Culture Substrate Research Articles

Evaluating the Feasibility of Hydrogel-Based Neural Cell Sprays.

Neurological injuries have poor prognoses with serious clinical sequelae. Stem cell transplantation enhances neural repair but is hampered by low graft survival (<ca. 5%), necessitating the development of approaches to enhance post-transplant cell viability. Intracerebral injection exerts high mechanical forces on transplant cells with risks of haemorrhage/infection. Transplant cell sprays can offer a non-invasive alternative. This study has assessed if the addition of protective, encapsulating polymer hydrogels to a cell spray format is feasible. Hydrogels (0.1% (1 mg/mL), 0.3% and 0.6% type I rat tail collagen) were trialled for spray deliverability. Cell-enriched hydrogels (containing mouse cortical astrocytes) were sprayed onto culture substrates. Astrocyte viability, cell-specific marker expression, morphology and proliferation were assessed at 24 h and 72 h post spraying. Intra-gel astrocytes and hydrogels could be co-stained using a double immunocytological technique (picrosirius red (PR)/DAB-peroxidase co-labelling). Astrocyte viability remained high post spraying with hydrogel encapsulation (>ca. 80%) and marker expression/proliferative potential of hydrogel-sprayed astrocytes was retained. Combining a cell spray format with polymer encapsulation technologies could form the basis of a non-invasive graft delivery method, offering potential advantages over current cell delivery approaches.

Investigation of Dielectric Measurement Model for Coconut Fiber Water Content and the Associated Factors

Coconut fiber has excellent permeability and water retention properties, making it a popular choice as an organic substrate for cultivation in the field of facility agriculture. To investigate the dielectric properties of coconut fiber and develop an accurate measurement model for dielectric water content, a vector network analyzer was used to measure the relative dielectric permittivity of coconut fiber and study the dielectric properties of coconut fiber under different volumetric water contents, temperatures, and salinities. The results indicated that the relative dielectric permittivity of the coconut fiber increased with increasing volumetric water content. The optimal frequency range for measuring the water content of the coconut fiber was 200 MHz–3 GHz, with an optimal frequency of 210 MHz. The dielectric measurement model for the water content in the coconut fiber established at this frequency demonstrated good applicability, with an R2 of 0.9704 and RMSE of 0.0453 cm3/cm3. At the same moisture level (θV of approximately 30% and 50%), the error in the real part of the dielectric permittivity was less than 1.35 between 15 and 35 °C, and the error in the moisture content was less than 0.0339 cm3/cm3. The error due to the salinity was 0.0750 cm3/cm3. The experimental results showed that the modeling of the water content measurement of coconut fiber at 210 MHz can be applied to general greenhouse environmental conditions. This study serves as a valuable reference for calibrating the moisture monitoring sensors used in greenhouses for coir cultivation substrates, emphasizing the need for non-destructive, real-time monitoring of moisture content.

Ulva rigida Valorization into Poly(3-hydroxybutyrate), Organic Acids and Functional Ingredients.

Halomonas elongata 1H9T is a moderate halophilic strain able to produce poly(3-hydroxybutyrate) (P(3HB)), a biodegradable plastic, and gluconic acid, a valuable organic acid with wide industrial applications. In this work, the green alga Ulva rigida was used as platform to produce cultivation substrates for microbial conversion as well as functional ingredients, targeting its full valorization. The liquor obtained by autohydrolysis presented the highest concentration of oligosaccharides and protein, being an interesting feedstock to produce functional ingredients. The acid and/or enzymatic hydrolysis liquors are adequate as substrates for microbial processes. Shake flask assays with H. elongata revealed that the N-rich liquor produced after acidic treatment was the best suited for cell growth while the N-poor liquor produced by the enzymatic treatment of acid-pretreated algae residues produced the highest P(3HB) titers of 4.4 g/L. These hydrolysates were used in fed-batch cultivations as carbon and protein sources for the co-production of gluconic acid and polymer achieving titers of 123.2 g/L and 7.2 g/L, respectively. Besides gluconic acid, the Krebs cycle intermediate 2-oxoglutaric acid, also called alpha-ketoglutaric acid (KGA), was produced. Therefore, the co-production of P(3HB) and acids may be of considerable interest as an algal biorefinery valorization strategy.

