ß-sitosterol isolated from the leaves of Trema orientalis (Cannabaceae) promotes viability and proliferation of BF-2 cells.

Trema orientalis is a pioneer species in the cannabis family (Cannabaceae) that is widely distributed in Thai community forests and forest edges. The mature leaves are predominantly used as an anti-parasite treatment and feed for local freshwater fish, inspiring investigation of their phytochemical composition and bioactivity. The purpose of this work was to investigate the bioactive compounds in T. orientalis leaf extract and their cytotoxicity in the BF-2 fish cell line (ATCC CCL-91). Flash column chromatography was used to produce 25 mL fractions with a mixture solvent system comprised of hexane, diethyl ether, methanol, and acetone. All fractions were profiled with HPLC-DAD (mobile phase methanol:aqueous buffer, 60:40 v/v) and UV detection (wavelengths 256 and 365 nm). After drying, a yellowish powder was isolated from lipophilic leaf extract with a yield of 280 µg/g dry weight. Structure elucidation by nuclear magnetic resonance (NMR) indicated it to consist of pure ß-sitosterol. The lipophilic extract and pure compound were evaluated for cytotoxicity using BF-2 cells. MTT assays showed both leaf extract and pure compound at 1 µg/mL to increase cell viability after 24 h treatment. The respective half maximal inhibitory concentration (IC50) values of leaf extract and ß-sitosterol were 7,027.13 and 86.42 µg/ml, indicating a lack of toxicity in the BF-2 cell line. Hence, T. orientalis can serve as a source of non-toxic natural lipophilic compounds that can be useful as bioactive ingredients in supplement feed development.

Desempenho de sementes e plântulas de rabanete sob ação de extrato aquoso de folhas de Trema micrantha (Ulmaceae) / Performance of radish seeds and seedlings under action of aqueous extract of leaves of Trema micrantha (Ulmaceae)

O trabalho objetivou avaliar o efeito alelopático de extrato aquoso de folhas de Trema micrantha sobre germinação de sementes e crescimento inicial de plântulas de rabanete. Para os extratos foram utilizadas folhas maduras e frescas nas concentrações 2, 4 e 8% (p/v), com aferição do pH e potencial osmótico. No bioensaio de germinação, foram empregadas cinco repetições de 25 sementes distribuídas em placas de petri forradas com papel Germitest, umedecido com 7 mL de extrato ou água e mantidos a 25 ºC em B.O.D., por cinco dias. Foram determinados a porcentagem de germinação (PG), velocidade de germinação (VG), índice de velocidade de germinação (IVG) e índice de efeito alelopático (RI). No bioensaio de crescimento inicial, foram avaliados o comprimento da radícula e do hipocótilo, a biomassa fresca e seca, o conteúdo de água e os teores de clorofila a, b e total. Todos os atributos de germinação, PG, VG, IVG e RI, foram afetados negativamente pelos extratos de T. micrantha. O crescimento inicial foi influenciado pelos extratos, ocorrendo redução do comprimento da radícula e aumento do hipocótilo. O extrato não alterou a biomassa fresca e seca e os teores de clorofila. Extratos aquosos de folhas de Trema micrantha exerceram ação alelopática sobre a germinação e crescimento inicial do rabanete, não influenciando na inibição da síntese de pigmentos fotossintéticos.

This study aimed to evaluate the allelopathic effects of aqueous extracts of Trema micrantha leaves on seed germination and early growth of seedlings of radish (Raphanus sativus L.). Leaf extracts were prepared at concentrations of 2, 4 and 8%. pH and osmotic potential were also performed from the extracts. Germination bioassay consisted of five replicates of 25 seeds of radish distributed in Petri dishes with germitest paper and 7 mL of extract or water and kept at 25°C in B.O.D. for five days. Germination percentage (PG), germination speed (VG), germination speed index (IVG) and index of allelopathic effect (RI) were determined. For bioassay initial growth, seeds were germinated until they reach 2 mm protrusion radicle and transferred to Gerbox containing germitest paper and 15 mL of extract or water, kept seven days at 25°C in B.O.D. for evaluating length of the radicle and hypocotyl, fresh and dry mass, water content and content of chlorophyll (a, b and total). Leaf extracts from T. micrantha affected negatively all parameters examined for germination (PG, VG, IVG and RI). The extracts affected the initial growth, causing reduction of the radicle length and stimulating the growth of hypocotyls, but not interfered on fresh and dry weight and content of chlorophyll. Aqueous extracts of T. micrantha leaves exerted allelopathic action on germination and early growth of radish, but not caused inhibition of synthesis of photosynthetic pigments.

