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The efficiency of direct electron flow from electron donors to electron acceptors in redox reactions is significantly influenced by the spatial separation of these components. Geobatteries, a class of redox-active substances naturally present in soil-water systems, act as electron reservoirs, reversibly donating, storing, and accepting electrons. This capability allows the temporal and spatial decoupling of redox half-reactions, providing a flexible electron transfer mechanism. In this review, we systematically examine the critical role of geobatteries in influencing electron transfer and utilization in environmental biogeochemical processes. Typical redox-active centers within geobatteries, such as quinone-like moieties, nitrogen- and sulfur-containing groups, and variable-valent metals, possess the potential to repeatedly charge and discharge. Various characterization techniques, ranging from qualitative methods like elemental analysis, imaging, and spectroscopy, to quantitative techniques such as chemical, spectroscopic, and electrochemical methods, have been developed to evaluate this reversible electron transfer capacity. Additionally, current research on the ecological and environmental significance of geobatteries extends beyond natural soil-water systems (e.g., soil carbon cycle) to engineered systems such as water treatment (e.g., nitrogen removal) and waste management (e.g., anaerobic digestion). Despite these advancements, challenges such as the complexity of environmental systems, difficulties in accurately quantifying electron exchange capacity, and scaling-up issues must be addressed to fully unlock their potential. This review underscores both the promise and challenges associated with geobatteries in responding to environmental issues, such as climate change and pollutant transformation.

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Methane fluxes (FCH4) vary significantly across wetland ecosystems due to complex mechanisms, challenging accurate estimations. The interactions among environmental drivers, while crucial in regulating FCH4, have not been well understood. Here, the interactive effects of six environmental drivers on FCH4 were first analyzed using 396,322 half-hourly measurements from 22 sites across various wetland types and climate zones. Results reveal that soil temperature, latent heat turbulent flux, and ecosystem respiration primarily exerted direct effects on FCH4, while air temperature and gross primary productivity mainly exerted indirect effects by interacting with other drivers. Significant spatial variability in FCH4 regulatory mechanisms was highlighted, with different drivers demonstrated varying direct, indirect, and total effects among sites. This spatial variability was then linked to site-specific annual-average air temperature (17.7%) and water table (9.0%) conditions, allowing the categorization of CH4 sources into four groups with identified critical drivers. An improved estimation approach using a random forest model with three critical drivers was consequently proposed, offering accurate FCH4 predictions with fewer input requirements. By explicitly accounting for environmental interactions and interpreting spatial variability, this study enhances our understanding of the mechanisms regulating CH4 emissions, contributing to more efficient modeling and estimation of wetland FCH4.

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The characteristics and dynamics of micro-plastisphere biofilm on the surface of microplastics (MPs) within artificial ecosystems, such as constructed wetlands (CWs), remain unclear, despite these ecosystems' potential to serve as sinks for MPs. This study investigates the dynamic evolution of micro-plastisphere biofilm in CWs, utilizing simulated wastewater containing sulfamethoxazole and humic acid, through physicochemical characterization and metagenomic analysis. Two different types of commercial plastics, including non-degradable polyethylene and degradable polylactic acid, were shredded into MPs and studied. The findings reveal that the types, shape and incubation time of MPs, along with humic acid content in wastewater, affected the quantity and quality of biofilms, such as the biofilm composition, spatial structure and microbial communities. After just 15 days into incubation, numerous microbials were observed on MP samples, with increases in biofilms content and enhanced humification of extracellular polymeric substances over time. Additionally, microbial communities on polylactic acid MPs, or those incubated for longer time, exhibit higher diversity, connectivity and stability, along with reduced vulnerability. Conversely, biofilms on polyethylene MPs were thicker, with higher potential for greenhouse gas emission and increased risk of antibiotic resistance genes. The addition of humic acid demonstrated opposite effects on biofilms across environmental interfaces, possibly due to its dual potential to produce light-induced free radicals and serve as a carbon source. Binning analysis further uncovered a unique assembly pattern of nutrients cycle genes and antibiotic resistance genes, significantly correlated within micro-plastisphere microbial communities, under the combined stress of nutrition and sulfamethoxazole. These results emphasize the shaping of micro-plastisphere biofilm characteristics by unique environmental conditions in artificial ecosystems, and the need to understand how DOM and other pollutants covary with MP pollution.

