Use of spent mushroom substrates from Agaricus subrufescens (syn. A. blazei, A. brasiliensis) and Lentinula edodes productions in the enrichment of a soil-based potting media for lettuce (Lactuca sativa) cultivation: Growth promotion and soil bioremediation

This study aimed to assess physicochemical and microbiological properties of fresh spent mushroom substrates (SMSs) – without post-crop heat treatment – from Agaricus subrufescens and Lentinula edodes production to optimize the use of these residues in the soil enrichment for lettuce growth promotion and soil remediation. Organic matter and C content of both SMSs were high. Fresh A. subrufescens SMS was a good source of N, P and K. On the other hand, L. edodes SMS presented a lower concentration of these nutrients and a high level of immaturity. Both SMSs presented high electric conductivity values (2.5–3.4 mS/cm). Microbiological analysis, based upon enumeration of culturable bacteria (thermophilic and mesophilic) and fungi, and also evolution of CO₂, showed that SMSs played higher microbial diversity than soil control. Laccase activity from A. subrufescens SMS tended to remain constant during a 2-month period, while L. edodes SMS presented low laccase activity throughout the same period. Agaricus subrufescens and L. edodes were able to grow on a PDA (Potato Dextrose Agar) media supplemented with different concentrations of atrazine (1–50 μg/ml), degraded the herbicide, attaining rates of 35% and 26%, respectively. On experiments of lettuce growth promotion using a soil-based potting media with different SMS rates, 5% and 10% (dw) rates of A. subrufescens SMS resulted in higher lettuce aerial dry weights than the rates of 25% and 40%, the chemical fertilization (NPK) and the control (soil). At 10% supplementation, lettuce aerial dry weight increased 2.2 and 1.3 times compared to the control and the NPK treatment, respectively. Protein content increased along with SMS rates. Fresh A. subrufescens SMS was an excellent supplement for lettuce growth promotion and showed potential for remediation of biocides possibly due to improved microbial diversity and enzymatic activity. Fresh L. edodes SMS was not a good fertilizer, at least under the conditions tested. However, microbiological analysis showed that promising results may be achieved when using fresh L. edodes SMS for soil remediation.     

Comparative assessment of chelated spent mushroom substrates as casing material for the production of Agaricus bisporus

Spent mushroom substrate (SMS) leached with water or treated with chelating agents to remove metal cations, pasteurised to remove any harmful micro-organisms and mixed with peat has potential as a casing material for mushroom production. The microbial and chemical changes in SMS after treatment with citric acid, ethylene diaminetetraacetic acid (EDTA) and water were compared; treatment with the chelating agents resulted in lower ash content, conductivity and minerals, higher fibre fractions, carbon, hydrogen and nitrogen. The microbial and chemical changes in the materials after treatment with the two chelators and water were compared. Blending peat with the heat-treated materials at a ratio of 1:1 resulted in improved physical properties. The casings prepared from the test materials and the control, consisting of 100% peat, were compared after neutralising with lime for their productivity in a mushroom yield trial. As expected, the compost bags cased with the control were the most productive compared to the other casings. Of the three treated materials, casing prepared from SMS treated with EDTA blended with peat was the most productive. Dry matter of harvested mushrooms from chelated-SMS casings was significantly higher than the control casing. Comparison of the main components of peat and chelated SMS revealed that the major differences were in the proportions of ash, lipid, lignin and fibre fractions. The stability of some of these components, when complexed with metal cations present in lime may play an important role in determining the composition of the cell wall in fruiting bodies leading to high dry matter content. Received: 19 November 1998 / Received revision: 29 March 1999 / Accepted: 6 April 1999     

Improving biocontrol of black vine weevil (<Emphasis Type="Italic">Otiorhynchus sulcatus</Emphasis>) with entomopathogenic fungi in growing media by incorporating spent mushroom compost

Amending a peat-based growing medium with 10% v/v spent mushroom compost, a source of fungal chitin and other nutrients, prolonged the persistence of entomopathogenic fungi (Metarhizium brunneum Petsch and Beauveria bassiana (Balsamo) Vuillemin; Hypocreales: Clavicipitaceae). This resulted in improved efficacy of M. brunneum against black vine weevil, Otiorhynchus sulcatus F. (Coleoptera: Curculionidae) larvae compared with using inoculum without spent mushroom compost. B. bassiana only controlled larvae when used in combination with spent mushroom compost (75?±?7% reduction in live larvae). Mixing entomopathogenic fungal inoculum with spent mushroom compost and growing medium was as effective in controlling black vine weevil larvae as using spent mushroom compost colonised with M. brunneum or B. bassiana in the growing medium (80?±?12% reduction in live larvae). The former method is preferable since it does not require production and storage of colonised spent mushroom compost, or registration of new substrate formulations of M. brunneum or B. bassiana.     

