Biodegradation of agave Comiteco bagasse by Pleurotus spp.: a source of cellulases useful in hydrolytic treatment to produce reducing sugars.

This study aimed to determine the production parameters of five strains of Pleurotus spp. during their cultivation on agave Comiteco bagasse, as well as the feasibility of using cellulolytic extracts to produce reducing sugars in the same bagasse. After cultivation, the basidiome production parameters varied between 41.2 and 65.7% (biological efficiency), 0.17 and 0.30 (yield), 0.60 and 0.90% (production rate), 16.4 and 41.1% (Bioconversion) and 9.4 and 21.3 g (mean mushroom weight). At day 15 of growth, P. djamor showed the highest ß-glucosidase activity (43.95 ± 4.5 IU/g); on day 33. The same strain had the highest endoglucanase activity (21.12 ± 0.5 IU/ml). Both extracts were partially purified, and the kinetic parameters Vmax and Km were estimated (20.83 µmole/ml sec and 232.01 µmole/ml for ß-glucosidase and 685.01 µmole/ml sec and 1,240.34 µmole/ml for endoglucanase). In the enzymatic hydrolysis assay, the highest concentration of reducing sugars (43.13 ± 1.09 g/L; 0.21 g/g bagasse) was obtained by a mixture of the two partially purified extracts acting synergistically after 48 h and with a pH adjustment. The results suggest that the use of agave Comiteco bagasse for cultivating edible mushrooms while obtaining cellulolytic extracts is an alternative treatment for waste reduction and valorization of agro-industrial by-products.

Nematocidal activity of hydroalcoholic extracts of spent substrate of Pleurotus djamor on L3 larvae of Haemonchus contortus.

The objective of this study was to evaluate and compare the in vitro lethal effect of the hydroalcoholic extract of the spent substrate of Pleurotus djamor ECS-123, obtained at 15 days of colonization (SPS) and at the first (SPS1) and second (SPS2) harvests, against infective larvae L3 of Haemonchus contortus. The in vitro lethal effect was evaluated by the L3 larval mortality test (LM) using six concentrations: 1.25, 2.5, 5, 10, 20, and 40 mg/mL, with ivermectin and thiabendazole (5 mg/mL) as controls. The first harvest extract (SPS1) of strain ECS-123 was subjected to liquid-liquid bipartition, which resulted in two fractions: aqueous (PdAcO) and ethyl acetate (PdAct). The chemical fractionation of PdAct with the highest mortality rate (80.11 %) was carried out with open-column chromatography, giving a total of 13 fractions, which were analyzed by thin-layer chromatography (TLC) and grouped into 5 mixtures (R1;1-3, R2;4-7, R3;8-9, R4;10-11 and R5;12-13). Subsequently, the mixtures were evaluated against H. contortus L3 larvae. Finally, the components of the mixtures with the highest nematocidal effects were evaluated by gas chromatography coupled to mass spectrometry (GC-MS). The data were analyzed with a completely randomized design through ANOVA using the generalized linear model (GLM) with the "R" program. The purification and characterization of R4 and R5 by GC-MS revealed the presence of the following compounds: veratryl alcohol, 4-hydroxy-3,5,5 trimethyl-4-[3-oxo-1-butenyl]-2- cyclohexen-1-one, caffeine and 5,6-dimethoxy-1(3 H) isobenzofuranone. This information allowed for the identification of nematocidal compounds in the degraded substrate of P. djamor, an activity that had not been reported previously.

Chemical Composition of an Anthelmintic Fraction of Pleurotus eryngii against Eggs and Infective Larvae (L3) of Haemonchus contortus.

This study was aimed at evaluating the in vitro effect of the edible mushroom (EM) Pleurotus eryngii against the eggs and larvae (L3) of Haemonchus contortus. The evaluation included acetone (AE) and hydroalcoholic (HA) extracts of the following strains: ECS-1138, ECS-1156, ECS-1255, ECS-1258, ECS-1261, ECS-1282, and ECS-1292. The HA extract of the ECS-1255 strain showed the highest effect on mortality rates of L3 (18.83%) at 20 µg/mL. After subjecting this HA extract to a normal phase chromatography column, five fractions were obtained; fraction F5 (100% MeOH) was the most effective against eggs, with hatching inhibition percentages of 88.77 and 91.87% at 20 and 40 mg/mL, respectively. Gas chromatography-mass spectrometry (GC-MS) subjected this fraction to an acetylation reaction to determine the content of the secondary metabolites. The GC-MS analysis showed that the F5 fraction was composed of trehalose CAS: 6138-23-4, polyols (L-iditol CAS: 488-45-9, galactitol CAS: 608-66-2, D-mannitol CAS: 69-65-8, D-glucitol CAS: 50-70-4, and myoinositol CAS: 87-89-8), adipic acid CAS: 124-04-9, stearic acid CAS: 57-11-4, squalene CAS: 111-02-4, and ß-sitosterol CAS: 83-46-5.