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Most microorganisms resist pure cultivation under conventional laboratory conditions. One of the primary issues for this un-culturability is the absence of biologically produced growth-promoting factors in traditionally defined growth media. However, whether cultivating microbes by providing spent culture supernatant of pivotal microbes in the growth medium can be an effective approach to overcome this limitation is still an under-explored area of research. Here, we used the spent culture medium (SCM) method to isolate previously uncultivated marine bacteria and compared the efficiency of this method with the traditional cultivation (TC) method. In the SCM method, Ca. Bathyarchaeia-enriched supernatant (10%) was used along with recalcitrant organic substrates such as lignin, humic acid, and organic carbon mixture. Ca. Bathyarchaeia, a ubiquitous class of archaea, have the capacity to produce metabolites, making their spent culture supernatant a key source to recover new bacterial stains. Both cultivation methods resulted in the recovery of bacterial species from the phyla Pseudomonadota, Bacteroidota, Actinomycetota, and Bacillota. However, our SCM approach also led to the recovery of species from rarely cultivated groups, such as Planctomycetota, Deinococcota, and Balneolota. In terms of the isolation of new taxa, the SCM method resulted in the cultivation of 80 potential new strains, including one at the family, 16 at the genus, and 63 at the species level, with a novelty ratio of ~ 35% (80/219). In contrast, the TC method allowed the isolation of ~ 10% (19/171) novel strains at species level only. These findings suggest that the SCM approach improved the cultivation of novel and diverse bacteria.

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Black soldier fly larvae (Hermetia illucens; BSFL) can transform organic wastes into nutritional biomass useful in animal feeding. The aim of this work was to study the effect of five diets (meat, fruit, vegetable substrates, a mix of them and control) on the profile of fatty acids (FAs) and sterols of BSFL. For a more exhaustive characterization of the nutritional properties, the profile of esterified FAs in the sn-2 position of the triglycerides, the most absorbed lipid component during animal digestion was evaluated. The dietary effect was estimated on two different Hermetia illucens populations (Greek - UTH and Italian - UNIPI). The diet affected all the lipid fractions examined. Regardless of diet, the fat was characterized mainly of lauric acid and other saturated FAs, which were found to be synthesized by the larvae, as it was not present in any of the five substrates. In general, UTH larvae contained a higher level of lipids (7.38 vs 2.48 g/100 g of larvae; P < 0.001) and saturated FAs (49.71 vs 36.10 g/100 g of Total Lipids; P < 0.001) and a lower percentage of monounsaturated FAs (14.74 vs 26.70 g/100 g of Total Lipids), C18:3n-3 (0.67 vs 1.13 g/100 g of Total Lipids; P < 0.001), and C18:2c9t11 (2.02 vs 2.80 g/100 g of Total Lipids; P < 0.001). Irrespective of the populations, BSFL reared on control and fruit substrates showed higher level of lipids (8.06 and 5.61 g/100 g of larvae, respectively), and saturated FA (38.99 and 71.19 g/100 g of Total Lipids, respectively), while the presence of meat increased the level of C20:4n-6, C20:5n-3 and C22:5n-3 (0.70, 0.13 and 0.45 g/100 g of Total Lipids, respectively). The results confirmed that BSFL accumulate phytosterols in their lipid fraction. The sterol profile was strongly influenced by the substrate on which the larvae were reared, with higher levels of cholesterol in the larvae of the meat group (38.55 mg/100 g of Total Lipids) and of stigmasterol and campesterol (9.04 and 15.23 mg/100 g of Total Lipids, respectively) in those of the vegetable group. The sterol content between the two populations was significantly different, with a higher percentage in UTH larvae (113.28 vs 34.03 mg/100 g of Total Lipids; P < 0.001). Finally, BSFLs showed a high plasticity of the lipid profile depending on both the substrate and the metabolism linked to the different populations. This variability allows the nutritional characteristics of the BSFL to be shaped by modifying the substrate, to adapt it to the technological and feeding needs to which the larvae are destined.

