Growth Medium Research Articles

Potential Utilization of Defective Fruit Juice Products for Black Soldier Fly Cultivation to Reduce Waste Generation in the Beverage Industry

The waste of fruit juice and tea defective products has organic content that conventionally has the potential to be utilized as a medium or place for maggot growth, but until now it has not been equipped with supporting scientific studies. Therefore, the objectives of the study are: 1) Assessing the characteristics of fruit juice and tea product waste, 2) Assessing the effectiveness of maggot in reducing fruit juice and tea product waste. This research is experimental in nature using variations in the composition of fruit juice and tea product waste as a source of nutrition for maggot which is carried out for 12 days. This variation uses the Complete Randomized Design method with 3 (three) treatments, namely variations in the composition of 60% tea + 40% fruit juice waste (S1), 50% tea + 50% fruit juice waste (S2), and 40% tea + 60% fruit juice waste (S3) in duplo. In addition, this study was completed with a control treatment. The results showed that the characteristics of fruit juice and tea waste containing protein (3.78-4.59%), fat (0.90-1.31%), carbohydrates (28.15-29.15%), crude fiber (2.42-3.12%) can be used as maggot cultivation media. The highest protein and crude fiber contents were found in the S3 composition waste at 4.59% and 3.12%. Maggot cultivation results that showed the highest fat and carbohydrate content were found in S1 composition waste at 1.31% and 29.15%. The recommended composition variation is S1 composition with the highest substrate consumption value and waste reduction index of 57.47% and 20.06%.

Natural Therapeutic Agents’ Efficacy in Preventive Strategies against the Periodontal Pathogen Aggregatibacter actinomycetemcomitans: An In Vitro Study

Adolescent carriers of the Aggregatibacter actinomycetemcomitans JP2 genotype have an increased risk of developing periodontitis, due to the bacterium’s high leukotoxin (LtxA) production. LtxA contributes to marginal bone loss by killing immunity cells, thus activating the proinflammatory interleukin-1β (IL-1β), which, in turn, activates the osteoclasts. A possible strategy to prevent periodontitis might be to neutralize LtxA in JP2-infected individuals. The aim of this study was to investigate whether extracts from Matcha or Guava leaves can prolong the viability of macrophages in cell cultures by neutralizing the highly leukotoxic JP2 genotype bacteria. The A. actinomycetemcomitans JP2 genotype was pretreated with extracts from either Matcha or Guava leaves. Later, the extracts were rinsed off, before JP2 bacteria were exposed to macrophage cell cultures. The experiment was repeated, where JP2 bacteria were persistently treated with the extracts instead, i.e., the extracts were not rinsed off. The macrophage viability after bacterial exposure was analyzed and compared with that of macrophages exposed to untreated JP2 bacteria. IL-1β secretion in the cell culture medium was quantified in all group samples. Pretreatment of the A. actinomycetemcomitans JP2 genotype with Matcha or Guava leaf extracts moderately neutralized LtxA activity, which resulted in prolonged macrophage viability and decreased IL-1β secretion. These effects of prolonged macrophage viability were enhanced when extracts were persistently present during the exposure period. The results indicate that Matcha and Guava leaf extracts have effects on the virulence of the A. actinomycetemcomitans JP2 genotype that may be useful in future treatment strategies to prevent periodontitis in JP2 bacterium carriers.

Response of Staphylococcus aureus physiology and Agr quorum sensing to low-shear modeled microgravity.

