Enhancing Biomass and ß-Glucan Yield from Oyster Mushroom Pleurotus ostreatus (Agaricomycetes) Mycelia through Extract Valorization.

This study aimed to examine the impact of mushroom extract-based solid media on the ß-glucan content, growth rate, density, and biomass content of Pleurotus ostreatus (oyster mushroom) mycelia. Fresh, high-quality raw P. ostreatus were washed, sliced, and heated in a sealed pressure cooker at 90°C for 4 h in the drying cabinet. Following the heating process, centrifugation was carried out. Different concentrations of Pleurotus ostreatus extract were mixed with distilled water (0%, 25%, 50%, 75%, and 100%) and prepared for a sterile solid media. A malt extract-based medium was maintained as a control. This study focuses on the growth performance of P. ostreatus mycelium on its own mushroom extract-based culture medium which holds considerable economic and environmental significance. During the six-day observation period, the mycelium exhibited consistent growth across all tested media, maintaining a steady growth rate of 15 mm. The increased content of mushroom extract resulted from the enhanced density of the mycelia and biomass content. It can be inferred that when media containing less than 25% of mushroom extract dilution is used, ß-glucan can be formed in smaller amounts. Further research is needed to explore mushroom extract derived from different types of mushroom byproducts, which fail to meet commercial standards.

Size Matters: Influence of Particle Size on Antioxidant, ß-Glucan, and Anti-Inflammatory Potential in Pleurotus floridanus (Agaricomycetes).

Cellular damage resulting from elevated levels of free radicals can lead to persistent health issues. Pleurotus floridanus, an edible white oyster mushroom, is rich in ß-glucans with potent antioxidant and anti-inflammatory properties. In this research, we examined the ß-glucan content, total phenolic content, as well as antioxidant and anti-inflammatory potential of hot water extracts with varying particle sizes (< 75, 75-154, 154-300, and 300-600 µm) of both whole and sliced fruiting bodies of P. floridanus. The findings revealed that the в-glucan content increased as the particle size increased, although no significant differences were observed. Conversely, smaller particle sizes (< 75 µm) of whole and sliced fruiting bodies of P. floridanus exhibited higher phenolic content, 2,2-diphenyl-1-picryl-hy-drazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, and reducing ability compared with larger particle size (> 75 µm). Of the four samples (AW2, AW3, AS1, and AS2) with the highest antioxidant activity selected for anti-inflammatory assays, all demonstrated the ability to reduce nitric oxide and tumor necrosis factor-alpha levels, but did not enhance interleukin-10 expression in lipopolysaccharide-stimulated RAW264.7 cells. Interestingly, particle size < 75 to 300 µm did not appear to influence the anti-inflammatory activity, because no significant differences were observed among the particle sizes. Therefore, a particle size < 300 µm in a P. floridanus hot water extract could serve as a valuable source of antioxidant and anti-inflammatory compounds to counteract the harmful effects of free radicals.

Quantitative trait loci analysis for molecular markers linked to agricultural traits of Pleurotus ostreatus.

Pleurotus ostreatus is a global mushroom crop with nutritional and medicinal benefits. However, the genetic basis of several commercial traits remains unknown. To address this, we analyzed the quantitative trait loci (QTLs) for two representative cultivars, "Heuktari" and "Miso," with apparently distinct alleles. A genetic map with 11 linkage groups was constructed, in which 27 QTLs were assigned to 14 traits. The explained phenotypic variations in QTLs ranged from 7.8% to 22.0%. Relatively high LOD values of 6.190 and 5.485 were estimated for the pinheading period and the number of valid stipes, respectively. Some QTL-derived molecular markers showed potential enhancement rates of selection precision in inbred lines, especially for cap shape (50%) and cap thickness (30%). Candidate genes were inferred from the QTL regions and validated using qRT-PCR, particularly for the cysteine and glutathione pathway, in relation to cap yellowness. The molecular markers in this study are expected to facilitate the breeding of the Heuktari and Miso lines and provide probes to identify related genes in P. ostreatus.

An ecofriendly approach to bioremediate nickel oxide nanoparticles using a macrofungus, Pleurotus fossulatus.

