A comparison in product‐value potential in four treatment strategies for food waste and faeces – assessing composting,fly larvae composting and anaerobic digestion

Municipalities are expected to provide solid waste management, which is funded by tax revenue or/and waste treatment fees. In many low‐ and middle‐income countries, municipalities struggle to provide an adequate level of service, and in these places, the informal sector plays a major role in the collection and treatment of solid waste. In contrast to the plastic and metal fraction, the organic fraction is not managed by the informal sector, primarily because it has low or no financial value and treatment would cost more than the possible revenue. If the organic fraction could be converted to valuable products, the treatment could bear its own cost and this could act as an incentive to collect and treat this fraction. In this study, the potential product value generated through four treatment strategies treating food waste and faeces was compared in a Swedish context: (i) thermophilic composting; (ii) black soldier fly treatment (BSF treatment); (iii) anaerobic digestion (AD); and (iv) BSF treatment followed by AD (BSF + AD). In order to assess the AD strategies, the biomethane potentials of the substrates were assessed. Food waste had the highest biomethane potential, while BSF‐treated faeces had the lowest (417 and 188 NmL g VS^?1, respectively). Thermophilic composting yielded the lowest value product (organic fertilizer; 26 € t^?1 treated food waste) and BSF treatment + AD the highest total value of products (animal feed, vehicle gas and organic fertilizer; 215 € t^?1 treated food waste). The treatment costs were not taken into account here; the total value gives an indication of the cost margin for the different strategies studied. In places with an existing AD plant, BSF treatment + AD strategy is the most economically viable. In places where no such plant exists, BSF treatment is likely to be the most economically favourable treatment.     

Improvement in bioconversion efficiency and reduction of ammonia emission by introduction of fruit fermentation broth in a black soldier fly larvae and kitchen waste conversion system

The black soldier fly (BSF), Hermetia illucens (Diptera: Stratiomyidae), is an insect commonly used for the bioconversion of various organic wastes. Not only can the BSF convert organic waste into macromolecular organic substances, such as insect proteins, but it can also lessen the pollution associated with these waste products by reducing ammonia emissions, for example. In this study, we measured the effects of adding fruit fermentation broth (Fer) and commercial lactic acid bacteria fermentation broth (Em) to kitchen waste (KW), as deodorizing auxiliary substances, on the growth performance of black soldier fly larvae (BSFL), the intestinal flora structure of BSFL, the ammonia emission from the KW substrate, and the microbial community structure of the KW substrate. We found that the addition of Fer or Em increased the body weight of BSFL after 6 d of culture, increasing the growth rate by 9.96% and 7.96%, respectively. The addition of Fer not only reduced the pH of the KW substrate but also increased the relative abundance of probiotics, such as Lactobacillus, Lysinibacillus, and Vagococcus, which inhibited the growth of ammonifiers such as Bacillus, Oligella, Paenalcaligenes, Paenibacillus, Pseudogracilibacillus, and Pseudomonas, resulting in the reduction of ammonia emission in the KW substrate. Moreover, the addition of Fer or Em significantly increased the relative abundances of Bacteroides, Campylobacter, Dysgonomonas, Enterococcus, and Ignatzschineria in the gut of BSFL and increased the species diversity and richness in the KW substrate. Our findings provide a novel way to improve the conversion rate of organic waste and reduce the environmental pollution caused by BSF.     

Establishment of highly efficient transgenic system for black soldier fly (Hermetia illucens)

The black soldier fly (BSF), Hermetia illucens, is a promising insect for mitigating solid waste problems as its larvae are able to bioconvert organic waste into valuable biomass. We recently reported a high-quality genome assembly of the BSF; analysis of this genome sequence will further the understanding of insect biology and identify genes that can be manipulated to improve efficiency of bioconversion. To enable genetic manipulation of the BSF, we have established the first transgenic methods for this economically important insect. We cloned and identified the ubiquitous actin5C promoter (Hiactin5C-p3k) and 3 endogenous U6 promoters (HiU6:1, HiU6:2, and HiU6:3). The Hiactin5C promoter was used to drive expression of a hyperactive variant of the piggyBac transposase, which exhibited up to 6-fold improvement in transformation rate when compared to the wild-type transposase. Furthermore, we evaluated the 3 HiU6 promoters using this transgenic system. HiU6:1 and HiU6:2 promoters provided the highest knockdown efficiency with RNAi and are thus promising candidates for future Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) development. Overall, our findings provide valuable genetic engineering toolkits for basic research and genetic manipulation of the BSF.     

