This study aimed to explore the fermentation behavior of selected potential probiotic strains during sorghum fermentation, considering their microbial counts, pH levels, and antibacterial activity, their ability to break down starch. The fermentation processes were conducted in the presence and absence of glucose, and the lactic acid bacterial strains underwent controlled fermentation to assess microbial counts, amylase activity, pH, and lactic acid levels in the presence and absence of glucose. In addition, the inhibitory activity of these potential probiotic strains against pathogenic bacteria (Salmonella typhimurium, Escherichia coli, and Staph-ylococcus aureus) was examined through co-culturing. The results demonstrated that all nine Lactobacillus strains exhibited robust growth in both sugar and sugar-free fermentation experiments. Glucose supple-mentation significantly enhanced lactic acid yield which was in a range of 0.19-0.52 % compared to fer-mentation without glucose with a range of 0.03-0.29 %. The selected Lactobacillus strains effectively lowered the pH to less than 4 after 24 hours, producing substantial lactic acid. Notably, in the absence of glucose, only Lb. helveticus D7 and Lb. amylolyticus D12 achieved pH levels below 4 after 8 hours, producing the highest lactic acid amounts of 0.27 and 0.29 % after 24 hours, respectively. Amylase activity was detected on two strains D7 and D12. Furthermore, most of the tested Lactobacillus strains demonstrated complete inhibition of pathogen growth after 24 hours of co-culturing, suggesting their potential for enhancing the safety quality of sorghum-based fermented products.

Aiming at the limitations of single-functionality, limited applicability, and complex designs prevalent in current metasurfaces, we propose a terahertz multifunctional and multiband tunable metasurface utilizing a VO2-metal hybrid structure. This metasurface structure comprises a top VO2-metal resonance layer, a middle polyimide dielectric layer, and a gold film reflective layer at the bottom. This metasurface exhibits multifunctionality, operating independently of polarization and incident angle. The varying conductivity states of the VO2 layers, enable the metasurface to achieve various terahertz functionalities, including single-band absorption, broadband THz absorption, and multiband perfect polarization conversion for linear (LP) and circularly polarized (CP) incident waves. Finally, we believe that the functional adaptability of the proposed metasurface expands the repertoire of options available for future terahertz device designs.

Pozol is a Mexican beverage prepared from fermented nixtamalized maize dough. To contribute to understanding its complex microbial ecology, the effect of inoculating on MRS-starch pure and mixed cultures of amylolytic Sii-25124 and non-amylolytic W. confusa 17, isolated from pozol, were studied on their interactions and fermentation parameters. These were compared with L. plantarum A6, an amylolytic strain isolated from cassava. Microbial growth, kinetic parameters, amylolytic activity, lactic acid production, and hydrolysis products from starch fermentation were measured. The population dynamics were followed by qPCR. L. plantarum A6 showed higher enzymatic activity, lactic acid, biomass production, and kinetic parameters than pozol LAB in pure cultures. Mixed culture of each pozol LAB with L. plantarum A6 showed a significant decrease in amylolytic activity, lactic acid yield, specific growth rate, and specific rate of amylase production. The interaction between Sii-25124 and W. confusa 17 increased the global maximum specific growth rate (µ), the lactic acid yield from starch (Ylac/s), lactic acid yield from biomass (Ylac/x), and specific rate of lactic acid production (qlac) by 15, 30, 30, and 40%, respectively compared with the pure culture of Sii-25124. Interactions between the two strains are essential for this fermentation.

The search for novel brewing strains from non-brewing environments represents an emerging trend to increase genetic and phenotypic diversities in brewing yeast culture collections. Another valuable tool is hybridization, where beneficial traits of individual strains are combined in a single organism. This has been used successfully to create de novo hybrids from parental brewing strains by mimicking natural Saccharomyces cerevisiae ale x Saccharomyces eubayanus lager yeast hybrids. Here, we integrated both these approaches to create synthetic hybrids for lager fermentation using parental strains from niches other than beer. Using a phenotype-centered strategy, S. cerevisiae sourdough strains and the S. eubayanus x Saccharomyces uvarum strain NBRC1948 (also referred to as Saccharomyces bayanus) were chosen for their brewing aptitudes. We demonstrated that, in contrast to S. cerevisiae x S. uvarum crosses, hybridization yield was positively affected by time of exposure to starvation, but not by staggered mating. In laboratory-scale fermentation trials at 20°C, one triple S. cerevisiae x S. eubayanus x S. uvarum hybrid showed a heterotic phenotype compared with the parents. In 2L wort fermentation trials at 12°C, this hybrid inherited the ability to consume efficiently maltotriose from NBRC1948 and, like the sourdough S. cerevisiae parent, produced appreciable levels of the positive aroma compounds 3-methylbutyl acetate (banana/pear), ethyl acetate (general fruit aroma) and ethyl hexanoate (green apple, aniseed, and cherry aroma). Based on these evidences, the phenotype-centered approach appears promising for design of de novo lager beer hybrids and may help to diversify aroma profiles in lager beers.

In Brazil, sucrose-rich broths (cane juice and/or molasses) are used to produce billions of liters of both fuel ethanol and cachaça per year using selected Saccharomyces cerevisiae industrial strains. Aiming to improve this disaccharide fermentation, we have first determined the genetic characteristics, by array comparative genomic hybridization, of a group of 8 fuel ethanol and 5 cachaça industrial yeasts that tend to dominate the fermentors during the production season. The widespread presence of SUC genes encoding invertase at multiple telomeres has been shown to be a common feature of both baker’s and distillers’ yeast strains and is postulated to be an adaptation to sucrose-rich broths. Our results show that only two strains (one fuel ethanol and one cachaça yeast) have amplification of the SUC genes, with high invertase activity. The other industrial yeast strains had a single SUC locus (SUC2) in their genome, with different patterns of invertase activity. These results indicate that invertase activity probably does not limit sucrose fermentation during fuel ethanol and cachaça production by these industrial strains. Using this knowledge, we changed the mode of sucrose metabolism of an industrial strain by avoiding extracellular invertase activity, overexpressing the intracellular invertase, and increasing its transport through the AGT1 permease. This approach allowed higher ethanol production from sucrose by the modified industrial yeast when compared to its parental strain.

Red fruits such as grapes and blueberries are known for their high content of bioactive compounds and antioxidants. In Mediterranean winemaking, traditional sun drying can be replaced by controlled airflow chamber drying which provides better quality, higher phenolic content, and increased antioxidants. This study aimed to increase the sugar content and phenolic compounds of the must by drying the fruits to 50% of their original moisture content. Two musts were prepared by combining fresh grapes and dried blueberries (M1), and the other using dried grapes and fresh blueberries (M2), followed by fermentation at 25 ºC with M05 Mead and X5 yeast strains. Must M2 showed the highest level of phenolic compounds, A520, total anthocyanins and antioxidant activity. During fermentation, the anthocyanin content increased mainly in the dried blueberry macerates, which raised between 4 to 5.5-fold. More bioactive compounds were extracted from wines produced by yeast inoculation despite shorter maceration times. Sensory analysis demonstrated consumer acceptance of the wines in terms of color, flavor and aroma. In conclusion, the use of red grapes in the production of blueberry red wine proved to be effective, providing sugars and higher must yields, while the dried fruits improved the fermentable sugar content and the levels of bioactive compounds, increasing the antioxidant capacity of the resulting red fruit wines.

A Brewer Spent Grain (BSG), the most abundant by-product of the brewing process, is mainly used in animal feed and biogas production. The aim of the present work is BSG valorization by yeast fermentation to produce carotenoids. R. mucilaginosa CBS316 was cultured on BSG syrups, supplemented or not with YE and PEP (s-Fsa and Fsa). The biomass and carotenoid production were optimized in flasks and then scaled-up in bioreactor. In flask tests, the maximum biomass concentration was 5.3 and 6.2 g/L for Fsa and s-Fsa cultures, respectively. The maximum carotenoid production was 833 μg/L in Fsa and 1159 μg/L in s-FSA. Considering that the highest specific productivity value (P 7.16 μg/g*h) was obtained with Fsa syrup, this medium was selected for the process scale-up. In bioreactor tests, the maximum concentrations of biomass (4.29 g/L) and carotenoid (396 ug/L) was reached at 72 hours of fer-mentation, 24 h later than in flask. The total carotenoid produced was 524 and 150 ug in bioreactor and flask cultures, respectively. The carotenoids production by R. mucilaginosa was obtained both in Fsa and s-Fsa syrups. The results confirm the possibility to use the syrup as it is, Fsa, thus limiting the process costs.

The aim of this experiment was to investigate the effects of moisture content, additives and their interactions on the fermentation quality, aerobic stability and in vitro digestibility of mixed silage of amaranth and corn meal. Amaranths and corn meal were mixed in mass ratios of 69:31, 76:24 and 84:16, and the silage water content was adjusted to 60% (W1), 65% (W2) and 70% (W3), respectively. Silage treatments included no additives (U), the addition of Lactobacillus plantarum (L), the addition of cellulase (E), and the addition of Lactobacillus plantarum + cellulase (M) mixed reagents. The results showed that pH and ammonia nitrogen (NH3-N/TN) levels were significantly lower (p < 0.05) in W1 compared to W2 and W3. At the same time, dry matter (DM), organic matter (OM), in vitro dry matter digestibility (ivDMD), in vitro dry matter digestibility (ivDMD), in vitro organic matter digestibility (ivOMD), and in vitro crude protein digestibility (ivCPD) were significantly increased (p < 0.05). Meanwhile, the aerobic stability of mixed silage of seed amaranth and corn meal decreased with increasing water content. The aerobic stability of the L, E, and M treatment groups was improved by 15, 105, and 111 hours, respectively, compared to the control group at W1. pH and NH3-N/TN levels were lower in the addition of E (E and M) than in the absence of E (U and L) (3.73, 20.1 g kg−1DM vs. 3.64, 22.9 g kg−1 DM, p < 0.05).NDF and ADF were significantly lower with the addition of E than without the addition of E (598 g kg−1 DM, 145 g kg−1 DM vs. 632 g kg−1 DM, 160 g kg−1 DM, p < 0.05). However, CP, ivDMD, ivOMD, and ivCPD were significantly higher (p < 0.05). AA and NH3-N/TN were significantly reduced (p < 0.05) with the addition of L (L and M) compared to without the addition of (U and E). In conclusion, the best fermentation quality, in vitro digestibility and aerobic stability of amaranth and cornmeal mixed silage treated with Lactobacillus plantarum + cellulase (M) were achieved at 60% water content.

Europium is one of the most reactive lanthanides and humans use it in many different applica-tions, but we still know little about its potential toxicity and cellular response to its exposure. Two strains of the eukaryotic microorganism model Tetrahymena thermophila adapted to high concentrations of two Eu(III) compounds (EuCl3 or Eu2O3) were obtained and compared to a control strain and cultures treated with both compounds. In this ciliate, EuCl3 is more toxic than Eu2O3. LC50 values show that this microorganism is more resistant to these Eu(III) compounds than other microorganisms. Oxidative stress originated mainly by Eu2O3 is minimized by overexpression of genes encoding important antioxidant enzymes. The overexpression of metallothionein genes under treatment with Eu(III) compounds supports the possibility that this lanthanide may interact with the -SH groups of the cysteine residues from metallothi-oneins and/or displace essential cations of these proteins during their homeostasis function. Both lipid metabolism (lipid droplets that fuse with europium-containing vacuoles) and au-tophagy are involved in the cellular response to europium stress. Bioaccumulation, together with a possible biomineralization to europium phosphate, seems to be the main mechanism of Eu(III) detoxification in these cells.

The wellness economy has developed rapidly recently, particularly in wellness tourism. Wellness tourism transforms sustainable strategy through the availability of extended multi-actor engagement in service innovation to further the tourism experience. Therefore, when operating a wellness tourism destination, it is necessary to encourage multi-actor engagement in service innovation to achieve sustainable development. This paper introduces a mixed research method with the SWOT-AHP-QSPM, as it qualitatively analyzes the advantages, disadvantages, opportunities, and threats, constructs an evaluation index system, conducts quantitative analysis using the analytic hierarchy process, determines the index weight and weighted score, and calculate the sustainable development strategy, which is appropriate for wellness development of Chongqing Tea Mountain and Bamboo Forest National Forest Park. Purposive field surveys and personal interviews were conducted in Chongqing forest tourism destinations, involving 356 effective respondents. The research reveals that multi-actor engagement significantly enhances sustainable development. These findings bridge service innovation knowledge gaps and provide valuable insights for forest tourism destination management and sustainable policy strategies to elevate multi-actor engagement in forest sites.

Objective: The aim of this study was to optimize the fermentation process of Radix Ranunculi ternate via microbial fermentation and analyze the changes in the contents of the main components, the antioxidant and hypoglycemic capacities of the extract before and after fermentation. Methods: The solid-state fermentation process was optimized using single-factor tests and the response surface method, with the yield of the alcohol extract of R. ternate as an evaluation index. Results: The best fermentation process was optimized using solid-state endophytic fungus fermentation technology as follows: strain addition ratio of Chaetomium globosum/Fusarium equiseti =1:1, fermentation for 5 d, sieve size of 40 mesh, liquid/material ratio of 0.8:1 mL·g−1, fermentation temperature of 31 °C, and inoculation amount of 7.5%. Under the optimized conditions, the contents of the water-soluble extract and total polysaccharides decreased by 12.71% and 12.95%, respectively, in the fermentation, The contents of ethanol-soluble-extract, flavonoids, saponins, polyphenols, organic acids, and total amino acids of fermented R. ternate increased by 19.77%, 57.14%, 79.67%, 14.29%, 17.63% and 3.82%, respectively, and the scavenging rate for DPPH, ABTS+, ·OH free radicals and inhibitory rate for α-amylase also increased by 19.02%, 14.17%, 7.53% and 34.54%, respectively, compared with unfermented R. ternate. Conclusions: Solid-state fermentation opens new avenues for the development and application of R. ternate as natural antioxidant and hypoglycemic food.

Partially state-owned firms are common in both developed and developing countries and contribute around 10% of the world's GDP. This has led to a rich literature exploring various aspects of competition between such “mixed”-ownership firms. This paper contributes to the existing literature by studying the effects of mixed ownership on the intensity of competition in oligopoly markets. Our findings indicate that state ownership exerts an anti-competitive influence, with a higher state ownership share resulting in less aggressive behavior by firms in the product market. This result implies that industries with a higher proportion of state ownership yield lower total surplus. Further, when considering factors such as taxes, soft budget constraints, and weak property rights, the anti-competitive effects of state ownership are accentuated.

