ABSTRACT
Basidiocarp of Agaricus blazei (=Agaricus brasiliensis; =Agaricus subrufescens) is used as teas or capsules due to its antineoplastic effect but there are few reports of using mycelium for this purpose. The objective of this study was to evaluate the antineoplastic activity on sarcoma 180 cells implanted in mice of two forms of preparation of the mycelium from two A. blazei strains grown in culture medium with different concentrations of isolated soy protein. Mycelia were grown in Pontecorvo medium with different concentrations of isolated soybean protein (ISP). Mycelial hot water extract, moistened mycelial powder, hot water extract of green tea, Ifosfamida(®) (ifosfamide drug), and saline solution were administered daily by gavage in mice with sarcoma 180 cells to evaluate antineoplastic activity. It was concluded that antineoplastic activity was the same for both strains, except when used as moistened mycelial powder, which rules out the use of mycelial powder in capsules. Mycelial hot water extract had high antineoplastic activity with lower metabolic demand on the spleen and maintenance of normal blood parameters. Mycelial growth in different ISP concentrations had the same antineoplastic activity. Also the vegetative mycelium was as effective as the basidiocarp for sarcoma 180 tumor inhibition. Green tea was as effective as mycelial hot water extract.
Subject(s)
Agaricus/chemistry , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/therapeutic use , Mycelium/chemistry , Plant Extracts/administration & dosage , Plant Extracts/therapeutic use , Sarcoma 180/drug therapy , Animals , Culture Media/chemistry , Drug Administration Routes , Drug Compounding , Female , Ifosfamide/therapeutic use , Mice , Phytotherapy , Sarcoma 180/pathology , Soybean Proteins/pharmacologyABSTRACT
Alternative substrates for cryopreservation at -20 °C have been little explored for basidiomycetes and could bring new possibilities of lower cost cryopreservation. Nevertheless, freezing temperatures between -15 and -60 °C are very challenging because they frequently result in cryoinjuries. The objective of this study was to evaluate substrates associated to cryoprotective agents for Pleurotus ostreatus cryopreservation at -20 or -70 °C in order to develop alternative techniques for basidiomycete cryopreservation. P. ostreatus was grown on potato dextrose agar or whole grains of oat, wheat, rice or millet and transferred to cryovials with cryoprotective solution with 1 % dimethyl sulfoxide, 5 % glycerol, 10 % saccharose, 4 % glucose, 6 % polyethylene glycol-6000 or 5 % malt extract. The mycelium in the cryovials were cryopreserved at -20 or -70 °C and recovered for evaluation of the mycelial growth viability after 1 and 3 years. Both substrates and cryoprotectants affect the viability of the mycelial growth cryopreserved at -20 or -70 °C; wheat grains combined with cryoprotectants such as saccharose or glucose are effective for keeping mycelium viable after cryopreservation at -20 °C for 1 or 3 years; for cryopreservation at -70 °C after 1 or 3 years, any substrate combined with any cryoprotectant is effective for preserving the mycelium viable, except for millet grains with polyethylene glycol after 3 years; semi-permeable cryoprotective agents such as saccharose and glucose are the most effective for cryopreservation at -20 or -70 °C for at least 3 years.
ABSTRACT
O extrato alcoólico dos frutos da espécie nativa Allophylus edulis (A.St.-Hil., Cambess. & A. Juss.) Radlk. (Sapindaceae), popularmente conhecida como ?vacum?, ?fruto-de-pombo?, ?vacunzeiro?, ?chal-chal?, ?murta vermelha?, foi avaliado no presente trabalho quanto às atividades citotóxicas, antioxidantes e anticolinesterásicas. O extrato dos frutos mostrou baixa toxicidade pelo bioensaio de letalidade frente à náuplios de Artemia salina Leach (CL50 > 1000 µg mL-1), não sendo considerado citotóxico. As propriedades antioxidantes foram investigadas in vitro pelo método de DPPH e foram consideradas bastante significativas (CI50 = 46,4 µg mL-1). Apenas um componente do extrato mostrou atividade anticolinesterásica pelo ensaio enzimático de inibição de acetilcolinesterase. Estes resultados sugerem que os frutos de Allophylus edulis apresentem baixa toxicidade e sejam considerados uma fonte de metabólitos bioativos, podendo ser indicados em terapias de doenças relacionadas com a presença de radicais livres. Tanto a atividade antioxidante quanto a atividade anticolinesterásica estão sendo relatadas pela primeira vez para esta espécie.
The alcoholic fruit extract of native species Allophylus edulis (A.St.-Hil., Cambess. & A. Juss.) Radlk. (Sapindaceae), commonly known as ?vacum?, ?fruto-de-pombo?, ?vacunzeiro?, ?chal-chal?, ?murta vermelha?, was evaluated in this study regarding its cytotoxic, antioxidant and anticholinesterase activities. The fruit extract showed low toxicity to bioassay lethality for Artemia salina Leach (LC50> 1000 µg mL-1) and not considered cytotoxic. The antioxidant properties were investigated in vitro by DPPH method and were considered highly significant (IC50 = 46.4 µg mL-1). Only one component of this extract showed anticholinesterase activity by enzymatic assay of inhibition of acetylcholinesterase. These results suggest that Allophylus edulis fruits have low toxicity and are considered a source of bioactive metabolites, may be prescribed in the therapy of diseases related to the presence of free radicals. Both the antioxidant and anticholinesterase activities for this species have been reported for the first in this study.
