Shiitake (Lentinula edodes (Berkeley) Pegler) extracts as a modulator of micronuclei induced in HEp-2 cells

Shiitake (Lentinula edodes (Berkeley) Pegler) is one of the most consumed mushrooms, for both therapeutic purposes and as food, therefore, the study of its biological properties is of great interest for producers and consumers. Aqueous extracts of the shiitake mushroom (L. edodes (Berkeley) Pegler) were evaluated by the micronucleus test (MN) in HEp-2 cells in vitro, to analyze their possible mutagenic and antimutagenic activities. None of the three extract concentrations tested (0.5, 1.0 and 1.5 mg/mL) presented mutagenicity at any of the preparation temperatures (4 degrees C, 22 +/- 2 degrees C and 60 degrees C). In the antimutagenicity evaluation, all extract concentrations at all preparation temperatures presented a strong protective activity for the HEp-2 cells in response to the alkylating agent methyl methanesulfonate (MMS) in the different treatment protocols: pre-treatment, simultaneous treatment and post-treatment. The extracts prepared at 22 +/- 2 degrees C presented the lowest frequencies of MN in the evaluations of mutagenicity and antimutagenicity, indicating these as the best option for potential therapeutic use. (c) 2006 Elsevier Ltd. All rights reserved.

Cytotoxicity, genotoxicity and antimutagenicity of hexane extracts of Agaricus blazei determined in vitro by the comet assay and CHO/HGPRT gene mutation assay

Agaricus blazei Murrill ss. Heinem, known as the sun mushroom or himematsutake, is a basidiomycete native to Brazil, which is popular for its medicinal properties. The aim of this study was to test hexane extracts (one fraction and its four sub-fractions) of A. blazei for bioactivity in cultured mammalian cells (CHO-K1). The comet assay, the colony forming assay (CFA) and CHO/HGPRT gene mutation assay were used respectively to determine genotoxicity, cytotoxicity and antimutagenicity of these extracts at different concentrations. The cells were incubated in culture medium and treated for 3 h according to the standard protocol for each assay. The DNA damage-inducing agent ethylmethane sulfonate (EMS) was utilized as the positive control and also in combination with extracts to test for a protective effect. Statistical analysis of the data was performed using analysis of variance (ANOVA) and Tukey's test. A relationship between cytotoxicity and genotoxicity could be established and two extracts EH6B and EH6D showed a protective tendency, while the others did not, with the primary extract EH6 causing the most substantial damage to genetic material. These findings warrant more in-depth studies of the active principles of this mushroom. (c) 2005 Elsevier Ltd. All rights reserved.

Mushroom-derived preparations in the prevention of H2O2-induced oxidative damage to cellular DNA

Aqueous extracts of the sporophores of eight mushroom species were assessed for their ability to prevent H2O2-induced oxidative damage to cellular DNA using the single-cell gel electrophoresis (Comet) assay. The highest genoprotective effects were obtained with cold (20°C) and hot (100°C) water extracts of Agaricus bisporus and Ganoderma lucidum fruit bodies, respectively. No protective effects were observed with Mushroom Derived Preparations (MDPs) from Flammulina velutipes, Auricularia auricula, Hypsizygus marmoreus, Lentinula edodes, Pleurotus sajor-caju, and Volvariella volvacea. These findings indicate that some edible mushrooms represent a valuable source of biologically active compounds with potential for protecting cellular DNA from oxidative damage. © 2002 Wiley-Liss, Inc.

