{"paper_id":"0bd13d35-e120-4ce8-8886-6ebd229bd4cf","body_text":"Physical-Chemical Composition Analysis of insect frass from different species produced through the bioconversion of agro-industrial waste | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Physical-Chemical Composition Analysis of insect frass from different species produced through the bioconversion of agro-industrial waste Rafael Martins da Silva, Andreas Köhler, Daniela Costa, Diego Prado de Vargas, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7509675/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The objective of the present study was to develop organic fertilizers through the bioconversion of agro-industrial waste, utilizing various insect species, with the intent of evaluating both the quantity and quality of the resulting products. To conduct the research, a controlled rearing of the insect species Tenebrio molitor , Zophobas morio , and Hermetia illucens was established under laboratory conditions, incorporating diverse substrates and varying concentrations to assess the composition of the frass produced. The physicochemical characterization included analyses of moisture content, total organic carbon, pH, cation exchange capacity, and micronutrient determination via atomic absorption spectrometry, in compliance with current regulations for organic fertilizers. The primary plant nutrients, such as total carbon (36-44%), nitrogen (2.7-5%), phosphorus (2.5-6%), and potassium (1.7-3.72%), exhibited variation depending on the substrate and insect species used, with macro and micronutrient values in alignment with the requirements set forth by Brazilian and European Union legislation. Furthermore, no contaminants were detected in any of the samples analyzed. It can be concluded that the frass produced by T. molitor , Z. morio , and H. illucens demonstrates significant potential as an organic fertilizer, as its concentrations of nitrogen, phosphorus, and potassium are comparable to those found in other organic fertilizers on the market. This suggests its potential to fully replace both traditional commercial fertilizers and organomineral fertilizers, thereby contributing to a reduction in the carbon footprint associated with fertilizer production. Residues Fertilizer Entomology Regulation 1 Introduction Current projections indicate that by the year 2050, global waste production is anticipated to reach approximately 3.4 billion tons, with a predicted 44% of this being biodegradable organic matter. It is estimated that the majority of this waste will be generated in underdeveloped or developing countries (Lopes et al., 2022). In addition to the waste of organic material, which could be reused in various applications and technologies, the final disposal of this waste is carried out in landfills and open dumps (approximately 70%), having a serious impact on the environment due to the release of greenhouse gases, leachate with toxic compounds and the formation of environments conducive to vectors of pathogenic organisms (Koda et al. 2017; Lopes et al. 2022). The utilization of edible insects in bioconversion represents a promising environmental technological method. This approach has the potential to reduce the volume of biodegradable organic waste, thereby eliminating the need for final disposal in landfills. Additionally, it can mitigate greenhouse gas emissions and environmental contamination by toxic leachate (Gold et al. 2018; Chiam et al. 2021). Furthermore, the bioconversion of insects such as the Black Soldier Fly (Hermetia illucens [Diptera: Stratiomyidae]), Tenebrio molitor (Coleoptera: Tenebrionidae), and Zophobas morio (Coleoptera: Tenebrionidae) has been demonstrated to be a viable solution. The larvae of BSF have been shown to reduce approximately 45% to 80% of the organic waste available for bioconversion within a relatively short time frame of 2 to 5 weeks (Chiam et al. 2021; Terfa 2022; Jenkins et al. 2023). Furthermore, the employment of BSF in bioconversion or waste reduction processes exhibits enhanced efficiency in comparison to vermicomposting processes executed by worm species such as Eisenia foetida . This superiority can be attributed to the capacity of BSF to consume a more diverse range of waste, obviating the necessity for desalination processes of organic materials, thereby reducing operational costs and enhancing the overall practicality and viability of the process (Xiao et al. 2018). Finally, a notable benefit of employing BSF in the context of organic matter is its capacity to impede the proliferation of other fly species, which can act as vectors for pathogenic organisms (Chiam et al. 2021). Given the propensity of the species H. illucens , T. molitor , and Z. morio to reproduce more efficiently in tropical and subtropical regions, the implementation of bioconversion technology employing these insects presents a significant opportunity for innovation, particularly in developing countries (Barragan-Fonseca et al. 2017; Fuhrmann et al. 2022). Another salient aspect of the bioconversion of agro-industrial waste pertains to the frass that is produced in the process (residual substrate from bioconversion or organic fertilizer). This frass is promoted by insects of various species, including T. molitor , Z. morio and H. illucens , which have the capacity to provide nutrients necessary for the development of various crops, both organically and conventionally (Setti et al. 2019; Terfa 2021). Research has been conducted to obtain simple or compound organic substrates and fertilizers from agro-industrial waste. The objective of this research is to replace synthetic and inorganic fertilizers, which have a high impact on the environment and human health, from the manufacturing process to their use. (Ronga et al. 2016; Setti et al. 2019; Poveda et al. 2019; Quilliam et al. 2020; Blakstad et al. 2023). It is crucial to acknowledge that agricultural practices reliant upon chemical fertilizers neglect the imperative of sustaining optimal levels of organic matter within the soil. This oversight directly impinges upon the preservation of the soil's physical and chemical properties, thereby exerting a detrimental influence on its capacity for productivity. Moreover, the extensive utilization of mineral fertilizers, in conjunction with their management, poses a significant threat to the sustainability of agriculture. This is due to the fact that they contribute to the progressive deterioration of soils, which can result in depletion, salinization, or desertification (Przemieniecki et al. 2021; Fuertes-Mendizábal et al. 2023). Among the plant macronutrients, nitrogen stands out as a particularly demanding nutrient, and one that poses significant challenges in terms of supplementation. This is due in large part to the significant loss of nitrogen that occurs during the production and application of urea-based fertilizers. The loss of nitrogen in this process can range from 40% to 70%, which has far-reaching environmental consequences. The contamination of groundwater with leached nitrates is a primary concern, as it directly impacts water quality and availability. Additionally, the atmosphere becomes exposed to the release of greenhouse