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Sclerotinia sclerotiorum (Lib.) de Bary, an economically devastating soilborne fungal pathogen known to cause disease across a wide range of plants, produces long-term inoculum called sclerotia that can germinate either carpogenically by ascospores infecting aboveground plant parts or myceliogenically to infect stem base and roots. Typically, for research purposes, S. sclerotiorum diseases are initiated by direct contact methods, using S. sclerotiorum mycelium agar plugs wrapped around the stem or sclerotia placed directly beneath root mass. However, reproducible noncontact methods leading to basal stem infection are not currently available. Therefore, the objective of this study was to develop effective noncontact protocols that consistently generate basal plant stem infection from S. sclerotiorum in the soil. Using three host plant species (canola, lupin, and lettuce), we determined two methods that reliably produced basal stem infection. The first method, where mycelial agar plugs were positioned just below the soil surface at a distance of 5 mm from each seedling, led to 100% infection in all plants. The second method used pathogen-infested soil by mixing the soil with dry inoculum in the form of a powder prepared from mycelium-colonized organic substrates. Four substrates consistently produced 100% seedling infection at 4 days after inoculation (DAI): wheat bran, wheat grain, red rice, and hulled millet. In contrast, chia, canary, sesame, and ryegrass seed substrates resulted in less than 50% seedling infection at 10 DAI, and infection levels did not progress further. The two soil inoculation methods outlined in this study will enhance future research on the progression of S. sclerotiorum diseases, with the potential to screen disease-resistant host genotypes to basal S. sclerotiorum infection and, in particular, to test the effectiveness of soil applications of fungicides or biocontrol agents against S. sclerotiorum basal infection.

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Assessment of wastewater effluent quality in terms of physicochemical and microbial parameters is a difficult task; therefore, an online method which combines the variables and represents a final value as the quality index could be used as a useful management tool for decision makers. However, conventional measurement methods often have limitations, such as time-consuming processes and high associated costs, which hinder efficient and practical monitoring. Therefore, this study presents an approach that underscores the importance of using both short- and long-term memory networks (LSTM) to enhance monitoring capabilities within wastewater treatment plants (WWTPs). The use of LSTM networks for soft sensor design is presented as a promising solution for accurate variable estimation to quantify effluent quality using the total chemical oxygen demand (TCOD) quality index. For the realization of this work, we first generated a dataset that describes the behavior of the activated sludge system in discrete time. Then, we developed a deep LSTM network structure as a basis for formulating the LSTM-based soft sensor model. The results demonstrate that this structure produces high-precision predictions for the concentrations of soluble X1 and solid X2 substrates in the wastewater treatment system. After hyperparameter optimization, the predictive capacity of the proposed model is optimized, with average values of performance metrics, mean square error (MSE), coefficient of determination (R2), and mean absolute percentage error (MAPE), of 23.38, 0.97, and 1.31 for X1, and 9.74, 0.93, and 1.89 for X2, respectively. According to the results, the proposed LSTM-based soft sensor can be a valuable tool for determining effluent quality index in wastewater treatment systems.

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The 'Criolla' potato (Solanum tuberosum) phureja group cultivated in the inter-Andean valleys of the high tropics, is commonly fertilized only with mineral nutrients in conventional production, without the use of soil test and use of organic fertilizers. The purpose of this study was to determine the potential of organic chicken manure as a source of nutrients to increase yield and quality in 'Criolla' potatoes compared to conventional fertilization. Treatments evaluated were organic chicken manure, mineral nutrition based on soil tests, organic manure mixed with mineral nutrition, and conventional nutrition based on farmers' typical nutrition plans. In plant response conventional and mineral treatments mixed with organic matter were the best in leaf area and dry weight of shoot and tuber. Treatment with mineral nutrition mixed with organic manure (2 and 6 t) obtained the largest length of the main stems. The best yield response was obtained with conventional nutrition, while the major size was obtained with mineral and mineral combined with organic treatments. According to the results, the best response was obtained with the application of mineral nutrition in the combination of 6 t ha-1 organic manure.

