Safe Status Research Articles

Development of a starch-fermenting Zymomonas mobilis strain for bioethanol production

BackgroundBiorefinery using microorganisms to produce biofuels and value-added biochemicals derived from renewable biomass offers a promising alternative to meet our sustainable energy and environmental goals. The ethanologenic strain Zymomonas mobilis is considered as an excellent chassis for constructing microbial cell factories for diverse biochemicals due to its outstanding industrial characteristics in ethanol production, high specific productivity, and Generally Recognized as Safe (GRAS) status. Nonetheless, the restricted substrate range constrains its application.ResultsThe truncated ice nucleation protein InaK from Pseudomonas syringae was used as an autotransporter passenger, and α-amylase was fused to the C- terminal of InaK to equip the ethanol-producing bacterium with the capability to ferment renewable biomass. Western blot and flow cytometry analysis confirmed that the amylase was situated on the outer membrane. Whole-cell activity assays demonstrated that the amylase maintained its activity on the cell surface. The recombinant Z. mobilis facilitated the hydrolysis of starch into oligosaccharides and enabled the streamlining of simultaneous saccharification and fermentation (SSF) processes. In a 5% starch medium under SSF, recombinant strains containing Peno reached a maximum titer of 13.61 ± 0.12 g/L within 48 h. This represents an increase of 111.0% compared to the control strain's titer of titer of 6.45 ± 0.25 g/L.ConclusionsBy fusing the truncated ice nucleation protein InaK with α-amylase, we achieved efficient expression and surface display of the enzyme on Z. mobilis. This fusion protein exhibited remarkable enzymatic activity. Its presence enabled a cost-effective bioproduction process using starch as the sole carbon source, and it significantly reduced the required cycle time for SSF. This study not only provides an excellent Z. mobilis chassis for sustainable bioproduction from starch but also highlights the potential of Z. mobilis to function as an effective cellular factory for producing high-value products from renewable biomass.

Green bonds and clean energy stocks: Safe havens against global uncertainties? A wavelet quantile-based examination

The renewable energy sector has seen rapid investment growth over the last decade, with Green Bonds (GB) and Clean Energy Stocks (CLE) emerging as critical components of sustainable finance. Using daily data from 2013 to 2023, this study investigates the diverse roles of GB and CLE as potential safe havens under Geopolitical Risks (GEO) and Economic Policy Uncertainty (ECU). Applying an advanced Wavelet Quantile Regression (WQR) approach, we assess their effectiveness across different time horizons and risk conditions. The findings reveal that GB consistently offer stronger protection against economic risks, particularly in the short to mid-term, while CLE are more effective in shielding against geopolitical risks. Over longer periods, GB maintain their safe haven status, while CLE’s protective capabilities diminish. The results also highlight that GB and CLE behave differently across quantiles, with GB showing stability across extreme market conditions, making it a more reliable hedge overall. This study offers valuable policy insights for key stakeholders. Policymakers are encouraged to integrate GB and CLE into sustainable finance strategies, investors can mitigate risk by blending these assets, and portfolio managers should adjust allocations dynamically based on evolving market conditions. The results underscore the importance of green assets as hedges in times of global uncertainty, contributing to the growing literature on sustainable finance.

Safety assessment of enterocin-producing Enterococcus strains isolated from sheep and goat colostrum

