haematuria (CEH) and bladder neoplasms

Primary haemostasis (bleeding and blood clotting time), activated partial thromboplastin time (APTT), prothrombin time (PT), antithrombin III (ATIII), protein C, protein S, fibrinogen and D-dimer were determined in 13 cattle affected by chronic enzootic haematuria (CEH) and bladder neoplasms and 10 healthy cattle (control group). Increases in antithrombin III and protein S activities (P<0.01) and protein C and fibrinogen plasma levels (P<0.05) were observed in sick animals, while activated partial thromboplastin time, prothrombin time, and D-dimer did not show significant differences when compared to healthy animals. The clotting profile observed does not seem responsible for the chronic bleeding typical of CEH. The observed modification of some coagulation markers may derive from multiple interactions among cancer, inflammation and viral infection status typical of this syndrome.

Preliminary assessment of binary ethylenimine inactivated and saponized cutaneous warts (BPV-2) therapeutic vaccine for enzootic bovine haematuria in hill cows

A preliminary therapeutic vaccine trial was conducted in hill cows to evaluate the therapeutic potential of binary ethylenimine (BEI) inactivated and saponized bovine papillomavirus-2 (BPV-2) for enzootic bovine haematuria (EBH). Although the vaccine failed to show favorable clinical vaccine results in treatment of EBH affected cows at 120 days post-vaccination but immunopathological responses were encouraging. A significant difference was observed in humoral (against Brucella abortus strain 19S) and cell-mediated (in vivo phytohaemagglutination delayed type hypersensitivity (PHA DTH) test and CD4+/CD8+ T-cells ratio by FACS analysis) immune responses following vaccination. The vaccinated animals grossly failed to show regression of bladder tumours but microscopically engorgement and marked perivascular infiltration of mononuclear cells was observed which are indicative of the induction of initial stages of tumour regression. Overall results indicated that the therapeutic vaccine developed can have potentials for treating EBH in cows, for which further modifications in vaccine dose and field trial is required.

Detection and Quantification of Bovine Papilloma Virus Type 2 (BPV-2) by Real-time PCR in Urine and Urinary Bladder Lesions in Enzootic Bovine Haematuria (EBH)-Affected Cows

This study was conducted with the objectives of detecting bovine papillomavirus type 2 (BPV-2) in urine samples and urinary bladder lesions in bovines using polymerase chain reaction (PCR) and real-time PCR-based molecular diagnostic tests, and quantifying BPV-2 in urinary bladder lesions especially in enzootic bovine haematuria (EBH)-affected animals. BPV-2 viral DNA was detected in urine samples (50%) and urinary bladder tissue (68.6%). Cloning and sequencing results showed a close homology with other Indian BPV-2 sequences. Quantitative real-time PCR (SYBR Green assay) showed that the BPV-2 load was low and similar irrespective of inflammatory or neoplastic lesions in the bladder. It was concluded that BPV-2 DNA is frequently present in urine and urinary bladder lesions in cows in an EBH endemic region and virus load was low in urinary bladder lesions.

Papillomatosis in Buffaloes: A Less-Known Disease

Scant information is available on papillomatosis in buffaloes, and it is an almost unknown disease. It has been described from India, Italy and Turkey. Buffalo papillomatosis occurs in cutaneous and mucosal forms. Cutaneous papillomatosis is manifested as cutaneous wart (CW) and teat papilloma types. The condition is known to be caused by bovine papillomaviruses (BPV)-1 and 2 and their mixed infections. Buffalo CWs are experimentally transmissible to hamsters, cattle as well as buffaloes. Once BPV establishes infection in buffaloes, infection spreads from buffalo to buffalo, without cattle intermediary. Histologically, CWs are mostly diagnosed as fibropapillomas. The mucosal form occurs as urinary bladder tumours similar to enzootic bovine haematuria which is also associated with bracken fern infested areas. BPVs are yet to be demonstrated in teat papillomas and urinary bladder tumours of buffalo cases. Papillomatosis in buffaloes is a little-known disease, but it is a separate infectious ailment of buffaloes and deserves more attention by researchers.

