Mixed Culture Research Articles

Feasibility study on heterotrophic utilization of galactose by Chlorella sorokiniana and promotion of galactose utilization through mixed carbon sources culture

BackgroundThe development of alternative carbon sources is important for reducing the cost of heterotrophic microalgae cultivation. Among cheap feedstocks, galactose is one of the most abundant sugars and can be easily obtained from many natural biomasses. However, it is generally difficult to be utilized by microalgae. In addition, the mechanism of its low utilization efficiency in heterotrophic cultivation is still unknown.ResultsAmong seven tested carbon sources, only glucose and acetate could be efficiently utilized by C. sorokiniana in heterotrophic cultivation while there were no apparent signs of utilization of other carbohydrates, including galactose, in regular heterotrophic cultivation. However, galactose could be utilized in cultures with high inoculation sizes. This confirmed that C. sorokiniana has a complete pathway for transporting and assimilating galactose under dark conditions, but the rate of galactose utilization is quite low. In addition, the galactose utilization was greatly enhanced in mixotrophic cultures, which indicated that galactose utilization could be enhanced by additional pathways that can enhance cell growth. Based on above results, a mixed carbon source culture strategy was proposed to improve the utilization rate of galactose, and a significant synergistic effect on cell growth was achieved in cultures using a mixture of galactose and acetate.ConclusionsThis study indicated that the galactose metabolism pathway may not be inherently deficient in Chlorophyta. However, its utilization rate was too low to be detected in regular heterotrophic cultivation. Mixed carbon source culture strategy was confirmed effective to improve the utilization rate of galactose. This study contributes to a deeper understanding of the utilization ability of difficultly utilized substrates in the heterotrophic cultivation of microalgae, which is of great significance for reducing the cost of heterotrophic cultivation of microalgae.

Phycoremediation of Potato Industry Wastewater for Nutrient Recovery, Pollution Reduction, and Biofertilizer Production for Greenhouse Cultivation of Lettuce and Celery in Sandy Soils

Microalgae-based wastewater treatment offers an eco-friendly opportunity for simultaneous nutrient recovery and biomass generation, aligning with the circular bioeconomy concept. This approach aims to utilize the nutrients of potato industry wastewater (PIW) for algal growth while mitigating the environmental impact of this industrial byproduct. This study focused on cultivating three cyanobacterial strains, Anabaena oryzae, Nostoc muscorum, and Spirulina platensis, in PIW and synthetic media for 30 days to assess feasibility. Growth performance was monitored by measuring chlorophyll content, dry weight (DW), optical density (OD), and pH at 3-day intervals. The high-performing cyanobacterial biomass from the laboratory findings was formulated into a biofertilizer, which was then evaluated in a controlled greenhouse experiment on celery and lettuce plants. The biofertilizer replaced conventional NPK mineral fertilizers at different levels (25%, 50%, and 75%), while a control group received 100% chemical fertilizer. The results showed favourable growth of all three cyanobacteria strains and their mixture in PIW throughout the experiment. The mixed cyanobacteria followed by Spirulina platensis exhibited the highest growth rates, achieving chlorophyll contents of 3.75 and 2.30 µg·mL−1, DWs of 1.79 g·L−1 and 1.63 g·L−1, and ODs of 0.41 and 0.38, respectively, surpassing the other treatments. The formulated biofertilizers, Spi-PIW (Spirulina platensis + potato industry wastewater) and Cyano-PIW (mixed culture+ potato industry wastewater), significantly enhanced plant height, root and stem lengths, and the number of leaves per plant in celery and lettuce compared to the control group. These biofertilizer treatments also improved chlorophyll contents, as well as macro- and micronutrient levels, in the two crops. Additionally, the application of these biofertilizers improved certain sandy soil properties, i.e., pH, total organic matter, total nitrogen, phosphorus, and potassium. In conclusion, utilizing PIW as a substrate for cultivating cyanobacteria strains and producing high-quality liquid bio-organic fertilizers holds potential for reducing recommended NPK fertilizer doses by 25–50% in celery and lettuce growth, providing an environmentally friendly approach.