This experiment aimed to investigate the impact of malic acid (MA) and citric acid (CA) on the nutritional composition, fermentation quality, rumen degradation rate, and microbial diversity of a mixture of apple pomace and corn protein powder during ensiling. The experiment used apple pomace and corn protein powder as raw materials, with four groups: control group (CON), malic acid treatment group (MA, 10g/kg), citric acid treatment group (CA, 10g/kg), and citric acid + malic acid treatment group (MA, 10g/kg + CA, 10g/kg), subjected to mixed ensiling for 60 days. The results indicated: (1) Compared to the CON group, the crude protein content significantly increased in the MA, CA, and MA+CA groups (p < 0.05), with the highest content observed in the MA+CA group. The addition of MA and CA effectively reduced the water-soluble carbohydrate (WSC) content (p < 0.05). Simultaneously, the CA group showed a decreasing trend in NDFom and hemicellulose content (p = 0.08; p = 0.09). (2) Compared to the CON group, the pH significantly decreased in the MA, CA, and MA+CA groups (p < 0.01), and the three treatment groups exhibited a significant increase in lactic acid and acetic acid content (p < 0.01). The quantity of lactic acid bacteria increased significantly (p < 0.01), with the MA+CA group showing a more significant increase than the MA and CA groups (p < 0.05). (3) Compared to the CON group, the in situ dry matter disappearance (ISDMD) significantly increased in the MA, CA, and MA+CA groups (p < 0.05). All three treatment groups showed highly significant differences in in situ crude protein disappearance (ISCPD) compared to the CON group (p < 0.01). (4) Good's Coverage for all experimental groups was greater than 0.99, meeting the conditions for subsequent sequencing. Compared to the CON group, the Shannon index significantly increased in the CA group (P < 0.01), and the Simpson index increased significantly in the MA group (p < 0.05). However, there was no significant difference in the Chao index among the three treatment groups and the CON group (p > 0.05). At the genus level, the abundance of Lentilactobacillus in the MA, CA, and MA+CA groups was significantly higher than in the control group (p < 0.05). PICRUSt prediction results indicated that the metabolic functional microbial groups in the CA and MA treatment groups were significantly higher than in the CON group (p < 0.05), suggesting that the addition of MA or CA could reduce the loss of nutritional components such as protein and carbohydrates in mixed ensilage. In conclusion, the addition of malic acid and citric acid to a mixture of apple pomace and corn protein powder during ensiling reduces nutritional losses, improves fermentation quality and rumen degradation rate, enhances the diversity of the microbial community in ensiled feed, and improves microbial structure. The combined addition of malic acid and citric acid demonstrates a superior effect.

Heavy metal accumulation in soils used for agricultural purposes is a major issue as it can affect the growth of plants and contain health risks as plants containing heavy metals are taken as food. This study provided the potential of three Bacillus pumilus strains (C-2PMW-8, C-1 SSK-8, and C-1PWK-7) for mitigation of the health factors related to heavy metals on lettuce (Lactuca sativa) grown in chromium contaminated soil from activities related to mining. Lettuce seeds inoculated with B. pumilus strains were grown in pots with garden soil amended with varying amount of chromite mine soil (2.27%, 4.65%, 7.14%). Growth indicators like days to germination, percentage of germination, leaf weight, leaf area and rate of survival decreased with incremental soil chromium levels but improved highly with inoculation of B. pumilus compared to uninoculated controls. When B. pumilus strains were provided, there was also a decreased level in the buildup of chromium in soil as well as lettuce shoots. The results explains that plant growth-promoting B. pumilus strains may be able to lower down the detrimental effects of chromium on lettuce development, hence increment in the effectiveness of phytoremediation.

To address the insufficient mechanical properties of calcined phosphogypsum, this research aims to investigate the utilization of basalt fiber and calcium carbonate whiskers as reinforcing agents. The study examines their impact on the flexural and compressive strength, toughness, water resistance, and tensile strength of calcined phosphogypsum. In the individual tests, basalt fibers with different lengths (3 mm, 6 mm, 9 mm, and 18 mm) were added at dosages of 0%, 0.5%, 1.0%, and 1.5% respectively. The findings demonstrate that basalt fiber effectively enhances the flexural and compressive strength, toughness, and tensile strength of calcined phosphogypsum, albeit potentially compromising water resistance. Among the various fiber lengths, the 6 mm fibers exhibit the most advantageous influence on the performance of calcined phosphogypsum. A subsequent test was conducted to investigate the impact of cross-scale fibers on the properties of calcined phosphogypsum by mixing 6 mm basalt fibers and calcium carbonate whiskers. The results illustrate that calcium carbonate whisker refines the pores, thereby improving the flexural strength and toughness of calcined phosphogypsum. Furthermore, it compensates for the water resistance limitations associated with the sole utilization of basalt fiber while further augmenting the tensile strength and strain capacity. However, it should be noted that increasing the dosage of both calcium carbonate whiskers and basalt fibers concurrently leads to increased porosity of phosphogypsum and reduced compressive strength.

Fe-(9, 19, 28, 37)wt.% Cr alloys were corroded at 700 and 800 ^oC for 70 h under 1 atm of N₂, 1 atm of N₂/3.2%H₂O-mixed gas, and 1 atm of N₂/3.1%H₂O/2.42%H₂S-mixed gas. The corrosion rate of Fe-9Cr alloy increased with the addition of H₂O, and furthermore with the addition of H₂S in N₂ gas. Fe-9Cr alloy was always nonprotective. In contrast, Fe-(19, 28, 37) wt.%Cr alloys were protective in N₂ and N₂/H₂O-mixed gas because of formation of the Cr₂O₃ layer. They were, however, nonprotective in N₂/H₂O/H₂S-mixed gas because sulfidation dominated to form the outer FeS layer and the inner Cr₂S₃ layer containing some FeCr₂S₄.

Soot combustion experiments with 5%O2/He were conducted using model soot, and four distinct compositions of CexPr1-xO2-δ oxides were prepared, varying nominal cerium com-positions (x=0, 0.2, 0.3, and 1). The catalyst samples were comprehensively characterized using techniques such as XRD, Raman spectroscopy, HR-TEM, N2 adsorption at −196 °C, XPS, O2-TPD, H2-TPR, and work function measurements. The Pr-rich compositions, rang-ing from Ce0.3Pr0.7O2-δ to PrO2-δ, resulted in a significant increase in the total evolved O2 amounts and enhanced catalyst reducibility. However, a decrease in textural properties of the catalysts was noted, being particularly important for the pure praseodymia under the synthesis route conducted. The catalytic activity was investigated under two contact modes of mixing between soot and catalyst: loose and tight. The results revealed that the catalytic performance is associated with the surface contact in tight contact mode and with the combination of surface/subsurface/bulk oxygen mobility and the BET surface ar-ea in loose contact mode. Notably, the T10 and T50 parameters were achieved at much lower temperatures than the uncatalyzed soot combustion, even under loose contact conditions. Specifically, the 50% conversion was achieved at 511℃ and 538℃ for Ce0.3Pr0.7O2 and Ce0.2Pr0.8O2, respectively. While no direct correlation between catalytic activity and work function was observed a significant relationship emerges between work function values and the formation of oxy-gen vacancies, whatever the conditions used for these measurements. On the other hand, the ability to generate a high population of oxygen vacancies at low temperatures, rather than the direct activation of gas-phase O2, influences the catalytic performance of Pr-doped ceria catalysts, highlighting the importance of surface/subsurface oxygen vacancy generation, which was the parameter which showed better correlation with the catalytic activity, whatever the soot conversion value or the mode of contact considered.

The fermentation of syngas is an attractive technology that can be integrated with gasification of lignocellulosic biomass. The coupling of these two technologies allows for treating a great variety of raw materials. Lignin usually hinders microbial fermentations; thus, the thermal decomposition of the whole material into small molecules allows for the production of fuels and other types of molecules using syngas as substrate, a process performed at mild conditions. Syngas contains mainly hydrogen, carbon monoxide and carbon dioxide in varying proportions. These gases have a low volumetric energy density, resulting more interesting its conversion into higher energy density molecules. Syngas can be transformed by microorganisms, thus avoiding the use of expensive catalysts, which may be subject to poisoning. However, the fermentation is not free of suffering from inhibitory problems. The presence of trace components in syngas may cause a decrease in fermentation yields or cause a complete cessation of bacteria growth. The presence of tar and hydrogen cyanide are just examples of this fermentation's challenges. Syngas cleaning impairs significant restrictions in technology deployment. The technology may seem promising, but it is still far from large-scale application due to several aspects that still need to find a practical solution.

Cercospora beticola Sacc. is an ascomycete pathogen that causes Cercospora leaf spot in sugar beets (Beta vulgaris L.) and other related crops. The disease symptoms are characterized by circular lesions with an ash-gray centre and dark brown to purple margins on the leaves. It can lead to significant yield losses if not effectively managed. This study aims to characterize and assess the impact of bacterial biological control agents against C. beticola. Following a dual-culture screening, 18 bacteria exhibiting over 50% inhibition were selected, with six of them demonstrating more than 80% control. Identification of the bacteria was carried out through sequencing of the16S rDNA, revealing potentially 12 species belonging to six genera including Bacillus which was represented by four species. Additionally, the bacteria underwent in-depth characterization of biochemical and molecular properties, as well as plant-growth promotion. PCR detection of genes responsible for producing antifungal metabolites revealed that 83%, 78%, 89%, and 56% of the selected bacteria possessed the bacillomycin, iturin, fengycin, and surfactin encoding genes, respectively. Infrared spectroscopy analysis confirmed the presence of lipopeptide structure in the bacteria supernatant filtrate. Subsequently, the bacteria were assessed for their effect on sugar beet plants in controlled conditions. The bacteria exhibited notable capabilities, promoting growth in both roots and shoots, resulting in significant increases in root length, weight, and shoot length. The field experiment with four bacterial candidates demonstrated good performance against C. beticola compared to difenoconazole fungicide. These bacteria played a significant role in disease control, achieving a maximum efficacy of 77.42%, slightly below the 88.51% efficacy attained by difenoconazole. Additional field trials are necessary to verify the protective and growth-promoting effects of these candidates, whether applied individually, combined in consortia, or integrated with chemical inputs in sugar beet crop production.

This study evaluated batch fermentation modes, namely, separate hydrolysis and fermentation (SHF), Quasi-simultaneous saccharification and fermentation (Q-SSF), and simultaneous saccharification and fermentation (SSF), and fermentation conditions, i.e., enzyme and yeast loadings, nutrient supplementation and sterilization, on high titer bioethanol production from SPORL-pretreated Douglas-fir forest residue without detoxification. The result indicated Q-SSF and SSF were obviously superior to SHF operation in terms of ethanol yield. The enzyme loading showed a strong positive correlation between enzyme loading and the ethanol yield. The nutrient supplementation and sterility was not necessary for ethanol production from SPORL-pretreated Douglas-fir. The yeast loading showed no significant influence on the ethanol yield for typical SSF conditions. The terminal ethanol titer of 43.2 g/L, or 75.1% theoretical based on glucose, mannose, and xylose theoretical was achieved when SSF was conducted at the condition of following: whole slurry solids loading of 15%, enzyme loading of 20 FPU/g glucan, 1.8 g/kg (wet) yeast loading, without nutrition supplementation and sterilization, at 38°C, on shake flask at 150 rpm for 96h. It is believed that with mechanical mixing, enzyme loading can be substantially reduced with affect ethanol yield by using a long fermentation time.

The numerical simulation of entropy generation at mixed natural free and forced convection flow in a square lid-driven cavity filled with an eletrically conducting binary fluids saturated porous media in the presence of a magnetic field is performed in this investigation. Both the top and bottom horizontal walls of the cavity are kept at constant and different temperatures while the left and right vertical walls are adiabatic and insulated. The magnetic field is applied in normal direction to the cavity. The Darcy model, including Brinkman term relative to viscous effects and Forchheimer term due to inertial forces, is used for the momentum equations, and the SIMPLER algorithm, based on the control volume finite-element method approach is used to solve the pressure –velocity coupling. The flow pattern and the heat transfer characteristics inside the cavity are presented in the form of isotherms, streamlines and isentropic lines and average Nusselt number trend. The entropy generation rate is determined for various values of Darcy number (10^-3 ≤ Da ≤ 1) and for a range of Hartmann number (0 ≤ Ha ≤ 10²). This study is limited to Pr = 7 which corresponds to water. It is observed that entropy generation is very affected by the variations of dimensionless control parameters considered.

Background: Health insurance (HI) run by government is providing health care service to large population. Due to poor accountability, participation and sustainability, cooperative health insurance is becoming more popular and effective in low and middle income and some high-income countries too. In Nepal, there are public and cooperative HI is in practice. The aim of this study is to compare the effectiveness of public (government) and cooperative HI in relation to benefit packages, population coverage, inclusiveness, health care utilization, and promptness for treatment in these two health insurance models in Nepal. Method: This is an institution based concurrent mixed study consists of qualitative and quantitative variables from public and cooperative groups. We included all public HI operated by government hospitals and cooperatives groups those purchased hospital service in contract. Two separate study tools were applied to access the effectiveness of insurance models. The key questions were asked for the representatives of government and private health insurance. The numeric information consisted of in quantitative data and subjective response was included in qualitative approach. Descriptive statistics and Mean Whitney U test was applied in numeric data and qualitative information were analyzed by inductive approach Results: The study revealed that new enrolment was not increased, health care utilization rate was increased and the benefit package was almost same in both groups. The overall inclusiveness was higher for the government HI, but enrolment from the religious minority, proportion of negotiated amount during treatment were significantly higher (p<0.05). During illness, the response time to reach hospital was significantly faster in cooperative health insurance than government health insurance. Qualitative findings showed that level of participation, accountability, transparency and recording system was better in cooperative health insurance than public. Conclusion: Cooperative HI could be more sustainable and accountable to the community for all; low, middle and high-income countries.

Mycobacterium tuberculosis was responsible for 1.3 million deaths worldwide in 2022. Bacillus Calmette-Guérin (BCG) is the only licensed vaccine against tuberculosis (TB); however, it has limited protective efficacy in adults. In this study, we constructed a recombinant vaccinia virus expressing Ag85B from M. tuberculosis using a novel attenuated vaccinia virus (KVAC103). We then analyzed the immunogenicity of prime-boost inoculation strategies using recombinant KVAC103 expressing Ag85B (rKVAC85B) compared to BCG. In both the rKVAC85B prime-boost and the BCG prime-rKVAC85B boost inoculation regimens, rKVAC85B induced the generation of specific IgG and the secretion of IFN-γ by immune cells. In vitro analysis of Mycobacterium growth inhibition revealed a comparable immune-mediated pattern of outcomes. Furthermore, bacterial loads in the lungs were significantly lower in mice inoculated with the BCG prime-rKVAC85B boost, than in the BCG-only group following a rechallenge infection with both H37Rv and HN878 strains of M. tuberculosis. These findings collectively suggest that KVAC103, incorporated into a viral vector, is a promising candidate for a novel TB vaccine platform that is effective against multiple TB strains, including H37Rv and HN878, and that rKVAC85B effectively stimulates immune responses against TB infection.