A Retrospective Study of 586 Tumours in Brazilian Cattle

Records from 6,706 necropsy examinations of cattle performed over a 45-year period were surveyed and 586 cases of neoplasia were identified. The organ system most frequently affected by neoplastic disease (n=139 cases) was the alimentary tract. This finding was attributed to a high incidence of squamous cell carcinoma of the upper alimentary tract associated with the chronic ingestion of bracken fern (Pteridium aquilinum). This carcinogenic plant was also associated with a relatively high incidence (n=35 cases) of urinary bladder tumours (enzootic haematuria). Tumours of the alimentary tract were followed, in decreasing order of frequency, by tumours of the skin and subcutis (n=129 cases), haemopoietic tissue (n=101 cases), the eye and periorbital tissues (n=88 cases), the urinary system (n=44 cases), the female reproductive system (n=21 cases), the endocrine system (n=16 cases), the liver and pancreas (n=12 cases), the nervous system (n=6 cases), the respiratory system (n=6 cases) and the mammary gland (n=1 case). The primary anatomical location of 16 tumours was undetermined.

Investigations On The Pathology of Enzootic Bovine Haematuria in Turkey

ummary 1Bovine haematuria occurs in Turkey among cattle and buffaloes. The disease manifests itself in bleeding in the bladder. The bladder is the primary seat of the affection. 2The pathological changes consisted of neoplastic proliferation of both the epithelial and mesenchymal tissue of the bladder, up to the most malignant forms of new growth. 3Papilloma and papillary carcinoma were commoner than infiltrative carcinoma. Malignant epithelial tumours occurred more frequently than benign ones. The proportion of malignant epithelial tumours to benign ones was 1.4:1. Epithelial tumours were more common in males than in females. 4Mesoblastic tumours of the bladder consisted of haemangioma and sarcoma. Haemangioma was the commonest finding. Usually epithelial and mesoblastic tumours occurred simultaneously. Tumours of the bladder were the actual cause of the vesical haematuria. Zusammenfassung Untersuchungen uber die Pathologie der enzootischen Rinderhamaturie in der Turkei 1Die Rinderhamaturie kommt in der Turkei sowohl bei Rindern als auch bei Buffeln vor. Es treten Blutungen in der Harnblase, dem primaren Sitz der Affektion, auf. 2Die pathologischen Veranderungen bestehen in Proliferationen des epithelialen und mesenchymalen Gewebes der Harnblase in teilweise sehr bosartiger Form. 3Papillome und papillare Karzinome wurden haufiger als infiltrierende Karzinome beobachtet, bosartige epitheliale Tumoren in starkerem Mase als gutartige. Das Verhaltnis der bosartigen epithelialen Tumoren zu den gutartigen war wie 1,4:1. Epitheliale Tumoren wurden haufiger bei mannlichen als bei weiblichen Tieren beobachtet. 4Die mesoblastischen Tumoren der Harnblase bestanden in Hamangiomen und Sarkomen. Gewohnlich wurden bei der Hamaturie der Rinder Hamangiome nachgewiesen, epitheliale und mesoblastische Tumoren traten gleichzeitig auf. Als Ursache der vesikalen Hamaturie sind Tumoren der Harnblase anzusehen. Resume Recherches anatomo-pathologiques sur l'hematurie enzootique en Turquie 1L'hematurie des bovides se rencontre en Turquie aussi bien chez le boeuf que chez le buffle. Elle est provoquee par des hemorrhagies de la vessie, localisation primaire de l'affection. 2Les lesions anatomo-pathologiques consistent en proliferations des tissus epitheliaux et conjonctifs, sous une forme parfois tres maligne. 3Des papillomes et des carcinomes papillomateux furent observes plus souvent que des carcinomes infiltratifs et des tumeurs epitheliales malignes plus frequemment que les benignes. Le rapport entre le nombre des tumeurs epitheliales malignes et le nombre des tumeurs epitheliales benignes etait de 1,4: 1. Les tumeurs epitheliales se rencontrent plus souvent chez les animaux mâles que chez les animaux femelles. 4Les tumeurs du tissu conjonctif de la vessie comprenaient des hemangiomes et des sarcomes. En general, on put deceler des hemangiomes dans l'hematurie des bovides. Les tumeurs epitheliales et conjonctives etaient presentes en meme temps. La cause de l'hematurie vesicale doit etre cherchee dans la presence de tumeurs de la vessie. Resumen Investigaciones sobre la patologia de la hematuria enzootica bovina en Turquia 1La hematuria bovina se da en Turquia tanto en los bovidos como tambien en los bufalos. En la vejiga urinaria aparecen hemorragias, siendo la misma el lugar del asentamiento primario de la afeccion. 2Las alteraciones patologicas consistent en proliferaciones de los tejidos epitelial y mesenquimatoso de la vejiga de la orina, en parte de una forma bastante maligna. 3Los papilomas y los carcinomas papilomatosos se observaron con una frecuencia mayor que los carcinomas infiltrantes, y los tumores malignos epiteliales en mayor grado que los benignos. La relacion entre los tumores epiteliales malignos y los benignos era como 1,4: 1. Los tumores epiteliales se observaron con mayor frecuencia en los animales masculinos que en las hembras. 4Los tumores mesoblasticos de la vejiga de la orina consistian en hemangiomas y sarcomas. Por lo general, en la hematuria de los bovidos se identificaron hemangiomas, apareciendo al mismo tiempo unos tumores epiteliales y mesoblasticos. Los tumores de la vejiga urinaria se han de considerar como la etiologia de la hematuria vesical.