The Intestinal Bacterial Community in the Food Waste-Reducing Larvae of <Emphasis Type="Italic">Hermetia illucens</Emphasis>

As it is known that food waste can be reduced by the larvae of Hermetia illucens (Black soldier fly, BSF), the scientific and commercial value of BSF larvae has increased recently. We hypothesised that the ability of catabolic degradation by BSF larvae might be due to intestinal microorganisms. Herein, we analysed the bacterial communities in the gut of BSF larvae by pyrosequencing of extracting intestinal metagenomic DNA from larvae that had been fed three different diets. The 16S rRNA sequencing results produced 9737, 9723 and 5985 PCR products from larval samples fed food waste, cooked rice and calf forage, respectively. A BLAST search using the EzTaxon program showed that the bacterial community in the gut of larvae fed three different diets was mainly composed of the four phyla with dissimilar proportions. Although the composition of the bacterial communities depended on the different nutrient sources, the identified bacterial strains in the gut of BSF larvae represented unique bacterial species that were unlike the intestinal microflora of other insects. Thus, our study analysed the structure of the bacterial communities in the gut of BSF larvae after three different feedings and assessed the application of particular bacteria for the efficient degradation of organic compounds.     

Hermetia illucens L. larvae–associated intestinal microbes reduce the transmission risk of zoonotic pathogens in pig manure

Black soldier fly (BSF) larvae are considered a promising biological reactor to convert organic waste and reduce the impact of zoonotic pathogens on the environment. We analysed the effects of BSF larvae on Staphylococcus aureus and Salmonella spp. populations in pig manure (PM), which showed that BSF larvae can significantly reduce the counts of the associated S. aureus and Salmonella spp. Then, using a sterile BSF larval system, we validated the function of BSF larval intestinal microbiota in vivo to suppress pathogens, and lastly, we isolated eight bacterial strains from the BSF larval gut that inhibit S. aureus. Results indicated that functional microbes are essential for BSF larvae to antagonise S. aureus. Moreover, the analysis results of the relationship between the intestinal microbiota and S. aureus and Salmonella spp. showed that Myroides, Tissierella, Oblitimonas, Paenalcalignes, Terrisporobacter, Clostridium, Fastidiosipila, Pseudomonas, Ignatzschineria, Savagea, Moheibacter and Sphingobacterium were negatively correlated with S. aureus and Salmonella. Overall, these results suggested that the potential ability of BSF larvae to inhibit S. aureus and Salmonella spp. present in PM is accomplished primarily by gut-associated microorganisms.     

Nutritional composition of black soldier fly larvae feeding on agro-industrial by-products

Black soldier fly (BSF) larvae, Hermetia illucens L. (Diptera: Stratiomyidae), bio-convert organic side streams into high-quality biomass, the composition of which largely depends on the side stream used. In the present study, BSF larvae were reared on feed substrates composed of dried brewers’ spent grains, each supplemented with either water, waste brewer’s yeast, or a mixture of waste brewer’s yeast and cane molasses to obtain 12 different substrates: barley/water, barley/yeast, barley/yeast/molasses, malted barley/water, malted barley/yeast, malted barley/yeast/molasses, malted corn/water, malted corn/yeast, malted corn/yeast/molasses, sorghum-barley/water, sorghum-barley/yeast, and sorghum-barley/yeast/molasses. The crude protein, fat, ash, and mineral contents of the BSF larvae fed each feed substrate were quantified by chemical analyses. The effect of substrate, supplementation, and their interaction on crude protein, fat, and ash contents of BSF larval body composition was significant. Calcium, phosphorus, and potassium were the most abundant macrominerals in the larvae and their concentrations differed significantly among substrates. These findings provide important information to support the use of BSF larval meal as potential new source of nutrient-rich and sustainable animal feed ingredients to substitute expensive and scarce protein sources such as fishmeal and soya bean meal.     

