Enzootic Haematuria Research Articles

Prolonged detection of BPV2 in urine and blood of papillomatosis-affected cattle not grazing bracken fern.

BPV-2 infection can cause bladder infections in cattle that, when associated with bracken fern consumption, can progress to cancerous bladder tumors and also present as bovine enzootic hematuria (BEH). This study aimed to evaluate the prolonged natural BPV-2 infection in the blood and urine of cattle, excluding bracken fern consumption. Thirteen Girolando papillomatosis-affected cattle with no bracken fern contact history were monitored for 20 months. Blood, urine, and wart samples were collected for BPV-2 detection and clinical laboratory analyses. All animals showed the presence of BPV-2 in papillomas and blood, and 92.85% showed BPV-2 in urine, suggesting viral dissemination in the urinary tract. Despite all animals being infected with BPV-2, none showed BEH signs during the study. Thus, it was observed that BPV-2 infection alone didn't induce BEH over 20 months, implying a complex interaction with environmental factors or genetic predisposition. This underlines bracken fern consumption's critical role in urinary bladder carcinogenesis. The study underscores BEH's pathogenesis complexity, advocating longitudinal studies to comprehend BPV-2's role fully.

Ultrasonographic findings in the bladder of cattle at different stages of evolution of bovine enzootic hematuria in Brazil

Despite the diagnostic capacity of ultrasonography (US) in cases of bovine enzootic hematuria (BEH), it has been underused in the medical clinic for ruminants. Studies comparing ultrasound findings in healthy animals (HA) with subclinical (SCH) and clinical (CH) BEH are scarce in the literature. As a result, the present work aims to describe the US findings of cattle at different stages of BEH evolution, evaluating the diagnostic capacity and precocity of the technique. The study was carried out on five rural properties in the municipalities of Rio Preto and Bom Jardim de Minas – MG, Brazil, using 46 dairy cattle, females over 18 months of age, evaluated and classified as G1/CH, G2/SCH and G3/HA, according to history, clinical examination and urinalysis, with subsequent ultrasound analysis of the bladder. The mean bladder wall thickness was 36.50 mm in G1/CH, 33.53 mm in G2/SCH and 37.93 mm in G3/HA. Irregular walls were observed in 53.33 % (8/15) of G1/CH, 27 0.78 % (5/18) of G2/SCH and 38.46 % (5/13) of G3/HA. Loss of the architecture of the urothelium layers was observed in 53.33 % (8/15) of G1/CH, 33.33 % (6/18) of G2/SCH and 38.46 % (5/13) of G3/HA. Multifocal and Grade I lesion were the most common. Ultrasonography allowed the identification of lesions in 38.46 % of animals that did not present hematuria and in 33.33 % of animals that presented occult blood and proved to be an efficient technique for detecting pre-clinical cases of BEH, allowing easy identification. and early onset of BEH cases.

Concentrações de haptoglobina em vacas com histórico de ingestão crônica de samambaia

ABSTRACT: Bracken ferns (Pteridium spp.) have been associated with intoxication in cattle and is the cause of bovine enzootic hematuria (BEH) characterized by intermittent hematuria, anemia, and poor body condition. Acute phase proteins have not previously been investigated in detail in BEH. Thus, this investigation determined serum concentrations of haptoglobin, selected hematological (complete blood count and fibrinogen), biochemical (total serum protein, albumin, urea, creatinine, aspartate aminotransferase, gamma glutamyl transferase, and creatine kinase), and urinary analytes, in 50 Nelore cows from farms with history of natural bracken fern chronic ingestion in the southern region of Brazil. Twenty-nine cows had macrohematuria and 21 microhematuria. From the 29 cows with macrohematuria, 13 had a countless number of erythrocytes per high power macroscopic field and anemia. All cows had dipstick proteinuria. Median haptoglobin concentrations were elevated for 47 BEH cows with a 3.9-fold increase, suggesting a chronic inflammatory response while the number of total leukocytes and segmented neutrophils were above the reference intervals for 14 cows. Besides inflammation, there is a hypothesis that bracken fern could cause hemolysis in the circulation. Therefore, there could be a larger acute phase reaction but much of the haptoglobin was being removed as it was being produced. In our study, increases in haptoglobin, creatine kinase, hyperglobulinemia and anemia were found in cows with BEH, contributing to the understanding of the chronic inflammatory response which could be associated with neoplastic lesions in the urinary bladder caused by long-term and repeated ingestion of Pteridium species.