Bioactivity and Metabolomic Profile of Extracts Derived from Mycelial Solid Cultures of Hypsizygus marmoreus.

The beech mushroom (Hypsizygus marmoreus) is a highly nutritious, edible medicinal mushroom native to East Asia. The present research investigated the impact of different substrates on the metabolite compositions of H. marmoreus mycelia cultivated in vitro. The substrates tested included malt extract agar, malt extract agar enriched with barley malt, and malt extract agar enriched with grape pomace. The study also assessed antimicrobial and antiradical activities of the extracts against gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), gram-negative bacteria (Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa), yeasts (Candida albicans, C. tropicalis, and C. parapsilosis), and dermatophytes (Trichophyton mentagrophytes, T. tonsurans, T. rubrum, Arthroderma quadrifidum, A. gypseum, A. curreyi, and A. insingulare). The results revealed that the H. marmoreus mycelia extracts demonstrated antibacterial and antifungal activities against the tested microorganisms. Extracts obtained from the cultivation in substrates enriched with either barley malt or grape pomace exhibited the highest antibacterial activity among all the tested bacterial strains except for P. aeruginosa. The same extracts showed the highest inhibitory effect against C. albicans and C. parapsilosis. Noteworthy, the extract from the mushroom cultivated in the substrate enriched with grape pomace also exhibited remarkable efficacy against T. mentagrophytes and T. tonsurans. Terpenoid and carbapenem compounds could be related to the antimicrobial properties of the extracts from mushrooms cultivated in substrates enriched with grape pomace. In comparison, the higher antiradical properties could be related to the content of indole compounds. In conclusion, growth substrate selection affects the nutritional and medicinal properties of H. marmoreus, making it a valuable contribution to the understanding of the cultivation of this mushroom.

3D Printing of Bioactive Gel-like Double Emulsion into a Biocompatible Hierarchical Macroporous Self-Lubricating Scaffold for 3D Cell Culture.

The interconnected hierarchically porous structures are of key importance for potential applications as substrates for drug delivery, cell culture, and bioscaffolds, ensuring cell adhesion and sufficient diffusion of metabolites and nutrients. Here, encapsulation of a vitamin C-loaded gel-like double emulsion using a hydrophobic emulsifier and soy particles was performed to develop a bioactive bioink for 3D printing of highly porous scaffolds with enhanced cell biocompatibility. The produced double emulsions suggested a mechanical strength with the range of elastic moduli of soft tissues possessing a thixotropic feature and recoverable matrix. The outstanding flow behavior and viscoelasticity broaden the potential of gel-like double emulsion to engineer 3D scaffolds, in which 3D constructs showed a high level of porosity and excellent shape fidelity with antiwearing and self-lubricating properties. Investigation of cell viability and proliferation using fibroblasts (NIH-3T3) within vitamin C-loaded gel-like bioinks revealed that printed 3D scaffolds offered brilliant biocompatibility and cell adhesion. Compared to scaffolds without encapsulated vitamin C, 3D scaffolds containing vitamin C showed higher cell viability after 1 week of cell proliferation. This work represented a systematic investigation of hierarchical self-assembly in double emulsions and offered insights into mechanisms that control microstructure within supramolecular structures, which could be instructive for the design of advanced functional tissues.