The lactic acid bacteria Streptococcus thermophilus and Lactobacillus helveticus are commonly used as starter cultures in dairy product production. This study aimed to investigate the characteristics of fermented milk using different ratios of these strains and analyze the changes in volatile compounds during fermentation and storage. The 10:1 ratio of Streptococcus thermophilus CICC 6063 to Lactobacillus helveticus CICC 6064 showed optimal fermentation time (4.2 h), viable cell count (9.64 log10 colony-forming units/mL), and sensory evaluation score (79.1 points). In total, 56 volatile compounds were identified and quantified by solid-phase microextraction and gas chromatography–mass spectrometry (SPME-GC-MS), including aldehydes, ketones, acids, alcohols, esters, and others. Among these, acetaldehyde, 2,3-butanedione, 2,3-pentanedione, 2-heptanone, acetoin, and 1-heptanol had the highest odor activity values and contributed to the characteristic flavor of fermented milk. The concentrations and types of key flavor compounds reached their peak at the end of fermentation. Throughout the fermentation and storage process, the levels of 2,3-pentanedione and acetoin exhibited synergistic dynamics. These findings enhance our understanding of the chemical and molecular characteristics of milk fermented with Streptococcus thermophilus and Lactobacillus helveticus, providing a basis for improving the flavor and odor of dairy products during fermentation and storage.

Marine yeasts have versatile applications in industrial, medical and environmental fields, but have received little attention compared to terrestrial yeasts and filamentous fungi. In this study, a phylogenetic analysis of 11 marine-derived yeasts was conducted using internal transcribed spacers and nuclear large subunit rDNA, and their bioactivities, such as antioxidant, antibacterial, and tyrosinase inhibition activities, were investigated. The 11 marine-derived yeasts were identified to belong to seven species including Geotrichum candidum, Metschnikowia bicuspidata, Papiliotrema fonsecae, Rhodotorula mucilaginosa, Vishniacozyma carnescens, Yamadazyma olivae, and Yarrowia lipolytica, and three strains of these were candidates for new species of the genera Aureobasidium, Rhodotorula, and Vishniacozyma. Most extracts showed antioxidant activity, whereas seven strains exhibited antibacterial activities against Bacillus subtilis. Only Aureobasidium sp. US-Sd3 among the 11 isolates showed tyrosinase inhibition. Metschnikowia bicuspidata BP-Up1 and Yamadazyma olivae K2-6 showed notable radical-scavenging activity, which has not been reported previously. Among the isolates, Aureobasidium sp. US-Sd3 exhibited the highest antibacterial and tyrosinase inhibitory activities. Overall, our results demonstrate the potential of marine-derived yeasts as a source of bioactive compounds for improving industrial applications.

Society considers wine as a special product among food and beverages because of its high gastronomical value and its positively distinctive quality. In recent years, philosophies of the agricultural techniques and development of the oenological technology have been focused on the reduction of wasteful, "polluting" elements, and trends are moving towards an environmental friendly approach. Due to the stricter regulations and rules (with the limited amount and selection of the permitted chemicals) resistant, also known as interspecific or innovative grape varieties can be the ideal basic materials of alternative cultivation technologies. In terms of variety selection, innovative varieties can be equivalent to international varieties, although organically their quality could not compete with them. These grapes are more resistant to various fungal diseases and infections than international varieties. Well-founded analytical and organoleptic results have to provide the scientific background of resistant varieties, as these cultivars with the environmental friendly cultivation techniques, could be the raw material of the future.

Kratom (Mitragyna speciosa) is a Southeast Asian plant containing various alkaloids which induce pharmacological effects in humans. In Western countries, online vendors sell a variety of different kratom strains which are marketed to have distinct effect profiles. However, as of yet such marketing claims are unsubstantiated, and therefore the current study investigated whether differently colored kratom products can induce unique effects, as self-reported by users. 644 current kratom users were anonymously surveyed to compare the self-reported effects and motivations for using kratom products sold as red, green, and white strains. Most of the survey respondents were customers of the same kratom vendor, the products of which had been assessed for their alkaloid content by an independent laboratory. The survey respondents reported distinct subjective experiences for different kratom strains, in a manner congruent with common marketing descriptions. However, the product analyses revealed no significant cross-strain differences in alkaloid content, suggesting that the reported effect differences might be disproportionally influenced by marketing narratives and anecdotal reports. Future studies should engage a more diverse population and include kratom strains from various vendors. Controlled, blinded experiments could assess whether the reported effect differences stem from a placebo effect or from alternative factors, e.g. minor alkaloids and terpenes.

Microorganisms belonging to the Lactobacillus genus are naturally associated or deliberately added to fermented food products and are widely used in probiotic food additives and supplements. Moreover these bacteria normally colonize mouth, gastrointestinal (GI) tract and female genitourinary tract of humans. They exert multiple beneficial effects and are regarded as safe microorganisms. However, infections caused by lactobacilli, mainly endocarditis, bacteremia and pleuropneumonia occasionally occur. The relevance of Lactobacillus spp. as opportunistic pathogens in humans and related risk factors and predisposing conditions are illustrated in this review article with more emphasis on the species L. rhamnosus, that has been more often involved in infection cases. The methods used to identify this species in clinical samples, to distinguish strains and to evaluate traits that can be associated to pathogenicity, as well as future perspectives for improving the identification of potentially pathogenic strains are outlined.

Microorganisms present in fermented foods and beverages play a vital role in enhancing both nutritive and non-nutritive components. During the fermentation, microorganisms can either naturally occur in the food through spontaneous fermentation or be intentionally added as starter cultures with a variety of species and strains. In this review, the focus is placed on micronutrients, such as vitamins and minerals, and three major macromolecules – prebiotics, bioactive peptides, and functional fatty acids. During fermentation, non-digestible carbohydrates like fructooligosaccharides and galactooligosaccharides are synthesized, imparting prebiotic properties that support gut health. Enzymatic hydrolysis of proteins by microorganisms yields bioactive peptides with diverse health benefits. Furthermore, fermentation facilitates the formation of fatty acids, including short-chain fatty acids and conjugated linoleic acid, potentially exerting physiological effects. By harnessing the power of microbial fermentation, nutrient-rich and bioactive compounds can be enhanced, offering a promising approach to address global malnutrition concerns.

This study provides a comprehensive investigation of lactic acid bacteria (LAB) isolated from Argentinean Capsicum annum L. The research covers important aspects, including genotypic characterization, bacterial stress tolerance, adhesion ability, safety evaluation, and functional and technological properties. The predominant isolates were identified as Lactilactobacillus curvatus and Lactiplantibacillus plantarum. Rep-PCR analysis grouped the isolates into 11 clonal groups. Lp. plantarum LVP 40 and LV 46; Levilactobacillus brevis LVP 41, Pediococcus pentosaseus LV P43, and Lt. curvatus LVP44, displayed both safety and resilience against adverse conditions such a slow pH, bile, and simulated gastric and intestinal juices. Moreover, the LAB strains exhibited high hydrophobicity and auto-aggregation percentages, NaCl tolerance, and substantial acidifying capacity. LAB supernatants demonstrated promising surfactant and emulsifying properties. Likewise, they differentially inhibited Staphylococcus aureus and Pseudomonas aeruginosa biofilms, showcasing their potential as antipathogenic agents. Noteworthy some strains displayed considerable co-aggregation with these pathogens, and several isolates showed an effective antimutagenic and detoxifying power, further emphasizing their multifaceted capabilities. Five pepper bacterial strains showcased probiotic properties, suggesting their potential for gut health enhancement. In summary, these LAB strains hold promise as vegetable fermentation starters, contributing to food safety and versatile applications in food science.

Arthrospira platensis, commercially known as Spirulina, is a fresh-water cyanobacterium that is gaining even more attention in the last years due to its high biological and nutritional value. For this reason, it has been employed in several food applications, to obtain or enhance functional and technological properties of cheese, yogurt, bread, cookies or pasta. The aim of this work was to evaluate the potential boosting effect of two different concentrations (0.25% and 0.50%) of Arthrospira platensis on the fermentation capability of several starter LAB strains, 1 probiotic and 4 commercial mix culture. These strains were used to ferment three different substrates and their fermentation behaviors were evaluated by impedance analyses together with rheological and color measurements. It was demonstrated that the booster effect took place, but it was variable and dependent not only on the strain or mix culture used, but also on the substrate and Arthrospira platensis concentration. Also, rheological and color modifications were found to be dependent of these factors.

The paper presents an extensometer designed to measure two mechanical strains at the same time—one from tensile load and the other from torsion load. Strain transducers provide different electric signals, which, after calibration, lead to the simultaneous measurement of linear (ε) and angular (γ) strains. Each of these two signals depends on the measured process and is not influenced by the other strain process. This extensometer is designed to be easily mounted on the sample with only two mounting points and can be used to measure the combined cyclical fatigue of tensile and torsional loadings. This extensometer has two bars—one rigid, reported at the resulting stress points, and one elastic and deformable. The elastic deformable bar has two beams with different orientations. When the sample is deformed, both beams are loaded by two bending moments (perpendicular to each other and both perpendicular on the longitudinal axis of the bars).

Fermented foods play an important role in the human diet and particularly so in under-resourced environments where cold preservation is not attainable due to irregular supply of electricity. Fermented foods are reported to support gut health by contributing probiotics and the purpose of this study was to investigate the microbial diversity and metabolic potential of spontaneous millet fermentation. The literature in the field was reviewed and analysis conducted on publicly available Sequence Read Archive (SRA) datasets. Quality analysis was performed with FastQC, and operational taxonomic units (OTUs) generated using Quantitative Insights Into Microbial Ecology (QIIME2) and Divisive Amplicon Denoising Algorithm (DADA2) pipeline with Greengenes as the reference database. Metagenomics and pathways analysis were performed with Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt2). Statistical analysis and visualization were accomplished with Statistical Analysis of Metagenomic Profiles (STAMP). At the family taxonomic level, there were significant differences in the relative abundances of bacteria involved in the spontaneous fermentation of millet namely, Lactobacillaceae, Leuconostocaceae, and Clostridiaceae in one dataset. The incidence of Lactobacillaceae and Bifidobacteriaceae suggest the probiotic characteristic of fermented millet. The datasets were collected with fermentations mediated by autochthonous microorganisms and the presence of some potential pathogens such as Enterobacteriaceae, Clostridiaceae, Aeromonadaceae, Microbacteiaceae, Pseudomonadaceae, and Neisseriaceae suggest the need for standardization of fermentation approaches. The genomes show the potential to synthesize metabolites such as vitamins suggesting that the respective fermented foods can be further optimized to enhance nutritional benefits.

Paenibacilli are efficient producers of potent agents against bacterial and fungal pathogens, which are of great interest both for therapeutic applications in medicine as well as in agrobiotechnol-ogy. Lipopeptides produced by such organisms play a major role in their inactivation potential. In this work we investigated two lipopeptide complexes, the fusaricidins and the polymyxins, produced by Paenibacillus polymyxa strains DSM 32871 and M1 by MALDI-TOF mass spectrometry. The fusaricidins show potent antifungal activities and are distinguished by an unusual variabil-ity. For strain DSM 32871 we identified numerous yet unknown variants mass spectrometrically. DSM 32871 produces polymyxins of type E (colistins), while M1 forms polymyxins P. For both strains novel, but not yet completely characterized polymyxin species were detected, which pos-sibly are glycosylated. These compounds may be of interest therapeutically, because polymyxins attain increasing attention as last-resort antibiotics against multiresistant pathogenic Gram-negative bacteria. In addition, the volatilomes of DSM 32781 and M1 were investigated with a GC-MS approach using different cultivation media. Production of volatile organic com-pounds (VOCs) was strain and medium dependent. In particular, strain M1 manifested as an effi-cient VOC-producer that exhibited formation of 25 volatiles in total. A characteristic feature of Paenibacilli is the formation of volatile pyrazine derivatives.

Beer fermentation is typically monitored by periodic sampling and off-line analysis. In-line sensors would remove the need for time-consuming manual operation and provide real-time evaluation of the fermenting media. This work uses a low-cost ultrasonic sensor combined with machine learning to predict the alcohol concentration during beer fermentation. The highest accuracy model (R2=0.952, MAE=0.265, MSE=0.136) used a transmission-based ultrasonic sensing technique along with the measured temperature. However, the second most accurate model (R2=0.948, MAE=0.283, MSE=0.146) used a reflection-based technique without the temperature. Both the reflection-based technique and the omission of the temperature data are novel to this research and demonstrate the potential for a non-invasive sensor to monitor beer fermentation.

One route to reducing CO2 emissions is to improve the energy efficiency of machines. For example, conventional combustion engines are being downsized (and down-speeded), are now running on lower viscosity engine lubricants (such as SAE 0W-20 or lower viscosity grades) and often also have stop-start systems fitted (to prevent engine idling when the vehicle is stopped). Some of these changes result in higher levels of mixed and boundary friction, and so accurate estimation of mixed/boundary friction losses is becoming of increased importance, both for estimating friction losses, and also for estimating wear volumes. Traditional approaches to estimating mixed/boundary friction, which employ real area of contact modelling such as the Greenwood-Tripp model [1] are widely used, but recent experimental data suggests that these approaches underestimate mixed/boundary friction losses [2-5]. A new model has been developed, based on experimental data, which estimates the proportion of mixed/boundary lubrication, X, as a function of the value (where is the ratio of the oil film thickness separating the surfaces to the combined root mean square surface roughness of the interface surfaces). The precise equation that describes the way in which X varies with takes the form of a “reverse S-curve”, which makes physical sense since S-curves arise naturally in growth processes, and in mixed/boundary lubrication, the real area of contact of rough surfaces grows as the load (and 1/) increases. Numerical estimates of mixed/boundary lubrication losses in internal combustion engines are presented here and compared with published data [2,6]. In addition, the improved method described here is used to estimate both the financial cost of mixed/boundary lubrication for today’s passenger car fleet, and the CO2 emissions associated with these friction losses.