Bracken poisoning

Bracken (Pteridium species) is an ancient, large and very widespread fern that has long been recognised as poisonous. It contains several toxic compounds including ptaquiloside and thiaminase. Bracken toxicosis manifests as different clinical syndromes in different animals. Cattle develop an acute haemorrhagic syndrome leading to widespread haemorrhage and death. With more chronic ingestion enzootic bovine haematuria occurs, which is characterised by tumours of the upper alimentary tract and urinary bladder. Sheep also develop acute haemorrhage syndrome and cancer, but chronic bracken ingestion causes retinal neuropathy, which can lead to permanent blindness. Bracken toxicosis in monogastric animals such as pigs is because of an induced avitaminosis B1. Diagnosis of bracken toxicosis is based on clinical signs, history of exposure and characteristic findings at postmortem. In cattle and sheep the clinical syndromes are progressive and humane euthanasia is usually necessary in animals with advanced disease. In pigs with bracken toxicosis signs are often non-specific and lead to sudden death, but supplementation with thiamine can be effective.

Hematúria enzoótica bovina no estado do Tocantins e o uso da ultrassonografia como método de diagnóstico

Bovine enzootic hematuria (BEH) is a chronic disease caused by ingestion of Pteridium esculentum subsp. arachnoideum (Kaulf.) Thomson. This disease has great importance because it affects the animal wellbeing, causes economic losses, and hinders public health due to the toxic principle that is present in the milk of cows that ingest this plant. Early diagnosis assists in the decision making for disposal and replacement of animals in endemic regions. Despite ultrasonography is a useful tool for diagnosis of this disease, there is a lack of studies evaluating BEH in the state of Tocantins, Brazil, using ultrasound and epidemiological data and clinical signs; however, the histopathological examination is the method that provides the definitive diagnosis. The objective of this study was to describe the occurrence of BEH in the state of Tocantins and evaluate the efficiency of bladder ultrasound examination for diagnosis and prognosis of BEH. Ten animals that presented intermittent hematuria and hypo-colored mucous membranes were subjected to clinical and complementary exams. The urine physical analysis presented light red color, high turbidity, and positive sedimentation. The ultrasonography showed a thick, irregular mass in the bladder wall, containing an echogenic liquid of high cellularity and pedunculated papillary lesions. Macro and microscopic analyses of the animals’ bladders showed areas with vascular proliferation in the sub-mucous membranes associated with mononuclear inflammatory cell infiltrate and proliferation of the epithelium. Ultrasonography is an excellent option and, combined with clinical and laboratory exams of blood and urine, enables the diagnosis and prognosis of BEH.

Ocurrence enzootic hematuria in buffaloes in Brazil: epidemiological, clinical, and pathological aspects

ABSTRACT: It is reported the occurrence of enzootic hematuria (EH) in buffaloes in Brazil after performing an epidemiological survey and clinicopathological analises. To date, EH caused by ingestion of Pteridium esculentum subsp. arachnoideum, a radiomimetic plant popularly known as “bracken fern”, has not been described in this species in Brazil. Bovine EH is responsible for high economic losses in Brazil’s Southeast Region not only because of the deaths it causes, but also owing to its negative effect on productivity. In São José do Barreiro County, São Paulo, some farmers in areas with a high incidence of bovine EH have been replacing cattle with buffaloes, based on the premise that the latter would be more resistant to poisoning by ingestion of Pteridium spp. However, even though initial observations indicated that buffaloes are indeed less sensitive than cattle to the toxic principle of Pteridium spp., cases of hematuria in this species have been reported. According to preliminary date, EH only occurs in buffaloes over six years of age. Macroscopic examination revealed a thickened urinary vesicle mucosa, along with multiple foci of ulcerated, exophytic, verrucous, and pedunculated lesions. In one of the buffaloes studied, the bladder wall was ruptured and exhibited marked secondary inflammation. Histologically, neoplastic and non-neoplastic changes similar to those described in cattle poisoned by Pteridium spp. were observed. The neoplasms found included papilloma, carcinoma in situ, urothelial carcinoma (low and high grade), inverted, microcystic, and trabecular variants, urothelial carcinoma with divergent differentiation (squamous and glandular), squamous cell carcinoma, lymphangioma, hemangioma, and hemangiosarcoma. There was also coexistence of epithelial and mesenchymal neoplasms. Bovine papillomavirus particles were not detected by polymerase chain reaction in the bladder samples analyzed.