Effects of substrate, water flow, and turbulence in raceway culture of the high-value agarophyte Pterocladiella capillacea

One reason most commercial seaweed aquaculture is done extensively is that land-based intensive culture is expensive. Seaweed growth in land-based raceways can be bolstered by high water flows and turbulence, and a better understanding of this may mitigate production costs. We investigated flow conditions in raceway culture of Pterocladiella capillacea, a source of pharmaceutical grade agar. We tested the effects of ploidy level (carpospores, tetraspores), substrate (rope, glass slide), and flow type in raceways (turbulent, laminar) and the laboratory (no flow) on the survival and growth of sporelings. We also assessed the effect of flow rates (1.5, 4, 10 cm s−1) and flow type (turbulent, laminar) on the growth of adult P. capillacea. Sporeling survival 48 h post-settlement was unaffected by flow type, and carpospores survived better than tetraspores on ropes. After four weeks, sporelings in raceways survived better on ropes than on glass slides, and sporelings derived from carpospores survived better on ropes than those from tetraspores. Sporelings grown in raceways were 5.8 times larger than those in the laboratory, but flow types had little effect within raceways. Seaweeds grown from carpospores grew 1.2 times larger than tetraspore in the raceways. Flow rate and type had little effect on the growth of adults, and their growth was slow possibly because of smothery by epiphytes. This study suggests P. capillacea spores are amenable to raceway culture, but adults do not fare as well in similar conditions.

Engineering dense tumor constructs via cellular contraction of extracellular matrix hydrogels.

Physical characteristics of solid tumors such as dense internal microarchitectures and pathological stiffness influence cancer progression and treatment. While it is routine to engineer culture substrates and scaffolds with elastic moduli that approximate tumors, these models often fail to capture characteristic internal microarchitectures such as densely compacted concentric ECM fibers at the stromal interface. Contractile mesenchymal cells can solve this engineering challenge by deforming, contracting, and compacting extracellular matrix (ECM) hydrogels to decrease tissue volume and increase tissue density. Here we demonstrate that allowing human fibroblasts of varying origins to freely contract collagen type I-containing hydrogels co-seeded with carcinoma cell spheroids produces a tissue engineered construct with structural features that mimic dense solid tumors in vivo. Morphometry and mechanical testing were conducted in tandem with biochemical analysis of proliferation and viability to confirm that dense carcinoma constructs engineered using this approach capture relevant physical characteristics of solid carcinomas in a tractable format that preserves viability and is amenable to extended culture. The reported method is adaptable to the use of multiple mesenchymal cell types and the inclusion of fibrin in the ECM combined with seeding of endothelial cells to produce prevascularized constructs. The physical dense carcinoma constructs engineered using this approach may provide more clinically relevant venues for studying cancer pathophysiology and the challenges associated with the delivery of macromolecular drugs and cellular immunotherapies to solid tumors.

Towards modular engineering of cell signalling: Topographically-textured microparticles induce osteogenesis via activation of canonical hedgehog signalling

Polymer microparticles possess great potential as functional building blocks for advanced bottom-up engineering of complex tissues. Tailoring the three-dimensional architectural features of culture substrates has been shown to induce osteogenesis in mesenchymal stem cells in vitro, but the molecular mechanisms underpinning this remain unclear. This study proposes a mechanism linking the activation of Hedgehog signalling to the osteoinductive effect of surface-engineered, topographically-textured polymeric microparticles. In this study, mesenchymal progenitor C3H10T1/2 cells were cultured on smooth and dimpled poly(D,l-lactide) microparticles to assess differences in viability, cellular morphology, proliferation, and expression of a range of Hedgehog signalling components and osteogenesis-related genes. Dimpled microparticles induced osteogenesis and activated the Hedgehog signalling pathway relative to smooth microparticles and 2D-cultured controls without the addition of exogenous biochemical factors. We observed upregulation of the osteogenesis markers Runt-related transcription factor2 (Runx2) and bone gamma-carboxyglutamate protein 2 (Bglap2), as well as the Hedgehog signalling components, glioma associated oncogene homolog 1 (Gli1), Patched1 (Ptch1), and Smoothened (Smo). Treatment with the Smo antagonist KAAD-cyclopamine confirmed the involvement of Smo in Gli1 target gene activation, with a significant reduction in the expression of Gli1, Runx2 and Bglap2 (p ≤ 0.001) following KAAD-cyclopamine treatment. Overall, our study demonstrates the role of the topographical microenvironment in the modulation of Hedgehog signalling, highlighting the potential for tailoring substrate topographical design to offer cell-instructive 3D microenvironments. Topographically-textured microparticles allow the modulation of Hedgehog signalling in vitro without adding exogenous biochemical agonists, thereby eliminating potential confounding artefacts in high-throughput drug screening applications.