Nowadays, nickel oxide nanoparticles are in great demands owing to their use in many sectors. These nanoparticles may release into aquatic environment from different industries and cause negative effect on aquatic flora and fauna. Therefore, an effective and efficient method is required to remove these nanoparticles from contaminated water. Hence, the aim of this study was to bioremediate nickel oxide nanoparticles using a macrofungus, Pleurotus fossulatus, and to analyze its impact on fungal physiology. For this purpose, fungal spawns were inoculated in malt dextrose agar media containing different concentrations of nickel oxide nanoparticles (24 mg/l, 48 mg/l, and 100 mg/l) as well as control group (having no nickel oxide nanoparticles) and allowed to grow for a period of 20 days. Fungal mycelia as well as media were collected at different time intervals (5th day, 10th day, 15th day, and 20th day) for evaluation of Ni concentration and different biochemical parameters. Ni removal efficiency of P. fossulatus from media was found to be highest in 48 mg/l (66.98%) followed by 24 mg/l (60.83%) and 100 mg/l (18.03%), respectively. Increased level of metallothionein, lipid peroxidation, activity of different antioxidant enzymes (superoxide dismutase, catalase, glutathione s transferase, glutathione reductase), activity of ligninolytic enzymes (laccase, lignin peroxidase, manganese peroxidase), and shift in FTIR spectra were also reported in mycelia cultured in malt dextrose agar media containing nickel oxide nanoparticles. This study suggests that P. fossulatus has great efficiency to remediate nanoparticles from contaminated water and it can be utilized as potential agent in wastewater treatment plants by different industries.

Crystal Structure of the Native Chromoprotein from Pleurotus salmoneostramineus Provides Insights into the Pigmentation Mechanism.

The pink-colored protein from the fungus Pleurotus salmoneostramineus (PsPCP) possesses unusual primary sequences with little resemblance to those of known proteins and exhibits a red color in aqueous solution. To understand the pigmentation mechanism of PsPCP, we elucidated the X-ray crystal structure of the native PsPCP. We identified a highly conjugated polyene ligand 2-dehydro-3-deoxylaetiporic acid A as a chromophore ligand, whose solution exhibits yellow. The crystal structure of PsPCP indicated that the ligand is secured in the central cavity and anchored at both termini by hydrophilic interactions and that surrounding residues show CH-pi and C-H···O hydrogen bondings. Geometrical analyses of the bound ligand demonstrated that the conjugated C-C and C═C bonds exhibit similar bond distances. The result indicated enhanced electron delocalization within the conjugated CC bond system, resulting in a redshift of the chromophore ligand. The computational estimates of the UV-vis spectra support the view that the electron delocalization within the conjugated CC bonds system of the bound ligand, induced by the specific ligand geometry within a limited space of PsPCP cavity, is responsible for the red pigmentation of PsPCP. Thus, we propose that the coloring mechanism of PsPCP, which constrains the geometry of a highly conjugated polyene ligand, is a novel type of pigment chemistry.

Utilization of local agro-industrial by-products based substrates to enhance production and dietary value of mushroom (P. ostreatus) in Ethiopia.

Food insecurity and malnutrition are serious problems in many developing countries, including Ethiopia. This situation warrants an urgent need for the diversification of food sources with enhanced productivity. This study was aimed at contributing to the food security in Ethiopia through cultivation of Pleurotus ostreatus mushrooms using sustainable and locally available agro-industrial byproduct-based substrates in parallel with pollution control. Ten substrates were prepared using sugarcane bagasse, filter cake, trash, cotton seed hull and animal waste, namely cow dung and horse and chicken manure. The effect of each substrate (treatment) on the yields, biological efficiency, nutritional composition, and mineral contents of Pleurotus ostreatus mushroom species was evaluated at the Ethiopian Forest Products Innovation Center, Addis Ababa, Ethiopia. The results obtained indicate that a significantly higher (p < 0.05) yield and biological efficiency were recorded from the mushroom cultivated on S2 substrate containing a mixture of 80% sugarcane bagasse, 12% cow dung, and 8% cotton seed hull. Moreover, substrate containing sugarcane bagasse mixed with cotton seed hull, cow dung, and chicken manure significantly (p < 0.05) increased the yields and biological efficiency of the mushroom. The content of protein, crude fat, fiber, and carbohydrates of the mushroom cultivated from all the utilized substrates were in the range of 17.30-21.5, 1.77-2.52, 31.03-34.38, and 28.02-39.74%, respectively. The critical macro-elements are abundant in the mushroom in the order of potassium, magnesium, calcium, and sodium. The mushrooms cultivated on all the substrates were rich in essential micro-elements in the order of iron and zinc. It was found that substrate preparation and formulation significantly (p < 0.05) improved the yields, biological efficiency, nutritive values, and mineral contents of the mushroom. The use of these by-products as substrates is sustainable and environmentally friendly and allows the production of mushroom with high nutritional value on a sustainable basis in order to enhance food security in the country.

