Subsequent Cultivation Research Articles

The impact of pre-analytical treatment and sorting on human neutrophil function

This study was designed to test the hypothesis that pre-analytical treatment of relatively fragile cells is more important for their viability and function than shear forces and decompression shocks on a flow cytometric sorter. Human neutrophils were enriched using different techniques (sedimentation, hypotonic or ammonium chloride-mediated erythrocyte lysis) and their function was evaluated by measuring their capability of mounting oxidative burst. In contrast to other isolation techniques, neutrophils enriched by spontaneous sedimentation were found to be intact both in terms of their function and relative numbers within the leukocyte population; such preparations were thus run on a FACSAria cell sorter using 70, 85 and 100 µm nozzles and the corresponding internal pressure values. The viability and function of sorted neutrophils with or without subsequent cultivation were re-evaluated using phagocytosis assays. Independent of the nozzle size and internal sorter pressure, the capability of neutrophils of responding to E. coli stimulation was impaired after sorting while phorbol myristate acetate stimulation remained intact. In tested samples, only 20 h incubation of sorted cells confirmed the expected influence of the nozzle size on the sorted cell function. We conclude that pre-analytical treatment is more important for cell function than conditions during sorting.

Influence of ultrahigh frequency irradiation on Photobacterium phosphoreum luxb gene expression

AbstractContinuous increase in the number and the variety of anthropogenic sources of electromagnetic radiation causes a high interest in studying the effects ultrahigh frequency on living organisms. In the present research influence of UHF EMR (15 W, 2.45 GHz) for 5 and 15 min on morphological and genetic peculiarities of Photobacterium phosphoreum colonies was studied. It has been revealed that UHF EMR affected colony growth parameters, induced transcriptional activity of luciferase encoding gene expression and that the effect was depended on exposure duration. The subsequent cultivation of bacteria during a two week period after treatment showed maintaining of the increased luxb mRNA level in irradiated colonies. Opposite bacterial stress responses were detected to UHF EMR and elevated temperature treatments that assumed UHF EMR comprised of not only thermal but specific component of non-thermal nature.

Positive selection of Wharton's jelly-derived CD105+ cells by MACS technique and their subsequent cultivation under suspension culture condition: A simple, versatile culturing method to enhance the multipotentiality of mesenchymal stem cells

ObjectiveWharton's jelly (WJ), an appropriate source of mesenchymal stem cells (MSCs), has been shown to have a wide array of therapeutic applications. However, the WJ-derived MSCs are very heterogeneous and have limited expression of pluripotency markers. Hence, improvement of their culture condition would promote the efficiency of WJ-MSCs. This study aims to employ a simple method of cultivation to obtain WJ-MSCs which express more pluripotency markers.MethodsCD105+ cells were separated by magnetic-associated (activated) cell sorting from umbilical cord mucous tissue. CD105+ cells were added to Methocult medium diluted in α-minimum essential medium (α-MEM) and seeded in poly(2-hydroxyethyl methacrylate) (poly-HEMA)-coated plates for suspension culture preparation. Differentiation capacity of isolated cells was evaluated in the presence of differentiation-inducing media. The expression of pluripotency markers such as Oct3/4, Nanog, and Sox2 was also analyzed by RT-PCR and western blot techniques. Moreover, immunocytochemistry was performed to detect alpha-smooth muscle actin (antigene) (α-SMA) protein.ResultsWJ-MSCs grew homogeneously and formed colonies when cultured under suspension culture conditions (Non-adhesive WJ-MSCs). They maintained their growth ability in both adherent and suspension cultures for several passages. Non-adhesive WJ-MSCs expressed Oct3/4, Nanog, and Sox2 both at transcriptional and translational levels in comparison to those cultured in conventional adherent cultures. They also expressed α-SMA protein.DiscussionIn this study, we isolated WJ-MSCs using a slightly modified culture condition.Our simple non-genetic method resulted in a homogeneous population of WJ-MSCs, which highly expressed pluripotency markers.ConclusionIn the future, more multipotent WJ-MSCs can be harnessed as a non-embryonic source of MSCs in MSC-based cell therapy.