Nutrigenomics is the study of the interaction of nutrition and genes, focusing on the influence of nutrients on the genome, proteome, and metabolome, and how nutrition affects human health. In the context of nutrigenomics, bioactive components are dietary ingredients that may transmit information from the external environment and alter gene expression in the cell, and hence the overall function of the organism. It is critical to consider food not only as a source of energy and essential nutrients necessary for life and growth, but also as a factor impacting health/disease, biochemical processes, biochemical pathway activation and affecting the diversity of the gut microbiome. Antimicrobial resistance in pathogenic and commensal microorganisms has emerged as a major public health concern due to emerging antimicrobial resistance genes in E. coli isolates from pig, cattle, chicken, and turkey meat, against tetracycline, streptomycin, and sulfonamides. Also, Salmonella spp. and Campylobacter spp. have shown antibiotic resistance at farms and slaughterhouses, and in animal-based food products. A correlation has been proven between a critical nutrient-responsive signaling system and catabolite control of gene expression, and a two-component signaling system that drives antibiotic resistance in E. faecalis, revealing a previously unknown integration between the nutritional status of the cell and intrinsic antibiotic resistance. Moreover, different nutrigenomic approaches can be applied to mitigate possible emergence of antimicrobial resistance against novel antibiotics. However, little progress has been achieved in converting nutrigenomics information into clinical advice, so far.

Thesis Statement: The consumption of externally fermented foods acted as the initial metabolic trigger enabling hominid brain expansion. Because brain tissue is metabolically expensive, it is thought that the evolution of humans’ large brains was only possible through a concomitant reduction in the size of another expensive organ system, the gut. However, this gut reduction must have itself been made possible by dietary changes, the nature of which are still unclear. Here, we propose that the initial metabolic trigger of hominid brain expansion may have been the consumption of externally fermented foods. We define “external fermentation” as occurring outside the body, as opposed to the internal fermentation that occurs through the gut microbiome. This practice could have begun accidentally and with limited understanding, but over time, fermentation technologies may have become increasingly intentional, socially-transmitted, and culturally-reinforced. We detail the mechanisms by which external fermentation can mediate the evolution of increased brain size, as well as a reduction in gut size, by increasing the bioavailability of macro- and micronutrients while reducing digestive energy expenditure. Importantly, we calculate that the reduction in human gut size relative to modern apes is mainly due to a reduction in the colon, the site of internal fermentation. We also discuss the explanatory power of our hypothesis relative to others, including realistic plausibility in hominids with brains roughly the size of modern chimpanzees. Finally, we survey external fermentation practices across human cultures to demonstrate its viability across a huge range of environments, temperatures, and food sources. We close with suggestions for empirical tests.

Quantum entanglement is a fascinating topic with both theoretical and technological impacts. Yet, even in the simplest scenarios, there are still various open questions involving pure state and mixed state entanglement. Here we present the simplest entangled mixed states. We consider both qubits (quantum bits) and qudits (quantum digits). We first provide our simplest yet most important result for rank-2 bi-partite states, and we then generalize our results to go beyond rank-2 and to go beyond bi-partite states.

The most common form of mixed dementia (MixD) is constituted by abnormal protein deposits associated with Alzheimer´s disease (AD) that coexist with vascular disease. Although olfactory dysfunction is considered a clinical sign of AD-related de-mentias, little is known about the impact of this sensorial impairment in MixD at molecular level. To address this gap in knowledge, we have assessed olfactory bulb (OB) proteome-wide expression in MixD subjects (n=6) respect to neurologically intact controls (n=7). Around 9% of the quantified proteins were differentially expressed, pinpointing aberrant proteostasis involved in synaptic transmission, nucleoside monophosphate and carbohydrate metabolisms and neuron projection regeneration. In addition, net-work-driven proteomics revealed a modulation in cell-survival related pathways such as ERK, AKT and PDK1-PKC axis. Part of the differential OB protein set was not specific of MixD, being also deregulated across different tauopathies, synucleinopathies and tardopathies. However, the comparative functional analysis of OB proteome data between MixD and pure AD pathologies deciphered commonalities and differences between both related phenotypes. Finally, olfactory proteomics allowed to propose serum Prolow-density lipoprotein receptor-related protein 1 (LRP1) as a candidate marker to differentiate AD from MixD phe-notypes.

Strong-aroma Baijiu is a traditional Chinese spirit distilled from grains that undergo solid-state fermentation. At the end of each fermentation cycle, a significant quantity of Huangshui accumulates at the bottom of the fermentation pit, constituting one of the primary by-products in the brewing process of strong-flavor liquor. In this report, the chemical component analysis of Huangshui obtained from a strong-flavor liquor factory revealed a diverse array of organic acids, alcohols, esters, starch, reducing sugars, amino acids, and other compounds present in Huangshui. Additionally, two specialized solid-state fermentation models were developed to investigate the formation process of Huangshui. Our findings demonstrate a positive correlation between the yield of Huangshui and the initial moisture content of fermented grains. It is concluded that Huangshui is the percolate produced during the solid-state brewing process of strong-flavor liquor, with its water content originating from the initial moisture of fermented grains.

Calcareous sand deposits are widely distributed along the shoreline in ropical and subtropical regions. Microbially induced calcite precipitation treatment (MICP) is a relatively new method to improve the stiffness and strength of the soil. Little is known about the small-strain shear modulus and damping ratio of MICP-treated calcareous sand, which are two crucial parameters for the prediction of the dynamic behavior of soil. A series of resonant column tests are performed to investigate the dynamic performance of MICP-treated calcareous sand, with special attention paid to the influence of treatment duration and confining stress on the stiffness and damping characteristics. The relationship between the initial dynamic shear modulus and unconfined compressive strength is analyzed. Additionally, the empirical equations of the reference shear strain between treatment duration and confining stress are given. The G/G0 of MICP-cemented calcareous sand presents a higher strain sensitivity than that of untreated sand, and its attenuation pattern can be described by Hardin-Drnevich model. The σc has an apparent effect on the degradation characteristics of the dynamic shear modulus of MICP-treated calcareous sand with a low cementation level, however, its effect decreases with the increasing treatment duration. The relationship between the reference shear strain and the treatment duration and confining stress can be described by a power and a linear formula, respectively.

The global food production system faces several challenges, including significant environmental impacts due to traditional agricultural practices. The rising demands of consumers for food products that are safe, healthy, and have animal welfare standards have led to an increased interest in alternative proteins and the development of the cellular agriculture field. Within this innovative field, precision fermentation emerges as a promising technological solution to produce proteins with reduced ecological footprints. This review provides a summary of the environmental impacts related to the current global food production, and explore how precision fermentation can contribute to address these issues. Additionally, we will report on the main animal-derived proteins produced by precision fermentation, with a particular focus on those used in the food and nutraceutical industries. The general principles of precision fermentation will be explained, including strain and bioprocess optimization. Examples of efficient recombinant protein production by bacteria and yeasts, such as milk proteins, egg-white proteins, structural and flavoring proteins, will also be addressed, along with case examples of companies producing these recombinant proteins in a commercial scale. Through these examples, we will explore how precision fermentation supports sustainable food production and holds the potential for significant innovations in the sector.

Information on the nutritive value and in vitro fermentation characteristics of native shrubs in New Zealand is scant. This is despite their potential as alternatives to exotic trees and shrubs for sup-plementary fodder, and mitigation of greenhouse gas and soil erosion on hill country sheep and beef farms. The objectives of this study were to measure the in vitro fermentation gas production, predict parameters of in vitro fermentation kinetics and to estimate in vitro fermentation of volatile fatty acids (VFA), microbial biomass (MBM) and greenhouse gases of four native shrubs (Coprosma robusta, Griselinia littoralis, Hoheria populnea and Pittosporum crassifolium) and an exotic fodder tree species, Salix schwerinii. Total in vitro gas production was higher (p<0.05) for natives than S. schwerinii. Prediction using the single pool model resulted in biologically incorrect negative in vitro total gas production from the immediately soluble fraction of the native shrubs. However, the dual pool model better predicted in vitro total gas production and was in alignment with measured in vitro fermentation end products. In vitro VFA and greenhouse gas production from fermentation of leaf and stem material were higher (p<0.05), and MBM lower (p<0.05), for native shrubs com-pared to S. schwerinii. The lower in vitro total gas production, VFA and greenhouse gases produc-tion, and higher MBM of S. schwerinii may be explained by the presence of condensed tannins (CT), although this was not measured and requires further study. In conclusion, results from this study suggests that when consumed by ruminant livestock, the browsable native shrubs can provide adequate energy and microbial protein, and that greenhouse gas production from these species is within ranges reported for typical New Zealand pastures.

A large variety of fermentation products are used in food and feed production, but also in other industries, and many of these products are produced with genetically modified microorganisms (GMMs). In food and feed production, prominent examples are amino acids, vitamins, food and feed enzymes, colorants, non-caloric sweeteners and human milk oligosaccharides. From a regulatory perspective, fermentation products are typically produced under containment. This means that premises, equipment and work processes need to be designed to prevent or at least minimize release of GMMs into the environment. The fermentation products themselves should not contain any live cells of the GMM. Over the past years, there have been concerning developments, particularly in the European Union, stipulating that also absence of recombinant DNA might be interpreted as a regulatory requirement for fermentation products produced with GMMs. In this paper, we (i) attempt to place these developments into the historical context, (ii) sketch the potential negative repercussions for the food and feed industries, (iii) elaborate on the safety of recombinant DNA, and (iv) postulate that recombinant DNA should remain an integral part of the safety assessment of fermentation products but should not be misconstrued as a criterion for regulatory classification of products of biotechnology.

The emergence of antimicrobial resistance remains one of the greatest public health concerns. Biofilm formation has been postulated as a mechanism of microbial pathogens to resist antimicrobial agents. Lactic Acid Bacteria (LAB) and their metabolites have been proposed to combat bacterial biofilms, due to their antimicrobial activity. In this vein, the aim of the present study was to investigate the biofilm removal potential of cell-free supernatant (CFS) of five wild-type Lacticaseibacillus rhamnosus LAB strains, isolated from Greek natural products, in comparison to the commercial available L. rhamnosus GG strain, formed by common foodborne pathogens (Salmonella Enteritidis, Salmonella Typhimurium, Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus). Biofilm removal activity of LAB was assessed on two-days mature biofilm using a microtiter plate-based procedure. Both non-neutralized and neutralized CFSs removed biofilms in a concentration-dependent manner. Biofilm removal activity of the non-neutralized CFSs was significantly higher compared to the neutralized CFSs, as expected, ranging 40 – 90% and 10 – 78%, respectively. Biofilm removal efficiency of L. rhamnosus OLXAL-3 was signifanctly higher among the wild-type L. rhamnosus strains tested (20 – 100% v/v). In conclusion, our results suggested the great potential of application of wild-type Lacticaseibacillus rhamnosus strains CFSs as effective natural agents against pathogenic bacterial biofilms.

Excessive body fat is a major contributor to obesity, which is related to high blood, total cholesterol and triglycerides. Lemons (Citrus limon) are renowned for their antioxidant and lipid-lowering properties. First, Sugar-free fermentation demonstrated better results lower sugar content and was identified as the preferred fermentation method. Next, the effect of two doses of sugar-free fermented lemon peel juice (SF-FLPJ) on obesity was investigated. Sprague Dawley (male rats) were randomly divided into 4 groups and fed a standard feed or high-fat diet (HFD) to induce fatness for eight weeks. The two groups of rats fed an HFD were administered SF-FLPJ at 0.45 or 0.9 g/kg body weight daily for eight weeks. The obese rats administered 0.9 g/kg (body weight/day) of SF-FLPJ exhibited significant reductions in body fat mass and percentage; they also had lower blood cholesterol and triglyceride levels than the other groups. Additionally, the rats displayed diminished liver fat accumulation. The results suggest that SF-FLPJ can be a supplement in managing body fat, contributing positively to addressing obesity within the context of health management, and reduce obesity-associated issues.

Recently, the popularity of fermented foods has increased significantly. Lactofermented vegetable juice products are particularly important for our diet. The combination of the richness of the ingredients derived from vegetable juices and the products formed during lactic fermentation is a valuable source of nutrients. This study aimed to analyse the selected properties of fermented beetroot, carrot, and beetroot-carrot juices. The juices obtained from beetroot and carrot were fermented with bacterial strains Lactiplantibacillus plantarum or Levilactobacillus brevis. Density, extract, dry matter content, pH, total acidity, pigments (betalain or carotenoids), color, and LAB count were measured in the samples. The results obtained in this study showed that the used LAB strains were capable of fermenting the analyzed juices. It was proven that Lactiplantibacillus plantarum was a better strain for fermentation ion of vegetable juice. This might have been influenced by the fermentation temperature and the duration of the fermentation. However, this strain had a lower effect on carotenoids, which are water-insoluble pigments, than Lactiplantibacillus plantarum. The opposite observation was made for betalain, a water-soluble pigment with nitrogen in its structure.

Polyphenols and fermentable fibers have shown favorable effects on gut microbiota composition and metabolic function. However, few studies have investigated whether combining multiple fermentable fibers or polyphenols may have additive beneficial effects on gut microbial states. Here, an in vitro fer-mentation model, seeded with human stool combined from 30 healthy volunteers, was supplemented with blends of polyphenols (PP), dietary fibers (FB) or their combination (PPFB) to determine influence on gut bacteria growth dynamics and select metabolite changes. PP and FB blends independently lead to significant increases in the absolute abundance of select beneficial taxa, namely Ruminococcus bromii, Bifidobacterium spp., Lactobacillus spp. and Dorea spp. Total short chain fatty acid concentrations, relative to non-supplemented control (F), increased significantly with PPFB and FB supplementation, but not PP. Indole and ammonia concentrations decreased with FB and PPFB supplementation, but not PP alone while increased antioxidant capacity was only evident with both PP and PPFB. These findings demonstrated that, while the independent blends displayed selective positive impacts on gut states, combination of both blends provided an additive effect. The work outlines the potential of mixed substrate blends to elicit a broader positive influence on gut microbial composition and function to build resiliency toward dysbiosis.

Introduction. The use of probiotics demonstrate efficacy against obesity and metabolic syndrome (MetS). Detection effective probiotic strains for hyperglycemia and immunity associated with is important task. The aim of the study was to evaluate an influence of Lactobacillus casei IMV B-7280 separately and composition L. casei IMV B-7280 / Bifidobacterium animalis VKB / B. animalis VKL on the levels of blood glucose and immunity in obese mice. Meterials and methods. Obesity was induced by fat-enriched diet (FED) in male BALB/c mice. Obese mice were transferred to standard diet and received per os probiotic strains daily during 10 days. We measured tumor necrosis factor-a (TNF-a) in blood serum using enzyme-linked immunosorbent assay and functional activity of peritoneal exudate macrophages (PEMs). Glucose levels in blood was defined with glucometer. Results. We ascertained that all probiotic strains induced reducing mice weight and visceral fat, normalization of TNF-a production and functional activity of PEMs. Treatment with L. casei IMV B-7280 was associated with decreasing blood glucose levels. No normalization of glucose and TNF-a levels in obese mice, transferred to standard diet without probiotic treatment, although we revealed decreasing their weight and visceral fat and partial recover of functional activity of PEMs. Conclusions. Probiotic strain L. casei IMV B-7280 (separately) and composition L. casei IMV B-7280 / B. animalis VKB / B. animalis VKL can re-equilibrate metabolic and inflammation indices in mouse obesity model. L. casei IMV B-7280 alone was more efficient in decreasing glucose levels than composition of strains.

