Accidental Poisoning with Nerium Oleander in Dairy Goat in the Santiago Metropolitan Region in Chile / Intoxicación accidental con Nerium oleander en lechería caprina en la Región Metropolitana de Santiago en Chile / Intoxicação acidental com oleandro de Nerium em laticínios de cabras na região metropolitana de Santiago, no ChileResumo

Abstract A small dairy goat farm in central Chile stocked their troughs with ornamental plants and grasses, which were obtained after pruning the gardens of neighboring sectors. Twelve hours after the goats ingested this food, the farm registered a death toll of eight goats, all of which were adult females in their productive stage, before entering the milking parlor. The mortality episode lasted 84 hours and produced mortality and morbidity values of 48 and 54 percent, respectively (n = 52). During a visit to the farm, clinic inspections, autopsies, and sample tissues were obtained for microscopic analysis (the tissues specimens were fixed in 10% buffered formaldehyde solution). The clinic inspection and microscopic analysis both confirmed that the animals had cardiovascular disease. The lesions were compatible with those produced by cardiac glycosides. During the site visit, Nerium oleander was found in the troughs, which suggested that the animals were poisoned by eating this plant. There was a high presence of this plant in the poisoned area, and it was not associated with the mortality episode before the on-site visit, which indicates that the authors should conduct training and inform producers about the handling and feeding of their animals.

Resumen En establecimiento de pequeño productor caprino (n = 52) en la zona central de Chile, se les administraron en comederos plantas ornamentales y pastos que fueron obtenidos posterior a la poda de jardines en sectores aledaños. A las 12 horas post ingesta de este alimento se encuentran 8 caprinos, hembras, adultos, productivos muertos previo al ingreso a sala de ordeña. El episodio de mortandad se extendió por 84 horas generando una mortalidad del 48 % y una morbilidad del 52 %. Se realizó visita, inspección clínica, necropsia y análisis microscópico de especímenes fijados en solución formolada tamponada al 10 %. La inspección clínica confirma la presencia de animales con afección cardiovascular, al igual que el análisis microscópico. Las lesiones son compatibles con las producidas por glucósidos cardiotóxicos, al recorrer el lugar se observa la presencia de Nerium oleander en comederos de lote afectado, lo que sugiere la intoxicación de estos animales. La presencia de esta planta en la zona de la intoxicación es elevada y antes de nuestra visita no fue asociada al episodio de mortandad, lo que nos indica que se debe realizar capacitación e informar a productores sobre el manejo y alimentación de sus animales.

Resumo No estabelecimento de um pequeno produtor de cabras (n = 52) na zona central do Chile, plantas ornamentais e pastagens foram administradas em alimentadores que foram posteriormente podados por jardins em setores próximos. Às 12 horas após a ingestão deste alimento, são oito cabras, fêmeas, adultos, mortos produtivos antes de entrar na sala de ordenha. O episódio da morte prolongou-se por 84 horas, gerando uma mortalidade de 48% e uma morbidade de 52%. Foram realizadas visita, inspeção clínica, necropsia e análise microscópica das amostras fixadas em solução formolada tamponada a 10%. A inspeção clínica confirmou a presença de animais com doença cardiovascular e a análise microscópica. As lesões são compatíveis com as produzidas pelos glicosídeos cardiotóxicos, ao atravessar o local, observa-se a presença de oleandro de Nerium nos alimentadores do lote afetado, o que implica na intoxicação desses animais. A presença desta planta na zona de envenenamento é alta e, antes da nossa visita, não estava associada ao episódio da morte, o que indica que o treinamento deve ser realizado e informa os produtores sobre o manuseio e a alimentação de seus animais.