Serum mucoprotein and sialic acid in enzootic bovine haematuria.

Summary Estimations of serum mucoprotein and sialic acid were carried out at fortnightly intervals up to the 60th day in four healthy and seven haematuric hill cattle 2–4 years old. There was a significant increase in the level of both these constituents throughout the period of study in animals with chronic bovine haematuria. This is ascribed to the neoplastic growths in the urinary bladder in this condition. Zusammenfassung Sialsauregehalt im Blutserum von Kuhen mit enzootischer Hamaturie Bei 4 gesunden und 7 an Hamaturie-leidenden Kuhen der Gebirgsrasse (hill cattle) im Alter von 2–4 Jahren wurden wahrend 60 Tagen vierzehntaglich das Mucoprotein und die Sialsaure im Blutserum bestimmt. Die beiden Substanzen zeigten bei den an Hamaturie erkrankten Tieren eine signifikante Konzentrationserhohung gegenuber den gesunden Kuhen. Diese Veranderung wird dem neoplastischen Charakter der Harnblasenerkrankung zugeschrieben. Resumen Mucoproteinas y acido sialico sericas en la hematuria bovina enzootica En 4 vacas sanas y 7 enfermas de hematuria, pertenecientes todas ellas a la raza montanesa india, de edades comprendidas entre 2–4 anos, se valoraron durante 60 dias cada catorce dias las mucoproteinas y el acido sialico en el suero sanguineo. Ambas substancias mostraban en los animales enfermos de hematuria un aumento significante en las concentraciones frente a las de las vacas sanas. Esta modificacion se atribuye al caracter neoplasico en la vejiga de la orina en estas condiciones.

Association of bovine papillomavirus type 2 (BPV-2) and urinary bladder tumours in cattle from the Azores archipelago

Urinary bladder tumours in cattle are caused by chronic ingestion of bracken fern and BPV-1/2 infection. The objective of the present study was to assess if BPV-2 was present in urinary bladder lesions from cattle with chronic enzootic haematuria (CEH) from the Azores archipelago (Portugal), in order to gain further information regarding the epidemiologic distribution of this virus. Samples were analysed using PCR specific primers for BPV-2 DNA and an immunohistochemistry for BPV E5 oncoprotein detection. We found a 28% incidence rate of BPV-2 DNA in different types of tumours and cystitis cases (13 out of 46 samples). Tested positive samples for PCR were also positive for the viral E5 oncoprotein; protein immunolabeling was mainly detected within the cytoplasm of urothelial cells, displaying a juxtanuclear distribution. This is the first report of BPV-2 detection in urinary bladder tumours associated with CEH in cattle from the Azores archipelago.