BackgroundThis study investigates the safety evaluation of enterocin-producing 11 E. mundtii and two E. faecium strains previously isolated from small livestock colostrums. Enterococcus species do not possess Generally Recognized as Safe (GRAS) status. Hence, it is critical to scrutinize enterococci’s antibiotic resistance, virulence characteristics, and biogenic amine production capabilities in order to assess their safety before using them as starter or adjunct cultures.ResultsEnterococcus strains showed susceptibility to medically significant antibiotics. Multiple-drug resistance (MDR) was found in only E. faecium HC121.4, and its multiple antibiotic resistance (MAR) index was detected to be 0.22. The tetL and aph(3')-IIIa were the most commonly found antibiotic resistance genes in the strains. However, E. mundtii strains HC56.3, HC73.1, HC147.1, and E. faecium strain HC121.4 were detected to lack any of the antibiotic resistance genes examined in this study. Only E. mundtii HC166.3 showed hemolytic activity, while none of the strains engage in gelatinase activity. The strains were identified to have virulence factor genes with a low rate. None of the virulence factor genes could be detected in E. mundtii HC26.1, HC56.3, HC73.1, HC165.3, HC166.8, and E. faecium HC121.4. The E. mundtii HC73.2 strain displayed the highest presence of virulence factor genes, namely gelE, efaAfs, cpd, and ccf. Similarly, the E. mundtii HC112.1 strain showed a significant presence of genes efaAfm, ccf, and acm. There was no decarboxylation of histidine, ornithine, or lysine seen in any of the strains. Nevertheless, E. faecium HC121.4 and HC161.1 strains could decarboxylate tyrosine, but E. mundtii HC26.1, HC56.3, HC73.1, HC73.2, HC112.1, HC147.1, HC155.2, HC165.3, HC166.3, HC166.5, and HC166.8 strains only showed a limited capacity for tyrosine decarboxylation. None of the strains possessed the hdc, odc, or ldc genes, but all of them had the tdc gene.ConclusionThe E. mundtii HC56.3 and HC73.1 strains were deemed appropriate for utilization in food production. Using the remaining 11 strains as live cultures in food production activities could pose a possible risk to consumer health.

Epiphytic Yeasts from South Romania for Preventing Food Microbial Contamination.

Epiphytic yeasts represent an important source for the development of novel strategies aiming to combat food microbial contamination. The present study deals with the characterization of nine yeast strains belonging to Starmerella, Candida, Metschinikowia, Lachancea, Kodamaea and Pichia genera, isolated from the surface of plants from the Botanical Garden "Dimitrie Brandza" (Bucharest, Romania) for use as antimicrobial and probiotic agents. The tests involved the determination of the safe status, cell growth under stress conditions, and activity against pathogenic Candida and bacteria strains, respectively, as well as phytopathogenic filamentous fungi and lipolytic activity. None of the nine strains showed all the characteristics for virulence and pathogenicity, with the rare positive results being explained rather by their adaptability to the habitats of origin. The strains Lachancea thermotolerans CMGB-ST12 and Kodamaea ohmeri CMGB-ST19 grew at 37 °C; Metschnikowia reukaufii CMGB-ST21.2, M. reukaufii CMGB-ST.8.1 and M. reukaufii CMGB ST10 grew in the presence of 10% NaCl, while L. thermotolerans CMGB-ST12 and K. ohmeri CMGB-ST19 tolerated both acidic and alkaline pH values well (3.0 to 12.0). The studied yeast strains showed good antimicrobial activity against Candida krusei, Candida albicans and Gram-negative bacterial strains, with K. ohmeri CMGB-ST19 and Pichia membranaefaciens CMGB-ST53 inhibiting up to 100% the development of filamentous fungi. All the strains produced lipases for tributyrin hydrolysis, the best producer being Starmerella bombi CMGB-ST1, and only Candida magnoliae CMGB-ST8.2 tested positive against other probiotic yeasts. Overall, our nine yeast strains show high potential for industrial applications, for obtaining probiotic products and for preventing the development of a wide range of microbial food contaminants.