Optimization of methane gas-liquid mass transfer during biogas-based ectoine production in bubble column bioreactors

Nowadays, the utilization of biogas for energy generation is hindered by the declining production costs of solar and wind power. A shift towards the valorization of biogas into ectoine, a highly valuable bioproduct priced at 1000 €⸱kg−1, offers a novel approach to fostering a more competitive biogas market while contributing to carbon neutrality. This study evaluated the optimization of CH4 gas-liquid mass transfer in 10 L bubble column bioreactors for CH4 conversion into ectoine and hydroxyectoine using a mixed methanotrophic culture. The influence of the empty bed residence time (EBRTs of 27, 54, and 104 min) at different membrane diffuser pore sizes (0.3 and 0.6 mm) was investigated. Despite achieving CH4 elimination capacities (CH4-ECs) of 10–12 g⸱m−3⸱h−1, an EBRT of 104 min mediated CH4 limitation within the cultivation broth, resulting in a negligible biomass growth. Reducing the EBRT to 54 min entailed CH4-ECs of 21–24 g⸱m−3⸱h−1, concomitant to a significant increase in biomass growth (up to 0.17 g⸱L⸱d−1) and reaching maximum ectoine and hydroxyectoine accumulation of 79 and 13 mg⸱gVSS−1, respectively. Conversely, process operation at an EBRT of 27 min lead to microbial inhibition, resulting in a reduced biomass growth of 0.09 g⸱L⸱d−1 and an ectoine content of 47 mg⸱gVSS−1. While the influence of diffuser pore size was less pronounced compared to EBRT, the optimal process performance was observed with a diffuser pore size of 0.6 mm.

Isolation methods and characterization of primary rat neurovascular cells

BackgroundThere is significant interest in isolating cells of the blood-brain barrier (BBB) for use in in vitro screening of therapeutics and analyzing cell specific roles in neurovascular pathology. Primary brain cells play an advantageous role in BBB models; however, isolation procedures often do not produce cells at high enough yields for experiments. In addition, although numerous reports provide primary cell isolation methods, the field is lacking in documentation and detail of expected morphological changes that occur throughout culturing and there are minimal troubleshooting resources. Here, we present simplified, robust, and reproducible methodology for isolating astrocytes, pericytes, and endothelial cells, and demonstrate several morphological benchmarks for each cell type throughout the process and culture timeframe. We also analyze common considerations for developing neurovascular cell isolation procedures and recommend solutions for troubleshooting.ResultsThe presented methodology isolated astrocytes, pericytes, and endothelial cells and enabled cell attachment, maturation, and cell viability. We characterized milestones in cell maturation over 12 days in culture, a common timeline for applications of these cell types in BBB models. Phase contrast microscopy was used to show initial cell plating, attachment, and daily growth of isolated cells. Confocal microscopy images were analyzed to determine the identity of cell types and changes to cell morphology. Nuclear staining was also used to show the viability and proliferation of glial cells at four time points. Astrocyte branches became numerous and complex with increased culture time. Microglia, oligodendrocytes, and neurons were present in mixed glial cultures for 12 days, though the percentage of microglia and neurons expectedly decreased after passaging, with microglia demonstrating a less branched morphology.ConclusionsNeurovascular cells can be isolated through our optimized protocols that minimize cell loss and encourage the adhesion and proliferation of isolated cells. By identifying timepoints of viable glia and neurons within an astrocyte-dominant mixed culture, these cells can be used to evaluate drug targeting, uptake studies, and response to pathological stimulus in the neurovascular unit.

Metabolic picture of microbial interaction: chemical crosstalk during co-cultivation between three dominant genera of bacteria and fungi in medicinal plants rhizosphere.