Lipid accumulation by pelletized culture of Mucor circinelloides on corn stover hydrolysate.

Microbial oil accumulated by fungal cells is a potential feedstock for biodiesel production, and lignocellulosic materials can serve as the carbon source to support the fungal growth. The dilute acid pretreatment of corn stover can effectively break down its lignin structure, and this process generates a hydrolysate containing mostly xylose at very dilute concentration and numerous by-products that may significantly inhibit the cell growth. This study utilized corn stover hydrolysate as the culture media for the growth of Mucor circinelloides. The results showed that Mucor cells formed pellets during the cell growth, which facilitates the cell harvest from dilute solution. The results also showed that the inhibitory effect of furfural, 5-hydroxymethylfurfural (HMF), and acetic acid could be avoided if their concentration was low. In fact, all these by-products may be assimilated as carbon sources for the fungal growth. The results proved the feasibility to reuse the cultural broth water for acid pretreatment and then use for subsequent cell cultivation. The results will have a direct impact on the overall water usage of the process.

Evaluation of Crop Production Practices by Farmers in Tshakhuma, Tshiombo and Rabali Areas in Limpopo Province of South Africa

Limpopo Province is characterised by high climatic variability. This is a serious problem in Limpopo Province considering the fact that the province is in a semi-arid area with low, unreliable rainfall. The rainfall distribution pattern, for example, in the Vhembe district is characterised by wet and dry periods depending on the geographical location. In the Vhembe district high rainfall is usually experienced in the Tshakhuma and Levubu areas. Most of the rainfall received in the Vhembe district is in the form of thunderstorms and showers, and this makes rainfall in the district vary considerably. The impact of lower rainfall has negative effects on the agricultural sector, low rainfall resulting in decreases in agricultural activities, loss of livestock, shortage of drinking water, low yields and shortage of seeds for subsequent cultivation. For example, farmers in Rabali area are supposed to use hybrid seeds due to lack of sufficient irrigation water and also poor rainfall distribution compared to farmers in areas such as Tshakhuma and Tshiombo areas.

Immobilization of microalgae on exogenous fungal mycelium: A promising separation method to harvest both marine and freshwater microalgae

Although various methods have been developed for microalgae harvesting, problems such as chemical toxicity, low efficacy, low flexibility, and high cost, still remain with the current methods. In the present study, an effective biological separation method has been developed to harvest microalgae via immobilization on exogenous fungal mycelium. Fungal mycelium was added into microalgae culture and air mixed for the immobilization. The immobilized microalgae with fungal mycelium were naturally precipitated within 10min after stopping the mixing. Both marine (e.g. Nannochloropsis sp.) and freshwater (e.g. Chlorella vulguris) microalgae were almost completely (94–97%) precipitated using the mycelium of Aspergillus nomius CCK-PDA 7#6. The precipitated mixed biomass was separated by simple filtration using sieve. A 70% initial nutrients supplement to de-algated culture medium gave high growth yield (0.43g/l) in subsequent microalgae cultivation. The mycelium can be obtained free or at low-cost as waste from a fungal fermentation process of producing valuable products. The developed method is, therefore, very promising for economical harvesting of microalgae.

Virulence of in vivo and in vitro produced conidia of Metarhizium brunneum strains for control of wireworms

Wireworms are the soil inhabiting larvae of click beetles and can cause severe damage to arable crops such as potatoes (Solanum tuberosum, L.). Several strains of the entomopathogenic fungus Metarhizium brunneum (Petch) are pathogenic to wireworms. In this study, three European strains of M. brunneum were tested in the laboratory against the most damaging wireworm species in Europe, Agriotes lineatus (L.), Agriotes obscurus (L.) and Agriotes sputator (L.). A Swiss strain, isolated from an A. obscurus cadaver, proved to be most effective, killing up to 73% of A. lineatus and 83% A. obscurus individuals, respectively. The median lethal time (LT50) was 21 days post inoculation (dpi) for A. lineatus and 14 dpi for A. obscurus. The strain did not lose virulence through subsequent cultivation on artificial medium and thus seems to be suitable for mass production as a biocontrol agent for wireworm control.