gases, such as NH­ 3 volatilization and N 2 O, which contribute to both local and global climate change. The production of ammonia-based fertilizers is a significant contributor to environmental degradation, with the production of 1,000 kg of NH 3 resulting in the generation of 2,000 kg of CO 2 eq. The process is highly dependent on natural gas, as outlined in the works of Li et al. (2021), Barbi et al. (2022), Jenkins et al. (2023). The utilization of insect frass in agriculture as an organic fertilizer and phytofortifier has garnered significant attention and interest in recent years. This is due to its nutrient content and compounds that possess the potential to substitute for chemical fertilizers. Additionally, its slow-release mechanism, attributable to its low solubility in water, ensures sustained nutrient availability over extended periods (Blackstad et al. 2023). Entomology has long acknowledged the potential of insect excreta as a source of bioavailable nutrients that are readily assimilated by plants (Poveda et al. 2019). A limited number of studies have examined the utilization or efficacy of the organic compost produced in the bioconversion process as an organic fertilizer or soil conditioner. The findings from these studies are often inconsistent (Chiam et al. 2021). It is also important to note that the components and parameters of the frass are substrate-dependent, which complicates the comparison of results across studies due to variations in substrate source and type (Poveda et al. 2019). The edible insect species T. molitor produces frass, which has the potential to provide valuable nutrients for plant growth and development when used as a fertilizer or soil conditioner. The NPK balance of frass is typically 3.5-1.5-1.5 (Liu et al. 2003; Poveda et al. 2019). However, it is crucial to acknowledge that both the NPK values and the composition of other elements are contingent on the substrate (organic matter) provided during the larval development of T. molitor . Recent studies on agricultural waste have demonstrated that this insect species possesses the capacity to survive and biodegrade organic matter, including lignin, thereby generating high-value-added products, such as larval biomass and frass. However, millions of tons of excrement waste generated are not treated properly by T. molitor breeding companies, reinforcing that their correct use could significantly contribute to increasing rural incomes by using this frass as an easily available organic fertilizer (He et al. 2021). A notable advantage of T. molitor and Z. morio over other insect larvae, such as BSF used for food waste bioconversion processes, is that T. molitor adults or Z. morio beetles have fused wings/elbows and do not fly, making their biocontainment significantly easier and reducing the risk of their escape causing problems for the local ecosystem (Gan et al. 2021). Frass from the T. molitor rearing process is a mixture of larval feces, undigested organic waste and discarded exoskeletons (Fuertes-Mendizábal et al. 2023). It has been demonstrated that this by-product has the capacity to increase the tolerance of different plants to abiotic stress parameters and resistance to biotic stress from different sources (Gebremikael et al. 2022). This behaviour is linked to the direct contribution of nutrients, organic carbon and, among other parameters, the presence of bioavailable chitin molecules that stimulate the immune system, as well as plant growth-promoting microorganisms (Houben et al. 2020; Barrágan-Fonseca et al. 2022; Blakstad et al. 2023; Hénault-Ethier et al. 2023). It is important to note that the chitin in the exoskeleton of T. molitor needs to be degraded by soil organisms in order to become bioavailable to the plant. The conventional composting process, which can take 8 to 24 weeks, is contrasted with the accelerated composting process promoted by BSF, which takes only 5 weeks to convert organic waste into stable organic fertilizer (Anyega et al. 2021). The benefits of organic fertilizer include cost savings, environmental friendliness, and the indirect promotion of sustainable agriculture by reducing waste of organic matter (Janah et al. 2023). Furthermore, the Frass produced through bioconversion promoted by BSF accounts for 30% to 50% of the initial weight of the food substrate supplied, with this amount depending on the type and quality of the waste (Amrul et al. 2022). Finally, it is imperative to acknowledge the pivotal role of frass in stimulating the growth of microorganisms present in the soil, accelerating the rate of decomposition, and promoting soil respiration, nitrogen immobilization, and mineralization (van Zande et al. 2023). Moreover, extant studies have demonstrated that frass enhances plant growth, quantity, nutrient absorption, N use efficiency, and disease suppression in various crops (Quilliam et al. 2020; Anyega et al. 2021). It is noteworthy that organic fertilizers also contain nitrifying and denitrifying bacteria, which play a crucial role in the nitrogen cycle, thereby facilitating plant nitrogen uptake (Terfa 2021). While some farmers have reported the beneficial effects of excrement on plants, there are gaps in the information on the fertilizing capacity of excrement produced by insects to improve or condition soil fertility and, ultimately, plant growth. Furthermore, as emphasized by most researchers, the fertilizing potential of frass requires more research and data before the industry can expand. This is also relevant due to the search for economic and ecological alternatives to conventional mineral fertilizers, whose production depends on fossil fuels and finite resources (Poveda et al. 2019; Houben et al. 2020; He et al. 2021). Consequently, this study was undertaken to produce organic fertilizers from the bioconversion of agro-industrial waste promoted by different species of insects, for subsequent analysis of their quantity and quality. 2 Materials and Methods 2.1 Rearing Hermetia illucens (Black Soldier Fly - BSF) larvae and obtaining frass. The insects were reared in the Entomology Laboratory at the University of Santa Cruz do Sul (UNISC) in air-conditioned rooms with a temperature of 26 ± 2°C, relative humidity of 60 ± 5%, and a 12-hour photoperiod (adult stage). The larval stage was maintained in air-conditioned rooms with temperatures of 28°C ± 2°C and relative humidity of 60%. As these insects are photosensitive, they were kept in the absence of a photoperiod (dark). The adults were kept in breeding cages made from white crystal organza fabric (90 cm high, 40 cm deep, and 30 cm wide) for continuous oviposition. These cages were maintained with a layer of attractant (fermented starter chick feed) and grouped strips of wood (eucalyptus, 5 cm x 20 cm x 2 cm), which served as oviposition sites.Subsequent to the eggs being laid (an average of 2 days), the wooden strips with the eggs were extracted and placed in trays with feed for the initial larvae to hatch. It is noteworthy that the oviposition wood was replenished every two days in all the rearing cages. During the growth phase, the larvae were placed in containers with different substrates, including residual whole wheat flour (E. Kuehniella rearing), orange waste, apple and banana waste (fruit mix), in different proportions (Table 1), and fed ad libitum. In the final stage (pre-pupa stage), the last-stage larvae were separated from the substrate using sieving processes to dislodge the larvae from the frass (organic fertilizer) for subsequent analysis and utilization. 