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Chitin was collected and extracted along different lifecycle stages of the Black Soldier Fly (BSF) (larvae, prepupae, pupae, flies, shedding & cocoons). The chitin content in the collected biomass ranged between 8% and 24%, with sheddings and cocoons being most rich in chitin. Purified chitin was subjected to a physicochemical evaluation based on FTIR, XRD, and TGA as well as a deacetylation step. The data indicated that BSF chitin was α-chitin with FTIR profiles matching closely to shrimp chitin and showing some differences compared to squid pen chitin (β-chitin). Small physicochemical differences were observed among the different BSF samples. Prepupae and cocoon chitin was more crystalline while chitin from larvae and sheddings had a lower thermal degradation temperature. In addition, sheddings were more difficult to purify. Further processing to chitosan showed that a deacetylation degree of 89% could be obtained for all samples after 3 h, although sheddings were found to be less reactive in the deacetylation process. Overall, the small differences in physicochemical properties that were detected between the BSF chitin samples did not prevent further processing of chitin to chitosan with the same degree of deacetylation via the same treatment.

Rhodomonas salina is a microalgal species, belonging to the cryptophytes, and is widely used as aquaculture feed because of its high nutritional profile and phycoerythrin content. This study investigated the effect of pH on the growth, biochemical composition, and taste of R. salina when cultivated on a semi-large scale under natural light conditions. Two tubular photobioreactors (200 L) were used for the cultivation of R. salina with sunlight as the only illumination source. Two different pH setpoints were applied, 7 and 8.5. Optimal temperature and nutrient conditions were applied, according to previous research findings. The results demonstrated that the productivity of R. salina was higher at pH 7, 0.06–0.14 gdry weight L−1 day−1, compared to pH 8.5, 0.03–0.12 gdry weight L−1 day−1. It was found that protein and total fatty acid concentrations were higher in the biomass that was produced at pH 8.5, 33.7% and 12.3% of dry weight, respectively, while at pH 7, the protein content was 31.9% and the total fatty acids 8.8% of dry weight. The phycoerythrin concentration, like protein, was higher at pH 8.5, 2.7% of dry weight, compared to pH 7, 1% of dry weight. The free amino acid and nucleotide profile of R. salina was affected by the pH, resulting in increased equivalent umami concentration at pH 7. For the sensory evaluation, an expert panel on algae flavors evaluated the effect of pH on the taste of R. salina, reporting that the biomass that was produced at pH 7 had more umami flavor than the biomass that was produced at pH 8.5, which was evaluated as more bitter.

This study aimed to map the nutritional profile and bioactivities of five microalgae that can be grown in Northwest Europe or areas with similar cultivation conditions. Next to the biochemical composition, the in vitro digestibility of carbohydrates, proteins, and lipids was studied for Chlamydomonas nivalis, Porphyridium purpureum, Chlorella vulgaris, Nannochloropsis gaditana, and Scenedesmus species biomass. These microalgae were also assessed for their ability to inhibit the angiotensin-1-converting enzyme (ACE-1, EC 3.4.15.1), which is known to play a role in the control of blood pressure in mammals. Large differences in organic matter solubility after digestion suggested that a cell disruption step is needed to unlock the majority of the nutrients from N. gaditana and Scenedesmus species biomass. Significant amounts of free glucose (16.4–25.5 g glucose/100 g dry algae) were detected after the digestion of C. nivalis, P. purpureum, and disrupted Scenedesmus. The fatty acid profiles showed major variations, with particularly high Ω-3 fatty acid levels found in N. gaditana (5.5 ± 0.5 g/100 g dry algae), while lipid digestibility ranged from 33.3 ± 6.5% (disrupted N. gaditana) to 67.1 ± 11.2% (P. purpureum). C. vulgaris and disrupted N. gaditana had the highest protein content (45–46% of dry matter), a nitrogen solubility after digestion of 65–71%, and the degree of protein hydrolysis was determined as 31% and 26%, respectively. Microalgae inhibited ACE-1 by 73.4–87.1% at physiologically relevant concentrations compared to a commercial control. These data can assist algae growers and processors in selecting the most suitable algae species for food or feed applications.

Insects are gaining interest as an alternative protein source for feed/food purposes. Although the lesser mealworm (LM) is commercially produced for human consumption, published data on its nutrient composition is scarce. This study reports on LM larvae reared on 18 different diets composed of side-streams to (1) determine the nutritional composition of the larvae and (2) study the effect of dietary changes on the larval nutrient composition. The LM larvae proved to be of good nutritional value with essential amino acids profiles comparable with that of beef and linoleic acid (C18:2) was the most dominant essential fatty acids in the larvae. The side-stream based diets varied on dry matter basis in protein (16&ndash, 34%) and lipid content (2&ndash, 19%). The nutrient content of the larvae reared on diets that supported good growth ranged between 37% and 49% of protein, 22% and 26% of lipid and 4% to 6% of chitin on dry matter basis. No significant correlations were identified between the larval protein or lipid content and that of the diet, but it was found between the diet nutrients and larval growth. Based on larval growth data and economic considerations, diets composed of wheat middlings with a 10&ndash, 15% inclusion of rapeseed meal were identified as suitable feed for LM. Highest larval yields were obtained with diets containing 15&ndash, 22% of proteins and 5&ndash, 10% of lipids.

Protein hydrolysates from lesser mealworm (Alphitobius diaperinus, LM) were obtained by enzymatic hydrolysis with protease from Bacillus licheniformis. A preliminary test performed for five hours of hydrolysis generated an insect protein hydrolysate with 15% of degree of hydrolysis (DH), optimum solubility property and oil holding capacity, but emulsifying and foaming ability were completely impaired. In order to investigate the potential implication of DH on techno-functional properties, a set of protein hydrolysates with a different DH was obtained by sub-sampling at different time points during three hours of enzymatic hydrolysis process. An increase in DH% had positive effects on the solubility property and oil holding ability, while a reduced emulsifying ability was observed up to five hours of hydrolysis. These results demonstrated that the enzymatic hydrolysis, if performed under controlled conditions and not for a long period, represents a valid method to extract high quality protein from insects with tailored techno-functionality, in order to produce tailored ingredients for feed and food purpose.

