Mitochondria-associated membrane collapse impairs TBK1-mediated proteostatic stress response in ALS.

The organelle contact site of the endoplasmic reticulum and mitochondria, known as the mitochondria-associated membrane (MAM), is a multifunctional microdomain in cellular homeostasis. We previously reported that MAM disruption is a common pathological feature in amyotrophic lateral sclerosis (ALS); however, the precise role of MAM in ALS was uncovered. Here, we show that the MAM is essential for TANK-binding kinase 1 (TBK1) activation under proteostatic stress conditions. A MAM-specific E3 ubiquitin ligase, autocrine motility factor receptor, ubiquitinated nascent proteins to activate TBK1 at the MAM, which results in ribosomal protein degradation. MAM or TBK1 deficiency under proteostatic stress conditions resulted in increased cellular vulnerability in vitro and motor impairment in vivo. Thus, MAM disruption exacerbates proteostatic stress via TBK1 inactivation in ALS. Our study has revealed a proteostatic mechanism mediated by the MAM-TBK1 axis, highlighting the physiological importance of the organelle contact sites.

Phosphorylation of aryl hydrocarbon receptor interacting protein by TBK1 negatively regulates IRF7 and the type I interferon response.

The innate antiviral response to RNA viruses is initiated by sensing of viral RNAs by RIG-I-like receptors and elicits type I interferon (IFN) production, which stimulates the expression of IFN-stimulated genes that orchestrate the antiviral response to prevent systemic infection. Negative regulation of type I IFN and its master regulator, transcription factor IRF7, is essential to maintain immune homeostasis. We previously demonstrated that AIP (aryl hydrocarbon receptor interacting protein) functions as a negative regulator of the innate antiviral immune response by binding to and sequestering IRF7 in the cytoplasm, thereby preventing IRF7 transcriptional activation and type I IFN production. However, it remains unknown how AIP inhibition of IRF7 is regulated. We show here that the kinase TBK1 phosphorylates AIP and Thr40 serves as the primary target for TBK1 phosphorylation. AIP Thr40 plays critical roles in regulating AIP stability and mediating its interaction with IRF7. The AIP phosphomimetic T40E exhibited increased proteasomal degradation and enhanced interaction with IRF7 compared with wildtype AIP. AIP T40E also blocked IRF7 nuclear translocation, which resulted in reduced type I IFN production and increased viral replication. In sharp contrast, AIP phosphonull mutant T40A had impaired IRF7 binding, and stable expression of AIP T40A in AIP-deficient mouse embryonic fibroblasts elicited a heightened type I IFN response and diminished RNA virus replication. Taken together, these results demonstrate that TBK1-mediated phosphorylation of AIP at Thr40 functions as a molecular switch that enables AIP to interact with and inhibit IRF7, thus preventing overactivation of type I IFN genes by IRF7.

Burkholderia pseudomallei Bacteria in Ornamental Fish Tanks, Vientiane, Laos, 2023.

In 2019, a melioidosis case in Maryland, USA, was shown to have been acquired from an ornamental fish tank contaminated with Burkholderia pseudomallei bacteria, likely derived from Southeast Asia. We investigated the presence of B. pseudomallei in ornamental fish tanks in the endemic area of Vientiane, Laos.

Xiashengella succiniciproducens gen. nov., sp. nov., a succinate-producing bacterium isolated from an anaerobic digestion tank in the family Marinilabiliaceae of the order Bacteroidales.