Botanical characteristics, toxicity and control of bracken fern (pteridium aquilinum (l.) kuhn)

Bracken (Pteridium aquilinum (L.) Kuhn) is cosmopolitan fern, found at varying altitudes on all continents except Antarctica. It is an aggressive colonizer and can appear in various plant communities, but an important characteristic is its ability to dominate in dense patches. Once establish, the deep-set rhizomes are nearly impossible to eradicate. In Croatia, bracken dominate on specific habitat type called “bujadnice” in Lika region, but can also be found in herbaceous layers in following forest communities: As. Betulo-Quercetum; As. Pteridio-Betuletum; As. Potentillo albae-Quercetum pubescentis; As. Castaneo sativae-Fagetum; As. Helleboro nigri-Piceetum. Bracken fern is significant problem for livestock-based extensive agriculture, because it causes a range of syndromes in farm animals including thiamine deficiency, acute hemorrhagic syndrome, bright blindness, enzootic hematuria and upper alimentary carcinoma. Man may consume the toxins of bracken either directly or indirectly. In terms of the economic loss and suffering caused by bracken, it is clearly that some steps must be taken to control distribution of this fern. Timing is important in any management treatment of bracken fern. The most effective time for bracken control is summer just after the new fronds have fully expanded and starch reserves in the rhizome are at their lowest level. Two or more annual treatments and combinations of cutting and herbicide are more effective than single treatments or even single annual treatments.

Detection and Quantification of the E6 Oncogene in Bovine Papillomavirus Types 2 and 13 From Urinary Bladder Lesions of Cattle.

Bovine papillomavirus types 2 and 13 can induce tumors in both the cutaneous and mucosal epithelia of cattle. These viral types are associated with the development of benign cutaneous papillomas and malignant lesions in the urinary bladders of cattle, with the latter being known as bovine enzootic hematuria. Among the viral oncoproteins encoded by Deltapapillomavirus DNA, the E6 oncoprotein has an important role in cell proliferation and might be related to cancer initiation and promotion. The aim of this study was to present a standardized SYBR Green-based quantitative PCR for detection and quantification of the bovine papillomavirus 2 and 13 E6 oncogenes in urinary bladder samples from cattle. Twenty-four urinary bladders from cattle displaying tumors (n = 12) and normal bladder mucosa (n = 12) were tested by quantitative PCR. Of the 12 urinary bladders with tumors, six presented bovine papillomavirus 2 DNA concentrations ranging from 1.05 × 104 to 9.53 × 103 copies/μL, while two had bovine papillomavirus 13 DNA amplified at concentrations of 1.30 × 104 to 1.23 × 104 copies/μL. The healthy bladder mucosa samples were negative for both bovine papillomaviruses. Once the results were confirmed by conventional PCR and direct sequencing, the quantitative PCR assay developed in this study was shown to be a sensitive and specific tool for detecting and quantifying the E6 ORF of bovine papillomavirus 2 and 13 in a variety of clinical samples. Our findings of identification of bovine papillomavirus 2 and 13 DNA in urothelial tumors from cattle suffering from bovine enzootic hematuria agree with data from previous studies, representing the first detection of bovine papillomavirus 13 DNA in malignant bladder lesions of cattle from Brazil.