La papa criolla (Solanum tuberosum) grupo phureja es cultivada en los valles interandinos del trópico alto, comúnmente fertilizada con nutrientes minerales en la producción convencional, sin hacer uso de análisis de suelos y nutrición con fertilizantes orgánicos. El objetivo de este estudio fue determinar el potencial de la materia orgánica de gallinaza, como fuente de nutrientes, para incrementar el rendimiento y la calidad en la papa criolla, comparándola con la nutrición convencional. Los tratamientos evaluados fueron gallinaza, nutrición mineral, basado en análisis de suelos; nutrición mineral, combinada con materia orgánica y un tratamiento soportado en la fertilización convencional, con base en los planes nutricionales, típicos de los agricultores. En las respuestas de la planta, los tratamientos convencionales y minerales mezclados con nutrición mineral obtuvieron los valores más altos en área foliar y peso seco de la parte aérea y el tubérculo. En longitud de tallo, la mejor respuesta se obtuvo con los tratamientos minerales mezclados con materia orgánica (2 y 6 toneladas). La mejor respuesta en rendimiento fue obtenida en la nutrición mineral convencional, mientras que las papas con mayor calibre, se lograron con los tratamientos mineral y mineral mezclado con orgánico. De acuerdo con los resultados, la mejor respuesta en cultivo se puede obtener con la nutrición mineral, en combinación con 6 toneladas de materia orgánica.

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Considerable advances have been made in the substrate design and operation strategies of constructed wetlands to facilitate nitrogen elimination. However, few studies examined the complicated interaction between solid organic substrates and limited aeration on nitrogen removal. A vertical flow constructed wetlands in gradient distribution of inorganic and solid organic substrates (polycaprolactone/PCL) (P-VFCW) and a controlled vertical flow constructed wetland without PCL (C-VFCW) were developed for the tertiary treatment of municipal tailwater. Results indicated that ammonia was nearly converted to nitrate, while the total nitrogen removal efficiencies (TNREs) in C-VFCW were negligible. In P-VFCW, however, optimal TNREs approached 95% with an aeration rate of 0.06 mL·min-1 and hydraulic retention time (HRT) of 24 h, and simultaneous nitrification and denitrification process (SND) in aerobic conditions was confirmed. As for the spatial microbial community structure evolution, Comamonas, which is associated with heterotrophic nitrification and anoxic/aerobic denitrification, was enriched along the vertical profiles of P-VFCW. Autotrophic nitrifier (Nitrospira), aerobic denitrifier (Bradyrhizobium and Azospira), and anoxic denitrifier (Ignavibacterium and Methyloversatilis) were dominated in different depths of P-VFCW, respectively. Besides, Canna indica biomass in P-VFCW was significantly larger than that in C-VFCW, which was attributed to the plant adaption response to diverse nitrogen. The P-VFCW in gradient distribution of inorganic and solid carbon sources under limited aeration is a promising technology for advanced nitrogen removal.

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Microbial electrosynthesis system (MES) is a promising method that can use carbon dioxide, which is a greenhouse gas, to produce methane which acts as an energy source, without using organic substances. However, this bioelectrical reduction reaction can proceed at a certain high applied voltage when coupled with water oxidation in the anode coated with metallic catalyst. When coupled with the oxidation of HS- to SO42-, methane production is thermodynamically more feasible, thus implying its production at a considerably lower applied voltage. In this study, we demonstrated the possibility of electrotrophic methane production coupled with HS- oxidation in a cost-effective bioanode chamber in the MES without organic substrates at a low applied voltage of 0.2 V. In addition, microbial community analyses of biomass enriched in the bioanode and biocathode were used to reveal the most probable pathway for methane production from HS- oxidation. In the bioanode, electroautotrophic SO42- production accompanied with electron donation to the electrode is performed mainly by the following two steps: first, incomplete sulfide oxidation to sulfur cycle intermediates (SCI) is performed; then the produced SCI are disproportionated to HS- and SO42-. In the biocathode, methane is produced mainly via H2 and acetate by electron-accepting syntrophic bacteria, homoacetogens, and acetoclastic archaea. Here, a new eco-friendly MES with biological H2S removal is established.