A strictly anaerobic, motile bacterium, designated as strain Ai-910T, was isolated from the sludge of an anaerobic digestion tank in China. Cells were Gram-stain-negative rods. Optimal growth was observed at 38 °C (growth range 25-42 °C), pH 8.5 (growth range 5.5-10.5), and under a NaCl concentration of 0.06% (w/v) (range 0-2.0%). Major cellular fatty acids were iso-C15 : 0 and anteiso-C15 : 0. The respiratory quinone was MK-7. Using xylose as the growth substrate, succinate was produced as the fermentation product. Phylogenetic analysis based on the 16 S rRNA gene sequences indicated that strain Ai-910T formed a distinct phylogenetic lineage that reflects a new genus in the family Marinilabiliaceae, sharing high similarities to Alkaliflexus imshenetskii Z-7010T (92.78%), Alkalitalea saponilacus SC/BZ-SP2T (92.51%), and Geofilum rubicundum JAM-BA0501T (92.36%). Genomic similarity (average nucleotide identity and digital DNA-DNA hybridization) values between strain Ai-910T and its phylogenetic neighbors were below 65.27 and 16.90%, respectively, indicating that strain Ai-910T represented a novel species. The average amino acid identity between strain Ai-910T and other related members of the family Marinilabiliaceae were below 69.41%, supporting that strain Ai-910T was a member of a new genus within the family Marinilabiliaceae. Phylogenetic, genomic, and phenotypic analysis revealed that strain Ai-910T was distinguished from other phylogenetic relatives within the family Marinilabiliaceae. The genome size was 3.10 Mbp, and the DNA G + C content of the isolate was 42.8 mol%. Collectively, differences of the phenotypic and phylogenetic features of strain Ai-910T from its close relatives suggest that strain Ai-910T represented a novel species in a new genus of the family Marinilabiliaceae, for which the name Xiashengella succiniciproducens gen. nov., sp. nov. was proposed. The type strain of Xiashengella succiniciproducens is Ai-910T (= CGMCC 1.17893T = KCTC 25,304T).

New insights into the role of the root system of epiphytic bromeliads: comparison of root and leaf trichome functions in acquisition of water and nutrients.

BACKGROUND: In epiphytic bromeliads, the roots used to be considered poorly functional organs in the processes of absorption and metabolization of water and nutrients, while the leaves always acted as protagonists in both functions. More recent discoveries have been changing this old view of the root system. SCOPE: In this review, we will address the old thoughts of the scientific community regarding the function performed by the roots of epiphytic bromeliads (mere holdfast structures with low physiological activity) and the importance of a reduced or lack of root system for the emergence of epiphytism. We will present indirect and direct evidence that contradicts this older hypothesis. Furthermore, the importance of the root absorptive function mainly for juvenile tankless epiphytic bromeliads and the characteristics of the root absorption process of adult epiphytic tank bromeliads will be thoroughly discussed in physiological aspects. Finally, some factors (species, substrate, environmental conditions) that influence the absorptive capability of the roots of epiphytic tank bromeliads will also be considered in this review, highlighting the importance that the absorptive role of the roots have for the plasticity of bromeliads that live on trees, which is an environment characterized by the intermittent availability of water and nutrients. CONCLUSIONS: The roots of tank-forming epiphytic bromeliads play important roles in the absorption and metabolization of nutrients and water. The importance of roots stands out mainly for juvenile tankless bromeliads since the root is the main absorptive organ. In larger plants with tank, although the leaves become the protagonists in the resource acquisition process, the roots complement the absorptive function of the leaf trichomes, resulting in a better growth of the bromeliad. The physiological and biochemical properties of the processes of absorption and distribution of resources in the tissues seem to differ between absorption by trichomes and roots.

Methylomonas defluvii sp. nov., a type I methane-oxidizing bacterium from a secondary sedimentation tank of a wastewater treatment plant.

An aerobic methanotroph was isolated from a secondary sedimentation tank of a wastewater treatment plant and designated strain OY6T. Cells of OY6T were Gram-stain-negative, pink-pigmented, motile rods and contained an intracytoplasmic membrane structure typical of type I methanotrophs. OY6T could grow at a pH range of 4.5-7.5 (optimum pH 6.5) and at temperatures ranging from 20 °C to 37 °C (optimum 30 °C). The major cellular fatty acids were C14 : 0, C16 : 1ω7c/C16 : 1ω6c and C16 : 1ω5c; the predominant respiratory quinone was MQ-8. The genome size was 5.41 Mbp with a DNA G+C content of 51.7 mol%. OY6T represents a member of the family Methylococcaceae of the class Gammaproteobacteria and displayed 95.74-99.64 % 16S rRNA gene sequence similarity to the type strains of species of the genus Methylomonas. Whole-genome comparisons based on average nucleotide identity (ANI) and digital DNA-DNA hybridisation (dDDH) confirmed that OY6T should be classified as representing a novel species. The most closely related type strain was Methylomonas fluvii EbBT, with 16S rRNA gene sequence similarity, ANI by blast (ANIb), ANI by MUMmer (ANIm) and dDDH values of 99.64, 90.46, 91.92 and 44.5 %, respectively. OY6T possessed genes encoding both the particulate methane monooxygenase enzyme and the soluble methane monooxygenase enzyme. It grew only on methane or methanol as carbon sources. On the basis of phenotypic, genetic and phylogenetic data, strain OY6T represents a novel species within the genus Methylomonas for which the name Methylomonas defluvii sp. nov. is proposed, with strain OY6T (=GDMCC 1.4114T=KCTC 8159T=LMG 33371T) as the type strain.