Microbial communities affect several aspects of the earth's ecosystem through their metabolic interaction. The dynamics of this interaction emerge from complex multilevel networks of crosstalk. Elucidation of this interaction could help us to maintain the balance for a sustainable future. To investigate the chemical language among highly abundant microbial genera in the rhizospheres of medicinal plants based on the metabolomic analysis at the interaction level. Coculturing experiments involving three microbial species: Aspergillus (A), Trichoderma (T), and Bacillus (B), representing fungi (A, T) and bacteria (B), respectively. These experiments encompassed various interaction levels, including dual cultures (AB, AT, TB) and triple cultures (ATB). Metabolic profiling by LC-QTOFMS revealed the effect of interaction level on the productivity and diversity of microbial specialized metabolites. The ATB interaction had the richest profile, while the bacterial profile in the monoculture condition had the lowest. Two native compounds of the Aspergillus genus, aspergillic acid and the dipeptide asperopiperazine B, exhibited decreased levels in the presence of the AT interaction and were undetectable in the presence of bacteria during the interaction. Trichodermarin N and Trichodermatide D isolated from Trichoderma species exclusively detected during coexistence with bacteria (TB and ATB). These findings indicate that the presence of Bacillus activates cryptic biosynthetic gene clusters in Trichoderma. The antibacterial activity of mixed culture extracts was stronger than that of the monoculture extracts. The TB extract exhibited strong antifungal activity compared to the monoculture extract and other mixed culture treatments. The elucidation of medicinal plant microbiome interaction chemistry and its effect on the environment will also be of great interest in the context of medicinal plant health Additionally, it sheds light on the content of bioactive constituents,and facilitating the discovery of novel antimicrobials.

Clash of mosquito wings: Larval interspecific competition among the mosquitoes, Culex pipiens, Aedes albopictus and Aedes aegypti reveals complex population dynamics in shared habitats.

Globalisation, climate change and international trade are the factors contributing to the spread of Aedes albopictus (Diptera: Culicidae) and Ae. aegypti into new areas. In newly invaded habitats, these non-native species can serve as arbovirus disease vectors or increase the risk of disease spill over. These mosquitoes continue to emerge in new areas where they have or will have overlapping ranges with other resident mosquito species. The study investigates how invasive Aedes mosquitoes compete with the native Culex pipiens in Türkiye, which might affect the overall mosquito population dynamics and disease transmission risks. Both Aedes species exhibited contrasting responses to interspecific competition with Cx. pipiens. While Ae. albopictus suffers reduced emergence primarily in larger containers with abundant food, Ae. aegypti surprisingly thrives in mixed cultures under all food conditions. Adult Cx. pipiens emergence drops by half against Ae. albopictus and under specific conditions with Ae. aegypti. Competition influences mosquito size differently across species and life stages. Culex pipiens females grow larger when competing with Ae. aegypti, potentially indicating resource advantage or compensatory strategies. However, Ae. albopictus size shows more nuanced responses, suggesting complex interactions at play. Understanding how invasive and native mosquitoes interact with each other can provide insights into how they adapt and coexist in shared habitats. This knowledge can inform effective control strategies. The study highlights the differential responses of invasive Aedes species and the potential for managing populations based on their competitive interactions with the native Cx. pipiens. It can contribute to improved monitoring and prediction systems for the spread of invasive mosquitoes and the associated disease risks.

Eta-secretase-like processing of the amyloid precursor protein (APP) by the rhomboid protease RHBDL4

The amyloid precursor protein (APP) is a key protein in Alzheimer’s disease synthesized in the endoplasmic reticulum (ER) and translocated to the plasma membrane where it undergoes proteolytic cleavages by several proteases. Conversely to other known proteases, we previously elucidated rhomboid protease RHBDL4 as a novel APP processing enzyme where several cleavages likely occur already in the ER. Interestingly, the pattern of RHBDL4-derived large APP C-terminal fragments resemble those generated by the η-secretase or MT5-MMP, which was described to generate so called Aη fragments. The similarity in large APP C-terminal fragments between both proteases raised the question whether RHBDL4 may contribute to η-secretase activity and Aη-like fragments. Here, we identified two cleavage sites of RHBDL4 in APP by mass spectrometry, which, intriguingly, lie in close proximity to the MT5-MMP cleavage sites. Indeed, we observed that RHBDL4 generates Aη-like fragments in vitro without contributions of α-, β-, or γ-secretases. Such Aη-like fragments are likely generated in the ER since RHBDL4-derived APP-C-terminal fragments do not reach the cell surface. Inherited, familial APP mutations appear to not affect this processing pathway. In RHBDL4 knockout mice, we observed increased cerebral full length APP in comparison to wild type (WT) in support of RHBDL4 being a physiologically relevant protease for APP. Furthermore, we found secreted Aη fragments in dissociated mixed cortical cultures from WT mice, however significantly less Aη fragments in RHBDL4 knockout cultures. Our data underscores that RHBDL4 contributes to η-secretease-like processing of APP and that RHBDL4 is a physiologically relevant protease for APP.