Staphylococcus aureus is commonly isolated from astronauts returning from spaceflight. Previous analysis of omics data from S. aureus low Earth orbit cultures indicated significantly increased expression of the Agr quorum sensing system and its downstream targets in spaceflight samples compared to ground controls. In this current study, the rotary cell culture system (RCCS) was used to investigate the effect of low-shear modeled microgravity (LSMMG) on S. aureus physiology and Agr activity. When cultured in the same growth medium and temperature as the previous spaceflight experiment, S. aureus LSMMG cultures exhibited decreased agr expression and altered growth compared to normal gravity control cultures, which are typically oriented with oxygenation membrane on the bottom of the high aspect rotating vessel (HARV). When S. aureus was grown in an inverted gravity control orientation (oxygenation membrane on top of the HARV), reduced Agr activity was observed relative to both traditional control and LSMMG cultures, signifying that oxygen availability may affect the observed differences in Agr activity. Metabolite assays revealed increased lactate and decreased acetate excretion in both LSMMG and inverted control cultures. Secretomics analysis of LSMMG, control, and inverted control HARV culture supernatants corroborated these results, with inverted and LSMMG cultures exhibiting a decreased abundance of Agr-regulated virulence factors and an increased abundance of proteins expressed in low-oxygen conditions. Collectively, these studies suggest that the orientation of the HARV oxygenation membrane can affect S. aureus physiology and Agr quorum sensing in the RCCS, a variable that should be considered when interpreting data using this ground-based microgravity model.IMPORTANCES. aureus is commonly isolated from astronauts returning from spaceflight and from surfaces within human-inhabited closed environments such as spacecraft. Astronaut health and immune function are significantly altered in spaceflight. Therefore, elucidating the effects of microgravity on S. aureus physiology is critical for assessing its pathogenic potential during long-term human space habitation. These results also highlight the necessity of eliminating potential confounding factors when comparing simulated microgravity model data with actual spaceflight experiments.

Growth potential, biochemical properties and nutrient removal efficiency of some freshwater microalgae and their consortia from wastewater

Impact of varying nitrate (NO3-N) and phosphate (PO4-P) concentrations and sewage water (SW) on the growth, nutrient removal, lipid accumulation, enzymatic antioxidant activity and phytochemical contents of the microalgae Scenedesmus dimorphus, Coelastrella tenuitheca, Chroococcus turgidus and Parachlorella kessleri under monoculture and their consortia have been investigated. High growth rates were observed for all the four algae in both mono and mixed culture conditions at enhanced concentrations of N (1500 mg/L NO3-N) and P (40 mg/L PO4-P). The species Scenedesmus dimorphus outperformed other microalgae growing in SW in efficiently removing nitrogen. The algal consortia of mixed species was found to be more effective in phosphorus removal. The carbohydrate and protein contents were highest in Parachlorella kessleri, about 37% and 44%, respectively, in SW cultivation. The algal consortia demonstrated highest starch content (4%) in nitrogen deprived growth medium. Highest lipid production (43%) was observed in the SW culture. The species Coelastrella tenuitheca, Chroococcus turgidus and Scenedesmus dimorphus irrespective of the growth media indicated significant accumulation of phenol, flavonoid and tannin. The DPPH, catalase and ascorbic peroxidase assay showed pronounced antioxidant activity. Nutrient (N and P) enrichment exhibited enhanced antioxidant enzymatic activity and accumulation of cell storage products.

Lactobacilli and Bifidobacteria: A Parapostbiotic Approach to Study and Explain Their Mutual Bioactive Influence

Three strains of Lactiplantibacillus plantarum and three bifidobacteria (Bifidobacterium animalis subsp. lactis, Bifidobacterium breve, and Bifidobacterium subtile) were used as target strains; in addition, for each microorganism, the cell-free supernatant (CFS) was produced and used as an ingredient of the growth medium. Namely CFSs from lactobacilli were used on bifidobacteria and CFSs from bifidobacteria were used on lactobacilli. The viable count was assessed, and the data were modelled through a reparametrized Gompertz equation cast both in the positive and negative form to evaluate the parameters t-7log, which is the time after which the viable count was 7 log CFU/mL, and the t-7log*, which is the time after which the viable count was below 7 log CFU/mL; the difference between the t-7log* and t-7log defines the stability time. Statistics through a multiparametric ANOVA (analysis of variance) provided evidence for the presence of a bifidogenic and/or bioactive factor produced by bifidobacteria and active on lactobacilli, and vice versa (bioactive factor of lactobacilli with a functional effect on bifidobacteria), although further studies are required to better explain the mechanisms beyond the positive effects. In addition, the influence on the target strains can be found during the growth phase (stimulation), as well as during senescence and death phase (protective effect), with a strong strain/species dependence on both CFS production and target strain.