The aim of the paper is to investigate Nash equilibria and correlated equilibria of classical and quantum games in the context of their Pareto optimality. We study four games: the prisoner's dilemma, battle of the sexes and two versions of the game of chicken. The correlated equilibria usually improve Nash equilibria of games but require a trusted correlation device. We analyze the quantum extension of these games in the Eisert-Wilkens-Lewenstein formalism with the full SU(2) space of players’ strategy parameters. It has been shown that the Nash equilibria of these games in quantum mixed Pauli strategies are closer to Pareto optimal results than their classical counterparts. The relationship of mixed Pauli strategies equilibria and correlated equilibria is also analyzed.

This study investigates the impact of inoculating Torulaspora delbrueckii (Td) strains during the initial phase of Carbonic Maceration (CM) vinification, aiming to enhance the fermentative process and unique characteristics of CM wines. CM is a winemaking technique where whole bunches (without destemming and crushing) are enclosed in tanks filled with carbon dioxide, inducing intracellular fermentation. The study compares the effects of two Td strains on the MLF performance and sensory characteristics of CM wines, using both inoculated and spontaneous MLF strategies. Although general physicochemical parameters remained consistent across conditions, organoleptic attributes showed significant differences due to T. delbrueckii presence. T. delbrueckii introduction during CM resulted in wines with increased anthocyanin content and a particular volatile profile. Isoamyl acetate, a key aroma in CM wines, was notably elevated, especially in the TdP strain. Sensory evaluations also revealed distinctions, with TdV wines displaying more pronounced aromas of red fruit, banana, and grass. Regarding MLF, T. delbrueckii presence notably enhanced performance, particularly in spontaneous MLF cases, accelerating fermentation completion. Inoculating the Oenococcus oeni strain OoVP41 also shortened MLF duration. These findings highlight the potential of Td strains to improve MLF efficiency and sensory attributes in CM wines. Employing T. delbrueckii strains strategically enables winemakers to optimize MLF and improve sensory profiles, offering an opportunity to produce higher-quality CM wines.

Cigar stacking fermentation is a key step in tobacco aroma enhancement and miscellaneous gas reduction, which both have a great influence on increasing cigar flavor and improving industrial availability. To analyze the effect of cigar tobacco leaf (CTLs) microbial community structure on volatiles during cigar stacking fermentation, this study used multi-omics technology to reveal the changes in microbial community structure and volatiles of different cigar varieties during stacking fermentation, in addition to exploring the interaction mechanism of microbiome and volatiles. The results showed that the dominant microbial compositions of different CTL varieties during stacking fermentation were similar, which included Staphylococcus, Corynebacterium 1, Aerococcus, and Aspergillus. These dominant microbes mainly affected the microbial community structure and characteristic microorganisms of CTLs through microbial interactions, thereby influencing the transformation of volatiles. Characteristic microorganisms of different CTLs varieties such as Trichothecium, Trichosporon, Thioalkalicoccus and Jeotgalicoccus, were found to posively correlate with characteristic volatiles like megastigmatrienone 4, pyrazine, tetramethyl-, geranyl acetone, and 2-undecanone, 6,10-dimethyl-, respectively. This in turn affected the aroma and sensory quality of the CTLs. This study provides theoretical support for the analysis of the mechanism of microorganisms on volatiles and aroma, and development of microbial agents during cigar stacking fermentation.

Potato virus Y (PVY) is one of the most important constraints to potato production worldwide. There is an increasing occurrence of recombinant PVY strains PVYNTN and PVYN-Wi and a decline in the incidence of the nonrecombinant PVYO. We hypothesized that this may be due to the ability of these recombinant strains to antagonize and/or outcompete PVYO in mixed infections. To deter-mine this, we investigated interactions between PVYO and three recombinant PVY strains com-mon in North America: PVYNTN, PVYN-Wi, and PVYN:O. Overall, our study showed that these inter-actions are tissue dependent. Specifically, PVYNTN, the main causal agent of potato tuber necrotic ringspot disease (PTNRD), was found to be more adaptable than PVYO, especially in potato leaves due, at least in part, to the Ny gene that confers hypersensitive resistance (HR) to PVYO. Further-more, PVYN-Wi was found to repress PVYO in potato tubers but act synergistically in potato leaves. The PVYO-induced foliage necrosis in cultivar ‘Ranger Russet’ was observed to be more severe in plants co-infected by PVYN-Wi and PVYN:O, respectively, resulting in plant death. Strikingly, this PVYO -induced necrosis was suppressed by PVYNTN in doubly infected plants. These interactions may, at least partially, explain the decreasing incidence of PVYO in United States potato produc-tion regions, especially given that many cultivars contain the Ny gene, which likely limits PVYO enabling PVYNTN and PVYN-Wi to outcompete. We also found that replication and cell-to-cell movement of these PVY strains in tubers at 4C was similar to levels at ambient temperature.

There is a major problem with the rising occurrence of highly virulent and multiply-resistant strains, including methicillin-resistant Staphylococcus aureus (MRSA), because of their difficult treatment. This study aimed to evaluate the antibacterial and antibiofilm effect of new enterocins (Ent) against potential pathogenic MRSA strains isolated from rabbits. Staphylococci (n =110) were identified with PCR and screened for methicillin/oxacillin/cefoxitin resistance (MR) using the disk diffusion method and the PBP2' Latex Agglutination Test Kit. Enzyme production, hemolysis, DNase activity, slime production, and biofilm formation were tested in MRSA strains. The susceptibility of MRSA to eight partially-purified enterocins (Ent) produced by E. faecium and E. durans strains was checked using agar spot tests. The antibiofilm activity of Ents was tested using a quantitative plate assay. Out of 14 MRSA, PBP testing confirmed MR in 8 strains. The majority of MRSA showed DNase activity and β-hemolysis. Slime production and moderate biofilm formation were observed in all strains. MRSA were susceptible to tested Ents (100–12800 AU/mL; except Ent4231). The antibiofilm effect of Ents (except Ent4231) was noted in the high range (64.9-97.0%). These results indicate that enterocins offer a promising option for the prevention and treatment of bacterial infections caused by biofilm-forming MRSA.

This study is the first report of the successful cultivation of Dinophysis norvegica isolated from Japanese coastal waters (7 of 48 isolated cells were established as the clonal cultures, 14.5%) and their toxin content and production. The strains were maintained at relatively high abundance (>2,000 cells mL-1) for more than one year when fed on Mesodinium rubrum with the addition of Teleaulax amphioxeia. At the end of the one-month incubation, the total amounts of pectenotoxin-2 (PTX2), dinophysistoxin-1 (DTX1) and okadaic acid (OA) were 132.0-375.0 ng mL-1 (n = 7), 0.7-3.6 ng mL-1 (n = 3) and trace level OA (n = 1), respectively. Similarly, the cell quota of PTX2 and DTX1 were 60.6-152.4 pg cell-1 (n = 7) and 0.5-1.2 pg cell-1 (n = 3), respectively. These data indicated that toxin production varies depending on the strains in this species. In the growth experiment, D. norvegica grew, but it was very slow first 12 days. However, after that they grew exponentially with the maximum growth rate of 0.56 divisions day-1 (during Day 24-27), reaching a maximum concentration of 3,000 cells mL-1 at the end of incubation (Day 36), suggesting that they have a long lag phase. In the toxin production, OA was not at a detectable level (≤ 0.010 ng mL-1) during the 36 days of incubation except for Day 6. The concentration of DTX1 and PTX2 showed similar patterns as described for vegetative growth, but the toxin production still increased on Day 36 (1.3 ng mL-1 and 154.7 ng mL-1 in DTX1 and PTX2, respectively). The findings of this study provide novel information on the toxin content and production in D. norvegica as well as details on the culturing and maintenance of the species.

AbstractCurrently, public health researchers are investigating several complex and multi-faced health problems that require mixed-method approaches (MMA) to clearly understand problems and design proper intervention approaches and policy formulation. Besides, a comprehensive comprehension of the complex operation of society is vital. The MMA helps in having a broader understanding of the functioning and structure of the community at large. MMA deliberately integrates qualitative and quantitative approaches carefully to build on the merits of each other and confirm that the findings of research are closer to the truth. Therefore, public health scholars must have an excellent comprehension of the MMA and its philosophical foundations for discovering the complex conditions that challenge public health nowadays. However, public health researchers do not clearly understand the theoretical underpinnings and practical application of MMA, despite its popularity in developing countries. Therefore, this article aimed to provide comprehensive evidence on the theoretical underpinnings and practical application of MMA.

Ontological premises have profound theoretical implications. This analysis posits a single, universal structure composed of two ontic entities: discrete, quantized units of space ("Planck Spheres") and a State of Absolute Nothingness (the "SOAN"). Planck Spheres and the SOAN form a physical, ultra-high (3 x N) dimensional space ("Planck Space") and a discrete 3D space (or "4D spacetime"). Together, Planck Space and 4D spacetime form a tightly integrated ((3 x N) + 3) hyperspace (the "Dual Ontology"). Critically, a one-to-one mapping and identity exists between the Planck Spheres comprising 4D spacetime and Planck Space. The Dual Ontology conjecture supports radically simplified physical explanations for three distinct areas of theoretical physics: First, the conjecture reconciles fundamental issues in Special Relativity and quantum mechanics by explaining why single and N-body quantum states (a) dynamically evolve in 4D spacetime subject to the speed of light and (b) instantaneously collapse in a physical Planck Space that is not subject to Special Relativity. Second, the temporal and physical asymmetry between the dynamic evolution of quantum states in 4D spacetime subject to Special Relativity and the instantaneous collapse of quantum states in a Planck Space where Special Relativity is inapplicable explains quantum path irreversibility and the arrow of time. Third, at or near the heat death of 4D spacetime, the instantaneous collapse of the energy content of Planck Space and the simultaneous transition of 4D spacetime's extremely widely dispersed energy content to a non-singular, generally localized volume at t = 0 explains 4D spacetime's isotropy, homogeneity, extremely high energy, pressure, and temperature, flatness, and low gravitational entropy. It also explains the horizon and fine-tuning problems.

(1) Background: Heat stress detrimentally restricted the economic in dairy farming production, therefore, this study was designed to investigate how automatic spraying method on alleviating heat stress on cows. (2) Methods: A total of 1208 multiparous dairy cows were randomly allocated into 6 barns, three barns were equipped with automatic sprinklers (SP), each barn was considered as a replicate. The other three barns were considered as control treatments (CON). Each barn was considered as a replicate. (3) Results: body temperature and milk somatic cell count were significantly decreased, while DMI, milk yield, and milk fat content significantly increased with spraying treatment. ICR was improved from 30% to 35%. Rumen fermentation was enhanced by the increased levels of total VFA, acetate, propionate, and butyrate with the spraying treatment. Rumen microbiota results showed at the phylum level, rumen Fibrobacters abundance significantly increased by the spraying treatment. At the genus level, Eubacterium, Shuttleworthia, Saccharofermentans, Lachnospira, Pseudobutyrivibrio, Selenomonas, and Succinivibrio abundance were significantly increased, while Ruminococcaceae and Rikenellaceae abundance were significantly decreased in the rumen with spraying treatment. (4) Conclusion: this study demonstrated automatic spraying modulated the rumen microbiota composition and fermentation function, and effectively meliorated side-effects of heat stress on dairy cows.

The yeast Saccharomyces cerevisiae ensures successful fermentation in winemaking, although the persistent use of commercial strains lead the loss of aroma complexity of wines. Hence, the research of indigenous S. cerevisiae with proper oenological features and well adapted to specific wine-growing area become of great interest for winemakers. Here, 206 S. cerevisiae strains were isolated from two wineries during two-year sampling campaign and bio-typed with the aim to evaluate the occurrence and persistence of S. cerevisiae wild population linked to each winery. Both wineries belonging to the same Verdicchio DOC wine area (Castelli di Jesi), and never used commercial yeasts during fermentation. Results showed a specific and varied population of S. cerevisiae in each winery without cross-contamination each other and with commercial starter strains. Moreover, inside each winery it was observed a persistent of some dominant biotypes over the time, indicating a sort of “winery-effect”. The evaluation of S. cerevisiae populations for the oenological characters showed a proper and well distinct aromatic imprinting on the resulted wines confirming the “winery effect”.

Trichoderma atroviride is a strong necrotrophic mycoparasite antagonizing and feeding on a broad range of fungal phytopathogens. It further beneficially acts on plants by enhancing growth in root and shoot and inducing systemic resistance. Volatile organic compounds (VOCs) are playing a major role in all those processes. To date, T. atroviride IMI 206040 and T. atroviride P1 are among the most frequently studied T. atroviride strains and hence are used as model organisms to study mycoparasitism and photoconidiation. However, there are no studies available, which systematically and comparatively analyzed putative differences between these strains regarding their light-dependent behavior. We therefore explored the influence of light on conidiation and the mycoparasitic interaction as well as the light-dependent production of VOCs in both strains. Our data show that in contrast to T. atroviride IMI 206040 conidiation in strain P1 is independent of light. Furthermore, significant strain-and light-dependent differences in the production of several VOCs between the two strains became evident, indicating that T. atroviride P1 could be a better candidate for plant protection than IMI 206040.

The work is devoted to the investigation of virus quasispecies evolution and diversification due to mutations, competition for host cells, and cross-reactive immune responses. The model consists of a nonlocal reaction-diffusion equation for the virus density depending on the genotype considered as a continuous variable and on time. This equation contains two integral terms corresponding to the nonlocal effects of virus interaction with host cells and with immune cells. In the model, a virus strain is represented by a localized solution concentrated around some given genotype. Emergence of new strains corresponds to a periodic wave propagating in the space of genotypes. The conditions of appearance of such waves and their dynamics are described.

Kluyveromyces marxianus is an attractive thermotolerant yeast species for ethanol production because of its ability to utilize various carbon sources as a fermentation substrate. The use of thermotolerant microbes enables performance of high-temperature ethanol fermentation, which has several advantages including reduction of cooling costs and minimization of contamination risk. To improve K. marxianus for ethanol fermentation under stress conditions, two strains, DMKU 3-1042 and DMKU 3-118, were adapted for heat resistance and toxic substances in pulp wastewater from a paper mill, respectively, resulting in the generation of KMR1042 and KMR118, respectively. Both adapted mutants exhibited clumpy clusters of cells as pseudohyphae and altered colony morphology, and their sedimentation speeds were much faster than those of the corresponding parent strains. The two mutants showed stronger tolerance to various stresses and higher performance for ethanol production than those of the corresponding parent strains at high temperatures or in the presence of toxic substances. Genome sequencing analysis revealed that both mutants had disruption of the same gene, SWI5, despite adaptation under different stress conditions, suggesting that the formation of pseudohyphae is a common strategy of K. marxianus for coping with stresses.