Optimization of polysaccharide conditions and analysis of antioxidant capacity in the co-culture of Sanghuangporus vaninii and Pleurotus sapidus.

Fungal polysaccharides are commonly utilized in the food industry and biomedical fields as a natural and safe immune modulator. Co-culturing is a valuable method for enhancing the production of secondary metabolites. This study used intracellular polysaccharide (IPS) content as a screening index, co-culturing seven different fungi with Sanghuangporus vaninii. The seed pre-culture liquid culture time was selected through screening, and conditions were assessed using single factor experimentation, a Plackett-Burman (PB) design, and response surface methodology (RSM) optimization. RSM optimization was conducted, leading to the measurement of antioxidant capacity. Results indicated that the co-culture of S. vaninii and Pleurotus sapidus exhibited the most effective outcome. Specifically, pre-culturing S. vaninii and P. sapidus seed cultures for 2 days and 0 days, respectively, followed by co-culturing, significantly increased IPS content compared to single-strain culturing. Further optimization of co-culture conditions revealed that yeast extract concentration, liquid volume, and S. vaninii inoculum ratio notably influenced IPS content in the order of yeast extract concentration > liquid volume > S. vaninii inoculum ratio. Under the optimal conditions, IPS content reached 69.9626 mg/g, a 17.04% increase from pre-optimization co-culture conditions. Antioxidant capacity testing demonstrated that co-cultured IPS exhibited greater scavenging abilities for DPPH and ABTS free radicals compared to single strain cultures. These findings highlight the potential of co-culturing S. vaninii and P. sapidus to enhance IPS content and improve antioxidant capacity, presenting an effective strategy for increasing fungal polysaccharide production.

Accumulation of Bisphenol A® by Pleurotus spp.-Flow Injection Analysis.

A specific feature of mushrooms (including those of the genus Pleurotus) is their natural ability to absorb and accumulate many chemical substances present in their immediate environment, which makes them an excellent natural sorption material. Hence, fruiting bodies of mushrooms have been recognized for years as excellent indicators of the environment, reflecting its current state. Nevertheless, mushrooms can accumulate both health-promoting substances, such as bioelements, and toxic substances, such as heavy metals and organic compounds, including bisphenol A® (BPA). This organic chemical compound in the phenol group, although it has been withdrawn in the EU since 2010, is widely present in the environment around us. In the present experiment, we aimed to determine the effect of adding BPA to liquid media for in vitro cultures of Pleurotus spp. The biomass increases were determined. Moreover, the degrees of adsorption and desorption of BPA from the obtained freeze-dried biomass in two different environments (neutral and acidic) were determined as a function of time. This is the first study to determine the bioavailability of adsorbed BPA in obtained biomass by extracting the mycelium into artificial digestive juices in a model digestive system. BPA was added to the liquid Oddoux medium in the following amounts: 0.01, 0.5, and 0.5 g/250 mL of medium. The amounts of adsorbed and desorbed BPA were determined by flow injection analysis (FIA) with amperometric detection. The addition of BPA to the substrate reduced the biomass growth in each of the discussed cases. BPA adsorption by the mycelium occurred at over 90% and depended on the morphology of the mushroom (structure, surface development, and pore size). BPA desorption depended on the pH of the environment and the desorption time. Mushrooms are an excellent natural remedial material, but BPA is extracted into artificial digestive juices; therefore, consuming mushrooms from industrialized areas may have health consequences for our bodies.