A Microencapsulated Blend Of Botanicals Supports Weaning Piglets During An Lps Challenge By Modulating Liver Inflammation And Intestinal Integrity.

This study examined the action of a blend of botanicals (BOT) against LPS-induced inflammation on cultured hepatocytes and weaning piglets. In vitro studies examined HepG2 cells treated with BOT and challenged with E. coli lipopolysaccharide (LPS) for 8 days. BOT treatment reduced IL-6 concentration in cell culture media across time (p < 0.05) and decreased pro-inflammatory cytokine expression on day 1 and 8 of experiment (TNFα, IL-1β; p < 0.05). BOT also increased the expression of antioxidant enzymes (GPX-2, SOD, CAT) on day 8 (p < 0.05), which was supported by lowered reactive oxygen species concentration after LPS challenge (p < 0.1). The in vivo study was conducted with 72 weaning pigs, allotted into 24 pens and divided into 3 groups: a negative control (CTR-, basal diet), a challenged control (CTR+) that received an intraperitoneal injection of E. coli O55:B5 LPS on day 14 and 16, and a challenged treated group which received a diet containing 1.5g/Kg of microencapsulated BOT (BOT+) for the whole duration of the study. Growth performance was determined weekly and, on days 21 (one animal per pen) and 28 (remaining animals), pigs were sacrificed to collect liver and jejunal tissues. After challenge, BOT+ pigs had increased BW on day 21 (p < 0.05) and 28 (p < 0.1) compared to CTR+. Similar improvements in ADG and FCR on day 14-21 (p < 0.05) and 21-28 (p < 0.1) were also seen in BOT+ group. In the liver, compared to CTR+ pigs, BOT+ pigs downregulated the expression of TLR-4, IL-6, IFN-γ on day 21 (p < 0.05), and TLR-4, TNF-α, IL-8 on day 28 (p < 0.05). BOT+ also increased GPX-2 expression on day 21 and 28 (p < 0.05), while also upregulating SOD-1 and SOD-2 on day 21 (p < 0.05) and CAT on day 28 (p < 0.05) compared to CTR+. In the jejunum, BOT+ reduced inflammation by affecting cytokine expression (p < 0.05) and increasing the expression of tight-junction proteins, ZO-1 on day 21 and CLD-1 on day 28 (p < 0.05). Furthermore, BOT+ pigs had lower crypt depth on day 21 (p < 0.1) and day 28 (p < 0.05), and increased villi to crypt ratio on day 21 and 28 (p < 0.05). By day 28, BOT+ intestinal measurements were restored to values similar to the CTR-. Finally, BOT+ also reduced mast cell activation on day 21 (p < 0.05) compared to CTR+. Considering all the findings, BOT controlled inflammatory activation and the oxidative stress in liver cells, enhanced intestinal integrity, and as a result improved the growth performance of weaning piglets challenged with LPS.

Plant biotechnology for the conservation of Argania spinosa (Skeels L): optimising propagation and sustainable valorisation

ABSTRACT Argania spinosa, an endangered species from southwestern Morocco, is studied for preservation through plant biotechnology. The research aims to optimise argan tree propagation to boost production, preserve heritage, and use residual species. In vitro seedlings are transferred to a modified Murashige and Skoog medium with various phytohormones. Six combinations of auxins and cytokinins are tested for germination. Results show that a liquid medium with activated charcoal from argan shells promotes stem elongation. Meta-topoline at 0.5 mg L−1, combined with naphthalene acetic acid (NAA) at 0.2 mg L−1, enhances shoot growth, leaf formation, and callogenesis. Adding benzylaminopurine (BAP) at 0.1 mg L−1 and indole-3-butyric acid (IBA) at 0.02 mg L−1 to the medium elongates stems by 4.3 cm after 21 days. A 2000 ppm IBA concentration is key for a 90% rooting rate. These findings underscore plant biotechnology’s role in preserving and propagating argan trees, improving resource management, and supporting sustainability.

Anti-inflammation Mechanisms of Flavones Are Highly Sensitive to the Position Isomers of Flavonoids: Acacetin vs Biochanin A.