Evaluation of the variability in runoff and sediment loss in successional fallow vegetation of Southern Nigeria

The effects of three different ages of natural fallow vegetation on runoff and sediment loss were investigated in a part of the rainforest zone of Nigeria. Measurements of runoff amount and sediment loss were made for the months of March to November in 2012 rainy season using runoff plots of 40 m<sup>2</sup>. The average runoff amount for the 5-year-old, 3-year-old, and farmland plots were 0.47, 0.26, and 0.41 mm respectively. The average sediment loss on the 5-year-old, 3-year-old, and farmland plots were 209.24, 50.54, and 124.68 kg/ha, respectively. The lowest losses for both runoff and sediment were recorded on the 3-year-old plot, while the 5-year-old plot experienced the highest losses. The variations in runoff and sediment loss among the treatments were significant at P < 0.001. The results evidently showed that rainfall was principally responsible for the erosional losses on all the fallow treatments, and that ground cover (density of herbs) and girth helped to reduce sediment loss on the 3-year-old and farmland surfaces, respectively. The high amount of erosional losses experienced on the 5-year-old fallow than on the 3-year-old fallow and farmland plots imply that fallow that is not adequately protected by ground cover experiences accelerated soil erosion. The continuous loss in topsoil rich in plant nutrients may prolong the optimal capacity of the soil to regain its loss nutrient for subsequent food crop cultivation.

Mixed Trophic State Production Process for Microalgal Biomass with High Lipid Content for Generating Biodiesel and Biogas

Economically feasible and sustainable energy production from microalgae requires optimization of algal growth, maximization of lipid content, and enhancement of biomass conversion into energy. An innovative, mixed trophic state process with high productivity was implemented to generate microalgae with high lipid content for generating biodiesel and biogas. Auxenochlorella protothecoides, a unicellular green alga, was grown phototrophically to 0.28 dry weight per L (gdw/L) then concentrated to 36 gdw/L for use as an inoculum for a subsequent heterotrophic cultivation to a final density of nearly 120 gdw/L. Simultaneous nitrogen deprivation and glucose supplementation during the heterotrophic stage increased the total lipid content from 16 to 57 % while the triacylglycerol (TAG) fraction of total lipids advanced from 2 to 79 %. Productivity peaked at 4.9 g of biomass/L-h and 1.7 g TAGs/L-h. The extracted lipids, including high levels of oleic, linoleic, and palmitic acids, were converted into biodiesel with a predicted cetane number of 56.4 and low concentrations of long-chain saturated and polyunsaturated fatty acid methyl esters. Both intact microalgal biomass and lipid-extracted algal residues (LEA) were good substrates for anaerobic digestion (AD) with methane yields of 0.6 and 0.4 L/g volatile solids (VS), respectively. These yields represented nearly 80 % of theoretical methane potential. LEA, with a favorable carbon to nitrogen ratio (C:N) of approximately 19:1, is an appropriate substrate for anaerobic microorganisms, most likely because it contains essential nutrients required for microbial digestion. The biochemical composition of the biomass, especially its lipid content, is the major contributor for energy output. As a result, coupling biodiesel production with AD of LEA to generate methane can increase the overall process’ energy output up to 40 %.