It is often impossible in practice to process micro-algae immediately after their cultivation and harvest. This study, therefore, aimed to identify appropriate storage conditions for the wet preservation of Porphyridium purpureum. Algae were stored either as a concentrate or as a dilute culture at 4 &deg, C, 8 &deg, C, or 20 &deg, C for 14 days and their quality was monitored. Concentrate storage tended to result in higher microbial numbers than dilute culture storage and clearly led to higher concentrations of malodorous organic acids. Butyric and isovaleric acid concentrations were about two orders of magnitude larger than their odor threshold values after 14 days of concentrate storage at 20 &deg, C. Average B-phycoeryhrin (B-PE) levels were slightly higher after concentrate storage (2.5 &plusmn, 0.2 g B-PE/100 g organic matter) than after dilute culture storage (2.2 &plusmn, 0.5 g B-PE/100 g organic matter), probably due to respiration losses of other organic compounds in the first case. Significant amounts of organic matter got lost during concentrate storage (4&ndash, 35%) as a result of carbohydrate degradation. The main restriction of concentrate storage was the rapid viscosity increase and formation of a weak gel structure complicating the later processing of the concentrates. These findings are highly relevant for P. purpureum cultivators and processors who have to store Porphyridium suspensions, even on a term of one day or less.

Algal lipids have gained wide interest in various applications ranging from biofuels to nutraceuticals. Given their complex nature composed of different lipid classes, a deep knowledge between extraction conditions and lipid characteristics is essential. In this paper, we investigated the influence of different pretreatments on lipid extraction with supercritical CO2 by a lipidomic approach. Pretreatment was found to double the total extraction yield, thereby reaching 23.1 wt.% comparable to the 26.9 wt.% obtained with chloroform/methanol. An increase in acylglycerides was concurrently observed, together with a nearly doubling of free fatty acids indicative of partial hydrolysis. Moreover, an alteration in the distribution of glyco- and phospholipids was noted, especially promoting digalactosyldiglycerides and phosphatidylcholine as compared to monogalactosyldiglycerides and phosphatidylglycerol. At optimized conditions, supercritical CO2 extraction provided a lipid extract richer in neutral lipids and poorer in phospholipids as compared to chloroform/methanol, though with a very similar fatty acid distribution within each lipid class.

The release of fine zerovalent iron (ZVI) particles in the environment after being introduced for in-situ treatment of compounds like chlorinated aliphatic hydrocarbons (CAHs) may raise questions toward environmental safety, especially for nanoscale materials. Classical single-species ecotoxicity tests do focus on aerobic conditions and are only relevant for the scenario when ZVI-particles reach surface water. Herein, we present an alternative approach where a CAH-degrading mixed bacterial culture was used as test-organisms relevant for the anaerobic subsurface. The impact of different ZVI particles on the bacterial culture was evaluated mainly by quantifying ATP, a reporter molecule giving a general indication of the microbial activity. These lab-scale batch tests were performed in liquid medium, without protecting and buffering aquifer material, as such representing worst-case scenario. The activity of the bacterial culture was negatively influenced by nanoscale zerovalent iron at doses as low as 0.05 g L−1. On the other hand, concentrations up to 2 g L−1 of several different types of microscale zerovalent iron (mZVI) particles stimulated the activity. However, very high doses of 15–30 g L−1 of mZVI showed an inhibiting effect on the bacterial community. Negative effects of ZVIs were confirmed by H2 accumulation in the batch reactors and the absence of lactate consumption. Observed inhibition also corresponded to a pH increase above 7.5, explicable by ZVI corrosion that was found to be dose-dependent. The obtained results suggest that low doses of mZVIs will not show severe inhibition effects on the microbial community once used for in-situ treatment of CAHs.

Microalgae harvesting is a major challenge because microalgal cells are small and carry a negative surface charge and biomass concentration in cultures is relatively low. The microalgal biomass (0.05%, w/w) needs to be concentrated to a paste with 15%–25% water content. This dewatering process is ideally performed in two stages, including a first preconcentration step in combination with a second dewatering step. Microalgae are a very heterogeneous group of organisms differing in size and shape and culture conditions. Applications of microalgal biomass range from low-value (biofuels) to high-value applications (nutraceuticals). It is therefore likely that the optimal harvesting technology differs between species, culture conditions, or the final application of the biomass. Harvesting should not cause contamination of the biomass or influence biomass quality. Finally, water recycling to reduce the water footprint is an important aspect to include into the harvesting process. This chapter gives an overview of several harvesting process options with the focus on their strengths and highlighting the aforementioned aspects. ispartof: Microalgae-based Biofuels and Bioproducts pages:113-132 ispartof: Woodhead Publishing Series in Energy pages:113-132 status: published

In this chapter, cell disruption techniques are reviewed as a function of the (future) industrial algae biorefinery. A variety of techniques and recent developments are discussed based on their effectiveness, product quality, and cost efficiency. Effectiveness (i.e, disruption efficiency in function of energy and time) unfortunately is often countered by poor product quality, and currently used industrial methods are generally energy intensive. Recent developments might tackle these issues and enhance the economics. Chemical techniques are often effective but can lead to impurities in the final product, which can be avoided by immobilizing chemical agents. Alternatively, several pressurized methods are promising because of high product quality and relatively low energy consumption. The latter can even be decreased significantly based on recent advances, which also counts for well-established techniques undergoing process optimization. The field, however, remains challenging to increase effectiveness and product quality to decrease downstream process steps and overall costs.

cis -1,2-Dichloroethene ( cis -DCE) and trichloroethene (TCE) are persistent, toxic and mobile pollutants in groundwater systems. They are both conducive to reductive dehalogenation and to oxidation by permanganate. In this study, the potential of dual element (C, Cl) compound specific isotope analyses (CSIA) for distinguishing between chemical oxidation and anaerobic reductive dechlorination of cis -DCE and TCE was investigated. Well-controlled cis -DCE degradation batch tests gave similar carbon isotope enrichment factors e C (‰), but starkly contrasting dual element isotope slopes Δ δ 13 C/Δ δ 37 Cl for permanganate oxidation (e C = − 26‰ ± 6‰, Δ δ 13 C/Δ δ 37 Cl ≈ − 125 ± 47) compared to reductive dechlorination (e C = − 18‰ ± 4‰, Δ δ 13 C/Δ δ 37 Cl ≈ 4.5 ± 3.4). The difference can be tracked down to distinctly different chlorine isotope fractionation: an inverse isotope effect during chemical oxidation (e Cl = + 0.2‰ ± 0.1‰) compared to a large normal isotope effect in reductive dechlorination (e Cl = − 3.3‰ ± 0.9‰) ( p ≪ 0.05). A similar trend was observed for TCE. The dual isotope approach was evaluated in the field before and up to 443 days after a pilot scale permanganate injection in the subsurface. Our study indicates, for the first time, the potential of the dual element isotope approach for distinguishing cis -DCE (and TCE) concentration drops caused by dilution, oxidation by permanganate and reductive dechlorination both at laboratory and field scale.

