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.

The role of bracken fern illudanes in bracken fern-induced toxicities.

Bracken fern is carcinogenic when fed to domestic and laboratory animals inducing bladder and ileal tumours and is currently classified as a possible human carcinogen by IARC. The carcinogenic illudane, ptaquiloside (PTQ) was isolated from bracken fern and is widely assumed to be the major bracken carcinogen. However, several other structurally similar illudanes are found in bracken fern, in some cases at higher levels than PTQ and so may contribute to the overall toxicity and carcinogenicity of bracken fern. In this review, we critically evaluate the role of illudanes in bracken fern induced toxicity and carcinogenicity, the mechanistic basis of these effects including the role of DNA damage, and the potential for human exposure in order to highlight deficiencies in the current literature. Critical gaps remain in our understanding of bracken fern induced carcinogenesis, a better understanding of these processes is essential to establish whether bracken fern is also a human carcinogen.

Intoxicação crônica por Pteridium arachnoideum em bovinos no nordeste do estado do Rio Grande do Sul, Brasil / Chronic poisoning by bracken fern (Pteridium arachnoideum) in cattle in the northeast region of the state of Rio Grande do Sul, Brazil

Pteridium spp., popularmente conhecida como samambaia, comumente encontrada em solos ácidos, arenosos e de baixa fertilidade. A intoxicação ocorre quando os animais passam por privação alimentar ou ingestão acidental. O objetivo deste estudo é relatar a ocorrência de dois casos de intoxicação crônica por samambaia em bovinos na região nordeste do estado do Rio Grande do Sul, Brasil. Ambos os casos cursaram com sinais clínicos de disfagia, halitose, salivação e presença de tumores na orofaringe. A baixa oferta e qualidade de forragem, superlotação das áreas de pasto, manejo inadequado das pastagens, a presença de Pteridium arachnoideum, os sinais clínicos em conjunto com os achados de necropsia e o exame histopatológico confirmaram o diagnóstico de intoxicação crônica.

Pteridium spp., commonly known as fern, is commonly found in acidic, sandy and low fertility soils. Poisoning occurs when animals go through food deprivation or accidental ingestion. The objective of this study is to report the occurrence of two cases of chronic intoxication in cattle in the northeast region of the state of Rio Grande do Sul, Brazil. Both cases had clinical signs of dysphagia, halitosis, salivation and the presence of tumors in the oropharynx. Low supply and quality of forage, overcrowding of pasture areas, inadequate pasture management, presence of Pteridium arachnoideum, clinical signs, necropsy and histopathological of lesions confirmed the diagnosis of chronic intoxication.

Intoxicação crônica por Pteridium arachnoideum em bovinos no nordeste do estado do Rio Grande do Sul, Brasil / Chronic poisoning by bracken fern (Pteridium arachnoideum) in cattle in the northeast region of the state of Rio Grande do Sul, Brazil

Pteridium spp., popularmente conhecida como samambaia, comumente encontrada em solos ácidos, arenosos e de baixa fertilidade. A intoxicação ocorre quando os animais passam por privação alimentar ou ingestão acidental. O objetivo deste estudo é relatar a ocorrência de dois casos de intoxicação crônica por samambaia em bovinos na região nordeste do estado do Rio Grande do Sul, Brasil. Ambos os casos cursaram com sinais clínicos de disfagia, halitose, salivação e presença de tumores na orofaringe. A baixa oferta e qualidade de forragem, superlotação das áreas de pasto, manejo inadequado das pastagens, a presença de Pteridium arachnoideum, os sinais clínicos em conjunto com os achados de necropsia e o exame histopatológico confirmaram o diagnóstico de intoxicação crônica.

Pteridium spp., commonly known as fern, is commonly found in acidic, sandy and low fertility soils. Poisoning occurs when animals go through food deprivation or accidental ingestion. The objective of this study is to report the occurrence of two cases of chronic intoxication in cattle in the northeast region of the state of Rio Grande do Sul, Brazil. Both cases had clinical signs of dysphagia, halitosis, salivation and the presence of tumors in the oropharynx. Low supply and quality of forage, overcrowding of pasture areas, inadequate pasture management, presence of Pteridium arachnoideum, clinical signs, necropsy and histopathological of lesions confirmed the diagnosis of chronic intoxication.