As one of the most appealing edible mushrooms, the quality of Flammulina velutipes can be affected by the cultivation substrate, which has an impact on the mushroom umami taste and nutrients. In our study, the effect of mushroom root fermentation broth (MRFB) based substrate on umami taste and nutrients of F. velutipes were evaluated; four proportions of MRFB was conducted (CK 0%, E1 6.7%, E2 13.4%, E3 20.1%). Results indicated that MRFB was an effective nutrient supplement for mushroom cultivation substrate. E2 and E3 showed higher crude fiber, crude fat, soluble protein, and soluble sugar. Compared to CK, the content of monosodium glutamate-like (MSG-like) amino acids, essential amino acids, total amino acids and total 5'-nucleotides in E2 and E3 were higher. The equivalent umami concentration (EUC) values in E1, E2 and E3 were 1.70, 2.43 and 1.56 times of CK, respectively. Higher umami, saltness and sweetness taste values were found in E2. Thereby, it signified that better umami and richer nutrients were achieved by using substrate with proper volume of MRFB, especially, E2 with 13.4% of MRFB. Overall, better mushroom quality could be derived from the application of MRFB in cultivation. MRFB was a favorable nutrient supplement for Flammulina velutipes cultivation substrate.

The present study has been one of the first attempts to thoroughly examine the effects of different kefir sources on fermentation characteristics, aerobic stability, and microbial communities of alfalfa silages. The effects of commercial kefir (CK) and homemade kefir culture (HK) applied with four different application doses (untreated control (CON), 5.0, 5.7, and 6.0 log cfu g-1) on wilted alfalfa and stored at an ambient temperature of 25-30 °C. After 45 days ensiling, fermentation characteristics and aerobic stability of silages were measured and bacterial diversity was investigated by 16S ribosomal RNA gene sequencing using GenomeLab™ GeXP platform. Both CK and HK accelerate more lactic acid production and reduced ammonia nitrogen concentration. Factor analysis of kefir sources suggest that the addition of kefir improves the aerobic stability of silages even the initial water soluble carbohydrate (WSC) content is inadequate via its antimicrobial effect on yeast and mould formation. Enterococcus faecium, Pediococcus pentosaceous, and Lactobacillus brevis were dominant bacterial species among the treated groups at silo opening while Lactobacillus plantarum and Lactobacillus brevis became dominant bacterial species after 7 days of aerobic exposure. In conclusion, application of kefir on alfalfa silages improves fermentation quality and aerobic stability even with low WSC content.

Polyhydroxyalkanoates (PHA) are microbial biopolyesters utilized as “green plastics”. Their production under controlled conditions resorts to bioreactors operated in different modes. Because PHA biosynthesis constitutes a multiphase process, both feeding strategy and bioreactor operation mode need smart adaptation. Traditional PHA production setups based on batch, repeated batch, fed-batch or cyclic fed-batch processes are often limited in productivity, or display insufficient controllability of polyester composition. For highly diluted substrate streams like it is the case for (agro)industrial waste streams, fed-batch enhanced by cell recycling were recently reported as a viable tool to increase volumetric productivity. As emerging trend, continuous fermentation processes in single-, two-, and multi-stage setups are reported, which bring the kinetics of both microbial growth and PHA accumulation into agreement with process engineering, and allow tailoring PHA´s molecular structure. Moreover, we currently witness an increasing number of CO2-based PHA production processes using cyanobacteria; these light-driven processes resort to photobioreactors similar to those used for microalgae cultivation, and can be operated both discontinuously and continuously. This development goes in parallel to the emerging use of methane and syngas as an abundantly available gaseous substrates, which also calls for bioreactor systems with optimized gas transfer. The review sheds light on the challenges of diverse PHA production processes in different bioreactor types and operational regimes using miscellaneous microbial production strains such as extremophilic Archaea, chemoheterotrophic eubacteria and phototrophic cyanobacteria. Particular emphasize is dedicated to the limitations and promises of different bioreactor-strain combinations, and to efforts devoted to upscaling these processes to industrially relevant scales.

Catalase can decompose hydrogen peroxide in plants under controlled and stressed condi-tions. In a stressed environment, the increase in hydrogen peroxide production makes catalase activity a major defense system for plant growth. The current study was conducted to evaluate the catalase activity of pre-isolated and identified bacterial strains Bacillus aryabhattai (AN30), Bacillus megaterium (AN24), Bacillus megaterium (AN31) and Bacillus megaterium (AN35) and their potential for rice seedlings growth promotion. These strains were characterized for quantita-tive catalase, urease, siderophores, and exopolysaccharides production using LB media. Sub-sequently, the effectiveness of these strains was checked by quantifying the catalase activity in the rhizosphere of rice seedlings. Furthermore, the catalase activity was quantified in the roots and shoots of rice seedlings. Plant growth parameters were also studied in a growth room trial. The strain AN24 showed the maximum catalase activity (1.36 mol cm-1), urease ac-tivity (1.35 mol cm 1), exopolysaccharides (4.20 µg mL-1), and siderophores (2.32%) production in LB media. All tested strains showed significantly higher catalase activity in soil compared to control. Among sole application, strain AN24 showed better results; however, consortium application of strains AN24+AN30+AN35+AN31 showed the maximum increase in catalase ac-tivity in the rhizosphere of rice seedlings. Moreover, consortium application of AN24+AN30+AN31+AN35 resulted in the highest increase in catalase activity, dry biomass of shoots and roots, shoot and root length of rice seedlings. The results showed that a consortium of these Bacillus strains with catalase activity has greater potential to enhance an antioxidant defense system and growth promotion of rice seedlings. However, further experimentation under natural conditions is required before using these strains as potential bioinoculants for improving the rice growth and yield.

A total of thirty-two aerobic spore former strains were isolated from intestinal samples of healthy children and analyzed for their hemolytic and antibiotic-resistance activities. Four strains selected as non-hemolytic and sensitive to all antibiotics recommended as relevant by regulatory agencies were short-listed and evaluated for their in silico and in vitro probiotic potentials. The four selected strains were assigned to the Bacillus velenzensis (MV4 and MV11), B. subtilis (MV24) and Priestia megaterium (formerly Bacillus megaterium) (MV30) species. A genomic analysis indicated that MV4, MV11 and MV24 contained a homolog of the gene coding for the fibrinolytic enzyme nattokinase while only MV30 encoded for a glutamic acid decarboxylase essential to synthesize the neurotransmitter GABA. All four strains showed a strong antioxidant activity, formed biofilm and produced/secreted quorum sensing peptides able to induce a cytoprotective stress response in a model of human intestinal (HT29) cells. Altogether, genomic and physiological data indicate that the analyzed strains do not pose safety concerns and have in vitro probiotic potentials.

Group B Streptococcus (GBS), a commensal in the body, causes a wide range of infectious diseases. The colonisation levels of GBS and its resistance profile to antibiotics provide important information useful for orienting prevention strategies. There is little data available on the subject with determination of resistance phenotypes in Cameroon. We therefore aimed to determine the prevalence of colonization, antibiotic resistance, including patterns of inducible resistance to clindamycin of GBS in Yaounde. To achieve this goal, a prospective cross-sectional study with an analytical component was carried out from the 28th June to the 29th August 2020 at the BIOSANTE laboratory and the Yaounde Gynaeco-Obstetrics and Paediatrics hospital. Vaginal swabs and urine were collected on 163 women. This samples were analysed using 5% defibrinated sheep blood agar and chocolate plus polyvitex agar. The isolates were identified using the morphology of the colony, Gram staining, haemolysis, catalase test and latex grouping test. Antibiotic susceptibility testing was done by disk diffusion method following the recommendations of the ACFSM 2019. The double disk diffusion method was used to identify isolates with clindamycin inducible resistance. Our data was analysed by the software SPSS version 2.1. The results obtained showed that the global prevalence of colonization by GBS was 37% (57/163), 40.35% in non-pregnant women and 59.65% in pregnant women. Pregnancy (P-value = 0.019) and gestational age (P-value = 0.025) constituted the risk factors of maternal colonization by GBS. In addition, the strains of GBS were resistant to all antibiotics tested. A D test showcased that 64.7% of GBS were resistant in a constitutive manner to clindamycin. We also note the presence of M phenotypes. As a whole, our results demonstrate that the rate of GBS colonization in this study was similar or higher than those in the previous report in Cameroon. All this indicates that attention should be paid to this bacterium in the monitoring of antimicrobial resistance and in the care of pregnant women and newborns.

Infectious bronchitis virus (IBV) is a highly variable RNA virus that affects chickens worldwide. Due to its inherited tendency to suffer point mutations and recombination events during viral replication, emergent IBV strains have been linked to nephropathogenic and reproductive disease that are more severe than the typical respiratory disease, leading, in some cases, to mortality, severe production losses, and/or unsuccessful vaccination. QX and DMV/1639 strains are examples of the above-mentioned IBV evolutionary pathway and clinical outcome. In this study, our purpose was to systematically compare whole genomes of QX and DMV strains looking at each IBV gene individually. Phylogenetic analyses and amino acid site searches were performed in datasets obtained from GenBank accounting for all IBV genes and using our own relevant sequences as a basis. The QX dataset studied is more genetically diverse than the DMV dataset, partially due to the greater epidemiological diversity within the five QX strains used as a basis compared to the four DMV strains from our study. Historically, QX strains have emerged and spread earlier than DMV strains in Europe and Asia. Consequently, there are more QX sequences deposited in GenBank than DMV strains, assisting in the identification of a larger pool of QX strains. It is likely that a similar evolutionary pattern will be observed among DMV strains as they develop and spread in North America.

The genus Rhodotorula includes basidiomycetous oleaginous yeast species. R. babjevae can produce compounds of biotechnological interest such as lipids, carotenoids and biosurfactants from low value substrates such as lignocellulose hydrolysate. High-quality genome assemblies are needed to develop genetic tools and to understand fungal evolution and genetics. Here, we combined short- and long-read sequencing to resolve the genomes of two R. babjevae strains, CBS 7808 (type strain) and DBVPG 8058 at chromosomal level. Both genomes have a size of 21 Mbp and a GC content of 68.2%. Allele frequency analysis indicated tetraploidy in both strains. They harbor 21 putative chromosomes with sizes ranging from 0.4 to 2.4 Mb. In both assemblies, the mitochondrial genome was recovered in a single contig, which shared 97% pairwise identity. The pairwise identity between the majority of chromosomes ranges from 82% to 87%. We found indications for strain-specific extrachromosomal endogenous DNA. 7,591 protein-coding genes and 7,607 associated transcripts were annotated in CBS 7808 and 7,481 protein-coding genes and 7,516 associated transcripts in DBVPG 8058. CBS 7808 has accumulated a higher number of tandem duplications than DBVPG 8058. We identified large translocation events between putative chromosomes and a high genetic divergence between the two strains.

The bloom-forming toxic dinoflagellate Alexandrium catenella was first detected in Southern Chile (39.5–55°S) 50 years ago and is responsible for most of the area’s cases of paralytic shellfish poisoning (PSP). Given the complex life history of A. catenella, which includes benthic sexual cysts, in this study we examined the potential link between latitude, toxicity, and sexual compatibility. Nine clones isolated from Chilean Patagonia were used in self- and out-crosses in all possible combinations (n=45). The effect of latitude on toxicity, reproductive success indexes, and cyst production was also determined. Although the toxin profiles were similar for all strains, consisting of C1, C2, GTX4, GTX1, GTX3, and NeoSTX, a latitudinal gradient was determined for their proportions (%) and content per cell (pg cell⁻¹), with the more toxic strains occurring in the north (−40.6°S). Reproductive success also showed a latitudinal tendency and was lower in the north. None of the self-crosses yielded resting cysts. Rather, the production of resting cysts was highest in pairings of clones separated by distances of 1000–1650km. Our results contribute to a better understanding of PSP outbreaks in the region and demonstrate the importance of resting cysts in fueling new toxic events. They also provide additional evidence that the introduction of strains from neighboring regions is a cause for concern.

Brucella abortus is a facultative intracellular Gram-negative bacterium that causes chronic persistent infections in humans and livestock. In this study, conventional bio-typing, multiple-locus variable-number tandem repeat analysis (MLVA), and whole-genome sequencing-single-nucleotide polymorphism (WGS-SNP) were used to investigate the molecular epidemiology characteristics of Brucella abortus strains in China and their relationships to world lineages. A total of 100 strains were collected from 1953 to 2013, suggesting that B. abortus circulated in China in the past five decades. Moreover, most strains were mainly distributed in the northwest area. During this period, seven biovars were found, indicating that B. abortus displayed a high diversity of biovars and that the main epidemic areas are in the northern provinces. Strains have high genetic diversity, and bruce07 is the most helpful locus for genotyping of this population. Moreover, 17 MLVA-11 genotypes were found; 13 of them are of known genotypes and four are unassigned genotypes, indicating that B. abortus in this study had several geographic origins. Still, strains from unassigned genotypes may originate from China. Many shared MLVA-16 genotypes were observed in strains from the same provinces in Northern China, which confirmed a brucellosis outbreak within northern regions. WGS-SNP analysis showed that eight Chinese strains formed a ladder-like phylogram (C. Ⅶ) with strains from nine countries, including Uganda, Iraq, Russia, Georgia, Spain, Italy, Egypt, Mongolia, and China; strains were introduced to these countries from a single source. Notably, Chinese strains were closely related to strains from Russia and Mongolia; frequent animal (cattle) trade and exchanges may promote this process. We will provide new and insightful information for strength surveillance and control of B. abortus brucellosis in China.

We derive a quantum speed limit for unitary evolution for the case of mixed quantum states using the stronger uncertainty relation for mixed quantum states. This bound can be optimized over different choices of Hermitian operators for a better bound. We illustrate this with some examples and show its better performance with respect to three existing bounds for mixed quantum states.