Acacetin (ACA) and biochanin A (BCA) are isomeric monomethoxyflavones with different structural positions of the 4'-methoxy-phenyl group. Both of them are present in many commonly consumed foods, such as citrus fruits and vegetables, and have been discovered with anti-inflammatory activities, but their mechanisms of action are not clearly elucidated at the molecular level. Herein, we reported the structure-activity relationship of ACA and BCA regarding their potency in inhibiting nitric oxide (NO) production, proinflammatory enzyme expression, and mRNA expression of proinflammatory cytokines in the lipopolysaccharide (LPS)-induced RAW 264.7 cells. Furthermore, transcriptome analysis was conducted to map out the overall pathways impacted by these two isomers. Our results showed that ACA possessed higher suppressive activity in nitric oxide (NO) production (IC50 = 23.93 ± 1.74 μM for ACA and IC50 = 71.41 ± 8.07 μM for BCA), inducible nitric oxide synthase (iNOS) expression, and proinflammatory interleukin (IL)-1β mRNA expression, but lower inhibitory activity in IL-6 mRNA expression as compared with BCA. Although two compounds were revealed to bind to c-Src protein according to drug affinity responsive target stability (DARTS) and western blotting results, molecular docking and molecular dynamics (MD) simulation showed that they had different binding sites, which subsequently resulted in different signaling transduction. ACA primarily exerted anti-inflammatory activity through the mitogen-activated protein kinase (MAPK) signaling pathway, the tumor necrosis factor (TNF) signaling pathway, and the hypoxia-inducible factor (HIF)-1 signaling pathway, while BCA was particularly involved in the Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway, the nuclear factor kappa B (NF-κB) signaling pathway, the TNF signaling pathway, and the toll-like receptor (TLR) signaling pathway. Moreover, two isomers remained stable in the cell culture medium and did not encounter the biotransformation process in RAW 264.7 cells. However, BCA exhibited insufficient cellular uptake ability and transport efficiency in macrophages compared to ACA. Taken together, ACA is more promising for controlling inflammation and worthy of further study for human use.

Poly(3-hydroxybutyrate) production for food packaging from biomass derived carbohydrates by cupriavidus necator DSM 545

The extensive utilization of conventional plastics has resulted in a concerning surge in waste. A potential solution lies in biodegradable polymers mostly derived from renewable sources. Cupriavidus necator DSM 545 is a microorganism capable, under stress conditions, of intracellularly accumulating Poly(3-hydroxybutyrate) (PHB), a bio-polyester. This study aimed to identify optimal conditions to maximize the intracellular accumulation of PHB and its global production using natural media obtained by processing lignocellulosic residues of cardoon, a low-cost feedstock. An intracellular PHB accumulation was observed in all of the tested media, indicating a metabolic stress induced by the lack of macronutrients. Increasing C/N ratios led to a significant decrease in cellular biomass and PHB production. Furthermore C. necator DSM 545 was incapable of consuming more than 25 g/L of supplied monosaccharides. Surprisingly, in the samples supplied with 60 % of the pentose-rich liquid fraction, complete consumption of xylose was observed. This result was also confirmed by subsequent tests using Medium 1 growth media containing xylose as the sole carbon source. Using a diluted medium with a C/N ratio of 5, a PHB production of 5.84 g/L and intracellular PHB accumulation of 77 % w/w were respectively achieved. Finally, comparative shelf-life tests conducted against conventional pre-packaging materials in PP suggested that PHB films performed similarly in preserve ready-to-eat products.

Evaluation of chicken embryo extract and egg yolk extract as alternatives to basic cell culture medium supplement

