Responsiveness of Aromatoleum aromaticum EbN1T to Lignin-Derived Phenylpropanoids.

The betaproteobacterial degradation specialist Aromatoleum aromaticum EbN1T utilizes several plant-derived 3-phenylpropanoids coupled to denitrification. In vivo responsiveness of A. aromaticum EbN1T was studied by exposing nonadapted cells to distinct pulses (spanning 100 µM to 0.1 nM) of 3-phenylpropanoate, cinnamate, 3-(4-hydroxyphenyl)propanoate, or p-coumarate. Time-resolved, targeted transcript analyses via quantitative reverse transcription-PCR of four selected 3-phenylpropanoid genes revealed a response threshold of 30 to 50 nM for p-coumarate and 1 to 10 nM for the other three tested 3-phenylpropanoids. At these concentrations, transmembrane effector equilibration is attained by passive diffusion rather than active uptake via the ABC transporter, presumably serving the studied 3-phenylpropanoids as well as benzoate. Highly substrate-specific enzyme formation (EbA5316 to EbA5321 [EbA5316-21]) for the shared peripheral degradation pathway putatively involves the predicted TetR-type transcriptional repressor PprR. Accordingly, relative transcript abundances of ebA5316-21 are lower in succinate- and benzoate-grown wild-type cells than in an unmarked in-frame ΔpprR mutant. In trans-complementation of pprR into the ΔpprR background restored wild-type-like transcript levels. When adapted to p-coumarate, the three genotypes had relative transcript abundances similar to those of ebA5316-21 despite a significantly longer lag phase of the pprR-complemented mutant (∼100-fold higher pprR transcript level than the wild type). Notably, transcript levels of ebA5316-21 were ∼10- to 100-fold higher in p-coumarate- than succinate- or benzoate-adapted cells across all three genotypes. This indicates the additional involvement of an unknown transcriptional regulator. Furthermore, physiological, transcriptional, and (aromatic) acyl-coenzyme A ester intermediate analyses of the wild type and ΔpprR mutant grown with binary substrate mixtures suggest a mode of catabolite repression of superior order to PprR.IMPORTANCE Lignin is a ubiquitous heterobiopolymer built from a suite of 3-phenylpropanoid subunits. It accounts for more than 30% of the global plant dry material, and lignin-related compounds are increasingly released into the environment from anthropogenic sources, i.e., by wastewater effluents from the paper and pulp industry. Hence, following biological or industrial decomplexation of lignin, vast amounts of structurally diverse 3-phenylpropanoids enter terrestrial and aquatic habitats, where they serve as substrates for microbial degradation. This raises the question of what signaling systems environmental bacteria employ to detect these nutritionally attractive compounds and to adjust their catabolism accordingly. Moreover, determining in vivo response thresholds of an anaerobic degradation specialist such as A. aromaticum EbN1T for these aromatic compounds provides insights into the environmental fate of the latter, i.e., when they could escape biodegradation due to too low ambient concentrations.

Biowaste treatment with black soldier fly larvae: Increasing performance through the formulation of biowastes based on protein and carbohydrates.

A key challenge for black soldier fly larvae (BSFL) treatment is its variable reliability and efficiency when applied to different biowastes. Similar to other biowaste treatment technologies, co-conversion could compensate for variability in the composition of biowastes. Using detailed nutrient analyses, this study assessed whether mixing biowastes to similar protein and non-fibre carbohydrate (NFC) contents increased the performance and reduced the variability of BSFL treatment in comparison to the treatment of individual wastes. The biowastes examined were mill by-products, human faeces, poultry slaughterhouse waste, cow manure, and canteen waste. Biowaste formulations had a protein-to-NFC ratio of 1:1, a protein content of 14-19%, and a NFC content of 13-15% (dry mass). Performance parameters that were assessed included survival and bioconversion rate, waste reduction, and waste conversion and protein conversion efficiency. In comparison to poultry feed (benchmark), vegetable canteen waste showed the best performance and cow manure performed worst. Formulations showed significantly improved performance and lower variability in comparison to the individual wastes. However, variability in performance was higher than expected for the formulations. One reason for this variability could be different fibre and lipid contents, which correlated with the performance results of the formulations. Overall, this research provides baseline knowledge and guidance on how BSFL treatment facilities may systematically operate using biowastes of varying types and compositions.

Black Soldier Fly biowaste treatment - Assessment of global warming potential.