Review of insect pathogen risks for the black soldier fly (Hermetia illucens) and guidelines for reliable production

Black soldier fly [BSF; Hermetia illucens L. (Diptera: Stratiomyidae)] larvae are very effective in transforming low-grade food waste into valuable high-end proteins and fat, in intensive production facilities. The production output of this species is growing quickly, but upscaling brings risks to the health status of the reared insects. Until now, not a single major case of disease outbreak caused by a pathogen in a BSF production unit has been reported. This contrasts with data on other species of mass-produced insects, which have experienced various disease outbreaks, indicating that BSFs are comparatively resistant to insect diseases. Further, there are no records of natural infections caused by entomopathogens in BSF. In this review, the known entomopathogens of Diptera, especially BSF, and their potential risks for causing disease in these insects are summarized.     

Dynamics of the intestinal bacterial community in black soldier fly larval guts and its influence on insect growth and development

Black soldier fly (BSF), Hermetia illucens (Diptera: Stratiomyidae), is a prominent insect for the bioconversion of various organic wastes. As a saprotrophic insect, the BSF inhabits microbe-rich environments. However, the influences of the intestinal microorganisms on BSF growth and development are not very clear. In this study, the dynamics of the intestinal bacterial community of BSF larvae (BSFL) were analyzed using pyrosequencing. Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria were the most prevalent bacterial phyla in the intestines of all larval instars. The dynamic changes in bacterial community compositions among different larval instars were striking at the genus level. Klebsiella, Clostridium, Providencia, and Dysgonomonas were the relatively most abundant bacteria in the 1st- to 4th-instar BSFL, respectively. Dysgonomonas and Providencia also dominated the 5th- and 6th-instar larvae, at ratios of 31.1% and 47.2%, respectively. In total, 148 bacterial strains affiliated with 20 genera were isolated on different media under aerobic and anaerobic conditions. Among them, 6 bacteria, BSF1–BSF6, were selected for further study. The inoculation of the 6 isolates independently into germ-free BSFL feeding on an artificial diet showed that all the bacteria, except BSF4, significantly promoted BSF growth and development compared with the germ-free control. Citrobacter, Dysgonomonas, Klebsiella, Ochrobactrum, and Providencia promoted BSF development significantly by increasing the weight gains of larvae and pupae, as well as increasing the prepupae and eclosion rates. In addition, Citrobacter, Klebsiella and Providencia shortened the BSF life cycle significantly. The results illustrate the promotive effects of intestinal bacteria on BSF growth and development.     

Potential of Allolobophora parva (Oligochaeta) in vermicomposting

The aim of this study was to assess the potential of Allolobophora parva Eisen as a candidate for vermicomposting practices. Five organic waste mixtures: cow dung (CD), biogas plant slurry (BGS), cow dung + vegetable waste (CD + VW), BGS + VW and VW + Soil were vermicomposted using A. parva. Vermicomposting showed a decrease in pH, organic C and C:N ratio, but increase total N, available P and exchangeable K at the end. C:N ratio of end material (vermicompost) was within the agronomic acceptable limit (<20). The high level of NPK in worm-processed material indicates the candidature of this species for waste management operations. The earthworm also showed an excellent growth in different wastes. Results thus indicate that A. parva appeared a potential tool for conversion of organic wastes into value added products for sustainable land restoration practices.     