Achados clínico-patológicos e métodos de controle da intoxicação por Pteridium (aquilinum) arachnoideum em uma propriedade do Rio Grande do Sul / Clinical and pathological findings and control methods of the poisoning by Pteridium (aquilinum) arachnoideum in a farm of Rio Grande do Sul, Brazil

A infestação de pastagens por Pteridium arachnoideum é um problema que afeta a pecuária em diversas partes do mundo, ocasionando perdas produtivas e mortalidades nos rebanhos, bem como a redução das áreas de pastagens. Devido aos prejuízos na bovinocultura, foi proposto o acompanhamento de uma propriedade no Rio Grande do Sul com perdas por Pteridium arachnoideum. Realizou-se um levantamento das mortes associadas à intoxicação por samambaia na propriedade durante o período de janeiro de 2007 a janeiro de 2015. Adicionalmente, testou-se a viabilidade de ovinos como ferramenta de controle biológico de samambaia e se acompanhou as tentativas de controle da planta através do uso dos herbicidas metilsulfuron-metil e picloram, os quais eram associados ou não à roçada prévia. Das mortes por intoxicação natural acompanhadas na propriedade, observou-se 22 casos de quadro agudo de diátese hemorrágica e seis de quadro crônico de carcinomas do trato digestório superior. Em cinco bovinos jovens com diátese hemorrágica, além de hemorragias disseminadas e infartos, observou-se acentuado edema laríngeo, que cursava, clinicamente, com dispneia e estertores respiratórios característicos. Os carcinomas do trato digestório superior, apesar de menos frequentes, causaram perdas expressivas, devido à mortalidade anual de reprodutoras. Notou-se que a introdução de bovinos jovens em áreas recentemente roçadas pode resultar no consumo de grandes quantidades de samambaia e na ocorrência de surtos da enfermidade aguda. O controle das populações de samambaia pelo pastejo por ovinos não foi eficiente, devido ao baixo consumo observado com a lotação de quatro ovinos por hectare, embora mortalidade de ovinos por consumo da planta não tenha sido registrada. Após a retirada do potreiro infestado por samambaia, ao final do experimento, oito dos ovinos foram acompanhados, clinicamente, por três anos e não apresentaram nenhuma alteração. A utilização dos herbicidas resultou em uma redução da cobertura de P. arachnoideum. No entanto, essa prática necessita de estudos adicionais, já que algumas áreas não tiveram recuperação satisfatória da pastagem e o seu uso pode causar impactos ambientais e aquisição de resistência de P. arachnoideum aos herbicidas.(AU)

The infestation of pastures by populations of Pteridium arachnoideum has been a considerable and global problem to the livestock production, due to animal mortality and pasture coverage reduction. Given the impact of P. arachnoideum on cattle production, it has been proposed to monitor a beef cattle farm in Rio Grande do Sul, to assess some of the losses associated with the plant consumption and the methods employed for the plant control. A survey of cattle deaths associated with fern poisoning on the farm was carried out from January 2007 to January 2015. In addition, the viability of sheep as a biological control tool was tested. Attempts to control the plant through the use of the herbicides methylsulfuron-methyl and picloram associated or not with previous mowing were monitored. Cases of natural poisoning observed in the farm included the acute form (22), known as hemorrhagic diathesis as well as the chronic form, consisting in digestive carcinomas (6). Five cases of hemorrhagic diathesis in young cattle went along with marked laryngeal edema, which was clinically manifested as dyspnea and roaring, in addition to the classic pathological changes of widespread hemorrhages and infarcts. Even though less frequent, upper digestive tract carcinomas caused significant losses, due to annual mortality of mature cows. In occasions, when cattle were moved to newly mowed infested areas, outbreaks of acute poisoning were observed. The attempted control by sheep introduction showed to be unpractical, due to the little consumption seen at a stocking rate of four sheep per hectare. However, sheep mortality due to P. arachnoideum consumption was not recorded. The plant control method applied at the farm is based on both, the mowing of densely populated areas and the use of herbicides, namely metylsulfuron-methyl and picloram, in all paddocks. The reduction of P. arachnoideum coverage showed to be varied in different paddocks, however allowed the recovery of some grazing areas. This practice needs further studies, as some areas have not had satisfactory pasture recovery, and its indiscriminate use could lead to environmental impacts and resistance of P. arachnoideum to herbicides.(AU)