Phanerochaete chrysosporium pretreatment of biomass to enhance solvent production in subsequent bacterial solid-substrate cultivation

This study investigated pretreatment of corn stover using solid substrate cultivation (SSC) of Phanerochaete chrysosporium (P. chrysosporium) to improve subsequent accessibility to cellulose. Thereafter, Clostridium thermocellum (C. thermocellum) was directly inoculated onto the pretreated biomass to accomplish hydrolysis, followed by solventogenesis by introducing Clostridium beijerinckii (C. beijerinckii).An enzyme suite containing laccase, lignin peroxidase and manganese peroxidase activity was detected during the cultivation of P. chrysosporium on corn stover within 288 h at an initial moisture content wH2O = 80% (mass of water/total mass). Incubation factors, such as substrate moisture content and cultivation temperature affected the percent of lignin removal which ranged from 14.4% to 36.4% of the original lignin. Lignin removal increased as the cultivation of P. chrysosporium continued but was accompanied by increased cellulose loss. The 7-day fungal cultivation sufficiently delignified the corn stover for the subsequent processing. Approximately 25% of the original lignin was removed; however 18% of the initial cellulose was also removed with the lignin. The investigations of the effect of fungal pretreatment were extended to miscanthus, wheat straw and switch grass. The yield of reducing sugar produced by C. thermocellum on pretreated biomass was doubled compared with non-pretreated biomass, demonstrating that pretreatment resulted in a more accessible carbon source for the solvent-producing bacterium. The final comprehensive comparison between the pretreated biomass and non-pretreated biomass on the three-stage SSC for butanol production showed pretreatment by P. chrysosporium improved microbial utilization of lignocellulosic materials for solvent production by approximate 4–7 folds.

One-Step Surgery With Multipotent Stem Cells for the Treatment of Large Full-Thickness Chondral Defects of the Knee

Background: Chondral lesions in athletically active patients cause considerable morbidity, and treatment with existing cell-based therapies can be challenging. Bone marrow has been shown as a possible source of multipotent stem cells (MSCs) with chondrogenic potential and is easy to harvest during the same surgical procedure. Purpose: To investigate the clinical outcome in a group of active patients with large full-thickness chondral defects of the knee treated with 1-step surgery using bone marrow–derived MSCs and a second-generation matrix. Study Design: Case series; Level of evidence, 4. Methods: From January 2007 to February 2010, 25 patients (average age, 46.5 years) with symptomatic large chondral defects of the knee (International Cartilage Repair Society grade 4) who underwent cartilage transplantation with MSCs and a collagen type I/III matrix were followed up for a minimum of 3 years. The average lesion size was 8.3 cm2. Coexisting injuries were treated during the same surgical procedure in 18 patients. All patients underwent a standard postoperative rehabilitation program. Preoperative and postoperative evaluations at 1-year, 2-year, and final follow-up included radiographs, magnetic resonance imaging (MRI), and visual analog scale (VAS) for pain, International Knee Documentation Committee (IKDC), Knee injury and Osteoarthritis Outcome Score (KOOS), Lysholm, Marx, and Tegner scores. Seven patients underwent second-look arthroscopic surgery, with 4 consenting to a tissue biopsy. Results: No patients were lost at final follow-up. The average preoperative values for the evaluated scores were significantly improved at final follow-up (P < .001): VAS, 5.4 ± 0.37 to 0.48 ± 0.19; IKDC subjective, 37.92 ± 4.52 to 81.73 ± 2.42; KOOS pain, 61.04 ± 3.95 to 93.32 ± 1.92; KOOS symptoms, 55.64 ± 3.23 to 89.32 ± 2.32; KOOS activities of daily living, 63.96 ± 4.48 to 91.20 ± 2.74; KOOS sports, 34.20 ± 5.04 to 80.00 ± 3.92; KOOS quality of life, 32.20 ± 4.43 to 83.04 ± 3.37; Lysholm, 46.36 ± 2.25 to 86.52 ± 2.73; Marx, 3.00 ± 0.79 to 9.04 ± 0.79; and Tegner, 2.12 ± 0.32 to 5.64 ± 0.26. Patients younger than 45 years of age and those with smaller or single lesions showed better outcomes. The MRI scans showed good stability of the implant and complete filling of the defect in 80% of patients, and hyaline-like cartilage was found in the histological analysis of the biopsied tissue. No adverse reactions or postoperative complications were noted. Conclusion: The treatment of large chondral defects with MSCs is an effective procedure and can be performed routinely in clinical practice. Moreover, it can be achieved with 1-step surgery, avoiding a previous surgical procedure to harvest cartilage and subsequent chondrocyte cultivation.