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OBJECTIVE: Analyze the impact of hyperbaric oxygen therapy on neuroprotection and recovery post severe traumatic brain injury (sTBI) resuscitation. METHODS: Retrospective analysis of clinical data from 83 sTBI patients admitted between January 2022 to January 2024. Patients were divided into control (n = 41) and observation (n = 42) groups based on treatment received. Control received standard therapy, while the observation group received hyperbaric oxygen therapy. Effects on clinical outcomes, neuroinjury markers (S100ß, GFAP, UCH-L1, NSE), neurotrophic factors (NGF, BDNF), neurological function indicators (NIHSS, CSS), and adverse reactions were compared. RESULTS: The observation group showed a higher total effective rate (80.95%) compared to control (60.98%) (p < 0.05). Neuroinjury markers decreased post-treatment in both groups, with the observation group lower (p < 0.05). NGF and BDNF levels increased post-treatment in both groups, with the observation group higher (p < 0.05). NIHSS and CSS scores decreased post-treatment in both groups, with the observation group lower (p < 0.05). No significant difference in adverse reactions between groups (p > 0.05). CONCLUSION: Hyperbaric oxygen therapy effectively treats sTBI by improving brain resuscitation success, reducing neuroinjury factors, enhancing neurotrophic factors, and promoting neurological function recovery, without increasing adverse reaction risk.

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This study was conducted to examine the efficacy of a bromelain-based supplementation coded ANR-pf on growth performance and intestinal lesion of broiler chickens under necrotic enteritis (NE) challenge. A total of 540 Ross 308 day-old male chicks were randomly allocated into 6 treatments of 6 replicates. The bromelain formulation was delivered to chickens through gavaging or in drinking water method twice, on d 8 and 13. Nonchallenged groups included 1) without or 2) with the specific bromelain formulation gavaged at 0.8 mL/kg. NE-challenged groups included 3) without the specific bromelain formulation; 4) gavaged with 0.4 mL/kg; 5) gavaged with 0.8 mL/kg and 6) supplemented with 0.8 mL/kg via drinking water. Birds were challenged with Eimeria spp. on d 9 and Clostridium perfringens (NE-18 strain) on d 14 and 15. On d 14 and 19, fresh faecal contents were collected for the determination of oocyst counts. Intestinal lesion scores were determined on d16. Performance and mortality were recorded throughout the entire experiment. Among challenged groups, birds received additive via drinking water had higher weight gain (WG) compared to the remaining groups (P < 0.001) in the grower phase and had lower FCR compared to 0.4 mL/kg inoculated group in the grower and finisher phases (P < 0.001). Bromelain supplementation via drinking water improved the WG of challenged birds, similar to that of the nonchallenged birds (P < 0.001), and lowered FCR compared to other challenged groups (P < 0.001). Nonchallenged birds and birds that received bromelain formulation in drinking water did not have lesions throughout the small intestine whereas challenged birds, either un-supplemented or supplemented with bromelain via inoculation route recorded similar lesion score levels in the jejunum. At d 19, birds received bromelain in drinking water had lower fecal oocyst numbers compared to challenged birds without additive (P < 0.001). In conclusion, bromelain administration via drinking water could ameliorate the negative impacts of NE-infection in broilers by improving performance, lowering the oocyst numbers and lesion scores.

