Bio-directed Chemical Study of Pleurotus ostreatus Spent Substrate and Its Nematicidal Activity.

PURPOSE: In the present study, the nematicidal activity of the chemical fractionation of the spent substrate of the edible mushroom Pleurotus ostreatus against eggs and L3 larvae of Haemonchus contortus was evaluated. METHODS: The hydroalcoholic extract of the spent substrate was subjected to a bipartition with ethyl acetate giving two fractions: one aqueous (F. Ac) and one organic (F. AcOET). Both fractions were evaluated against eggs and L3 larvae at different concentrations (5, 2.5, 1.25, 0.625 and 0.3125 mg/mL) and 2% methanol, PBS and thiabendazole (5 mg/mL) as controls. Chemical fractionation of F. AcOET was performed in open column chromatography where 76 fractions were obtained and when analyzed by thin layer chromatography (TLC) were grouped into 11 mixtures (R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11). These mixtures were evaluated at 10 mg/mL against eggs and L3 larvae. The data obtained were analyzed by ANOVA and a Tukey test in the SAS V9 program. RESULTS: The results showed that, in the evaluation of F. AcOET and F. Ac, for the inhibition of egg hatching, the highest percentages were 78.80 and 76.89% at 5 mg/mL, respectively. As for the percentage of larval mortality, F. AcOET obtained 60.91 and F. Ac 29.77% at 5 mg/mL. The results of the evaluations of the mixtures showed that mixtures R4, R5 obtained 100 and 95.41% larval mortality and mixtures R6 and R7 presented 100% inhibitory activity of egg hatching, so these were analyzed by gas chromatography mass spectrometry finding compounds such as vanillin, ß-sitosterol, ρ-methyl ρ-hydroxycinnamate and ρ-hydroxybenzaldehyde. CONCLUSION: The results of the present study demonstrate that the spent substrate of P. ostreatus has potential anthelmintic activity against H. contortus. Moreover, by reusing and taking advantage of this substrate, its environmental pollution effects can be reduced.

The Edible Mushroom Pleurotus djamor Produces Metabolites with Lethal Activity Against the Parasitic Nematode Haemonchus contortus.

Pleurotus djamor is an edible mushroom that has medicinal properties. This study aimed to assess the in vitro activity of P. djamor extracts and fractions against Haemonchus contortus eggs and exsheathed infective larvae (L3). Crude hydroalcoholic extracts were obtained by maceration and fractions were obtained through chromatography. Metabolite identity was determined using gas chromatography/mass spectrometry (GC-MS) analysis. The results showed that P. djamor extracts had no significant activity against eggs at the concentrations used. However, the extract showed 98.7% and 77% larval mortality 72 h postconfrontation at 320 and at 160 mg/mL, respectively. The chromatography analysis resulted in 23 fractions that were eventually grouped into three fractions (E1, E2, and E3). These fractions showed the following egg hatching inhibition percentages: E1 = 100, E2 = 38.7, and E3 = 5.5 at 10 mg/mL concentration 72 h postexposure. Likewise, larval mortality percentages after this period were 90.6, 100, and 0.44 at 40 mg/mL (P < .05), respectively. The GC-MS showed five major compounds in E1 fraction, including four fatty acids: (i) pentadecanoic, (ii) hexadecanoic, (iii) octadecadienoic, (iv) octadecanoic acid, and one terpene identified as ß-sitosterol. We concluded that the edible mushroom P. djamor possesses nematicidal metabolites, which could be used as an alternative anthelmintic treatment.