Forest Continuity as a Key Determinant of Soil Carbon and Nutrient Storage in Beech Forests on Sandy Soils in Northern Germany

Forest (or tree) age has been identified as an important determinant of the carbon (C) storage potential of forest soils. A large part of Central Europe’s current forested area was affected by land use change with long periods of cultivation in past centuries suggesting that the organic C stocks in the soil (SOC) under recent forest may partly be legacies of the past and that stand age effects have to be distinguished from forest continuity effects (that is, the time since re-afforestation). We examined the influence of mean tree age and forest continuity on the SOC pool and the stores of total N and available P, Ca, Mg, and K in the soil (mineral soil and organic layer) across a sample of 14 beech (Fagus sylvatica) forests on sandy soil with variable tree age (23–189 years) and forest continuity (50-year-old afforestation to ancient (‘permanent’) forest, that is, >230 years of proven continuity). Ancient beech forests (>230 years of continuity) stored on average 47 and 44% more organic C and total N in the soil than recent beech afforestation (50–128 years of continuity). Contrary to expectation, we found large and significant C and N pool differences between the forest categories in the mineral soil but not in the organic layer indicating that decade- or century-long cultivation has reduced the subsoil C and nutrient stores while the organic layer element pools have approached a new equilibrium after only 50–128 years. PCA and correlation analyses suggest that forest continuity cannot be ignored when trying to understand the variation in soil C stocks between different stands. Forest clearing, subsequent cultivation, and eventual re-afforestation with beech resulted in similar relative stock reductions of C and N and, thus, no change in soil C/N ratio. We conclude that the continuity of forest cover, which may or may not be related to tree age, is a key determinant of the soil C and nutrient stores of beech forests in the old cultural landscape of Central Europe.

Phylogenetic characterization and quantification of ammonia-oxidizing archaea and bacteria from Lake Kivu in a long-term microcosm incubation.

A microcosm cultivation-based method was set up to investigate the growth of ammonia-oxidizing archaea (AOA), isolated from a water sample acquired at a depth of 50 m from the northern basin of Lake Kivu. For this purpose, both CARD-FISH and qPCR targeting of archaeal 16S rRNA and amoA genes were used. Archaeal cell growth at the end of the 246-day microcosm experiment accounted for 35% of the SybrGold-stained cells, which corresponded to 6.61 x 10(6) cells/ml and 1.76 +/- 0.09 x 10(6) archaeal 16S rRNA gene copies/ml. Clone libraries and DGGE fingerprinting confirmed the dominance of AOA phylotypes in the archaeal community microcosm. The majority of the identified archaeal 16S rRNA gene sequences in the clone libraries were affiliated with Thaumarchaeota Marine Group 1 .1a. Subsequent cultivation of the AOA community on deep-well microtiter plates in medium containing different carbon sources to stimulate archaeal growth failed to show significant differences in archaeal abundance (ANOVA t14 = -1.058, P = 0.308 and ANOVA t14= 1.584, P = 0.135 for yeast extract and simple organic acids, respectively). The lack of growth stimulation by organic compounds is in concordance with the oligotrophic status of Lake Kivu. Finally, the addition of antibiotics to the growth medium resulted in archaeal cell counts that were significantly lower than those obtained from cultures in antibiotic-free medium (ANOVA t14 = 12.12, P < 0.001).

Generation of functional endothelial-like cells from adult mouse germline-derived pluripotent stem cells

