Anaerobic digestion of spent mushroom substrate under thermophilic conditions: performance and microbial community analysis

Applied Microbiology and Biotechnology - Spent mushroom substrate (SMS) is the residue of edible mushroom production occurring in huge amounts. The SMS residue can be digested for biogas production...     

Broiler litter supplementation improves storage and feed-nutritional value of sawdust-based spent mushroom substrate

A study was conducted to determine the effect of broiler poultry litter (BL) supplementation to spent mushroom substrate (SMS) on its storage and feed-nutritional value improvement. In Exp. 1, the sawdust-based SMS from a king oyster mushroom (Pleurotus eryngii) farm was mixed with BL at 0%, 25%, 50%, 75% and 100% levels on a wet basis and deepstack stored for short-term (1-wk) and long-term (3-wk). At 1-wk of short-term deepstack storage, SMS with more than 50% BL levels showed favorable conservation. At 3-wk of long-term storage, all treatments except for BL 100% had a serious fungal problem. Based on chemical parameters, BL-blending to SMS practically improved the feed-nutritional value of the mixtures. Since the deepstacking method was not effective for long term storage, in Exp. 2 SMS ensiled with or without BL was attempted to improve long-term (3-wk) storage. All the ensiled treatments (SMS 100%, SMS 75%+BL 25% and SMS 50%+BL 50%) had desirable fermentation. As in deepstacking, BL-blending to SMS improved the nutritive value of the ensiled mixtures. The populations of total bacteria, lactic acid bacteria and yeast were highest (P<0.05) when 75% SMS was blended with 25% BL. In conclusion, blending 50% or more BL with SMS was effective for the short-term (1-wk) deepstack storage. For long-term (3-wk) storage of SMS, an ensiling method was effective with or without the addition of BL.     

Extraction, purification and antibacterial activities of a polysaccharide from spent mushroom substrate

To contribute towards effective exploitation and utilization of spent mushroom substrate (SMS), a water-soluble polysaccharide named PL was isolated and purified from SMS. The total sugar content and monosaccharide composition were analyzed by phenol-sulfuric acid method and capillary electrophoresis, and infrared spectroscopy was also performed for structure characterization. The results showed that the total sugar content of crude polysaccharide from SMS was about 25.8%, the polysaccharide contained two fractions (PL1 and PL2), which was mainly composed of glucose, rhamnose and mannose with a molar ratio of 1:3.13:1.16. The attributions of the main absorptions of both PL1 and PL2 were characteristic of glycosidic structures, and the FT-IR spectra of PL2 and lentinan were very similar. Escherichia coli, Staphylococcus aureus and Sarcina lutea were used to study the antibacterial activity and minimal inhibitory concentrations (MICs) of the polysaccharide. The antibacterial activity of polysaccharide from SMS against E. coli was the strongest, while the weakest against S. lutea, and the MICs of PL2 were 12.5, 25 and 100 μg/mL, respectively.     

The influence of bacterial inoculants on the microbial ecology of aerobic spoilage of barley silage.

The aerobic decomposition of barley silage treated with two inoculants (LacA and LacB) containing mixtures of Lactobacillus plantarum and Enterococcus faecium was investigated over a 28-day period. Initially, yeast and bacterial populations were larger in silage inoculated with LacA than in silage treated with LacB or water alone (control). Differences in the succession of yeasts in silage treated with LacA were observed relative to the other two treatments. From silage treatment with LacA, Issatchenkia orientalis was the most prevalent yeast taxon over all of the sample times, and the filamentous fungus Microascus brevicaulis was also frequently isolated at later sample dates (> or = 14 days). In contrast, Saccharomyces exiguus was the most prominent yeast recovered from silage treated with LacB and water alone on days 2 and 4, although it was supplanted by I. orientalis at later sample times. Successional trends of bacteria were similar for all three treatments. Lactobacillus spp. were initially the most prevalent bacteria isolated, followed by Bacillus spp. (primarily Bacillus pumilus). However, the onset of Bacillus spp. prominence was faster in LacA silage, and Klebsiella planticola was frequently recovered at later sample times (> or = 14 days). More filamentous fungi were recovered from LacA silage on media containing carboxylmethylcellulose, pectin, or xylan. The most commonly isolated taxa were Absidia sp., Aspergillus flavus, Aspergillus fumigatus, Byssochlamys nivea, Monascus ruber, Penicillium brevicompactum, Pseudoallescheria boydii, and M. brevicaulis. The results of this study indicated that the two bacterial inoculants incorporated into barley at the time of ensilage affected the microbial ecology of silage decomposition following exposure to air. However, neither of the microbial inoculants effectively delayed aerobic spoilage of barley silage, and the rate of decomposition of silage treated with one of the inoculants (LacA) was actually enhanced.     