Crustacean endocrinology: Sexual differentiation and potential application for aquaculture

Crustaceans, which represent a significant subset of arthropods, are classified into three major classes: Ostracoda, Malacostraca, and Branchiopoda. Among them, sex manipulation in decapod species from the Malacostraca class has been extensively researched for aquaculture purposes and to study reproductive physiology and sexual plasticity. Some decapods exhibit sexual dimorphism that influences their biological and economic value. Monosex culture, in which only one sex is cultivated, increases production yields while reducing the risk of invasiveness, as genetic leakage into natural waters is less likely to occur. Differences in yield are also observed when cultivating different sexes, with all-male cultures of Macrobrachium rosenbergii being more profitable than both mixed and all-female cultures. Research on decapod sexual differentiation has led to a better understanding of sex determination and sexual differentiation processes in arthropods. Similar to most mammals and other vertebrate classes, Malacostraca crustaceans, including decapods, exhibit a cell-non-autonomous mode of sexual development. Genetic factors (e.g., sex chromosomes) and endocrine factors (e.g., insulin-like androgenic gland factor and crustacean female sex hormone) play pivotal roles in the development of sexually dimorphic traits. This review synthesizes the existing understanding of sex determination mechanisms and the role of sex hormones in decapod species. Additionally, it provides an overview of the methyl farnesoate, which has been suggested to be involved in male sex differentiation in some crab species, as well as the phenomenon of male-to-female sex reversal in host decapods caused by parasitic crustaceans.

Development of Salmonella enterica serovar Typhimurium, Listeria monocytogenes, and Pseudomonas aeruginosa multi-species in vitro dry surface biofilm models: Insights into resilience and persistence in low-moisture environments

Salmonella enterica serovar Typhimurium, Listeria monocytogenes, and Pseudomonas aeruginosa can persist as dry surface biofilms (DSB) and survive desiccation for prolonged periods of time. This could serve as potential sources of contamination in low-moisture food (LMF) processing environments. The objectives of this study were to develop in vitro mono-and mix-culture DSB models to better understand pathogen survival under dehydrated conditions as DSB. Mono-and mix-culture wet surface biofilms (WSB) of S. enterica ser. Typhimurium ATCC 14028, L. monocytogenes FSL R8-5318, and P. aeruginosa ATCC 15442 were developed following EPA MLB SOP MB-19 protocol on borosilicate glass coupons using a CDC® biofilm reactor. WSB were harvested and dried for 24, 48, and 72 h at 21 °C to form DSB. Mean log10 CFU/cm2 were calculated at each desiccation time point and compared to control (WSB) using LSM of PROC GLM to fit linear models (P = 0.05). Approximately 8.8 ± 1.7, 7.1 ± 1.1, 6.9 ± 0.6, and 6.3 ± 0.8 mean log10 CFU/coupon were recovered from mono-culture Salmonella DSB at 0, 24, 48, and 72 h, respectively. Overall, Salmonella mean log10 densities were limited when co-cultured with P. aeruginosa although there were no significant differences in Salmonella density over desiccation time points. Mean log10 CFU/coupon for L. monocytogenes monoculture DSB were comparatively lower i.e., 5.7 ± 0.4 (0 h), 3.8 ± 0.1 (24 h), and 3.4 ± 0.6 (48 h), and 2.6 ± 0.3 (72 h). However, when co-cultured with P. aeruginosa, L. monocytogenes exhibited consistently higher cell densities 6.7 ± 0.4, 6.0 ± 0.9, 5.6 ± 0.04, and 4.9 ± 0.6, at 0, 24, 48, and 72 h, respectively. On average, P. aeruginosa consistently exceeded six log10 at each desiccation time point in co-culture. Confocal laser scanning microscopy (CLSM) images confirmed biofilm development and persistence over time in mono and mixed cultures. This work provides foundational evidence toward better understanding pathogen survival under dehydrated conditions, interactions between different pathogens within desiccated biofilms, and their respective survival rates. Additionally, these data provide quantitative assessment of bacterial load within DSB under high-risk scenarios for LMFs. These models provide opportunities to explore targeted mitigation strategies, including efficacy of dry sanitation practices, to prevent the formation or remove DSB in LMF processing environments.