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BACKGROUND: Commonly, several traits are assessed in agronomic experiments to better understand the factors under study. However, it is also common to see that even when several traits are available, researchers opt to follow the easiest way by applying univariate analyses and post-hoc tests for mean comparison for each trait, which arouses the hypothesis that the benefits of a multi-trait framework analysis may have not been fully exploited in this area. RESULTS: In this paper, we extended the theoretical foundations of the multi-trait genotype-ideotype distance index (MGIDI) to analyze multivariate data either in simple experiments (e.g., one-way layout with few treatments and traits) or complex experiments (e.g., with a factorial treatment structure). We proposed an optional weighting process that makes the ranking of treatments that stands out in traits with higher weights more likely. Its application is illustrated using (1) simulated data and (2) real data from a strawberry experiment that aims to select better factor combinations (namely, cultivar, transplant origin, and substrate mixture) based on the desired performance of 22 phenological, productive, physiological, and qualitative traits. Our results show that most of the strawberry traits are influenced by the cultivar, transplant origin, cultivation substrates, as well as by the interaction between cultivar and transplant origin. The MGIDI ranked the Albion cultivar originated from Imported transplants and the Camarosa cultivar originated from National transplants as the better factor combinations. The substrates with burned rice husk as the main component (70%) showed satisfactory physical proprieties, providing higher water use efficiency. The strengths and weakness view provided by the MGIDI revealed that looking for an ideal treatment should direct the efforts on increasing fruit production of Albion transplants from Imported origin. On the other hand, this treatment has strengths related to productive precocity, total soluble solids, and flesh firmness. CONCLUSIONS: Overall, this study opens the door to the use of MGIDI beyond the plant breeding context, providing a unique, practical, robust, and easy-to-handle multi-trait-based framework to analyze multivariate data. There is an exciting possibility for this to open up new avenues of research, mainly because using the MGIDI in future studies will dramatically reduce the number of tables/figures needed, serving as a powerful tool to guide researchers toward better treatment recommendations.

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Bacteria play a key role in biogeochemical cycles in natural and anthropogenic ecosystems. In river ecosystems, bacteria intensively colonize silt sediments. Microorganisms are essential for energy conversion, biogeochemical nutrient cycling, pollutant degradation, and biotransformation of organic matter; therefore, bottom sediments can be a source of metabolically diverse microorganisms, including those with promise for industrial biotechnologies. The aim of this work was to isolate and study pure cultures of microorganisms - producers of industrially important enzymes and decomposers of organic matter - from bottom sediments of the Ob River. Pork fat and diesel fuel were used as substrates to obtain enrichment and pure cultures for selective cultivation of bacteria with lipolytic and hydrocarbon-oxidizing activity. A total of 21 pure cultures were isolated. The phylogenetic position of the obtained bacterial isolates was determined based on the analysis of 16S rRNA gene sequences. The strains isolated on selective media belonged to representatives of the genera Pseudomonas and Aeromonas (Gammaproteobacteria), and the genus Microvirgula (Betaproteobacteria). The ability of strains to grow on culture media containing pork fat, olive oil and diesel fuel was analyzed. The lipolytic activity of the isolates was evidenced by cultivation on a diagnostic medium containing 1 % tributyrin. The phylogenetic and metabolic diversity of the cultivated non-pathogenic bacterial strains with lipolytic and oil-oxidizing activity revealed in the study indicates the biotechnological potential of the isolates. The most promising strains were M. aerodenitrificans sp. LM1 and P. lini sp. KGS5K3, which not only exhibited lipolytic activity on the diagnostic medium with tributyrin in a wide temperature range, but also utilized diesel fuel, pork fat and olive oil.

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Passiflora setacea DC. growing is of interest to the herbal industries since in its leaves are produced secondary metabolites that confer antioxidant, anxiolytic, and antidepressant properties in Passiflora. Therefore, it is important to search for sustainable alternatives that aim to enhance the production of these compounds to add value to the phytomass, such as the inoculation with arbuscular mycorrhizal fungi (AMF) and the application of coconut coir dust, which has not been reported to P. setacea yet. The aim was to select the efficient combination of AMF and coconut coir dust to increase the compounds' production and optimize the antioxidant activity in P. setacea leaves. The P. setacea seedlings that were cultivated in substrates without coconut coir dust and colonized by Gigaspora albida N.C. Schenck & G.S. Sm. produced more total saponins (1,707.43%), total tannins (469.98%), and total phenols (85.81%), in comparison to the non-mycorrhizal plants, in addition to enhancing the glomalin-related soil proteins. On the other hand, in general, the use of coir dust as a substrate has not been shown to increase the production of these bioactive compounds. It is concluded that the production of P. setacea seedlings using G. albida is an alternative to offer phytomass to the herbal medicines industry based on passion fruit.