2.2 Breeding Zophobas Morio and Tenebrio Molitor and obtaining frass The rearing of both species was conducted at the Entomology Laboratory of the University of Santa Cruz do Sul (UNISC) in air-conditioned rooms with a temperature of 28 ± 2 °C, relative humidity of 60 ± 5%, and a 12-hour photoperiod. The adults were maintained in plastic breeding boxes (46.7 cm x 32.3 cm x 17.9 cm) for the purpose of continuous oviposition. These boxes were maintained with a layer of substrate (wheat bran) and a water source (fruit and vegetables). Subsequent to egg-laying and the emergence of the first larvae, the boxes were sieved to separate the adults, eggs, and first-stage larvae. During the growth phase, the larvae of both species were placed in containers with substrate (wheat bran) and a water source, and provided with an unlimited supply of food. The abiotic parameters utilized in the maintenance of the adults were employed in this instance as well. In the final stage, the last instar larvae were subjected to a sieving process to separate them from the frass (organic fertilizer), which was subsequently analyzed for its composition and quality. Table 1 - Rearing parameters for the different species of bioconverter insects Espécie Substrato Temperatura Fotoperíodo Z. morio (ZM-F) Farelo de Trigo, Laranja e Chuchu 28 °C ± 2°C 12 h T. molitor (TM-F) Farelo de Trigo, Laranja e Chuchu 28 °C ± 2°C 12h H. illucens (BSF-80O) 80 % farinha trigo integral - 20 % Laranja 26 °C ± 2°C Escuro H. illucens (BSF-10O) 90 % farinha trigo integral - 10 % Laranja 26 °C ± 2°C Escuro H. illucens (BSF-90FM) 10 % farinha trigo integral - 90 % Mix de Frutas 26 °C ± 2°C Escuro H. illucens (BSF-10FM) 90 % farinha trigo integral - 10 % Mix de Frutas 26 °C ± 2°C Escuro 2.3 Compositional analysis 2.3.1 Preparation The sample was prepared by manually homogenizing the entire sample and reducing it by quartering until a quantity of approximately 300 g was obtained.A portion of 30 to 40 g was set aside for determining the pH of the sample in natura, and this amount was then divided into two equal fractions. One portion was allocated for granulometric analysis, employing the method delineated in the Manual of Official Analytical Methods for Fertilizers and Correctives of the Ministry of Agriculture and Livestock. The other portion was designated for chemical analysis. Subsequently, the largest portion of the sample was weighed in natura and transferred to a soil drying oven (TE-394/5 - Tecnal) at a temperature of 65 ± 5°C until it attained a constant weight. The fraction intended for chemical analysis was subjected to comminution and sieved through a mesh opening of 500 µm. 2.3.2 Moisture, total organic carbon, pH and Cation Exchange Capacity analyses The quantification of organic carbon was executed through the implementation of the potassium dichromate volumetric method, which entailed the wet oxidation of the organic carbon present within the sample with excess potassium dichromate and concentrated sulfuric acid, accompanied by external heating.The moisture content was determined employing the 65°C moisture determination method (U65). This method involved the calculation of the percentage of moisture in the sample at 65°C, utilizing the data (G1 and G2), according to the following expression: where: G1 = mass of the \"in natura\" sample, and G2 = mass of the sample dried at 65°C (m/m).The pH was subsequently measured using the pH determination technique, which consists of suspending the sample in a 0.01 mol L -1 CaCl 2 solution and measuring the pH potentiometrically. The total nitrogen was determined by means of the Raney alloy macro-method, which consists of the ammonification of all non-ammoniacal forms of nitrogen, including organic forms. This was followed by the alkaline distillation of ammonia, which was treated with an excess amount of boric acid. The measurement of total phosphorus involved the dissolution of the phosphorus in the sample through strong acid extraction, followed by the precipitation of the orthophosphate ion as quinoline phosphomolybdate. This was then filtered, dried, and weighed. Water-soluble potassium was determined by hot extraction of water-soluble potassium and quantification by volumetry using flame photometry. Silicon was quantified using the ammonium molybdate spectrophotometric method after extraction with cold hydrochloric and hydrofluoric acids. The CTC/C ratio was determined by calculating the ratio between the values found for cation exchange capacity (CTC), in mmolc kg-1, and organic carbon, in mass percentage, both referred to the sample on a dry basis. The unit of measure for the CTC/C ratio is 𝑚𝑚𝑜𝑙𝑐.10 -1 g 𝑑𝑒 C. It is imperative to acknowledge that this ratio serves as a metric for evaluating the degree of maturity and quality of organic fertilizers. 2.3.1.1 Micronutrient analysis An analysis was conducted on the concentrations of phosphorus (P), potassium (K), sodium (Na), copper (Cu), calcium (Ca), iron (Fe), magnesium (Mg), zinc (Zn), and manganese (Mn). The atomic absorption spectrometry method was employed in accordance with the Manual of Official Analytical Methods for Fertilizers and Correctives of the Brazilian Ministry of Agriculture, Livestock and Supply. 3 Results The results demonstrate that the total nitrogen value (N total) varies statistically according to the diet administered, the concentration of waste, and the species of insect responsible for bioconversion, as well as the statistical difference between the insects and the control ( Ephestia kuehniella residual meal). The BSF diets with the lowest residue concentrations, BSF-10FM and BSF-10O, exhibited the highest nitrogen concentrations (4.93% and 5.07%, respectively), with no statistical difference observed. Additionally, diets with a higher amount of waste for BSF, T. molitor , Z. morio , and the control demonstrated no statistical difference, with an approximate result of 3.20%. It is noteworthy that all diets met the minimum levels stipulated by the Ministry of Agriculture and Livestock (MAPA) for organic fertilizers, with the frass content aligning with the established criteria. With regard to phosphorus, the control showed the highest result (≅ 6%), followed by BSF-10FM and BSF-10O (≅ 5.4% and ≅ 4.91%), which were statistically equal. The remaining formulations and species exhibited statistically equivalent lower results and divergent higher values, with the exception of BSF-90FM, which demonstrated the lowest value (≅ 2.58%) and was statistically divergent from all species and formulations. Potassium values were found to be comparatively low in relation to nitrogen (N) and phosphorus (P), with the exception of BSF-90FM, which exhibited the highest result (approximately 3.72%), statistically different from the others. This was followed by the control and BSF-10FM (approximately 2.43% and 2.77%, respectively). The