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Bioplastics were first introduced as environmentally friendly materials, with properties similar to those of conventional plastics. A bioplastic is defined as biodegradable if it can be decomposed into carbon dioxide under aerobic degradation, or methane and CO2 under anaerobic conditions, inorganic compounds, and new cellular biomass, by the action of naturally occurring microorganisms. This definition however does not provide any information on the environmental conditions, timescale and extent at which decomposition processes should occur. With regard to the aquatic environment, recognized standards have been established to assess the ability of plastics to undergo biodegradation; however, these standards fail to provide clear targets to be met to allow labelling of a bioplastic as biodegradable. Moreover, these standards grant the user an extensive leeway in the choice of process parameters. For these reasons, the comparison of results deriving from different studies is challenging. The authors analysed and discussed the degree of biodegradability of a series of biodegradable bioplastics in aquatic environments (both fresh and salt water) using the results obtained in the laboratory and from on-site testing in the context of different research studies. Biochemical Oxygen Demand (BOD), CO2 evolution, surface erosion and weight loss were the main parameters used by researchers to describe the percentage of biodegradation. The results showed a large variability both in weight loss and BOD, even when evaluating the same type of bioplastics. This confirms the need for a reference range of values to be established with regard to parameters applied in defining the biodegradability of bioplastics.
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Biodegradación Ambiental , Contaminantes Químicos del Agua , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/metabolismo , Plásticos Biodegradables/metabolismo , Plásticos/metabolismo , Monitoreo del Ambiente/métodosRESUMEN
Recent studies have investigated the use of Black Soldier Fly (BSF) larvae as a promising biological treatment process for high organic content wastewater (i.a. Leachate from municipal solid waste landfill, food processing effluents), achieving both high treatment efficiency and production of secondary resources from larval biomass (i.a. Proteins and lipids). The present study was aimed at achieving a better understanding of how organic concentration and load might influence treatment performance. Larvae were fed with three artificial wastewaters characterised by same organic substances quality (degree of biodegradability and oxidation of the organic content measured respectively as BOD/COD and TOC/COD ratios) but different organic concentrations. Each type of wastewater was tested at four different loads. Treatment performance was assessed by monitoring both larval growth (in terms of weight variation, mortality and prepupation), and variation of wastewater quality and quantity to determine organic substrate consumption (measured in terms of Total Organic Carbon, TOC). Larval starvation was observed in all tests when TOC concentrations dropped below approx. 1000 mg C/L, which, for the tested wastewater, could be assumed as the limit value for adopting BSF larvae process. Substrate concentration in the feed (mgC/L) influenced larval growth (in terms of maximum wet weight, prepupation and mortality) only when organic load was above 10 mgC/larva: the higher the load, the higher the positive impact of the substrate concentration. On the contrary, the specific substrate consumption rate (vS, mgC/larva/day) appeared not to be influenced by substrate concentration but only by the organic load, with a Michaelis Menten like relationship. Accordingly, substrate load can be assumed as a design parameter for BSF treatment process, while substrate concentration might only influence potential resource recovery from larval biomass.
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Dípteros , Aguas Residuales , Animales , Larva/química , Larva/metabolismo , Alimentación Animal/análisis , Residuos SólidosRESUMEN
The use of Black Soldier Fly (BSF) larvae in the treatment of biowaste, including food waste, represents a promising new (waste) treatment option. In line with an increasing use of starch-based bioplastics in food packaging, (e.g. shopper films), food waste contamination by these polymers is expected to rise, but the fate of these materials and impact produced on the BSF treatment process remain to be clarified. In the present study, food waste contaminated by starch-based bioplastic film was treated using a BSF larvae process with the aim of investigating both the effect of bioplastics on process performance and the effect of BSF larvae on bioplastic degradation. Larvae treatment performance was assessed by monitoring substrate degradation process and larvae growth in terms of weight variation and development time. Bioplastic degradation (both in the larvae process and in a larvae-free control test) was assessed by means of visual inspection, Scanning Electron Microscopy (SEM), Fourier Transform InfraRed spectroscopy (FTIR), Differential Scanning Calorimetry (DSC) and ThermoGravimetric Analysis (TGA). The results obtained highlighted the absence of negative impacts of bioplastics on the BSF process, revealing a modestly higher degree of degradation in the larvae process compared to control test. The process however failed to achieve complete degradation of bioplastics, suggesting the need for additional post-processing treatments.