Recycling of spent shiitake substrate for production of the oyster mushroom,Pleurotus sajor-caju

Pleurotus sajor-caju was produced on a basal medium containing spent shiitake substrate plus 10% wheat bran and 10% millet. An analysis of the fibrous composition of the spent shiitake substrate revealed that 85% of the original hemicellulose, 44% of the original cellulose and 77% of the original lignin was not consumed during production of a full crop (78% biological efficiency) of shiitake (63-day harvest period). To produce P. sajor-caju, the spent shiitake substrate was ground, air dried, supplemented, pasteurized with live steam and spawned. Highest yields (79% biological efficiency) of P. sajor-caju were obtained by supplementing the spent shiitake basal medium with 12% soybean and 1% CaCO₃. Increases in percentage biological efficiency and mushroom size were positively correlated with increasing levels of CaCO₃ added to the basal medium.     

Antibacterial activity of lactic acid bacteria included in inoculants for silage and in silages treated with these inoculants

AIMS: To determine antibacterial activity in lactic acid bacteria (LAB) silage inoculants and in wheat and corn silages which were treated with these inoculants. METHODS AND RESULTS: Wheat and two corn silages were prepared in 0.25 l sealed glass jars. Inoculant treatments were prepared for each type of silage with each of 10 LAB silage inoculants at inoculation rate of 10(6) CFU g(-1). Untreated silages served as controls. Antibacterial activity was determined in the inoculants and in their respective silages with Micrococcus luteus and Pseudomonas aeruginosa. Antibacterial activity was detected in nine of the 10 inoculants whereas such activity in the silages varied. Control silages did not have antibacterial activity. CONCLUSIONS: Many LAB silage inoculants have antibacterial activity and in some cases this activity is imparted on inoculated silages. SIGNIFICANCE AND IMPACT OF THE STUDY: This study was conducted as part of a broader research objective, which is to find out how LAB silage inoculants enhance ruminant performance. The results of this study indicate that LAB silage inoculants produce antibacterial activity, and therefore, have a potential to inhibit detrimental micro-organisms in the silage or in the rumen.     

Concepts and applications of foliar spray for microbial inoculants

Applied Microbiology and Biotechnology - Damages of the (agro)ecosystem by extensive use of chemical fertilizers and pesticides, the global dying of bee populations possibly linked to pesticide...     

Comparative analysis of amylolytic lactobacilli and Lactobacillus plantarum as potential silage inoculants

Abstract Two starch-degrading Lactobacillus strains, Lactobacillus amylovorus NRRLB4540 and Lactobacillus amylophilus NCIB11546, were assessed for their potential as silage inoculants. Lactobacillus amylovorus was considered suitable as a silage inoculant for crops which are low in water-soluble carbohydrate, but contain starch, which is unavailable to most conventional silage inoculants. This was on the basis of it exhibiting similar growth characteristics to an inoculant strain of Lactobacillus plantarum and secreting an amylase which was optimally active in the silage pH range. Lactobacillus amylophilus was found to be intolerant of low acid conditions but displayed considerable potential for the industrial production of lactic acid from inexpensive starch-based substrates as it fermented sugars to the desirable L(+) isomer of lactic acid only.     