BackgroundFetal calf serum (FCS), an existing cell culture supplement, is effective but has several drawbacks, including being expensive, requiring a lengthy process of production, and requiring a hard currency. With this in mind, we planned to evaluate chick embryo extract and egg yolk extracts in cell culture as alternatives to fetal calf serum (FCS).MethodsSpecific pathogen-free eggs were purchased from the National Veterinary Institute, Bishoftu, Ethiopia, and incubated in a humidified incubator at 37 °C for 11 days. Egg yolk extract (EYE) and chick embryo extract (CEE) were collected after the egg was opened with caution not to destroy the yolk sack or the chick embryo itself. Chick fibroblasts and Vero cells were cultured in minimum essential medium (MEM) supplemented with egg yolk extract or chick embryo extract at ratios of 0:10, 1:9, 2.5:7.5, and 5:5% fetal calf serum.ResultsFibroblast cell attachment was better in media supplemented with 5% CEE and 5% FCS. The confluency was also greater than 50% at this concentration. Vero cells cultured with 5% CEE and 5% FCS also exhibited very good cell attachment and a confluency of up to 70%. Viability and confluency were also observed at 5%:5% ratios of 50 and 70%, respectively.ConclusionThis investigation evaluated these two extracts as cell culture supplements and revealed promising results as alternatives to fetal calf serum. The limitation of this study is that it only used two cell types and additional cell lines, and different ratios should be tested. With the above findings, further research using different cell lines, ratios and conditions is warranted.

A Novel Recombinant Vitronectin Variant Supports the Expansion and Differentiation of Pluripotent Stem Cells in Defined Animal-Free Workflows

An essential aspect of harnessing the potential of pluripotent stem cells (PSCs) and their derivatives for regenerative medicine is the development of animal-free and chemically defined conditions for ex vivo cultivation. PSCs, including embryonic and induced PSCs (iPSCs), are in the early stages of clinical trials for various indications, including degenerative diseases and traumatic injury. A key step in the workflows generating these cells for more widespread clinical use is their safe and robust ex vivo cultivation. This entails optimization of cell culture media and substrates that are safe and consistent while maintaining robust functionality. Here, we describe the design of a human vitronectin (hVTN) variant with improved manufacturability in a bacterial expression system along with improved function in comparison to wild-type VTN and other previously characterized polypeptide fragments. In conjunction with an animal component-free media formulation, our hVTN fragment provides animal-free conditions for the enhanced expansion of iPSCs. This hVTN variant also supports the reprogramming of PBMCs into iPSCs. Furthermore, we show that these iPSCs can be efficiently differentiated into the three major germ layers and cortical neurons, thereby closing the loop on a completely defined animal-free workflow for cell types relevant for regenerative medicine.

Metabolism of primary high-grade serous ovarian carcinoma (HGSOC) cells under limited glutamine or glucose availability

BackgroundHigh-grade serous ovarian carcinoma (HGSOC) is the most common and aggressive subtype of epithelial ovarian carcinoma. It is primarily diagnosed at stage III or IV when the 5-year survival rate ranges between 20% and 40%. Here, we aimed to validate the hypothesis, based on HGSOC cell lines, that proposed the existence of two distinct groups of HGSOC cells with high and low oxidative phosphorylation (OXPHOS) metabolism, respectively, which are associated with their responses to glucose and glutamine withdrawal.MethodsWe isolated and cultivated primary cancer cell cultures from HGSOC and nontransformed ovarian fibroblasts from the surrounding ovarium of 45 HGSOC patients. We tested the metabolic flexibility of the primary cells, particularly in response to glucose and glutamine depletion, analyzed and modulated endoplasmic reticulum stress, and searched for indices of the existence of previously reported groups of HGSOC cells with high and low OXPHOS metabolism.ResultsThe primary HGSOC cells did not form two groups with high and low OXPHOS that responded differently to glucose and glutamine availabilities in the cell culture medium. Instead, they exhibited a continuum of OXPHOS phenotypes. In most tumor cell isolates, the responses to glucose or glutamine withdrawal were mild and surprisingly correlated with those of nontransformed ovarian fibroblasts from the same patients. The growth of tumor-derived cells in the absence of glucose was positively correlated with the lipid trafficking regulator FABP4 and was negatively correlated with the expression levels of HK2 and HK1. The correlations between the expression of electron transport chain (ETC) proteins and the oxygen consumption rates or extracellular acidification rates were weak. ER stress markers were strongly expressed in all the analyzed tumors. ER stress was further potentiated by tunicamycin but not by the recently proposed ER stress inducers based on copper(II)-phenanthroline complexes. ER stress modulation increased autophagy in tumor cell isolates but not in nontransformed ovarian fibroblasts.ConclusionsAnalysis of the metabolism of primary HGSOC cells rejects the previously proposed hypothesis that there are distinct groups of HGSOC cells with high and low OXPHOS metabolism that respond differently to glutamine or glucose withdrawal and are characterized by ETC protein levels.