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Dípteros , Eliminación de Residuos , Animales , Larva/metabolismo , Alimentos , AlmidónRESUMEN
Sustainable landfill continues to play a fundamental role in closing the loop of residual materials of the circular economy. The sustainable landfill relies on both pretreatments and in situ treatments to stabilize the residual waste and immobilize the contaminants, achieving the final storage quality (FSQ) within one generation (typically 30 years). The aim of the study was to investigate the efficiency of the waste washing pretreatment in reducing the waste leaching fraction prior to landfilling, and in decreasing the time needed to reach the FSQ. A laboratory scale washing test was performed on three different kinds of residues from municipal solid waste treatment, usually landfilled: residues sieved from separately collected bio-waste (RB); residues sieved from compost (RC); and residues sieved from mixed waste treatment-plastic line (RP). Column landfill simulation tests were performed to predict and compare the landfill long-term emissions of both washed and raw residues. The results revealed that the washing pretreatment significantly reduced the leachable fraction of contaminants, decreasing the time needed to reach the chemical oxygen demand and ammonia FSQ limits. However, RP residue was the only one respecting the FSQ limits within 30 years.
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Eliminación de Residuos , Contaminantes Químicos del Agua , Eliminación de Residuos/métodos , Residuos Sólidos , Instalaciones de Eliminación de Residuos , Análisis de la Demanda Biológica de Oxígeno , Amoníaco , Contaminantes Químicos del Agua/análisisRESUMEN
Membrane filtration processes have been used to treat landfill leachate. On the other hand, closing the leachate treatment loop and finding a final destination for landfill leachate membrane concentrate (LLMC) - residual stream of membrane systems - is challenging for landfill operators. The re-introduction of LLMC into the landfill is typical; however, this approach is critical as concentrate pollutants may accumulate in the leachate treatment facility. From that, leachate concentrate management based on resource recovery rather than conventional treatment and disposal is recommended. This work comprehensively reviews the state-of-the-art of current research on LLMC management from leachate treatment plants towards a resource recovery approach. A general recovery train based on the main LLMC characteristics for implementing the best recovery scheme is presented in this context. LLMCs could be handled by producing clean water and add-value materials. This paper offers critical insights into LLMC management and highlights future research trends.
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Contaminantes Químicos del Agua , Instalaciones de Eliminación de ResiduosRESUMEN
The biological treatment process based on the metabolism of Black Soldier Fly (BSF) larvae proved to be a highly promising technique for the treatment of high organic content (HOC) wastewater, such as sewage from food industries, leachate from municipal solid waste (MSW) landfill, etc. The present study was aimed at achieving a better understanding of how biodegradability and degree of oxidation of organic content might influence treatment performance and biomass quality. Six leachates characterised by similar COD (Chemical Oxygen Demand) but different BOD5/COD (Biochemical Oxygen demand/COD) and TOC/COD (Total Organic Carbon/COD) ratios were tested. By combining these ratios, the BOD5/TOC ratio was introduced to take into account the effect of both leachate properties (biodegradability and oxidation degree). Process treatment performance was significantly influenced by the quality of organic substances. Higher BOD5/TOC values (higher biodegradability and lower oxidation degree) resulted in a greater and faster larvae growth, with final wet weight of between 49.2 and 91.9mg/larva; lower mortality between 5 and 32%; higher prepupation percentages ranging from 4 to 21% and higher specific substrate consumption rate with values varying from 0.051 to 0.063 mgTOC/mg larva/d, up to 3-fold higher than values obtained using conventional activated sludge based on COD consumption. Conversely, no significant differences were detected in larvae protein and lipid contents, including the profiling of fatty acids.