Epithelioid Hemangiosarcomas of the Bovine Urinary Bladder: A Histologic, Immunohistochemical, and Ultrastructural Examination of Four Tumors

Epithelioid hemangiosarcoma is a specific variant of hemangiosarcoma that has recently been recognized in domestic animals. These malignant vascular neoplasms histologically resemble, and may be mistaken for, carcinomas. Four epithelioid hemangiosarcomas in the urinary bladders of 4 cows with severe enzootic hematuria are described in the current study. Grossly, the vesicular mucosa of the urinary bladder of each cow contained a single red elevated nodule. Histologically, each neoplasm was composed of short strands, cords, or nests of epithelioid, round, or slightly spindle-shaped endothelial cells that formed small vascular structures. Neoplastic cells were immunohistochemically positive for factor VIII-related antigen and vimentin, and were negative for cytokeratin and desmin. Ultrastructurally, the neoplastic cells often contained cytoplasmic intermediate filaments, a prominent granular endoplasmic reticulum, a Golgi complex, mitochondria, marked pinocytotic activity, and rare Weibel-Palade bodies. These neoplasms were diagnosed as epithelioid hemangiosarcomas based on their histologic, immunohistochemical, and ultrastructural features. The present report widens the spectrum of mesenchymal tumors of the bovine urinary bladder and aids in the characterization of these vascular neoplasms.

INTOXICAÇÃO AGUDA POR PTERIDIUM AQUILINUM EM DOIS BOVINOS

TWO CASES OF ACUTE POISONING BY PTERIDIUM AQUILINUM IN CATTLE Pteridium aquilinum, also known as bracken fern, is a cosmopolitan plant that grows better in cold areas of moderate rainfall rate with low-fertility acid sandy soils. All parts of the plant contain active toxic principles. Some of the factors that lead to the intoxication of animals are hunger, lack of fibrous food, ingestion of contaminated hay, and the fact that the plant can be addictive. Considering the daily amount ingested and the period of ingestion, cattle can present three different clinical manifestations: 1) Acute hemorrhagic diathesis; 2) Bladder neoplasia or enzootic hematuria; 3) Digestive tract neoplasia. Two animals were attended to at the Veterinary Hospital of UNESP, Botucatu, presenting signs of acute poisoning by bracken fern. The diagnosis was based on anamnesis, clinical signs, complementary exams, and necropsy findings, which revealed hemorrhages and petechiae in the mucous membranes, pancytopenia, hemorrhages in the mucous and serous membranes, and aplasia of the bone marrow, respectively.

ACHADOS CLÍNICOS, LABORATORIAIS, ULTRASSONOGRÁFICOS E ANÁTOMO-PATOLÓGICOS EM UM CASO DE HEMATÚRIA ENZOÓTICA BOVINA

Pteridium aquilinum is a well-known toxic plant worldwide, responsible for three different clinical syndromes in cattle, including bovine enzootic hematuria (BEH). The present work describes a case study of BEH through clinical, laboratory, ultrasonographic, and pathological findings. A 10-year-old crossbreed was clinically examined. Pale mucous, progressive weight loss, tachycardia, ruminal hypomotility, hematuria, and dysuria were evidenced. Rectal ultrasonography showed a thickened urinary bladder wall, and hyperecoic areas in the lumen suggesting blood clots. Hematology revealed normocytic and hypochromic anemia, hypoproteinemia, and inversion in the lymphocyte/neutrophile ratio. Severe hematuria and proteinuria were observed in urinalysis. Escherichia coli was isolated from the urine culture in a microbiological analysis. Pathological findings in the bladder consisted of submucosa thickness, angiogenesis, and lesions typical of hemangiomas. Ultrasonography may be a helpful tool for providing epidemiological data and clinical signs in the diagnosis of BEH. However, histopathology remains the most reliable method to confirm toxicosis.