Distribution and diversity of diatom assemblages in surficial sediments of shallow lakes in Wapusk National Park (Manitoba,Canada) region of the Hudson Bay Lowlands

The hydrology of shallow lakes (and ponds) located in the western Hudson Bay Lowlands (HBL) is sensitive to climate warming and associated permafrost thaw. However, their biological characteristics are poorly known, which hampers effective aquatic ecosystem monitoring. Located in northern Manitoba along the southwestern coast of Hudson Bay, Wapusk National Park (WNP) encompasses numerous shallow lakes representative of the subarctic zone. We analyzed the distribution and diversity of diatom (microscopic algae; class Bacillariophyceae) assemblages in surficial sediments of 33 lakes located in three different ecozones spanning a vegetation gradient, from NE to SW: the Coastal Fen (CF), the Interior Peat Plateau (IPP), and the Boreal Spruce Forest (BSF). We found significant differences (P < 0.05) in diatom community composition between CF and IPP lakes, and CF and BSF lakes, but not between IPP and BSF lakes. These results are consistent with water chemistry measurements, which indicated distinct limnological conditions for CF lakes. Diatom communities in CF lakes were generally dominated by alkaliphilous taxa typical of waters with medium to high conductivity, such as Nitzschia denticula. In contrast, several IPP and BSF lakes were dominated by acidophilous and circumneutral diatom taxa with preference for low conductivity (e.g., Tabellaria flocculosa, Eunotia mucophila, E. necompacta var. vixcompacta). This exploratory survey provides a first detailed inventory of the diatom assemblages in the WNP region needed for monitoring programs to detect changes in shallow lake ecosystems and ecozonal shifts in response to climate variations.     

Effects of periodic administration ofNocardia rubra cell-wall skeleton on immunoglobulin production and B-cell-stimulatory factor activity in vitro in workers at a poison gas factory

Summary The former workers at the Okunojima poison gas factory (poison gas workers) are a high-risk group for malignant neoplasms and show abnormalities in cellular immunity. At the same time, poison gas workers often have chronic respiratory diseases, such as chronic bronchitis, and are highly susceptible to respiratory infections. To explore the possibility of immunological cancer prevention, we have periodically administered 200 µgNocardia rubra cell-wall skeleton (N-CWS) to poison gas workers once every 3 months since December 1978. During this period, we noted a significantly lower incidence of influenza among poison gas workers receiving N-CWS than in those not receiving the drug during the influenza epidemic. This finding suggested that the administration of N-CWS enhanced the resistance of these workers to infections. Therefore, periodical administration of N-CWS to poison gas workers was considered to enhance the reduced T-cell function of normalizing antibody production by stimulating the production of B-cell-stimulatory factor (BSF). In the present study, to clarify the mechanism of immunosuppression in the poison gas workers and to examine the effects of continual administration of N-CWS on this condition, we compared the immunoglobulin production and the proliferative and differentiative activities of B-cell-stimulatory factor (BSF) of peripheral blood mononuclear cells (PBMC), in poison gas workers treated or not treated with N-CWS. Comparisons were also made with age-matched healthy controls. In the untreated poison gas workers, immunoglobulin and BSF production of PBMC were reduced as compared with the control group. On the other hand, in the poison gas workers receiving N-CWS, immunoglobulin and BSF production of PBMC were restored nearly to the control level. These results show that in vitro antibody production in the poison gas workers was reduced and that a reduction in BSF production of T cells was one of its causes.     

Response of an estuarine ecosystem to reduced organic waste discharge

Since the mid-19^th century large amounts of organic wastewere sluiced out into the Ems Estuary on the border between Germany and TheNetherlands. This waste originated from the regional potato flour and cardboardindustries making the inland waterways completely anoxic. In the estuaryseriousoxygen depletion occurred, especially during autumn. Most of the organic wasteentered the estuary in the Dollard, a brackish embayment of the Ems Estuary. Anintensive sanitation scheme was started in the 1970s, leading to a stepwisereduction of the organic waste load on the estuary. In this paper, a review isgiven of the response of the benthos living at intertidal mudflats,viz. microphytobenthos (diatoms), meiofauna (nematodes)andmacrozoobenthos. The benthos response is described mainly on the basis of dataobtained under conditions of high (ca. 1980), intermediate (1987) and largelyreduced (1993) organic waste loading.Reduction of organic loading caused significant changes in abundance, speciescomposition and standing stock of diatoms and nematodes. Macrobenthicpopulations recovered from being severely reduced regularly during the autumnalwaste discharges towards a more stable situation. Altogether, the intertidalmudflat benthos changed from organic waste loading stress towards a normal,estuarine environmental stress. This was especially the case at high and muddyflats in the southeast of the Dollard. At mixed sand/mud flats in the centre ofthe Dollard, a response of macrozoobenthos may have been obscured by theinvasion of the polychaete Marenzelleria cf.wireni.     