Digital droplet PCR for the detection and quantification of circulating bovine Deltapapillomavirus.

In this study, the digital droplet polymerase chain reaction (ddPCR) was used to quantify circulating bovine papillomavirus (BPV; genus: Deltapapillomavirus) DNA levels in blood samples from 25 clinically normal cows and 15 cows with chronic enzootic haematuria due to papillomavirus-associated bladder tumours. ddPCR detected BPV DNA in 95% of all the samples (i.e. in 24 of the clinically normal cows and 14 of the diseased animals), whereas quantitative real-time PCR (qPCR) detected it in only 57.5% of the same blood samples, with percentage differences between ddPCR and qPCR being statistically significant (p-value≤.05), according to chi-squared test. Furthermore, ddPCR detected BPV infections by a single genotype and by multiple genotypes in 37% and 63% of the cows, whereas qPCR detected these in 16% and 16%. Of the two assays, ddPCR was the more sensitive and accurate clinical diagnostic tool, allowing the detection of otherwise undetectable BPV genotypes, and consequently, a higher number of BPV co-infections. qPCR failed to detect many BPV co-infections by multiple genotypes. Therefore, ddPCR may be an essential tool for improving diagnostic procedures, allowing the identification of the genotypic distribution of BPV and a better understanding about the territorial divergence, if any, of the BPV prevalence in different areas. No significant differences in the blood viral load estimations were observed between the two animal groups, suggesting that the bloodstream could be a site of primary infection. Finally, as BPV DNA was detected in cows affected by non-invasive urothelial tumours, including papilloma and papillary urothelial neoplasms of low malignant potential, the circulating BPVs appeared to be independent of the status of urothelial neoplasms. Therefore, unlike in humans, circulating BPVs cannot be an actual prognostic marker of urothelial tumours in cows.

Ptaquiloside and Pterosin B Levels in Mature Green Fronds and Sprouts of Pteridium arachnoideum.

Pteridium arachnoideum, a fern of the Pteridium aquilinum species complex found in South America, is responsible for several different syndromes of poisoning. Cases of bovine enzootic hematuria and upper alimentary squamous cell carcinoma are both frequent occurrences in Brazil, whereas only bovine enzootic hematuria is noted with any frequency around the world. The reason for the high frequency of upper alimentary squamous cell carcinoma in Brazil is not currently known. One possible explanation may be the higher levels of ptaquiloside and pterosin B in Brazilian Pteridium than those present in the plant in other countries. However, these levels have not yet been determined in P. arachnoideum. Thus, the present study aimed to measure and compare ptaquiloside and pterosin B levels in mature green fronds and sprouts of P. arachnoideum collected from different locations in Brazil. Samples of P. arachnoideum were collected from the states of Minas Gerais and Rio Grande do Sul. A total of 28 mature leaf samples and 23 sprout samples were used. The mean concentrations of ptaquiloside and pterosin B present in the mature green fronds of P. arachnoideum ranged from 2.49 to 2.75 mg/g and 0.68 to 0.88 mg/g, respectively; in P. arachnoideum sprouts, mean concentrations of ptaquiloside and pterosin B ranged from 12.47 to 18.81 mg/g, and 4.03 to 10.42 mg/g for ptaquiloside and pterosin B, respectively. Thus, ptaquiloside and pterosin B levels in P. arachnoideum samples collected in Brazil were higher in sprouts than in mature green fronds, as observed in other countries. However, there was no variation in ptaquiloside levels among plants collected from different cities in Brazil. The high frequency of upper alimentary squamous cell carcinoma in Brazilian cattle may not be attributed to greater levels of ptaquiloside and pterosin B in P. arachnoideum than in other Pteridium species in other countries.