In this study we investigate the model uncertainties involved in predicting long-term permeable reactive barrier (PRB) remediation efficiency based on a lab-scale column experiment under accelerated flow conditions. A PRB consisting of 20% iron and 80% sand was simulated in a laboratory-scale column and contaminated groundwater was pumped into the column for approximately 1 year at an average groundwater velocity of 3.7E - 1 m d(-1). Dissolved contaminants (PCE. TCE, cis-DCE, trans-DCE and VC) and inorganic (Ca2+, Fe2+, TIC and pH) concentrations were measured in groundwater sampled at different times and at eight different distances along the column. These measurements were used to calibrate a multi-component reactive transport model, which subsequently provided predictions of long-term PRB efficiency under reduced flow conditions (i.e., groundwater velocity of 1.4E -3 m d(-1)), representative of a field site of interest in this study. Iron reactive surface reduction due to mineral precipitation and iron dissolution was simulated using four different models. All models were able to reasonably well reproduce the column experiment measurements, whereas the extrapolated long-term efficiency under different flow rates was significantly different between the different models. These results highlight significant model uncertainties associated with extrapolating long-term PRB performance based on lab-scale column experiments. These uncertainties should be accounted for at the PRB design phase, and may be reduced by independent experiments and field observations aimed at a better understanding of reactive surface deactivation mechanisms in iron PRBs. (C) 2012 Elsevier B.V. All rights reserved.

Microscale zerovalent iron (mZVI) reduces chlorinated aliphatic hydrocarbons (CAHs) to harmless compounds, but the sedimentation of the mZVI particles in the injection fluid limits the injectability of the particles during field applications. In this study, mZVI particles in suspension were stabilized by green polymer guar gum, which had a positive impact on mZVI stability, but decreased the reactivity of the particles towards CAHs by 1 to 8 times. Guar gum (GG) was found to adsorb onto the mZVI surface, inhibiting contact between the chlorinated compounds and the reactive iron surface. Indications were found for intermolecular hydrogen bonding between mZVI and the guar gum. Subsequent addition of commercially available enzymes resulted in the cleavage of the polysaccharide guar gum into lower molecular fragments, but not in improved reactivity. The reactivity recovery of guar gum coupled mZVI was recovered after intensive rinsing of the iron particles, removing the guar gum fragments from the particles. Overall, this study shows that CAHs can be treated efficiently by guar gum stabilized mZVI after reactivation by means of enzymatic breakdown and rinsing. (C) 2012 Elsevier B.V. All rights reserved.

The widespread use of methyl tertiary butyl ether (MTBE) as an additive in gasoline has resulted in serious soil and groundwater pollution. To tackle MTBE contamination in groundwater, bioremediation is considered a cost-effective and energy-efficient option. The design of an efficient, reliable and reproducible bioremediation strategy requires a thorough understanding of the (microbial) degradation kinetics of both MTBE and tertiary butyl alcohol (TBA), that is, the main intermediate during bioremediation of MTBE. To assist the design procedure, this research focuses on building a mathematical model to describe MTBE/TBA degradation kinetics by a bacterial consortium, that is, the VITO M-consortium. For the presented case study, the information obtained from experimental data is combined with knowledge regarding the biodegradation reactions in order to obtain a low-complexity, accurate mathematical model. The resulting model accurately describes MTBE/TBA degradation and has successfully been validated on ...

The injection of microscale zerovalent iron (mZVI) particles for groundwater remediation has received much interest in recent years. However, to date, monitoring of mZVI particle injection is based on chemical analysis of groundwater and soil samples and thus might be limited in its spatiotemporal resolution. To overcome this deficiency, in this study, we investigate the application of complex electrical conductivity imaging, a geophysical method, to monitor the high-pressure injection of mZVI in a field-scale application. The resulting electrical images revealed an increase in the induced electrical polarization (∼20%), upon delivery of ZVI into the targeted area, due to the accumulation of metallic surfaces at which the polarization takes place. Furthermore, larger changes (>50%) occurred in shallow sediments, a few meters away from the injection, suggesting the migration of particles through preferential flowpaths. Correlation of the electrical response and geochemical data, in particular the analysis of recovered cores from drilling after the injection, confirmed the migration of particles (and stabilizing solution) to shallow areas through fractures formed during the injection. Hence, our results demonstrate the suitability of the complex conductivity imaging method to monitor the transport of mZVI during subsurface amendment in quasi real-time.

International audience; We conducted a series of flow-through column experiments using aquifer sediment treated with zero valent iron (Fe-0) with or without active microbial sulfate reduction. The aim of this study was to investigate the stability of Zn removed by Fe-0 and impact of sulfate reducing bacteria (SRB). The leaching of Zn was assessed by flushing the aquifer in vertical glass columns with groundwater that was pre-treated with mixture of gases to simulate natural changes (pH and ORP) in subsurface environment (N-2 + CO2 for pH and N-2 + O-2 for ORP) along with a control (N-2 flushed). Zinc removed synergistically by Fe and SRBs was found to be more stable and did not leach from the aquifer sediment with pH or redox changes. We also observed that the Zn leaching in the aquifer treated only with Fe was more influenced by change in redox than by pH changes. These results were also confirmed by sequential metal extractions and scanning electron microscopy observations in the aquifer sediments. Mineral phases were predicted using geochemical modeling tool CHESS confirming that the metal removed in Fe-o + SRB conditions formed more stable precipitates and did not show any signs of leaching back to groundwater upon change in pH and redox conditions. (C) 2014 Elsevier B.V. All rights reserved.

In this study a numerical groundwater reactive transport model of a shallow groundwater aquifer contaminated with volatile organic compounds is developed. In addition to advective-dispersive transport, the model includes contaminant release from source areas, natural attenuation, abiotic degradation by a permeable reactive barrier at the site, and dilution by infiltrating rain. Aquifer heterogeneity is parameterized using pilot points for hydraulic conductivity, specific yield and groundwater recharge. A methodology is developed and applied to estimate the large number of parameters from the limited data at the field site (groundwater levels, groundwater concentrations of multiple chemical species, point-scale measurements of soil hydraulic conductivity, and lab-scale derived information on chemical and biochemical reactions). The proposed methodology relies on pilot point parameterization of hydraulic parameters and groundwater recharge, a regularization procedure to reconcile the large number of spatially distributed model parameters with the limited field data, a step-wise approach for integrating the different data sets into the model, and high performance computing. The methodology was proven to be effective in reproducing multiple contaminant plumes and in reducing the prior parameter uncertainty of hydraulic conductivity and groundwater recharge. Our results further indicate that contaminant transport predictions are strongly affected by the choice of the groundwater recharge model and flow parameters should be identified using both head and concentration measurements. (C) 2014 Elsevier B.V. All rights reserved.