Preliminary evaluation of fungicidal and termiticidal activities of filtrates from biomass slurry fuel production

Biomass slurry fuel (BSF) production has recently been developed as a natural energy for the conversion of solid biomass into fuel. In addition to using fuel, filtrates from BSF production may also serve a chemical source with several organic compounds. There is an increasing interest in the research and application of biomass-based filtrates. In this study, fungicidal and termiticidal properties of filtrates from BSF production using sugi (Cryptomeria japonica) and acacia (Acacia mangium) wood were evaluated in laboratory decay and termite resistance tests. Wood blocks treated with the filtrates showed increased resistance against brown-rot fungus, Fomitopsis palustris. However the filtrates from sugi wood processed at 270 degrees C which contained less phenolic compounds than the other filtrates were effective against white-rot fungus, Trametes versicolor. Phenolic compounds of filtrates seemed to play a role in the decay resistance tests however the filtrates did not increase the durability of the wood blocks against subterranean termites Coptotermes formosanus. Despite high acetic and lactic acid content of the filtrates, vanillin content of the filtrates may have served as an additional food source and promoted termite attack. It can be concluded that filtrates with phenolic compounds from lignin degradation during BSF production can be considered for targeted inhibition of brown-rot.     

黑水虻幼虫粉对凡纳滨对虾生长、免疫和脂质代谢的影响

试验旨在研究黑水虻Hermetia illucens幼虫粉替代鱼粉对凡纳滨对虾Litopenaeus vannamei生长性能、非特异性免疫和脂质代谢的影响。以黑水虻幼虫粉分别替代对照组饲料(FM)中10%(BSF10)、20%(BSF20)和30%(BSF30)的鱼粉蛋白质,共配制为四组等氮等脂的实验饲料,饲喂初始体质量为(0.88±0.01) g的凡纳滨对虾7周。结果显示, BSF10组和BSF20组对虾的生长性能与对照组相比无显著变化(P>0.05),但BSF30组对虾的生长性能显著降低(P<0.05)。与对照组相比, BSF20组和BSF30组对虾的全虾粗脂肪含量显著降低,BSF30组对虾血淋巴甘油三酯和总胆固醇水平显著降低(P<0.05),但肝胰腺粗脂肪水平无显著差异(P>0.05)。BSF10、BSF20和BSF30组对虾血淋巴谷丙转氨酶和谷草转氨酶活性显著低于对照组(P<0.05)。BSF10组对虾超氧化物歧化酶活性显著高于对照组, BSF20组对虾总抗氧化能力显著高于其余各组(P<0.05)。BSF30组对虾肝胰腺酸性磷酸酶和碱性磷酸...     

Effects of pod infection by Mycosphaerella pinodes on yield components of pea (Pisum sativum)

Yield reduction of pea (Pisum sativum) due to various types of infections by Mycosphaerella pinodes on pods was assessed. A range of disease severities was created on pods of pea plants grown in the glasshouse, by painting the pods with different concentrations of spore suspensions, at three different pod development stages: lag phase, the beginning of seed filling (BSF) and mid-filling of the seeds. Seed number at harvest was reduced only if the pods were infected before BSF, as shown previously for whole plant infections. Pod infections led to individual seed weight (ISW) losses from zero (for late infections, at mid-filling) to 20% (for earlier infections and severe disease). Infection during the lag phase affected ISW by reducing seed growth rate, whereas infection at BSF tended to reduce the duration of seed filling. There was a linear relationship between the area under the disease progress curve and the percentage decrease in ISW. This model should be complemented by the effect of leaves and stem infections, in order to predict ISW losses in diseased crop conditions, in which epidemics occur on all aerial parts of the pea plant.     