Morphometric and Vascular Analyses and MMP-2 Expression in Bladders of Animals with Bovine Enzootic Haematuria

Background: Bovine enzootic haematuria is a chronic, non-infectious disease caused by the ingestion of bracken Pteridium aquilinum that contain ptaquiloside, substance responsible for the development of neoplasms in combination with bovine papillomavirus infection. Studies examining metalloproteinases have shown that these enzymes may be useful as prognostic markers and many studies have correlated the intratumoural microvessel density and incidence of metastasis in a variety of cancers, including bladder cancer. This study aimed to evaluate the urothelial morphology and vasculature and quantify the expression of the enzyme metalloproteinase-2 in the bladder lesions of bovine with enzootic haematuria.Materials, Methods & Results: Forty bladders with macroscopic lesions were subjected to routine histological processing. The material was stained using the Hematoxylin-Eosin (HE) and Weigert's resorcin-fuchsin staining methods. The morphometric analysis included the measurement of the total thickness of the urothelium. Microscopically, the lesions were differentiated as neoplastic or non-neoplastic, and the vascular changes were characterised as vascular proliferation, ectasia, dilation and thickening and the material stained using the Weigert's resorcin-fuchsin method was used to evaluate sclerosis and the vascular fibroelastosis. The metalloproteinase expression was evaluated using an anti-metalloproteinase-2 antibody. The main non-neoplastic lesions found in the urothelium included clear cell metaplasia, dysplasia, hyperplasia, haemorrhage, cystitis cystica and Brunn's nests and the neoplastic lesions were haemangioma, myxoma, transitional cell carcinoma, adenocarcinoma, in situ carcinoma and haemangiosarcoma. The urothelium measurements revealed differences between the epithelial thicknesses of bladders with and without neoplasms. The bladder vasculature evaluation revealed the frequent occurrence of different pathological aspects, such as vascular proliferation, dilation, expansion, ectasia and thickening; more severe vascular proliferation, thickening and fibroelastosis were observed in bladders with neoplasms. Metalloproteinase-2 expression was observed in the epithelial cells, fibroblasts, endothelial cells and smooth muscle cells present in the tumour tissue and was a significant increase in the expression of MMP-2 in the neoplasms of mesenchymal origin compared to the neoplasms of epithelial origin.Discussion: The presence of a large variety of non-neoplastic lesions indicates the possibility that there are other factors concomitant to BEH involved. Epithelial hyperplasia and metaplasia, for example, could be the result of damage caused by bovine papillomavirus type 2 or even an altered urine pH. Furthermore, it is believed that the bladder has the capacity to generate various types of neoplasms due to the different histological components of the bladder, thus enabling it to generate both mesenchymal and epithelial lesions. The thickness of the urothelium was greater in all the quadrants with neoplasms compared to the quadrants without neoplasms. These data revealed that it is possible to use less invasive diagnostic imaging methods to characterise bovine enzootic haematuria and to measure tumour progression based on the severity of vascular changes. The vascular proliferation and thickening were significant in bladders with neoplasms, which may be explained by the fact that tumours require an extensive vascular supply. In addition, many tumours release vascular growth factors and promote neovascularisation. The intensity of the immunostaining of metalloproteinase-2 differed among the different types of neoplasms. However, due to the diversity of the tumours, it was not possible to establish a relationship between metalloproteinase-2 expression and tumour progression.

Diagnosis of the incidence of enzootic Hematuria Bovina of bovinos in production of three winning areas

Enzootic bovine hematuria (HEB) is a disease caused by exposure to, and consumption of pastures invaded by common fern (Pteridium aquilinum), also known as llashipa. The objective of the research was to identify the cattle areas of Cantón Chilla, establishing a relationship between their location and the incidence of enzootic bovine hematuria. Three zones were studied, each with four sectors, which were grouped according to their height - Zone A (300 - 999 masl), Zone B (1000 - 1999 masl) and Zone C (2000 - 3300 masl). The presence of llashipa in the surveyed areas corresponds to: 20% in Zone A, 60% in Zone B, and 50% in Zone C; data corresponding to recurrent invasive weeds in pastures. The urinary test strip techniques (TRU) and microscopic analysis of the urinary sediment (AMSU) were applied for the analysis of the samples. The results found were 36.1% (130/360) for AMSU and 5.28% (19/360) for TRU; whereupon the two methods demonstrate the existence of this disease in the canton of Chilla. Regarding the relation of the cellular elements variables (AMSU and TRU) and livestock areas (A, B and C), by means of the Chi square statistic test, it shows that the significance value p is less than 0.005, which indicates that there is a connection between the variables. It is concluded that the sectors grouped in the different livestock areas of Canton Chilla are endemic to enzootic bovine hematuria according to the height at which they are located.