Timpanismo ruminal crônico secundário à hipoplasia de pregas de omaso em dois bovinos / Chronic ruminal bloat secondary to omasal laminae hypoplasia in two cattle

Um novilho normando e outro charolês apresentando distensão abdominal, diarreia intermitente e timpanismo ruminal crônico, que iniciaram após desmame, foram enviados para necropsia. Observou-se ausência de pregas omasais associada à hipoplasia do órgão, assim como redução de tamanho das papilas ruminais e reticulares. Com base nas lesões e histórico, conclui-se que o timpanismo ruminal foi ocasionado pela falha no desenvolvimento do omaso...

Two emaciated juvenile steers, one Normande and one Charolaise breed with abdominal distension, intermittent diarrhea and chronic ruminal bloat that had begun at weaning were necropsied. Absence of the omasal laminae with omasal hypoplasia were found together with loss of ruminal papillae and reticular folds. Based on the lesions and history we concluded that the ruminal bloat was due to a development failure of the omasum...

Multiple effects of bracken fern under in vivo and in vitro conditions.

Several toxic substances have been detected in plants which are responsible for animal and human diseases. Bracken fern (Pteridium aquilinum) is one example, widely distributed in many parts of the world. It is known to cause cancer in humans and other animals. In fact, man can be directly or indirectly exposed to the danger by consuming fern, contaminated water, milk, meat, and spore inhalation. Experimental studies have shown an association between bracken exposure and gastric cancer, and research has shown genotoxic and cytotoxic effects in vitro. This paper describes and reviews toxic, carcinogenic, genotoxic/cytotoxic, and immunomodulatory effects of bracken and included possible toxic agents. The chemistry of Ptaquiloside (PT) reactions is emphasized, along with bracken problems in livestock, possible pathways of exposure in man, and control for human health.

Hematuria and urinary bladder lesions compatible with bracken fern (Pteridium aquilinum) intoxication in captive fallow deer (Dama dama).

Hemangiosarcoma, hemangioma, transitional cell carcinoma, and chronic cystitis were diagnosed in the urinary bladder of six captive fallow deer (Dama dama). Hematuria and thin body condition were observed in the advanced cases. These findings were compatible with chronic enzootic hematuria and were suspected to have been induced by chronic ingestion of bracken fern (Pteridium aquilinum) present on the premises. These lesions were similar to those described in bovine enzootic hematuria in cattle. Hemangiosarcoma metastases, hydronephrosis, and renal carcinoma were also associated in some cases. This is the first report of hemangioma, hemangiosarcoma, transitional cell carcinoma, and renal carcinoma of the urinary bladder in fallow deer and the first indication of bracken fern intoxication in deer.

Importância econômica, epidemiologia e controle das intoxicações por plantas no Brasil / Economic impact, epidemiology and control poisonous plants in Brazil

A importância econômica, epidemiologia e controle das intoxicações por plantas em animais domésticos no Brasil são revisadas. Com os dados dos laboratórios de diagnóstico de diferentes regiões do país, as perdas anuais por mortes de animais foram estimadas em 820.761 a 1.755.763 bovinos, 399.800 a 445.309 ovinos, 52.675 a 63.292 caprinos e 38.559 equinos. No Brasil, atualmente, o número de plantas tóxicas é de 131 espécies e 79 gêneros e aumenta permanentemente. No entanto, a maioria das perdas são causadas por poucas plantas, incluindo Palicourea marcgravii, Amorimia spp., Senecio spp., Pteridium aquilinum, Ateleia glazioviana e Cestrum laevigatum em bovinos, Brachiaria spp em bovinos e ovinos, Nierembergia veitchii, Mimosa tenuiflora e Ipomoea asarifolia em ovinos, plantas que contêm swainsonina (Ipomoea carnea, Turbina cordata e Sida carpinifolia) em caprinos e Brachiaria humidicola e Crotalaria retusa em equinos. Os principais fatores epidemiológicos relacionados às intoxicações por plantas incluem palatabilidade, fome, sede, facilitação social, desconhecimento da planta, acesso a plantas tóxicas, dose tóxica, período de ingestão, variações de toxicidade e resistência/susceptibilidade dos animais às intoxicações. Quanto aos métodos de controle e profilaxia descrevem-se os resultados obtidos no Brasil com métodos recentemente desenvolvidos, incluindo controle biológico, aversão alimentar condicionada, utilização de variedades não tóxicas de forrageiras, utilização de animais resistentes às intoxicações e técnicas de indução de resistência.