Inhibition of Sphingosine Kinase 1 Reduces Sphingosine-1-Phosphate and Exacerbates Amyloid-Beta-Induced Neuronal Cell Death in Mixed-Glial-Cell Culture.

In Alzheimer's disease (AD) pathology, the accumulation of amyloid-beta (Aβ), a main component of senile plaques, activates glial cells and causes neuroinflammation. Excessive neuroinflammation results in neuronal dropouts and finally produces the symptoms of AD. Recent studies suggest that disorder in sphingosine-1-phosphate (S1P) metabolism, especially the decreased expression of sphingosine kinase (SK)1, followed by the reduction in the amount of S1P, can be a promotive factor in AD onset. Thus, we explored the possibility that dysregulated S1P metabolism affects AD through the altered function in glial cells. We evaluated the effect of PF-543, a pharmacological inhibitor of SK1, on the inflammatory responses by lipopolysaccharide (LPS)-activated glial cells, microglia, and astrocytes. The treatment with PF-543 decreased the intracellular S1P content in glial cells. The PF-543 treatment enhanced the nitric oxide (NO) production in the LPS-treated neuron/glia mixed culture. Furthermore, we found that the augmented production of NO and reactive oxygen species (ROS) in the PF-543-treated astrocytes affected the microglial inflammatory responses through humoral factors in the experiment using an astrocyte-conditioned medium. The PF-543 treatment also decreased the microglial Aβ uptake and increased the number of injured neurons in the Aβ-treated neuron/glia mixed culture. These results suggest that a decrease in the glial S1P content can exacerbate neuroinflammation and neurodegeneration through altered glial cell functions.

Effect of Storage on Probiotic Viability, Physicochemical and Sensory Properties of Probiotic-enriched Orange Juice

The effect of enrichment of orange juice with probiotic Lactobacillus plantarum and Lactobacillus bulgaricus as single and mixed cultures on the cell viability, physicochemical parameters and sensory properties of the juice during a 20day refrigerated (4oC) and room temperature (30± 2oC) conditions was studied. The viable cell count remained relatively stable at 4oC over the storage period for both strains but was at its peak after 10 days of storage at 30±2oC with L. bulgaricus accounting for the highest count (9.36±0.04 CFU/mL). The acidity of the enriched juice increased as the storage period progressed at 30oC with the highest acidity observed with L. bulgaricus (pH 3.00±0.02, titratable acidity 16.27±0.05mg lactic acid/mL). Sensory evaluation indicated that the juice enriched with L. bulgaricus was more acceptable. The study concluded that orange juice enriched with L. bulgaricus is suitable in the development of non-dairy based functional food and could act as an ideal food matrix for probiotic beverage.

A modified mycobacterial growth inhibition assay for the functional assessment of vaccine-mediated immunity

The Mycobacterial growth inhibition assay (MGIA) is an ex-vivo assay used to measure the overall functional immune response elicited by infection or vaccination. In tuberculosis (TB) vaccine development, MGIA is a potentially important tool for preclinical evaluation of early-stage vaccine candidates to complement existing assays, and to potentially reduce the need for lengthy and costly pathogenic Mycobacterium tuberculosis (Mtb) animal challenge experiments. The conventional method of MGIA in mice entails directly infecting mixed cell cultures, most commonly splenocytes, from immunised mice with mycobacteria. However, this direct infection of mixed cell populations may yield unreliable results and lacks sufficient sensitivity to discriminate well between different vaccines due to the low number of mycobacteria-permissive cells. Here, we modified the assay by inclusion of mycobacteria-infected congenic murine macrophage cell lines as the target cells, and by measuring the total number of killed cells rather than the relative reduction between different groups. Thus, using splenocytes from Mycobacterium bovis BCG immunised mice, and J774 and MH-S (BALB/c background) or BL/6-M (C57Bl/6 background) macrophage cell lines, we demonstrated that the modified assay resulted in at least 26-fold greater mycobacterial killing per set quantity of splenocytes as compared to the conventional method. This increased sensitivity of measuring mycobacterial killing was confirmed using both the standard culture forming unit (CFU) assay and luminescence readings of luciferase-tagged virulent and avirulent mycobacteria. We propose that the modified MGIA can be used as a highly calibrated tool for quantitating the killing capacity of immune cells in preclinical evaluation of vaccine candidates for TB.