Aspectos clínico-hematológicos e lesões vesicais na intoxicação crônica espontânea por Pteridium aquilinum em bovinos

Foram estudados casos espontâneos de intoxicação crônica por samambaia (Pteridium aquilinum) em bovinos nas formas clinicopatológicas de carcinoma de células escamosas (CCE) no trato alimentar superior (TAS) e de hematúria enzoótica bovina (HEB), provenientes da Mesorregião Centro Ocidental Rio-Grandense e encaminhados ao Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria. Para o estudo clínico foram avaliados os sinais clínicos de bovinos com CCEs no TAS e com HEB e realizados hemogramas na fase terminal da doença. Os principais sinais clínicos nos bovinos com CCEs no TAS foram emagrecimento progressivo, atonia ruminal, tosse, disfagia, timpanismo e regurgitação. Nos bovinos com HEB, hematúria foi o principal sinal, observado em todos os casos, seguido de emagrecimento progressivo. No exame hematológico, 33,33% dos bovinos com CCEs no TAS e 66,67% dos bovinos com HEB apresentaram anemia arregenerativa. Alterações no leucograma ocorreram em alguns casos, mas linfopenia foi um achado infreqüente em ambas as formas de intoxicação. Para o estudo morfológico, foram avaliadas as bexigas de 46 bovinos com CCEs no TAS e de 11 bovinos com HEB. Macroscopicamente, 16/46 bexigas dos casos de CCEs no TAS apresentaram alterações macroscópicas, que consistiam nódulos vermelhos ou pálidos, hemorragia e papilomas; urina vermelha foi observada na necropsia de apenas três casos). Nos casos de HEB, os achados macroscópicos vesicais foram nódulos vermelhos, massas neoplásicas focalmente extensas, urina vermelha, papilomas, hemorragias e ruptura de bexiga; pielonefrite e hidronefrose foram observados em poucos casos. Histologicamente, 44/46 (95,65%) bexigas de bovinos com CCEs no TAS apresentaram 22 tipos diferentes de alterações morfológicas, que foram classificadas em alterações neoplásicas (5/22) e alterações não-neoplásicas (17/22); essas últimas foram divididas em alterações epiteliais não-neoplásicas (6/17), alterações gerais na lâmina própria (6/22) e alterações inflamatórias (5/17). Os achados histológicos das bexigas dos casos de HEB foram classificados da mesma forma, resultando em 19 tipos diferentes de alterações morfológicas. Dessas, 5/19 eram alterações neoplásicas e 14/20, alterações não-neoplásicas (9/14 alterações epiteliais não neoplásicas, 3/14 alterações gerais na lâmina própria e 2/14 alterações inflamatórias). Na HEB, os neoplasmas mesenquimais foram mais freqüentemente observados que os epiteliais, e a maior parte era maligna. A técnica de imuno-histoquímica foi utilizada para caracterizar os aspectos morfológicos, principalmente dos neoplasmas. Através do estudo morfológico concluiu-se que é muito freqüente a ocorrência de lesões vesicais em bovinos com a forma crônica de CCEs no TAS e que essas lesões são idênticas às encontradas nos bovinos com HEB.

Prevalência de hematúria enzoótica bovina em rebanhos leiteiros na microrregião do Caparaó, Sul do Espírito Santo, entre 2007 e 2008

A hematúria enzoótica bovina (HEB) é uma doença crônica que tem causado grandes perdas econômicas aos proprietários da microrregião do Caparaó, na região Sul do Estado do Espírito Santo (ES). O objetivo deste trabalho foi obter a prevalência e os achados hematológicos de bovinos com hematúria enzoótica decorrente de intoxicação natural por Pteridium aquilinum (samambaia) no período de março de 2007 a abril de 2008, na microrregião do Caparaó, ES. Foram visitadas 50 propriedades onde foi constatada a presença da planta em todos os 10 municípios avaliados. Cento e oitenta e um bovinos foram examinados e 39 estavam doentes. Dentre os afetados, 22 (56,4%) apresentavam HEB, todos provenientes dos Municípios de Guaçuí, Divino de São Lourenço, Iúna, Ibatiba e Ibitirama. Os bovinos tinham macrohematúria, e os valores do hematócrito mostraram-se baixos, variando de 8% a 31%. A leucometria total teve valor médio de 132 x 10³ mm-3, revelando leucocitose. A temperatura e o tempo de preenchimento capilar (TPC) estavam dentro da normalidade. Quatro bovinos (18,2%) tinham mucosas anêmicas. Nota-se que a prevalência de HEB, na microrregião do Caparaó, ES, é alta e que há a necessidade de novos estudos para diminuir as perdas econômicas causadas por essa doença.