New trends and opportunities in the development and use of inoculants for silage

Abstract: Inoculants are used as silage additives to improve preservation efficiency and to enhance animal performance. In most commercially available inoculants, homofermentative lactic acid bacteria (LAB) have been used because they are fast and efficient producers of lactic acid, improving natural silage fermentation. Specific LAB inuculants may also have beneficial effects on animal performance even if there is no effect on fermentation. However, these types of inoculants are not always advantageous. They do not necessarily prevent sermentation by clostridia in moist silages, and they sometimes impair the aerobic stability of grass and small grain silages. Therefore, new criteria for silage inoculants should be established which consider the specific needs of the crop being ensiled. New approaches which are being taken to develop improved inoculants for silage include the following: (1) using LAB isolates which are more specific to the target crops; (2) inclusion of heterofermentative LAB to produce volatile fatty acids to inhibit yeasts and moulds upon aerobic exposure; (3) inclusion of organisms other than LAB in inoculants to inhibit detrimental microorganisms; (4) selection or engineering of LAB strains to inhibit specific microorganisms; and (5) cloning and expression of genes which would enable selected LAB strains to utilize polysaccharides in crops which are low in soluble carbohydrates. Many of these new strategies for formulating inoculants are being tested, but further research is needed to determine the most successful approaches.     

Production of spent mushroom substrate hydrolysates useful for cultivation of Lactococcus lactis by dilute sulfuric acid, cellulase and xylanase treatment

Spent mushroom substrate (SMS) was treated with dilute sulfuric acid followed by cellulase and xylanase treatment to produce hydrolysates that could be used as the basis for media for the production of value added products. A L9 (3⁴) orthogonal experiment was performed to optimize the acid treatment process. Pretreatment with 6% (w/w) dilute sulfuric acid at 120 °C for 120 min provided the highest reducing sugar yield of 267.57 g/kg SMS. No furfural was detected in the hydrolysates. Exposure to 20 PFU of cellulase and 200 XU of xylanase per gram of pretreated SMS at 40 °C resulted in the release of 79.85 g/kg or reducing sugars per kg acid pretreated SMS. The dilute sulfuric acid could be recycled to process fresh SMS four times. SMS hydrolysates neutralized with ammonium hydroxide, sodium hydroxide, or calcium hydroxide could be used as the carbon source for cultivation of Lactococcus lactis subsp. lactis W28 and a cell density of 2.9 × 10¹¹ CFU/mL could be obtained. The results provide a foundation for the development of value-added products based on SMS.     

Influence of moisture content on microbial activity and silage quality during ensilage of food processing residues

Seasonally produced biomass such as sugar beet pulp (SBP) and tomato pomace (TP) needs to be stored properly to meet the demand of sustainable biofuel production industries. Ensilage was used to preserve the feedstock. The effect of moisture content (MC) on the performance of ensilage and the relationship between microorganism activities and MC were investigated. For SBP, MC levels investigated were 80, 55, 30, and 10% on a wet basis. For TP, MC levels investigated were 60, 45, 30, and 10%. Organic acids, ethanol, ammonia, pH and water soluble carbohydrates (WSC) were measured to evaluate the silage quality. Ensilage improved as the MC decreased from 80 to 55% for SBP and from 60 to 45% for TP. When the MC decreased to 30%, a little microbial activity was detected for both feedstocks. Storage at 10% MC prevented all the microbial activity. The naturally occurring microorganisms in TP were found to preserve TP during silage and were isolated and determined by polymerase chain reaction (PCR). The results suggest that partial drying followed by ensilage may be a good approach for stabilization of food processing residues for biofuels production.     

Yield and size of oyster mushroom grown on rice/wheat straw basal substrate supplemented with cotton seed hull

Oyster mushroom (Pleurotus ostreatus) was cultivated on rice straw basal substrate, wheat straw basal substrate, cotton seed hull basal substrate, and wheat straw or rice straw supplemented with different proportions (15%, 30%, and 45% in rice straw substrate, 20%, 30%, and 40% in wheat straw substrate) of cotton seed hull to find a cost effective substrate. The effect of autoclaved sterilized and non-sterilized substrate on growth and yield of oyster mushroom was also examined. Results indicated that for both sterilized substrate and non-sterilized substrate, oyster mushroom on rice straw and wheat basal substrate have faster mycelial growth rate, comparatively poor surface mycelial density, shorter total colonization period and days from bag opening to primordia formation, lower yield and biological efficiency, lower mushroom weight, longer stipe length and smaller cap diameter than that on cotton seed hull basal substrate. The addition of cotton seed hull to rice straw and wheat straw substrate slowed spawn running, primordial development and fruit body formation. However, increasing the amount of cotton seed hull can increase the uniformity and white of mycelium, yield and biological efficiency, and increase mushroom weight, enlarge cap diameter and shorten stipe length. Compared to the sterilized substrate, the non-sterilized substrate had comparatively higher mycelial growth rate, shorter total colonization period and days from bag opening to primordia formation. However, the non-sterilized substrate did not gave significantly higher mushroom yield and biological efficiency than the sterilized substrate, but some undesirable characteristics, i.e. smaller mushroom cap diameter and relatively long stipe length.     