Determination of ptaquiloside in cow's milk by HPLC-UV

Ferns of the genus Pteridium are known to be responsible for cases of poisoning in cattle. The commonly found species of Pteridium in Brazil are P. arachnoideum and P. caudatum. The toxic principle of these ferns is ptaquiloside, a norsesquiterpene glucoside, which is a known carcinogen. Ptaquiloside can be secreted in the milk of cows grazing on bracken ferns, thereby posing a potential health risk for the milk consumers. The objective of the present study was to develop an analytical methodology using high performance liquid chromatography (HPLC) with ultraviolet (UV) detection to determine the concentrations of ptaquiloside in cow´s milk. HPLC-UV method, based on the conversion of ptaquiloside into pterosin B and its subsequent extraction using dichloromethane, was successfully developed for estimating concentrations of ptaquiloside in milk samples. However, the presence of ptaquiloside was not detected in milk samples obtained from cows with a history of bovine enzootic hematuria resulting from grazing on P. arachnoideum infested pastures in Brazil.

Allyl-isothiocyanate ameliorates the pre-neoplastic changes induced by the fern Dryopteris nigropalaceae on experimental feeding in Guinea pigs

Enzootic bovine haematuria, caused by long-term ingestion of ferns, is a chronic disease of hill cattle characterized by neoplastic lesions in the urinary bladder. Objectives of this study were to investigate the toxicity potential of long-term feeding of the fern Dryopteris nigropalaceae and effect of allyl isothiocyanate (AITC) to ameliorate fern toxicity and the associated pathological changes. The LC-MS analysis of the fern showed presence of ptaquiloside (4.5 ± 1.0 μg/g) and pterosin B (39 ± 9.1 μg/g). Groups of animals were fed dried fern powder at the dose of 20% w/w in normal feed and treated with and without AITC at graded doses. Long term feeding of fern induced inflammatory and pre-neoplastic lesions in urinary bladder. The important lesions included cystitis, squamous metaplasia and high-grade dysplasia. Urothelium showed positive immunoreactions for nuclear expression of H-ras and p53. However, no mutation suggestive of neoplastic change was observed on partial mRNA sequences analyses of exon 2 of H-ras and 5 or 7&8 of p53 genes. Strikingly, AITC showed dose-dependent amelioration of pre-neoplastic changes in fern-fed animals. In conclusion, AITC is shown to limit pre-neoplastic changes caused by D. nigropalaceae feeding in guinea pigs.

Bovine papilloma: aetiology, pathology, immunology, disease status, diagnosis, control, prevention and treatment: a review

Bovine papillomavirus (BPV) induces diseases of economic and veterinary importance leading to significant economic losses to livestock owners such as leather depreciation and mortality when it progresses to neoplasms. BPV affects the spinosum and granulosum layers of the skin causing warts. There are many reportage of BPV infection in domestic animals such as cattle, birds and wild animals such as tapir, giraffe, antelope and zebras. It also causes equine sarcoids in horses and donkeys. Gross lesions seen are mainly benign papillomas which most times regress but occasionally persist and progress to malignancy of the urinary bladder and upper alimentary tract leading to Enzootic haematuria. Equine sarcoids could be of the occult, nodular or verrucous types. Due to the ability to BPV to cause cancers, haematuria and skin pathologies, diagnosis of this infection has attracted huge interests. Diagnostic techniques include clinical examination, histopathology, polymerase chain reaction (PCR) Southern blot, dot blot, reverse blot, in situ hybridization and immunohistochemistry. All the aforementioned have been very helpful in the diagnosis of BPV. In conclusion, this review focused on the aetiology, viral structure pathogenicity, transmission, macroscopic and microscopic pathology, pathogenesis, immunology, gene products, prevention, treatment and control of this virus in affected animal species.