An in situ mesocosm system was designed to monitor the in situ dynamics of the microbial community in polluted aquifers. The mesocosm system consists of a permeable membrane pocket filled with aquifer material and placed within a polypropylene holder, which is inserted below groundwater level in a monitoring well. After a specific time period, the microcosm is recovered from the well and its bacterial community is analyzed. Using this system, we examined the effect of benzene, toluene, ethylbenzene, and xylene (BTEX) contamination on the response of an aquifer bacterial community by denaturing gradient gel electrophoresis analysis of PCR-amplified 16S rRNA genes and PCR detection of BTEX degradation genes. Mesocosms were filled with nonsterile or sterile aquifer material derived from an uncontaminated area and positioned in a well located in either the uncontaminated area or a nearby contaminated area. In the contaminated area, the bacterial community in the microcosms rapidly evolved into a stable community identical to that in the adjacent aquifer but different from that in the uncontaminated area. At the contaminated location, bacteria with tmoA - and xylM/xylE1 -like BTEX catabolic genotypes colonized the aquifer, while at the uncontaminated location only tmoA -like genotypes were detected. The communities in the mesocosms and in the aquifer adjacent to the wells in the contaminated area consisted mainly of Proteobacteria . At the uncontaminated location, Actinobacteria and Proteobacteria were found. Our results indicate that communities with long-term stability in their structures follow the contamination plume and rapidly colonize downstream areas upon contamination.

The Sphingomonas genus hosts many interesting pollutant-degrading strains. Sphingomonas sp. EPA505 is the best studied polycyclic aromatic hydrocarbon (PAH)-degrading Sphingomonas strain. Based on 16S rRNA gene sequence analysis, Sphingomonas sp. strain EPA505 forms a separate branch in the Sphingomonas phylogenetic tree grouping exclusively PAH-degrading isolates. For specific PCR detection and monitoring of Sphingomonas sp. EPA505 and related strains in PAH-contaminated soils, a new 16S rRNA gene-based primer set was designed. The new primer set was shown to be highly selective for Sphingomonas sp. strain EPA505 as it only amplified DNA from strain EPA505 and not from other tested Sphingomonas strains or soil bacteria not belonging to the Sphingomonas genus. Using DNA extracts of a variety of inoculated PAH-contaminated soils, the primer pair was able to detect EPA505 in concentrations as low as 10(2) cells per gram of soil. Applying the new primer set, 16S rRNA gene fragments which were 99-100% similar to the corresponding gene of strain EPA505 were amplified from four of five PAH-contaminated soils. On the other hand, no PCR products were obtained from any of five tested uncontaminated soils. The preferential presence of EPA505 related Sphingomonas strains in PAH-contaminated soils with very different contamination profiles and different origin suggests an important role of this type of Sphingomonas in the natural Sphingomonas community colonizing PAH-contaminated sites.

Fast-growing mycobacteria are considered essential members of the polycyclic aromatic hydrocarbons (PAH) degrading bacterial community in PAH-contaminated soils. To study the natural role and diversity of the Mycobacterium community in contaminated soils, a culture-independent fingerprinting method based on PCR combined with denaturing gradient gel electrophoresis (DGGE) was developed. New PCR primers were selected which specifically targeted the 16S rRNA genes of fast-growing mycobacteria, and single-band DGGE profiles of amplicons were obtained for most Mycobacterium strains tested. Strains belonging to the same species revealed identical DGGE fingerprints, and in most cases, but not all, these fingerprints were typical for one species, allowing partial differentiation between species in a Mycobacterium community. Mycobacterium strains inoculated in soil were detected with a detection limit of 10(6) CFU g(-1) of soil using the new primer set as such, or approximately 10(2) CFU g(-1) in a nested PCR approach combining eubacterial and the Mycobacterium specific primers. Using the PCR-DGGE method, different species could be individually recognized in a mixed Mycobacterium community. This approach was used to rapidly assess the Mycobacterium community structure of several PAH-contaminated soils of diverse origin with different overall contamination profiles, pollution concentrations and chemical-physical soil characteristics. In the non-contaminated soil, most of the recovered 16S rRNA gene sequence did not match with previous described PAH-degrading Mycobacterium strains. In most PAH-contaminated soils, mycobacteria were detected which were closely related to fast-growing species such as Mycobacterium frederiksbergense and Mycobacterium austroafricanum, species that are known to include strains with PAH-degrading capacities. Interestingly, 16S rRNA genes related to M. tusciae sequences, a Mycobacterium species so far not reported in relation to biodegradation of PAHs, were detected in all contaminated soils. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

Chlorinated aliphatic hydrocarbons are common groundwater contaminants. One possible remediation option is in-situ reductive dechlorination by zero-valent iron, either by direct injection or as reactive barriers. Chlorinated ethenes (tetrachloroethene: PCE; trichloroethene: TCE) have received extensive attention in this context. However, another common groundwater pollutant, 1,1,1-trichlorethane (TCA), has attracted much less attention. We studied TCA reduction by three types of granular zero-valent irons in a series of batch experiments using polluted groundwater, with and without added aquifer material. Two types of iron were able to reduce TCA completely with no daughter product concentration increases (1,1-dichloroethane: DCA; chloroethane: CA). One type of iron showed slower reduction, with intermediate rise of DCA and CA concentrations. When evaluating the formation of daughter products, the tests on the groundwater alone showed different results than the groundwater plus aquifer batches: DCA did not temporarily accumulate in the batches with added aquifer material, contrary to the batches without added aquifer material. 1,1-dichloroethene (DCE, also present in the groundwater as an abiotic degradation product of TCA) was also reduced slower in the batches without added aquifer material than in the batches with aquifer material. Redox potentials gradually decreased to low values in batches with aquifer material without iron, while the batches with groundwater alone maintained a constant higher redox potential. Either adsorption processes or microbiological activity in the samples could explain these phenomena. Polymerase Chain Reaction (PCR: a targeted gene probe technique) for chlorinated aliphatic compound (CAH)-degrading bacteria confirmed the presence of Dehalococcoides sp. (chloroethene-degraders) but was negative for Desulfobacterium autotrophicum (a known co-metabolic TCA degrader). DCA reduction was rate determining: first-order half-lives of 300-350 h were observed. TCA was fully removed within hours. CA is resistant to reduction by zero-valent iron but it is known to hydrolyze easily. Since CA did not accumulate in our batches, it may have disappeared by the latter mechanism or it may not have formed as a major daughter product.