Influence of formaldehyde-treated soybean and commercial nutrient supplementation on mushroom (Pleurotus sajor-caju) yield and in-vitro dry matter digestibility of spent substrate

Summary Pleurotus sajor-caju 537 was grown on chopped, pasteurized wheat straw non-supplemented and supplemented with formaldehyde-treated soybean, commercial delayed-release nutrient (SpawnMate II SE) or vegetable oil. Yield was 2.1-fold higher for substrate supplemented (12% dry wt) with low-volume formaldehyde-treated soybean as compared to non-supplemented substrate. Mushroom yield from substrate supplemented with commercial nutrient was 1.7-fold higher than yield from non-supplemented substrate. As the supplement level increased, the mushroom yield response increased. The yield ranged from 3.56 kg/m² for non-supplemented substrate to 7.36 kg/m² for substrate supplemented (12% dry wt) with formaldehyde-treated soybean. The type of supplement affected in vitro dry matter digestibility (IVDMD) of spent substrate; commercial supplement resulted in higher IVDMD compared to formaldehyde-treated substrate. An opportunity exists for commercial development of a nutrient(s) specifically designed for Pleurotus cultivation.     

促分解菌剂对还田玉米秸秆的分解效果及土壤微生物的影响

为了探明促分解菌剂的应用对还田玉米秸秆的促分解效果及对土壤微生物群落结构的影响,选用3组促分解菌剂,编号依次为ND、NK和NS,于2009年10月至2010年4月期间,在河北省农林科学院辛集实验站冬小麦-玉米轮作田对玉米秸秆还田地进行了接种试验。在接种后的15、25、145和160 d分别测定秸秆残重率和秸秆残渣中C/N,结果表明与未施菌剂对照(CK)相比,3组菌剂均在一定程度上加快了玉米秸秆的分解,其中以菌剂ND促分解效果最好,NK次之,NS较差,三者的最高促分解效果分别比CK提高了14.3%、7.7%和1.6%,主要促分解效果都出现在早期(前25 d),且菌剂促进秸秆残渣中C/N降低的效果也在早期明显。采用变性梯度凝胶电泳技术(DGGE)检测菌剂对玉米秸秆降解过程中土壤细菌和真菌群落结构的影响,结果表明,与不接种CK相比,接种菌剂主要在早期对土壤细菌和真菌的群落结构产生较大的影响,而后期对土壤微生物群的影响不明显。秸秆还田后接种促分解菌剂,能在接种早期有效加快秸秆分解,而随接种后时间的推进,其促进效果逐渐减弱,与之对应,土壤微生物群落结构早期差异明显,其后差异逐渐减小。     

Suppression of gray leaf spot (blast) of perennial ryegrass turf by Pseudomonas aeruginosa from spent mushroom substrate