Functional endothelial cells and their progenitors are required for vascular development, adequate vascular function, vascular repair and for cell-based therapies of ischemic diseases. Currently, cell therapy is limited by the low abundance of patient-derived cells and by the functional impairment of autologous endothelial progenitor cells (EPCs). In the present study, murine germline-derived pluripotent stem (gPS) cells were evaluated as a potential source for functional endothelial-like cells.Cells displaying an endothelial cell-like morphology were obtained from gPS cell-derived embryoid bodies using a combination of fluorescence-activated cell sorting (FACS)-based selection of CD31-positive cells and their subsequent cultivation on OP9 stromal cells in the presence of VEGF-A. Real-time reverse transcriptase polymerase chain reaction, FACS analysis and immunofluorescence staining showed that the gPS cell-derived endothelial-like cells (gPS-ECs) expressed endothelial cell-specific markers including von Willebrand Factor, Tie2, VEGFR2/Flk1, intercellular adhesion molecule 2 and vascular endothelial-cadherin. The high expression of ephrin B2, as compared to Eph B4 and VEGFR3, suggests an arterial rather than a venous or lymphatic differentiation. Their capability to take up Dil-conjugated acetylated low-density lipoprotein and to form capillary-like networks on matrigel confirmed their functionality.We conclude that gPS cells could be a novel source of endothelial cells potentially suitable for regenerative cell-based therapies for ischemic diseases.

Isolation and identification of the microalgal symbiont from primmorphs of the endemic freshwater sponge Lubomirskia baicalensis (Lubomirskiidae, Porifera)

The first reports on the presence of zoochlorellae within the organelles of mesenchymal cells from freshwater sponges were published in the 19th century. Today, it is well-known that freshwater sponges can be found in association with different endosymbiotic algae. However, until now there has been no detailed information about the endosymbiotic chlorophyll-containing algae in the remarkable endemic green sponges from Lake Baikal. In our study we were able for the first time to isolate and identify endosymbionts from primmorphs cultivated in vitro, and to compare them with those from naturally occurring Lubomirskia baicalensis sponges. Structural as well as molecular biological investigations show that the endosymbiotic alga is a Mychonastes species closely related to M. huancayensis. Another novel aspect of our work was to show that it is possible to use primmorphs of endemic sponges for isolation and subsequent cultivation of their endosymbiotic algae. We employed a simple cold-water (3–4°C) approach for cultivating Mychonastes sp., both within sponge primmorphs and in culture.

The effect of media composition on multiplication of grape rootstocks in vitro

The current demand for in vitro cultures of grape rootstocks, not only for mass production of plants, but also for genetic engineering is evident. The study on micropropagation of grape rootstock genotypes namely Kober 5BB, Kober 125AA and Teleki 5C was performed. The aim of the study was to develop an optimized protocol to obtain large quantity of plant material. Protocol is based on regeneration via organogenesis, considering that grape embryogenic calluses are laborious to establish and the genotype of the regenerated plants can be altered. Using of Driver and Kuniyuki Walnut media for the establishing of proliferating cultures gave better results than Murashige Skoog media in case of all used rootstocks. Subsequent cultivation on modified Murashige Skoog media with 1-naphtalene acetic acid and increased concentration of cytokynin was characterized by multiplication of cultures and formation of clusters with high multiplication capability. The clusters obtained from rootstock genotypes were suitable for mass propagation as well as for genetic transformation due to their high ability of regeneration.

Efeito do neem (Azadirachta Indica) no controle de nematódeos gastrintestinais em ovinos suplementados a pasto no período seco

The aim of this study was to evaluated the effect of Azadirachta indica (Neem) extracts upon the productive response and control of gastrointestinal nematodes in Santa Ines sheep during the dry season grazing. Twenty four sheep, were divided in three groups according to the following treatments: SP – Control (Animals receiving protein supplement); SP- Oil (Animals receiving protein supplement and Neem oil extract) and SP – Leaf (Animals receiving protein supplement and Neem dry leaves). The animals received daily protein supplement, while the oil and the dry leaves treatments were offered at interval of 15 days, a total of 4 doses ministred. Each day of supply of oil and leaves, feces samples were collected (day 0, 15, 30, 45, 60) for egg count per gram and subsequent larval cultivation for identifying genres. The protein supplement directly influenced the reduction of nematode eggs obtained per gram of feces (FEC) of sheep on quality grazing, consequently bringing benefits for the worms control. A indica present in the oil and leaves did not obtain any statistical difference between the treatments when compared to the animals receiving only protein supplement. The association of Neem derivatives with protein supplement have not been able to increase animal performance even after decreasing the rates of FEC, showing that protein supplement were more able to provide effects on internal parasites control, when compared with to Neem oil or Neem dry leaves treatments.