Optimization of Fermentative Parameters to Improve Gamma-Aminobutyric Acid (GABA) Production by Lactiplantibacillus plantarum B13

Gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the mammalian brain, and it possesses several physiological properties, such as depression reduction and anxiety release. GABA production by microbial synthesis is favoured over other methods like enzymatic and chemical synthesis due to the ease of operation and less formation of harmful pollutants. Lactic acid bacteria are widely applied for GABA production because of high GABA yield and their Generally Recognized as Safe (GRAS) status , which is critical in food and ingredient safety. In this study, various fermentation conditions, including incubation time, pH, temperature, monosodium glutamate (MSG) concentration, pyridoxal-5’-phosphate (PLP) concentration and glucose concentration were screened by one-factor-at-a-time strategy to achieve the optimal GABA production by a potential probiotic strain, Lactiplantibacillus plantarum B13. The result revealed the strain exhibited the optimal GABA production of 19.073 ± 0.5214 g/L with the highest GABA productivity of 0.424 g/L/h under fixed conditions: incubation time of 66 hours, pH 5.5, temperature of 35°C, MSG concentration of 5% (w/v), PLP concentration of 0.7 mM PLP and glucose concentration of 60 g/L. The findings of this study show that fermentation parameters are dependent on species and strains due to the different properties of glutamic acid decarboxylase enzymes and optimization of single parameters is important as a preliminary step to identify the range of fermentation factors that affect GABA yield prior further research endeavours. This study also has great implications for GABA production by L. plantarum B13 and provides a prerequisite for developing new healthy products enriched with GABA as daily supplements to support relaxation and regulate mood, reduce stress and promote better sleep.

TREAFET: Temperature-Aware Real-Time Task Scheduling for FinFET based Multicores

The recent shift in the VLSI industry from conventional MOSFET to FinFET for designing contemporary chip-multiprocessor (CMP) has noticeably improved hardware platforms’ computing capabilities, but at the cost of several thermal issues. Unlike the conventional MOSFET, FinFET devices experience a significant increase in circuit speed at a higher temperature, called temperature effect inversion (TEI), but higher temperature can also curtail the circuit lifetime due to self-heating effects (SHEs). These fundamental thermal properties of FinFET introduced a new challenge for scheduling time-critical tasks on FinFET-based multicores that how to exploit TEI towards improving performance while combating SHEs. In this work, TREAFET , a temperature-aware real-time scheduler, attempts to exploit the TEI feature of FinFET-based multicores in a time-critical computing paradigm. At first, the overall progress of individual tasks is monitored, tasks are allocated to the cores, and finally, a schedule is prepared. By considering the thermal profiles of the individual tasks and the current thermal status of the cores, hot tasks are assigned to the cold cores and vice-versa. Finally, the performance and temperature are balanced on-the-fly by incorporating a prudential voltage scaling towards exploiting TEI while guaranteeing the deadline and thermal safety. Moreover, TREAFET stimulates the average runtime frequency by employing an opportunistic energy-adaptive voltage spiking mechanism, in which energy saving during memory stalls at the cores is traded off during the time slice having the spiked voltage. Simulation results claim TREAFET maintains a safe and stable thermal status (peak temperature below 80 °C) and improves frequency up to 17% over the assigned value, which ensures legitimate time-critical performance for a variety of workloads while surpassing a state-of-the-art technique. The stimulated frequency in TREAFET also finishes the tasks early, thus providing opportunities to save energy by power gating the cores, and achieves a 24% energy delay product (EDP) gain on average.

Developing Starter Culture from Lactic Acid Bacteria Isolated from Cow Milk for the Production of Nigerian “Nunu”

Starter cultures are mostly organisms deliberately introduced into food as a single culture or mixture of cultures to bring about fermentation and production of desired characteristics in food through metabolic and enzymatic activities. Organisms in this category have attained the generally regarded as safe (GRAS) status, are majorly lactic acid bacteria with food acidification and production of natural preservatives potential. This research was aimed at developing a starter culture for the production of local dairy drink “Nunu” Cow milk was obtained from “Gaa Mobolohunduro”, Tanke, Ilorin, Kwara State, Nigeria. Potential lactose fermenters were isolated on MRS agar in pour plate technique. The isolates were characterized and identified using standard procedures. Lactic Acid Bacteria isolated were inoculated as a starter culture for milk fermentation and the products were subjected to pH, titratable acid and organoleptic assessments following standard procedures. Two of the isolated lactic acid bacteria with the best fermentation products were identified through molecular technique. A total of six isolates were obtained and identified as Lactococcus sp., Leuconostoc sp., Pediococcus sp. and Lactobacillus spp. The pH ranges from 5.46±0.01 to 6.03±0.01while the titratable acidity ranged from 0.244±0.004 to 0.526±0.006. The moisture content ranged from 90.50±0.30 to 91.75±0.35. The fermentation product from the isolated Lactobacillus sp. scored 80 % during the organoleptic assessment. Molecular evidence confirmed the two isolates as Lactic Acid Bacteria. In conclusion, isolated species could be used as potential organisms for milk fermentation.