remaining species and formulations demonstrated statistically equal values. Micronutrients, including calcium, magnesium, molybdenum, silicon, iron, and sulfur, exhibited comparable concentrations across all species and formulations, with minimal numerical variations. However, micronutrients such as boron, copper, cobalt, nickel, and zinc demonstrated statistical differences between species and formulations. With regard to boron, BSF-80O exhibited the highest levels (≅ 615.05 mg.kg -1 ), which were statistically different from the others, followed by the formulations with 10% residue BSF-10FM, BSF-10O, and control (average of ≅ 455 mg.kg- 1 ). Among the micronutrients that demonstrated statistical differences was copper, where the highest values were obtained in the control and BSF-10FM (average of ≅ 8.5 mg.kg -1 ). Similarly, manganese exhibited higher levels in the TM-F and ZM-F species of tenebrios (417.56 mg.kg -1 and 318.99 mg.kg -1 ). The pH exhibited statistical differences only in BSF-80O and the control (average of ≅ 7.3) in comparison to the other species and formulations. Total carbon levels in all the formulations demonstrated levels that exceeded the minimum requirement (15% MAPA reference), with the lowest levels observed in BSF-10O and the control (38.61% and 36.22%, respectively). The cation exchange capacity (CEC) exhibited higher values in the BSF-10FM and BSF-90FM samples (706.32 mmol.kg -1 and 594.76 mmol.kg -1 ), respectively, both of which incorporated multiple fruits in their formulation (banana and apple). Table 1 - Composition of the frass produced by each species (MAPA) Parameters Min. Levels TM-F ZM-F BSF-10 FM BSF-90FM BSF-10O BSF-80O Control Granulometry Classificação Powder Powder Bran Bran Bran Bran Bran Max humidity 40.00% 6.79a 9.51b 7.67a 7.07a 13.69d 4.83c 19.02e pH As stated 5.78a 5.75a 6.41a 7.57b 6.42a 6a 7.22b C min. 15.00% 44.23a 42.94b 41.72b 43.07ab 38.61d 46.56c 36.22e Total N min. 0.50% 3.35a 2.71a 4.93b 3.16a 5.07b 2.94a 3.8ac CTC (mmol/Kg) As stated 316.91a 312.19a 706.32b 594.76c 414.29d 380.68d 408.95d CTC/C As stated 7.17a 7.27a 16.93b 13.81c 10.73d 8.18a 11.29d Nitrogen (N) 1.000% 3.35a 2.71a 4.93b 3.16a 5.07b 2.94a 3.8a P 2 O 5 1.000% 4.03a 4.57a 5.4b 2.58c 4.91bad 3.5a 6.03be K 2 O 1.000% 1.86a 1.81a 2.77b 3.72c 1.73a 1.9ad 2.43bd Ca 1.000% 2.83a 2.8a 2.85a 2.7a 2.61a 2.97a 2.67a Mg (%) 1.000% 0.69a 0.71a 0.79a 0.6a 0.64a 0.41b 0.74a S (%) 1.000% 0.95a 0.24b 0.35b 0.23b 0.28b 0.42b 0.38b B (mg/Kg) 0.010% 417.56a 318.99b 454.31c 341.05b 456.37c 615.05d 457.93c Co (mg/Kg) 0.005% 5.37a 5.21a 6.65b 9.37c 6.21bd 5.48a 7b Cu (%) 0.020% 12.93a 15.35b 18.5c 15.05d 14.64e 13.2a 18.72 Fe (%) 0.020% 0.02a 0.03a 0.03a 0.03a 0.03a 0.02a 0.03a Mn (mg/Kg) 0.020% 314.73a 346.18b 273.62c 121.84d 120.91d 51.28e 255.26c Mo (mg/Kg) 0.005% <15a <15a <15a <15a <15a <15a <15a Ni (mg/Kg) 0.005% <2a 2.4b 3c 3c 2.2b 2.5b <2a Se (mg/Kg) 0.003% <1a <1a <1a <1a <1a <1a <1a Si (%) 0.050% 0.03a 0.06a 0.06a 0.06a 0.03a 0.03a 0.02a Zn (mg/Kg) 0.100% 110.42a 118.06a 129.81b 68.06c 100e 47.74d 122.43ab To comprehensively assess the various organic fertilizers derived from bioconversion, it was imperative to analyze the contaminants, as mandated by MAPA, Brazil's regulatory authority. As delineated in Table 2, the frass from all species and dietary regimens exhibited negligible levels of contamination by heavy metals, including arsenic, cadmium, lead, hexavalent chromium, and mercury. With regard to nickel, it was detected above the minimum analysis limits in the organic fertilizers, with the exception of the TM-F frass (<2 mg.kg -1 ), but all were well below the contamination levels (70 mg.kg -1 ). It is imperative to note that the maximum tolerance level for thermotolerant coliforms is 1000 NMP.g -1 . However, it is noteworthy that with the exception of the TM-F sample (450 NMP.g -1 ), all the samples were found to be free of these. Furthermore, the tests revealed the absence of helminth eggs and Salmonella sp. in all the frass samples across all species and diets. Finally, it is noteworthy that all the organic fertilizer samples were found to be free from contamination, thereby meeting the conditions for use as stipulated by Brazilian legislation in accordance with MAPA standards and legislation from other countries. Table 2 - Limits of contaminants allowed by Brazilian legislation (MAPA) Contaminants Allowed Parameters TM-F ZM-F BSF-80O BSF-10O BSF-90 FM BSF-10 FM As 20 mg.kg -1 <1 <1 <1 <1 <1 <1 Cd 3 mg.kg -1 <0.2 <0.2 <0.2 <0.2 <0.2 <0.2 Pb 150 mg.kg -1 <2 <2 <2 <2 <2 <2 Cr-VI 2 mg.kg -1 <0.4 <0.4 <0.4 <0.4 <0.4 <0.4 Hg 1 mg.kg -1 <0.2 <0.2 <0.2 <0.2 <0.2 <0.2 Ni 70 mg.kg -1 <2.0 2.4 2.5 2.2 3 3 Se 80 mg.kg -1 <1 <1 <1 <1 <1 <1 Helmints Eggs 1/4g Negative Negative Negative Negative Negative Negative TtC 1000 450 <180 <180 <180 140 <180 Salmonella sp. Absence Absence Absence Absence Absence Absence Absence 4 Discussion The waste degradation efficiency and bioconversion periods of insect species vary, as does the amount of frass fertilizer available. In general, the larvae of H. illucens exhibit a high waste degradation efficiency (55-80%) (Lalander et al. 2015; Beesigaukama et al. 2022; da Costa e Silva et al. 2024), a relatively brief bioconversion time, and consequently, they produce higher quantities of organic fertilizer in comparison to other insect species. Conversely, T. molitor and Z. morio require 4 to 6 weeks to convert organic waste into fertilizer, yielding comparatively lower quantities of frass. Lignocellulosic materials impose a significant stressor on the digestive system of the BSF, resulting in a diminished concentration of organic matter in frass (Manurung et al. 2016). The degradation and conversion of lignocelluloses by BSF is considered viable but inefficient (Liu et al. 2021a). The characteristics of insect frass, including its composition and quality, exhibit variability due to the substrate provided to the larvae during the rearing process and subsequent treatment. This variability may be attributed to legislative and/or health-related recommendations (Klammsteiner et al. 2020; Lomonaco et al. 2024). The average concentration of carbon (C) has been documented to range from 35% to 40% (Chiam et al. 2021; Watson et al. 2021; Song et al. 2021; Lopes et al. 2022), which is in accordance with the findings of this study, which identified concentrations in all species and different diets that varied between 38% and 48%, inferring that the larvae have the ability to concentrate organic matter and accumulate nutrients, even if the frass is the product of their excretion plus the remains of the substrate they are fed, since the control had lower amounts of C (≅ 36%). The concentrations of nitrogen (N), phosphorus (P), and potassium (K) found in these species are comparable to those found in farmyard manure, particularly poultry manure (Poveda et al. 2019), thereby substantiating their considerable fertilizer potential. In contrast to conventional mineral fertilizer, frass also contains trace amounts of micronutrients (e.g., copper [Cu], zinc [Zn], manganese [Mn], and iron) that can be particularly advantageous for plant growth. The carbon (C) fraction in frass is comparable to that of pig and