Transmission of Oyster Mushroom Spherical Virus to Progeny via Basidiospores and Horizontally to a New Host Pleurotus floridanus.

Mycoviruses are usually transmitted horizontally via hyphal anastomosis and vertically through sporulation in natural settings. Oyster mushroom spherical virus (OMSV) is a mycovirus that infects Pleurotus ostreatus, with horizontal transmission via hyphal anastomosis. However, whether OMSV can be vertically transmitted is unclear. This study aimed to investigate the transmission characteristics of OMSV to progeny via basidiospores and horizontally to a new host. A total of 37 single-basidiospore offspring were obtained from OMSV-infected P. ostreatus and Pleurotus pulmonarius for Western blot detection of OMSV. The OMSV-carrying rate among monokaryotic isolates was 19% in P. ostreatus and 44% in P. pulmonarius. Then, OMSV-free and OMSV-infected monokaryotic isolates were selected for hybridization with harvested dikaryotic progeny strains. Western blot analyses of the offspring revealed that the OMSV transmission efficiency was 50% in P. ostreatus and 75% in P. pulmonarius, indicating vertical transmission via sexual basidiospores. Furthermore, we observed the horizontal transfer of OMSV from P. pulmonarius to Pleurotus floridanus. OMSV infection in P. floridanus resulted in significant inhibition of mycelial growth and yield loss. This study was novel in reporting the vertical transmission of OMSV through basidiospores, and its infection and pathogenicity in a new host P. floridanus.

Nutritional upgrade of olive mill stone waste, walnut shell and their mixtures by applying solid state fermentation initiated by Pleurotus ostreatus.

Present study concerns the transformation of the agro-industrial by-products olive mill stone waste (OMSW) and walnut shell (WS) to a protein-enriched animal feedstuff utilizing the solid state fermentation (SSF) technique. For this purpose, various mixtures of these by-products were exploited as substrates of the SSF process which was initiated by the P. ostreatus fungus. The respective results indicated that the substrate consisted of 80% WS and 20% OMSW afforded the product with the highest increase in protein content, which accounted the 7.57% of its mass (69.35% increase). In addition, a 26.13% reduction of lignin content was observed, while the most profound effect was observed for their 1,3-1,6 ß-glucans profile, which was increased by 3-folds reaching the 6.94% of substrate's mass. These results are indicative of the OMSW and WS mixtures potential to act as efficient substrate for the development of novel proteinaceous animal feed supplements using the SSF procedure. Study herein contributes to the reintegration of the agro-industrial by-products aiming to confront the problem of proteinaceous animal feed scarcity and reduce in parallel the environmental footprint of the agro-industrial processes within the context of circular economy.

In vitro digestibility and angiotensin converting enzyme (ACE) inhibitory activity of solid-state fermented fava beans (Vicia faba L.).

Reducing the content of quickly absorbed carbohydrates and saturated fats in snack formulations while increasing the consumption of high-quality proteins are effective strategies to prevent obesity in childhood. Thus, the nutritional value, digestibility, and functionality of fava beans (Vicia faba L.) fermented with Pleurotus ostreatus were examined as potential ingredients for food design. Solid-state fermentation enhanced the protein content by 16% with a rise in essential (25%) and non-essential (15%) amino acids while decreasing total carbohydrate content and tannin levels. Moreover, fermentation modified the amino acid profile released during digestion, increasing amino acids such as valine, isoleucine, and threonine, which are vital for health and development in childhood. Furthermore, the bioaccessible fraction of the fermented bean showed a 60% of ACE inhibition and improved magnesium bioaccessibility. Consequently, fava beans fermented with Pleurotus ostreatus emerged as a new ingredient in the development of new protein-rich snacks tailored for children and adolescents.

Effects of different selenium biofortification methods on Pleurotus eryngii polysaccharides: Structural characteristics, antioxidant activity and binding capacity in vitro.