Enzyme Activities and Microflora of Earthworm Gut and Vermireactors as Indicators of the Stabilization of Waste Degradation Process

The worm and microorganisms act to enhance the biodegradation of organic matter. In earlier studies effect of enzymes has been studied in waste degradation process; however, no attempt has been made to determine the end point of organic waste degradation process through indication of hydrolases and dehydrogenase enzyme activity as well as microflora of earthworm and vermireactors. The present work is a pioneer endeavor in this direction. The study indicates that with time, hydrolase (β-glucosidase, N-x-benzoyl-l-argininamide [BAA]-hydrolyzing protease, urease, phosphatase, and cellulase) and dehydrogenase activities decrease as available organic compounds decreased. A high correlation among all enzyme activity and microflora was observed that led to the conclusion that activities of both enzymes (hydrolytic and dehydrogenase) and microflora could be the feasible indicators of organic matter degradation process.     

Ultrastructural Changes of the Mitochondrion During the Life Cycle of Trypanosoma brucei

The mitochondrion is crucial for ATP generation by oxidative phosphorylation, among other processes. Cristae are invaginations of the mitochondrial inner membrane that house nearly all the macromolecular complexes that perform oxidative phosphorylation. The unicellular parasite Trypanosoma brucei undergoes during its life cycle extensive remodeling of its single mitochondrion, which reflects major changes in its energy metabolism. While the bloodstream form (BSF) generates ATP exclusively by substrate-level phosphorylation and has a morphologically highly reduced mitochondrion, the insect-dwelling procyclic form (PCF) performs oxidative phosphorylation and has an expanded and reticulated organelle. Here, we have performed high-resolution 3D reconstruction of BSF and PCF mitochondria, with a particular focus on their cristae. By measuring the volumes and surface areas of these structures in complete or nearly complete cells, we have found that mitochondrial cristae are more prominent in BSF than previously thought and their biogenesis seems to be maintained during the cell cycle. Furthermore, PCF cristae exhibit a surprising range of volumes in situ, implying that each crista is acting as an independent bioenergetic unit. Cristae appear to be particularly enriched in the region of the organelle between the nucleus and kinetoplast, the mitochondrial genome, suggesting this part has distinctive properties.     

Microbial diversity in support of anaerobic biomass valorization

Microbial diversity provides an immense reservoir of functions and supports key steps in maintaining ecosystem balance through matter decomposition processes and nutrient recycling. The use of microorganisms for biomolecule production is now common, but often involves single-strain cultures. In this review, we highlight the significance of using ecosystem-derived microbial diversity for biotechnological researches. In the context of organic matter mineralization, diversity of microorganisms is essential and enhances the degradation processes. We focus on anaerobic production of biomolecules of interest from discarded biomass, which is an important issue in the context of organic waste valorization and processing. Organic waste represents an important and renewable raw material but remains underused. It is commonly accepted that anaerobic mineralization of organic waste allows the production of diverse interesting molecules within several fields of application. We provide evidence that complex and diversified microbial communities isolated from ecosystems, i.e. microbial consortia, offer considerable advantages in degrading complex organic waste, to yield biomolecules of interest. We defend our opinion that this approach is more efficient and offers enhanced potential compared to the approaches that use single strain cultures.     

Estuarine nematodes as indicators of organic pollution; An example from the Ems estuary (The Netherlands)

A comparison is made of the structure and abundance of the nematode community of an intertidal, brackish-water mudflat in the Ems estuary under conditions of high (1980) and reduced (1987) organic waste input. Three stations, at different distances from the waste water inlet, were sampled monthly; nematodes were identified and counted in the upper 1 cm of the sediment. In 1987, numerical density of nematodes was lower, and species diversity was higher than in 1980. Diatom eating nematode species dominated in 1980, to decrease by 1987.Eudiplogaster pararmatus, an indicator of organic enrichment, also decreased strongly. The response of the nematode community to reduced organic waste input could be adequately described with the BONGERS' Maturity Index.