Effects of Genotype and Growing Year on the Nutritional Composition and Pasting Properties of Glabrous Canary Seed (Phalaris canariensis L.) Groat Flours.

Canary seed flour is a new food ingredient that the United States Food and Drug Administration (FDA) and Health Canada recently granted Generally Regarded as Safe (GRAS) status. Stability in nutritional composition and functional properties is an essential characteristic of food ingredients for consistency in nutritional quality and performance in processing. This work assessed the effect of genotypic and environmental variation on the nutritional (protein, starch, amylose, oil, dietary fiber, minerals and fat-soluble vitamins) and pasting (as measured in viscosity (peak, trough, breakdown, final, and setback), peak time, and pasting temperatures) properties of Canary seed. The samples included four Canary seed varieties grown in randomized complete block design experiments at one location for two growing seasons. In general, the nutritional composition of Canary seed flour was not affected by genotype, growing year, and their interaction except for starch content, which was significantly affected by the growing year (p < 0.0001), and iron content, which was affected by genotypic variation (p < 0.0001). The pasting properties of Canary seed flour were significantly (p < 0.001) affected by both genotypic and growing year variation but not their interaction. Our results suggest that the food industry should measure starch and iron content prior to processing to ensure consistency in nutritional labeling. Also, for those applications where starch pasting properties are essential, the manufacturer should consider measuring the RVA pasting viscosities for every batch of raw material. The results have provided the baseline knowledge of which nutritional or functional properties of Canary seed flour can be improved through breeding and agronomy programs to ensure the reliability of Canary seed as an ingredient.

Saccharomyces cerevisiae oral immunization in mice using multi-antigen of the African swine fever virus elicits a robust immune response.

African swine fever virus (ASFV) is one of the most complex viruses. ASFV is a serious threat to the global swine industry because no commercial vaccines against this virus are currently available except in Vietnam. Moreover, ASFV is highly stable in the environment and can survive in water, feed, and aerosols for a long time. ASFV is transmitted through the digestive and respiratory tract. Mucosal immunity is the first line of defense against ASFV. Saccharomyces cerevisiae (SC), which has been certified by the U.S. Food and Drug Administration and has a generally recognized as safe status in the food industry, was used for oral immunization in this study. ASFV antigens were effectively expressed in recombinant SC strains with high DNA copy numbers and stable growth though surface display technology and chromosome engineering (δ-integration). The recombinant SC strains containing eight ASFV antigens-KP177R, E183L, E199L, CP204L, E248R, EP402R, B602L, and B646L- induced strong humoral and mucosal immune responses in mice. There was no antigenic competition, and these antigens induced Th1 and Th2 cellular immune responses. Therefore, the oral immunization strategy using recombinant SC strains containing multiple ASFV antigens demonstrate potential for future testing in swine, including challenge studies to evaluate its efficacy as a vaccine against ASFV.

Time-variant safe haven currencies

This study investigates the temporal variation in the safe haven status of the Japanese yen, Swiss franc, and U.S. dollar, using a rolling GARCH-in-mean model. Until the global financial crisis, the U.S. dollar was likely a safe haven currency, while the Swiss franc has been one since the crisis. The Japanese yen later emerged as a safe haven and retained its status during the U.S.–China trade friction and the COVID-19 pandemic. The findings suggest that national economic policies, market liquidity, and socio-political events affect safe haven status.