poultry manure (Rothé et al. 2019; Liu et al. 2019), with a substantial soluble C fraction, which may be associated with the efficient digestion of cellulose and lignin compounds by T. molitor , Z. morio , and BSF larvae (Houben et al. 2020). It is noteworthy that all types of frass from diverse species and substrates exhibited the majority of essential trace elements for plant growth, with the exception of Ni. The nitrogen (N) concentrations did not exhibit significant variation across species or dietary treatments (approximately 3%), suggesting that the diets did not substantially influence the nitrogen content in frass. These findings align with those reported by Menino et al. (2021), Watson et al. (2021), Anyega et al. (2021), Green (2023), and Lomonaco et al. (2024) (Table 3). However, in the present study, higher N concentrations were observed in BSF diets supplemented with only 10% waste, which contained elevated levels of starch and carbohydrates due to the inclusion of flour. Previous studies have suggested that the incorporation of protein into diets can enhance nitrogen concentrations in frass (Setti et al. 2019). Nevertheless, it is well established that larvae generally exhibit suboptimal development on diets rich in protein but deficient in carbohydrates, which in turn leads to insufficient frass production due to nutrient limitations (da Costa e Silva et al. 2024). Furthermore, Sarpong et al. (2019) reported that diets characterized by high protein and sugar/starch content, coupled with low fiber concentrations, promote the accumulation of nutrients in frass. Table 3 - Results of recent research on biofertilizers produced or BSF bioconversion. (Chiam et al., 2021) BSF - Fresh Okara (Menino et al., 2021) BSF - Potato and Onion (Watson et al., 2021) T.molitor - Insectus Mealworm Grow (Gärttling et al., 2020) BSF - Maize (Song et al., 2021) BSF – Okara and wheat bran (Anyega et al., 2021) BSF – Brewer´s spent grains (Kawasaki et al., 2020) BSF – Household organic waste C 37,07% 38,7% 35,7% 41,8% 37,1% 38,6% 35,8% N 5,15% 2,8% 2,8% 3,3% 4,78% 3,6% 2,2% S AD - - - 0,5% - - K 1,93% 3,3% 2,3% 2,4% 0,985% 0,3% 0,7% P 0,293% 1,5% 1,4% 3,4% 1,009% 0,5% 0,5% Ca 16,7 mg.kg -1 15 g.kg -1 - 4 g.kg -1 1341 mg.kg -1 9,7 g.kg -1 10 g.kg -1 Mg 10,5 mg.kg -1 7 g.kg -1 0,3 g.kg -1 10 g.kg -1 11,78 mg.kg -1 1 g.kg -1 0,9 g.kg -1 B 4,77 mg.kg -1 - - - 2,77 mg.kg -1 - - Fe 3,69% 896 mg.kg -1 15 mg.kg -1 - 26,55 mg.kg -1 310 mg.kg -1 240 mg.kg -1 Mn 0,19% 149 mg.kg -1 19,4 mg.kg -1 - 4,21 mg.kg -1 109 mg.kg -1 10 mg.kg -1 Cu 0,86% 19 mg.kg -1 8,9 mg.kg -1 - 2,21 mg.kg -1 25 mg.kg -1 10 mg.kg -1 Mo 0,26% - - - 2,77 mg.kg -1 - - Zn 1,73%) 137 mg.kg -1 15 mg.kg -1 - 0,10 mg.kg -1 182 mg.kg -1 10 mg.kg -1 Ni 0,11% - - - - - - As - - - - - - - Cd - - - - - - - Pb - - - - - - - Cr - - - - - - - C/N 13,8 16 12,6 7,76 10,7 16,6 In general, the predominant form of nitrogen in black soldier fly (BSF) frass is ammonium nitrogen (NH₄⁺-N), while nitrate nitrogen (NO₃⁻-N) concentrations tend to be lower. This is likely due to the fact that BSF frass is processed solely through the larval digestive system, in contrast to fertilizers derived from animals such as poultry and cattle, where the substrate undergoes microbial fermentation during digestion. Moreover, insect frass, unlike conventional livestock manure, represents a novel fertilizer with a lower potential for NO₃⁻-N accumulation in plants, thereby reducing the risk of nitrate-induced toxicity (Kawasaki et al., 2020). Beyond nitrogen content, the nutritional advantages of insect frass extend to other essential elements, as evidenced by increased uptake of phosphorus (P), potassium (K), magnesium (Mg), and sulfur (S) in plants fertilized with frass compared to those treated with conventional organic fertilizers. Additionally, the elevated phosphorus concentrations in frass may enhance nitrogen assimilation in plants, as phosphorus plays a crucial role in energy transfer processes (Fageria 2001; Kawasaki et al. 2020). Although the total carbon content remained consistent across all frass samples, the C/N ratio varied among them, with values of 13.20 for TM-F, 15.84 for TM-Z, 8.46 for BSF-10FM, 13.63 for BSF-90FM, 7.61 for BSF-10O, and 15.84 for BSF-80O. A C/N ratio exceeding 20 presents a risk of nitrogen immobilization in the soil; however, the nitrogen content in frass can benefit plants with efficient nitrogen utilization, which is largely influenced by their rhizobiome (Klammsteiner et al. 2020; Dzepe et al. 2022). Furthermore, insect larvae undergo six developmental instars, during which they periodically shed their exoskeleton. This exoskeleton is primarily composed of chitin, a polymer of N-acetylglucosamine (C 8 H 13 O 5 N) n , which affects the C/N ratio. Additionally, chitin degradation can yield chitosan, a compound with plant growth-promoting properties that also enhances resistance to pathogens (Sharp 2013; Klammsteiner et al. 2020; Nayak et al. 2020; Fuertes-Mendizábal et al. 2023). Supporting this premise, as well as the findings of the present study, Anyega et al. (2020) suggests that the increased growth rates and yields observed in vegetables cultivated with BSF frass may be attributed to its advanced maturity and stability, as evidenced by a lower C/N ratio. The pH variation observed in the BSF frass samples was minimal (6.00–7.57), exhibiting a slight tendency toward alkalinity. In contrast, the pH values of TM-F and ZM-F remained nearly constant (5.75–5.78), indicating a slightly more acidic nature. The average pH value reported in the literature is 7.46, which is also mildly alkaline and, as observed in this study, represents one of the least variable parameters. Furthermore, it is well established that BSF, in its organic form, actively alters the pH of the substrate toward alkaline levels, thereby supporting the findings of this study (Gärttling & Schulz 2022). Additionally, the higher nutrient concentrations and lower C/N ratio associated with organic insect-derived fertilizers underscore the remarkable efficiency of BSF, T. molitor , and Z. morio larvae in nutrient recycling (Lalander et al. 2015; Tanga et al. 2022). Compost or biofertilizer maturity refers to the degree of completeness of the bioconversion process, characterized by the absence of phytotoxic compounds as well as animal and plant pathogens that could negatively impact seed germination, plant growth, and soil health. In simple terms, compost maturity signifies its suitability for agricultural application as a fertilizer, assessed through chemical, physical, and biological indicators. During the present study, the C/N ratio ranged from 7 to 16, indicating high-quality and mature compost (Goyal et al. 2015; Houben et al. 2020; da Costa e Silva et al. 2024). Similarly, the pH values of all analyzed species and substrates remained within the range of 6.0 to 7.5, a threshold considered acceptable for mature compost. This finding aligns with the observations of Lomonaco et al. (2024), who reported that Hermetia illucens tends to shift the pH of frass toward more alkaline conditions. Furthermore, the alkalization of frass induced by BSF-driven bioconversion is attributed to the release of ammonium ions (NH₄⁺) and