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With a growing human population, food insecurity is becoming a worldwide problem. As the search for sustainable sources of protein continues, black soldier fly larvae come to the forefront as a partial solution. Full of proteins and fats, the larvae will consume most organic matter and rapidly develop into a usable form. Supplementing black soldier fly larvae with oleaginous microbes Arthrobacter AK19 and Rhodococcus rhodochrous increases their body size by 25%, potentially accelerates their development by one day, and increases their conversion efficiency. Supplementing with Bifidobacterium breve decreased body size, slowed development, and decreased conversion efficiency, underscoring the importance of selecting supplemental microbes and testing first on a small-scale. Promising results on the small-scale led to an industrial study, where similar results were also seen, resulting in greater optimization of this system.

Microaerobic fermentation (MF) is a process of controlled degradation of organic waste material that occurs in enclosed fermentors under micro-aerobic conditions at near-room temperature. MF processing of vegetal materials progresses to endpoints in about 2-5 weeks. During MF processing, an acidic leachate rich in organic acids and alcohols is produced. The research presented in this thesis focuses on the efficiency of MF pre-processing of feedstock containing fibrous lignocellulosic (FLC) materials; efficiency of microbial and insect larvae-based treatments of MF leachate; tolerance of the Black Soldier fly larvae (BSFL) to various biological inhibitors common in leachate; and effectiveness of using MF and BSFL solid and liquid processing products as agricultural fertilizers. Results indicate that MF is unsuitable for pre-processing of FLC materials. Enhanced MF leachate treatment may increase efficiency of FLC processing though. Leachate can be efficiently treated using BSFL which decrease overall leachate toxicity. BSFL are able to tolerate increased levels of many of the biological inhibitors within the leachate including ethanol, acetate, pH extremes and temperature. MF solid residues increased corn plant growth when amended into soil, but residues resulting from BSFL processing of solid organics stunted corn plant growth. Short-term phytotoxicity of MF leachate was eliminated by diluting it 10 - 10,000 times or through BSFL processing. It can be concluded that MF processing of organics is beneficial for producing solid soil amendments from non-FLC materials and that dilution or BSFL treatment of MF leachate leads to a beneficial liquid fertilizer.

<p>  </p> <p>Urban farmers face many unique challenges associated with the urban environment in which they produce. One of the most expensive and limited resources is access to healthy soils. There is often low organic matter and industrial contaminants present in urban soils, resulting in the need for remediation, such as capping and importing topsoil and compost. Recently, black soldier fly larvae (<em>Hermetia illucens; BSF)</em> have been recognized as an efficient organism used to break-down organic matter and produce a soil amendment comparable to traditional fertilizers. These fly larvae can feed on a wide range of organic waste (plant material, biosolids, food waste, etc.), can break down contaminants such as pharmaceuticals or pesticides, and impact the bioavailability of heavy metals. The resulting material is a digestate that can be applied as a soil amendment, much like the vermicomposting processes of worms. Fly pupae can be harvested and used as a nutrient dense feedstock for livestock or reared to adults to continue the cycle of composting.  Knowledge gaps remain regarding the impact of feedstock on the nutritional quality of the digestate for crop production and the application and implementation of BSF composting on-farm. We found that larval weight is unaffected by diet streams, however, larval length is improved on food waste streams. Additionally, crop growth varies when grown with BSFL digestate.</p>

Philosophiae Doctor - PhD / The Black soldier fly (BSF) is an environmentally friendly and sustainable insect utilised in the decomposition of organic waste. This is due to its voracious consumption capability, disruptive functions and economic importance. The sustained global increase in commercial BSF farming has resulted in an expanded waste generation from its carcases to which beneficial uses ought to be developed. This study focused on the beneficial use of the generated waste by extracting chitosan from waste pupae and commercially reared BSF adult carcases. The study also considered the conversion of the extracted chitosan to nanofibres and nanoparticles for application in adsorption of inorganic Pb2+ or Cd2+ and antimicrobial studies, respectively. To achieve the aim of this study, the optimal extraction conditions of chitin and chitosan from both pupal exuviae and adult BSF waste materials were attained after a series of experiments. The extraction process involved three stages which were demineralisation, deproteination and deacetylation. The extracted adult and pupal chitin and chitosan were characterised using Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), X-ray diffraction studies (XRD), high resolution scanning electron microscopy (HRSEM) and solid-state carbon nuclear magnetic resonance spectroscopy (13C NMR). Additionally, the adult (ACH20_9) and pupal (PCH21_9) chitosan samples, due to their solubility, were further characterised to determine their molecular weight, fat and water binding capacities, solubility and ash contents. / 2021-09-30