Financial Distress Assessment Through Altman Z-Score

This research aims to assess the financial distress condition of PT Waskita Karya Tbk (WSKT), a state-owned construction company in Indonesia, from 2017 to 2022 using the Altman Z-Score model. The Altman Z-Score combines financial ratios to predict the likelihood of bankruptcy. Secondary data derived from WSKT’s financial statements were analyzed quantitatively using the Z-Score formula for non-manufacturing firms. The findings indicate that WSKT has experienced significant financial distress. In 2017 and 2022, WSKT was in the Distress Zone with scores of 0.96570 and 0.78271 respectively. The company was also in distress in 2020 with a negative score of -1.12303. For 2018, 2019 and 2021, WSKT was in the Grey Zone with scores of 1.51502, 1.05679 and 1.17451. WSKT did not achieve Safe Zone status during the six years examined. Overall, 66.67% of the period reviewed falls in the Distress Zone, predicting a high bankruptcy risk for WSKT. To address this financial vulnerability, recommendations include conducting operational optimization to enhance efficiency and profitability, diversifying business portfolios to tap into promising segments, and proactively monitoring financial health using analytical models like Altman Z-Score. With disciplined implementation of strategies to rectify distressed ratios, WSKT can achieve financial stability.

Prediction Model for Safe Operation of Pumping Stations Optimized by the Sparrow Search Algorithm and BP Neural Network

The pumping station is one of the critical parts of the hydraulic structure in China. Traditional forecasting methods are limited in accuracy, time-consuming, and high cost, resulting in limited data availability. Therefore, simulation model analysis based on soft computation is a realistic and valuable alternative. This article intends to use the BP neural network to predict the safe operation status of pump stations and optimize the initial threshold and weight information of the BP network using the sparrow search algorithm (SSA) to improve the accuracy and generalization ability of the model. In addition, to more accurately reflect the correlation between various influencing factors and the safe operation status of the pumping station, the entropy weight method and the analytic hierarchy process were used to obtain the comprehensive weights of each main influencing factor. The experimental results show that the SSA-BP model can accurately predict the safe operation status of pumping stations, and compared with other traditional models, the SSA-BP model has better convergence and higher accuracy. This model provides a new approach for predicting the safe operation of pumping stations and has particular reference significance for predicting the safe operation of other pumping stations.

Engineering the Tat-secretion pathway of Bacillus licheniformis for the secretion of cytoplasmic enzyme arginase.

The industrial bacterium Bacillus licheniformis has long been used as a microbial factory for the production of enzymes due to its ability to secrete copious amounts of native extracellular proteins and its generally regarded as safe (GRAS) status. However, most attempts to use B. licheniformis to produce heterologous and cytoplasmic enzymes primarily via the general secretory (Sec) pathway have had limited success. The twin-arginine transport (Tat) pathway offers a promising alternative for the extracellular export of Sec-incompatible proteins because it transports full, correctly folded proteins. However, compared to the Sec pathway, the yields of the Tat pathway have historically been too low for commercial use. To improve the export efficiency of the Tat pathway, we identified the optimal Tat-dependent signal peptides and increased the abundance of the Tat translocases, the signal peptidase (SPase), and the intracellular chaperones. These strategic modifications significantly improved the Tat-dependent secretion of the cytoplasmic enzyme arginase into the culture medium using B. licheniformis. The extracellular enzymatic activity of arginase showed a 5.2-fold increase after these modifications. Moreover, compared to the start strain B. licheniformis 0F3, the production of extracellular GFP was improved by 3.8 times using the strategic modified strain B. licheniformis 0F13, and the extracellular enzymatic activity of SOX had a 1.3-fold increase using the strain B. licheniformis 0F14. This Tat-based production chassis has the potential for enhanced production of Sec-incompatible enzymes, therefore expanding the capability of B. licheniformis as an efficient cellular factory for the production of high-value proteins. KEY POINTS: • Systematic genetic modification of Tat-pathway in B. licheniformis. • Significant enhancement of the secretion capacity of Tat pathway for delivery the cytoplasmic enzyme arginase. • A new platform for efficient extracellular production of Sec-incompatible enzymes.