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Organic substrates are gaining popularity as a means of mitigating the negative effects of cadmium (Cd) stress on plant growth. The aim of the present study was to investigate the physio-biochemical attributes of Berberis integerrima bunge under Cd-contaminated soil. The pot experiment was carried out based on a completely randomized design (CRD) with six replicates. Cd stress was used as cadmium chloride (CdCl2) at 10, 20, and 30 mg Cd kg-1 dry soil. Biochar was applied at the doses of 125 g per pot, and vermicompost was used at the doses of 250 g per pot separately, and for their combination, they were used as 125 g per pot of BC + 250 g per pot of VC. The results showed higher Cd accumulation in both roots and leaves when the soil was polluted with Cd concentrations, but both BC and VC decreased the Cd accumulation in plant tissues. Although chlorophyll content and relative water content (RWC) decreased at 20 and 30 mg Cd kg-1 soil, BC and VC, particularly their combination, increased these traits. The highest total phenolic content (TPC) was observed in plants exposed to 20 mg Cd kg-1 soil and combined BC and VC. The total flavonoid content (TFC) was increased to 20 mg Cd kg-1 soil and then decreased to 30 mg Cd kg-1 soil. In addition, organic fertilizer promoted the plants' high accumulation of TFC. The greater activities of antioxidant enzymes including superoxide dismutase (SOD) and phenylalanine ammonia-lyase (PAL) were observed at 30 mg Cd kg-1 soil when organic substrates were added. The present study suggests the use of combined BC and VC lead to alleviate the adverse effects of Cd stress in B. integerrima.

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This study aimed to evaluate the effect of organic substrates on the growth yield, photosynthetic response, and nutritional profile of red leaf lettuce grown in different compositions of cocopeat (CP), sawdust (SD), and rice husk (RH). The result showed that the properties of substrates were influenced variably by their mixing ratios. The highest water holding capacity and moisture content were found in CP, and it provided the preferable pH, electrical conductivity, bulk density, and air-filled porosity in association with other categories of the substrate. Cocopeat-based media provides ample microclimate conditions in the root region of plants and increased their height, number of leaves, and fresh biomass components. The utmost dry biomass of plant parts also remarkably increased in CP; L*, a*, and b* chromaticity of leaves remained unchanged. The maximum chlorophyll content was attained in CP substrate, except for chlorophyll a/b, which was higher in RH. The net photosynthetic rate (PN), transpiration rate (E), and nitrate in leaves were enhanced substantially in CP, while it was lower in SD. Biochemical compositions and nutrients in leaves were likewise stimulated under the culture of cocopeat-based media. Results indicate that cocopeat, sawdust, and rice husk are a possible substrates mixture in a volume ratio of 3:1:1, which would be a better choice in the cultivation of red leaf lettuce.

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Organic amendments (OAs) application is a practical strategy to improve soil organic carbon (SOC) in agriculture. The present study evaluated the impact of different OAs on the transformation of carbon and the dynamics of microorganisms in a 77-day incubation experiment. The OA treatments applied included wheat straw (U + WS), pig manure (U + PM), compost (U + CP), and improved compost (U + IC), and the no amendment group was the CK. After incubation, the SOC increased significantly in the U + WS group, but the other OA treatments had no significant effect relative to the CK. Among the OA treatments, U + CP and U + IC had lower CO2-C cumulative mineralization and the highest humification of dissolved organic carbon (DOC). U + PM had the lowest SOC content and the lowest aromatization of DOC. Redundancy analyses (RDA) showed that the CO2-C cumulative mineralization directly influenced the DOC, extracted organic carbon (EOC) and microbial biomass carbon (MBC) in all treatments. Proteobacteria positively correlated with SOC and MBC, Bacteroidetes were significantly related to DOC, and Gemmatimonadetes had a significant negative relationship with CO2-C cumulative mineralization. These results showed that U + CP and U + IC were more conducive to carbon sequestration, and U + PM was the most unfavourable during the incubation. Wheat straw played an important role in the steady improvement of the SOC.