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Stop codon readthrough (SCR) has important biological implications but remains largely uncharacterized. Here, we identify 1,009 SCR events in plants using a proteogenomic strategy. Plant SCR candidates tend to have shorter transcript lengths and fewer exons and splice variants than non-SCR transcripts. Mass spectrometry evidence shows that stop codons involved in SCR events can be recoded as 20 standard amino acids, some of which are also supported by suppressor tRNA analysis. We also observe multiple functional signals in 34 maize extended proteins and characterize the structural and subcellular localization changes in the extended protein of basic transcription factor 3. Furthermore, the SCR events exhibit non-conserved signature, and the extensions likely undergo protein-coding selection. Overall, our study not only characterizes that SCR events are commonly present in plants but also identifies the recoding plasticity of stop codons, which provides important insights into the flexibility of genetic decoding.

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This study aimed to investigate the protective effects and mechanisms of hyperbaric oxygen (HBO) preconditioning in a rat model of acute myocardial infarction (MI) established by ligation of the left anterior descending (LAD) coronary artery. Microarray, real-time PCR, and western blotting (WB) results demonstrated that the Mst1 gene was downregulated in the heart tissue of the MI rat model. HBO preconditioning significantly increased Mst1 expression in cardiac tissues of rats after MI modeling. Lentiviral infection was used to silence the Mst1 gene in rats treated with HBO to probe the effect of Mst1 on HBO cardioprotection. HBO preconditioning decreased heart infarct size and ameliorated cardiac function in MI rats, whereas Mst1 silencing reversed the effect of HBO administration, as indicated after heat infarct size determination via TTC staining, histological examination via HE staining, and measurements of cardiac function. HBO preconditioning reduced oxidative stress and inflammation in cardiac tissue of MI rat model, evidenced by alteration of malondialdehyde (MDA), 8-hydroxy-2-deoxyguanosine (8-OHdG), and protein carbonyl contents, as well as production of inflammation-associated myeloperoxidase (MPO), IL-1ß, and TNF-α. These findings provide a new signaling mechanism through which HBO preconditioning can protect against acute MI injury through the Mst1-mediating Keap1/Nrf2/HO-1-dependent antioxidant defense system.

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Different energy systems have been proposed for energy evaluation of feeds for domestic animals. The oldest and most commonly used systems take into account the fecal energy loss to obtain digestible energy (DE), and fecal, urinary and fermentation gases energy losses to calculate metabolizable energy (ME). In the case of ruminants and pigs, the net energy (NE) system, which takes into account the heat increment associated with the metabolic utilization of ME, has progressively replaced the DE and ME systems over the last 50 years. For poultry, apparent ME (AME) is used exclusively and NE is not yet used widely. The present paper considers some important methodological points for measuring NE in poultry feeds and summarizes the available knowledge on NE systems for poultry. NE prediction equations based on a common analysis of three recent studies representing a total of 50 complete and balanced diets fed to broilers are proposed; these equations including the AME content and easily available chemical indicators have been validated on another set of 30 diets. The equations are applicable to both ingredients and complete diets. They rely primarily on an accurate and reliable AME value which then represents the first limiting predictor of NE value. Our analysis indicates that NE would be a better predictor of broiler performance than AME and that the hierarchy between feeds is dependent on the energy system with a higher energy value for fat and a lower energy value for protein in an NE system. Practical considerations for implementing such an NE system from the commonly used AME or AMEn (AME adjusted for zero nitrogen balance) systems are presented. In conclusion, there is sufficient information to allow the implementation of the NE concept in order to improve the accuracy of feed formulation in poultry.