Characterization of Lactobacillus spp. as Probiotic and Antidiabetic Potential Isolated from Boza, Traditional Fermented Beverage in Turkey.

Boza, a cereal-based beverage popular in southeast Europe, is fortified with probiotics and is believed to positively impact the composition of the gut microflora. This investigation focused on fermented cereal-based beverage boza to identify strains of probiotic Lactobacillus spp. capable of inhibiting carbohydrate-hydrolysing enzymes α-glucosidase (AG) and α-amylase (AA). The isolated bacterial strains underwent a comprehensive assessment, including biochemical, molecular, and probiotic trait analyses such as tolerance survivability, adhesion, safety, and health-promoting attributes. We evaluated the inhibitory potential of the supernatant, cell lysate, and intact cells of Lactobacillus spp. Molecular analysis has revealed that isolates RAMULAB30 and RAMULAB29 exhibit a significant genetic similarity (>97%) to Lacticaseibacillus paracasei and Limosilactobacillus fermentum, respectively. These findings are documented in the NCBI database. They exhibited significant resistance to gastrointestinal and intestinal fluids, also indicating their potential for adhesion. Additionally, the isolates showed a significant antibacterial activity, particularly against Micrococcus luteus. They showed resistance to vancomycin and methicillin antibiotics but were more susceptible to streptomycin and ampicillin. Furthermore, the strains demonstrated antioxidant properties. To ensure their safety, a haemolytic assay was conducted despite their general recognition as safe (GRAS) status. The study primarily aimed to evaluate the inhibitory effects of the extract on enzymes AG and AA. Bacterial isolates demonstrated a significant inhibitory activity against both enzyme AG (32%-67% inhibition) and enzyme AA (18%-46% inhibition) in different forms, including supernatant (CS), lysed extract (CE), and intact cell (IC). These findings underscore the potential of bacterial isolates to inhibit the enzyme activity effectively. Furthermore, the L. fermentum RAMULAB29 and L. paracasei RAMULAB30 strains exhibit remarkable antidiabetic potential. Food products incorporating these strains have promising prospects as nutraceuticals, providing improved health benefits.

Social Potential of Teachers in the Professional Environment

The article presents the results of a study on the social potential of teachers in a professional environment. Social potential was understood as intangible resources of commitment, trust, norms of reciprocity and networks of personal connections among teachers that enable cooperation and exchange of knowledge. The research concept was based on the theory of social capital, which has been popular for some time in economics and politics, but also in the social sciences. It is clarified that in the education system in the current conditions of formation of a new information society, the role and importance of the teacher’s social potential is increasing. Therefore, the work of a modern teacher takes place in the context of the expectations that society places on him or her. The study distinguished four main variables: participation and cooperation, interpersonal relationships and communication, trust and credibility, reciprocity and loyalty. The results reveal the division of the respondents into two groups. The first of them are teachers who, in their assessments, create a positive image of their own social potential, especially in the case of participation, involvement and cooperation. The second group are teachers who are more skeptical and ambivalent. This is especially visible in the dimensions of reciprocity and loyalty as well as relationships and communication. Teachers do not strive to evaluate their own environment negatively. Perhaps they do not notice or do not want to notice uncomfortable, difficult or conflicting issues. It can therefore be assumed that this is a confirmation of the façade and superficiality of behavior and relationships among teachers, and the desire to maintain a safe status quo.