Three-Dimensional Graphene Promotes the Proliferation of Cholinergic Neurons

Introduction: An early substantial loss of basal forebrain cholinergic neurons (BFCNs) is a common property of Alzheimer’s disease and the degeneration of functional BFCNs is related to learning and memory deficits. As a biocompatible and conductive scaffold for growth of neural stem cells, three-dimensional graphene foam (3D-GF) supports applications in tissue engineering and regenerative medicine. Although its effects on differentiation have been demonstrated, the effect of 3D-GF scaffold on the generation of BFCNs still remains unknown. Methods: In this study, we used 3D-GF as a culture substrate for neural progenitor cells (NPCs) and demonstrated that this scaffold material promotes the differentiation of BFCNs while maintaining excellent cell viability and proliferation. Results: Immunofluorescence analysis, real-time polymerase chain reaction, Western blotting, and ELISA revealed that the proportion of BFCNs at 21 days of differentiation reached approximately 30.5% on 3D-GF compared with TCPS group that only presented 9.7%. Furthermore, a cell adhesion study suggested that 3D-GF scaffold enhances the expression of adhesion proteins including vinculin, integrin, and N-cadherin. These findings indicate that 3D-GF scaffold materials are preferable candidates for the differentiation of BFCNs from NPCs. Conclusions: These results suggest new opportunities for the application of 3D-GF scaffold as a neural scaffold for cholinergic neurons therapies based on NPCs.

Implications of Cellular Mechanical Memory in Bioengineering.

The ability to maintain and differentiate cells in vitro is critical to many advances in the field of bioengineering. However, on traditional, stiff (E ≈ GPa) culture substrates, cells are subjected to sustained mechanical stress that can lead to phenotypic changes. Such changes may remain even after transferring the cells to another scaffold or engrafting them in vivo and bias the outcomes of the biological investigation or clinical treatment. This persistence─or mechanical memory─was initially observed for sustained myofibroblast activation of pulmonary fibroblasts after culturing them on stiff (E ≈ 100 kPa) substrates. Aspects of mechanical memory have now been described in many in vitro contexts. In this Review, we discuss the stiffness-induced effectors of mechanical memory: structural changes in the cytoskeleton and activity of transcription factors and epigenetic modifiers. We then focus on how mechanical memory impacts cell expansion and tissue regeneration outcomes in bioengineering applications relying on prolonged 2D plastic culture, such as stem cell therapies and disease models. We propose that alternatives to traditional cell culture substrates can be used to mitigate or erase mechanical memory and improve the efficiency of downstream cell-based bioengineering applications.

Impact of the Physical Cellular Microenvironment on the Structure and Function of a Model Hepatocyte Cell Line for Drug Toxicity Applications.

It is widely recognised that cells respond to their microenvironment, which has implications for cell culture practices. Growth cues provided by 2D cell culture substrates are far removed from native 3D tissue structure in vivo. Geometry is one of many factors that differs between in vitro culture and in vivo cellular environments. Cultured cells are far removed from their native counterparts and lose some of their predictive capability and reliability. In this study, we examine the cellular processes that occur when a cell is cultured on 2D or 3D surfaces for a short period of 8 days prior to its use in functional assays, which we term: "priming". We follow the process of mechanotransduction from cytoskeletal alterations, to changes to nuclear structure, leading to alterations in gene expression, protein expression and improved functional capabilities. In this study, we utilise HepG2 cells as a hepatocyte model cell line, due to their robustness for drug toxicity screening. Here, we demonstrate enhanced functionality and improved drug toxicity profiles that better reflect the in vivo clinical response. However, findings more broadly reflect in vitro cell culture practises across many areas of cell biology, demonstrating the fundamental impact of mechanotransduction in bioengineering and cell biology.