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This article contains the data of 11 organic substrates including physicochemical, biochemical and nutritional characterisations. Additionally, it includes for all substrates the data of organic matter fractionation into easily biodegradable, slowly biodegradable and inert fractions performed with anaerobic respirometry method. Finally, based on physicochemical characterisations and organic matter fractionation, a detailed methodology for the determination of input state variables required for the anaerobic digestion model N°1 (ADM1) was presented and the dataset for all substrates is provided. An example of calculation for one substrate illustrates the methodology for the determination of these variables. Data provided in this article could be useful to any person interested in modelling anaerobic digestion and particularly co-digestion. Data could be also used for implementation of a database for anaerobic digestion modelling.

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Seaweed (T1), sugarcane trash (T2), coir pith (T3) and vegetable waste (T4) with cowdung (1:1, w/w) were vermicomposted using Eudrilus eugeniae (50 days). The pH in vermicomposts showed a decrease while electrical conductivity showed increment. The organic matter content, organic carbon, lignin, cellulose, C/N and C/P ratios in vermicompost was significantly lower than compost. Total NPK contents of vermicompost were significantly elevated (P < 0.05) with 12.04-63.75%, 19.05-31.58% and 22.47-42.55%, respectively. The significantly higher growth rate of 1.41 and 7.74 mg/worm/day was observed in T1 on 10th and 50th day respectively, with 23.91 initial C/N ratio; while it was 0.85 and 4.81 mg/worm/day in T4 with 69.81 initial C/N ratio. A similar pattern was reflected in cocoon production, hatchling success and hatchling number/cocoon. Results revealed that vermicompost quality, worm growth, and reproduction depend on C/N ratio. The study suggests that amendment materials like cowdung are necessary to reduce C/N ratio for effective vermicomposting.

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A incubation experiment was conducted to investigate whether combined amendment of biochar (B) and compost (CP), mushroom residue (M) and corn straw (Y) could enhance biodegradation of polycyclic aromatic hydrocarbons (PAHs) in contaminated soils. After 77 days of incubation, both B + M and B + Y significantly (p < 0.01) increased removal rate of PAHs compared with amendment of biochar alone. However, B+CP resulted in a significant (p < 0.01) decreasing of PAHs removal. Compared with no biochar and no organic substrates addition (CK) and B, both B+M and B+Y significantly (p < 0.01) enhanced concentrations of dissolved organic carbon (DOC) and were favorable for the microbial growth reflected by microbial biomass carbons (MBC) and emission of carbon dioxide. Redundancy analysis (RDA) indicated that B + CP, B + M and B + Y separated the bacterial community compared with CK and B. However, the community composition structure in B + CP was different from that of B + M and B + Y. Moreover, the abundance of some PAHs degraders and PAH degradation genes predicted by PICRUSt software was promoted by B + M or B + Y, whereas that was inhibited under B + CP. The present study suggested that both B + M and B + Y could accelerate biodegradation of PAHs mainly through increasing the concentration of DOC and the abundances of microbial PAH degraders in soils.

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Microbial transformation of arsenic species and their interaction with the carbon cycle play a major role in the mobility of this toxic metalloid in the environment. The influence of simple or complex organic substrates on arsenic bio-oxidation was studied using two bacterial strains: one - the arsenivorans strain of Thiomonas delicata - is able to use AsIII as sole energy source; the other, Herminiimonas arsenicoxydans, is not. Experiments were performed at two AsIII concentrations (75 and 2 mg/L). At 75 mg/L As, for both strains, expression of aioA gene decreased when yeast extract concentration was raised from 0.2 to 1 g/L. At 2 mg/L As, the presence of either yeast extract or simple (succinate or acetate) organic substrates in the medium during bacterial growth decreased the AsIII-oxidation rate by both strains. When added specifically during oxidation test, yeast extract but not simple organic substrates seems to have a negative effect on AsIII oxidation. Taken together, results confirm the negative influence of simple or complex organic substrates on the kinetics of microbial AsIII oxidation and suggest that this effect results from different mechanisms depending on the type of organic substrate. Further, for the first time, the influence of a complex organic substrate, yeast extract, on aioA gene expression has been evidenced.