Bacterial strains of the genus Sphingomonas are often isolated from contaminated soils for their ability to use polycyclic aromatic hydrocarbons (PAH) as the sole source of carbon and energy. The direct detection of Sphingomonas strains in contaminated soils, either indigenous or inoculated, is, as such, of interest for bioremediation purposes. In this study, a culture-independent PCR-based detection method using specific primers targeting the Sphingomonas 16S rRNA gene combined with denaturing gradient gel electrophoresis (DGGE) was developed to assess Sphingomonas diversity in PAH-contaminated soils. PCR using the new primer pair on a set of template DNAs of different bacterial genera showed that the method was selective for bacteria belonging to the family Sphingomonadaceae . Single-band DGGE profiles were obtained for most Sphingomonas strains tested. Strains belonging to the same species had identical DGGE fingerprints, and in most cases, these fingerprints were typical for one species. Inoculated strains could be detected at a cell concentration of 10 4 CFU g of soil −1 . The analysis of Sphingomonas population structures of several PAH-contaminated soils by the new PCR-DGGE method revealed that soils containing the highest phenanthrene concentrations showed the lowest Sphingomonas diversity. Sequence analysis of cloned PCR products amplified from soil DNA revealed new 16S rRNA gene Sphingomonas sequences significantly different from sequences from known cultivated isolates (i.e., sequences from environmental clones grouped phylogenetically with other environmental clone sequences available on the web and that possibly originated from several potential new species). In conclusion, the newly designed Sphingomonas -specific PCR-DGGE detection technique successfully analyzed the Sphingomonas communities from polluted soils at the species level and revealed different Sphingomonas members not previously detected by culture-dependent detection techniques.

The paper reports a pilot injection test of microsized zerovalent iron (mZVI) dispersed in a guar gum shear thinning solution. The test was performed in the framework of the EU research project AQUAREHAB in a site in Belgium contaminated by chlorinated aliphatic hydrocarbons (CAHs). The field application was aimed to overcome those critical aspects which hinder mZVI field injection, mainly due to the colloidal instability of ZVI-based suspensions. The iron slurry properties (iron particles size and concentration, polymeric stabilizer type and concentration, slurry viscosity) were designed in the laboratory based on several tests (reactivity tests towards contaminants, sedimentation tests and rheological measurements). The particles were delivered into the aquifer through an injection well specifically designed for controlled-pressure delivery (approximately 10 bars). The well characteristics and the critical pressure of the aquifer (i.e. the injection pressure above which fracturing occurs) were assessed via two innovative injection step rate tests, one performed with water and the other one with guar gum. Based on laboratory and field preliminary tests, a flow regime at the threshold between permeation and preferential flow was selected for mZVI delivery, as a compromise between the desired homogeneous distribution of the mZVI around the injection point (ensured by permeation flow) and the fast and effective injection of the slurry (guaranteed by high discharge rates and injection pressure, resulting in the generation of preferential flow paths). A monitoring setup was designed and installed for the real-time monitoring of relevant parameters during injection, and for a fast determination of the spatial mZVI distribution after injection via non-invasive magnetic susceptibility measurements.

A pilot injection test with guar gum stabilized microscale zerovalent iron (mZVI) particles was performed at test site V (Belgium) where different chlorinated aliphatic hydrocarbons (CAHs) were present as pollutants in the subsurface. One hundred kilograms of 56 pm-diameter mZVI (similar to 70 g L-1) was suspended in 1.5 m(3) of guar gum (similar to 7 g L-1) solution and injected into the test area. In order to deliver the guar gum stabilized mZVI slurry, one direct push bottom-up injection (Geoprobe) was performed with injections at 5 depths between 10.5 and 8.5 m bgs. The direct push technique was preferred above others (e.g. injection at low flow rate via screened wells) because of the limited hydraulic conductivity of the aquifer, and to the large size of the mZVI particles. A final heterogeneous distribution of the mZVI in the porous medium was observed explicable by preferential flow paths created during the high pressure injection. The maximum observed delivery distance was 2.5 m. A significant decrease in 1,1,1-TCA concentrations was observed in close vicinity of spots where the highest concentration of mZVI was observed. Carbon stable isotope analysis (CSIA) yielded information on the success of the abiotic degradation of 1,1,1-TCA and indicated a heterogeneous spatio-temporal pattern of degradation. Finally, the obtained results show that mZVI slurries stabilized by guar gum can be prepared at pilot scale and directly injected into low permeable aquifers, indicating a significant removal of 1,1,1-TCA. (C) 2014 Elsevier B.V. All rights reserved.

Zerovalent iron (ZVI) abiotically degrades several chlorinated aliphatic hydrocarbons (CAHs) via reductive dechlorination, which offers perspectives for in situ groundwater remediation applications. The difference in reactivity between ZVI particles is often linked with their specific surface area. However, other parameters may influence the reactivity as well. Earlier, we reported for a set of microscale zerovalent iron (mZVI) particles the disappearance kinetic of different CAHs which were collected under consistent experimental conditions. In the present study, these kinetic data were correlated with the carbon, oxygen and sulfur content of mZVI particles. It was confirmed that not only the specific surface area affects the disappearance kinetic of CAHs, but also the chemical composition of the mZVI particles. The chemical composition, in addition, influences CAHs removal mechanism inducing sorption onto mZVI particles instead of dechlorination. Generally, high disappearance kinetic of CAHs was observed for particles containing less oxygen. A high carbon content, on the other hand, induced nonreactive sorption of the contaminants on the mZVI particles. To obtain efficient remediation of CAHs by mZVI particles, this study suggested that the carbon and oxygen content should not exceed 0.5% and 1% respectively. Finally, the efficiency of the mZVI particles may be improved to some extent by enriching them with sulfur. However, the impact of sulfur content on the reactivity of mZVI particles is less pronounced than that of the carbon and oxygen content. 2013 Elsevier Ltd. All rights reserved.