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Dípteros , Aguas Residuales , Animales , Larva/metabolismo , Oxidación-Reducción , Aguas del Alcantarillado/químicaRESUMEN
When approaching the study of new processes for leachate treatment, each influencing variable should be kept under control to better comprehend the treatment process. However, leachate quality is difficult to control as it varies dramatically from one landfill to another, and in line with landfill ageing. To overcome this problem, the present study investigated the option of preparing a reliable artificial leachate in terms of quality consistency and representativeness in simulating the composition of real municipal solid waste (MSW) leachate, in view of further investigate the recent treatment process using black soldier fly (BSF) larvae. Two recipes were used to simulate a real leachate (RL): one including chemical ingredients alone (artificial synthetic leachate-SL), and the other including chemicals mixed with artificial food waste (FW) eluate (artificial mixed leachate-ML). Research data were analysed, elaborated and discussed to assess simulation performance according to a series of parameters, such as Analytical representativeness, Treatment representativeness (in this case specific for the BSF larvae process), Recipe relevance, Repeatability and Flexibility in selectively modifying individual quality parameters. The best leachate simulation performance was achieved by the synthetic leachate, with concentration values generally ranging between 97% and 118% of the RL values. When feeding larvae with both RL and SL, similar mortality values and growth performance were observed.
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Dípteros , Eliminación de Residuos , Animales , Alimentos , Larva , Residuos Sólidos/análisisRESUMEN
Circular economy is considered a new chance to build a more sustainable world from both the social and the economic point of view. In this Essay, the possible contribution of inorganic chemistry towards a smooth transition to circularity in inorganic materials design and production is discussed by adopting an interdisciplinary approach.
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The benefits of using Black Soldier Fly (BSF) larvae in biowaste treatment include: commercial value of the stabilized residue, production of biomass rich in fats and proteins, suitable both for biodiesel production and animal feeding. The use of BSF for leachate treatment would introduce a blue low cost solution in the landfill technology, particularly appropriate in developing countries, where landfilling is still widely applied. This paper aimed to investigate the adaptability of BSF larvae to leachate environment, by using different leachate concentrations (25%, 50%, 75%, 100%) and two different feeding substrates: liquid (pure leachate) and semi-solid (wheat bran mixed with leachate). In all tests mortality was less than 50% and it was mainly linked to food shortages: the higher the nutrient content in leachate, the higher the larval development. Dry mass characterisation demonstrate that BSF prepupae biomass can be exploited as an alternative energy source in the production of biodiesel.
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Simuliidae , Alimentación Animal , Animales , Biocombustibles , Biomasa , LarvaRESUMEN
Semi-aerobic landfilling is applied increasingly as a sustainable technology worldwide, although frequently controversial results are achieved. The authors suggest that differences in water availability (climate, moisture content, etc.) and putrescible waste content are the key factors involved in controlling performance and efficiencies. The aim of the present study was to investigate the effect of inverse conditions (high/low) of these two factors. Six lab-scale lysimeters were specifically set up to correspond to three different conditions of water availability (wet conditions, dry conditions and artificially controlled watering under dry conditions) and two different waste types (high and low putrescible content). Lysimeters were operated for four months under thermal-insulated conditions and the quality and quantity of emissions monitored regularly. Concentrations of mobile ammonia and total organic carbon (TOC) in landfilled waste were modelled by means of first-order kinetics, and carbon and nitrogen mass balances were calculated. The best performance for the semi-aerobic process was achieved at a water availability of approximately 1.5-2.4 kgH2O/kgTS using the following two combinations: a) Waste with high putrescible content and no addition of external water due to the presence of sufficient endogenous water in the waste (moisture) to promote biological stabilisation of waste (Respiration index in 4 days, RI4 = 12.87 mgO2/gTS, BOD/COD < 0.05); b) Waste with low putrescible content and controlled watering (RI4 = 12.25 mgO2/gTS, BOD/COD < 0.04). The study highlighted how semi-aerobic landfilling operations should be carefully adjusted case by case according to waste quality and climate conditions.
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Eliminación de Residuos , Contaminantes Químicos del Agua , Carbono , Instalaciones de Eliminación de Residuos , AguaRESUMEN
In this study, the ability of H. illucens larvae (black soldier fly (BSF)) to metabolise different semisolid biowastes (e.g. kitchen waste, animal manure) has been applied to the treatment of landfill leachate. A testing programme has been developed by mixing leachate with three different solid supports: wheat bran, a biodegradable nutrient substrate, brewers' spent grain, a biodegradable nutrient residue from the brewery industry and sawdust, a low biodegradable residue from the wood industry. Larvae growth rate was monitored in terms of weight variation, mortality and time to reach the prepupal stage. Prepupal biomass composition was analysed in terms of crude protein, lipids and fatty acids. Substrates were monitored at the beginning and the end of tests for total solids (TS), total organic carbon (TOC), total Kjeldahl nitrogen (TKN), ammonia and (whenever significant) the 7-day Respirometric Index (RI7). The best performance was observed with wheat bran and brewers' spent grain, achieving an average larval weight ranging from 155.1 to 226.1 mg (w/w) with prepupation of more than 80% over 21 days. The initial TS, TOC and nitrogen content in feeding substrates had been metabolised (gasified and accumulated in prepupal biomass) by approximately 55%, 60% and 48%, respectively. Dry mass characterisation displayed a significant content of fats and proteins. The analysis demonstrated the suitability of BSF prepupal biomass for the production of biodiesel; however, the potential use of proteins as an animal feed needs further studies for assessing the presence of contaminants.