Polypropylene (PP) is commonly used in the polymer processing industry. Its feature is a good chemical resistance, it is non-toxic and not harmful to people. Polypropylene is widely used in the manufacturing of food packages, laboratory and medical equipment, electric cables insulators, automotive parts, toys and furniture. The application of this material enables us to obtain complicated shapes of big surfaces. It is also highly recyclable. Polylactide (PLA) is a product of the so called “green chemistry” because it is obtained from the renewable raw materials (biomass). As its price is higher compared to the traditional polymers, its repeated use (recycling) is of high interest. Due to the same reason (cost) the mixtures of PLA with other polymers are applied. In this paper the results of testing the PP/PLA mixtures of various mass ratio of the components are presented. Technical terms of preparing and processing such mixtures have been practically tested. Also the mechanical strength was determined based on the static tensile tests. The results showed that those mixtures can be repeatedly processed. Even five-times reprocessing does not cause essential drop of mechanical properties

Triple-negative breast cancer (TNBC) is a subtype of breast cancer characterized by the lack of estrogen receptors (ER), progesterone receptors, and HER-2 receptors. Thus, TNBC tumours do not benefit from the current therapies targeting ER or HER-2. Therefore, there is an urgent need to develop novel treatment for this subtype of breast cancer. Marijuana is a common name given to Cannabis plants, a group of plants in the Cannabis genus of the Cannabaceae family. Cannabis plants are among the oldest cultivated crops, traced back at least 12,000 years and are well known for their multi-purpose usage, including medicinal purposes. The main active compounds extracted from Cannabis plants are 21-carbon-containing terpenophenolics, which are referred to as phytocannabinoids. Of these, the tetrahydrocannabinol (THC) group contains highly potent cannabinoids, including delta-9-tetrahydrocannabinol (∆9-THC) and delta-8-tetrahydrocannabinol (∆8-THC), which are the most abundant THCs and are largely responsible for psychological and physiological effects of marijuana. The use of Cannabis plants for medicinal purposes was first recorded in 2337 BC in China, where Cannabis plants were used to treat pains, rheumatism, and gout. Recently, several cannabinoids have been approved for a number of treatments, one of which is the treatment of nausea and vomiting caused by chemotherapy in cancer patients. Furthermore, increasing evidence shows that cannabinoids not only attenuate side effects due to cancer treatment, but might also potentially possess direct antitumor effects in several cancer types, including breast cancer. However, anti-tumour activity of marijuana has been variable in different studies and even promoted tumour growth in some cases. In addition, the mechanisms of cannabinoid action in cancer remain unclear. This review summarizes evidence about the mixed actions of cannabinoids in cancer in general and triple-negative breast cancer in particular.

At present European buildings typically consume two to five times more energy than predicted at the design stage. An important cause of this performance gap is the discrepancies between the design specification and the As-Built condition. Such discrepancies are mainly due to the gaps in knowhow between design, production and construction professionals. Design is more and more contained into a virtual environment and loses touch with the physical production and construction sites. As the construction sector enters the Industry 4.0 era, Building Information Modelling (BIM) based Mixed Reality can intertwine virtual and real worlds to bridge the knowhow gaps.

The aim of this work is to present a design approach of a Shock-Induced Combustion Ramjet (Shcramjet) inlet, and present its optimization in terms of the flow and geometrical parameters. The flow properties of mixed compression type inlet of a Shcramjet are examined using analytical and numerical techniques. The geometries obtained with variations in the wedge angles, length, height and cowl lip positions are used to study the flow characteristics of inlet, identifying bow shock temperature ratio as the optimization parameter. The two-dimensional geometries of two-shock and three-shock inlet models designed analytically for shock-on-lip condition at Mach 12.5 and an altitude of 32.5 km are numerically simulated in OpenFOAM CFD Toolbox. A density based compressible CFD solver based on central upwind schemes of Kurganov and Tadmore is used to solve 2D inviscid Euler equations. The inlet total pressure recovery is expressed as a function of temperature ratios of compression shocks, and is found to have a maxima at a bow shock temperature ratio at the design condition when the bow and external shocks have equal strengths. The effect of considering the flow to be calorically imperfect is studied numerically, and the deviation from the analytical design is presented.

The aim of this study was to assess the effects of supplementation of inorganic sources of manganese (MnO, MnSO4), zinc (ZnO, ZnSO4) and copper (CuSO4) at different levels (0.06%DM for Mn, 0.05%DM for Zn; and 0.01 and 0.05%DM for Cu) on in vitro rumen fermentation, solubility and bioavailability. Fermentation activity was measured by total gas production (TGP) and dry matter degradability after 70hrs of fermentation (dDM%). Trace-mineral (TM) solubility was estimated by the TM concentration in the supernatant of the final fermentation medium (SOL) and TM bioavailability by the TM concentration in a bacterial enriched fraction (BACT). Mn (regardless of source) and ZnO tended (p&lt;0.10) to decrease, while Cu showed no significant effect on TGP. The addition of inorganic Mn and of ZnO tended (p&lt;0.10) to decrease, ZnSO4 tended to increase (p&lt;0.10), whilst Cu showed no effect on dDM%. Concerning solubility, Mn (MnO and MnSO4), ZnSO4 and CuSO4 significantly (p&lt;0.05, p&lt;0.001 and p&lt;0.01) increased, while ZnO did not affect TM content of the SOL. These results indicate that MnSO4, ZnSO4 and CuSO4 are highly soluble, MnO is quite soluble, while ZnO has a low solubility in the rumen. Based on the TM content of BACT, MnO, MnS4 and CuSO4 have a high bioavailability, while ZnO is poorly assimilated by rumen bacteria. However, the lack of clear inhibition or improvement of fermentations suggests that the rumen microbiota has low requirement for TM supplementation.

Limonene is a valuable monoterpenoid that with diverse applications. Microbial biosynthesis offers a sustainable alternative to plant-based extraction, which is prone to hinderance by climate change and acreage. In the present study, the effects on limonene production of carbon source, inoculum, working volume, temperature, pH, dodecane coverage, and media were investigated with engineered strains of Rhodosporidium toruloides. Through single-factor and orthogonal experiments, the limonene titer was improved from 52.2 to 358.1 mg/L on 250-mL flask, a 586% increase. The optimized medium was MM, with a working volume of 50 mL, inoculum OD600=0.6, 22 °C, pH 6, and dodecane coverage (v/v) of 20%. The results here demonstrated that the R. toruloides could be explored as an efficient limonene producer and facilitate other monoterpenoids biosynthesis with R. toruloides.

Single cell proteins are the dead dried cells of microorganisms or purified protein isolated from microorganism’s cell culture, used as a food supplement to humans’ food and animals feed. World suffer from malnutrition particularly developing countries, due to rapid increase in population, increased the demand for protein and nutrients requirement. Bacteria is potential microorganism for SCP production due to high protein content, fast generation time, bio-active secondary metabolites production and can grow on various substrates. Actinobacteria species and strain have capability to produce biological active compounds, produced about two-thirds of antibiotics available in the market, actively used as antibiotics, antiprotozoal, antifungal, antiviral, anticancer, anticholesterol, antihelminth and immunosuppressant. Actinomycetes can be used as probiotic as well as single cell protein that will deal with antibiotic as well as protein source.

The production of lycopene from different substrates by Blakeslea trispora in fermentation was investigated. Lycopene productions from 4 and 6 % glucose (pH 6.5) in shake flask fermentation were 77.7 and 28.1 mg L-1. A maximum lycopene concentration of 944.8 mg L-1 was detected with 4 % glucose supplemented with 1.0 % sunflower oil in fermentor studies. Zygospores of B. trispora are the morphological forms, which are responsible for the production of the lycopene. The highest level of zygospores was correlated with the highest amount of intracellular lycopene in the total biomass dry weight. The media containing only orange peel (1 %) gave a 4.9 mg L-1 lycopene production in a fermentor. The presence of oils as substrates resulted in enhanced mold growth and subsequent higher lycopene production. Substrates containing linoleic acid compounds led to high lycopene production. The data showed that the biosynthesis of lycopene starts in most cases simultaneously in the early growth phase even in trace amounts and the amount of lycopene formation increased continuously from 2 to 7 days.

Rhizobia are biological nitrogen fixing non-pathogenic micro-organisms that make inert nitrogen available to legumes establishing symbiosis and living within their nodules and promoting their growth. Livestock farmers use antibiotics without prescriptions to prevent losses, death and economic wastes, resulting in indiscriminate release of excess, un-utilized antibiotics into the environment causing detrimental effects to soil microbes particularly rhizobia.This study evaluates the susceptibility pattern of selected rhizobia strains. The intrinsic antibiotic resistance (IAR) ability of common rhizobia strains including FA3, B574, RS15, USDA136, USDA9032, R25B, 532C, CC511, RANI22, RAUG1 were investigated. The antibiotics used included Ceftazamine , Florfenicol , Cefpodoxime, Sulphamethoxazole, Ciprofloxacin, Gentamycin , Kanamycin, Ertapenem, Tetracycline , Meropenem and Carbapenem. Results showed different resistance patterns among isolates where high resistance was shown against Carbapenem and least resistance was shown against Ciprofloxacin (40mm), Tetracycline (33mm), kanamycin (45mm) and Gentamycin (50mm). Rani 22 showed most resistant. Most strains showed no resistance to these antibiotics, but it is significant to note that those with resistance can be used for the recovery of antibiotics contaminated sites and may provide potential solutions for futuristic problems associated with AMR.This study suggests that using a consortium of symbiotic rhizobia strains is a strategy to achieve optimal yields in antibiotic contaminated fields as the strains showed differences in their susceptibility pattern. Also, the antibiotics produced or contained active compounds and substances that are detrimental for the growth of rhizobia and may therefore be a hindrance to these plant growth promoting bacteria when they are applied to aid plant growth used during agricultural practises.

Clostridioides (Clostridium) difficile is an enteric pathogen of neonatal pigs with zoonotic transmission remaining a significant problem in humans, including nosocomial resurgence, following oral administration of broad-spectrum antibiotics. To date, the immune response to C. difficile has mostly focused on neutrophils and cytokine/chemokines, particularly in human infection. We show that porcine monocytes respond to C. difficile differently compared with many other bacterial infections. Infection of porcine monocytes with human C. difficile strains CD630 (Ribotype 078) or R20291 (Ribotype 027) for 3 or 24h post-infection (pi) resulted in a lack of oxidative burst or nitrite ion production when compared to uninfected controls (P >0.05). The survival dynamics of both CD630 and R20291 in monocytes were similar with intracellular bacterial numbers being similar at 3h pi and 24h pi (P >0.05). However, we show that porcine monocytes entrap C. difficile via extracellular DNA traps. This process began as early as 3h pi and at 24h pi the nuclei appeared to be depleted of DNA, although extracellular DNA was associated with the cell membrane. Our preliminary study also suggests that entrapment of C. difficile by extracellular DNA may occur via a process of monocyte etosis.

Chondroitin sulfate is a glycosaminoglycan that has gained widespread use in nutraceuticals and pharmaceuticals, mainly for treating osteoarthritis. Traditionally, it has been extracted from animal cartilage but recently, biotechnological processes have emerged as a commercial alternative to avoid the risk of viral or prion contamination and offer a vegan-friendly source. Typically, these methods involve producing the chondroitin backbone using pathogenic bacteria and then modifying it enzymatically through the action of sulfotransferases. Despite the challenges of expressing active sulfotransferases in bacteria, the use of eukaryotic microorganisms is still limited to a few works using Pichia pastoris. To create a safer and efficient biotechnological platform, we have constructed a biosynthetic pathway for chondroitin production in S. cerevisiae. Furthermore, as genome-scale models are valuable tools for identifying novel targets for metabolic engineering, a stoichiometric model of chondroitin-producing S. cerevisiae has been developed and used in optimization algorithms. Our research has yielded several novel targets, such as uridine diphosphate(UDP)-N-acetylglucosamine pyrophosphorylase (QRI1), glucosamine-6-phosphate acetyltransferase (GNA1) or N-acetylglucosamine-phosphate mutase (PCM1) overexpression that might enhance chondroitin production, which will guide future experimental research to develop more efficient host organisms for the biotechnological production process.

Colletotrichum lini is a flax fungal pathogen, comprising differently virulent strains. In this work, we sequenced the genomes of three C. lini strains of different virulence: high virulent #390-1, medium virulent #757, and low virulent #771. We obtained more than 100× genome coverage with Oxford Nanopore Technologies reads and more than 50× genome coverage with Illumina data. To construct high-quality genomes, several assembly strategies were tested. The final assemblies were obtained using the Canu – Racon ×2 – Medaka – Polca scheme and were 54.0-55.3 Mb in length, contained 26-32 contigs, had an N50 > 5 Mb and the BUSCO completeness > 96%. Comparative analysis of the constructed genomes showed high similarity between mitochondrial and nuclear genomes. However, a rearrangement event and the loss of a 0.9 Mb contig were revealed. The candidate effectors were predicted in sets of proteins received after genome annotation. The obtained genome sequences of C. lini strains extend the knowledge on the genetic diversity of Colletotrichum species and are the basis for establishing molecular mechanisms of pathogenicity of the flax pathogen and effective control of flax anthracnose.

Pseudomonas aeruginosa is an adaptable bacterial pathogen that infects various organs, including the respiratory tract, vascular system, urinary tract, and central nervous system leading to high morbidity and mortality. Our primary focus of this study was to characterize P. aeruginosa clinical strains on the basis of pigment color production, determine its association to multidrug resistance behavior and ability to form biofilm. We identified yellow (30.1%), green (39.8%) and no pigment (30.1%) producing strains from a total of 143 clinical isolates. Yellow pigment producing strains presented significant resistance to a class of antibiotics including β-lactam (91.5%), aminoglycosides (70.5%), and carbapenems (51.9%) compared to green and non-pigmented strains. Importantly, 16.3% of yellow pigment producing strains was resistant to colistin where only 2.3% of non-pigmented and 1.8% of green pigmented strains were resistant to this agent. Moreover, yellow pigment producing strain were frequent producers of β-lactamase group of enzymes, ESBL (55.6%), MBL (55.6%), and AmpC (50%) and displayed higher frequency of efflux positive group (64.2%) compared to green (7.14%) and non-pigmented one (28.5%). Notably, green pigment producing strains when compared to non-pigmented groups also displayed antibiotic susceptibility behavior similar to yellow pigment producing strains. Although yellow pigment producing strains were strong biofilm producers, no significant association was identified between pigment and biofilm formation. Among pigmented and non-pigmented strains, majority of yellow pigment producing strains have shown MIC levels greater than the green and non-pigmented strains. Our study has demonstrated the impact of pigment coloration on susceptibility to antimicrobial agents where yellow pigment producing strains represent considerably a serious problem as due to lack of alternative agents against such transformed strain may collectively be associated with multidrug resistance development.