GDF-15 predicts epithelioid hemangioendothelioma aggressiveness and is down-regulated by sirolimus through ATF4/ATF5 suppression.

Epithelioid hemangioendothelioma (EHE) poses a therapeutic challenge due to limited efficacy of conventional chemotherapy in advanced cases, necessitating exploration of new treatment avenues and identification of novel aggressiveness biomarkers. This study aimed at i) utilizing an EHE patient-derived xenograft (PDX) model and its associated cell line to assess the efficacy of sirolimus and ii) analyzing two distinct patient cohorts to pinpoint circulating biomarkers of EHE aggressiveness. A PDX model and corresponding cell line were established from an advanced EHE patient, demonstrating consistency with the original tumor in terms of histomorphology, WWTR1::CAMTA1 fusion presence, and genomic and transcriptomic profiles. Two independent patient series were employed to investigate the association between Growth/Differentiation Factor 15 (GDF-15) serum levels and EHE aggressiveness. ELISA analyses on EHE cell culture medium and blood from EHE-carrying mice revealed the release of GDF-15 by EHE cells. Sirolimus exhibited markedly higher anti-tumor activity compared to doxorubicin, concurrently reducing GDF-15 expression/release both in vivo and in vitro. This reduction was attributed to the drug-induced inhibition of phosphorylation/activation of 4E-BP1 and subsequent downregulation of the GDF-15 transcription factors ATF4 and ATF5. Blood sample analyses from two independent patient series showed a significant correlation between GDF-15 and EHE aggressiveness. This study identifies GDF-15 as a novel biomarker of EHE aggressiveness and underscores the superior efficacy of sirolimus compared to doxorubicin in our experimental models. The observed inhibition of GDF-15 release by sirolimus suggests its potential as a biomarker for monitoring the drug's activity in patients.

Characterization of Bacillus pacificus KUMBNGBT-39-Derived Polyhydroxybutyrate and Impact of Physicochemical Factors on its Production.

The extensive use of various chemicals in synthetic plastics is toxic and threatens the biosphere. To address this, the study aimed to isolate, screen, characterize, optimize, and quantify polyhydroxybutyrate (PHB)-producing bacteria using cost-effective residues. Isolated from a landfill site, the Gram-positive, rod-shaped, spore-forming, motile bacterium with intracellular PHB granules was identified as Bacillus pacificus based on phenotypic and genotypic characteristics. Optimal PHB production parameters included a nutrient broth medium, 72h of incubation, a temperature of 37° C, a pH of 7.0, glucose as the carbon source, ammonium chloride as the nitrogen source, and a carbon-to-nitrogen ratio of 4:1, resulting in a 1.42-fold PHB production increase. B. pacificus was also cultured on various low-cost substrates. Among the oil wastes, feedstock showed the highest PHB production (1.983 ± 0.005g/L) and among agricultural residues, the maximum PHB was obtained from rice bran (1.626 ± 0.01g/L). UV-visible spectrophotometric, FT-IR, and HR-LCMS analysis of extracted PHB confirmed characteristics of PHB molecules (ʎ-max at 210nm, functional groups between 1152 and 2925cm-1). The 1H NMR analysis revealed distinct signals for protons resonating at aliphatic CH3 proton groups, bridged CH protons, and shielding CH2 proton regions that matched PHBs. Thermal gravimetric analysis (TGA) and direct scanning colorimetric (DSC) analysis revealed 89.4% degradation and melting temperature at 124.1°C for the extracted PHB compound.

Nutrient balance for enhanced recovery of stressed Spirulina platensis.