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It is well known that male and female broilers differ in their growth performance and that many physiological factors contribute to this difference. The aim of this experiment is to investigate if there are differences between male and female broilers in cecal microbiota and nutrient transporter gene expression and if these differences play a role in the growth performance of broilers. The possible effect of protein level and its interaction with sex on microbiota and expression of the nutrient transporters were also investigated. Samples were collected from male and female birds fed either standard crude protein (SCP) or reduced crude protein diets (RCP) at the age of d 35. The experiment was designed as a 2 × 2 factorial arrangement of treatments consisting of 448 Cobb 500 broilers assigned to 32-floor pens with 4 treatments, 8 replicates, and 14 birds per pen for performance measurements. The factors were sex (male or female) and dietary crude protein (CP) level (standard or reduced). Body weight gain (BWG), feed intake and feed conversion ratio were recorded for each pen. Sex had a significant effect on BWG and FCR (P < 0.001) where males had a significantly higher BWG and better FCR compared to females. There was a significant interaction between sex and protein level on feed intake (FI) (P < 0.05), where male birds had a higher FI compared to female birds only when the birds were fed SCP but not RCP diets. There was a significant interaction between CP level and sex on the expression of CAT2 (P = 0.02) and PEPT2 (P = 0.026) where the genes were significantly upregulated in females but only when the RCP diet was fed. The RCP diet upregulated the expression of BoAT (P = 0.03) as a main effect. Female birds had significantly higher expression of the PepT-2 gene compared to the males. The alpha diversity of the cecal microbiota showed differences among the treatments. The Shannon diversity index was statistically higher (P = 0.036) for males fed the SCP diet and the Chao1 index for evenness was statistically higher (P = 0.027) in females fed the SCP diet. There was also a difference in the relative abundance of the 15 most common genera found in the cecal content of the broilers in this experiment and lastly, the differential composition of microbiota between the different treatments was also significantly different. This study suggests that chickens are able to compensate for a reduction in AA substrates when fed a low CP diet through the upregulation of certain AA transporters, females may adapt to low CP diets better by such upregulation compared to males, and lastly, sex has an effect on the cecal microbial population and these differences contribute towards the performance differences between male and female broilers.

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Background: The Black Soldier Fly larvae (BSFL) are a source of nutrients and bioactive compounds in broiler diets. Some components of the BSFL may serve as a prebiotic or may impact the intestinal microbiota of the broilers by other modes of action, which in turn can affect the health and performance of broilers. Here, we investigate the impact of up to 20% BSFL in broiler diets on the diversity and composition of the broiler's microbiota. Methods: Four hundred broilers were fed five iso-nutritious experimental diets with increasing levels of BSFL meal reaching 0%, 5%, 10%, 15%, 20% in the finisher diets. Eight caecal content samples coming from each of the eight replicates per treatment were collected at two time points (day 21 and day 42) for DNA extraction and sequencing of the V3-V4 regions using Illumina MiSeq 2 × 300 bp pair-end sequencing with 341f and 805r primers. Analysis of variance and Spearman's correlation were performed, while QIIME2, DADA2, and Calypso were used for data analysis. Results: When broilers were 21 days of age, the abundance of two groups of sequence variants representing Enterococcus and unclassified Christensenellaceae was significantly lower (p-value = 0.048 and p-value = 0.025, respectively) in the 20% BSFL group compared to the 0% BSFL group. There was no relevant alteration in the microbiota diversity at that stage. On day 42, the Spearman correlation analysis demonstrated that the sequence variants representing the genus Coprococcus showed a negative relationship with the BSFL inclusion levels (p-value = 0.043). The sequence variants representing the genus Roseburia and Dehalobacterium demonstrated a positive relationship with the BSFL dietary inclusion (p-value = 0.0069 and p-value = 0.0034, respectively). There was a reduction in the dissimilarity index (ANOSIM) caused by the 20% BSFL dietary inclusion. Conclusion: The addition of up to 20% BSFL in broiler diets did not affect the overall caeca microbiota diversity or composition at day 21. On day 42, there was a reduction in the beta diversity caused by the 20% BSFL dietary inclusion. The abundance of the bacterial group Roseburia was increased by the BSFL dietary inclusion, and it may be beneficial to broiler immunity and performance.

