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Spent mushroom substrate (SMS) is the residual biomass generated after harvesting the fruitbodies of edible/medicinal fungi. Disposal of SMS, the main by-product of the mushroom cultivation process, often leads to serious environmental problems and is financially demanding. Efficient recycling and valorization of SMS are crucial for the sustainable development of the mushroom industry in the frame of the circular economy principles. The physical properties and chemical composition of SMS are a solid fundament for developing several applications, and recent literature shows an increasing research interest in exploiting that inherent potential. This review provides a thorough outlook on SMS exploitation possibilities and discusses critically recent findings related to specific applications in plant and mushroom cultivation, animal husbandry, and recovery of enzymes and bioactive compounds.

The extraction of bioactive compounds and cellulose saccharification are potential directions for the valorization of spent mushroom substrate (SMS). Therefore, investigating the suitability of different extraction methods for recovering bioactive compounds from SMS and how the extraction affects the enzymatic saccharification is of uppermost relevance. In this work, bioactive compounds were extracted from Pleurotus spp. SMS using four extraction methods. For Soxhlet extraction (SoE), a 40:60 ethanol/water mixture gave the highest extraction efficiency (EE) (69.9-71.1%) among the seven solvent systems assayed. Reflux extraction with 40:60 ethanol/water increased the extraction yield and EE compared to SoE. A shorter reflux time yielded a higher extraction of carbohydrates than SoE, while a longer time was more effective for extracting phenolics. The extracts from 240 min of reflux had comparable antioxidant activity (0.3-0.5 mM GAE) with that achieved for SoE. Ultrasound-assisted extraction (UAE) at 65 °C for 60 min allowed an EE (~82%) higher than that achieved by either reflux for up to 150 min or SoE. Subcritical water extraction (SWE) at 150 °C resulted in the best extraction parameters among all the tested methods. Vanillic acid and chlorogenic acid were the primary phenolic acids identified in the extracts. A good correlation between the concentration of caffeic acid and the antioxidant activity of the extracts was found. Saccharification tests revealed an enhancement of the enzymatic digestibility of SMS cellulose after the extraction of bioactive compounds. The findings of this initial study provide indications on new research directions for maximizing the recovery of bioactive compounds and fermentable sugars from SMS.

Background: Rapid diagnostics of pathogens is essential to prescribe appropriate antibiotic therapy. The current methods for pathogen detection require the bacteria to grow in a culture medium, which is time-consuming. This increases the mortality rate and global burden of antimicrobial resistance. Culture-free detection methods are still under development and are not common in the clinical routine. Therefore, decreasing the culture time for accurately detecting infection and resistance is vital for diagnosis. Methods: This study investigated easy-to-implement factors (in a minimal laboratory set-up), including inoculum size, incubation temperature, and additional supplementation ( e.g. , vitamin B12 and trace metals), that can significantly reduce the bacterial lag time (t lag ). These factors were arranged in simple two-level factorial designs using Gram-positive cocci ( Staphylococcus aureus ), Gram-positive bacilli ( Bacillus subtilis ), and Gram-negative bacilli ( Escherichia coli and Pseudomonas aeruginosa ) bacteria, including clinical isolates with known antimicrobial resistance profiles. Blood samples spiked with a clinical isolate of E. coli CCUG 17620 (Culture Collection University of Gothenburg) were also tested to see the effect of elevated incubation temperature on bacterial growth in blood cultures. Results: We observed that increased incubation temperature (42°C) along with vitamin B12 supplementation significantly reduced the t lag (10 - 115 minutes or 4% - 49%) in pure clinical isolates and blood samples spiked with E. coli CCUG17620. In the case of the blood sample, PCR results also detected bacterial DNA after only 3h of incubation and at three times the CFU/mL. Conclusion: Enrichment of bacterial culture media with growth supplements such as vitamin B12 and increased incubation temperature can be a cheap and rapid method for the early detection of pathogens. This proof-of-concept study is restricted to a few bacterial strains and growth conditions. In the future, the effect of other growth conditions and difficult-to-culture bacteria should be explored to shorten the lag phase., Competing Interests: No competing interests were disclosed., (Copyright: © 2023 Ali J et al.)