The global demand for animal proteins is projected to rise by 14% by 2030, amplifying the environmental toll of conventional animal-based protein production. Cultivated meat technology can alleviate the growing demand for protein and address the environmental and ethical concerns associated with conventional livestock farming. However, it faces a critical challenge: the high cost of cell culture media, primarily due to the use of Fetal Bovine Serum (FBS). Substituting serum with protein hydrolysates reduces the production expense of cultivated meat products and promotes establishing a sustainable food system. This study explores black soldier fly larvae (Hermetia illucens) as an emerging ethical and cost-effective alternative protein source to replace serum in media, particularly for cultivated meat production. The development of BSFL protein isolate involved defatting the larva, followed by protein extraction. The protein isolate was then hydrolyzed using an enzyme to produce BSFL hydrolysates. The goal was to supplement the protein isolate and hydrolysates with a serum-free media (B8) and determine their efficacy in replacing the 20% serum requirement for the cell culture of Bovine Satellite Cells. The BSFL protein isolate developed had a crude protein content of 80.42% and an amino acid composition conducive to cell proliferation. Experimental concentrations, ranging from 0.006 mg/ml for hydrolysate to 0.06 mg/ml for protein isolate, exhibited enhanced cell growth. Data from dsDNA quantification revealed no significant difference in growth between cells fed serum-containing growth media (BSC-GM) and BSFL protein hydrolysate (BSFLH_1h) over a short-term study. Results from the multi-passage growth study revealed that BSFLH_1h significantly improved cell growth compared to B8 over 4 passages. However, its doubling time was slower than BSC-GM. Additionally, it was observed that the protein isolate and hydrolysate were cytotoxic at higher concentrations. In the future, identifying and removing the cytotoxic compounds can further optimize the media composition. Immunostaining using Pax7 and DAPI identified supplemented media-maintained satellite cell identity of Bovine satellite cells, offering crucial insights into cellular proliferation. Furthermore, since each cell type requires varying serum and nutrients, testing these isolates and hydrolysates on different cell lines can provide better insight into creating a universal serum-free media. / Master of Science in Life Sciences / The global demand for animal proteins is projected to rise by 14% by 2030, amplifying the environmental toll of conventional animal-based protein production. Meat, dairy, aquaculture, and eggs significantly contribute to food-related emissions and occupy a vast portion of global farmland. Cultivated meat production can alleviate the growing demand for protein and address the environmental and ethical concerns associated with conventional livestock farming. Currently, the production of cultivated meat faces a significant hurdle: the high cost of culture media, primarily attributed to the use of Fetal Bovine Serum (FBS). Substituting serum with protein isolates or hydrolysates reduce the production expense of cultivated meat products and promotes a sustainable food system. Protein isolate and hydrolysates derived from black soldier fly larvae (Hermetia illucens) are rich in protein and essential amino acids and can be used as a cost-effective alternative to serum in cell culture media. The protein isolate and hydrolysates derived from BSFL were tested as supplements to a serum-free media (B8) to evaluate their effectiveness in supporting the growth of Bovine Satellite Cells. The protein hydrolysate demonstrated enhanced cell growth at experimental concentrations. However, it could not completely replace serum requirements without slowing cell growth. Despite challenges such as cytotoxicity at higher concentrations, our study suggests that further refinements and application on various cell types can assist in creating a sustainable and affordable serum-free media for cultivated meat production.