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Simuliidae , Alimentación Animal , Animales , Biocombustibles , Larva , EstiércolRESUMEN
This study compares the performance of a lab-scale pre-anoxic and post-aerobic submerged dynamic membrane bioreactor (DMBR) with similar studies on conventional membrane bioreactors (MBRs) for the treatment of old landfill leachate (LFL) while presenting a strategy to achieve stable DMBR operation. The results suggested that DMBR performed similar, or in some cases, better than MBRs. Like conventional MBRs treating LFL, DMBR can also accommodate large variations in operating parameters including influent feed composition and loading rates and thus, it can guarantee long term stable bioreactor operation (total nitrogen removal up to 98%) with acceptable effluent quality (Turbidityâ¯<â¯10 NTU). The results also demonstrated that gradual increment in influent LFL concentration was found to be effective for a stable DMBR operation however, it significantly deteriorated dynamic membrane (DM) filtration performance (pâ¯<â¯10E-7), resulting in higher fouling rate and deteriorated effluent quality. Nonetheless, poor DM performance and higher fouling rate were effectively controlled by using lower mesh porosity (52⯵m instead of 200⯵m) and increase in DM effective filtration area.
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Contaminantes Químicos del Agua , Reactores Biológicos , NitrógenoRESUMEN
Phytotreatment capping in closed landfills is a promising, cost-effective, in situ option for sustainable leachate treatment and might be synergistically coupled with energy crops to produce renewable energy (e.g.: biodiesel or bioethanol). This study proposes to use 0.30â¯m of soil as growing substrate for plants cultivated on the temporary cover of closed landfills. Once the leachate phytotreatment process is no longer required, 0.70â¯m of the same soil would be added to attain the final top cover configuration. This solution would entail saving the costs of excavation and backfilling. However, worsening of the initial soil quality due to potential contaminant transfer from the liquid to the solid matrix must be avoided because EU legislation (such as that in Italy) fixes concentration limits for contaminants in soil. In this research, samples of soil used as substrate in a lab-scale leachate phytotreatment test with sunflowers were analysed to provide chemical characterization before, during, and at the end of the experiment. The results showed that the phytotreatment activity did not increase initial contaminant concentrations. These results are reinforced by those from ecotoxicological bioassays in which Eisenia fetida (earthworms), Lepidium sativum (cress), Folsomia candida (collembola), and Caenorhabditis elegans and Steinernema carpocapsae (nematodes) were used. It was observed that, by the end of the experiment, the substrate soil did not affect the earthworms, collembola and nematode behaviour, or the growth of cress.
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Eliminación de Residuos , Contaminantes del Suelo , Contaminantes Químicos del Agua , Productos Agrícolas , Italia , Suelo , Instalaciones de Eliminación de ResiduosRESUMEN
This study investigated the behaviour of dynamic membrane (DM) filtration for the treatment of stabilised landfill leachate in a bench-scale pre-anoxic and aerobic submerged dynamic membrane bioreactor (DMBR). Four meshes with different openings (10, 52, 85 and 200⯵m) were tested to support the development of DM. Differences were observed among the meshes in supporting the development of the cake layer constituting the DM. The treatment of landfill leachate had an impact on sludge characteristics resulting in deteriorated filtration performance of the DM. Effluent turbidity was often higher than 100 NTU for larger mesh pore size (85 and 200⯵m). Low effluent turbidity was achieved with meshes with 10 and 52⯵m (13⯱â¯2 and 26⯱â¯4 NTU, respectively) although at membrane fluxes lower than 10â¯L m- 2 h-1. The bioreactor exhibited a moderate organics removal of 50-60% and an ammonia oxidation between 80 and 90%. Incomplete nitrification was observed due to increased concentrations of free ammonia and free nitrous acid, with nitrite effluent concentrations up to 1062 mgNO2--N L-1. Due to the large presence of refractory organic matter in landfill leachate, denitrification was limited resulting in a total nitrogen removal of approximately 20%.