Background: The rapid diagnostics of pathogens is essential to prescribe appropriate and early antibiotic therapy. The current methods for pathogen detection require the bacteria to grow in a culture medium, which is time-consuming. This increases the mortality rate and the global burden of antimicrobial resistance. Culture-free detection methods are still under development and are not used in the clinical routine. Therefore decreasing the culture time for accurate detection of infection and resistance is vital for diagnosis. Methods: In this study, we wanted to investigate easy-to-implement factors (in a minimal laboratory set-up), including inoculum size, incubation temperature, and additional supplementation ( e.g. , vitamin B12 and trace metals), that can significantly reduce the lag time (t lag ). These factors were arranged in simple two-level factorial designs using Gram-positive ( Escherichia coli and Pseudomonas aeruginosa ) and Gram-negative ( Staphylococcus aureus and Bacillus subtilis ) bacteria, including clinical isolates with known antimicrobial resistance profiles. Blood samples spiked with a clinical isolate of E. coli CCUG17620 were also tested to see the effect of elevated incubation temperature on bacterial growth in blood cultures. Results: We observed that increased incubation temperature (42°C) along with vitamin B12 supplementation significantly reduced the t lag (10 - 115 minutes or 4% - 49%) in pure clinical isolates and blood samples spiked with E. coli CCUG17620. In the case of the blood sample, PCR results also detected bacterial DNA after only 3h of incubation and at three times the CFU/mL. Conclusions: Enrichment of bacterial culture media with growth supplements such as vitamin B12 and increased incubation temperature can be a cheap and rapid method for the early detection of pathogens. This is a proof-of-concept study restricted to a few bacterial strains and growth conditions. In the future, the effect of other growth conditions and difficult-to-culture bacteria should be explored to shorten the lag phase., Competing Interests: No competing interests were disclosed., (Copyright: © 2023 Ali J et al.)

Complement component 5 (C5) is an essential factor of the defensive complement system in all vertebrates. We report the characterization of C5 cDNA and protein from Atlantic salmon (Salmo salar), a teleost fish species of high importance in aquaculture. The C5 cDNA cloned from liver is 5079 nucleotides long, whose translation product has a molecular weight of 190 kDa, with the classical β-α orientation and motifs/sites for β-α cleavage ( 678 RPKR 681 ) and cleavage by C5 convertases (R 758 ). Mass spectrometric analysis show a single N-linked, biantennary, complex glycan at N 1125 . Moreover, the N-linked glycan displays an unusual modification in the form of acetylated sialic acid residues. Three anti-C5 antisera produced in mice using purified C5 worked in immunohistochemical analyses of formalin fixed liver tissue. The purification method, whereby inactive and activated (C5b) forms were isolated, opens for interesting studies on the complement function in fish, including possible connection to stress, disease and glycosylation., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Phenolic acids including chlorogenic acids are major polyphenolic compounds found in Jerusalem artichoke ( Helianthus tuberosus L.). The plant itself is an emerging biorefinery crop due to the inulin-rich tubers, a bioethanol feedstock, but the aerial parts represent a rich source of bioactive compounds. We have determined the level of major phenolic acids in extracts of four plant organs: tuber, leaf, flower, and stem. Employing three heating conditions (20 °C, 60 °C, and microwaving), corrected total phenolic content (TPC) was highest in the leaves (4.5-5.7 mg gallic acid equivalents g -1 dry substance), followed by flower (2.1-2.9), tuber (0.9-1.4), and lowest in stem extracts (0.1-0.2). A previously overlooked interference of the Folin-Ciocalteu assay, namely a signal contribution from ascorbic acid, caused overestimation of TPC in various organs ranging from 65% to 94%. Radical scavenging activity of extracts correlated significantly with TPC, both on corrected (R 2 = 0.841) and uncorrected (R 2 = 0.884) values. Out of the identified phenolic acids determined by quantitative HPLC-UV analysis, chlorogenic and dicaffeoylquinic acids accounted for 72-82% of corrected TPC in leaf and tuber extracts. Optimization of leaf extraction was tested in a 2 3 -factorial Central Composite Face (CCF) design. Temperature was the most important model term, and a solvent strength of less than 50% ethanol promoted the highest TPC yields. Further developments in extraction processing of crop residues may open avenues for improving the utilization of Jerusalem artichoke in valuable products.

Background: The interplay between diet, gut bacteria and health still remain enigmatic. Here, we addressed this issue through the investigation of the effect of crystalline cellulose on the earthworm Lumbricus terrestris gut microbiota composition and survival., Methods: Earthworm gut contents were analyzed after 14 days of feeding using a mixed 16S rRNA gene sequencing approach, in addition to direct measurements of cellulase activity. The survival of earthworms was followed each week for 17 weeks., Results: We found a tendency that the crystalline cellulose fed earthworms survived better than the high energy fed earthworms (p=0.08). Independent of feeding we found that the bacterial group related to Ferrimonadaceae was correlated to an increased lifespan (p=0.01). We also found a positive correlation between Ruminococcaceae related bacteria and cellulase activity in the earthworm gut (p=0.05). Surprisingly, however, the cellulase activity was not correlated to the feeding regime., Conclusion: Taken together, the interactions between diet, gut microbiota and lifespan seem complex.

The dynamics of all ecosystems are dictated by intrinsic, density-dependent mechanisms and by density-independent environmental forcing. In spite of the importance of the gastrointestinal microbiota in health and disease, the ecology of this system remains largely unknown. Here, we take an ecological approach to gut microbial community analysis, with statistical modelling of time series data from chemostats. This approach removes effects of host forcing, allowing us to describe a network of intrinsic interactions determining the dynamic structure of an experimental gut microbiota. Surprisingly, the main colonization pattern in this simplified model system resembled that of the human infant gut, suggesting a potentially important role of density-dependent interactions in the early gut microbiota. Knowledge of ecological structures in microbial systems may provide us with a means of controlling such systems by modifying the strength and nature of interactions among microbes and between the microbes and their environment., (© 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.)