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Coccidiosis is an enteric disease of poultry worldwide that compromises gut health and growth performance. The current research investigated the effects of 2 doses of a multienzyme preparation on broilers' performance, gut health, and footpad dermatitis (FPD) under an Eimeria challenge. A total of 512 mixed-sex day-old chicks (Cobb 500) were randomly allocated to 4 treatments of 8 replicates. Treatments were: 1) nonchallenged control (NC); 2) NC + Eimeria challenge (CC); 3) CC + recommended level of xylanase and glucanase (XG, 100 g/t feed [on top]); 4) CC + double XG (2XG, 200 g/t feed). Eimeria spp. vaccine strains were gavaged on d 9 to induce coccidiosis in chickens. Performance parameters were evaluated during starter, grower, and finisher phases, and 4 birds per pen were euthanized on d 16 for sampling, FPD was scored on d 35, and litter moisture was analyzed on d 17 and 35. The data were analyzed using 1-way ANOVA with Tukey's test to separate means, and Kruskal-Wallis test was used for non-normally distributed parameters. The results showed that the Eimeria challenge was successful based on reduced weight gain and feed intake during grower phase, and higher FITC-d concentration, lesion score (female), and oocyst counts (d 14) in CC group compared to N.C. group, while XG and 2XG increased (P < 0.001) weight gain and improved FCR compared to CC and NC groups during finisher phase. The addition of X.G. and 2XG decreased litter moisture (P = 0.003) and FPD (P < 0.001) in challenged broilers compared to the N.C. group (d 35). Supplementing XG and 2XG reestablished the population of Lactobacillus in the cecum of challenged birds to an intermediate level between the NC and CC groups (P > 0.05). The inclusion of XG tended to increase the expression of Junctional adhesion molecule 2 (JAM2), which was not different from CC and NC groups (P > 0.05). In conclusion, the combination of xylanase and glucanase (Natugrain TS) improved the performance and modulated jejunal microbiota of broilers under mild Eimeria challenge.

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Due to the removal of antibiotics from animal feed, alternatives have been sought to control necrotic enteritis (NE) in broilers. The current study investigated the effects of buffered formic acid (Amasil NA) and monoglycerides of short- and medium-chain fatty acids (Balangut LS P) on the performance and gut health of broilers challenged with NE. A total of 816 as-hatched 1-d-old chicks (Cobb 500) were randomly assigned to 6 treatments with 8 replicates. Treatments were: T1) nonchallenged control; T2) NE challenged control; T3) Amasil NA (challenge plus Amasil NA, 0.3% throughout all phases); T4) Balangut LS P (challenge plus Balangut LS P, 0.5%, 0.3%, and 0.2% in the starter, grower and finisher phases, respectively; T5) Combined (challenge plus combination of T3 and T4); T6) Antibiotic (challenge plus Zn bacitracin, 0.05 % throughout all phases). Birds were orally gavaged with live Eimeria vaccine species (d 9) and with Clostridium perfringens (d 14 and 15). On d 16, birds were sampled to evaluate gut permeability, microbiota, and mRNA abundance in the jejunum. The data were analyzed in JMP software using one-way ANOVA with Tukey's test to separate means, and Kruskal-Wallis test was used for non-normally-distributed parameters. Results showed that Balangut LS P decreased (P<0.05) feed conversion ratio compared to nonchallenged ones at the end of the study. Balangut LS P reduced (P < 0.05) the level of cecal Bacteriods compared to nonchallenged group, whereas Amasil NA shifted the levels of ileal Bifidobacteria, Enterobacteriaceae, and Lactobacillus towards nonchallenged control (P > 0.05). NE challenge upregulated (P < 0.001) the expression of IL-21R, zeta chain of T cell receptor (ZAP70), and dual specificity phosphatase 4 (DUSP4) compared to nonchallenged birds, whereas Balangut LS P showed an intermediate (P > 0.05) expression pattern of these genes towards nonchallenged and antibiotic groups. In conclusion, combination of Balangut LS P and Amasil NA has the potential to be used as an additive to improve the performance and gut health of broiler chickens, especially under challenging conditions such as NE infections.