Genome-scale engineering and custom synthetic genomes are reshaping the next generation of industrial yeast strains. The Cre-recombinase mediated chromosomal rearrangement mechanism of designer synthetic Saccharomyces cerevisiae chromosomes, known as SCRaMbLE, is a powerful tool which allows rapid genome evolution upon command. This system is able to generate millions of novel genomes with potential valuable phenotypes, but the excessive loss of essential genes often results in poor growth or even the death of cells with useful phenotypes. In this study we expanded the versatility of SCRaMbLE to industrial strains, and evaluated different control measures to optimise genomic rearrangement, whilst limiting cell death. To achieve this, we have developed RED (Rapid Evolution Detection), a simple colorimetric plate-assay procedure to rapidly quantify the degree of genomic rearrangements within a post-SCRaMbLE yeast population. RED-enabled semi-synthetic strains were mated with haploid progeny of industrial yeast strains to produce stress tolerant heterozygous diploid strains. Analysis of these heterozygous strains with the RED-assay, genome sequencing and custom bioinformatics scripts demonstrated a correlation between RED-assay frequencies and physical genomic rearrangements. Here we show that RED is a fast and effective method to evaluate optimal SCRaMbLE induction times of different Cre-recombinse expression systems for the development of industrial strains.

Nitraria tangutorum Bobrov is a berry shrub with white flowers and red fruits, which grows in the deserts in the Tibetan Plateau, Mongolia, and western China. Its fruit N. tangutorum fruit (NTF) contains various bioactive compounds, with anti-fatigue, anti-inflammatory, and neuroprotective functions. However, the high saccharide content of NTF makes it unsuitable for diabetic patients. In this study, we fermented NTF to obtain N. tangutorum fermented juice (NTJ), and N. tangutorum fermented residue (NTR), which are suitable for diabetics to consume. We characterized the bioactive compounds in NTF, NTR, and NTJ, and found that fermentation increased the diversity of bioactive compounds, and greatly reduced sucrose, glucose, and fructose content while generating trehalose, which has the potential to manage blood glucose levels. Further, NTJ displays anti-diabetic potential due to various compounds anti-diabetic properties. This study provides a basis for further clinical research on NTJ's anti-diabetic function in humans.

The study focuses on the impact of new methods for inhibiting malolactic fermentation in white wines on their analytical and sensory properties. Enological preparations with different mecha-nisms of effect were tested: fumaric acid, chitosan, Estaan (a preparation based on tannin inhibi-tion), medium-chain fatty acids (MCFA), sulphur dioxide and a control variant where malolactic fermentation (MLF) was performed. The samples underwent analysis through HPLC (high-performance liquid chromatography) to determine concentrations of malic and lactic acid, as well as biogenic amines. GC (gas chromatography) analysis was used to monitor volatile sub-stances, alongside sensory evaluation. The study demonstrated a significant influence of indi-vidual enological preparations on the aromatic profile of the examined wines. The SO2 and MCFA variants exhibited the highest concentrations of volatile substances within the esters group, spe-cifically isoamyl acetate, 1-hexyl acetate and phenylethyl acetate. Conversely, the fumaric acid and Estaan variants displayed the lowest concentrations of these esters. The most notable disparities were observed in acetoin concentration, with the MCFA variant exhibiting the lowest values. Additionally, the chitosan variant showed higher concentrations of putrescine and spermidine compared to the MCFA and fumaric acid variants, which presented the lowest levels.

Traditional grain vinegar is fermented using multiple acetic acid bacteria (AAB) at various temperatures. A single AAB showed high acid–producing ability at 30 °C with a 5% alcohol concentration and an initial pH adjusted to 4.0. Multiple AAB were similar to single AAB, but the optimal initial pH was 3.0. Acid production ability according to the type of AAB was higher in multiple AAB than that for single AAB. That is, using multiple AAB helped to increase the titratable acidity of traditional grain vinegar. In addition, increasing the titratable acidity and content of volatile flavor compounds was advantageous when two, rather than four, AAB types were mixed and used. Titratable acidity was high at medium temperatures (30 °C), but volatile flavor compounds increased at low temperatures (20 °C) under multiple AAB. A 16S rDNA–based microbiome taxonomic profiling analysis identified differences in beta diversity due to multiple AAB and fermentation temperatures. In particular, beta diversity analysis revealed a specific pattern when a mixture of Acetobacter ascedens GV–8 and Acetobacter pasteurianus GV–22 were fermented at low temperature (20 °C). Therefore, we propose the application of multiple AAB with acidic and flavor–producing properties in traditional grain vinegar.

Lactobacillus plantarum has been observed to play a crucial role in shaping the sensory properties of chili sauce. However, the specific taste-active metabolites responsible for the desirable flavor profile of chili sauce remain inadequately characterized. This study employed a combination of metabolomics and web-based computational tools analysis to investigate the dynamic changes in taste-active metabolites during chili sauce fermentation. Initially, metabolites were rapidly annotated using a feature-based molecular network, leading to the tentative annotation of 206 metabolites, of which a significant proportion had not been previously reported. Subsequently, the VirtualTaste tool identified dihydrosphingosine, lactic acid, isoleucine, phytosphingosine, and gluconic acid as potential taste markers for quality control. Finally, pathway enrichment analysis revealed that these components were primarily associated with amino acid tRNA, phenylalanine, tyrosine, and tryptophan biosynthesis, as well as sphingolipid metabolism. This study provides valuable insights into the mechanisms underlying the formation of the distinctive flavor of chili sauce.

Several red seaweeds have shown to inhibit enteric CH₄ production; however, adaptation of fermentation parameters to their presence is not well understood. The objective of this study was to examine the effect of three red seaweeds (Asparargopsis taxiformis, Mazzaella japonica, Palmaria mollis) on in vitro fermentation, CH₄ production, and adaptation using the rumen simulation technique (RUSITEC). The experiment was conducted as a completely randomized design with four treatments, duplicated in two identical RUSITEC apparatus equipped with eight fermenter vessels each. The four treatments included the control (barley straw and barley silage) and the three red seaweeds added to the control diet at 2% diet DM. The experimental period was divided into four phases including a baseline phase (d 0-7; no seaweed included), adaptation phase (d 8-11; seaweed included in treatment vessels), intermediate phase (d 12-16) and a stable phase (d 17-21). The digestibility of organic matter (P = 0.04) and neutral detergent fibre (P = 0.05) was decreased by A. taxiformis during the adaptation phase, but returned to control levels in the stable phase. A. taxiformis supplementation resulted in a decrease (P < 0.001) in molar proportions of acetate, propionate and total volatile fatty acid (VFA) production, with an increase in molar proportions of butyrate, caproate, and valerate; the other seaweeds had no effect (P > 0.05) on molar proportions or production of individual VFA. A. taxiformis was the only seaweed to suppress CH₄ production (P < 0.001), with the suppressive effect increasing (P < 0.001) across phases. Similarly, A. taxiformis increased (P < 0.001) the production of hydrogen (H₂, %, mL/d) across the adaptation, intermediate and stable phases, with the intermediate and stable phases having greater H₂ production than the adaptation phase. In conclusion, M. japonica and P. mollis did not impact rumen fermentation or inhibit CH₄ production within the RUSITEC. In contrast, we conclude that A. taxiformis is an effective CH₄ inhibitor and its introduction to the ruminal environment requires a period of adaptation; however, the large magnitude of CH₄ suppression by A. taxiformis inhibits VFA synthesis, which may restrict production performance in vivo.

More than 800 million tons of fish are utilizing in a year and 25-30% become waste. The waste amount is beneficial source for extraction of collagen. But procedure of extraction is still to be optimized. The current study was designed to extract collagen from fish through fish waste fermentation under various conditions. For collagen extraction we did lactic acid fermentation in which yogurt and Dough bacteria was added with fish sample and placed it in incubator at 30ºC for one month. In yogurt and dough culture have at least 10 type of lactic acid bacterial species. After every week we check PH of each sample and take soup of that sample. After centrifugation of that sample’s TCA (Trichloroacetic acid) precipitation was done of yogurt and dough sample and kept at -20ºC. then did SDS-PAGE) using 6% resolving and 5% stacking gel of already stored sample. Then HaCaT cells (1 × 10⁴ cells/well) were cultured in 96-well flat-bottom culture plates and treated with appropriate doses of FFCP (fermented fish collagen peptide) for 24 and 48 hours.The SDS-PAGE revealed that the collagen protein of fish had doublet pattern for α1 and α2 chains at corresponding to 145 kDa and 132 kDa respectively. The density for α1 twice as compared to α2. Our result agrees that The fish collagen consists mostly of α-chain as well as little amount of inter and intra molecular cross-linked components of α-chains; b (dimmer) and c (trimer). Different biochemical tests were done for identification of lactic acid bacteria catalase positive, citrate positive, urease negative. Our fermented fish collagen and peptides mixture were test for cellular cytotoxicity and proliferative effect on HaCaT cells. Current result shows that fermented extracted collagen are nontoxic and induce the proliferation of HaCaT cells. Current study is supported by various studies that revealed the medical application of fish extracted collagen as underline.

Tempeh is an Indonesian traditional food, made from white soybean seeds by fermentation with Rhizopus mold. Soybean seeds content high isoflavone glycosides. Isoflavones can act as phytoestrogen, antioxidant and improves memory. Isoflavone glycosides are poorly absorbed in the human body, but the aglycone isoflavones absorbed quickly. Rhizopus mold hydrolize isoflavone glycosides to be aglycone isoflavones during tempeh production. This research was aimed to know the best room temperature and time of fermentation for making soybean tempeh with high content of isoflavones. After rinsing and boiling, the soybean seeds than fermented with Rhizopus oligosporus. Three conditions were applied: (a) ambient temperature (27-32oC) without air circulation, (b) 27±0.5 oC, and (c) 30±0.5 oC both with air circulations. Inner temperature of tempeh was recorded hourly. Total isoflavones were measured with UV spectrophotometer every 6 hours. Based on this study, fermentation at condition a caused the tempeh too hot and rotted quickly. Fermentation at condition b and c produced tempeh with good quality. Tempeh ripening was reached between 31-32 hours with inner temperature 32-33 ^oC. Fermentation for 72 hours at condition b was chosen because its high total isoflavones content (0.089% w/w), but decrease about 20% compare to soybean seeds (0.112% w/w).

Our study investigated the innate immune response to Toxoplasma gondii infection by assessing microglial phenotypic changes and sickness behavior as inflammatory response markers post-ocular tachyzoite instillation. Disease progression in Swiss albino mice was compared with documented outcomes in BALB/c mice using identical ocular route and parasite burden (2x105 tachyzoites), with saline as control. Contrary to expectations, Swiss albino mice displayed rapid, lethal disease progression, marked by pronounced sickness behaviors and mortality within 11-12 days post-infection, while survivors showed no apparent signs of infection. Comparative analysis revealed T. gondii-infected BALB/c mice exhibited reduced avoidance of feline odors, while in-fected Swiss albino mice showed enhanced avoidance responses. There was a significant increase in microglial cells in the dentate gyrus molecular layer of infected Swiss albino mice compared to BALB/c mice and their respective controls. Hierarchical cluster and discriminant analyses iden-tified three microglial morphological clusters, differentially affected by T. gondii infection across strains. BALB/c mice exhibited increased microglial branching and complexity, while Swiss al-bino mice showed reduced shrunk microglial arbors, diminishing morphological complexity. These findings highlight strain-specific differences in disease progression and inflammatory reg-ulation, indicating lineage-specific mechanisms in inflammatory responses, tolerance, and re-sistance. These elements are critical in devising control measures for toxoplasmosis.

With the continuous improvement and observable benefits of electric vehicles (EVs), major logistic companies are introducing more EVs into their conventional fleets. This gives rise to a new type of vehicle routing problem with mixed vehicles, where heterogeneous internal combustion vehicles (ICVs) and electric vehicles are considered in route planning. In addition, certain deliveries that are not efficient on any type of vehicles, are outsourced to third-party common carriers. In this paper, we define this problem as a mixed vehicle routing problem with common carriers (MVRPC). The objective of such problems is to minimize the transportation costs by considering routes with ICVs and EVs, the possibility of visiting recharging stations, outsourcing options, and drivers’ layover regulations. This variant of the vehicle routing problems has many practical applications, particularly in the design of long-haul transportation and last-mile delivery services. Effective MVRPC solutions play a key role in promoting the going Green image and optimally allocating resources. The problem has received limited attention in the literature likely because addressing all the needed aspects is especially challenging. To solve the large-scale problem, we develop a branch-and-cut pricing framework that relies on strong cuts and customized labeling algorithms. Numerical experiments highlight the effectiveness of our algorithm. This success can be attributed to tailored critical resources, dynamically bounded bidirectional labeling procedures, strong dominance criteria, and implementation strategies.

(1) Background: Mixed Reality is a technology that has gained attention due to its unique capabilities for accessing and visualizing information, becoming a valuable tool for medicine, particularly for the operating room and surgical learning; (2) Methods: A systematic review was conducted following the PRISMA guidelines to answer the research questions established using the PICO framework; (3) Results: Although implementation of Mixed Reality applications in the operations room presents some challenges, when used appropriately, it can yield remarkable results. It can make learning easier, flatten the learning curve for several procedures, and facilitate various aspects of the surgical processes; (4) Conclusion: Technical complexity, equipment costs, and steep learning curves present significant obstacles to the widespread adoption of Mixed Reality and computer-assisted evaluation. The possibility of integration of distinct medical imaging modalities and innovative functionalities holds promise for clinical applications. However, it is important to consider issues related to usability, bias, and statistical analyses need consideration. Mixed Reality offers significant benefits, but there are still open challenges such as ergonomic issues, limited field of view, and battery autonomy that must be addressed to ensure widespread acceptance.

The design of a mixed-mode proportional-integral-derivative (PID) controller circuit using current-feedback operational amplifiers (CFOAs) as active components is proposed. With the same circuit topology, the proposed configuration of three CFOAs, four resistors, and two capacitors is capable of performing the PID controller in all four operation modes: namely voltage mode, trans-admittance mode, current mode, and trans-impedance mode. Numerous mathematical analyses are conducted to determine the controller performance under ideal and non-ideal conditions. Additionally, the mixed-mode second-order lowpass filter is suggested, and also used to examine the workability of the proposed mixed-mode PID controller in a feedback control structure. The proposed PID controller is implemented with the commercially available IC-type CFOA AD844, and the simulation results are presented to illustrate the functionality of the controller and its closed-loop control system.

When the majority of museums and other cultural institutions were shut down due to the pandemic, mixed museum visits became a hot issue. After the epidemic, mixed visits, in the opinion of many experts, would become the new norm for experiencing cultural content. Diverse types of merging online and onsite visits are already being investigated by researchers, not only to avoid the spread of a disease but also to enable visits of people who were previously excluded, such as persons in remote geographic areas or people with mobility challenges. The current work takes into account the Contextual Model of Museum Learning to define potential use scenarios for visits from people of different cultural backgrounds and offers an evaluation of current practices.