The economic impact, epidemiology and control of plant poisonings in Brazil are reviewed. With the data obtained from diagnostic laboratories from different Brazilian regions, annual losses due to animal deaths are estimated in approximately 820.761 to 1.755.763 cattle, 399.800 to 445.309 sheep, 52.675 to 63.292 goats, and 38.559 horses. In Brazil toxic plants include 131 species within 79 genera, but this number is increasing continuously. However, most economic losses are caused by few plants including Palicourea marcgravii, Amorimia spp., Senecio spp., Pteridium aquilinum, Ateleia glazioviana and Cestrum laevigatum in cattle, Brachiaria spp. in cattle and sheep, Nierembergia veitchii, Mimosa tenuiflora and Ipomoea asarifolia in sheep, swainsonine-containing plants (Ipomoea carnea, Turbina cordata and Sida carpinifolia) in goats, and Brachiaria humidicola and Crotalaria retusa in horses. The main factors that determine the occurrence and frequency of plant poisoning include palatability, hunger, thirst, social facilitation, unawareness of the plant, access to toxic plants, toxic dose, ingestion period, variations in toxicity, and animal resistance/susceptibility. The results obtained in Brazil with the use of biological control, conditioned food aversion, selection of non-toxic varieties of forages, use of resistant animals to poisoning, and techniques to induce resistance are described.

Sobrevivência/viabilidade de bovinos com Hematúria Enzoótica após transferência para região livre de Pteridium arachnoideum / Survival/viability of cattle with Bovine Enzootic Hematuria after transfer to area free of Pteridium arachnoideum

With the objective of explaining the question of survival of cattle affected by Enzootic Hematuria, when transferred to areas free of Pteridium arachnoideum, the study was accomplished in two parts: a) by a questionnaire regarding the epidemic aspects of that illness, which was answered by the proprietors of 73 cattle establishments visited in the areas of Southeast of Brazil, where P. arachnoideum is prevalent. The applied questionnaire to the proprietors indicates that enzootic hematuria is responsible for serious socioeconomic problems; b) clinical and laboratorial attendance of 51 cattle of this area affected by Enzootic Hematuria, between 2007 and 2011, transferred to area free of P. arachnoideum. More than 90% of the affected animals by HEB died before two years after removal from areas free of P. arachnoideum.

Com o objetivo de esclarecer a questão da sobrevida de bovinos afetados por Hematúria Enzoótica, quando transferidos para áreas livres de Pteridium arachnoideum, o estudo foi realizado em duas partes: a) aplicação de questionário versando sobre os aspectos epidemiológicos dessa enfermidade, respondido por proprietários de 73 estabelecimentos pecuários visitados nas áreas da Região Sudeste onde P. arachnoideum é prevalente. O questionário aplicado aos proprietários indica que a HEB é causa de sérios problemas sócio-econômicos. b) acompanhamento clínico e laboratorial de 51 bovinos desta região, afetados pela Hematúria Enzoótica entre 2007 e 2011, transferidos para região livre de P. arachnoideum. Verificou-se que mais de 90% dos animais afetados por HEB morre antes de dois anos após serem transferidos para áreas indenes de P. arachnoideum.

Bracken-fern extracts induce cell cycle arrest and apoptosis in certain cancer cell lines.