The role of a nanogrooved polydimethylsiloxane substrate on mesenchymal stem cells adhesion, self-renewing, and mechanical properties

Stem cells have a variety of applications in biological, reconstructive medicine, and tissue engineering studies owing to their capacity to differentiate into specific cell lineages and their self-renewal ability. Among the factors affecting stem cells, biochemical agents such as growth and differentiation factors, along with environmental cues including physical and mechanical characteristics of the cell microenvironment, are of interest in this field. Several studies have evaluated the impact of physical factors such as mechanical forces, electric or magnetic fields, and light pulses on stem cell behavior. Also, cell culture substrates that can simulate extracellular matrix in natural conditions are considered the most critical environmental factors. Hence, in this study, we investigated the effects of a specially designed polydimethylsiloxane substrate with a surface pattern for cell contact guidance on various biological and biophysical characteristics of mesenchymal stem cells. Plasma treatment of the substrate under lateral tension from 50 to 200 s caused the increased wavelength of the formed wrinkle structures from 500 ± 56 to 1052 ± 182 nm, respectively, leading to variations of stiffness affecting the cell behaviors. Moreover, by increasing the available surface and hydrophilicity of the substrate, the cell attachment quality improved by more than 46% compared to the control. Finally, results of the mechanics of mesenchymal stem cells on these substrates obtained by bio-AFM showed an increase in the stiffness of cells despite a slight decrease in Young's modulus parameter of different substrates. These changes in the intracellular mechanics can eventually stimulate the more pivotal mechanobiological effects on stem cell fate.

Direct deep UV lithography to micropattern PMMA for stem cell culture.

Microengineering is increasingly being used for controlling the microenvironment of stem cells. Here, a novel method for fabricating structures with subcellular dimensions in commonly available thermoplastic poly(methyl methacrylate) (PMMA) is shown. Microstructures are produced in PMMA substrates using Deep Ultraviolet lithography, and the effect of different developers is described. Microgrooves fabricated in PMMA are used for the neuronal differentiation of mouse embryonic stem cells (mESCs) directly on the polymer. The fabrication of 3D, curvilinear patterned surfaces is also highlighted. A 3D multilayered microfluidic chip is fabricated using this method, which includes a porous polycarbonate (PC) membrane as cell culture substrate. Besides directly manufacturing PMMA-based microfluidic devices, an application of the novel approach is shown where a reusable PMMA master is created for replicating microstructures with polydimethylsiloxane (PDMS). As an application example, microchannels fabricated in PDMS are used to selectively expose mESCs to soluble factors in a localized manner. The described microfabrication process offers a remarkably simple method to fabricate for example multifunctional topographical or microfluidic culture substrates outside cleanrooms, thereby using inexpensive and widely accessible equipment. The versatility of the underlying process could find various applications also in optical systems and surface modification of biomedical implants.

Sustainable Valorization of Organic Materials as Substrates for Soilless Crops in Protected Environments in the Venezuelan Andes

Agricultural production activities generate a large amount of waste, most of which is of organic origin from plant remains. These residues could easily be transformed into a resource, specifically, into a substrate for use in soilless cultivation; however, in most cases, they are not used. Therefore, a characterization of local agricultural residues was carried out to determine their use as a substrate for crop cultivation without soil in a protected environment. The selected substrates were Sphagnum peat, coir fiber, and compost, which were managed alone or in mixtures of 50/50, 75/25, or 25/75% v/v ratios. We also included a mixture of virgin mountain soil and earthworm humus (known as INIA mixture) because it is used by local growers. The results showed that the substrates based on coir fiber, peat, and mixtures of both presented suitable characteristics for horticultural crops. On the contrary, compost-based substrates had high pH values and low organic matter contents that could be improved before use. The relationship found between the evaluated parameters of each substrate allowed us to establish that variables, such as the content of organic matter and water at different tensions and particle sizes, can be utilized to make a quick selection of the substrates produced locally, which would lead to the use of waste in a way that is more consistent with sustainable agricultural production and minimal environmental impact, by being used in the production of crops in containers without soil. In addition, these results can be used as an alternative reference in localities where these residues are easily available.