Immunohistochemical Evaluation of Vascular Urinary Bladder Tumors from Cows with Enzootic Hematuria

Fifty-six endothelial-derived urinary bladder tumor samples collected from 26 animals with bovine enzootic hematuria were selected for immunohistochemical studies. Expression of factor VIII-related antigen (FVIIIra), CD31, muscle-specific actin, uroplakin III (UPIII), and the cell cycle-related proteins cyclin D1 and p53 was evaluated in hemangiomas, "hemangioendotheliomas" (a vascular tumor that histologically is intermediate in appearance between a hemangioma and a conventional hemangiosarcoma), and hemangiosarcomas. Although CD31 expression was seen in all endothelial tumors tested, FVIIIra was not expressed in poorly differentiated endothelial tumor cells from solid areas or in 7 muscle-invasive hemangiosarcomas. Cyclin D1 overexpression was seen in 53% of hemangiomas, 82% of hemangioendotheliomas, and 95% of hemangiosarcomas. P53 immunoreactivity was only seen in muscle-invasive hemangiosarcomas. The UPIII staining pattern, normally very intense on the apical aspect and cytoplasm of superficial urothelial cells, was altered in the urothelium in an estimated 25% of hemangiomas, most hemangioendotheliomas, and most hemangiosarcomas. In conclusion, CD31 is a better marker than FVIIIra in the characterization of bovine endothelial tumors. The cell cycle regulatory pathways involving cyclin D1 and p53 seem to be impaired in endothelial urinary bladder tumors, p53 immunoreactivity positively correlating with enhanced invasion.

Immunohistochemical Evaluation of Vascular Urinary Bladder Tumors from Cows with Enzootic Hematuria

Immunohistochemical Evaluation of Vascular Urinary Bladder Tumors from Cows with Enzootic Hematuria

Bovine papillomavirus DNA in milk, blood, urine, semen, and spermatozoa of bovine papillomavirus-infected animals

Papillomavirus infection in bovines is associated with cutaneous papillomatosis on the hide, udders and other epithelial tissues, as well as in oral respiratory, alimentary and urinary tract mucosa. Bovine papillomavirus (BPV) is also considered the etiological agent of esophageal tumors and the malignant bladder tumors that characterize the clinical condition associated with chronic enzootic hematuria. After infective viral DNA was found in cattle blood and BPV1, 2 and 4 DNA in cattle reproductive and embryonic tissues, we looked for and found BPV DNA in blood, milk, urine, seminal fluid, and spermatozoa of BPV-infected animals. Peripheral blood lymphocyte cultures from BPV-infected animals had high rates of chromosome aberrations, including radial rearrangements that signal oncogenic potential and viral interaction with telomeric regions. The finding of BPV DNA in body fluids and tissues other than the epithelium demonstrates co-infection of other tissues or cell types by papillomavirus and shows the potential role of lymphocytes, seminal fluid and spermatozoa in BPV transmission. Our findings reinforce a peremptory need for prophylactic and therapeutic instruments to curtail this disease in bovine livestock.

Variation in ptaquiloside content in bracken (Pteridium esculentum (Forst. f) Cockayne) in New Zealand