Submitted by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-05-11T15:25:55Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação Final_Nelson Poli.pdf: 2785029 bytes, checksum: 10efd62a077b9a359e6ed7401d9664ce (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2018-05-11T15:26:05Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação Final_Nelson Poli.pdf: 2785029 bytes, checksum: 10efd62a077b9a359e6ed7401d9664ce (MD5) / Made available in DSpace on 2018-05-11T15:26:05Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação Final_Nelson Poli.pdf: 2785029 bytes, checksum: 10efd62a077b9a359e6ed7401d9664ce (MD5) Previous issue date: 2018-03-28 / Among the problems explosive world population growth brings to the environment, this work proposes one possible solution to mitigate two problems: disposal of organic solid waste, and the pressure on the current supply of food protein. This solution is based on mass production of Hermetia Illucens larvae (L., 1758) (Diptera: Stratiomyidae), also known as black soldier fly, to degrade organic solid waste, and also serve as an alternative source of animal protein. The optimization of this solution lies on understanding the biology of the insect to find out the relation of the quality and optimal amounts of organic matter to ensure its degradation, and enhance the larvae growth. Finally, knowing the quality of the larvae protein is fundamental to finding its consumer market. To understand these questions, quantitative experiments and analyzes were made. The organic solid residues used as food for the larvae showed in their centesimal composition 66% of humidity, 63,8% of carbohydrates, 13,8% of crude protein and 18,1% of fat. For the mass production of Hermetia Illucens larvae, the study sought to understand its life cycle in the Amazonian biome in comparison to the existing literature. In relation to the larvae biology, it was possible to observe an oviposition rate around 1002 eggs/female. The mating time takes about 15 minutes. It was observed a cycle of about 36 days from oviposition to adulthood. The eggs hatched on average at 74,75 hours. The development time of the larva from egg to the pre-pupae phase took around 21 days and pupation time 17,5 days. 74,48% of the egg reach the adult stage with a sex ratio of 3 males by 1 female. Regarding the supply of the optimum quantity of food to be supplied, it was concluded that for each 100 mg of Hermetia Illucens eggs, the supply of 2000g of restaurant waste in 2 fractions of 1000g with a 5-day feeding interval reached the best potential of dry. It was observed a organic solid waste weight reduction of 83,75% with a conversion rate of solid residue in proteic biomass of 23,2%. The pre-pupal as raw material for the production of animal protein showed 40% of crude protein and 32,6% of fat, while the flour of defatted pre-pupal showed levels around 58, 3% crude protein and 6.6% fat. The pre-pupal meal aminogram showed levels of essential amino acids above traditionally used protein sources, such as meat-and-bone meal, poultry viscera meal and soybean meal. The analysis of the fatty acids obtained from the larva brought high levels of lauric and palmitic acid, corroborating with the literature. This work has provided a good indication that the mass production of black soldier fly for degradation of organic waste and subsequent use as an animal protein is an excellent sustainable alternative to problems related to organic solid waste and alternative sources of protein. / Entre os problemas que o crescimento da população mundial explosiva traz ao meio ambiente, este trabalho propõe uma possível solução para mitigar dois deles: descarte de resíduos sólidos orgânicos e pressão sobre o fornecimento atual de proteína alimentar. Esta solução baseia-se na produção em massa de larvas de Hermetia illucens (L., 1758) (Diptera: Stratiomyidae), também conhecida como mosca soldado negra, para degradar resíduos sólidos orgânicos e também como fonte alternativa de proteína animal. A otimização desta solução reside na compreensão da biologia do inseto para descobrir a relação da qualidade e quantidades ideais de matéria orgânica para garantir sua degradação e aumentar o crescimento das larvas. Finalmente, conhecer a qualidade da proteína larval é fundamental para encontrar seu mercado consumidor. Para entender essas questões, foram realizadas experiências e análises quantitativas. Os resíduos sólidos orgânicos utilizados como alimento para as larvas mostraram na sua composição centesimal 66% de umidade, 63,8% de carboidratos, 13,8% de proteína bruta e 18,1% de gordura. Para a produção em massa de larvas de Hermetia illucens, o estudo procurou entender seu ciclo de vida no bioma amazônico em comparação com a literatura existente. Em relação à biologia das larvas, foi possível observar uma taxa de oviposição em torno de 1002 ovos / fêmea. O tempo de acasalamento leva cerca de 15 minutos. Observou-se um ciclo de cerca de 36 dias desde a oviposição até a idade adulta. Os ovos eclodiram em uma média de 74,75 horas. O tempo de desenvolvimento da larva do ovo para a fase pré-pupae levou cerca de 21 dias e o tempo de pupação 17,5 dias. 74,48% dos ovos atingem o estágio adulto com uma proporção sexual de 3 machos para 1 fêmea. No que diz respeito ao fornecimento da quantidade ótima de alimentos, concluiu-se que, para cada 100 mg de ovos de Hermetia illucens, o suprimento de 2000g de resíduo de restaurante em 2 frações de 1000 g com intervalo de alimentação de 5 dias atingiu o melhor potencial de redução de matéria orgânica. Observou-se uma redução de peso de resíduos sólidos orgânicos de 83,75% com uma taxa de conversão de resíduos sólidos em biomassa proteica de 23,2%. A pré-pupa como matéria-prima para a produção de proteínas animais apresentou 40% de proteína bruta e 32,6% de gordura, enquanto a farinha de pré-pupa desengordurada apresentou níveis em torno de 58,3% de proteína bruta e 6,6% de gordura. O aminograma da farinha de pré-pupa mostrou níveis de aminoácidos essenciais acima das fontes de proteína tradicionalmente utilizadas, como farinha de carne e osso, farinha de vísceras de aves e farelo de soja. A análise dos ácidos gordurosos obtidos a partir da larva trouxe altos níveis de ácido láurico e palmítico, corroborando com a literatura. Este trabalho forneceu uma boa indicação de que a produção em massa da mosca soldado negra para a degradação de resíduos orgânicos e posterior utilização como proteína animal é uma excelente alternativa sustentável aos problemas relacionados aos resíduos orgânicos sólidos e fontes alternativas de proteína.