Bracken fern [Pteridium aquilinem (L.) kuhn (Dennstaedtiaceae)] is one of the most common species on the planet. It has been consumed by humans and animals for centuries. Use by some human groups is because they believe bracken fern is good for health as plant medicine. However, it is also one of the few known plants that can cause tumors in farm animals. Many interested groups have focused their attention on bracken fern because of these interesting features. In order to evaluate the biological effects of exposure to this plant in cellular level, human cancer cell lines were treated with the fern dichloromethane extracts and the genotoxic and cytotoxic effects were studied. Anti-proliferative/cytotoxic effects were evaluated by cell count, MTT assay and flow cytometry methods with three different cancer cell lines, TCC, NTERA2, and MCF-7, and two normal cells, HDF1 and HFF3. Pro-apoptotic effects of the extracts were determined by DAPI staining and comet assay, on TCC cancer cells compared to the normal control cell lines. Cellular morphology was examined by light microscopy. Our present study showed that the extract caused DNA damage and apoptosis at high concentrations (200 µg/mL) and also it may induce cell cycle arrest (G2/M phase) at mild concentrations (50 and 30 µg/mL) depending on the cell type and tumor origin. These results indicate that bracken fern extract is a potent source of anticancer compounds that could be utilized pharmaceutically.

Pteridium aquilinum: o que sabemos e o que ainda falta saber / Pteridium aquilinum: what we know and what is yet to be learnt

A samambaia Pteridium aquilinum encontra-se amplamente distribuída no planeta, sendo considerada um problema ambiental, além de representar perigo para a saúde tanto de animais quanto da espécie humana. A taxonomia do gênero Pteridium tem sido objeto de discussão e novas propostas de classificação elevam variedades à categoria de espécies ou subespécies. Ao longo da história muito já se aprendeu sobre suas características biológicas, o que permitiu a adoção de medidas de prevenção de sua habilidade invasora e de seus efeitos tóxicos. Não obstante, ainda há aspectos a serem esclarecidos sobre como tais efeitos são exercidos, sobre seu(s) princípio(s) ativo(s) e respectivo mecanismo de ação, com o ptaquilosídeo sendo o composto mais estudado. Importantes efeitos desta substância foram demonstrados no nível molecular. Estudos epidemiológicos realizados com consumidores do vegetal revelaram elevação do risco para câncer de esôfago e estômago em associação à sua ingestão.

Bracken fern (Pteridium aquilinum) is present in practically all parts of Earth's surface, being considered a serious environmental problem besides representing a hazard for both animals and human beings. The taxonomy of the genus Pteridium has been discussed and former varieties are now proposed to be considered as species or subspecies. Along history much has been learnt about its biological characteristics, what has allowed the adoption of preventive measures against its encroaching ability and its toxic effects. Nevertheless there are still aspects to be clarified about how these effects are exerted, about its active principle(s) and respective mechanism of action, with ptaquiloside being the most studied compound. Important effects of this substance have been demonstrated at the molecular level. Epidemiological studies performed with consumers of the plant revealed an increase of the risk for esophageal and stomach cancer associated with its ingestion

Intoxicação experimental aguda por samambaia (Pteridium aquilinum) em bovinos / Experimental acute poisoning by bracken fern (Pteridium aquilinum) in cattle

Este experimento foi delineado para investigar os seguintes pontos em relação à intoxicação aguda por samambaia (Pteridium aquilinum) em bovinos: 1) a intensidade da trombocitopenia em diferentes momentos da intoxicação e sua relação com possíveis déficits na hemostasia secundária, 2) a relação da neutropenia com as manifestações morfológicas de septicemia ocasionalmente observadas na necropsia, e 3) o mecanismo da anemia e sua relação com a perda de sangue, a vida média eritróide e a evolução da doença. As hastes superiores mais verdes de P. aquilinum foram administradas a quatro bovinos sem raça definida, com idade média de 1,5 ano e pesos entre 190-215 kg. Um bovino de idade e peso semelhantes foi usado como controle e, exceto por não ter recebido P. aquilinum, foi mantido nas mesmas condições que os outros quatro. Os quatro bovinos que receberam a planta morreram com quadro característico da intoxicação aguda por samambaia após receberem durante 53-58 dias, doses diárias de 8,0, 8,6, 10,2 e 10,6g/kg de peso corporal, que totalizaram, ao final do experimento, respectivamente, 112,7, 107,6, 85,7, 90,15 kg da planta, o que corresponde, respectivamente, a 59,3 por cento, 63,3 por cento, 47,4 por cento, 47,5 por cento da planta em relação ao peso dos bovinos. A doença caracterizou-se por febre de até 42,5 graus C e diversos graus de hemorragias observadas clinicamente, na necropsia e na histopatologia. A morte ocorria 6-7 dias após o início do quadro febril. As alterações hematológicas revelaram trombocitopenia e neutropenia acentuadas. Em dois dos quatro bovinos havia anemia leve. Não houve variações significativas nos tempos de coagulação dos bovinos intoxicados, quando avaliados os fatores de coagulação (secundária), excluindo-se assim a possibilidade da participação de distúrbios da hemostasia secundária na patogênese das hemorragias nessa intoxicação. A determinação dos produtos da degradação da fibrina no soro revelou dados conflitantes...