The failure of mass production of Spirulina plateaus can be attributed to an imbalance of nutrients (C:N) and an increase in accumulated sodium ions, coupled with the traditional harvesting process. The current study aims at the recovery of stressed and red cultures of Spirulina platensis as well as enhanced phycocyanin accumulation. The stressed Spirulina platensis cultures were obtained from a local Egyptian Spirulina production farms, which were further subjected to water analyses after removing the Spirulina biomass. Optimization was performed within 300-ml water path photobioreactor. Spirulina platensis samples were incubated with Zarrouk medium comparing with those modified using ammonium bicarbonate or ammonium acetate instead of sodium bicarbonate. Continuous batching was performed every 12days during three sequenced batches. Growth measurements (dry weight and pigments) were performed along the incubation time. It was found that carbon content of the growth medium seems to be more effective in Spirulina growth and biomass characteristics. Under different carbon sources, acetate resulted in the maximum dry weight of 1.48g·l-1 and recovery percentage of 463.3%. Such effect was extended along the different incubation batches. Various carbon concentrations revealed that moderate concentration of carbon in the form of acetate (0.699g·l-1) leads to the maximum growth under the same nitrogen content. A similar trend was observed with chlorophyll and phycocyanin accumulation, while carotenoids showed the opposite manner.

Potato and dairy industry side streams as feedstock for fungal and plant cell cultures

Utilization of industrial side streams as nutrient source for cellular agriculture is a promising option to improve the sustainability of the production processes. The aim of the study was to evaluate usability of two liquid food industry side streams, potato cell fluid and acid whey, as nutrient source for production of food ingredients with plant cell cultures (arctic bramble (Rubus arcticus L.) and tobacco BY-2 (Nicotiana tabacum L.)) and filamentous fungi (Paecilomyces variotii, Rhizopus oligosporus and Trichoderma reesei). The side streams were found to contain various sugars and other potential nutrients suitable for the studied organisms. The side streams were used as a sole nutrient source (fungi) or as a replacement of selected nutrients in the growth media (fungi and plant cells) in flask scale cultivations. Acid whey was successfully used as a growth medium for filamentous fungi, but it inhibited plant cell growth presumably due to high organic acid content (14 g/l). Potato side stream was found suitable as a fungal and plant cell growth media supplement, and it was used together with glucose for filamentous fungi, or as a partial replacer of macronutrients for plant cells, in bioreactor cultivations yielding high fungal (up to 36 g/l dry weight) and good plant cell (9.5 g/l dry weight) biomass production. Analyses of the produced biomasses revealed good nutritional value in terms of amino acid (17–27 % of dry matter) and dietary fiber (23–30 % of dry matter) contents, giving good premises for utilization in human nutrition.

The role of Bacillus strains and growth medium in shaping γ-glutamyl peptide production

To compare γ-glutamyl di- and tripeptides generated by different Bacillus strains and investigate the impact of substrates on the production of γ-glutamyl peptides, six strains of four species, including B. subtilis, B. velezensis, B. amyloliquefaciens and B. paralicheniformis, were cultivated in a standard brain heart infusion (BHI) broth and a medium consisting of hemoglobin hydrolysates (HH) for six days. Quantitative analysis of free amino acids and γ-glutamyl peptides, as well as determination of bacterial growth and γ-glutamyltransferase activity were carried out. Results revealed that all Bacillus strains could generate a series of γ-glutamyl dipeptides in both media. Most strains produced higher concentrations of target peptides in the HH medium (up to 83.56 μM), which correlated with higher levels of free amino acids in the medium. Glutathione was detected only in the BHI medium with B. subtilis PRO84, B. velezensis PRO76, B. altitudinis PRO107, and B. paralicheniformis PRO109 (up to 0.61 μM), indicating the glutathione-forming ability of these Bacillus strains. Production of γ-glutamyl peptides was influenced by both the choice of strain and growth medium, with the medium exerting a more pronounced impact than the strain. This study highlights the importance of selecting the appropriate substrate and Bacillus strains to produce desired γ-glutamyl peptides as enhancers of koku, potentially inspiring the conversion of meat side streams into valuable kokumi seasonings.