Regolith simulant and green compost as substrates for plant cultivation in Mars space colonies

Regolith simulant and green compost as substrates for plant cultivation in Mars space colonies

The effect of AB mix fertilizer concentration on the yield of Marigold (Tagetes erecta) under hydroponic condition

In hydroponic substrate cultivation, nutrients are only sourced from fertilizers; therefore, fertilizer application must pay attention to concentration. The right concentration of fertilizer, namely AB Mix, affects the quality of marigold flowers (Tagetes erecta). The purpose of this study was to examine the effect of flower AB mix fertilizer concentration and obtain the best AB mix fertilizer concentration for marigold flower yields on cultivated hydroponic substrate. This research was carried out in May–July 2022 at the screen house of the Horticulture Industry Technology Study Program. The study used Group Randomized Design (GRD), with AB mix fertilizer treatment tested at a concentration of 1000; 1100; 1200; 1300; 1400; 1500; and 1600 ppm. The data obtained were analyzed with various fingerprints and continued with the honest significant difference (HSD) test level of 5% if there was a real effect. The parameters observed are the number of flowers, the diameter of the flower, and the fresh weight of the flower per plant. The results obtained are: 1. The concentration of AB mix fertilizer has a significant influence on the number of flowers, flower diameter, and fresh weight of flowers per plant). AB mix fertilizer with a concentration of 1,200 ppm is the best concentration because it produces marigold plants with the highest number of flowers (17.67 florets), the widest flower diameter (in the first harvest it was 79.31 mm, the second harvest was 78.12 mm, and the third harvest was 78.16 mm), and the heaviest flower weight (in the first harvest it was 15.11 g, the second harvest was 14.33 g, and the third harvest was 12.67 g).

Chicken feather waste degradation by Malbranchea cinnamomea and its application on plant growth and metabolites of Vicia faba plant

Keratinase are proteolytic enzymes that have received a lot of interest for their ability to transform the keratinous wastes that are a major source of environmental pollution. The most effective isolate that produced more keratinase was Malbranchea cinnamomea CBS 343.55. The enzyme activity was higher at 37 °C, pH 5.6; the incubation period was 7 days; and 1.0 ml was found to be the best inoculum size for keratinase production. Under solid substrate cultivation, the best substrate loadage for maximal keratinase production was 3.0 g/500 ml conical flask for M. cinnamomea CBS 343.55. The purified keratinase was stable between pH 3 and 8 and stable at temperatures ranging from 40 to 90 °C, with optimal activity at 45 °C and pH 8. The relative activity of keratinase was raised in the presence of the ions Na+, Mg2+, Mn2+, Ca2+, Co2+, Fe2+, Fe3+, and mercaptoethanol, but decreased in the presence of Hg2+, Zn2+, Cu2+, Ag2+, pb2+, ethylenediaminetetraacetic acid, and phenylmethylsulfonyfluoride. Chicken feather hydrolysate of M. cinnamomea produced total amino acids (329.0 mg/100 mL). In soil treated with feather compost and a fungus strain, there was a rise in the level of N, P, and K. However, the feather compost had a favorable impact on the growth, relative water content, photosynthetic pigments, antioxidant activity, total phenol and proline content of Vicia faba plant. In conclusion, the present study revealed that M. cinnamomea CBS 343.55O has potential applications in the biodegradation of chicken feathers and the value-addition of poultry waste as a biofertilizer.

Cell culture design for homogeneous proliferation of cells in three-dimensional nonwoven polymer scaffolds