The role of inducible nitric oxide synthase in urinary bladders of cattle with enzootic hematuria and its struggle with uroplakin III

Changes in uroplakin secretion in the urothelium leads a gradual ability lost in neoplastic conditions. Urothelium is one of the target regions where nitric oxide can damage. Therefore, in this study, the presence of iNOS and UPIII were investigated in bovine enzootic hematuria cases. The results were supported by comparing them with those achieved by critical markers for malignancy of tumor such as caspase 3, Bax, p53, PCNA, and VEGF. The presence of 8-OHDG, a good marker for oxidative DNA damage, was also investigated. For this purpose, urinary bladders with enzootic hematuria were collected from the slaughterhouses. In histopathological examinations, neoplastic, pre-neoplastic, and inflammatory lesions were observed in these urinary bladders. In immunohistochemical examinations, iNOS reacts as negative in the UPIII positive healthy urothelium which preserves the structure of AUM. On the other hand positive reaction has been observed in AUM modified and UPIII negative urothelium. In these cases, as an indicator of oxidative damage, 8-OHdG positivity was observed in the UPIII negative and iNOS positive areas. Additionally, increased cell proliferation, decreased apoptosis as well as p53 disorganization were clearly observed in most of the cases. Thus, the active role of nitric oxide, as a free radical derivative, has been shown in the enzootic hematuria cases of cattle.

Diagnóstico Clínico de la Hematuria Vesical Enzoótica Bovina por Urianálisis de la Provincia de Oxapampa, Perú

El estudio se desarrolló en la provincia de Oxapampa, Región Pasco, Perú, con el objetivo de diagnosticar clínicamente la hematuria vesical enzoótica bovina (HVEB) mediante el urianálisis, acompañado de la evaluación clínica y semiológica del animal y de la inspección general del establecimiento ganadero. Se obtuvieron muestras de orina por estimulación manual externa perivulvar y perineal de 211 bovinos hembras, mayores de 6 meses, expuestos naturalmente al consumo del helecho común o Pteridium aquilinum (Pa). Las muestras recolectadas fueron sometidas a una prueba tamiz con tiras reactivas (TR), evaluación físico-química (color, turbidez, peso específico y pH), pruebas complementarias químicas (proteína, glucosa y bilirrubina), así como una evaluación microscópica del sedimento (EMS), prueba confirmatoria de hematuria, relevante para los casos subclínicos (microhematuria). Mediante la EMS, los casos de hematuria presentaron variadas intensidades, por lo cual se planteó una clasificación de grados de acuerdo a ellas, basado en el número de eritrocitos por campo hallados en cada muestra. Se discute la relación entre los resultados de la EMS con las TR, pruebas físico-químicas y químicas complementarias. Se encontró una prevalencia de 26.1% para la zona y los resultados del análisis estadístico indican que la edad y el intervalo al último parto pueden ser considerados como factores predisponentes de la HVEB.

Survey of ferns and clinico-pathological studies on the field cases of Enzootic bovine haematuria in Himachal Pradesh, a north-western Himalayan state of India