Mycelial growth of two Lentinula edodes strains in culture media prepared with sawdust extracts from seven eucalyptus species and three eucalyptus clones

The in vitro mycelial growth of Lentinula edodes strains LE-95/01 and LE-96/18 were evaluated in solid culture media prepared with sawdust extracts from seven eucalyptus species (E. saligna, E. grandis, E. urophylla, E. pellita, E. paniculata, E. citriodora, and E. camaldulensis) and three eucalyptus clones (E. grandis × E. urophylla hybrids). Evaluations were made every 48 hours by means of colony diameter measurements (mean of four transversely-oriented measurements), during ten days of incubation in the dark at 25ºC ±1°C. The experimental design consisted of randomized blocks, and treatment means were compared by Tukey test. The culture medium prepared from E. citriodora sawdust extract was the most promising to grow L. edodes strains LE-96/18 and LE-95/01. L. edodes strain LE-96/18 presented the fastest mycelial growth after incubation for ten days, regardless of sawdust extract type used in the culture medium.

Influence of Lentinus edodes and Agaricus blazei extracts on the prevention of oxidation and retention of tocopherols in soybean oil in an accelerated storage test

This study aimed to evaluate the influence of the methanol extracts of mushrooms Lentinus edodes and Agaricus blazei on the retention of tocopherols in soybean oil, when subjected to an accelerated storage test. The following treatments were subjected to an accelerated storage test in an oven at 60 A degrees C for 15 days: Control (soybean oil without antioxidants), TBHQ (soybean oil + 100 mg/kg of TBHQ), BHT (soybean oil + 100 mg/kg of BHT), L. edodes (soybean oil + 3,500 mg/kg of L. edodes extract) and A. blazei (soybean oil + 3,500 mg/kg of A. blazei extract). The samples were analyzed for tocopherols naturally present in soybean oil and mass gain. The results showed, the time required to reach a 0.5% increase in mass was 13 days for TBHQ and 15 days for A. blazei. The content of tocopherols for TBHQ was 457.50 mg/kg and the A. blazei, 477.20 mg/kg.

Activity of sesame leaf extracts to the leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera : Formicidae)

Crude extracts of new and old sesame leaves (Sesamum indicum, Linaeus) obtained by a sequential process of extraction were tested through contact experiments to investigate their toxicity to Atta sexdens rubropilosa (Forel) workers. Methanol extract of old leaves was toxic to leaf-cutting ants at the lower tested concentration (20mg.ml(-1)). This extract was fractioned and their fractions were tested showing a toxic effect of the methanol fraction. The methanol fraction was fractioned again and the sub fraction responsible for worker's toxicity is composed of some sugars.

Activity of sesame leaf extracts against the symbiotic fungus of Atta sexdens L.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Biological control of Bradysia matogrossensis (Diptera : Sciaridae) in mushroom cultivation with predatory mites

Larvae of Sciaridae flies (Diptera) cause considerable damage to the mushroom Agaricus blazei (Murrill) ss. Heinemann in Brazil. Brazilian growers have had considerable difficulties in controlling this pest. The objective of this work was to test the effect of the predatory Laelapidae mite Stratiolaelaps scimitus (Womersley) as a control agent of Bradysia matogrossensis (Lane) in cultures of A. blazei. The work corresponded to an evaluation of the efficiency of that predator when released in boxes containing each about 15 L of commercial mushroom compost naturally infested with the pest. The results showed a significant effect of that predator on the population of B. matogrossensis. The release of either 665 or 1330 S. scimitus per box significantly reduced the pest population to levels that, according to grower's experience, apparently could not cause considerable damage. The positive results obtained warrant the conduction of complementary studies to determine the lowest rates of the predator that could still produce acceptable levels of control.

The predatory mite Stratiolaelaps scimitus as a control agent of the fungus gnat Bradysia matogrossensis in commercial production of the mushroom Agaricus bisporus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ticks' response to feeding on host immunized with glandular extracts of Rhipicephalus sanguineus females fed for 2, 4, and 6 days. I. Inactivity or early degeneration of salivary glands?