AIM: To examine stands of bracken fern (Pteridium esculentum) from throughout New Zealand for the presence and concentration of ptaquiloside (Pta), and to compare the presence and/or concentrations of Pta in areas where bovine enzootic haematuria (BEH) and/or acute haemorrhagic syndrome (AHS) has been known to occur with those where BEH/AHS has not been recorded. METHODS: Stands of bracken fern were sampled from 275 sites throughout New Zealand. Sixty-two stands were from a regional survey predominantly from the Waikato and Coromandel regions, 27 were from a farm in the King Country where BEH/AHS had been investigated previously, and 186 were from a national survey of the North and South Islands. Sampling sites were from a mixture of grazed paddocks, roadsides, and forest and bush areas. Samples comprised whole young fronds, the tops of unfurling young fronds, or, for the Regional Survey, mature green fronds from the previous season. Pta was extracted from the samples, and measured using high-performance liquid chromatography. Information on the occurrence of BEH/AHS at specific locations was obtained from published information and records from animal health laboratories in New Zealand. RESULTS: The 275 samples contained widely varying concentrations of Pta. In the Farm Survey, concentrations ranged from 280–13,300 (mean 3,800) µg/g (on a dry-weight basis) in the 63% of samples that contained Pta. A high proportion of samples from the Regional and National Surveys covering large areas of the country contained no detectable levels of Pta. The majority (61%) of samples from these two surveys which contained Pta were from areas where BEH/AHS was reported to occur. Combining data from all surveys, in areas with reported BEH/AHS, 42% of samples collected contained Pta, compared with 6% where BEH/AHS was not known to occur. CONCLUSIONS: Concentrations of Pta in bracken in New Zealand vary greatly, and in a high proportion of stands Pta is not found. A higher incidence of Pta, and some very high concentrations, are found in areas where BEH/AHS was known to occur.

Detection of bovine papillomavirus type 2 in the peripheral blood of cattle with urinary bladder tumours: possible biological role

Bovine papillomavirus type 2 (BPV-2) infection has been associated with urinary bladder tumours in adult cattle grazing on bracken fern-infested land. In this study, we investigated the simultaneous presence of BPV-2 in whole blood and urinary bladder tumours of adult cattle in an attempt to better understand the biological role of circulating BPV-2. Peripheral blood samples were collected from 78 cattle clinically suffering from a severe chronic enzootic haematuria. Circulating BPV-2 DNA was detected in 61 of them and in two blood samples from healthy cows. Fifty of the affected animals were slaughtered at public slaughterhouses and neoplastic proliferations in the urinary bladder were detected in all of them. BPV-2 DNA was amplified and sequenced in 78 % of urinary bladder tumour samples and in 38.9 % of normal samples as a control. Circulating episomal BPV-2 DNA was detected in 78.2 % of the blood samples. Simultaneous presence of BPV-2 DNA in neoplastic bladder and blood samples was detected in 37 animals. Specific viral E5 mRNA and E5 oncoprotein were also detected in blood by RT-PCR and Western blot/immunocytochemistry, respectively. It is likely that BPV-2 can persist and be maintained in an active status in the bloodstream, in particular in the lymphocytes, as a reservoir of viral infection that, in the presence of co-carcinogens, may cause the development of urinary bladder tumours.

Epidemiological, clinical and paraclinical investigations in cattle enzootic haematuria in an area from N-W of Transilvania.

The observations were made on a number of 1751 cattle, during 2007-2008, coming from 7 localities, where the enzootic haematuria evolves, from county Maramures, next Baia Mare city (Sisesti, Negreia, Danesti, Surdesti, Plopis, Cetaţele, Budesti). Refering to feeding, in the winter period, natural hay and maize stalks and only exceptional maize and gran flour, and during grazing was low vegetation elected in the arid years predominantly from imperial fern (Pteridium aquilinum). The appearance of haematuria is in the january-february-march period, and appears frequently in pregnant animals and in animals around 2-3 years old. Individual and group clinical examinations from the examined localities praised poor state of maintenance, most common diseases associated with enzootic haematuria, and on the cows with clinically manifested haematuria, state of anaemia, haematuria, dysuria. We have collected 11 samples of blood, respectivelly 11 samples of urine by vesical catheterism from the cattle with clinical signs. Of them were made hemograma, complete leucograma and biochemical tests (serum glucose, BUN, total protein, albumin, gamaglobulin, creatinin). As far as urine: with color variations between yellow and straw yellow, orange and pale red, pH 2.0-8.0 and urinar sediment (vesical cells, red blood cells, calcium carbonate and sulfate crystals).