Evaluation of the dynamic additional impact about foundation pit construction on the existing adjacent subway station with the PBA method

In this paper, relying on the Tianhe East Station project of Guangzhou Subway Line 11, the deformation and mechanical evolution rules about the existing adjacent stations with the pile-beam-arch (PBA) method in the all excavation stages about the new foundation pit is evaluated through numerical calculation method. It was revealed that that the largest displacement about the existing station’s side wall and the middle column reaches 2.1mm and 1.5mm respectively during the excavation about the foundation pit. Due to the impact about construction of the foundation pit, the uplifting phenomenon occurs on the side of the foundation pit, and the maximum uplifting value reaches 0.4mm. When the construction about foundation pit is finished, the whole station floor shows a deformation form of “left lower torsion”, and the largest compressive stresses as well as tensile stresses about the side wall and the bottom plate do not reach the standard limiting value, and the whole is being a relative safe status. With the increase about the excavation depths, the axial forces about the middle column gradually increase to 7016kN, and the incremental axial forces about the middle column after the construction is completed accounts for about 11.7% of the axial force of initial construction. Therefore, the disturbance effect about foundation pit construction on the adjacent station with the PBA method can not be ignored. The research result in this paper may offer some important references for the construction and design of similar cases.

Early warning system for load distribution in automated warehouse based on seismic resilience evaluation

As an integral part of the storage system in an automated warehouse, the racking structure can be altered for seismic performance by adjusting the distribution of loads, which is different from other building structures with uncertain load distribution. Therefore, there are two ways to improve the seismic resilience of racks: improving structural response and optimizing load distributions. The former has been widely studied, while the latter is rarely noticed. The objective of this study is to develop a method of seismic resilience evaluation and an early warning system of over-limit load distribution for racks with spine bracing. Based on the influence of load distribution on the seismic performance of racks with spine bracing, the contribution value of load positions was calculated to make additional corrections to the maximum stress ratio in load distribution samples obtained by genetic algorithm. Then, a formula for seismic resilience evaluation for racks with spine bracing was derived and its accuracy was validated to quantify the safety level of different load distribution patterns. An early warning system for over-limit load distribution was established and codified in a software package. A traffic light system including Green, Red, and Yellow were employed in the software package as an indicator of the load distribution safety level of racks. Finally, the system developed was applied to the actual racks in an automated tobacco warehouse in Nanjing. By adjusting loads in the least number of steps and the shortest distance, the racks changed from dangerous status (red light) to safe status (green light). The early warning system allows for rapid evaluation for the seismic resilience of racks without real-time finite element analysis, and is proved to be applicable, efficient and reliable.

Improved production of lactiplantibacillus plantarum RO30 exopolysaccharide (REPS) by optimization of process parameters through statistical experimental designs

BackgroundIn investigating of (exopolysaccharide) EPS from unconventional sources, lactic acid bacteria have a vital role due to their generally recognized as safe (GRAS) status. EPSs have diverse applications such as drug delivery, antimicrobial activity, surgical implants, and many more in many sectors. Despite being important, the main hindrance to the commercial application of these significant biopolymers is low productivity. Therefore, this study primarily focuses on optimizing physio-chemical conditions to maximize the previously produced EPS from probiotic Lactiplantibacillus plantarum RO30 (L. plantarum RO30) using one factor at a time (OFAT) and method Response Surface Methodology (RSM).ResultsThe EPS obtained from L. plantarum RO30 named REPS. The medium formulation for REPS production using the OFAT method revealed that sucrose (20 g/L, beef extract (25 g/L), and ammonium sulfate at 4 g/L concentration were the optimum carbon, organic and inorganic nitrogen sources, and REPS yield was increased up to 9.11 ± 0.51 g/L. RSM experiments revealed that, a greatly significant quadratic polynomial attained from the Central Composite Design (CCD) model was fruitful for specifying the most favorable cultural conditions that have significant consequences on REPS yield. The maximal amount of REPS (10.32 g/L) was formed by: sucrose (40 g/L), beef extract (25 g/L), pH (5.5), incubation temperature (30 °C), and incubation period (72 h). A high closeness was obtained between the predicted and experimental values and it displayed the efficiency of the RSM.ConclusionThis study was conducted to reinforce REPS production in the probiotic LAB L. plantarum RO30 by utilizing various experimental parameters. The maximum REPS yield of 10.32 g/L was attained under the circumstances optimized in the study.