Germination and biometric parameters of seedlings grown on solid pulp and paper waste medium

Germination and biometric parameters (number and length of needles, root length, trunk height, root neck diameter, and fractional seedling mass) of pine (Pinus sylvestris L.) and spruce (Picea abies L. Karst) seedlings with a closed root system are evaluated. The seedlings are grown on a mix of peat with growing media prepared of pulp and paper mill waste (PPW) conversed by worms (Eisenia foctida) using chlorine-free pulp bleaching technology. The growing media includes active sludge; mixes of soil, bark, and woodchip product slivers ground together in a crusher; and coniferous sawdust. The share of (PPW) media is 20, 50 and 100 vol %. The experimental results have shown that in most cases the parameters of pine and spruce seedlings grown on these media in small volumes (20%) reach the parameters of the control samples represented by the seedlings grown on fertilized peat. When a growing medium is used in small amounts, the aboveground/subterraneous ratio of the seedlings is also similar to the control values. The more the growing medium is applied, the lower the parameters of the seedlings are in comparison with the control figures. This is probably caused by the offset of the nitrogen status of the growing media to nitrate nitrogen, which is not typical of natural forest soils.     

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.     

Determination of absolute coccolith abundances in deep-sea sediments by spiking with microbeads and spraying (SMS-method)

A quick new method is described for the quantification of absolute nannofossil proportions in deep-sea sediments. This method (SMS) is the combination of Spiking a sample with Microbeads and Spraying it on a cover slide. It is suitable for scanning electron microscope (SEM) analyses and for light microscope (LM) analyses. Repeated preparation and counting of the same sample (30 times) revealed a standard deviation of ±10.5%. The application of tracer microbeads with different diameters and densities revealed no statistically significant differences between counts. The SMS-method yielded coccolith numbers that are statistically not significantly different from values obtained from the filtration-method. However, coccolith counts obtained by the random settling method are three times higher than the values obtained by the SMS- and the filtration-method.     

Mutants of the pentose-fermenting yeast Pachysolen tannophilus tolerant to hardwood spent sulfite liquor and acetic acid

A strain development program was initiated to improve the tolerance of the pentose-fermenting yeast Pachysolen tannophilus to inhibitors in lignocellulosic hydrolysates. Several rounds of UV mutagenesis followed by screening were used to select for mutants of P. tannophilus NRRL Y2460 with improved tolerance to hardwood spent sulfite liquor (HW SSL) and acetic acid in separate selection lines. The wild type (WT) strain grew in 50 % (v/v) HW SSL while third round HW SSL mutants (designated UHW301, UHW302 and UHW303) grew in 60 % (v/v) HW SSL, with two of these isolates (UHW302 and UHW303) being viable and growing, respectively, in 70 % (v/v) HW SSL. In defined liquid media containing acetic acid, the WT strain grew in 0.70 % (w/v) acetic acid, while third round acetic acid mutants (designated UAA301, UAA302 and UAA303) grew in 0.80 % (w/v) acetic acid, with one isolate (UAA302) growing in 0.90 % (w/v) acetic acid. Cross-tolerance of HW SSL-tolerant mutants to acetic acid and vice versa was observed with UHW303 able to grow in 0.90 % (w/v) acetic acid and UAA302 growing in 60 % (v/v) HW SSL. The UV-induced mutants retained the ability to ferment glucose and xylose to ethanol in defined media. These mutants of P. tannophilus are of considerable interest for bioconversion of the sugars in lignocellulosic hydrolysates to ethanol.     

Composted Green Waste as a Substitute for Peat in Growth Media: Effects on Growth and Nutrition of Calathea insignis

Peat mined from endangered wetland ecosystems is generally used as a component in soilless potting media in horticulture but is a costly and non-renewable natural resource. The objective of this work was to study the feasibility of replacing peat with different percentages (0, 10, 30, 50, 70, 90, and 100%) of composted green waste (CGW) as growth media for the production of the ornamental plant Calathea insignis. Compared with 100% peat media, media containing CGW had improved physical and chemical characteristics to achieve the acceptable ranges. Moreover, CGW addition had increased the stability (i.e., reduced the decomposition rates) of growth media mixtures, as indicated by comparison of particle-size distribution at the start and end of a 7-month greenhouse experiment. Addition of CGW also supported increased plant growth (biomass production, root morphology, nutrient contents, and photosynthetic pigment contents). The physical and chemical characteristics of growth media and plant growth were best with a medium containing 70% CGW and were better in a medium with 100% CGW than in one with 100% peat media. These results indicate that CGW is a viable alternative to peat for the cultivation of Calathea insignis.     