In the present study, controlled laboratory column experiments were conducted to understand the biogeochemical changes during the microbial sulfate reduction. Sulfur and oxygen isotopes of sulfate were followed during sulfate reduction in zero valent iron incubated flow through columns at a constant temperature of 20 +/- 1 degrees C for 90 d. Sulfur isotope signatures show considerable variation during biological sulfate reduction in our columns in comparison to abiotic columns where no changes were observed. The magnitude of the enrichment in delta S-34 values ranged from 9.4%(0) to 10.3%(0) compared to initial value of 2.3%(0), having total fractionation SS between biotic and abiotic columns as much as 6.1%(0). Sulfur isotope fractionation was directly proportional to the sulfate reduction rates in the columns. Oxygen isotopes in this experiment seem less sensitive to microbial activities and more likely to be influenced by isotopic exchange with ambient water. A linear relationship is observed between delta S-34 and delta O-18 in biotic conditions and we also highlight a good relationship between delta S-34 and sulfate reduction rate in biotic columns. (C) 2012 Elsevier Ltd. All rights reserved.

An important issue of concern for permeable reactive iron barriers is the long-term efficiency of the barriers due to the long operational periods required. Mineral precipitation resulting from the anaerobic corrosion of the iron filings and bacteria present in the barrier may play an important role in the long-term performance. An integrated study was performed on the Vapokon permeable reactive barrier (PRB) in Denmark by groundwater and iron core sample characterization. The detailed field groundwater sampling carried out from more than 75 well screens up and downstream the barrier showed a very efficient removal (>99%) for the most important CAHs (PCE, TCE and 1,1,1-TCA). However, significant formation of cis-DCE within the PRB resulted in an overall insufficient efficiency for cis-DCE removal. The detailed analysis of the upstream groundwater revealed a very heterogeneous spatial distribution of contaminant loading into the PRB, which resulted in that only about a quarter of the barrier system is treating significant loads of CAHs. Laboratory batch experiments using contaminated groundwater from the site and iron material from the core samples revealed that the aged iron material performed equally well as virgin granular iron of the same type based on determined degradation rates despite that parts of the cored iron material were covered by mineral precipitates (especially iron sulfides, carbonate green rust and aragonite). The PCR analysis performed on the iron core samples indicated the presence of a microbial consortium in the barrier. A wide range of species were identified including sulfate and iron reducing bacteria, together with Dehalococcoides and Desulfuromonas species indicating microbial reductive dehalogenation potential. The microbes had a profound effect on the performance of the barrier, as indicated by significant degradation of dichloromethane (which is typically unaffected by zero valent iron) within the barrier.

Eleven soil samples (contaminated and non-contaminated top soils and aquifers) from seven different locations in Belgium were examined in lab-scale batch microcosms simulating in situ conditions for their indigenous capacity to biodegrade methyl tert-butyl ether (MTBE). The effect of implementing nutrients or additional oxygen and of the presence of co-contaminants on MTBE degradation was investigated. All soils showed rapid degradation of benzene. On the other hand, only one site, historically contaminated with oxygenated fuel, provided soil samples showing relatively fast MTBE biodegradation. These soil samples originated from four different depths from the vadose and saturated zone. MTBE degradation kinetics differed between the samples of the saturated and non-saturated zone and depended on the implemented conditions. MTBE-biodegradation in the samples from the non-saturated zone started after a very short lag-phase (7 days), while long lag-phases (up to 270 days) were obtained with the other samples. Addition of extra nutrients stimulated MTBE degradation kinetics in microcosms containing the saturated soil samples while inhibiting effects were seen in the case of non-saturated soil samples. In contrast, implementing dissolved oxygen concentrations of 9.5 and 11.5 mg l(-1) led to lower degradation kinetics compared to 8 mg l(-1) in microcosms containing saturated soil samples, while stimulating effects were seen with the non-saturated soil samples. Addition of an extra carbon source like benzene or propane did increase in general the MTBE first order degradation rate constant. Differences in the eubacterial community composition between these depth samples were confirmed based on denaturing gradient gel electrophoresis (DGGE) patterns of PCR-amplified 16S rRNA gene fragments. The results of the presented study indicate that an aerobic MTBE biodegradation potential is not omnipresent in Belgian sub-soils.

Recent reports suggest that Mycobacterium is better adapted to soils containing poorly bioavailable polycyclic aromatic hydrocarbons (PAHs) compared to Sphingomonas. To study this hypothesis, artificial conditions regarding PAH profile and PAH bioavailability were induced in two PAH-contaminated soils and the response of the eubacterial, Mycobacterium, and Sphingomonas communities to these changed conditions was monitored during laboratory incubation. Soil K3663 with a relatively high proportion of high molecular weight PAHs was amended with phenanthrene or pyrene to artificially change the soil into a soil with a relatively increased bioavailable PAH contamination. Soil AndE with a relatively high proportion of bioavailable low molecular weight PAHs was treated by a single-step Tenax extraction to remove the largest part of the easily bioavailable PAH contamination. In soil K3663, the added phenanthrene or pyrene compounds were rapidly degraded, concomitant with a significant increase in the number of phenanthrene and pyrene degraders, and minor and no changes in the Mycobacterium community and Sphingomonas community, respectively. However, a transient change in the eubacterial community related to the proliferation of several γ-proteobacteria was noted in the phenanthrene-amended soil. In the extracted AndE soil, the Sphingomonas community initially developed into a more diverse community but finally decreased in size below the detection limit. Mycobacterium in that soil never increased to a detectable size, while the eubacterial community became dominated by a γ-proteobacterial population. The results suggest that the relative bioavailability of PAH contamination in soil affects bacterial community structure but that the behavior of Mycobacterium and Sphingomonas in soil is more complex than prospected from studies on their ecology and physiology.

tmoA and related genes encode the alpha-subunit of the hydroxylase component of the major group (subgroup 1 of subfamily 2) of bacterial multicomponent mono-oxygenase enzyme complexes involved in aerobic benzene, toluene, ethylbenzene and xylene (BTEX) degradation. A PCR-denaturing gradient gel electrophoresis (DGGE) method was developed to assess the diversity of tmoA-like gene sequences in environmental samples using a newly designed moderately degenerate primer set suitable for that purpose. In 35 BTEX-degrading bacterial strains isolated from a hydrocarbon polluted aquifer, tmoA-like genes were only detected in two o-xylene degraders and were identical to the touA gene of Pseudomonas stutzeri OX1. The diversity of tmoA-like genes was examined in DNA extracts from contaminated and non-contaminated subsurface samples at a site containing a BTEX-contaminated groundwater plume. Differences in DGGE patterns were observed between strongly contaminated, less contaminated and non-contaminated samples and between different depths, suggesting that the diversity of tmoA-like genes was determined by environmental conditions including the contamination level. Phylogenetic analysis of the protein sequences deduced from the amplified amplicons showed that the diversity of TmoA-analogues in the environment is larger than suggested from described TmoA-analogues from cultured isolates, which was translated in the DGGE patterns. Although different positions on the DGGE gel can correspond to closely related TmoA-proteins, relationships could be noticed between the position of tmoA-like amplicons in the DGGE profile and the phylogenetic position of the deduced protein sequence.