Collaboration of bacterial consortia for biodegradation of high concentration phenol and potential application of machine learning

Mixed culture of microorganisms is an effective method to remove high concentration of phenol in wastewater. At present, it is still a challenge for microorganisms to remove high-concentration phenol from wastewater. In this study, a phenol-degrading consortium was isolated, which could rapidly degrade 1800 mg/L phenol within 30 h, and the highest phenol degradation concentration was 2000 mg/L. Further exploration of how microbial consortium cooperates to promote phenol biodegradation was studied: the core bacteria of the microbial consortium was relatively stable during phenol degradation; the bacteria could improve the adaptability to environment and metabolic ability of phenol, by producing more surfactants and betaine, thereby improving the degradation rate. The determination coefficient (R2) in the machine learning model showed that the back propagation artificial neural network (BP-ANN) can predict the biodegradation of phenol under different conditions, saving time and economic costs. This study explains how microbial consortium cooperates to degrade phenol from the aspects of microbial consortium composition and metabolic analysis, which provides a theoretical basis for mixed culture microorganisms to degrade pollutants.

Performance of salt‐bridge microbial fuel cell (SB‐MFC) with various microorganism cultures on the generation of electricity from tofu wastewater

A suitable wastewater treatment system is required due to the high organic compound content in tofu wastewater, which can harm the environment. Biological treatment methods are effective for treating tofu wastewater due to its characteristics. Microbial fuel cells (MFCs) represent one such biological treatment option, effectively removing organic contaminants while generating low‐power electricity through bioenergetic reactions. In MFCs, microorganisms are used as biocatalysts to degrade the organic compounds present in wastewater. This study aimed to assess the efficacy of Salt‐bridge microbial fuel cells (SB‐MFC) using various acclimatized microbe cultures for reducing organic compounds and generating energy from tofu wastewater. Tofu wastewater was sterilized prior to introduction into the reactor. Additional microbes, including the native microbe consortium from tofu wastewater, Escherichia coli, Saccharomycopsis fibuligera, and a mixed culture of E. coli and S. fibuligera, were then introduced as biocatalysts. Carbon electrodes were utilized as both the anode and cathode. The results indicate that the mixed culture of E. coli and S. fibuligera significantly reduced COD and BOD5 levels, with removal rates of 82.74% and 76.53%, respectively, after 48 h. Furthermore, the culture generated a voltage of 676 mV, a current of 2.53 mA, a power density of 428 mWatt/m2, and 4.789×10‐2 kWh of energy. This study contributes to the advancement of SB‐MFC by utilizing wastewater and a combination of bacteria and yeast as biocatalysts.

Anaerobic Treatment of N-(Phosphonometyl) Glycine Using Mixed Culture in Batch Reactor

Pesticides have been used in the fight against various pests to increase crop growth in our country and the world. In recent years, high concentrations of toxic and detection of carcinogenic pesticides indicates that these substances are transported to groundwater and pollute water resources. N- (phosphonomethyl) glycine are also used in large quantities. In this study, the optimization of anaerobic treatment of N- (phosphonomethyl) glycine was investigated by applying a statistical-based experimental design. Full factorial trials different initial pesticide concentrations and cosubstrate type were established, making up 9 different experimental setups. Trials were performed in 2 replicates. The experiments were carried out in Oxitop C bottles with stirring at a working volume of 200 mL. The pH was adjusted to 7 ± 0.2. Trials were performed at 35oC for 30 days. At the end of the process, the removal of the trials according to the input and output COD and pesticide values were examined. As a result, the most efficient COD removal was achieved 98,96 % when pesticide concentration of 5 mg L-1 and glucose as cosubstrate were used. The highest pesticide removal was found to be 74,83% when pesticide concentration of 25 mg L-1 and glucose was used as cosubstrate.