Understanding exopolysaccharide byproduct formation in Komagataella phaffii fermentation processes for recombinant protein production.

BACKGROUND: Komagataella phaffii (Pichia pastoris) has emerged as a common and robust biotechnological platform organism, to produce recombinant proteins and other bioproducts of commercial interest. Key advantage of K. phaffii is the secretion of recombinant proteins, coupled with a low host protein secretion. This facilitates downstream processing, resulting in high purity of the target protein. However, a significant but often overlooked aspect is the presence of an unknown polysaccharide impurity in the supernatant. Surprisingly, this impurity has received limited attention in the literature, and its presence and quantification are rarely addressed. RESULTS: This study aims to quantify this exopolysaccharide in high cell density recombinant protein production processes and identify its origin. In stirred tank fed-batch fermentations with a maximal cell dry weight of 155 g/L, the polysaccharide concentration in the supernatant can reach up to 8.7 g/L. This level is similar to the achievable target protein concentration. Importantly, the results demonstrate that exopolysaccharide production is independent of the substrate and the protein production process itself. Instead, it is directly correlated with biomass formation and proportional to cell dry weight. Cell lysis can confidently be ruled out as the source of this exopolysaccharide in the culture medium. Furthermore, the polysaccharide secretion can be linked to a mutation in the HOC1 gene, featured by all derivatives of strain NRRL Y-11430, leading to a characteristic thinner cell wall. CONCLUSIONS: This research sheds light on a previously disregarded aspect of K. phaffii fermentations, emphasizing the importance of monitoring and addressing the exopolysaccharide impurity in biotechnological applications, independent of the recombinant protein produced.

Development of an HSV-1 production process involving serum-reduced culturing and bead-to-bead transfer.

Herpes simplex virus type 1 (HSV-1) plays an important role in the field of gene therapy and viral vaccines, especially as an oncolytic virus. However, the mass production of HSV-1 viral vectors remains a challenge in the industry. In this study, a microcarrier-mediated serum-reduced medium culture was used to improve the bioprocess of HSV-1 production and increase HSV-1 yields. The composition of the culture media, which included a basal medium, serum concentration, and glutamine additive, was optimized. The process was successfully conducted in a 1 L bioreactor, and virus production was threefold greater than that of conventional processes with a 10% serum medium. The bead-to-bead transfer process was also developed to further increase scalability. In spinner flasks, the detachment rate increased from 49.4 to 80.6% when combined agitation was performed during digestion; the overall recovery proportion increased from 37.9 to 71.1% after the operational steps were optimized. Specifically, microcarrier loss was reduced during aspiration and transfer, and microcarriers and detached cells were separated with filters. Comparable cell growth was achieved with the baseline process using 2D culture as the inoculum by exchanging the subculture medium. To increase virus production after bead-to-bead transfer, critical parameters, including shear stress during digestion, TrypLE and EDTA concentrations in the subculture, and the CCI, were identified from 47 parameters via correlation analysis and principal component analysis. The optimized bead-to-bead transfer process achieved an average of 90.4% overall recovery and comparable virus production compared to that of the baseline process. This study is the first to report the optimization of HSV-1 production in Vero cells cultured on microcarriers in serum-reduced medium after bead-to-bead transfer. KEY POINTS: • An HSV-1 production process was developed that involves culturing in serum-reduced medium, and this process achieved threefold greater virus production than that of traditional processes. • An indirect bead-to-bead transfer process was developed with over 90% recovery yield in bioreactors. • HSV-1 production after bead-to-bead transfer was optimized and was comparable to that achieved with 2D culture as inoculum.