Improved method for diagnosis of Nerium oleander poisoning in necropsy tissues / Método aprimorado para diagnóstico da intoxicação por Nerium oleander em tecidos de necropsia

Nerium oleander is an ornamental cardiotoxic plant found in tropical and subtropical areas of the World. Its toxicity is related to the content of cardioactive glycosides, mainly oleandrin, found throughout the plant. The present study aimed to describe a new and improved method for oleandrin detection in tissue samples. The determination of oleandrin was made after extraction with a modified QuEChERS technique and measurement by UFLC-MS/MS. A total of 36 guinea pigs (Cavia porcellus) were distributed into 3 groups (n=12): control group that received only water orally (CON), and two treated groups that received hydroalcoholic oleander extract at doses of 150mg.kg-1 (OLE 150) and 300mg.kg-1 (OLE 300) in single oral dose. After three hours, fragments of heart, kidneys, liver and brain were collected for determination of oleandrin levels. The extraction and chromatographic procedures were effective for oleandrin detection and quantification in tissues, with retention time of 1.2 min and detection limit of 0.001μg g-1. The chromatographic analysis of treated guinea pigs indicated that oleandrin is distributed equally among the analyzed tissues. The developed methodology is a reliable, effective and rapid form of diagnosis of N. oleander poisoning based on necropsy tissue samples.(AU)

Nerium oleander é uma planta cardiotóxica ornamental encontrada em áreas tropicais e subtropicais do mundo. Sua toxicidade é relacionada á presença de glicosídeos cardioativos, principalmente a oleandrina, encontrada em toda a planta. O presente estudo objetiva descrever um novo e aprimorado método para detecção da oleandrina em amostras de tecido. A determinação da oleandrina foi feita após extração utilizando técnica modificada de QuEChERS e mensuração por UFLC-MS/MS. Um total de 36 cobaios (Cavia porcellus) foi distribuído em três grupos (n=12): grupo controle que recebeu apenas água por via oral (CON), e dois grupos tratados que receberam extrato hidroalcóolico de oleander nas doses de 150mg.kg-1 (OLE 150) e 300mg.kg-1 (OLE 300) em uma única dose oral. Após três horas, fragmentos do coração, rins, fígado e cérebro foram coletados para determinação dos níveis de oleandrina. A extração e procedimentos cromatográficos foram eficientes na detecção e quantificação da oleandrina nos tecidos, com tempo de retenção de 1,2min e limite de detecção de 0,001μg g-1. A análise cromatográfica dos animais tratados indicou que a oleandrina é distribuída de forma equalizada pelos tecidos analisados. A metodologia desenvolvida representa uma forma de diagnóstica segura, efetiva e rápida da intoxicação por N. oleander a partir de amostras de tecidos de necropsia.(AU)

Somatic embryogenesis, pigment accumulation, and synthetic seed production in Digitalis davisiana Heywood.

Digitalis davisiana, commonly called Alanya foxglove, from Turkey, is an important medicinal herb as the main source of cardiac glycosides, cardenolides, anthraquinones, etc. It is also known in the Indian Medicine for treatment of wounds and burns. It has ornamental value as well. Overexploitation of D. davisiana has led this species to be declared protected, and thereby encouraged various methods for its propagation. In this study, an optimized and efficient plant tissue culture protocol was established using cotyledonary leaf, hypocotyl and root explants of D. davisiana. Callus tissues were obtained from the cotyledonary leaf, hypocotyl and root segments cultured on Murashige and Skoog’s (MS) medium containing different plant growth regulators. The maximum number of somatic embryos were achieved by the MS medium containing 6-benzyladenine (1.0 mg/L BAP) or 2,4-dichlorophenoxy acetic acids (0.1 mg/L 2,4-D), which produced an average of 8.3 ± 1.5 or 5.3 ± 1.5 embryos per cotyledonary leaf, respectively. After 3 wk of culture in MS medium supplemented with 1.0 mg/L 2,4-D, callus showed a clear accumulation of orange pigmentation. Shoot regeneration was remarkably higher (14.3 indirect shoots) in a combination of α-naphthalene acetic acid (0.25 mg/L NAA) plus 3.0 mg/L BAP than 2.0 mg/L zeatin (10.3 ± 0.5 direct shoots) alone. The shoots were successfully rooted on MS medium supplemented with NAA (0.1-1.0 mg/L). In addition, synthetic seeds were produced by encapsulating shoot tips in 4% sodium alginate solution. Maximum conversion frequency of 76.6% was noted from encapsulated shoot tips cultured on 0.25 mg/L NAA with 1.0 mg/L BAP. The encapsulated shoot tips could be stored up to 60 days at 4°C. Regenerated plantlets of D. davisiana were successfully acclimatized and transferred to soil. This study has demonstrated successful preservation of elite genotypes of D. davisiana.