Biochemical and Molecular Identification of Azospirillum brasilense Bacteria and Evaluation of Their Efficiency in Producing Hormones, Dissolving Phosphorus, and Fixing Nitrogen

The study aimed to isolate A.brasilense bacteria from the soil of the rhizosphere of different plants and different locations in Al-Diwaniyah Governorate. They were identified in two ways. The first was the routine method, which included studying the microscopic and cultural characteristics and biochemical tests of the isolates. The second method was molecular, using polymerase chain reaction (PCR) technology and using primers. It also included testing the efficiency of these isolates in dissolving tricalcium phosphate (TCP) on Pikovskaya agar medium, fixing nitrogen in the liquid nutrient medium (N.B), and measuring the amount of hormone production using an HPLC device. The results of isolation and regular and molecular identification the presence of ten isolates of bacteria bearing the characteristics of A. brasilense bacteria, out of fifteen local bacterial isolates, took the following symbols and sequences (Az2, Az3, Az5, Az6, Az7, Az9, Az11, Az12, Az13, Az14), as the results showed confirmation of the identification of the bacterial isolates identified by biochemical tests. Using a specialized primer to amplify the 462bp fragment of the 16S ribosomal RNA gene, the results of testing the efficiency of the bacteria in dissolving phosphate (TCP) showed that the isolate (Az13) outperformed the highest value in its effectiveness in dissolving metallic phosphorus through the diameter of the clear zone around the colony, which was effective in dissolving phosphate of up to 3.89 mm. As for the nitrogen fixation efficiency test, the isolate (Az3) excelled in the amount of fixed nitrogen reaching 12.44 mg.L-1. As for the amount of its production of growth regulators for Auxins, Gibberellins, and Cytokinins, the isolate (Az3) recorded the highest value in the amount of Auxins production ( Indol-3-acetic acid) its concentration reached 34.4 µg.ml-1, and as for the production of the hormone Gibberellins (GA3), the isolate (Az3) recorded the highest value amounting to 34.7 µg.ml-1, as for the production of the hormone Cytokinins (CK), The isolate (Az11) recorded the highest value, amounting to 28.8 µg.ml-1.

Development of mixed linear models to analyze and describe the impact of malathion on the larval growth of Megaselia scalaris (Diptera: Phoridae) under various feeding media and environmental conditions.

Forensic entomology plays a crucial role in estimating the minimum postmortem interval through the study of insect larvae found at crime scenes. The precision of this estimation relies on various biotic and abiotic elements that simultaneously influence insect growth and development, encompassing factors such as temperature, humidity, photoperiod, diet, and the existence of xenobiotics in decomposing tissues. Despite numerous studies on the influence of these factors, including the impact of xenobiotics, there are currently no robust tools available for making corrections to this estimation considering concurrently all variables. In an attempt to propose an exploratory and descriptive statistical model to analyze the simultaneous effect and interaction of different variables on larval growth, this study aimed to compare the effect of malathion on the growth of Megaselia scalaris (Loew, 1866) (Diptera: Phoridae) raised in malathion-spiked porcine muscle, under controlled and uncontrolled temperature and humidity conditions (environmental conditions). Larvae were also reared using various growth media. A split-plot design that combined crossed and nested factors was employed; 2 linear mixed models were developed to assess the relationships between the variables. The model provides valuable insights into the complex interactions among xenobiotics, growth media, and environmental conditions in the size and development of M. scalaris.

Determining Rates of Molecular Secretion from Supernatant Concentration Measurements in a 3D-Bioprinted Human Liver Tissue Model.

The secretion rate of albumin is a key indicator of function in liver tissue models used for hepatotoxicity and pharmacokinetic testing. However, it is not generally clear how to determine molecular secretion rates from measurements of the molecular concentration in supernatant media. Here, we develop computational and analytical models of molecular transport in an experimental system that enable determination of albumin secretion rates based on measurements of albumin concentration in supernatant media. The experimental system is a 3D-bioprinted human liver tissue construct embedded in a 3D culture environment made from packed microgel particles swollen in liquid growth media. The mathematical models reveal that the range of albumin synthesis rates necessary to match experimentally measured albumin concentrations corresponds to reaction-limited conditions, where a steady state of albumin spatial distribution is rapidly reached between media exchanges. Our results show that temporally resolved synthesis rates can be inferred from serial concentration measurements of collected supernatant media. This link is critical to confidently assessing in vitro tissue performance in applications where critical quality attributes must be quantified, like in drug development and screening.