Methemoglobinemia screening and treatment in tank warfare survivors.

INTRODUCTION: Methemoglobinemia, characterized by the conversion of functional hemoglobin to methemoglobin, can significantly impede tissue oxygenation. Prompt diagnosis and treatment of methemoglobinemia are critical to optimizing clinical outcomes. Although the underlying etiology of methemoglobinemia is often attributed to a medication reaction or chemical exposure, its association with battlefield trauma remains underexplored. This case series explores the presence of methemoglobinemia in nine soldiers evacuated from tanks targeted by explosives, shedding new light on screening needs and treatment strategies. CASES DESCRIPTION: Nine combat trauma patients with methemoglobinemia were admitted to Soroka Medical Center over a two-month period. Detailed case descriptions illustrate the diverse presentations and treatment responses. Notably, the administration of methylene blue resulted in rapid methemoglobin reductions and an improvement in oxygenation without any observed side effects. DISCUSSION: This series highlights an unexpected consequence of an explosion within an armored fighting vehicle and the challenges related to standard pulse oximetry interpretation and accuracy in the presence of methemoglobinemia, emphasizing the need for vigilant monitoring and co-oximetry utilization. Additionally, the coexistence of carboxyhemoglobin further warrants attention due to its synergistic and deleterious effects on oxygen delivery. Collaborative efforts with military authorities should aim to explore the underlying mechanisms associated with trauma and methemoglobinemia and optimize battlefield care. CONCLUSION: This case series underscores the significance of methemoglobinemia screening in combat trauma patients, and advocates for systematic co-oximetry utilization and methylene blue availability in combat zones. Early detection and intervention of methemoglobinemia in combat soldiers are often difficult in the context of battlefield injuries but are necessary to mitigate the potentially fatal consequences of this condition.

Ocular injury from saltwater coral palytoxin.

We describe a case of a young male who presented to the emergency department with unilateral eye pain, blurred vision, conjunctival injection, and ocular pH of 9, one day after direct ocular exposure to palytoxin (PTX) from coral in a home saltwater fish tank. Although uncommon, ocular PTX toxicity is a potentially vision-threatening condition that requires prompt recognition. This case report documents the successful management of presumed ocular PTX exposure and suggests additional workup and treatment considerations for future patients.

Microbial communities in feed, bedding material, and bulk milk - experiences from a feeding trial.

There is an increasing interest in the microbiota of the dairy value chain, from field to fork. Studies to understand the effects of environmental, feed and management factors on the raw milk microbiota have been performed to elucidate the origin of the bacteria and find ways to control the presence or absence of specific bacteria. In this study, we explored the microbiota in feedstuff, bedding material and milk on a Swedish dairy farm to investigate the effects of feeding different silages on the bacterial compositions throughout the dairy value chain. Three ensiling treatments were evaluated: without additive, with acid treatment, and with inoculation of starter culture. The silage treatments were fed as partial mixed rations to 67 dairy cows for 3 weeks each, with one treatment fed twice to evaluate if a potential change in milk microbiota could be repeated. The highest average total bacteria counts were found in the used bedding material (9.6 log10 cfu/g), while milk showed the lowest (3.5 log10 cfu/g). Principal coordinate analysis of the weighted UniFrac distance matrix showed clear separation between 3 clusters of materials: 1) herbage, 2) silage and partial mixed ration, and 3) used bedding material and milk. Surprisingly, the expected effect of the ensiling treatments on silage microbiota was not clear. Transfer of major bacteria from the silages and resulting partial mixed rations to the used bedding material was observed, but rarely to milk. The milk microbiota showed most resemblance to that of the used bedding material. Lactobacillus was a major genus in both feed and milk, but investigations at amplicon sequence variant level showed that in most cases the sequences differed between materials. However, low total bacteria count in the milk in combination with a high diversity suggests that results may be biased due to environmental contamination of the milk samples. Considering that the study was performed on a research farm, strict hygienic measures during the feeding experiment may have contributed to the low transfer of bacteria from feed to milk.

The microbiota of ensiled forages and of bulk tank milk on dairy cattle farms in northern Sweden - a case study.