Anti-Leukemic Effects Induced by Dendritic Cells of Leukemic Origin from Leukemic Blood Samples Are Comparable under Hypoxic vs. Normoxic Conditions.

Hypoxia can modulate the immune system by affecting the function and activity of immune cells, potentially leading to altered immune responses. This study investigated the generation of leukemia-derived dendritic cells (DCleu) from leukemic blasts and their impact on immune cell activation under hypoxic (5-10% O2) compared to normoxic (21% O2) conditions using various immunomodulatory Kits. The results revealed that DC/DCleu-generation was similar under hypoxic and normoxic conditions, with no significant differences observed in frequencies of generated DC/DCleu. Furthermore, the study showed that the activation of immune cells and their anti-leukemic activity improved when T cell-enriched immunoreactive cells were co-cultured with DC/DCleu which were generated with Kit I and M compared to the control after mixed lymphocyte cultures. The anti-leukemic activity was improved under hypoxic compared to normoxic conditions after MLCWB-DC Kit M. These findings suggest that DC/DCleu-cultures of leukemic whole blood with Kits under hypoxic conditions yield comparable frequencies of DC/DCleu and can even increase the anti-leukemic activity compared to normoxic conditions. Overall, this research highlights the potential of utilizing DC/DCleu (potentially induced in vivo with Kits) as a promising approach to enhance immune response in patients with acute myeloid leukemia.

Hybrid pathway of bio-ethanol production employing empty fruit bunches co-gasified with charcoal and mixed culture fermentation: Optimization using response surface methodology

The research focuses on the synthesis of syngas from lignocellulosic biomass via co-gasification, employing response surface methodology. In addition, the study aims to optimize syngas fermentation efficiency for bioethanol production, utilizing two species of microorganisms – Saccharomyces cerevisiae and Clostridium butyricum – within an integrated biorefinery framework. The raw syngas was generated through the co-gasification of empty fruit bunches and charcoal mixtures (75:25). The optimal ratio of these components was determined via central composite design. The individual performances of S. cerevisiae and C. butyricum, as well as their co-fermentation, were thoroughly investigated, considering parameters such as colony forming units (CFU), pH, total organic carbon (TOC), and syngas flow rate. Morphological features of microorganisms during syngas fermentation were characterized using field emission scanning electron microscopy analysis. The results indicated that the highest bioethanol concentration, reaching 31.20 mmol, was achieved through co-fermentation as opposed to single-inoculum fermentation. Furthermore, a significant increase (3.08%) in bioethanol productivity was observed when the syngas flow rate was elevated from 50 to 1000 mL/min. Therefore, microbial co-fermentation emerges as a promising strategy to enhance bioethanol production from syngas derived from lignocellulosic biomass. Highlights Parametric optimization for co-gasification using response surface methodology Syngas was co-fermented using Saccharomyces cerevisiae and Clostridium butyricum Effects of syngas (CFU, pH, and TOC) were investigated during co-fermentation Bioethanol production increased 3.08% when syngas flowrate increased from (50 to 1000 mL/min)

Compound-Specific Carbon, Nitrogen, and Hydrogen Isotope Analysis to Characterize Aerobic Biodegradation of 2,3-Dichloroaniline by a Mixed Enrichment Culture.