The effects of selenium biofortification methods involving sodium selenite and selenium yeast on the structural characteristics, antioxidant activity and binding capacity of Pleurotus eryngii polysaccharides were investigated. Sodium selenite Se-enriched Pleurotus eryngii polysaccharides (Se-SPEP), selenium yeast Se-enriched Pleurotus eryngii polysaccharides (Se-YPEP), and Pleurotus eryngii polysaccharides (PEP) had Se contents of 20.548 ± 1.561, 19.822 ± 0.613, and 0.052 ± 0.016 µg/g, respectively. Compared with PEP, Se-SPEP and Se-YPEP had lower molecular weight and contained the same monosaccharides in varying molar ratios. The results of FT-IR, PS, ZP, and SEM indicated significant alterations in structural characteristics following selenium biofortification. Se-PEPs exhibited superior activity against ABTS, DPPH, and ·OH radicals, as well as the higher binding capacity for Cd2+ and Cu2+ compared to natural polysaccharides. The binding capacity of the polysaccharides for Cd2+ and Cu2+ was higher at pH 6.8 compared to pH 2.0, while the opposite was observed for Pb2+. Furthermore, Se-PEPs exhibited a significantly higher binding capacity for Cd2+ and Cu2+ at both pH levels compared to natural polysaccharides (P < 0.05). Se-YPEP displayed higher antioxidant activity than Se-SPEP, with their binding capacities reversed. These data indicated that selenium biofortification methods have different positive impacts on the structure and activity of polysaccharides compared to natural polysaccharides, making Se-PEPs promising dietary supplements for safeguarding the body against the risks posed by food-derived heavy metals.

PoSnf1 affects cellulose utilization through interaction with cellobiose transporter in Pleurotus ostreatus.

Pleurotus ostreatus is one of the most cultivated edible fungi worldwide, but its lignocellulose utilization efficiency is relatively low (<50 %), which eventually affects the biological efficiency of P. ostreatus. Improving cellulase production and activity will contribute to enhancing the lignocellulose-degrading capacity of P. ostreatus. AMP-activated/Snf1 protein kinase plays important roles in regulating carbon and energy metabolism. The Snf1 homolog (PoSnf1) in P. ostreatus was obtained and analyzed using bioinformatics. The cellulose response of PoSnf1, the effect of the phosphorylation level of PoSnf1 on the expression of cellulose degradation-related genes, the putative proteins that interact with the phosphorylated PoSnf1 (P-PoSnf1), the cellobiose transport function of two sugar transporters (STP1 and STP2), and the interactions between PoSnf1 and STP1/STP2 were studied in this research. We found that cellulose treatment improved the phosphorylation level of PoSnf1, which further affected cellulase activity and the expression of most cellulose degradation-related genes. A total of 1, 024 proteins putatively interacting with P-PoSnf1 were identified, and they were enriched mainly in the substances transport and metabolism. Most of the putative cellulose degradation-related protein-coding genes could respond to cellulose. Among the P-PoSnf1-interacting proteins, the functions of two sugar transporters (STP1 and STP2) were further studied, and the results showed that both could transport cellobiose and were indirectly regulated by P-PoSnf1, and that STP2 could directly interact with PoSnf1. The results of this study indicated that PoSnf1 plays an important role in regulating the expression of cellulose degradation genes possibly by affecting cellobiose transport.

Evaluation of Physicochemical Properties and Prebiotics Function of a Bioactive Pleurotus eryngii Aqueous Extract Powder Obtained by Spray Drying.

A bioactive Pleurotus eryngii aqueous extract powder (SPAE) was obtained by spray drying and its performance in terms of physicochemical properties, in vitro digestion, inflammatory factors, and modulation of the intestinal microbiota was explored. The results indicated that the SPAE exhibited a more uniform particle size distribution than P. eryngii polysaccharide (PEP). Meanwhile, a typical absorption peak observed at 843 cm-1 in the SPAE FTIR spectra indicated the existence of α-glycosidic bonds. SPAE exhibited higher antioxidant abilities and superior resistance to digestion in vitro. In addition, SPAE supplementation to mice significantly reduced the release of factors that promote inflammation, enhanced the secretion of anti-inflammatory factors, and sustained maximum production of short-chain fatty acids (SCFAs). Additionally, it significantly enhanced the relative abundance of SCFAs-producing Akkermansia and reduced the abundance of Ruminococcus and Clostridiides in intestines of mice. These results show the potential of SPAE as a novel material with prebiotic effects for the food and pharmaceutical industries.