Trema micrantha como causa de pneumopatia tóxica em ovinos: reprodução experimental / Trema micrantha as cause of toxic pneumopathy in sheep: experimental reproduction

A intoxicação experimental por Trema micrantha em cinco ovinos é descrita. Quatro ovinos apresentaram sinais clínicos respiratórios acentuados e morte após ingestão da terceira dose de folhas da planta. As manifestações clínicas mais frequentes nesses casos foram taquipneia, dispneia, retração ritmada das narinas, mucosas cianóticas, corrimento nasal mucoso, hipertermia, aumento de volume abdominal bilateral e na região parotídea, incluindo crepitação à palpação, membros lateralmente afastados ao caminhar e decúbito esternal. T. micrantha mostrou-se tóxica para ovinos na dose de 20-50g/kg de peso animal. Na necropsia desses ovinos foram observados, além das mucosas cianóticas, enfisema subcutâneo em região cervical ventral ou porção mediastinal dorsal, pulmões não colabados, pesados, com impressão das costelas na superfície e conteúdo espumoso vermelho em traqueia e brônquios, além de múltiplas petéquias subpleurais. Na avaliação histológica predominaram alterações pulmonares, com espessamento de septos alveolares por proliferação difusa de pneumócitos tipo II, conferindo aspecto adenomatoso a algumas áreas. Os pneumócitos apresentavam núcleo volumoso, hipercromático, ora bizarro ou eram multinucleados, com nucléolos evidentes e, em algumas áreas, os pneumócitos estavam descamados para a luz alveolar, ora formando sincícios. Havia também proliferação do epitélio bronquiolar, com formação de mais de uma camada celular, núcleos hipercromáticos e volumosos e redução na quantidade de cílios. As alterações proliferativas dos pneumócitos e do epitélio bronquiolar foram evidenciadas pela imunomarcação anti-citoqueratina e anti-Ki-67 e, para a diferenciação entre pneumócitos e macrófagos alveolares, foi empregada imuno-histoquímica anti-mieloide/histiócitos (MAC387). Um ovino apresentou quadro clinico-patológico de insuficiência hepática aguda, com necrose hepatocelular acentuada após ingestão de 25g/kg de T. micrantha. A intoxicação experimental por ...

This paper describes the experimental Trema micrantha poisoning in sheep. Four from five sheep showed marked respiratory distress and death after ingestion of the third dose of plant leaves. The most frequent clinical findings were tachypnea, dyspnea, rhythmic contraction of the nostrils, cyanotic mucous membranes, mucous nasal discharge, hyperthermia, abdominal distension, increased volume and crepitation of parotid region, walking with the legs kept away laterally and sternal recumbency. T. micrantha proved to be toxic to sheep at 20 to 50g/kg bodyweight. At necropsy, we observed cyanotic mucous membranes, subcutaneous emphysema in the ventral area of cervical region or dorsal mediastinum, heavy and collapsed lungs with rib markings, red frothy fluid in the lumen of the trachea and primary bronchi, and several subpleural petechiae. Microscopic examination of lung tissue sections revealed that the tissue was characterized by thickening of the alveolar septa by diffuse type II pneumocytes proliferation, conferring an adenomatous appearance to some areas. These pneumocytes had large and hyperchromatic, sometimes bizarre nucleus or were multinucleate, with evident nucleoli. In some areas desquamated pneumocytes were found in the alveolar lumen, sometimes forming syncytia. There also was bronchiolar epithelium proliferation, conferring to the mucosa more than one epithelial cell layer, these cells had large and hyperchromatic nuclei and reduction in the amount of cilia. The proliferative changes of pneumocytes and bronchiolar epithelium were evaluated by anti-cytokeratin and anti-Ki-67 immunostaining. In order to differentiate pneumocytes and alveolar macrophages, we used anti-monocyte marker (MAC387). One sheep showed clinicopathological findings of acute liver failure with hepatocellular necrosis after ingestion of 25g/kg T. micrantha. The experimental poisoning by T. micrantha in sheep resulted in predominantly respiratory clinicopathological ...