Bacteria isolated from spent mushroom substrate (SMS) were evaluated for the suppression of Pyricularia grisea, the causal agent of gray leaf spot of perennial ryegrass (Lolium perenne) turf. Thirty-two of 849 bacterial isolates (3.8%) from SMS significantly inhibited the mycelial growth of P. grisea in vitro. Six bacterial isolates that afforded maximum inhibition of P. grisea were also refractory to Rhizoctonia solani, Rhizoctonia cerealis, Sclerotinia homoeocarpa, and Fusarium culmorum. Each of the six isolates was identified as Pseudomonas aeruginosa by fatty acid profile analysis. The biocontrol activity of the bacterial isolates was not compromised by a prolonged exposure to UV radiation in vitro. In controlled-environment chamber experiments, all 32 bacterial isolates were tested for suppression of gray leaf spot on Pennfine perennial ryegrass when applied as seed treatment or foliar sprays. Foliar applications of the bacteria (10⁸ cfu/ml 0.1% carboxymethylcellulose), but not the seed treatment, significantly reduced disease severity and incidence. The three most efficient isolates from foliar application treatments, which were among the six bacterial isolates identified as P. aeruginosa, were further evaluated for suppression of gray leaf spot as a function of timing of application. The three isolates of P. aeruginosa suppressed gray leaf spot in perennial ryegrass in Cone-tainers when applied at 1, 3, and 7 days prior to inoculation with P. grisea both in controlled-environment chamber experiments, and in potted ryegrass plants maintained in the field. All application intervals, regardless of the bacterial isolate, provided significant reduction of gray leaf spot severity. Suppression of gray leaf spot by isolates of P. aeruginosa under controlled-environment chamber conditions was not different from that observed in potted ryegrass plants maintained in the field. In field experiments, an isolate of P. aeruginosa provided significant suppression of gray leaf spot when applied at 1, 7, and 14 days prior to inoculation with P. grisea. The bacterium proved effective against gray leaf spot of perennial ryegrass maintained as fairway and rough heights. These results indicate that P. aeruginosa may be a potential biocontrol agent for gray leaf spot of perennial ryegrass turf.     

Effect of spent mushroom compost tea on mycelial growth and yield of button mushroom (Agaricus bisporus)

Preliminary studies suggested that the use of compost tea made from spent mushroom substrate (SMS) may be regarded as a potential method for biologically controlling dry bubble disease in button mushroom. The aim of this study was to assess the effect of SMS compost tea on the host, the button mushroom, to ascertain whether the addition of these water extracts has a toxic effect on Agaricus bisporus mycelium growth and on mushroom yield. In vitro experiments showed that the addition of SMS compost tea to the culture medium inoculated with a mushroom spawn grain did not have an inhibitory effect on A. bisporus mycelial growth. The effect of compost teas on the quantitative production parameters of A. bisporus (yield, unitary weight, biological efficiency and earliness) was tested in a cropping trial, applying the compost teas to the casing in three different drench applications. Quantitative production parameters were not significantly affected by the compost tea treatments although there was a slight delay of 0.8-1.4 days in the harvest time of the first flush. These results suggest that compost teas have no fungitoxic effect on A. bisporus so that they can be considered a suitable biocontrol substance for the control of dry bubble disease.     

Yield,size, and mushroom solids content of <Emphasis Type="Italic">Agaricus bisporus</Emphasis> produced on non-composted substrate and spent mushroom compost

Three crops of Agaricus bisporus were grown on non-composted substrate (NCS), spent mushroom compost (SMC), a 50/50 mixture of NSC/SMC, or pasteurized Phase II compost. NCS consisted of oak sawdust (28% oven dry wt), millet (29%), rye (8%), peat (8%), ground alfalfa (4%), ground soybean (4%), wheat bran (9%) and CaCO₃ (10%). Substrates were non-supplemented or supplemented with Target^® (a commercial delayed release nutrient for mushroom culture) or soybean meal at spawning or casing, or with Micromax^® (a mixture of nine micronutrients) at spawning. Mushroom yield (27.2 kg/m²) was greatest on a 50/50 mixture of NCS/SMC supplemented with 10% (dry wt) Target^® at casing. The same substrate supplemented with Target^® at spawning yielded 20.1 kg/m². By comparison, mushroom yield on Phase II compost supplemented at casing or at spawning with Target^® was 21.6 kg/m² and 20.6 kg/m², respectively. On NCS amended with 0.74% or 0.9% Micromax^® at spawning, yields increased by 51.8% (12.9 kg/m²) and 71.8% (14.6 kg/m²), respectively, over non-amended NCS (8.5 kg/m²). Conversely, mushroom yields were not affected when Micromax^® was added to a 50/50 mixture of NCS/SMC. Mushroom solids content was higher in mushrooms harvested from NCS amended with 0.74% Micromax^® (9.6%) compared to non-amended NCS (8.3%).