Dietary Agaricus blazei Spent Substrate Improves Disease Resistance of Nile Tilapia (Oreochromis niloticus) against Streptococcus agalactiae In Vivo

This study evaluated the effects of the feeding of spent mushroom substrate from Agaricus blazei on Nile tilapia (Oreochromis niloticus). The safety of 0–1000 μg/mL A. blazei spent substrate water extract (ABSSE) was demonstrated in the primary hepatic and splenic macrophages and the THK cell line (a cell line with characteristics of melanomacrophages) using a cytotoxicity assay. Here, 10 μg/mL of crude ABSSE promoted the phagocytic activity of macrophages and THK cells. Stimulating ABSSE-primed THK cells with lipopolysaccharides or peptidoglycan resulted in higher expression levels of four cytokine genes (e.g., interleukinz (IL)-1β, IL-12b, IL-8 and tumor necrosis factor α (TNFα)) and one cytokine gene (TNFα), respectively. An in vitro bacterial growth inhibition assay demonstrated that ABSSE could inhibit the growth of Streptococcus agalactiae. In the first feeding trial, Nile tilapia were fed with experimental feed containing 0, 1, or 5% of A. blazei spent substrate (ABSS) for seven and fourteen days followed by bacterial challenge assay. The best result was obtained when Nile tilapia were continuously fed for seven days on a diet containing 1% ABSS, with the survival rate being higher than in groups with 0% and 5% ABSS after challenge with S. agalactiae. In the second trial, fish were fed diets supplemented with 0% or 1% ABSS for seven days, and then all the groups were given the control feed for several days prior to bacterial challenge in order to investigate the duration of the protective effect provided by ABSS. The results showed that the protective effects were sustained at day 7 after the feed was switched. Overall, spent mushroom substrate from A. blazei is a cost-effective feed additive for Nile tilapia that protects fish from S. agalactiae infection.

Ascomycetes versus Spent Mushroom Substrate in Mycoremediation of Dredged Sediments Contaminated by Total Petroleum Hydrocarbons: The Involvement of the Bacterial Metabolism

Two mycoremediation approaches for the depletion of the total petroleum hydrocarbons in dredged sediments were compared: co-composting with spent mushroom substrate (SMS) from Pleurotus ostreatus and bioaugmentation with Lambertella sp. MUT 5852, an ascomycetes autochthonous to the sediment, capable of utilizing diesel oil its sole carbon source. After 28 days of incubation, 99% depletion was observed in presence of Lambertella sp. MUT 5852. No total petroleum hydrocarbon depletion was observed in sediment co-composting with the SMS after 60 days of incubation. 16S rDNA metabarcoding of the bacterial community was performed to evaluate the potential synergism between fungi and bacteria in the bioremediation process. The functional metagenomic prediction approach indicated that the biodiversity of the bacterial genera potentially involved in the degradation of TPH was higher in sediment bioaugmented with Lambertella sp. MUT 5852, which resulted in being mandatory for TPH depletion. Mechanisms of co-substrate inhibition of the hydrocarburoclastic bacterial species, due to the bioavailable organic matter of the SMS, are suggested to be involved in the observed kinetics of TPH depletion, failing in the case of SMS and successful in the case of Lambertella sp. MUT 5852.

Recovery of Lignocellulolytic Enzymes and Valorization of Spent Mushroom Substrate

Spent Mushroom Substrate (SMS) comprises sugarcane bagasse, coconut coir, chicken manure, and paddy straw; inoculated with and farmed for Agaricus bisporus. At present, the waste generation at a mushroom cultivation plant in Goa is 40 tons/day (15,000 tons annually). Valorization of this waste has been explored in terms of extracting lignocellulolytic enzymes and briquette production. SMS was screened for the presence of lignocellulolytic enzymes and then was used to make briquettes. The enzymes found in SMS were cellulase and laccase, which were further concentrated via tangential flow filtration (TFF). Enzyme activity for Cellulase increased by four-fold (from 255.34±1.30 U/mL increased to 1022.21±4.84 U/mL) and Laccase increased by three-fold (from 4.83±0.02 U/mL to 13.21±0.05 U/mL). The concentrated enzyme cocktail was used to decolorize congo red dye. After only eight hours of enzymatic treatment at pH 4.8 on congo red, approx. 40-49% decolorization was accomplished. The color removal was due to the presence of the laccase enzyme. After enzyme extraction, all the residual SMS was utilized to generate briquettes with an initial reduction in its moisture content from 50% to 10%. The resulting briquette gave a Gross Calorific Value of 4,143 Kcal/kg with 12.60% ash content. Thus, SMS proves to be a valuable source for recovering enzymes and a cost-effective material for briquette production rather than going into landfills.

Planifilum fulgidum Is the Dominant Functional Microorganism in Compost Containing Spent Mushroom Substrate

The extensive accumulation of spent mushroom substrate (SMS) owing to the large-scale production of edible fungi is causing environmental problems that cannot be ignored. Co-composting is a promising method for agricultural and animal husbandry waste disposal. In this study, the composition and function of microbial communities in the process of cattle manure–maize straw composting with SMS addition were compared through an integrated meta-omics approach. The results showed that irrespective of SMS addition, the predominant fungi were Ascomycota, while the dominant bacteria were Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes. High temperature promoted the evolution from Gram-negative bacteria (Bacteroides, Proteobacteria) to Gram-positive bacteria (Firmicutes, Actinomycetes). The composting process was accelerated by SMS addition, and the substrate was effectively degraded in 14 days. Metaproteomics results showed that the dominant microorganism, Planifilum fulgidum, secreted large amounts of S8, M17, and M32 proteases that could degrade macromolecular protein substrates in the presence of SMS. Planifilum fulgidum, along with Thermobifida fusca and Melanocarpus albomyces, synergistically degraded hemicellulose, cellulose, and protein. In addition, the dominant microorganisms related to the initial raw materials such as Pichia, Lactobacillus in the microbial agent and Hypsizygus in SMS could not adapt to the high-temperature environment (>60 °C) and were replaced by thermophilic bacteria after 5 days of composting.

SPENT MUSHROOM SUBSTRATE of Pleurotus ostreatus KUMMER INCREASES BASIL BIOMASS AND ESSENTIAL OIL YIELD

ABSTRACT Lignocellulosic residues are used to produce mushrooms, but they generate large amounts of spent mushroom substrate (SMS). The objective of this study was to evaluate they effect of SMS of Pleurotus ostreatus, combined with organic fertilization, on biomass production and essential oil yield of basil (Ocimum basilicum L.) plants. The fertilizer was formulated using combinations of organic compounds: SMS, organic compost (OC), and earthworm humus (EH). The treatments were applied using 35 g of the formulations, corresponding to an application of 20 Mg ha-1. The treatments used were: 100% OC; 100% EH; 100% SMS; 75% SMS + 25% OC (C1); 75% SMS + 25% EH (C2); 50% SMS + 50% OC (C3); 50% SMS + 50% EH (C4); 50% SMS + 25% OC + 25% EH (C5), and a control with no fertilizer application (CT). Plants grown with the soil fertilizers containing SMS, single or combined with OC and EH, presented higher average plant height (55.6 to 62.2 cm) and leaf area per plant (696.5 to 836.4 cm2). The treatment C3 resulted in plants with higher mean dry weight (10.9 g plant-1), and C2 resulted in the highest oil yield (5.0 kg ha-1), representing a gain of 324.8% in oil yield when compared to the control. Thus, SMS increases biomass production and essential oil yield of basil plants.