Enzootic bovine haematuria (EBH) in cattle occurs in upland areas of the world. In India, the disease is present in isolated pockets in the Himalayas and in the Nilgiri Hills. The variation in the disease incidence has been attributed to different environmental conditions and animal rearing practices followed in the different regions. The aim of the study was to conduct field surveys in parts of EBH endemic regions of Himachal Pradesh, a north-western Himalayan state of India. Out of the total 103 plant samples collected, a total of 95 samples were identified as ferns. The major ferns identified included, Onychium japonicum (Thunb.) Kunze, Polystichum piceopaleaceum Tagawa, Dryopteris juxtaposita Christ, Pseudocyclosorus canus (Baker) Holttum and J.W. Grimes, Onychium contiguum C. Hope, Dryopteris nigropaleacea (Fraser-Jenk.), Pteridium aquilinum (L.) Kuhn, Diplazium esculentum (Retz.) Sw., Allantodia maxima (D. Don) Ching, Woodwardia unigemmata (Makino) Nakai, Pteris cretica L., Pteris vittata L., Asplenium trichomanes L., Thelypteris phegopteris (L.) Sloss. ex Rydb, Adiantum venustum D. Don and Paraceterach vestita (Hook.) R.M. Tryon. The concentration of ptaquiloside (PTA) and pterosin B (PtB) in some of the ferns collected from Kullu and Chamba regions ranged from 0 to 358.6 ± 70.5 μg/g and 0 to 652.4 ± 50.0 μg/g, respectively. In addition, field cases of the disease in cattle were also studied in the EBH endemic districts. A total of sixteen cattle urine samples and one urinary bladder of EBH affected cattle were collected. On physical, chemical (benzidine test) and microscopic examination of urine sediment, all the sixteen field samples were found to be positive for erythrocytes and the cases were diagnosed as macrohaematuria. The clinico-pathological studies on the field cases and the presence of PTA and PtB in the ferns indicated that EBH is a prevalent disease and there is an association between chronic fern ingestion and EBH in cattle. On the basis of gross pathology, histopathology and immunohistochemistry (p53 and H-ras nuclear expression in the urothelial cells) of the urinary bladder tissue, the field case was diagnosed as transitional cell adenocarcinoma with chronic lymphocytic cystitis.

Screening for Ptaquiloside in Ferns: Using Herbarium Specimens for Qualitative Mapping Purposes

Ptaquiloside (PTA) is a noxious carcinogen found widespread in Bracken (Pteridium sp.) but with scattered and unresolved distribution outside the genus. The carcinogen causes Bovine Enzootic Haematuria among cattle all-over the World and is under suspicion of causing human cancers. To set-up a methodology for large-scale qualitative studies on the distribution of PTA in ferns using already available herbarium specimens as source. PTA and the main degradation product pterosin B (PtB) were quantified in aqueous frond extracts by HPLC-DAD. PTA was quantified after forced reaction into PtB. Optimal reaction conditions were tested using hydrochloric acid, sulphuric acid, trifluoroacetic acid (TFA), acetic acid and temperature as the experimental variables. A pair-wise test for PTA degradation in plant-press were used to explore the effect of this preservation regime. A selection of up to 50years old Bracken herbarium specimens were tested for PTA and PtB. The methodology was applied on 21 fern species from Denmark. An optimised TFA-based method results in 30.7% higher conversion and a 1:1 reaction between PTA and PtB. Full three-dimensional resolution of the analyte was obtained. Preservation of fronds in a plant press increase formation of PtB. Hence, the method is only suitable for qualitative studies. Presence of PTA and PtB were found in samples up to 50years old. Among 21 ferns tested, the compounds were only found in Pteridium aquilinum. Herbarium specimens up to 50years old can be used for explorative risk assessment of ferns using HPLC-DAD for quantification and identification. Copyright © 2017 John Wiley & Sons, Ltd.

Bovine enzootic haematuria from consumption of Pteris deflexa and Pteris plumula in northwestern Argentina

Bovine enzootic haematuria (BEH) is caused by prolonged ingestion of toxic principles of bracken fern, essentially by Pteridium spp. In northwestern Argentina, this disease has a great economic impact ant it is attributed a chronic consumption to Pteridium arachnoideum. This paper describes two endemic areas for enzootic hematuria due to the consumption of Pteris deflexa and Pteris plumula. Two areas where P. deflexa and P. plumula are endemic, but free of Pteridium species, were devised and seven farms were visited. The disease was confirmed based on the presence of clinically affected animals. In four necropsies bleeding neoplastic lesions were observed in the mucosa of the urinary bladder. At phytochemical analysis, both ptaquiloside and pterosin B were found in P. deflexa and P. plumula. Thus, the consumption of P. deflexa and P. plumula can also cause BEH.