Regulation of dye-decolorizing peroxidase gene expression in Pleurotus ostreatus grown on glycerol as the carbon source.

Dye-decolorizing peroxidases (DyPs) (E.C. 1.11.1.19) are heme peroxidases that catalyze oxygen transfer reactions similarly to oxygenases. DyPs utilize hydrogen peroxide (H2O2) both as an electron acceptor co-substrate and as an electron donor when oxidized to their respective radicals. The production of both DyPs and lignin-modifying enzymes (LMEs) is regulated by the carbon source, although less readily metabolizable carbon sources do improve LME production. The present study analyzed the effect of glycerol on Pleurotus ostreatus growth, total DyP activity, and the expression of three Pleos-dyp genes (Pleos-dyp1, Pleos-dyp2 and Pleos-dyp4), via real-time RT-qPCR, monitoring the time course of P. ostreatus cultures supplemented with either glycerol or glucose and Acetyl Yellow G (AYG) dye. The results obtained indicate that glycerol negatively affects P. ostreatus growth, giving a biomass production of 5.31 and 5.62 g/L with respective growth rates (micra; m) of 0.027 and 0.023 h-1 for fermentations in the absence and presence of AYG dye. In contrast, respective biomass production levels of 7.09 and 7.20 g/L and growth rates (µ) of 0.033 and 0.047 h-1 were observed in equivalent control fermentations conducted with glucose in the absence and presence of AYG dye. Higher DyP activity levels, 4,043 and 4,902 IU/L, were obtained for fermentations conducted on glycerol, equivalent to 2.6-fold and 3.16-fold higher than the activity observed when glucose is used as the carbon source. The differential regulation of the DyP-encoding genes in P. ostreatus were explored, evaluating the carbon source, the growth phase, and the influence of the dye. The global analysis of the expression patterns throughout the fermentation showed the up- and down- regulation of the three Pleos-dyp genes evaluated. The highest induction observed for the control media was that found for the Pleos-dyp1 gene, which is equivalent to an 11.1-fold increase in relative expression (log2) during the stationary phase of the culture (360 h), and for the glucose/AYG media was Pleos-dyp-4 with 8.28-fold increase after 168 h. In addition, glycerol preferentially induced the Pleos-dyp1 and Pleos-dyp2 genes, leading to respective 11.61 and 4.28-fold increases after 144 h. After 360 and 504 h of culture, 12.86 and 4.02-fold increases were observed in the induction levels presented by Pleos-dyp1 and Pleos-dyp2, respectively, in the presence of AYG. When transcription levels were referred to those found in the control media, adding AYG led to up-regulation of the three dyp genes throughout the fermentation. Contrary to the fermentation with glycerol, where up- and down-regulation was observed. The present study is the first report describing the effect of a less-metabolizable carbon source, such as glycerol, on the differential expression of DyP-encoding genes and their corresponding activity.

Taxonomic Identification and Nutritional Profiling of Wild Isolates of the Pink Oyster Mushroom Pleurotus djamor (Agaricomycetes) from Northwestern Himalayas (India).

For generations, individuals have been incorporating wild edible fungi into their diets. Precisely identifying mushrooms is essential for harnessing their potential benefits for human use. Specifically, the taxonomical identification of Pleurotus species is known for its complexity. In modern times, it is always necessary to use molecular analysis as a valuable approach for gaining a deeper understanding of the systematics and the delineation of species. Thus, this study confirmed the identity of two wild isolates of Pleurotus djamor (pink oyster mushroom) using the internal transcribed spacer sequences. The amplicons underwent sequencing, and the resulting data was assembled and analyzed. The nucleotide sequences of the two wild isolates blasted against sequences from GenBank database revealed that two wild isolates matched 96.80 and 95.27% P. djamor with accession number KF280324.1 and KT273366.1. Commercially cultivated species took less time for spawn run (17.75 days), primordial initiation (22.25 days) and recoded highest biological efficiency (73.92%) among wild isolates. Nutritional analysis revealed that wild isolates showcase an abundance of nutrients, surpassing commercially cultivated species in terms of moisture, protein, and crude fiber content. This distinctive nutritional profile positions wild isolates as an exceptional superfood, fostering overall health enhancement.