ammonia (Alidadi et al. 2020; Lomonaco et al. 2024). It is important to highlight that the chemical parameters, including macro- and micronutrient concentrations, align with the standards recommended for high-quality fertilizers (Table 1). These fertilizers ensure the release of adequate nutrient levels necessary for optimal plant development. Furthermore, the observed values comply with the regulatory standards established in various countries, including Brazil, the United States, Canada, African nations, and the European Union. The effectiveness of organic fertilizers in vegetable production is closely associated with their nutrient composition, particularly nitrogen (N), and the rate at which these nutrients are released (Cabrera et al. 2005; Fuertes-Mendizábal et al. 2023). The frass derived from Tenebrio molitor , Zophobas morio , and black soldier fly (BSF) exhibits significant potential as an organic fertilizer due to its high concentrations of nitrogen (N), phosphorus (P), and potassium (K), which are comparable to those found in other organic fertilizers, such as raw manure (Houben et al. 2020; Dzepe et al. 2022). This observation aligns with the findings of the present study, as the frass of these insect species contained average N-P-K values of approximately 3% N, 4% P₂O₅, and 1.80% K₂O, respectively. Additionally, Fuertes-Mendizábal et al. (2023) reported a substantial increase in lettuce leaf biomass with the application of only 1% frass, attributed to enhanced nitrogen uptake. Notably, in contrast to conventional organic fertilizers, insect frass is characterized by a high content of labile organic matter measured at approximately 40% in all species and substrates examined in this study and exhibits rapid mineralization kinetics (Chavez a& Uchanski 2021), thereby facilitating the swift availability of nutrients. Studies have demonstrated that the application of Hermetia illucens and Tenebrio molitor frass as fertilizer results in higher yields and improved nutritional quality in various crops, including maize, tomatoes, cabbage, cowpeas, pepper, shallots, and barley, when compared to conventional fertilizers (Tanga et al. 2021; Beesigamukama et al. 2022). Moreover, soil amendment with H. illucens and T. molitor frass has been shown to suppress the proliferation of soil-borne pathogens, enhance soil microbial activity, reduce soil acidity and salinity, improve nitrogen mineralization, and increase nutrient availability, thereby contributing to overall soil quality and promoting plant growth (Houben et al. 2020; Barragán-Fonseca et al. 2022). Additionally, fertilization with black soldier fly (BSF) frass has been observed to enhance potassium (K) uptake by plants, where potassium serves as an enzyme activator essential for maintaining cell membrane integrity and developmental potential (Hellgren et al. 2006; Radzikowska-Kujawska et al. 2023). Under optimal growth conditions, phosphorus (P) absorption is also notably increased (Radzikowska-Kujawska et al. 2023). Furthermore, ongoing research in the literature explores the role of polyphenolic compounds absorbed by plants, particularly those fertilized with insect frass. Among these compounds, ferulic acid has garnered attention due to its efficient absorption from food, its metabolic pathways in the human body, and its potential health-promoting properties (Kwee et al. 2011; Radzikowska-Kujawska et al. 2023). In the European Union, the classification and labeling of frass as an organic fertilizer are regulated under Regulation (EU) 2019/1009, which generally categorizes it as a solid organic fertilizer (PFC 1 (A) (I)). Moreover, this regulation establishes that compound solid organic fertilizers must contain a minimum of 1% total nitrogen (N), phosphorus pentoxide (P₂O₅), or potassium oxide (K₂O), with the combined proportion of these primary nutrients amounting to at least 4% by mass (Houben et al. 2020; IPIFF,2021). In this context, it is important to highlight that the frass derived from all species and diets examined in the present study complies with the regulatory requirements set forth by both the Brazilian Ministry of Agriculture and Livestock and the European Union. Furthermore, the substitution of traditional organic waste treatment methods, such as aerobic composting, with the use of black soldier fly (BSF) larvae has demonstrated a significant reduction in the global warming potential associated with organic waste management, decreasing emissions by approximately 50% (Mertenat et al. 2019). Despite its potential benefits, the application of insect frass as a fertilizer remains an area requiring further investigation. This necessity arises primarily due to the volatilization of nitrogen into potentially toxic forms, which is influenced by soil pH and cation exchange capacity (CEC). Additionally, regulatory frameworks, such as those of the European Union, mandate the standardization of frass through thermal treatment at 70°C, which may further affect its properties and efficacy as a fertilizer. Consequently, the optimization of fertilizer application methods will be a critical consideration in future research and agricultural practices (Kawasaki et al. 2020; Houben et al. 2020; Gärttling & Schulz 2022). 5 Conclusion Insect frass has demonstrated significant potential as a sustainable organic fertilizer, with the capacity to fully replace conventionally marketed fertilizers, including organo-mineral fertilizers. This is due to its comprehensive composition of essential macro- and micronutrients, as evidenced by the results of the present study. The data collected also provided a thorough characterization of the frass derived from various insect species and substrates, enabling adjustments to fertilizer formulations to better align with the specific nutritional requirements of plants. Additionally, the study allowed for an in-depth understanding of compositional variations in response to the concentration of specific components in different diets. Beyond its favorable chemical composition for fertilization, the organic fertilizers developed through insect-mediated bioconversion were found to contribute significantly to soil and natural resource conservation. This aligns with the principles of sustainable agriculture by reinforcing the green paradigm, reducing the carbon footprint associated with conventional fertilizer production, and ensuring both the quality and maturity of the final product. Moreover, the findings highlight the role of insect-derived organic fertilizers in promoting the long-term sustainability of agricultural soils. DECLARATIONS a. Funding (information that explains whether and by whom the research was supported) This work was funded by SICT – Secretaria de Inovação Ciência e Tecnologia do Rio Grande do Sul, which also provided Technological and Industrial Development scholarships, in the DTI 1 and DTI 3 modalities through the TechFuturo project. It was supported by FAPERGS - Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (Edital TechFuturo) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq. Finally, this work received laboratory and analytical assistance from the Postgraduate Program in Environmental Technology at the University of Santa Cruz do Sul. b. Conflicts of interest/Competing interests (include appropriate disclosures) Declaration of Competing Interest Title: Physical-Chemical Composition Analysis of insect frass from different species produced through the bioconversion of agro-industrial waste Rafael Martins da Silva; Andreas Köhler; Daniela da Costa e Silva; Diego de Prado Vargas, Ana Lúcia Köhler. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. September 1, 2025. c. Availability of data and material (data transparency) Not applicable d. Code availability (software application or custom code) Not applicable e. Ethics approval (include appropriate approvals or waivers) Not applicable f. Authors' contributions (optional: please review the submission guidelines from the journal whether statements are mandatory) Rafael Martins da Silva : Conceptualization, Investigation, Methodology, Data collection, Writing - Original draft preparation, writing – reviewing, Editing and Statistics by Software. Andreas Köhler : Conceptualization, Supervision, Writing - Reviewing and Editing, Financial support, Diego de Prado Vargas: Methodology, Writing - Reviewing, Editing and Statistics by Software. Daniela da Costa e Silva: Methodology. Ana Köhler: Methodology. g. Consent to participate (include appropriate statements) Not applicable h. Consent for publication (include appropriate statements) Not applicable References Alidadi H, Hosseinzadeh A, Najafpoor AA, Esmaili H, Zanganeh J, Takabi MD, Piranloo FG (2016) Waste recycling by vermicomposting: Maturity and quality assessment via dehydrogenase enzyme activity, lignin, water soluble carbon, nitrogen, phosphorous and other indicators. 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Setti L, Francia E, Pulvirenti A, Gigliano S, Zaccardelli M, Pane C, Ronga D (2019) Use of black soldier fly ( Hermetia illucens (L.), Diptera: Stratiomyidae) larvae processing residue in peat-based growing media. Waste Manag. , 95 , 278-288. https://doi.org/10.1016/j.wasman.2019.06.017 Sharp RG (2013) A review of the applications of chitin and its derivatives in agriculture to modify plant-microbial interactions and improve crop yields. Agronomy, 3 (4), 757-793. https://doi.org/10.3390/agronomy3040757 Song S, Ee AWL, Tan JKN, Cheong JC, Chiam Z, Arora S, Tan HTW (2021). Upcycling food waste using black soldier fly larvae: Effects of further composting on frass quality, fertilising effect and its global warming potential. J. Clean. Prod., 288, 125664. https://doi.org/10.1016/j.jclepro.2020.125664. Tanga CM, Beesigamukama D, Kassie M, Egonyu PJ, Ghemoh, CJ, Nkoba K, Ekesi S (2022) Performance of black soldier fly frass fertiliser on maize (Zea mays L.) growth, yield, nutritional quality, and economic returns. J. Insects Food Feed, 8(2), 185-196. Terfa GN (2021) Role of black soldier fly ( Hermetia illucens ) larvae frass bio-fertilizer on vegetable growth and sustainable farming in Sub-Saharan Africa. Rev. Agric. Sci, 9 , 92-102. https://doi.org/10.7831/ras.9.0_92 van de Zande EM, Wantulla M, van Loon JJ, Dicke M (2023) Soil amendment with insect frass and exuviae affects rhizosphere bacterial community, shoot growth and carbon/nitrogen ratio of a brassicaceous plant. Plant Soil, 1-18. https://doi.org/10.1007/s11104-023-06351-6 Watson C, Preißing T, Wichern F (2021) Plant nitrogen uptake from insect frass is affected by the nitrification rate as revealed by urease and nitrification inhibitors. Front. Sustain. Food Syst, 5, 721840. https://doi.org/10.3389/fsufs.2021.721840. Xiao X, Mazza L, Yu Y, Cai M, Zheng L, Tomberlin JK, Zhang J (2018) Efficient co-conversion process of chicken manure into protein feed and organic fertilizer by Hermetia illucens L.(Diptera: Stratiomyidae) larvae and functional bacteria. J Environ Manage, 217 , 668-676. https://doi.org/10.1016/j.jenvman.2018.03.122 Yang SS, Kang JH, Xie TR, He L, Xing DF, Ren NQ, Wu WM (2019) Generation of high-efficient biochar for dye adsorption using frass of yellow mealworms (larvae of Tenebrio molitor Linnaeus) fed with wheat straw for insect biomass production. J. Clean. Prod . , 227 , 33-47. https://doi.org/10.1016/j.jclepro.2019.04.005 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-7509675\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":573162358,\"identity\":\"72b28ef9-01d0-4a9e-938a-c93abc32c6dc\",\"order_by\":0,\"name\":\"Rafael Martins da Silva\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"University of Santa Cruz do Sul\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Rafael\",\"middleName\":\"Martins da\",\"lastName\":\"Silva\",\"suffix\":\"\"},{\"id\":573162360,\"identity\":\"c0043244-5c0d-4b67-b25f-2e912521e3cc\",\"order_by\":1,\"name\":\"Andreas Köhler\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAt0lEQVRIiWNgGAWjYHACAxAhx8DA2ACkDxCvxZh0LYkNEA4RWvhnN2/78KGmLn3D7ebmDz8Y7uQT1CJx51jxzBnHDuduuHOwTbKH4ZllA0E9N3KMmXnYDuRuuJHYxszAcNiAoA55kJY//+rSDW4kNn8mSosBSAtjG3MCUEuDNFFaDG+kFTP29h02nAl0mGSPwTPCWuRuJG9m+PGtTp7vRvrjDz8q7hDWgu5OUjWMglEwCkbBKMAKAEzbQON2kibzAAAAAElFTkSuQmCC\",\"orcid\":\"\",\"institution\":\"University of Santa Cruz do Sul\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Andreas\",\"middleName\":\"\",\"lastName\":\"Köhler\",\"suffix\":\"\"},{\"id\":573162371,\"identity\":\"a46a5f9a-a13f-4058-bf59-3b6f42db31a1\",\"order_by\":2,\"name\":\"Daniela Costa\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"University of Santa Cruz do Sul\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Daniela\",\"middleName\":\"\",\"lastName\":\"Costa\",\"suffix\":\"\"},{\"id\":573162377,\"identity\":\"d6e91fcd-31e4-4bdb-980b-fab08518eda2\",\"order_by\":3,\"name\":\"Diego Prado de Vargas\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"University of Santa Cruz do Sul\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Diego\",\"middleName\":\"Prado\",\"lastName\":\"de Vargas\",\"suffix\":\"\"},{\"id\":573162392,\"identity\":\"a2a29d03-f733-48b8-95ad-70dbc76f413a\",\"order_by\":4,\"name\":\"Ana Lúcia Köhler\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Sul-MIP Biological Agents – Industry and Commerce of Biological Agents LTDA\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Ana\",\"middleName\":\"Lúcia\",\"lastName\":\"Köhler\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2025-09-01 14:53:18\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-7509675/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-7509675/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":104400939,\"identity\":\"3de78b0d-7831-46d5-ac2f-1d65d47fe3b0\",\"added_by\":\"auto\",\"created_at\":\"2026-03-11 12:11:30\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":1313657,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7509675/v1/dfcedc73-cf76-43ea-aea6-e23559f0b902.