The objective of this study is to establish strategies to uniformly proliferate cells in a three-dimensional nonwoven polyethylene terephthalate (PET)/ethylene vinyl alcohol (EVOH) scaffold by simple adjustments in seeding and culture methods and the scaffold design. The combined dynamic and static seeding (intermittent agitations at 300 rpm with 1 h interval) resulted in the highest seeding efficiency (71%) comparing to the static and continuous agitating seeding methods. Cell-attached scaffolds were cultivated under different conditions. The stirring culture permitted cells to proliferate to a significantly greater extent than the static or agitating cultures, although faster cell proliferation in the outer region of the scaffold was observed. Next, based on this observation, scaffolds were opened with holes to alleviate the cell aggregation. The effect of hole size and number of scaffolds on the distribution of cells proliferated in the scaffold was evaluated. Two of 1-mm holes showed to be an optimal adjustment to allow cells to proliferate in a homogeneous manner. After 14 days culture, both of the holes were filled by cells proliferated with a fourfold increase in the cell number. The cell viability in the scaffolds was also high upon evaluating the live/dead and 3[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) staining examinations. Different cell types of 3T3-L1, C3H/10T1/2, and KUM6 cells showed similar behavior of cell proliferation and distribution in the scaffold, indicating the applicability of the established procedure. It is concluded that the nonwoven PET/EVOH scaffold serves as a potential cell culture substrate for an efficient cell proliferation.

The Impact of the Growing Substrate on Morphological and Biochemical Features of Salicornia europaea L.

Nowadays, intensive agriculture correlated with the impact of climate change has led to nutrient soil depletion and the salinization of agricultural lands, making them unsuitable for conventional agricultural crops, with a direct impact on the food industry. Therefore, it is necessary to find sustainable alternative solutions that satisfy the needs of both consumers and food production. One such solution may be represented by salt-tolerant species that can fulfill food requirements. One of the most promising salt-tolerant plant species that can be used is Salicornia europaea L. The present work was conducted in greenhouse conditions, and the adaptability of the species on different cultivation substrates was investigated by means of monitoring the plant indicators such as cuticle, epidermis, parenchyma, polyphenols content, and minerals. Moreover, the correlation between the polyphenol and mineral contents was highlighted. Therefore, three cultivation substrates with different levels of salinity/electrical conductivity were used. The reference (I) for biochemical indicators was represented by the plant grown in natural salinity conditions in the Southeast region of Romania. The results indicate that Salicornia europaea L. can be grown on different cultivation substrates other than salted soils, the plant showing the capacity to accumulate bioactive compounds similar to natively grown ones.

Turn Waste into Treasure: Spent Substrates of Auricularia heimuer Can Be Used as the Substrate for Lepista sordida Cultivation

Mudanjiang is a major producer of black wood ear (Auricularia heimuer) mushrooms in China. It has been estimated that more than 1.5 million tons of spent substrates of A. heimuer (SSA) are produced each year. Most of these are discarded or burned and have become an important source of pollution, which urgently merits research to find appropriate uses for them. To explore the feasibility of SSA as a substrate for cultivating Lepista sordida mushrooms, experiments were conducted to assess the effects of the addition of 0, 40%, 73%, and 98% SSA on the days required for the mycelia to fully colonize the substrate and initiate primordia, biological efficiency (BE), yield, and composition of the chemical biomass of the L. sordida fruiting bodies. The yield of fruiting bodies with 0, 40%, 73%, and 98% SSA supplementation for three flushes was 3.90 ± 0.74 kg m−2, 4.06 ± 0.77 kg m−2, 4.03 ± 0.62 kg m−2, and 4.51 ± 0.65 kg m−2, respectively. The addition of 98% SSA significantly delayed the number of days for the mycelia to fully colonize and form primordia by 6 and 3 d, respectively. This treatment also significantly increased the yield and BE by 15.64% compared with that of the control group. The crude polysaccharide content of 25.64 ± 0.38 g 100 g−1 was higher in the samples grown on the 98% SSA substrate, which was shown to increase by 78.93% compared with that of the control substrate with 73% corn straw (14.33 ± 0.03 g 100 g−1). The content of crude protein of 51.10 ± 0.08 g 100 g−1 was higher in the samples grown on the 40% SSA substrate, which increased by 11.14% compared with the protein content of the control group. This study reveals that SSA would be a good substrate for the cultivation of L. sordida and is an efficient, promising, and cost-effective substrate additive that can improve the quality and yield of these mushrooms, while substantially reducing the problems of disposing of SSA.