Dietary incorporation of brown seaweed spent oyster mushroom substrate alters growth performance, physiological responses and meat quality parameters in Boschveld roosters.

Use of brown seaweed (Ecklonia maxima) as a nutraceutical source in indigenous chicken diets is limited by high dietary fibre levels. Inoculating seaweeds with oyster mushroom (Pleurotus ostreatus) spawn (OMS) could enhance the utility of the spent mushroom substrate (SMS). This study investigated the effect of feeding incremental levels of brown seaweed SMS on growth performance, physiological responses, and meat quality parameters in Boschveld roosters. A total of 324, 4-week-old Boschveld roosters were weighed and randomly allotted to 36 pens (9 birds per pen) to produce six replicates per dietary treatment. The diets were formulated as follows: a standard grower diet (CON); and CON containing 150 g/kg of brown seaweed inoculated with OMS at 0 (SMS0), 20 (SMS20), 30 (SMS30), 40 (SMS40) and 50% (SMS50). Birds fed diet CON had the least feed intake (p < 0.05) than all the other SMS treatment levels in weeks 7, 8, 12, 14 and 15. Diet SMS40 promoted higher (p < 0.05) body weight gain (BWG) than CON in weeks 6, 7, 9 and 14. Gain-to-feed ratio linearly increased in weeks 7 [R2 = 0.288; p = 0.010], 11 [R2 = 0.581, p = 0.0001] and 14 [R2 = 0.389, p = 0.004], respectively. Quadratic responses (p < 0.05) were observed for BWG in week 5, white blood cells, heterophils, platelets, lymphocytes, monocytes, and relative spleen and large intestine weights as OMS levels increased. Linear increases were recorded for slaughter [R2 = 0.197, p = 0.017] and breast weights [R2 = 0.197, p = 0.020] as OMS levels increased. Diet SMS0 promoted higher (p < 0.05) relative caeca weights than the CON and SMS treatment groups. Neither quadratic nor linear responses (p > 0.05) were observed for breast meat quality parameters. In conclusion, feeding brown seaweed SMS improved growth performance and slaughter weight, altered some blood parameters and internal organs, without affecting breast meat quality of Boschveld roosters. Based on the quadratic response for BWG, the optimum OMS level was deduced at 20% in a brown seaweed-based Boschveld rooster diet.

Pleurotus sajor-caju (Fr.) Singer ß-1,3-Glucanoligosaccharide (Ps-GOS) Suppresses RANKL-Induced Osteoclast Differentiation and Function in Pre-Osteoclastic RAW 264.7 Cells by Inhibiting the RANK/NFκB/cFOS/NFATc1 Signalling Pathway.