Concentrações de oleandrina nas folhas de Nerium oleander de diferentes cores da floração / Oleandrin concentrations in leaves of Nerium oleander of different colors of flowering

Nerium oleander é uma planta ornamental responsável por intoxicações em animais e humanos. Todas as partes da planta contém glicosídeos cardiotóxicos, principalmente a oleandrina. Alguns autores apontam que a toxicidade da planta apresentaria variação em função da cor da flor. O objetivo deste trabalho foi determinar se existe variação na concentração de oleandrina nas folhas de N. oleander de exemplares da planta com diferentes cores de inflorescências. Foram coletadas 10 amostras de folhas para cada tipo de flor (branca, rosea e vermelha). Os níveis de oleandrina foram determinados por meio de HPLC-UV após extração com metanol, precipitação com acetato de chumbo e separação em cartucho de C18. A média da concentração obtida em todas as folhas analisadas foi de 4,89mg g-1, sendo 6,20±4,08mg g-1 na variedade de flores brancas, 4,16±3,44mg g-1 na de flores roseas e 4,31±1,99mg g-1 na de flores vermelhas. As concentrações de oleandrina não apresentaram diferença estatisticamente significante entre as variedades de flores.

Nerium oleander is an ornamental plant responsible for poisoning in animals and humans. All parts of the plant contain cardiotoxic glycosides, mainly oleandrin. Some authors suggest that the toxicity of the plant would present a dependence on flower color. The objective of this study was to determine whether there is variation in the concentration of oleandrin in the leaves of N. oleander specimens of different colors of flowers. Ten samples of leaves from each variety of flower (white, pink and red) were collected. Oleandrin levels were determined by HPLC-UV after extraction with methanol, precipitation with lead acetate and separation with C18 cartridge. The average concentration obtained in all analyzed leaves was 4.89mg g-1, being 6.20±4.08mg g-1 in variety of white flowers, 4.16±3.44mg g-1 in pink flowers, and 4.31±1.99mg g-1 in red flowers. The concentrations of oleandrin did not show statistically significant difference between the varieties of flowers.

Antiproliferative cardenolides from Pentopetia androsaemifolia Decne. from the Madagascar rain forest.

Plant natural products have historically been very important to drug discovery and development, particularly in the anticancer field. This is illustrated by a discussion of the structures and activities of camptothecin and its analogues, paclitaxel (Taxol®), the vinca alkaloids vinblastine and vincristine, and podophyllotoxin and its analogues. A description of the isolation of one new and three known cardenolides from the Madagascar plant Pentopetia androsaemifolia is then provided as an example of this approach to drug discovery. The paper concludes with a brief discussion of betulinic acid, an old compound which is being developed into an anticancer and anti-HIV agent, and ipomoeassin F, an interesting antiproliferative compound isolated from a plant collected in Suriname.

Cardiac glycosides isolated from the Indian-snuff, Maquira sclerophylla Ducke

The hydroalcoholic extract of the powdered bark of the Indian-snuff Maquira sclerophylla Ducke was purified by column chromatography in silica-gel and the major cardenolide isolated from preparative TLC was identified by 1H-NMR, 1 2 C-NMR and IR analyses. The spectra showed that the active substance has strophanthidin as aglicone.