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Artificial aeration is a widely used approach in wastewater treatment to enhance the removal of pollutants, however, traditional aeration techniques have been challenging due to the low oxygen transfer rate (OTR). Nanobubble aeration has emerged as a promising technology that utilise nano-scale bubbles to achieve higher OTRs owing to their large surface area and unique properties such as longevity and reactive oxygen species generation. This study, for the first time, investigated the feasibility of coupling nanobubble technology with constructed wetlands (CWs) for treating livestock wastewater. The results demonstrated that nanobubble-aerated CWs achieved significantly higher removal efficiencies of total organic carbon (TOC) and ammonia (NH4+-N), at 49 % and 65 %, respectively, compared to traditional aeration treatment (36 % and 48 %) and the control group (27 % and 22 %). The enhanced performance of the nanobubble-aerated CWs can be attributed to the nearly three times higher amount of nanobubbles (Ø < 1 µm) generated from the nanobubble pump (3.68 × 108 particles/mL) compared to the normal aeration pump. Moreover, the microbial fuel cells (MFCs) embedded in the nanobubble-aerated CWs harvested 5.5 times higher electricity energy (29 mW/m2) compared to the other groups. The results suggested that nanobubble technology has the potential to trigger the innovation of CWs by enhancing their capacity for water treatment and energy recovery. Further research needs are proposed to optimise the generation of nanobubbles, allowing them to be effectively coupled with different technologies for engineering implementation.

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Microplastics (MPs) are emerging pollutants that interact extensively with dissolved organic matter (DOM) and this influences the environmental behavior of MPs in aqueous ecosystems. However, the effect of DOM on the photodegradation of MPs in aqueous systems is still unclear. The photodegradation characteristics of polystyrene microplastics (PS-MPs) in an aqueous system in the presence of humic acid (HA, a signature compound of DOM) under ultraviolet light conditions were investigated in this study through Fourier transform infrared spectroscopy coupled with two-dimensional correlation analysis, electron paramagnetic resonance, and gas chromatography-mass spectrometry (GC/MS). HA was found to promote higher levels of reactive oxygen species (0.631 mM of ▪OH), which accelerated the photodegradation of PS-MPs, with a higher degree of weight loss (4.3%), higher level of oxygen-containing functional groups, and lower average particle size (89.5 µm). Likewise, GC/MS analysis showed that HA contributed to a higher content of oxygen-containing compounds (42.62%) in the photodegradation of PS-MPs. Moreover, the intermediates and final degradation products of PS-MPs with HA were significantly different in the absence of HA during 40 days of irradiation. These results provide an insight into the co-existing compounds on the degradation and migration processes of MP and also support further research toward the remediation of MPs pollution in aqueous ecosystems.

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It has been demonstrated that the fiber content of oilseed meals and total amino acids (AA) in distillers' grains products affects standardized ileal digestibility (SID) values from swine assays and that total protein concentration affects the AA digestibility in assays using chickens. This analysis was conducted to test the hypothesis that the total AA content of test samples affects the SID assay results. Databases containing total AA profiles and SID values of 20 feedstuffs commonly fed to pigs from 2 sources, AMINODat 5.0 (Evonik Industries, 2015) and the Nutrient Requirements of Swine, 12th edition (NRC, 2012) were used to compare AA concentration effects on standardized ileal digestibility coefficients. Databases were compared with AA and SID available in both data sets. The total AA values were similar for the 2 databases with an R 2 of 0.979 (P < 0.001). The linear relationship between digestibility coefficients from the 2 databases was highly significant, an R 2 of 0.810 (P < 0.001). Both databases had increased SID values with increasing AA contents within and across feed ingredients. The SID = f(AA concentration) relationship was confirmed with an individual paper. Since SID, as typically measured, is a function of both digestion and absorption, both processes following Michaelis-Menten kinetics, SID = f(AA concentration) may simply be a natural phenomenon. Other reasons for the relationship were explored. Methods of estimating endogenous AA losses and misapplication statistical procedures may contribute to variation in results and at least partially explain why SID = f(AA concentration).

