Comparison of two collection methods to determine protodioscin concentrations in Brachiaria spp. and to monitor Brachiaria spp. poisoning / Comparação entre dois métodos de coleta para determinar as concentrações de protodioscina em Brachiaria spp. e monitorar a intoxicação por Brachiaria spp

The objective of this study was to evaluate the methods of collecting Brachiaria spp. (Simulated Grazing vs. Total Collection) for the determination of protodioscin concentrations and to monitor the intoxication in sheep susceptible and resistant to poisoning by Brachiaria spp. The collection of Brachiaria spp. was done through two different methods. The first one consisted in throwing a metallic square at 10 random picket points. The second was performed simulating, with the hands, the grazing behavior of sheep. The leaf blades obtained by the two methods were conditioned in a forced ventilation oven. The extraction and quantification of protodioscin from the dry milled plant material was done by reverse-phase high-performance liquid chromatography (HPLC). The collections were carried out at three stages of the life of experimental lamb (suckling, rearing, and finishing). The lambs were divided into resistant and susceptible to intoxication by Brachiaria spp. The criterion for a lamb to be considered sensitive was to present some clinical signs at some stage of its life. No significant differences were observed between the general averages of the two collections when the evaluation was done in the whole period. However, a significant difference was found at the onset of the clinical signs. The concentrations of protodioscin in this period were higher in Brachiaria spp. collected by the simulating grazing method. Both methodologies were efficient to determine the level of protodioscin in the pastures of Brachiaria spp. However, during the periods when the outbreaks occurred, the simulated grazing method was more reliable.(AU)

Objetivou-se avaliar dois métodos de coleta de Brachiaria spp. (Pastejo Simulado vs Coleta Total) para determinação das concentrações de protodioscina e efetuar o monitoramento da intoxicação em rebanhos sensíveis e resistentes à intoxicação por Brachiaria spp. Foram realizadas coletas de Brachiaria spp. através de duas metodologias. A primeira consistiu em fazer o lançamento de um quadrado metálico em 10 pontos aleatórios do piquete. A segunda foi realizada simulando com as mãos o comportamento de pastejo dos ovinos. As lâminas foliares obtidas pelas duas metodologias foram acondicionadas em estufa de ventilação forçada. O material seco e moído foi encaminhado para extração e quantificação da protodioscina por cromatografia líquida de alto desempenho (HPLC) de fase reversa. As coletas foram realizadas em três fases de vida dos cordeiros (cria, recria e terminação). Os cordeiros foram divididos em resistentes e sensíveis a intoxicação por Brachiaria spp. O critério para o ovino ser considerado sensível foi apresentar algum sinal clínico da intoxicação em alguma fase de sua vida. Não foram observadas diferenças significativas entre a média geral das duas coletas quando a avaliação foi feita no período total. Porém, foi observada diferença significativa no período de aparecimento dos casos. As concentrações de protodioscina neste período foram superiores pelo método do pastejo simulado. Ambas as metodologias mostraram-se eficientes para determinação da concentração da protodioscina nas pastagens de Brachiaria spp. No entanto, nos períodos em que ocorreram os surtos, o método do pastejo simulado foi mais fidedigno.(AU)

Identification of lamb flocks susceptible and resistant against Brachiaria poisoning / Identificação de rebanhos de cordeiros suscetíveis e resistentes a intoxicação por braquiária

This study was designed to assess the influence of genetic resistance against brachiaria poisoning in sheep. Two groups of sheep, one identified as susceptible (formed by two ewes and one ram) and the other as resistant against brachiaria poisoning (formed by three ewes and one ram) were selected. Sheep considered susceptible were those that presented clinical signs of brachiaria poisoning at any time of their life; resistant sheep were those that even raised on Brachiaria spp. pastures, did not developed any sign of the poisoning during their life. The offspring of the two flocks (15 lambs from the sensitive flock and 9 lambs from the resistant flock) were placed into brachiaria pasture (initially Brachiaria decumbens and B. brizantha,and only B. decumbens after weaning) and followed up during two years (2013-2014). The determination of protodioscin levels in B. decumbens pasture was performed only in 2014 and revealed significant amounts of the toxic principle. Eleven lambs of the susceptible group were affected to some degree of brachiaria poisoning and six died; no lamb of the resistant group was affected. Clinical signs consisted of varying degrees of subcutaneous edema of the face and, erythema and loss of hair of the ears, crusts on the skin of ears, around the eyes and on planum nasale, scar deformation of the ears, and bilateral ocular discharge; affected lambs also sought for shadowy shelters and they were poor doers. Several sheep recovered from the condition and then relapsed. Necropsy findings in six lambs included pale mucous membranes, emaciation, dermatitis, scar deformation of the ears, large yellow livers with marked lobular pattern, and moderate infestation by Haemonchus contortus...