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In vitro acaricidal activity of neem (Azadirachta indica) seed extracts with known azadirachtin concentrations against Rhipicephalus microplus

The effect of four extracts from neem seeds (Azadirachta indica) containing 2000, 5000, 9000 and 10,000 ppm of azadirachtin A (AZA), quantified by high-performance liquid chromatography (HPLC) and diluted to 1.25%; 2.5%; 5.0%; 10.0% and 12.8% was verified by in vitro tests with engorged females and larvae of the cattle tick Rhipicephalus micro plus. The results from the bioassays with the engorged females showed that the main toxic effect of the extracts was reduction of the reproductive parameters, with a sharp drop in the number of eggs laid and the hatching rate, mainly when the extracts were diluted to 10.0% and 12.8%. The product effectiveness (PE) calculations for all the solutions tested showed that the AZA solution at 10,000 ppm (N10) was the most effective. However, statistical analysis of the PE data obtained for the proportional AZA concentrations in the different diluted extracts showed significance (P<0.05) of the effects included in the model (extract dilution, principle effect (classificatory) of the assay (extract) and the interaction between the two), indicating significant variations due to the dilution, the test and the interaction between the two factors in the tests with engorged females. For solutions N2, N5, and N9, it was not possible to estimate LC(90) values in the dilution range tested. The lowest LC(50) was observed for extract N5, and although extract N10 was the only extract for which the LC(90) could be estimated within the range tested, the LC(50) was higher than for N5 and N9. These results suggest that substances other than AZA present in the extracts influenced the efficacy, especially up to a certain LC range. In the tests with larvae, no mortality was observed, indicating zero effectiveness of all the extracts tested. The results of the tests with engorged females showed that the neem extracts had acaricide activity, inhibiting egg laying and the larval hatching rate. Complementary studies are necessary to develop new methods to isolate and/or identify other substances besides AZA contained in this plant, to enable using products made from it as acaricides. (C) 2011 Elsevier B.V. All rights reserved.

Antimutagenic effect of Lentinula edodes (BERK.) Pegler mushroom and possible variation among lineages

This study was performed to evaluate the efficiency of four different lineages (95/01, L1, 96/22 and JABK) of Lentinula edodes (BERK.) Pegler mushroom (shiitake) for inhibiting the N-ethyl-N-nitrosourea (ENU) clastogenicity in vivo. Male Swiss mice (10 animals/group) were treated during 15 consecutive days with dried mushroom added to basal diet under three different concentrations (1, 5 and 10%). At day 15, mice were intraperitoneally injected with ENU (50 mg/kg body weight) and sacrificed 24 h later for evaluation of micronucleated bone marrow polychromatic erythrocytes (MNPCE). Negative and positive controls (10 animals each), receiving basal diet and saline or ENU ip injection, respectively, were also evaluated. Results showed that pretreatments with diets containing the lineages 95/01, L1 and 96/22 reduce the frequencies of MNPCE induced by ENU. The absence of an antimutagenic activity for the lineage JABK might be related to intrinsic differences among the lineages such as biochemical composition. Taken together, our data show that the differences in protective activities of the mushrooms need to be clarified in further studies and the mechanisms for such activities need to be investigated. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Interaction between resistant tomato genotypes and plant extracts on Bemisia tabaci (Genn.) biotype B

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

APPLICATION of THE HAZARD ANALYSIS and CRITICAL CONTROL POINT (HACCP) SYSTEM on THE MUSHROOM PROCESSING LINE FOR FRESH CONSUMPTION

The Hazard Analysis and Critical Control Point (HACCP) is a preventive system that intends to guarantee the safety and harmlessness of food. It improves the quality of products as it eliminates possible defects during the process, and saves costs by practically eliminating final product inspection. This work describes the typical hazards encountered on the mushroom processing line for fresh consumption. Throughout the process, only the reception stage of mushrooms has been considered a critical control point (CCP). The main hazards at this stage were: the presence of unauthorised phytosanitary products; larger doses of such products than those permitted; the presence of pathogenic bacteria or thermo-stable enterotoxins. Putting into practice such knowledge would provide any industry that processes mushrooms for fresh consumption with a self-control HACCP-based system for its own productions.

Growth of the Symbiotic Fungus of the Grass-cutting Ant Atta capiguara (Hymenoptera: Formicidae): Effect of Grass Extracts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)