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In this study, two constructed wetland-microbial fuel cells (CW-MFC), including a closed-circuit system (CCW-MFC) and an open-circuit system (OCW-MFC) with petroleum coke as electrode and substrate, were constructed to explore the effect of multiple key factors on their operation performances. Compared to a traditional CW, the CCW-MFC system showed better performance, achieving an average removal efficiency of COD, NH4+-N, and TN of 94.49 ± 1.81%, 94.99 ± 4.81%, and 84.67 ± 5.6%, respectively, when the aeration rate, COD concentration, and hydraulic retention time were 0.4 L/min, 300 mg/L, and 3 days. The maximum output voltage (425.2 mV) of the CCW-MFC system was achieved when the aeration rate was 0.2 L/min. In addition, the CCW-MFC system showed a greater denitrification ability due to the higher abundance of Thiothrix that might attract other denitrifying bacteria, such as Methylotenera and Hyphomicrobium, to participate in the denitrifying process, indicating the quorum sensing could be stimulated within the denitrifying microbial community.

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The slime mould algorithm (SMA) is a new meta-heuristic algorithm recently proposed. The algorithm is inspired by the foraging behavior of polycephalus slime moulds. It simulates the behavior and morphological changes of slime moulds during foraging through adaptive weights. Although the original SMA's performance is better than most swarm intelligence algorithms, it still has shortcomings, such as quickly falling into local optimal values and insufficient exploitation. This paper proposes a Gaussian barebone mutation enhanced SMA (GBSMA) to alleviate the original SMA's shortcomings. First of all, the Gaussian function in the Gaussian barebone accelerates the convergence while also expanding the search space, which improves the algorithm exploration and exploitation capabilities. Secondly, the differential evolution (DE) update strategy in the Gaussian barebone, using rand as the guiding vector. It also enhances the algorithm's global search performance to a certain extent. Also, the greedy selection is introduced on this basis, which prevents individuals from performing invalid position updates. In the IEEE CEC2017 test function, the proposed GBSMA is compared with a variety of meta-heuristic algorithms to verify the performance of GBSMA. Besides, GBSMA is applied to solve truss structure optimization problems. Experimental results show that the convergence speed and solution accuracy of the proposed GBSMA are significantly better than the original SMA and other similar products. Supplementary Information: The online version contains supplementary material available at 10.1007/s10462-022-10370-7.

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The availability of sexed day-old broiler chicks is becoming an issue as feather sexing is no longer possible. This has great implications for broiler researchers as the use of randomly distributed mixed-sex birds may result in a greater between-pen variation and thus less statistical power than the use of single-sex birds. The objective of this study was to evaluate the effect of including sex proportion as a covariate in an analysis of covariance (ANCOVA) on the statistical power compared to analysis of variance (ANOVA) where sex was not considered. The statistical parameters examined include mean square error (MSE), the F-statistic, model fit, model significance and observed power. A total of 4 separate experiments that used mixed-sex broilers with unequal numbers of male and female birds per pen were conducted during which performance of the birds was measured. The male % in each pen was recorded during each experiment and corrected for mortality. The performance results were analysed by ANOVA and the statistical parameters were then compared to ANCOVA where sex proportion was included as a covariate. The results showed that a set of assumptions first needed to be met to run ANCOVA. In addition, if the ANOVA results show a high level of model significance and power, then ANCOVA may not be necessary. In other circumstances where the assumptions are met and model significance and observed power are low, the inclusion of sex proportion as a covariate in the analysis will help to reduce MSE, increase the F-statistic value and improve the model significance, model fit and observed power. Therefore, it is suggested that sex proportion should be considered as a covariate in ANCOVA to improve statistical power in nutritional experiments when male and female broilers are unequally and randomly distributed in pens.

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