A laboratory evaluation of the pathogenicity of Bacillus popilliae var. rhopaea, the agent of milky disease in Rhopaea verreauxi (Coleoptera: Scarabaeidae)

Two methods of infection, i.e., feeding known numbers of spores and rearing larvae in contaminated peat, were used to bioassay the susceptibility of Rhopaea verreauxi to Bacillus popilliae var. rhopaea at 23°C. The susceptibility of the three larval instars was similar as measured by the ID₅₀ and IC₅₀ values. However, within an instar, newly molted larvae were less susceptible than mature larvae when infected by the contaminated peat method. It is suggested that this was due to reduced food intake. The range of ID₅₀ values for all bioassays with R. verreauxi larvae were 1.1 × 10⁷ to 4.0 × 10⁷ spores per larva, and IC₅₀ values were 3.4 × 10⁶ to 5.0 × 10⁷ spores per g of contaminated peat. The slope of the probit line was always low (0.6 to 1.8) except for young first-instar larvae infected by contaminated peat when the slope was 4.0. Disease per se did not affect food intake, though intake was reduced at high doses of contaminated peat. Young larvae often died without developing symptoms but, with increasing age, infected larvae were more likely to develop symptoms. Bioassays with Othnonius batesi and Rhopaea morbillosa indicated a much lower susceptibility per os than for R. verreauxi. It is concluded that the potential for using B. popilliae var. rhopaea to control R. verreauxi is high, but the bacillus is unlikely to be of value in control of O. batesi or R. morbillosa.     

Use of composted sewage sludge in growth media for broccoli

In this study, the use of composted sewage sludge (CSS) as a binary component with peat (P) in growth media for a horticultural crop, broccoli (Brassica oleracea var. Botryti cv. Marathon), was evaluated. Four treatments were established, based on the addition of increasing quantities of composted sewage sludge to peat (0%, 15%, 30% and 50%, v/v). Physical, physico-chemical and chemical analyses of the different mixtures of CSS and P were made. Plant growth, biomass production and macronutrient (N, P, K, Ca, Mg), micronutrient (Fe, Cu, Mn, Zn) and heavy metal (Pb, Ni, Cd, Cr) contents of plants were determined. The addition of CSS to P increased plant nutrient and heavy metal contents of plants and electrical conductivity (EC) and bulk density values of the substrates. The use of CSS did not affect the germination rate, even at 50% compost. For broccoli growth, the highest yield was obtained with the medium prepared by mixing the peat with 30% of compost; however, the mixture with the most sewage sludge compost (50%) had the greatest contents of macro and micronutrients.     

The Cultivation of Arabidopsis for Experimental Research Using Commercially Available Peat-Based and Peat-Free Growing Media

Experimental research involving Arabidopsis thaliana often involves the quantification of phenotypic traits during cultivation on compost or other growing media. Many commercially-available growing media contain peat, but peat extraction is not sustainable due to its very slow rate of formation. Moreover, peat extraction reduces peatland biodiversity and releases stored carbon and methane into the atmosphere. Here, we compared the experimental performance of Arabidopsis on peat-based and several types of commercially-available peat-free growing media (variously formed from coir, composted bark, wood-fibre, and domestic compost), to provide guidance for reducing peat use in plant sciences research with Arabidopsis. Arabidopsis biomass accumulation and seed yield were reduced by cultivation on several types of peat-free growing media. Arabidopsis performed extremely poorly on coir alone, presumably because this medium was completely nitrate-free. Some peat-free growing media were more susceptible to fungal contamination. We found that autoclaving of control (peat-based) growing media had no effect upon any physiological parameters that we examined, compared with non-autoclaved control growing media, under our experimental conditions. Overall, we conclude that Arabidopsis performs best when cultivated on peat-based growing media because seed yield was almost always reduced when peat-free media were used. This may be because standard laboratory protocols and growth conditions for Arabidopsis are optimized for peat-based media. However, during the vegetative growth phase several phenotypic traits were comparable between plants cultivated on peat-based and some peat-free media, suggesting that under certain circumstances peat-free media can be suitable for phenotypic analysis of Arabidopsis.     