Novos aspectos sobre a intoxicação por Trema micrantha (Cannabaceae) em equídeos / New aspects of Trema micrantha (Cannabaceae) poisoning in equids

Trema micrantha é uma planta arbórea distribuída amplamente no Brasil. Descrevem-se nesse trabalho novos aspectos epidemiológicos e patológicos relacionados com a intoxicação por essa planta em equídeos. Dois equinos adultos da raça Crioula e dois asininos, de localidades distintas, foram intoxicados naturalmente por Trema micrantha, após consumirem grande quantidade da planta que ficou disponível a eles após uma poda. Além disso, um cavalo adulto, de outra propriedade, que estava passando por restrição alimentar, se intoxicou após consumir as partes baixas de um grande número de árvores jovens de T. micrantha. Clinicamente, em todos os equídeos a doença se caracterizou por alterações neurológicas, que apareceram três a quatro dias após o consumo da planta, com curso clínico de dois a quatro dias (abreviadas por eutanásia). Os principais achados de necropsia foram observados no fígado, que estavam levemente aumentados e com padrão lobular evidente e no sistema nervoso central (SNC), com múltiplas áreas amareladas, focos de malacia e hemorragia, principalmente, no tronco cerebral e cerebelo. Histologicamente, havia necrose hepática aguda, edema acentuado no SNC com degeneração fibrinoide da parede de vasos associada a hemorragia e trombose e, frequentemente, com infiltrado de neutrófilos. Outras alterações observadas nos encéfalos foram: grande quantidade de astrócitos de Alzheimer tipo II na substância cinzenta, acúmulos de células Gitter e degeneração Walleriana próxima a focos de lesões vasculares.

Trema micrantha is a tree widely distributed in Brazil. This paper describes new epidemiological and pathological aspects of this plant poisoning in equids. Two adult horses, Criollo breed and two donkeys from different locations were naturally poisoned by Trema micrantha after consuming large amounts of the plant which was available to them due to pruning. Also an adult horse from another ranch, which was undergoing dietary restriction, was poisoned after consuming the lower parts of a large number of young T. micrantha trees. Clinically, in all cases, the disease was characterized by neurological symptoms that started 3 to 4 days after consumption of the plant, with a clinical course that varied 2 to 4 days (abbreviated by euthanasia). The main gross findings were in the liver, which had slightly increased lobular pattern evident, and in the central nervous system (CNS) with multiple yellowish areas, foci of malacia and hemorrhage, especially in the brainstem and cerebellum. Histologically, there was acute liver necrosis with marked edema of the CNS with fibrinoid vessel wall degeneration, hemorrhage and thrombosis as well as neutrophil infiltration. Other changes in the brain were Alzheimer type II astrocytes in the gray matter, accumulation of Gitter cells and Wallerian degeneration near to vascular lesion foci.

Differences on post-fire regeneration of the pioneer trees Cecropia glazioui and Trema micrantha in a lowland Atlantic Forest

A study of natural post-fire succession was carried out in a disturbed vegetation around fragments of the Atlantic Rain Forest (National Biological Reserve of Poço das Antas (22° 30’- 22° 33’S, 42° 15’- 42° 19’ W), Rio de Janeiro State). All the pre-fire individuals of Cecropia and Trema in the area were numbered with plastic labels.In order to check for the presence of new sprouts and mortality, two other censuses were carried out, at 3 and 12 months after the fire.The dominant species were:Pteridium aquilinum,Panicum maximum,Trema micrantha and Cecropia glazioui. Few days after the passage of fire, grasses and ferns spread their area,while the stands of Trema and Cecropia were completely burned. Most of individuals of Cecropia produced some sprouts while most of individuals of Trema died.However,a great number of seedlings of Trema were recruited while only one single seedling of Cecropia were observed during a period of one year.Most of these seedlings died through the year while the sprouts were already reproducing.The uses of Cecropia in places where fire is recurrent could be more appropriate because of its higher chance of survival and faster recovering ability after fire

Se realizó un estudio sobre la sucesión natural después del fuego en una vegetación alrededor de fragmentos de la selva lluviosa atlántica (Reserva Biológica Nacional de Poço das Antas (22° 30’-22° 33’ S, 42° 15’- 42° 19’ W), Estado de Río de Janeiro).Todos los individuos de Cecropia y Trema previos del fuego en el área fueron numerados con marcas plásticas. Se realizaron otros dos censos, 3 y 12 meses después del fuego con el objetivo de detectar mortalidad y buscar la presencia de nuevos brotes. Las especies dominantes fueron: Pteridium aquilinum, Panicum maximum, Trema micrantha y Cecropia glazioui. Pocos días después del paso del fuego,los pastos y helechos se dispersaron por el área, mientras que los troncos de Trema y Cecropia estaban completamente quemados. La mayoría de los individuos de Cecropia produjeron algunos rebrotes mientras que la mayoría de los individuos de Trema murieron. Sin embargo, un gran número de plántulas de Trema fueron reclutadas mientras que únicamente una plántula de Cecropia fue observada durante el período de un año. La mayoría de estos plántulas murieron a lo largo del año, mientras que los brotes se estan ya reproduciendo. Los usos de Cecropia en lugares donde el fuego es recurrente puede ser más apropiado debido a su mayor chance de sobrevivir y a su habilidad de recuperarse más rápido después del fuego