Mycobacterium is often isolated from polycyclic aromatic hydrocarbon (PAH)-contaminated soil as degraders of PAHs. In model systems, Mycobacterium shows attachment to the PAH substrate source, which is considered to be a particular adaptation to low bioavailability as it results into increased substrate flux to the degraders. To examine whether PAH-degrading Mycobacterium in real PAH-contaminated soils, in analogy with model systems, are preferentially associated with PAH-enriched soil particles, the distribution of PAHs, of the PAH-mineralizing capacity and of Mycobacterium over different fractions of a soil with an aged PAH contamination was investigated. The clay fraction contained the majority of the PAHs and showed immediate pyrene- and phenanthrene-mineralizing activity upon addition of 14C-labelled pyrene or phenanthrene. In contrast, the sand and silt fractions showed a lag time of 15-26 h for phenanthrene and 3-6 days for pyrene mineralization. The maximum pyrene and phenanthrene mineralization rates of the clay fraction expressed per gram fraction were three to six times higher than those of the sand and silt fractions. Most-probable-number (MPN)-polymerase chain reaction demonstrated that Mycobacterium represented about 10% of the eubacteria in the clay fraction, while this was only about 0.1% in the sand and silt fractions, indicating accumulation of Mycobacterium in the PAH-enriched clay fraction. The Mycobacterium community composition in the clay fraction represented all dominant Mycobacterium populations of the bulk soil and included especially species related to Mycobacterium pyrenivorans, which was also recovered as one of the dominant species in the eubacterial communities of the bulk soil and the clay fraction. Moreover, Mycobacterium could be identified among the major culturable PAH-degrading populations in both the bulk soil and the clay fraction. The results demonstrate that PAH-degrading mycobacteria are mainly associated with the PAH-enriched clay fraction of the examined PAH-contaminated soil and hence, that also in the environmental setting of a PAH-contaminated soil, Mycobacterium might experience advantages connected to substrate source attachment.

The metabolic pathway of the PAH fluorene and the cometabolic pathway of the PAHs phenanthrene, fluoranthene, anthracene and dibenzothiophene in Sphingomonas sp. LB 126 were examined. To our knowledge this is the first study on the cometabolic degradation of the three-ring PAHs phenanthrene, anthracene and the four-ring PAH fluoranthene by a fluorene-utilizing species. Metabolism of fluorene was shown to proceed via the 9-fluorenone pathway to form o-phthalic acid and protocatechuic acid. The cometabolic mono-hydroxylation found for phenanthrene, fluoranthene and anthracene shows similarity with the hydroxylation of fluorene. Several mono- and dihydroxy products and ring-cleavage products were identified for phenanthrene, fluoranthene and anthracene. It appeared that the cometabolism of those three compounds is a non-specific process, in contrast to the metabolism of fluorene. For dibenzothiophene the metabolites dibenzothiophene-5-oxide and dibenzothiophene-5,5 -dioxide were identified; these compounds appeared to be the products of a dead-end pathway. Since apart from dibenzothiophene no metabolites were found in very high concentrations for any of the other substrates, complete degradation is suggested, even for the cometabolic degradation of phenanthrene, fluoranthene and anthracene. (C) 2003 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

The promoter probe mini-Tn5-luxAB-tet was used to create a luxAB transcriptional fusion responding to fluorene in the fluorene utilising bacterium Sphingomonas sp. LB126. The mutant strain, named L-132, was impaired in fluorene utilisation and strongly emitted light upon addition of fluorene to the growth medium. L-132 was initially characterised and examined for its potential use as a whole-cell biosensor in the perspective of quantifying fluorene in environmental samples. Activity of the reporter gene as a response to fluorene was detectable after 30 rain and was optimal after 4 h. A linear response to fluorene concentrations within the water solubility range was achieved, with a detection limit of 200 mug per litre. Besides fluorene, L-132 weakly responded to the polycyclic aromatic hydrocarbons phenanthrene and dibenzothiophene, whereas strong responses were obtained with 9-fluorenone, 9-hydroxyfluorene, phthalic acid and protocatechuic acid. The latter four compounds are metabolites formed in course of fluorene degradation, which suggested that a fluorene metabolite rather than fluorene itself was the true inducer of the luxAB fusion in L-132. (C) 2001 Editions scientifiques et medicales Elsevier SAS.

Sphingomonas sp. LB126 is able to utilize fluorene as sole source of carbon and energy. In the present study, a mutagenic vector was constructed and a "plasmid rescue" strategy was set up to isolate a 16.5-kb DNA fragment containing genes required for fluorene degradation. A 14.5-kb portion of the cloned DNA was sequenced revealing thirteen open reading frames. Two encoded hypothetical proteins (FldE and FldY) similar to transcriptional regulators and one (ORF360) located on an IS-like element (ISSsp126) encoded a putative transposase. Three other putative proteins (FldB, FldU and FldV) displayed strong similarity with enzymes of the protocatechuate 4,5-degradation pathway utilized by Sphingomonaspaucimobilis SYK-6 for the degradation of lignin breakdown products. The remaining hypothetical proteins displayed only limited similarity with enzyme sequences available from databases. Suicide plasmid-directed mutagenesis and genetic complementations showed that integrity of the protocatechuate catabolic pathway was an absolute requirement for fluorene degradation to proceed. These findings were further supported by the analysis of metabolites in bacterial culture supernatants obtained from appropriate mutants. The results presented here demonstrated the suitability of the genetic tool constructed and supplied the first genetic evidence for the participation of a protocatechuate 4,5-degradation pathway in a bacterial fluorene degradation pathway.