Cities of low and middle-income countries face severe challenges in managing the increasing amount of waste produced, especially the organic fraction. Black Soldier Fly (BSF) biowaste treatment is an attractive treatment option as it offers a solution for waste management while also providing a protein source to help alleviate the rising global demand for animal feed. However, to-date very little information is available on how this technology performs with regard to direct greenhouse gas (GHG) emissions and global warming potential (GWP). This paper presents a study that uses a life cycle assessment (LCA) approach to assess the GWP of a BSF waste treatment facility in the case of Indonesia and compares it with respective values for an open windrow composting facility. Direct CH4 and N2O samples were extracted from BSF treatment units and analyzed by gas chromatography. Results show that direct CO2eq emissions are 47 times lower the emissions from composting. Regarding the overall GWP, the LCA shows that composting has double the GWP of BSF treatment facility based on the functional unit of 1 ton of biowaste (wet weight). The main GWP contribution from a BSF facility are from: (1) residue post-composting (69%) and (2) electricity needs and source (up to 55%). Fishmeal production substitution by BSF larvae meal can reduce significantly the GWP (up to 30%). Based on this study, we conclude that BSF biowaste treatment offers an environmentally relevant alternative with very low direct GHG emissions and potentially high GWP reduction. Further research should improve residue post-treatment.

Decomposition of biowaste macronutrients, microbes, and chemicals in black soldier fly larval treatment: A review.

Processing of biowaste with larvae of the black soldier fly, Hermetia illucens L. (Diptera: Stratiomyidae), is an emerging waste treatment technology. Larvae grown on biowaste can be a relevant raw material for animal feed production and can therefore provide revenues for financially viable waste management systems. In addition, when produced on biowaste, insect-based feeds can be more sustainable than conventional feeds. Among others, the scalability of the technology will depend on the availability of large amounts of biowaste with a high process performance (e.g. bioconversion of organic matter to proteins and lipids) and microbial and chemical product safety. Currently, in contrast to other waste treatment technologies, such as composting or anaerobic digestion, the process performance is variable and the processes driving the decomposition of biowaste macronutrients, inactivation of microbes and fate of chemicals is poorly understood. This review presents the first summary of the most important processes involved in black soldier fly larvae (BSFL) treatment, based on the available knowledge concerning five well-studied fly species. This is a starting point to increase understanding regarding the processes of this technology, with the potential to increase its efficiency and uptake, and support the development of appropriate regulations. Based on this review, formulating different types of biowaste, e.g. to produce a diet with a similar protein content, a balanced amino acid profile and/or pre- and co-treatment of biowaste with beneficial microbes, has the potential to increase process performance. Following harvest, larvae require heat or other treatments for microbial inactivation and safety.

Faecal sludge management with the larvae of the black soldier fly (Hermetia illucens)--from a hygiene aspect.

Inadequate and lacking sanitation and wastewater treatment systems can lead to the spreading of diarrhoeal diseases. One contributing factor in the lack of such treatment systems is the lack of economic incentives for stakeholders throughout the service chain. However, the organic fraction of the waste is high in valuable plant nutrients and could be reused in agriculture and as animal feed. For example, grown larvae of the black soldier fly, Hermetia illucens L. (Diptera: Stratiomyidae), make an excellent protein source in animal feed, while the feeding activity of the larvae substantially reduces the dry mass of the treated material. This study examined the effect of black soldier fly larvae on the concentration of pathogenic microorganisms in human faeces and found a 6 log10 reduction in Salmonella spp. in human faeces in eight days, compared with a <2 log10 reduction in the control. No increased reduction was observed for Enterococcus spp., bacteriophage ΦX174 or Ascaris suum ova.

Conversion of organic material by black soldier fly larvae: establishing optimal feeding rates.

Larvae of the black soldier fly, Hermetia illucens (Diptera: Stratiomyidae), are voracious feeders of organic material and may thus be used in simple engineered systems to reduce organic waste in low- and middle-income countries. Controlled feeding experiments with standard fodder were conducted to assess the optimum amount of organic waste to be added to a CORS system (Conversion of Organic Refuse by Saprophages). A daily feeding rate of 100 mg chicken feed (60% moisture content) per larva resulted in an optimum trade-off between material reduction efficiency (41.8%, SE 0.61) and biomass production (prepupal dry weight: 48.0 mg, SE 2.0). Applied to market waste and human faeces, this corresponds to a potential daily feeding capacity of 3-5 kg/m(2) and 6.5 kg/m(2), respectively. In addition, H. illucens prepupae quality was assessed to determine their suitability to substitute fishmeal in animal feed production. The chitin-corrected crude protein content ranged from 28.2 to 42.5%, depending on the amount of food provided to the larvae. Based on our study, a waste processing unit could yield a daily prepupal biomass of 145 g (dry mass) per m(2). We conclude that larvae of the black soldier fly are potentially capable of converting large amounts of organic waste into protein-rich biomass to substitute fishmeal, thereby contributing to sustainable aquaculture.