Edible grey oyster mushroom, Pleurotus sajor-caju, ß (1,3), (1,6) glucan possesses a wide range of biological activities, including anti-inflammation, anti-microorganism and antioxidant. However, its biological activity is limited by low water solubility resulting from its high molecular weight. Our previous study demonstrated that enzymatic hydrolysis of grey oyster mushroom ß-glucan using Hevea ß-1,3-glucanase isozymes obtains a lower molecular weight and higher water solubility, Pleurotus sajor-caju glucanoligosaccharide (Ps-GOS). Additionally, Ps-GOS potentially reduces osteoporosis by enhancing osteoblast-bone formation, whereas its effect on osteoclast-bone resorption remains unknown. Therefore, our study investigated the modulatory activities and underlying mechanism of Ps-GOS on Receptor activator of nuclear factor kappa-Β ligand (RANKL) -induced osteoclastogenesis in pre-osteoclastic RAW 264.7 cells. Cell cytotoxicity of Ps-GOS on RAW 264.7 cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and its effect on osteoclast differentiation was determined by tartrate-resistant acid phosphatase (TRAP) staining. Additionally, its effect on osteoclast bone-resorptive ability was detected by pit formation assay. The osteoclastogenic-related factors were assessed by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), Western blot and immunofluorescence. The results revealed that Ps-GOS was non-toxic and significantly suppressed the formation of mature osteoclast multinucleated cells and their resorption activity by reducing the number of TRAP-positive cells and pit formation areas in a dose-dependent manner. Additionally, Ps-GOS attenuated the nuclear factor kappa light chain-enhancer of activated B cells' P65 (NFκB-P65) expression and their subsequent master osteoclast modulators, including nuclear factor of activated T cell c1 (NFATc1) and Fos proto-oncogene (cFOS) via the NF-κB pathway. Furthermore, Ps-GOS markedly inhibited RANK expression, which serves as an initial transmitter of many osteoclastogenesis-related cascades and inhibited proteolytic enzymes, including TRAP, matrix metallopeptidase 9 (MMP-9) and cathepsin K (CTK). These findings indicate that Ps-GOS could potentially be beneficial as an effective natural agent for bone metabolic disease.

Pleurotus eryngii polysaccharides alleviate aflatoxin B1-induced liver inflammation in ducks involving in remodeling gut microbiota and regulating SCFAs transport via the gut-liver axis.

Aflatoxin B1 (AFB1) is one of the most widespread contaminants in agricultural commodities. Pleurotus eryngii (PE) is widely used as a feed additive for its anti-inflammatory properties, and its major active substance is believed to be polysaccharides. This study aims to explore the underlying mechanism of dietary PE polysaccharides alleviating AFB1-induced toxicity in ducks. The major monosaccharide components of PE polysaccharides were identified as glucose, mannose, galactose, glucuronic acid, and fucose. The results showed that dietary PE polysaccharides could alleviate liver inflammation, alleviate intestinal barrier dysfunction, and change the imbalanced gut microbiota induced by AFB1 in ducks. However, PE polysaccharides failed to exert protective roles on the liver and intestine injury induced by AFB1 in antibiotic-treated ducks. The PE + AFB1-originated microbiota showed a positive effect on intestinal barrier and inflammation, the SCFAs transport via the gut-liver axis, and liver inflammation compared with the AFB1-originated microbiota in ducks. These findings provided a possible mechanism that PE polysaccharides alleviated AFB1-induced liver inflammation in ducks by remodeling gut microbiota, regulating microbiota-derived SCFAs transport via the gut-liver axis, and inhibiting inflammatory gene expressions in the liver, which may provide new insight for therapeutic methods against AFB1 exposure in animals.

Use of Ozone as a Substrate Treatment for the Control of Trichoderma in the Production of Pleurotus ostreatus.

Pleurotus ostreatus is one of the most widely cultivated species in the world. It can be produced in many lignocellulosic substrates after carrying out a treatment to eliminate competing microorganisms. The most commonly used is pasteurization by steam or by immersion in hot water. The aim of this work is to evaluate if ozone can be employed as treatment for decontamination of the substrate used for the production of the edible mushroom P. ostreatus to control of green mold Trichoderma. Wheat straw was employed as a substrate. We used two different methodologies: bubbling ozone into a tank with water and the substrate, and injecting ozone into a closed tank with the substrate inside. Ten treatments were carried out including two treatments with inoculation by a spray of conidia of Trichoderma. The effect of ozone on the conidia was also evaluated. We found that the treatment of the substrate with ozone in immersed water resulted more effective (lower growth of Trichoderma) than injecting ozone into a closed tank. Anyway, we found that the contaminant fungi could grow on the substrate in both treatments with ozone. We observed that although ozone affected the conidia when it was bubbled into water, some of them still managed to survive and could germinate 72 h later. P. ostreatus could grow and produce fruiting bodies on a substrate that was previously treated with ozone and yields were not affected. Based on the results obtained, we conclude that ozone may not be an effective agent to control Trichoderma in highly contaminated substrates, at least in the experimental conditions that we used, for the production of P. ostreatus.