Two different procedures were compared to isolate polycyclic aromatic hydrocarbon (PAH)-utilizing bacteria from PAH-contaminated soil and sludge samples, i.e., (i) shaken enrichment cultures in liquid mineral medium in which PAHs were supplied as crystals and (ii) a new method in which PAH degraders were enriched on and recovered from hydrophobic membranes containing sorbed PAHs. Both techniques were successful, but selected from the same source different bacterial strains able to grow on PAHs as the sole source of carbon and energy. The liquid enrichment mainly selected for Sphingomonas spp., whereas the membrane method exclusively led to the selection of Mycobacterium spp. Furthermore, in separate membrane enrichment set-ups with different membrane types, three repetitive extragenic palindromic PCR-related Mycobacterium strains were recovered. The new Mycobacterium isolates were strongly hydrophobic and displayed the capacity to adhere strongly to different surfaces. One strain, Mycobacterium sp. LB501T, displayed an unusual combination of high adhesion efficiency and an extremely high negative charge. This strain may represent a new bacterial species as suggested by 16S rRNA gene sequence analysis. These results indicate that the provision of hydrophobic sorbents containing sorbed PAHs in the enrichment procedure discriminated in favor of certain bacterial characteristics. The new isolation method is appropriate to select for adherent PAH-degrading bacteria, which might be useful to biodegrade sorbed PAHs in soils and sludge.

This study focusses on the fractionation of proteins and lipids from insect biomass, more specifically from black soldier fly larvae. Insects are rich in proteins and lipids and offer potential as a protein source for food and feed. Fractionation processes often focus on extracting only one compound (lipid or protein), neglecting the impact of the extraction method on other compounds. The current study aimed at the development of a robust fractionation process for wet insect biomass, targeting enriched protein fractions with higher valuable techno-functional properties as well as enriched lipid fractions. The use of different organic acids was evaluated and compared with an inorganic acid (HCl). A beneficial effect of organic acids on the lipid extraction yield (shift from 35 to 45%) was observed, that did not negatively influence the protein extraction efficiency. One organic acid, in particular, lactic acid, increased the lipid purity from 75 to 85%. Protein fractions with a purity of 60% proteins were achieved and were shown to preserve partial solubility. Use of some organic acids at low pH (pH 2) resulted in (1) a higher lipid purity in the lipid-enriched fraction and (2) had similar results in protein fractionation as the inorganic acid. Reproducibility of the fractionation process was proven.

‘Management of waste as a resource’ was described in a European roadmap (COM (2011/571) as a milestone to be reached by 2020. On the one hand, there are diverse under-spent side-streams situated at the farmers and post-harvesting level as well as in the associated retail sector. On the other hand, there is a need for new resources, for instance proteins as alternative for soy proteins. InDIRECT, a three-year EU research project started November 2016, aims to develop biorefinery processes as part of new value chains to convert under-spent side-streams from the agro-sector and processing sector into useful marketable products. Cascading processes (recovery of multiple compounds from the same feedstock) are envisioned to increase the conversion efficiency and maximise the values of the feedstock. Direct bio-refiney of the biomass will be compared with indirect biorefinery. The latter refers to the two-step process that can cope with the heterogeneity of side-streams. In a first step the heterogenic feedstock is converted to homogenous biomass using insects. Insects are able to convert a variety of feedstocks into a more homogenous biomass, being their own biomass. In a second step, the insect biomass is further fractionated into a lipid, protein and chitin fraction, that all three have potential towards marketable end prod-ucts. The direct approach (without the insect conversion step) which is being explored for green leaves, is expected to be more seasonally variable compared to the indirect biorefinery appraoch, but generates a larger variety of compounds. The project foresees to process the derived fractions further into multiple marketable products for the feed, food and chemical sector. The multidisciplinary InDIRECT consortium consists of 9 partners from four countries (Belgium, The Netherlands, France & Italy), comprising 2 research organisations (VITO & University of Parma), 5 industrial partners (Nutrition sciences NV specialized in feed additives, two insect farms Millibeter & Proti-Farm R&D, IMPROVE specialized in plant proteins, and CHEMSTREAM focusing on chemical application), a non-profit organisa-tion that works in close contact with farmers and processing industry (Innovatiesteun-punt) and a project supporting company (Temperio). The project (www.bbi-indirect.eu) is coordinated by VITO. Session 1 Evaluation of the use of insects Bornimer Agrartechnische Berichte I Heft 97 I ISSN 0947-7314 24 This project has received funding from the Bio Based Industries Joint Undertaking under the European Union’s Horizon 2020 research and innovation programme under grant agreement No 720715.

Biodegradation is one of the most favored and sustainable means of removing organic pollutants from contaminated aquifers but the major steering factors are still surprisingly poorly understood. Growing evidence questions some of the established concepts for control of biodegradation. Here, we critically discuss classical concepts such as the thermodynamic redox zonation, or the use of steady state transport scenarios for assessing biodegradation rates. Furthermore, we discuss if the absence of specific degrader populations can explain poor biodegradation. We propose updated perspectives on the controls of biodegradation in contaminant plumes. These include the plume fringe concept, transport limitations, and transient conditions as currently underestimated processes affecting biodegradation.

Recently there has been increased interest in the use of insects for food and feed. This is not only because of the high protein content in insect biomass, as also the lipid frac-tion has value. Chitin, a third major compound in insects (present in the exoskeletons), can be beneficial for feed/food in very low concentrations but affects the digestibility and the utilization of its macro and micro-nutrients in the present concentration. The chitin removal of bee extract for example increased digestibility of the proteins from 71 to 94%. Hence there is a need for separation technologies. The extracted chitin, protein and fat fractions can further be processed for various applications besides food or feed. This poster presents a biorefinery for fractionating insect biomass focusing on preserv-ing the functional properties of proteins. In this way, proteins do not only have nutritional value but can serve as functional ingredient. More specifically, an approach has been elaborated to fractionate larvae of the black soldier fly into chitin, lipids and protein frac-tions envisioning maximal valorization of the biomass for application in feed and chem-istry. These larvae are well-known waste degrading insects, able to grow on different agro-industrial side streams and are on the list of the most promising insects for indus-trial production in the western world. Through the fractionation procedures high-value products like proteins, chitin and lipids can be recovered and thus turning waste into a second-generation resource. Separating fat from animal residue is traditionally performed by rendering, a high tem-perature process resulting in denatured proteins. Our new fractionation procedures will be compared to the rendering process focusing on yield, purity and especially on pre-serving the functional properties of proteins. Furthermore, we will discuss different pre-treatment and separation techniques to remove chitin. Results show that pretreatment as well as the solvent type has a major impact on the fractionation efficiency. This pro-ject has partially received funding from BBI Joint Undertaking under the European Un-ion’s Horizon 2020 research and innovation program under grant agreement No 720715.