Physiologische und biochemische Beiträge zur Taxonomie der GattungChlorella

Salt tolerance was found to be a species-specific character in the genus Chlorella. The most resistant species, C. luteoviridis, is able to grow in nutrient media containing up to 5% NaCl. The limit for C. protothecoides and C. saccharophila is at 4% NaCl. C. vulgaris var. vulgaris, C. fusa var. vacuolata and C. fusca var. rubescens grow in the presence of 3%, and C. fusca var. fusca, C. kessleri and C. vulgaris f. tertia in the presence of 2% NaCl. C. zofingiensis and C. minutissima are only capable of growing in media containing up to 1% NaCl, and C. homosphaera does not even tolerate 1% NaCl. Those strains, which are able to synthesize secondary carotenoids, turn orange in salt concentrations close to the limit of their tolerance.     

Somatostatin does not attenuate intestinal injury in dextran sodium sulphate-induced subacute colitis.

FRom several in vitro and in vivo studies involvement of somatostatin (SMS) in intestinal inflammation emerge. Acute colitis induced in rats is attenuated by the long-acting SMS analogue octreotide. We studied the potential beneficial effect of SMS on non-acute experimental colitis. BALB/c mice received either saline, SMS-14 (36 or 120 microg daily) or octreotide (3 microg daily) subcutaneously delivered by implant osmotic pumps. A non-acute colitis was induced by administration of dextran sodium sulphate (DSS) 10% in drinking water during 7 days. DSS evoked a mild, superficial pancolitis, most characterized by mucosal ulceration and submucosal influx of neutrophils. Neither SMS-14 nor octreotide reduced mucosal inflammatory score or macroscopical disease activity, although reduction of intestinal levels of interleukin-1beta (IL-1beta), IL-6 and IL-10 during DSS was augmented both by SMS and octreotide. A slight increase of neutrophil influx was seen during SMS administration in animals not exposed to DSS. In conclusion, SMS or its long-acting analogue did not reduce intestinal inflammation in non-acute DSS-induced colitis. According to the cytokine profile observed, SMS-14 and octreotide further diminished the reduction of intestinal macrophage and Th2 lymphocyte activity.     

Influence of Ficus carica and Olea europaea leaves extracts on the mycelial growth of mushrooms in vitro

The use of 20% plant leaves extracts included fig (Ficus carica) and olive (Olea europaea) and their mixture 1:1 as an amendment in the solid agar medium (PDA) is beneficial to promote the growth of four mycelial mushrooms. These are Pleurotus ostreatus (Grey oyster mushroom), Pleurotus cornucopiae (Yellow oyster mushroom), Coriolus versicolor (Turkey Tail mushroom), and Ganoderma lucidum (Reishi mushroom). C. versicolor showed better growth reached 67?mm significantly (p?<?0.05) on OC medium after five days. While, P. cornucopiae recorded the lowest growth on FC medium reached 35.3?mm. Induction percentage of mycelial growth is changing according to the type of medium and species of fungus. In general, FOH medium exhibited the best percentage of induction was 14.89%, followed 12.48% and 9.43% by OH and OC media, while the lower percentages were 5.02% and 5.12% on FH and FC media, respectively. FC medium did not induce growth of P. cornucopiae and C. versicolor. The sterilization by Autoclave and Millipore filter showed different induction percentages. Finally, the extracts of fig and olive were useful to add in the culture media to improve the growth of mycelial mushroom in vitro.     

Pollinator visitation to mass-flowering courgette and co-flowering wild flowers: Implications for pollination and bee conservation on farms

Managing the complex relationship between pollinators and their habitat requirements is of particular concern to growers of pollinator-dependent crop species, such as courgette (Cucurbita pepo). Naturally occurring wild flowers (i.e. agricultural weeds) offer a free, sustainable, and often underappreciated resource for pollinators, however, they may compete with crop flowers for visits. To understand the extent to which floral resources mediate pollinator visitation to courgette flowers and courgette fields, plant community and pollinator visitation data were collected at two spatial scales: field scale (in margins, and in the cropped area) and farm scale (500 m and 2000 m radii) for nine courgette fields across the UK. Apis mellifera (honeybees) and Bombus spp. (bumblebees) were the only pollinators observed to visit courgette flowers. Bumblebees were significantly more abundant on courgette flowers in fields with a greater species richness of wild flowers in the crop, whilst honeybees were significantly more abundant on courgette flowers in areas with less semi-natural habitat. For both honeybees and bumblebees, their abundance in field margins did not significantly reduce their abundance on courgette flowers, suggesting that wild flowers were not competing with courgette flowers for pollinator visitation. Although solitary bees were not observed to visit courgette flowers, their abundance and species richness in courgette fields were significantly greater with more semi-natural habitat and a greater species richness of wild flowers. Therefore, allowing uncultivated areas around the crop to be colonised by species-rich wild flowers is an effective way of boosting the abundance of bumblebees, which are important visitors to courgette flowers, as well as the abundance and species richness of solitary bees, thereby benefitting pollinator conservation.     