This experiment was design to address the following points in relation to the acute poisoning by bracken fern (Pteridium aquilinum) in cattle: 1) the severity of the thrombocytopenia in different stages of the intoxication and its relationship to possible deficits in the secondary hemostasis, 2) the relationship between neutropenia and the morphological signs of septicemia occasionally found at necropsy, and 3) the mechanism of anemia and its relationship with blood loss, medium life of erythrocytes and the progress of the disease. The fresh green upper parts of P. aquilinum were fed to four mixed breed calves with average age of 18 months and weights ranging from 190 to 215 kg. A calf of similar age and weight was kept together with the other four under the same conditions, except for the ingestion of P. aquilinum. The four fern-fed calves died with typical features of acute bracken fern poisoning after being fed with the plant for 53-58 days daily doses of 8.0, 8.6, 10.2, and 10.6g/kg body weight totaling at the end of the experiment, respectively, 59.3 percent, 63.3 percent, 47.4 percent, and 47.5 percent of bracken fern in relation to their body weight. The disease was characterized by fever up to 42.5 grades C and varying degrees of hemorrhages observed clinically, at necropsy and on histological examination. Death occurred 6-7 days after the onset of fever. The hematological changes consisted mainly of marked thrombocytopenia and neutropenia. Two of the four affected calves had mild anemia. The deficits in secondary hemostasis were mild in each case. There were no significant changes thus permitting to rule out the role of secondary hemostasis in the pathogenesis of the hemorrhages in the bracken fern poisoning. The measurement of fibrin degradation products in the serum showed conflicting results and did no allow for a solid conclusion regarding the role of disseminated intravascular coagulation in the pathogenesis of the hemorrhages...

A biological hazard of our age: bracken fern [Pteridium aquilinum (L.) Kuhn]--a review.

Bracken fern (Pteridium aquilinum) is the fifth most distributed common weed species of the world. Its ecological distribution is very wide, and the plant can grow and spread successfully on many types of soil. The cover of P. aquilinum is--in some cases--remarkable (e.g., in the United Kingdom). Bracken fern contains different poisonous agents: some cyanogen glycosides, factors (agents) of antithiamine character (thermolabile thiaminase and thermostable other compounds) and factors of carcinogenic activity (first of all ptaquiloside). This paper summarises and reviews different toxicological problems and poisonings caused by bracken fern in ruminants (cattle, sheep) and in non-ruminant animals (horses, pigs, rats, mice, etc.). The carcinogenic properties of the norsesquiterpene-type ptaquiloside make bracken fern a potent, living hazard. Recent investigations have shown that ptaquiloside pollution of different soil layers is a distinct possibility. Ptaquiloside may leach from the soil into the drinking water base. This ecotoxicological aspect seems to be the most hazardous phenomenon in relation to P. aquilinum and ptaquiloside. The carcinogenic effect of ptaquiloside is based on its hydrolysis, which leads to the formation of a dienon intermediate. It can produce DNA adducts, which are responsible for inducing carcinoma.

DNA damage and apoptosis induced by Pteridium aquilinum aqueous extract in the oral cell lines HSG and OSCC-3.