Factors contributing to variations in the quality and microbiota of ensiled forages and in bulk tank microbiota in milk from cows fed different forages were investigated. Nutritional quality, fermentation parameters and hygiene quality of forage samples and corresponding bulk tank milk samples collected in 3 periods from 18 commercial farms located in northern Sweden were compared. Principal coordinates analysis revealed that the microbiota in forage and bulk milk, analyzed using 16S rRNA gene-based amplicon sequencing, were significantly different. The genera Lactobacillus, Weissella and Leuconostoc dominated in forage samples, whereas Pseudomonas, Staphylococcus and Streptococcus dominated in bulk milk samples. Forage quality and forage-associated microbiota were affected by ensiling method and by use of silage additive. Forages stored in bunker and tower silos (confounded with use of additive) were associated with higher levels of acetic and lactic acid and Lactobacillus. Forage ensiled as bales (confounded with no use of additive) was associated with higher dry matter content, water-soluble carbohydrate content, pH, yeast count and the genera Weissella, Leuconostoc and Enterococcus. For bulk tank milk samples, milking system was identified as the major factor affecting the microbiota and type of forage preservation had little impact. Analysis of common amplicon sequence variants (ASVs) suggested that forage was not the major source of Lactobacillus found in bulk tank milk.

Prevalence and spatial distribution of infectious diseases of dairy cattle in Ontario, Canada.

We investigated the prevalence and spatial distribution of selected pathogens associated with infectious diseases of dairy cattle in Ontario, Canada. The cross-sectional study surveyed bulk tank milk for antibodies against bovine leukemia virus (BLV), Mycobacterium avium ssp. paratuberculosis (MAP), and Salmonella Dublin, and for the presence of mastitis pathogens (Staphylococcus aureus, Streptococcus agalactiae, Mycoplasma bovis). Between October 2021 and June 2022, bulk tank milk samples were obtained from every commercial dairy farm in Ontario (n = 3,286). Samples underwent ELISA testing for the presence of BLV, MAP, and S. Dublin antibodies, and quantitative PCR testing for the detection of specific antigens of pathogens associated with mastitis. Bayesian models were used to estimate prevalence, and spatial analysis was carried out to identify regional clusters of high pathogen prevalence. Prevalence varied for different pathogens, and BLV was widespread across dairy farms in Ontario, with an estimated prevalence of 88.3%. The prevalence of MAP, Staph. aureus and S. Dublin in Ontario dairy herds was 39.8%, 31.5%, and 5.1%, respectively. The vast majority of dairy herds in Ontario were free of intramammary infections caused by Strep. agalactiae and M. bovis. Clusters of increased positive test rates were detected for S. Dublin, MAP, and Staph. aureus, indicating potential geographic risk factors for pathogen transmission. For S. Dublin, an area of increased test positivity rates was detected in southwestern Ontario, close to the Canada-United States border where most of the dairy herds in Ontario are located. Conversely, a localized cluster of positive test outcomes involving 14 subdivisions located in the southeastern region of Ontario was detected for Staph. aureus. Findings from our survey highlight the importance of the testing of aggregated samples and conducting spatial analysis as part of disease surveillance programs, and for implementing risk-based trading approaches among dairy producers.

Research on a Corrosion Detection Method for Oil Tank Bottoms Based on Acoustic Emission Technology.

This paper presents an acoustic emission (AE) detection method for refined oil storage tanks which is aimed towards specialized places such as oil storage tanks with high explosion-proof requirements, such as cave oil tanks and buried oil tanks. The method utilizes an explosion-proof acoustic emission instrument to detect the floor of a refined oil storage tank. By calculating the time difference between the defective acoustic signal and the speed of acoustic wave transmission, a mathematical model is constructed to analyze the detected signals. An independent channel AE detection system is designed, which can store the collected data in a piece of independent explosion-proof equipment, and can analyze and process the data in a safe area after the detection, solving the problems of a short signal acquisition distance and the weak safety protection applied to traditional AE instruments. A location analysis of the AE sources is conducted on the bottom plate of the tank, evaluating its corrosion condition accurately. The consistency between the evaluation and subsequent open-tank tests confirms that using AE technology effectively captures corrosion signals from oil storage tanks' bottoms. The feasibility of carrying out online inspection under the condition of oil storage in vertical steel oil tanks was verified through a comparison with open inspections, which provided a guide for determining the inspection target and opening order of large-scale oil tanks.