The Value of Satellite-Based Active Fire Data for Monitoring, Reporting and Verification of REDD+ in the Lao PDR

Shifting cultivation is a dominant land-use system in Laos, and fire is the tool commonly used to clear fallow vegetation for subsequent cultivation. We assessed the feasibility of active fire data derived from the Moderate Resolution Imaging Spectroradiometer (MODIS) for monitoring fires in Laos. Specifically, we investigated the potential of the active fire data as input into monitoring, reporting and verification (MRV) systems to assess the effectiveness of measures related to Reducing Emissions from Deforestation and Forest Degradation plus the enhancement of forest carbon stocks (REDD+). Our qualitative and quantitative accuracy assessments of the fire data yielded mixed results with varying degrees of undetected fires and false detections. Hence, at IPCC Tier 3, the uncertainties inherent in the detection accuracy become too large. Active fire data can be valuable for supporting national-level MRV at Tier 2 in combination with auxiliary data for characterizing fire-dependent local land-use systems, such as shifting cultivation.

Enhanced Intensity Dependence as a Marker of Low Serotonergic Neurotransmission in High Optimistic College Students

Positive psychology focuses were on the merits of individuals, such as optimism and positive attitude, and the subsequent cultivation of these virtues. Optimism or pessimism is a significant predictor of physical health outcomes. The present study examined whether optimism or pessimism is associated with the loudness dependence of auditory evoked potentials (LDAEP), a biological indicator of serotonergic neurotransmission, for the N1, P2, and N1/P2 peaks in college students. The amplitudes and amplitude-stimulus intensity function (ASF) slopes of the N1, P2, and N1/P2 peaks were determined in the 24 (10 males) high optimistic and 24 (14 males) high pessimistic individuals. Significantly higher P2 ASF slopes were found in the optimistic group relative to the pessimistic group. Concerning peaks and ASF slopes of N1 and N1/P2, no significant differences were observed. Our results suggest that the serotonergic neurotransmission of the high optimistic college students was inferior to that of the pessimistic ones. Further investigations are needed to provide sufficient support for our results.

Composted versus Raw Olive Mill Waste as Substrates for the Production of Medicinal Mushrooms: An Assessment of Selected Cultivation and Quality Parameters

Two-phase olive mill waste (TPOMW, “alperujo”) is a highly biotoxic sludge-like effluent of the olive-oil milling process with a huge seasonal production. One of the treatment approaches that has so far received little attention is the use of TPOMW as substrate for the cultivation of edible mushrooms. Fifteen fungal strains belonging to five species (Basidiomycota), that is, Agrocybe cylindracea, Pleurotus cystidiosus, P. eryngii, P. ostreatus, and P. pulmonarius, were evaluated for their efficacy to colonize media composed of TPOMW, which was used either raw or composted in mixtures with wheat straw in various ratios. Qualified strains exhibited high values of biological efficiency (e.g., 120–135% for Pleurotus spp. and 125% for A. cylindracea) and productivity in subsequent cultivation experiments on substrates supplemented with 20–40% composted TPOMW or 20% raw TPOMW. Only when supplementation exceeded 60% for raw TPOMW, a negative impact was noted on mushroom yields which could be attributed to the effluent's toxicity (otherwise alleviated in the respective composted TPOMW medium). Earliness and mushroom size as well as quality parameters such as total phenolic content and antioxidant activity did not demonstrate significant differences versus the control wheat-straw substrate. The substrates hemicellulose content was negatively correlated with mycelium growth rates and yields and positively with earliness; in addition, cellulose: lignin ratio presented a positive correlation with mycelium growth and mushroom weight for A. cylindracea and with earliness for all species examined. TPOMW-based media revealed a great potential for the substitution of traditional cultivation substrates by valorizing environmentally hazardous agricultural waste.