Enzymatic and antibacterial activity of the recombinant endolysin PVP-SE1gp146 expressed in Hansenula polymorpha

Antibiotic resistance, a major global concern, highlights the need for discovering alternative therapies. Recently, endolysins have garnered attention as antibacterial tools with a lower resistance development rate compared to conventional antibiotics, and their production in various expression hosts holds significance. Given its generally recognized as safe (GRAS) status and other advantages, Hansenula polymorpha offers a promising host for endolysin production. PVP-SE1gp146 originates from the Salmonella Enteritidis-specific phage PVP-SE1, which has been previously characterized. We inserted the PVP-SE1gp146 coding gene into the H. polymorpha expression vector pHIPX4. The resulting recombinant, pHIPX4-PVP-SE1gp146, was then introduced into H. polymorpha NCYC495 to facilitate the production of the endolysin PVP-SE1gp146. The expression level of the PVP-SE1gp146 protein was assessed, and it was determined to be approximately 43 mg/l of yeast culture medium. The enzymatic (muralytic) activity of this endolysin was also evaluated, corresponding to the version produced by the E. coli Bl21 strain. The endolysin exhibited admissible antibacterial activity against several gram-negative species, including P. aeruginosa, E. coli, and A. baumannii, while showing an almost negligible impact on K. pneumoniae. Endolysin production within GRAS-approved hosts holds potential for combating antibiotic-resistant bacteria. Challenges involve optimizing concentrations, targeting gram-negative species and improving attachment to bacterial cell walls. Addressing these issues requires dedicated research in endolysin engineering and a comprehensive evaluation of their production in diverse expression hosts.

200 The Effects of Whole Hemp Seed Supplementation on Health Status, Voluntary Physical Activity, Fecal Quality, and Fermentative End-Products in Healthy Adult Dogs

Abstract The cultivation and utilization of hemp and hemp-derived products have increased at a rapid pace since the enactment of the 2018 farm bill which provided changes in regulation related to cannabis. Although there are three hemp-derived ingredients that have been generally recognized as safe status for human consumption by the FDA, no ingredients have been approved by the FDA for consumption by companion animals or defined by AAFCO. Hemp seeds have the potential to act as a rich source of plant protein, polyunsaturated fatty-acids, and dietary fiber in companion animal nutrition. The objective of this study was to evaluate the effects of supplementing ground whole hemp seed in a baked chew on voluntary physical activity, fecal characteristics, fermentative-end products, and overall health status in healthy adult dogs. All animal procedures were approved by the University of Illinois Institutional Animal Care and Use Committee and were in accordance with the United States Public Health Service Policy on Humane Care and Use of Laboratory Animals. Thirty adult beagle dogs were used in a complete randomized design, with ten dogs assigned to each treatment. The dogs were fed the same commercially available, complete and balanced diet and were supplemented with one of three baked chews containing either a high inclusion of ground hemp seed (50%), a low inclusion of ground hemp seed (25%), or cellulose. The baked chews were fed at a level of 10% of the daily intake for 28 d and were formulated to be similar in macronutrient composition, approximately 25% fat, 20% protein, and 25% total dietary fiber. Duration and intensity of voluntary physical activity was measured using activity monitors for 4 d before supplementation and at the end of the supplementation period (d 24 to 27). Fasted blood and a fresh fecal sample were taken on d 0, 14, and 28 to monitor health status, blood variables, fecal characteristics and fermentative-end product concentrations. The baked chews were well accepted by the dogs with minimal intake refusals, and all dogs remained clinically healthy for the duration of the experimental period. No differences in fecal score or fecal pH were observed among the treatment groups (P &amp;gt; 0.05). The treatment did not have an effect on fecal concentrations of volatile fatty acids including acetate, propionate, butyrate, valerate, isovalerate, and isobutyrate (P &amp;gt; 0.05). Concentrations of fecal ammonia, phenols, and indoles were also similar among the groups (P &amp;gt; 0.05). No adverse effects were observed, and the data suggest that in a baked chew the whole ground hemp seed was comparable to the cellulose control.