A Novel Experimental Apparatus for Characterizing Flow Regime in Mechanically Stirred Tanks through Force Sensors.

Pressure fluctuations in a mixing tank can provide valuable information about the existing flow regime within the tank, which in turn influences the degree of mixing that can be achieved. In the present work, we propose a prototype for identifying the flow regime in mechanically stirred tanks equipped with four vertical baffles through the characterization of pressure fluctuations. Our innovative proposal is based on force sensors strategically placed in the baffles of the mixing tank. The signals coming from the sensors are transmitted to an electronic module based on an Arduino UNO development board. In the electronic module, the pressure signals are conditioned, amplified and sent via Bluetooth to a computer. In the computer, the signals can be plotted or stored in an Excel file. In addition, the proposed system includes a moving average filtering and a hierarchical bottom-up clustering analysis that can determine the real-time flow regime (i.e., the Reynolds number, Re) in which the tank was operated during the mixing process. Finally, to demonstrate the versatility of the proposed prototype, experiments were conducted to identify the Reynolds number for different flow regimes (static, laminar, transition and turbulent), i.e., 0≤Re≤ 42,955. Obtained results were in agreement with the prevailing consensus on the onset and developed from different flow regimes in mechanically stirred tanks.

Design and Experiment of an Unoccupied Control System for a Tracked Grain Vehicle.

In order to enhance crop harvesting efficiency, an automatic-driving tracked grain vehicle system was designed. Based on the harvester chassis, we designed the mechanical structure of a tracked grain vehicle with a loading capacity of 4.5 m3 and a grain unloading hydraulic system. Using the BODAS hydraulic controller, we implemented the design of an electronic control system that combines the manual and automatic operation of the chassis walking mechanism and grain unloading mechanism. We utilized a hybrid A* algorithm to plan the traveling path of the tracked grain vehicle, and the path-tracking controller of the tracked grain vehicle was designed by combining fuzzy control and pure pursuit algorithms. Leveraging binocular vision technology and semantic segmentation technology, we designed an automatic grain unloading control system with functions of grain tank recognition and grain unloading regulation control. Finally, we conducted experiments on automatic grain unloading control and automatic navigation control in the field. The results showed that both the precision of the path-tracking control and the automatic unloading system meet the requirements for practical unoccupied operations of the tracked grain vehicle.

Risk Factors Associated with Musculoskeletal Injuries within the Crew of the Leopard 2 A6 Main Battle Tank Using Inertial Movement Unit Sensors: A Pilot Study.

This study aims to assess the musculoskeletal risk of military personnel on a Leopard 2 A6 main battle tank crew and to identify associated factors for future prevention and mitigation strategies. A sample of 57 Portuguese military personnel, who are or were part of the Leopard 2 A6 main battle tank crew, answered a questionnaire on their perception of task performance, considering muscle demands, comfort, posture, movements, and associated symptoms. A subsample of four soldiers from the Armoured Squadron of the Portuguese Mechanized Brigade were assessed using an inertial measurement unit system and underwent a whole-body kinematic analysis coupled with a Rapid Entire Body Assessment during a simulated two-hour mission. The results indicate that soldiers accurately perceive their roles within the crew and that, overall, there is a high risk of musculoskeletal injuries in all tasks. However, tasks directly related to the crew's primary duties carry consistently high risk when considering the time spent on their tasks. This study highlights the need for targeted preventive measures to reduce the incidence and severity of injuries among the crew of the Leopard 2 A6 main battle tank.

Coupling simulation of pipeline nodes - Storage tank linkage in urban high-density built-up areas using optimization model.