Este estudo avaliou a resistência genética na ocorrência de intoxicação por braquiária em ovinos. Foram selecionados dois grupos de ovinos, um identificado como suscetível (formado por duas ovelhas e um carneiro) e o outro como resistente (formado por três ovelhas e um carneiro). Foram considerados suscetíveis ovinos que apresentaram sinais de intoxicação por Brachiaria spp. em algum ponto de suas vidas e resistentes aqueles ovinos que, mesmo criados em pastagem de braquiária, nunca desenvolveram qualquer sinal da intoxicação. A progênie desses dois grupos (15 cordeiros do grupo suscetível e 9 no grupo resistente) foi colocada numa pastagem de braquiária (inicialmente Brachiaria decumbens e B. brizantha e, após o desmame, apenas B. decumbens) e acompanhada durante dois anos (2013-2014). A determinação dos níveis de protodioscina em B. decumbens foi realizada apenas em 2014 e foram encontradas quantidades significativas do princípio tóxico. Onze cordeiros do grupo suscetível foram afetados por algum grau de intoxicação por braquiária; nenhum cordeiro do grupo resistente foi afetado. Os sinais clínicos consistiam de graus variáveis de edema subcutâneo da face e eritema e alopecia da pele das orelhas, crostas na pele das orelhas e ao redor dos olhos e no plano nasal, retração cicatricial das orelhas, fotofobia e corrimento ocular bilateral. Três cordeiros apresentaram desenvolvimento retardado. Vários cordeiros se recuperaram da condição, mas posteriormente quando foram colocados na pastagem apresentaram recidivas. Achados de necropsia em seis cordeiros incluíam mucosas pálidas, pobre condição corporal, dermatite, deformação cicatricial das orelhas, fígado aumentado de volume, amarelo e com padrão lobular evidenciado e graus moderados de infestação por Haemonchus contortus...

Spontaneous poisoning by brachiaria decumbens in goats / Intoxicação espontânea por brachiaria decumbens em caprinos

An outbreak of Brachiaria decumbens poisoning in goats in the state of Mato Grosso do Sul, Brazil, is described. Out of a herd of about 1,000 goats, seven goats died after showing photosensitivity (3 out 7 goats) and icterus (4/7). Other clinical signs included lethargy, dehydration and weight loss (7/7). At necropsy there was either hepatomegaly or liver atrophy; orange, yellow or gray discoloration - were observed in the liver of six goats. Two had accentuation of lobular pattern and punctate, linear or stellar fibrous depressions were randomly distributedover the capsular surface. Macroscopic changesattributedto liver failure consisting of hydropericardium (2/7) and ascites (2/7) were observed. Histologically, there was accumulation of refringent crystals in the bile ducts and infiltration of foamy macrophages in the liver of all goats. Foamy macrophages were also present in the hepatic and mesenteric lymph nodes of two goats. The diagnosis of B. decumbens poisoning was based on epidemiology, clinical signs, necropsy findings and histopathology.

Relata-se um surto de intoxicação por Brachiaria decumbens em caprinos no estado do Mato Grosso do Sul. De um rebanho de aproximadamente 1.000 caprinos, sete cabras morreram após apresentar fotossensibilização (3 de 7 cabras) e icterícia (4/7). Outros sinais clínicos incluíam apatia, desidratação e emagrecimento (7/7). Todas as cabras tinham alteração no tamanho do fígado - cinco estavam aumentados de volume e dois, diminuídos; o fígado de 6 cabras tinha alteração na cor - dois eram alaranjados, dois amarelados e dois, acinzentados; o fígado de duas cabras tinha acentuação do padrão lobular e outros dois apresentavam depressões fibrosas puntiformes, lineares ou estreladas, brancacentas distribuídas aleatoriamente pela superfície capsular. Adicionalmente, foram observadas alterações macroscópicas atribuídas à insuficiência hepática como hidropericárdio (2/7) e ascite (2/7). Alterações histopatológicas incluíam cristais refringentes nos ductos biliares e infiltrado de macrófagos espumosos no parênquima hepático de todas as cabras. Macrófagos espumosos também foram observados nos linfonodos mesentéricos e hepáticos em duas cabras. O diagnóstico de intoxicação por braquiária foi baseado na epidemiologia, achados clínicos, de necropsia e histopatológicos.