Black Soldier Fly Larvae (BSFL) composting is a treatment procedure that generates two products: frass compost and BSFL. These products have different uses. BSFL can be used as animal feed and frass compost as soil fertiliser, and they need to be separated after the composting procedure has been finished. A separator of the vibrating sieve type, currently in use by a research group at SLU for separating BSFL from the frass compost generated by the composting process, was tested and evaluated in terms of fraction purity and separation time. It was found that the fraction purity was over 90% for the BSFL fraction, and that the separation time was approximately 1-1.5 minutes for 4 kg of material. Hotspots for improvement were identified and a new vibrating sieve was designed and built with the aim of providing flexibility for improving fraction purity, separation time and operator ergonomy for the end-customer. A CAD model of the new vibrating sieve was designed in SolidWorks with a complete set of technical drawings and purchasing components. These drawings were sent out for manufacturing at a workshop and the new vibrating sieve was built and tested. The new vibrating sieve was functioning properly, but its optimal mechanical settings need further research and testing. This separator design could be emulated by others, and also improved, and this research could be used for improving the process outcome for future BSFL separation processes.

Abstract The management and handling of organic waste in primarily low-income developing cities, due to high urbanization has become a global problem. The main form of waste handling in these cities are landfilling, due to it being the cheapest and easiest option. Not only are nutrients wasted, but landfills of mixed waste create strong emission of greenhouse gasses and a leachate that affects groundwater quality. This major problem in addition to a higher demand for food due to the increase of the world population leaves us two problems to solve; to handle waste and produce food. The Black Soldier Fly (Hermetia illucens) and its larvae can solve both of these problems in one. The organic waste is fed to the fly larvae and the larvae produce biomass (larval weight) and a residue similar to un-mature compost. The larvae can then be fed live to animals or made into pellets to feed animals such as fish, and the residue can be used as fertilizer. However, in order for the larvae to be able to be extruded into pellets or fed live to ex. hens, an optimal protein and fat content of the larvae is needed. A higher protein and lower fat content of the larvae is optimal for both pellet production and to feed to animals. It has previously been found that an inclusion of mussels can lower the fat content and increase the protein content of the larvae. In this article, different amounts of mussels are included in the larvae diet apart from bread and cabbage to see if it has an effect on the compost process efficiency. The experiment is done by larvae being added to different ratios of bread: cabbage: mussels. The larvae are then harvested and process efficiency parameters; BCE (biomass conversion efficiency), survival percentage, material reduction and respiration are calculated. It was found that mussel inclusion was equivalent to a higher feeding load per larva. The BCE of the treatment with a higher feeding load and the treatment with the highest mussel inclusion were the same, meaning that mussel inclusion is equivalent to a diet with high amounts of carbohydrates. Mussel inclusion in the larva diet seems to be beneficial from a BCE standpoint and suits as a substrate as pH and survival was not negatively affected by the mussels.

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