Properties of peat-based casing soils and their influence on the water relations and growth of the mushroom (Agaricus bisporus)

Different combinations of peat and chalk or lime sources with differing moisture contents were used to determine how specific physical and chemical properties of the casing soil relate to the growth and water relations of the mushroom. The peat types varied in terms of decomposition and extraction method; the lime addition varied in terms of rate and type (chalk or sugar beet lime). During the colonisation of the casing soil before fruiting, the extension growth rate of mushroom mycelium was most closely correlated (negatively) with the volumetric moisture content of the casing soil. Scanning electron microscopy showed that mycelium growing at a lower casing soil matric potential (Ψ_m) had a much finer and branched structure than mycelium growing at a higher Ψ_m. Across all the peat and lime source treatments, a relationship was found between the mean Ψ_m of the casing soil and mushroom yield, with an optimum Ψ_m of -7.9 to -9.4 kPa. Mushrooms are produced in ‘flushes’ at about 8-day intervals and during the development of each flush of mushrooms, there was a significant decrease in casing soil Ψ_m . This decrease (to below -40 kPa) was greatest in the second flush, which was the highest yielding. There were no relationships between mushroom yield and casing soil osmotic potential Ψ_π within the range -93 to -154 kPa or any of the other chemical properties and water and air holding characteristics of the casing soils which were determined. Across different casing soil treatments, mushroom dry matter content was negatively correlated with mushroom yield and positively correlated with mushroom tissue osmotic potential. This revised version was published online in June 2006 with corrections to the Cover Date.     

Salt tolerance of Agaricus bisporus in relation to water stress and toxicity of sodium ions

Using NaCl or polyethylene glycol (PEG) solutions to progressively decrease the external osmotic potential of the peat casing of the growing medium used to culture the mushroom Agaricus bisporus resulted in proportionately decreased yields of sporophores. Over the range of -0.07 to -0.37 MPa, the extent of decrease in yield was similar with both types of osmoticum. However, with further decrease in external osmotic potential (from -0.37 to -0.62 MPa) there was a further proportional decrease in sporophore yield with PEG but a complete suppression of sporophore production with NaCl. Treatments with both NaCl and PEG decreased the concentrations of P, Mg, K, Fe and Mn, but not N and Cu, in sporophore dry matter. Treatment with NaCl solutions increased the concentrations of Na and CI ions in sporophore dry matter and decreased the concentration of Ca; PEG solutions had no effect. Ion toxicity associated with excessive accumulation of Na and C1 ions, or ionic imbalance associated with the concomittant decrease in Ca ions appear to be additional factors to osmotic stress in decreasing yield of sporophores when the growing medium becomes highly saline. The critical concentration of NaCl which caused 10% reduction in sporophore yield was 28 mM; A. bisporus is, therefore, moderately salt-sensitive.     

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.     

Growth of ornamental plants in two composts prepared from agroindustrial wastes

Two composts prepared from agroindustrial wastes were assayed as substrates: C1 from brewing waste (yeast and malt) plus lemon tree prunings; and C2 from the solid fraction of olive mill wastewater plus olive leaves. Sixteen substrates were prepared by combining each compost with Sphagnum peat or a commercial substrate (CS) in different proportions. The nutrients (N and K) provided by the composts, which acted as slow-release fertilisers, influenced especially the development of calendula, although the physical and physico-chemical properties such as total pore space and electrical conductivity (EC) were also relevant. On the other hand, in the salt-sensitive calceolaria hybrid, EC and chloride concentration were the main factors influencing growth. Adequate substrates for the development of calendula can be prepared by mixing C1 at up to 75% with peat or at up to 50% with CS, and C2 at up to 50% with peat or CS. For calceolaria, the substrate should have a lower proportion of compost, C1 at up to 50% and C2 at up to 25%, both mixed with peat or CS. Therefore, composts of agroindustrial origin such as these can be used as an alternative to peat and CSs for growing ornamental plants. provided the mixture contains at least 25% peat or CS.