BACKGROUND: Bracken fern (Pteridium aquilinum) has been consumed by humans and animals for centuries. However, its consumption is associated with a high incidence of cancer in the upper digestory tract of different species. Although the oral cavity is the first site of contact with ingested toxic substances, the interaction of bracken fern composites with oral cell lines has not yet been studied. METHODS: In order to study the biological responses of oral cells exposed to bracken fern, we evaluated the genotoxic and cytotoxic effects of a bracken fern aqueous extract in oral cell lines. Human submandibular gland (HSG) and human oral epithelium cells (OSCC-3) cells were treated with three different concentrations of the extract. DNA damage was determined by the comet assay, and cellular morphology was examined by light microscopy. Apoptotic changes were evaluated by transmission electron microscopy and TUNEL assay. RESULTS: The comet assay revealed that the extract was genotoxic for both cell lines but the results were not dose-dependent. The morphological and ultrastructural analyses showed that the extract caused conspicuous alterations in both cell types: uncommon chromatin condensation, nuclear picnosis, cellular volume decrease, nuclear envelope disruption, formation of numerous vacuoles of different sizes and apoptotic bodies. The TUNEL assay confirmed apoptosis induction. CONCLUSIONS: These results demonstrate that the extract was cytotoxic to HSG and OSCC-3 cells, and that cellular degeneration occurred mainly by apoptosis. We believe that oral cells could trigger apoptosis after bracken fern induced DNA damage, in order to avoid the malignant transformation.

Intoxicação aguda por samambaia (Pteridium aquilinum) em bovinos na Região Central do Rio Grande do Sul / Acute poisoning by bracken fern (Pteridium aquilinum) in cattle in central Rio Grande do Sul, Brazil

Foram revisados casos de intoxicação por samambaia (Pteridium aquilinum) em bovinos de 10 pequenas propriedades rurais de sete municípios da Região Central do Rio Grande do Sul. O estudo abrangeu 6.256 necrospias de bovinos realizadas num período de 43 anos e incluiu 15 necropsias de bovinos que morreram em conseqüência da intoxicação aguda por samambaia. As taxas de morbidade e mortalidade foram 17,9 por cento e a letalidade era virtualmente de 100 por cento. Em 40 por cento das propriedades a doença ocorreu em pequenos surtos afetando vários bovinos; em 60 por cento das propriedades apenas um bovino era afetado. Os principais sinais clínicos incluíam febre (40-42ºC), apatia, salivação e hemorragias, principalmente pelas gengivas, pela cavidade nasal e pelo trato gastrintestinal. Múltiplas petéquias eram observadas nas mucosas e na pele. Ocasionalmente observou-se hematúria e sangue no leite. A doença era invariavelmente fatal após um curso clínico de dois dias. Alterações hematológicas consistiam de leucopenia por neutropenia, anemia normocítica normocrômica e trombocitopenia arregenerativa. Os achados de necropsia incluíam hemorragias de intensidade variável em vários órgãos e infartos no fígado. Tanto as hemorragias quanto os infartos foram confirmados histologicamente; agregados de bacilos e vasos trombosados foram observados em associação com os infartos. Aplasia da medula óssea era um achado consistente nos quatro casos em que esse órgão foi examinado.(AU)

Cases of acute poisoning by bracken fern (Pteridium aquilinum) in cattle from 10 small farms of seven municipalities of the Central Region of Rio Grande do Sul State, Brazil, were reviewed. The study encompassed 6,256 necropsies of cattle considering a 43-year period and included 15 necropsies of cattle dying from acute bracken poisoning. Morbidity and mortality were 17.9 percent and lethality was virtually 100 percent. In 40 percent of the farms the disease occurred in small outbreaks affecting several cattle per farm and in 60 percent only one bovine was affected in each farm. Main clinical signs include fever (40-42ºC), apathy, drooling, and hemorrhages; the latter were mainly from the gums, nostrils, and gastrointestinal tract. Multiple petechiae were observed in several mucosae and in the skin. Occasionally hematuria and blood in the milk were reported. The disease was invariably fatal after a course of approximately two days. Hematological changes included severe neutropenia, non-regenerative normocytic normochromic anemia and thrombocytopenia. Necropsy findings include hemorrhages o varying degrees in several organs and infarcts in the liver. Both hemorrhages and hepatic infarcts were confirmed histologically; clusters of bacterial rods and thrombosed vessels were associated with the infarcts. Marked bone marrow aplasia was a consistent finding in the four cases in which the marrow was evaluated.(AU)

Clinico-pathological findings in cattle exposed to chronic bracken fern toxicity.