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Desnitrificación , Contaminantes Químicos del Agua , Amoníaco , Reactores Biológicos , Nitrificación , NitrógenoRESUMEN
In this study, a top cover system is investigated as a control for emissions during the aftercare of new landfills and for old landfills where biogas energy production might not be profitable. Different materials were studied as landfill cover system in lab-scale columns: mechanical-biological pretreated municipal solid waste (MBP); mechanical-biological pretreated biowaste (PB); fine (PBSf) and coarse (PBSc) mechanical-biological pretreated mixtures of biowaste and sewage sludge, and natural soil (NS). The effectiveness of these materials in removing methane and sulphur compounds from a gas stream was tested, even coupled with activated carbon membranes. Concentrations of CO2, CH4, O2, N2, H2S and mercaptans were analysed at different depths along the columns. Methane degradation was assessed using mass balance and the results were expressed in terms of methane oxidation rate (MOR). The highest maximum and mean MOR were observed for MBP (17.2gCH4/m2/hr and 10.3gCH4/m2/hr, respectively). Similar values were obtained with PB and PBSc. The lowest values of MOR were obtained for NS (6.7gCH4/m2/hr) and PBSf (3.6gCH4/m2/hr), which may be due to their low organic content and void index, respectively. Activated membranes with high load capacity did not seem to have an influence on the methane oxidation process: MBP coupled with 220g/m2 and 360g/m2 membranes gave maximum MOR of 16.5gCH4/m2/hr and 17.4gCH4/m2/hr, respectively. Activated carbon membranes proved to be very effective on H2S adsorption. Furthermore, carbonyl sulphide, ethyl mercaptan and isopropyl mercaptan seemed to be easily absorbed by the filling materials.
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Contaminantes Atmosféricos/análisis , Metano/análisis , Eliminación de Residuos/métodos , Compuestos de Azufre/química , Instalaciones de Eliminación de Residuos , Contaminantes Atmosféricos/química , Metano/química , Oxidación-Reducción , Compuestos de Azufre/análisisRESUMEN
Efficient and economic reuse of waste is one of the pillars of modern environmental engineering. In the field of domestic sewage management, source separation of yellow (urine), brown (faecal matter) and grey waters aims to recover the organic substances concentrated in brown water, the nutrients (nitrogen and phosphorous) in the urine and to ensure an easier treatment and recycling of grey waters. With the objective of emphasizing the potential of recovery of resources from sewage management, a lab-scale research study was carried out at the University of Padova in order to evaluate the performances of oleaginous plants (suitable for biodiesel production) in the phytotreatment of source separated yellow and grey waters. The plant species used were Brassica napus (rapeseed), Glycine max (soybean) and Helianthus annuus (sunflower). Phytotreatment tests were carried out using 20L pots. Different testing runs were performed at an increasing nitrogen concentration in the feedstock. The results proved that oleaginous species can conveniently be used for the phytotreatment of grey and yellow waters from source separation of domestic sewage, displaying high removal efficiencies of nutrients and organic substances (nitrogen>80%; phosphorous >90%; COD nearly 90%). No inhibition was registered in the growth of plants irrigated with different mixtures of yellow and grey waters, where the characteristics of the two streams were reciprocally and beneficially integrated.
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Aguas del Alcantarillado , Eliminación de Residuos Líquidos/métodos , Biodegradación AmbientalRESUMEN
Dynamic membrane represents a cost effective alternative to conventional membranes by employing fouling as a means of solid-liquid separation. This study evaluated the effects of initial flux on both development rate of dynamic membrane and bioreactor performance during two consecutive experiments. The dynamic membrane was developed over a 200 µm mesh and the reactor was operated under anaerobic conditions. It was found that the effect of an initial higher applied flux on dynamic membrane development was more pronounced than mixed liquor suspended solid concentration inside the bioreactor. The development of the dynamic membrane was therefore positively associated with the applied flux. The rapid development of the dynamic membrane during the second experimental run at high initial fluxes and lower MLSS concentrations also affected the performance of the bioreactor in terms of more efficient COD removal and biogas production. A major shortcoming of applying higher initial applied flux was the formation of a denser and robust dynamic membrane layer that was resistant to applied hydraulic shear to control desired permeability and thus represented an obstacle in maintaining a long term operation with sustainable flux at lower transmembrane pressure (TMP).