An alkaline deoxyribonuclease (DNase) from cod pancreatic tissue has been characterized. The enzyme is a DNase I type endonuclease and hydrolyzes effectively both native and denatured DNA. Monomeric actin inhibits the enzyme reaction. The enzyme obeys Michaelis-Menten kinetics and the apparent Km value for native linear duplex DNA is 33 µg/ml. The cod DNase opens supercoiled plasmid DNA, by introducing adjacent nicks in both strands, possibly separated by 5-10 nucleotides. DNA hydrolyzed by cod DNase functions as substrates both for DNA polymerase and ligase, and the nicks therefore contain 5'-phosphoryl and 3'-hydroxyl groups. Optimum concentrations of divalent cations are 5 mM Mg(2+), 0.63 mM Mn(2+) and 0.075 mM Ca(2+). However, Ca(2+) is apparently not essential for the enzymatic functions. The enzyme has a narrow temperature optimum at 42°C and is thermolabile above 50°C; however, Mn(2+) shifts the optimum slightly to 45°C by causing increased temperature stability. The cod DNase reaction is inhibited by the DNA intercalating compounds actinomycin D and ethidium bromide. Histidine-modifying reagents such as tosyl phenylalanyl chloromethylketone and diethyl pyrocarbonate inhibit the enzyme activity, but the cod DNase is insensitive to disulfide-reducing agents.

The key step in the purification of a deoxyribonuclease (DNase) from extracts of cod (Gadus morhua L.) pyloric caeca, is the selective retention of the enzyme by anion exchange chromatography. The cod DNase purification on Q-Sepharose Fast Flow (Pharmacia) was optimized, using a 60 ml fixed-bed column. In combination with titration curve analysis, we have screened the effect of buffer pHs, feed conductivity and protein loading, on the product recovery and purity. We have developed elution conditions which allow effective separation of the cod DNase from bounded impurities, such as proteinases and nucleic acids. Low levels of these impurities were regarded as essential for the desired product quality. The optimum resolution and maximum purification (ca. 20-fold increase in specific activity) of DNase, was, however, achieved at low protein loading (2.6 mg ml-1 gel), corresponding to less than 4% of the dynamic bed capacity. Scale-up to a 2.5 l pilot scale column (axial flow) and a 0.25 l radial flow column showed that the separation and yield obtained at laboratory scale was retained, and was independent of column geometry and bed height. The implications for a production scale scenario of 100 g of fractionated protein, are also discussed, as well as process hygiene. The optimization described herein adds further knowledge to the treatment of fish waste and the downstream processing of valuable biochemicals from marine raw material.

A deoxyribonuclease (DNase) of pancreatic origin has been purified from extracts of the pyloric caeca from Atlantic cod (Gadus morhua L.). The crude extract was prepared by mincing frozen caeca tissue in equal volumes of buffer. The enzyme was isolated from the supernatant after streptomycin sulfate precipitation and centrifugation. The purification scheme further included chromatography on Q-Sepharose Fast Flow and hydroxyapatite columns. Affinity adsorption chromatography of the hydroxyapatite fraction on 8-(6-aminohexyl)-amino-5'-AMP-Sepharose, revealed an apparently homogeneous protein with molecular weight of 35,000 Da as judged by NaDodSO4-PAGE. In sum a 644-fold enzymatic enrichment and 3.5% total enzyme recovery was achieved. The cod enzyme resembles DNase I-type enzymes with an alkaline pH activity optimum and shows dependency for Mg(2+). The pI of the enzyme is 6.5 as determined by isoelectric focusing and DNase-zymography. Our findings suggest that the nuclease is a member of the cod's digestive enzymes secreted from the connective tissue surrounding the caeca.

Findings in this paper show a strong correlation between subjects' age and their olfactory sensitivity to the "fishy" odor of trimethylamine, with youngest subjects being most sensitive and adult subjects least sensitive to this odor. This was due to a high percentage of highly sensitive subjects in the youngest age groups; this percentage decreased with age. Data further support the notion that trimethylamine sensitivity is independent of sex. The sensitivity to trimethylamine per se showed no significant covariations with the subjects' preferences for or aversions against fish as food and is probably of minor importance for fish food acceptability.

A set-up for casting fluorescent indicator agarose gels on ultrathin polyacrylamide microelectrophoresis gel media (Pharmacia PhastGel media) is described. The zymogram system allows a rapid and sensitive detection of deoxyribonuclease in various gel media, following isoelectric focusing, native and sodium dodecyl sulfate-polyacrylamide gel electrophoresis.