Climate change and urbanization contribute to the increased frequency of short-duration intense rainstorms. Traditional solutions often involve multiple scenarios for cost-effectiveness comparison, neglecting the rationality of placement conditions. The effective coupling and coordination of the location, number, size, and cost of storage tanks are crucial to addressing this issue. A three-phase approach is proposed to enhance the dynamic link between drainage pipeline and storage tanks in urban high-density built-up areas, integrating Python language, SWMM, the Elitist Non-Dominated Sorting Genetic Algorithm (NSGA-III), and the Analytic Hierarchy Process (AHP) methods. In the first stage, each node within the pipeline network is considered as a potential storage tank location. In the second stage, factors such as the length and diameter of the upstream connecting pipeline, as well as the suitability of the storage tank location, are assessed. In the third stage, the length and diameter of the downstream connecting pipeline node are evaluated. The results show that the 90 overflow nodes (overflow time >0.5h) have been cleared using the three-phase approach with a 50a (duration = 3h) return period as the rainfall scenario, which meets the flooding limitations. After the completion of the three-phase method configuration, the total overflow and SS loads were reduced by 96.45% and 49.30%, respectively, compared to the status quo conditions. These two indicators have decreased by 48.16 and 9.05%, respectively, compared to the first phase (the traditional method of only replacing all overflow nodes with storage tanks). The proposed framework enables decision-makers to evaluate the acceptability and reliability of the optimal management plan, taking into account their preferences and uncertainties.

Modulation of growth performance, feed utilization, and physiological traits in redbelly Tilapia (Tilapia zillii) through environmental and sex-based interactions.

This study explored the combined influence of tank color, stocking density, and gender on Tilapia zillii's performance and well-being. In this 120-day trial, 320 T. zillii, each initially weighing 10.0 ± 0.1 g/fish, were distributed among 24 tanks. The experiment included eight distinct treatment combinations, varying tank color (blue and green), stocking density (10 and 30 fish/m3) and sex (monosexual and mixed). The results showed that blue tanks improved specific growth rate and condition factor, while green tanks were better for feed utilization. Density at 30 fish/m3 showed the highest mean values of final body weight and total length, weight gain (WG), and gain length. Mono-sex outperformed mixed-sex ones in WG and daily growth. Interactions between color, density and sex were significant, affecting growth and feed utilization. Green tanks were best for protein profiles, while blue tanks excelled in glucose. A density of 10 fish/m3 yielded the highest protein profiles, and mono-sex fish had higher protein profiles. For lipid profiles, green tanks were superior, and density affected lipid profiles. Mixed-sex populations were best for certain lipid profile parameters. Interactions between these factors also played a significant role, making the biochemical profiles of T. zillii a complex interplay of various factors. The results explored that tank color, fish density and sex influence the activity of nonspecific immune enzymes in the liver of T. zillii. Blue tanks and lower fish density led to higher nonspecific immune enzymes, while mono-sex fish exhibited more significant nonspecific immune enzymes. Complex interactions between these factors also influenced nonspecific immune enzyme activities. Blue tanks increased malondialdehyde (MDA) levels, while green tanks raised glutathione S-transferases (GST) and catalase (CAT) levels. Lower fish density led to higher MDA, while higher density increased GST and CAT. Mono-sex fish had more MDA and GST, while mixed-sex fish showed greater CAT levels. Complex interactions among these factors affected the antioxidant levels in T. zillii. In summary, our study suggests that rearing T. zillii in green tanks at higher densities (30 fish/m3) and in mono-sex conditions yields the best results in terms of growth and overall performance.

The quest for down scale representativeness: how to exploit CFD to design a shear study for vaccines.

In this work, we exploit computational fluid dynamics (CFD) to evaluate stirred tank reactor (STR) process engineer parameters (PEP) and design a scale-down system (SDS) to be representative of the formulation and filling process steps for an Aluminum adjuvanted vaccine drug product (DP). To study the shear history in the SDS we used the concept of number of passages, combined with an appropriate stirring speed down scale strategy comprising of either (i) tip speed equivalence, widely used as a scale-up criterion for a shear-sensitive product, or (ii) rotating shear, a shear metric introduced by Metz and Otto in 1957 but never used as scaling criterion. The outcome of the CFD simulations shows that the tip equivalence generates a worst-case SDS in terms of shear, whereas the rotating shear scaling approach could be used to design a more representative SDS. We monitored the trend over time for "In Vitro Relative Potency" as DP Critical Quality Attribute for both scaling approaches, which highlighted the crucial role of choosing the appropriate scaling-down approach to be representative of the manufacturing scale during process characterization studies.