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In agriculture, the increasing scarcity of arable land and the increase in extreme weather conditions has led to a large proportion of crops, especially vegetables, being cultivated in protected soilless cultivation methods to provide people with sufficient and high-quality food. Rockwool has been used for decades as a soil substitute in soilless cultivation. Since rockwool is not biodegradable, it is disposed in landfills after its use, which nowadays leads to ecological concerns and drives the search for alternative substrates, especially organic materials. The objectives of this study were to investigate the effects of organic materials (wood chips, sphagnum moss, and hemp fibers) in relation to rockwool substrate on plant growth and quality of lettuce as a result of physical and chemical properties of the mentioned substrates. We were able to show that sphagnum moss is a suitable substitute substrate for lettuce cultivation, contrary to hemp. All investigated substrates presented good physical properties, but differed in their decomposition stability. Within 8 weeks, 30% of the hemp and about 10% of both sphagnum and wood materials were degraded. It was concluded that the increased microbiological activity immobilized nitrogen and led to oxygen deficiency in the rhizosphere and resulted in increased phenolic acid contents in lettuce but poor yield on hemp. Sphagnum caused a pH decrease and accumulation of ammonium in the nutrient solution and allowed the highest yield for lettuce at moderate phenolic acid contents. Low yields were obtained on wood, which could possibly be increased by optimized nutrient solution, so that wood as an alternative to rockwool was not excluded. By applying used organic substrates as soil additives on arable land, the nutrients accumulated in it might fertilize the open field crops, thus saving mineral fertilizers. This, together with the avoidance of waste, would contribute to a greater sustainability.

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Direct interspecies electron transfer (DIET) via electrically conductive pili (e-pili) and c-type cytochrome between acetogens and methanogens has been proposed as an essential pathway for methane production. Supplements of conductive materials have been extensively found to promote methane production in microbial anaerobic treatment systems. This review comprehensively presents recent findings of DIET and the addition of conductive materials for methanogenesis and summarizes important results through aspects of electron flux, organic degradation, and microbial interaction. Conductive materials improve DIET and methanogenesis by acting as either substitute of e-pili or electron conduit between e-pili and electron acceptors. Other effects of conductive materials such as the change of redox potential may also be important factors for the stimulation. The type and organic loading rate of substrates affect the occurrence of DIET and stimulating effects of conductive materials. Geobacter, which can participate in DIET, were less enriched in anaerobic systems cultivated with non-ethanol substrates, suggesting the existence of other syntrophs with the capability of DIET. The coupling of communication systems such as quorum sensing may be a good strategy to achieve the formation of biofilm or granule enriched with syntrophic partners capable of DIET.

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The effects of plant-based carbon source addition on wastewater NO3--N removal and the involved microorganisms, especially denitrifying bacteria, were investigated. A synthetic wastewater (NO3--N, 15 mg/L) was treated through the batch experiment, which included three inoculation cycles (7 days/cycle), and was conducted at 25 °C. Four natural plant substrates, namely, rice straw (RS), wheat straw (WS), ryegrass (RG), and reed (RD), were used as carbon sources and supplemented at the rate of 1% (w/v). The results showed that both RS and WS performed well in promoting NO3--N removal (79.55-97.07%). While RG removed only 22.08% of NO3--N in the first cycle, the removal efficiency increased afterward (86.09-95.82%). Conversely, the NO3--N removal rate of RD decreased from 95.10 to 24.77% as a result of its low ability to supply carbon. With respect to the microorganisms, the RS treatment resulted in more bacteria and denitrifying genes such as narG, nirK, nirS, and norB than other treatments, while the highest number of nosZ gene copies was recorded in the WS treatment. Sequencing results revealed that Firmicutes (18.19-56.96%), Proteobacteria (38.82-74.80%), and Bacteroidetes (3.15-4.15%) were three dominant bacterial phyla for RS, WS, and RD treatments. Furthermore, the genera Enterobacter, Massilia, and Bacillus were the main denitrifying bacteria participating in the NO3--N removal. Furthermore, correlation analysis indicated that the denitrifying genus Sphingobacterium played an important role in enhancing nitrogen removal. This study suggested that RS is the superior plant-based carbon source for denitrifying bioreactors used in agricultural runoff treatment.

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