Extraction of saponins and toxicological profile of Teucrium stocksianum boiss extracts collected from District Swat, Pakistan

BACKGROUND: The current era is facing challenges in the management of neoplasia and weeds control. The currently available anti-cancer and herbicidal drugs are associated with some serious side effects. Therefore numerous researchers are trying to discover and develop plant based alternative particularly for the rational management of cancer and weed control. Teucrium stocksianum possess antioxidant and analgesic activities. The current study was designed to evaluate crude saponins (CS), methanolic extract and sub-fractions of T. stocksianum for cytotoxic and phytotoxic potentials. CS, methanolic extract and sub-fractions were extracted from powdered plant material using different solvents. Cytotoxic potential of the extracts at a dose of 10, 100 and 1000 µg/ml were evaluated against Brine shrimp's nauplii. Phytotoxic assay also performed at the same concentration against Lemna minor. Etoposide and Paraquat were used as positive controls in cytotoxic and phytotoxic assays respectively. RESULTS: The percent yield of crude saponins was (5%). CS demonstrated tremendous brine shrimp lethality showing < 10 µg/ml LC50. The n-hexane (HF) and chloroform fractions (CF) demonstrated excellent cytotoxicity with 80 and 55 µg/ml LC50 respectively. Whereas the methanolic extract (TSME), ethyl acetate (EAF) and aqueous fractions (AF) revealed moderate cytotoxicity showing 620, 860 and 1000 µg/ml LC50 values respectively. In phytotoxic assay profound inhibition was displayed by HF (96.67%) and TSME (95.56%, 30 µg/ml LC50) against the growth of Lemna minor at 1000 µg/ml respectively. Both CF and EAF demonstrated profound phytoxicity (93.33%) respectively at highest concentration (1000 µg/ml), while AF and CS demonstrated weak phytotoxicity with 1350 and 710 µg/ml LC50 values respectively. CONCLUSION: Cytotoxicity and phytotoxicity assays indicated that the crude saponins, n-hexane and chloroform fractions of T. stocksianum could play a vital role in the treatment of neoplasia and as potential natural herbicides. Therefore these sub-fractions are recommended for further investigation with the aim to isolate novel anti-cancer and herbicidal compounds.

Steroidal glycosides of furcraea selloea and their biological properties against diffrent schistosoma mansoni stages

Furcraea selloea C. Koch dry powder [Family Agavaceae] was subjected to a bioassayguided fractionation technique to isolate the active constituents responsible for the potency of this plant. The antischistosomal impact of different extracts of the leaves of F. selloea was screened against adult Schistosoma mansoni warms in vitro using a well established culture media. The methanol extract of the plant showed the highest activity as S. mansoni warms recorded 100% mortality at 50 micro g/ ml after 24 hours. Owing to the high potency of the crude saponins obtained from the methanolic extract [100% mortality at 20 micro g/ml], it was submitted to chromatographic separation using silica gel and Sephadex columns as well as preparative thin layer chromatography. Three steroidal saponins [I-III] were isolated and their structures were elucidated using some spectroscopic and chemical methods as follows: 6-O-Beta-D- glucopyranosyl [1-4]-Beta-D-glucopyranoside chlorogenin [I], 3-O-Beta-D-glucopyranosyl-[1-4]- Beta-D-glucopyranoside crestagenin [II] and 3-O-Beta-D-glucopyranosyl-[1-3]-Beta-D- glucopyranosyl-[1-3]-Beta-D-xylopyranoside gloriogenin [III]. Bioassay screening of the isolated saponins [I-III] were carried out against certain Schistosoma mansoni stages. Compound III only proved to possess antischistosomal activity against S. mansoni worms at concentration as low as 5 micro g/ ml, while compounds I and II were inactive. Also, test against B. alexandrina snails revealed that only saponin III has high molluscicidal activity [LC [90] = 6 ppm] whereas the other two saponins did not show any activity up to 50 ppm after 24 hours exposure. Different concentrations of the crude and the isolated saponins were evaluated against S. mansoni free larval stages [cercariae and miracidia]. Hatchability of S. mansoni ova was markedly depressed when exposed to 6 ppm of compound III. The infection rate of B. alexandrina snails was significantly reduced when snails were exposed to three sublethal concentrations of the dry plant powder. Determination of the acute oral toxicity of F. seiloea methanol extract against mice was carried out. When three groups of mice infected with S. marisoni were treated orally with a single dose of 2500 mg/kg of F. selloea methanol extract either at 2, 4 or 7 weeks post infection, the reduction rate in worm load was significantly lower when compared to infected untreated control