AIM: To describe changes in blood and urine analytes in a large group of cattle exposed to chronic bracken fern toxicity, in order to identify parameters of potential diagnostic value. METHODS: The study was conducted on two livestock farms on which bovine enzootic haematuria (BEH) was known to occur; Farm A grazed a local breed of cows and Farm B grazed Friesians. Group A1 comprised 66 cows from Farm A, Group B 54 cows from Farm B, and Group A2 13 heifers from Farm A. Ten healthy cows were used as controls. A complete physical examination was performed (Group A1), and blood (all groups) and urine (Groups A1 and B) samples were collected. Necropsies and histopathology were undertaken on four cows. RESULTS: Anaemia, leucopenia, monocytosis, thrombocytopenia, hypergammaglobulinaemia, microhaematuria and proteinuria were detected. Multivariate statistical analyses established three phases of the disease of increasing severity; an initial phase, characterised by an extremely high monocytosis and otherwise normal parameters; an intermediate phase, characterised by monocytosis and moderate changes to other analytes; and a final phase, characterised by normal levels of monocytes and many changes to other analytes. CONCLUSIONS AND CLINICAL RELEVANCE: Monocytosis, detected in 31% of the younger animals, could represent an initial response to consumption of bracken fern and might be useful as an early haematological marker of BEH.

Urinary bladder lesions in bovine enzootic haematuria.

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 São 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.

Alterações anátomo-histopatológicas em ratos submetidos à dieta com extrato de broto de samambaia (Pteridium aquilinum var. arachnoideum) de dois municípios do estado do Paraná / Macroscopic and microscopic changes in rats fed on a bracken fern diet (Pteridium aquilinum var. arachnoideum) from different regions of Parana State, Brazil

The carcinogenicity of bracken fern harvest from two regions of Paraná State to induce hematury in rats was studied. In order to do that, 33 Wistar rats were divided in three groups. Groups I and II received an aqueous extract of bracken fern from Londrina-PR or Ibaiti-PR, respectively, in drinking water for 60 days. Group III, control group, received regular plain water with no bracken fern. After 15 months, euthanasia was performed in all animals and samples were collected for histology examination. Histologic analysis revealed that two animals of Group II had ileal adenocarcinoma and soft tissue fibroma of leg.

H-ras immunohistochemical expression and molecular analysis of urinary bladder lesions in grazing adult cattle exposed to bracken fern.

Chronic ingestion of bracken fern (Pteridium spp.) by cattle produces upper alimentary tract and urinary bladder tumours causing a syndrome called bovine enzootic haematuria (BEH). Previous studies demonstrated ptaquiloside-DNA adducts and mutations in the h-ras gene in ileal epithelial cells of bracken fern-fed calves. Systematic inspection of the bladder mucosa of grazing cattle (n=126) from bracken-fern areas was carried out in a slaughterhouse. Of the 126 slaughterhouse cattle, 46 showed macroscopical lesions of the bladder. These bladders, together with six others known to have BEH, were examined histopathologically and by H-ras immunohistochemistry. Thirteen affected bladders were also examined by H-ras molecular analysis to detect mutations. Macroscopical and histological study of urinary bladder lesions found at the slaughterhouse revealed chronic cystitis (34.1%) and tumours (2.4%). There was significantly increased immunohistochemical expression of H-ras (P<0.05) in chronic cystitis (H-ras=53.24%) and bladder tumours (H-ras=63.60%) as compared with normal urinary bladders (H-ras=4.32%). A silent mutation (D38D) was detected in one animal with a mixed bladder tumour. The prevalence of urinary bladder lesions (chronic cystitis and tumours) obtained at the slaughterhouse was higher than expected. This study demonstrates that close inspection of urinary bladders of adult grazing cows is necessary to prevent possible human exposure to bracken-fern carcinogens. The absence of mutations in the codons of h-ras studied did not exclude the presence of polymorphisms in other regions of the gene (promoter or regulation sequences) or in other genes (belonging or not to the ras family) that significantly affect the H-ras protein.