Compound-specific isotope analysis (CSIA) is an established tool to track the in situ transformation of organic chemicals at contaminated sites. In this work, we evaluated the potential of multi-element CSIA to assess biodegradation of 2,3-dichloroaniline (2,3-DCA), which is a major industrial feedstock. Using controlled laboratory experiments, we determined, for the first time, negligible carbon (<0.5‰) and hydrogen (<10‰) isotope fractionation and a significant inverse nitrogen isotope fractionation (>10‰) during aerobic 2,3-DCA biodegradation by a mixed enrichment culture. The tentative identification of a glutamate conjugate of 2,3-DCA as a reaction intermediate indicates that the initial multistep enzymatic reaction may be rate-limiting. The formation of the glutamate adduct would increase the bond energy at the N atom, thus likely explaining the observed inverse N isotope fractionation. The corresponding nitrogen enrichment factor was +6.8 ± 0.6‰. This value was applied to investigate the in situ 2,3-DCA biodegradation at a contaminated site where the carbon and nitrogen isotope signatures from field samples suggested similar aerobic processes by native microorganisms. Under the assumption of the applicability of the Rayleigh model in a pilot wetland treating contaminated groundwater, the extent of biodegradation was estimated to be up to 80-90%. This study proposes multi-element CSIA as a novel application to study 2,3-DCA fate in groundwater and surface water and provides insights into biodegradation pathways.

&lt;span&gt;&lt;strong&gt;Enhanced thiamethoxam degradation in water&amp;nbsp; and soil by &lt;/strong&gt;&lt;em&gt;Phanerochaete &lt;/em&gt;&lt;strong&gt;sp. Th1 and &lt;/strong&gt;&lt;em&gt;Ensifer &lt;/em&gt;&lt;strong&gt;sp. Th2&lt;/strong&gt;&lt;/span&gt;

Thiamethoxam is widely applied to control pest insects, but it causes serious environmental contamination. In this study, the degradation processes of the compound in soil and water collected from a rice field by native microorganisms, by mixed culture of Phanerochaete sp. Th1 and Ensifer sp. Th2, and by both the mixed pure culture and native microorganisms were conducted. The result showed that the mixed culture inoculation increased thiamethoxam in all media. In liquid media, the degradation percentage of the substrate in collected water was 21.8±4.4%, collected water + inoculation was 44.2±5.0%, and mineral medium + inoculation was 98.0±0.4%. Degradation determination in media with 70% collected water and 30% dry soil showed that the main degradation occurred in the liquid phase. The degradation rates in the media with 50% collected water and 50% dry soil were higher in the surface layer, and lower in the bottom. Indigenous microorganisms also played an important role in the degradation process. This study provided valuable information on the thiamethoxam degradation in simulated media of a rice field and the role of Phanerochaete sp. Th1 and Ensifer sp. Th2 to enhance the degradation.

Effects of Inoculating Autochthonous Starter Cultures on Changes of N-Nitrosamines and Their Precursors in Chinese Traditional Fermented Fish during In Vitro Human Digestion.

The objective of this research was to investigate the impact of inoculating autochthonous starter cultures on the alterations in microorganisms, biogenic amines, nitrite, and N-nitrosamines in Chinese traditional fermented fish products (CTFPs) during in vitro human digestion. The results revealed that gastric digestion significantly (p < 0.05) inhibited the proliferation of lactic acid bacteria, yeast, Staphylococcus, and Enterobacteriaceae, whereas various microorganisms proliferated extensively during small intestine digestion. Meanwhile, small intestine digestion could significantly increase (p < 0.05) levels of putrescine, cadaverine, and tyramine. The reduced content observed in inoculated fermentation groups suggests that starter cultures may have the ability to deplete biogenic amines in this digestion stage. Gastric digestion significantly (p < 0.05) inhibited nitrite accumulation in all CTFPs samples. Conversely, the nitrite content increased significantly (p < 0.05) in all groups during subsequent small intestine digestion. However, the rise in the inoculated fermentation groups was smaller than that observed in the spontaneous fermentation group, indicating a potentially positive role of inoculated fermentation in inhibiting nitrite accumulation during this phase. Additionally, gastric digestion significantly (p < 0.05) elevated the levels of N-nitrosodimethylamine (NDMA) and N-nitrosopiperidine in CTFPs. Inoculation with L. plantarum 120, S. cerevisiae 2018, and mixed starter cultures (L. plantarum 120, S. cerevisiae 2018, and S. xylosus 135 [1:1:1]) effectively increased the degree of depletion of NDMA during this digestion process.