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Biocombustibles , Reactores Biológicos , Membranas Artificiales , Eliminación de Residuos Líquidos/instrumentación , Anaerobiosis , Filtración , Permeabilidad , Eliminación de Residuos Líquidos/métodos , Aguas ResidualesRESUMEN
A lack of standardization in monitoring protocols has hindered the accurate evaluation of microplastic (MP) pollution in the open sea and its potential impacts. As sampling techniques significantly influence the amounts of MPs contained in the sample, the aim of this study was to compare two sampling methods: Manta trawl (size selective approach) and grab sampling (volume selective approach). Both approaches were applied in the open sea surface waters of the North-east Atlantic Ocean. Onshore sample processing was carried out using the innovative tape lifting technique, which affords a series of advantages, including prevention of airborne contamination during analysis, without compromising integrity of the results. The results obtained indicated an MP concentration over four orders of magnitude higher using grab sampling compared to the Manta net approach (mean values equal to 0.24 and 4050 items/m3, respectively). Consequently, the sole quantification of MPs using results obtained with the Manta trawl resulted in a marked underestimation of abundance. Nevertheless, the grab sampling technique is intricately linked to a risk of collecting non-representative water volumes, consequently leading to an overestimation of MPs abundance and a significant inter-sample variability. Moreover, the latter method is unsuitable for use in sampling larger MPs or in areas with low concentrations of MP pollution. The optimal sampling method therefore is dependent on the specific objectives of the study, often resulting in a combination of size and volume selective methods. The results of this study have the potential to contribute to the standardization of monitoring protocols for microplastics, both during the sampling phase and sample processing.
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System dynamics (SD) is widely recognized as a tool for simulating spatial and temporal dynamics in life cycle assessment (LCA) studies of the product system. However, there is no agreement on how SD and LCA could be applied effectively together in a consistent way. To address this gap, this research conducted a systematic literature review, analyzing 54 scientific articles published worldwide between 2010 and 2023, to explore the joint application of LCA with SD. The study aimed to answer three research questions: (1) What can be considered an integration of LCA and SD? (2) How can SD and LCA be effectively integrated? and (3)What are the advantages and constraints of this integration? The results highlighted the popularity of LCA and SD as impact assessment tools for sustainable design, each with its own strengths and limitations. Two primary integration types were identified when LCA was jointly applied with SD: (1) inclusion of the life cycle inventory and characterization factors in an SD model, and (2) inclusion of SD model results in an LCA. In the second type of integration, SD models the components of the technical system, and its outcomes served as input for scenario analysis, providing temporal and potentially spatial inventory data for the LCA model. The integrated approach offers a comprehensive understanding of product sustainability, aids decision-making, and enhances stakeholder engagement. The study also identifies knowledge gaps in the joint application of SD and LCA for environmental scenario analysis, suggesting the incorporation of optimization tools and strategy guidance for policy makers.
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Dark fermentation shares many features with anaerobic digestion with the exception that to maximize hydrogen production, methanogens and hydrogen-consuming bacteria should be inhibited. Heat treatment is widely applied as an inoculum pre-treatment due to its effectiveness in inhibiting methanogenic microflora but it may not exclusively select for hydrogen-producing bacteria. This work evaluated the effects of heat treatment on microbial viability and structure of anaerobic granular sludge. Heat treatment was carried out on granular sludge at 100 °C with four residence times (0.5, 1, 2 and 4 h). Hydrogen production of treated sludges was studied from glucose by means of batch test at different pH values. Results indicated that each heat treatment strongly influenced the granular sludge resulting in microbial communities having different hydrogen productions. The highest hydrogen yields (2.14 moles of hydrogen per mole of glucose) were obtained at pH 5.5 using the sludge treated for 4 h characterized by the lowest CFU concentration (2.3 × 10(3)CFU/g sludge). This study demonstrated that heat treatment should be carefully defined according to the structure of the sludge microbial community, allowing the selection of highly efficient hydrogen-producing microbes.