Subject(s)
Sapindaceae , Cytotoxins , AntioxidantsABSTRACT
A técnica de transferências periódicas de fragmentos de micélio para novo meio de cultura é a mais utilizada para a preservação de Agaricus blazei. Entretanto, esta técnica apresenta maior risco de contaminação, degeneração genética e perda de caracteristicas biológicas. O desenvolvimento de técnicas de preservação que permitam a manutenção da viabilidade da espécie por mais tempo e a um menor custo é de interesse biotecnológico. Desse modo, o objetivo deste trabalho foi avaliar a viabilidade de A. blazei crescido em dois meios de cultivo e preservado à +4 ºC ou -20 ºC em diferentes recipientes de contenção. O fungo foi crescido em meio de ágar-extrato de malte ou ágar-grão de trigo moído e preservado à +4 ºC ou -20 ºC em diferentes recipientes de contenção, simples ou duplos, com adição de soluções aquosas de glicerol, sacarose, glicose, água ultrapura ou sem adição de crioprotetor. Após 1 ou 12 meses o micélio preservado foi transferido para ágar-extrato de malte para avaliação da viabilidade micelial. Os crioprotetores glicerol, sacarose e glicose, associados com o meio de cultura ágar-extrato de malte ou ágar-grão de trigo moído, em recipiente de contenção simples ou duplo são efetivos para preservação à +4 ºC por períodos curtos, um mês, mas não são efetivos para períodos longos, 12 meses. Os crioprotetores, meios de cultivo e recipientes de contenção simples ou duplos não são efetivos para criopreservação do fungo à -20 ºC. Os recipientes simples são tão eficientes quanto os recipientes duplos para evitar contaminações e preservar o fungo.
Continuous mycelial subculturing is frequently used for the preservation of Agaricus blazei. However, this technique has a higher risk of contamination, genetic degeneration and loss of biological characteristics. The development of preservation techniques that allow maintaining the viability of this species longer and at lower costs is of biotechnological interest. Thus, the objective of this study was to evaluate the viability of A. blazei grown in two culture media and preserved at +4 ºC or -20 ºC in different containment vessels. The fungus was grown on malt extract agar or grounded wheat grain agar culture medium and preserved at +4 ºC or -20 ºC in different containment vessels, single or double ones, with the addition of aqueous solutions of glycerol, saccharose, glucose, ultrapure water or without addition of cryoprotectant. After 1 or 12 months, the preserved mycelium was transferred to malt extract agar for assessment of mycelial viability. Glycerol, saccharose and glucose associated with malt extract agar or grounded wheat grain agar culture medium, in single or double containment vessels, are effective for preservation at +4 ºC for a short period, one month, but they are not effective for a longer period, 12 months. Cryoprotectants, culture media and single or double containment vessels are not effective for fungus cryopreservation at -20 ºC. Simple containment vessels are as efficient as double ones to prevent contamination and to preserve the fungus.
La técnica de transferencias periódicas de fragmentos de micelio para nuevo medio de cultura es la más utilizada para la preservación de Agaricus blazei. Sin embargo, esta técnica presenta mayor riesgo de contaminación, degeneraciones genéticas y pérdidas de características biológicas. El desarrollo de técnicas de preservación que permitan la manutención y viabilidad de la especie por más tiempo y con un costo más bajo es de interés biotecnológico. De esta manera, el objetivo de este estudio fue evaluar la viabilidad de A. blazei sembrado en dos medios de cultivo y preservados en +4 ºC o -20 ºC en diferentes recipientes de contención. El hongo se cultivó en medio de extracto de agar de malta o agar de grano de trigo molido y preservado en +4 ºC o -20 ºC en diferentes recipientes de contención simple o doble, con adición de soluciones acuosas de glicerol, sacarosa, glucosa, agua ultra pura o sin adición de crioprotector. Después de 1 o 12 meses, el micelio preservado fue transferido para extracto de agar de malta para evaluación de la viabilidad del micelio. Los crioprotectores glicerol, sacarosa y glucosa, asociados con el medio de cultura extracto de agar de malta o de agar de grano de trigo molido, en recipiente de contención simple o doble son eficaces para la preservación a +4 ºC por períodos cortos, un mes, pero no son eficaces por períodos largos, como 12 meses. Los crioprotectores medios de cultivo y recipientes de contención simples o dobles no son eficaces para la criopreservación del hongo a -20 ºC. Recipientes simples son tan eficaces como los dobles para evitar contaminaciones y preservar el hongo.
Subject(s)
Agaricus/growth & development , Cryoprotective Agents , Culture Media , Mycelium/isolation & purification , Glycerol/administration & dosage , Glucose/administration & dosage , Sucrose/administration & dosage , Agar/administration & dosageABSTRACT
Different maturation phases of basidiocarp could affect the bioactivity and concentration of some active substances. A. brasiliensis Wasser et al.(A. blazei Murrill) has shown antitumor activity that could be related to the antioxidant activity. However there is no information of the best basidiocarp maturation phase for extracting antioxidant substances in order to determine the moment of harvesting in mushroom cultivation. The objective of this work was to evaluate the antioxidant activity of A. brasiliensis strains on different basidiocarp maturation phases. The best condition for extraction of A. brasiliensis antioxidants is with methanol as solvent at 60 ºC for 60 min. Strains with closed basidiocarp have higher antioxidant activity than with opened basidiocarp. Antioxidant activity varies in each strain. It was concluded that A. brasiliensis is a natural source of antioxidant compounds. Also there is higher antioxidant activity in closed than opened caps and consequently higher functional activity. It reinforces the synergic action among different A. brasiliensis compounds as a functional food and the importance of further investigation for isolation and characterization of antioxidant substances of A. brasiliensis. It also determines the best harvest period in order to obtain the highest antioxidant activity from basidiocarp.