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Physical-Chemical Composition Analysis of insect frass from different species produced through the bioconversion of agro-industrial waste\",\"fulltext\":[{\"header\":\"1 Introduction \",\"content\":\"\\u003cp\\u003eCurrent projections indicate that by the year 2050, global waste production is anticipated to reach approximately 3.4 billion tons, with a predicted 44% of this being biodegradable organic matter. It is estimated that the majority of this waste will be generated in underdeveloped or developing countries (Lopes et al., 2022). In addition to the waste of organic material, which could be reused in various applications and technologies, the final disposal of this waste is carried out in landfills and open dumps (approximately 70%), having a serious impact on the environment due to the release of greenhouse gases, leachate with toxic compounds and the formation of environments conducive to vectors of pathogenic organisms (Koda et al. 2017; Lopes et al. 2022).\\u003c/p\\u003e\\n\\u003cp\\u003eThe utilization of edible insects in bioconversion represents a promising environmental technological method. This approach has the potential to reduce the volume of biodegradable organic waste, thereby eliminating the need for final disposal in landfills. Additionally, it can mitigate greenhouse gas emissions and environmental contamination by toxic leachate (Gold et al. 2018; Chiam et al. 2021). Furthermore, the bioconversion of insects such as the Black Soldier Fly (Hermetia illucens [Diptera: Stratiomyidae]), \\u003cem\\u003eTenebrio molitor\\u003c/em\\u003e (Coleoptera: Tenebrionidae), and \\u003cem\\u003eZophobas morio\\u003c/em\\u003e (Coleoptera: Tenebrionidae) has been demonstrated to be a viable solution. \\u003c/p\\u003e\\n\\u003cp\\u003eThe larvae of BSF have been shown to reduce approximately 45% to 80% of the organic waste available for bioconversion within a relatively short time frame of 2 to 5 weeks (Chiam et al. 2021; Terfa 2022; Jenkins et al. 2023). Furthermore, the employment of BSF in bioconversion or waste reduction processes exhibits enhanced efficiency in comparison to vermicomposting processes executed by worm species such as \\u003cem\\u003eEisenia foetida\\u003c/em\\u003e. This superiority can be attributed to the capacity of BSF to consume a more diverse range of waste, obviating the necessity for desalination processes of organic materials, thereby reducing operational costs and enhancing the overall practicality and viability of the process (Xiao et al. 2018). Finally, a notable benefit of employing BSF in the context of organic matter is its capacity to impede the proliferation of other fly species, which can act as vectors for pathogenic organisms (Chiam et al. 2021).\\u003c/p\\u003e\\n\\u003cp\\u003eGiven the propensity of the species \\u003cem\\u003eH. illucens\\u003c/em\\u003e, \\u003cem\\u003eT. molitor\\u003c/em\\u003e, and \\u003cem\\u003eZ. morio\\u003c/em\\u003e to reproduce more efficiently in tropical and subtropical regions, the implementation of bioconversion technology employing these insects presents a significant opportunity for innovation, particularly in developing countries (Barragan-Fonseca et al. 2017; Fuhrmann et al. 2022).\\u003c/p\\u003e\\n\\u003cp\\u003eAnother salient aspect of the bioconversion of agro-industrial waste pertains to the frass that is produced in the process (residual substrate from bioconversion or organic fertilizer). This frass is promoted by insects of various species, including \\u003cem\\u003eT. molitor\\u003c/em\\u003e, \\u003cem\\u003eZ. morio\\u003c/em\\u003e and \\u003cem\\u003eH. illucens\\u003c/em\\u003e, which have the capacity to provide nutrients necessary for the development of various crops, both organically and conventionally (Setti et al. 2019; Terfa 2021).\\u003c/p\\u003e\\n\\u003cp\\u003eResearch has been conducted to obtain simple or compound organic substrates and fertilizers from agro-industrial waste. The objective of this research is to replace synthetic and inorganic fertilizers, which have a high impact on the environment and human health, from the manufacturing process to their use. (Ronga et al. 2016; Setti et al. 2019; Poveda et al. 2019; Quilliam et al. 2020; Blakstad et al. 2023). It is crucial to acknowledge that agricultural practices reliant upon chemical fertilizers neglect the imperative of sustaining optimal levels of organic matter within the soil. This oversight directly impinges upon the preservation of the soil\\u0026apos;s physical and chemical properties, thereby exerting a detrimental influence on its capacity for productivity. Moreover, the extensive utilization of mineral fertilizers, in conjunction with their management, poses a significant threat to the sustainability of agriculture. This is due to the fact that they contribute to the progressive deterioration of soils, which can result in depletion, salinization, or desertification (Przemieniecki et al. 2021; Fuertes-Mendiz\\u0026aacute;bal et al. 2023).\\u003c/p\\u003e\\n\\u003cp\\u003eAmong the plant macronutrients, nitrogen stands out as a particularly demanding nutrient, and one that poses significant challenges in terms of supplementation. This is due in large part to the significant loss of nitrogen that occurs during the production and application of urea-based fertilizers. The loss of nitrogen in this process can range from 40% to 70%, which has far-reaching environmental consequences. The contamination of groundwater with leached nitrates is a primary concern, as it directly impacts water quality and availability. Additionally, the atmosphere becomes exposed to the release of greenhouse gases, such as NH\\u0026shy;\\u003csub\\u003e3\\u003c/sub\\u003e volatilization and N\\u003csub\\u003e2\\u003c/sub\\u003eO, which contribute to both local and global climate change. The production of ammonia-based fertilizers is a significant contributor to environmental degradation, with the production of 1,000 kg of NH\\u003csub\\u003e3\\u003c/sub\\u003e resulting in the generation of 2,000 kg of CO\\u003csub\\u003e2\\u003c/sub\\u003eeq. The process is highly dependent on natural gas, as outlined in the works of Li et al. (2021), Barbi et al. (2022), Jenkins et al. (2023).\\u003c/p\\u003e\\n\\u003cp\\u003eThe utilization of insect frass in agriculture as an organic fertilizer and phytofortifier has garnered significant attention and interest in recent years. This is due to its nutrient content and compounds that possess the potential to substitute for chemical fertilizers. Additionally, its slow-release mechanism, attributable to its low solubility in water, ensures sustained nutrient availability over extended periods (Blackstad et al. 2023). Entomology has long acknowledged the potential of insect excreta as a source of bioavailable nutrients that are readily assimilated by plants (Poveda et al. 2019).\\u003c/p\\u003e\\n\\u003cp\\u003eA limited number of studies have examined the utilization or efficacy of the organic compost produced in the bioconversion process as an organic fertilizer or soil conditioner. The findings from these studies are often inconsistent (Chiam et al. 2021). It is also important to note that the components and parameters of the frass are substrate-dependent, which complicates the comparison of results across studies due to variations in substrate source and type (Poveda et al. 2019). \\u003c/p\\u003e\\n\\u003cp\\u003eThe edible insect species \\u003cem\\u003eT. molitor\\u003c/em\\u003e produces frass, which has the potential to provide valuable nutrients for plant growth and development when used as a fertilizer or soil conditioner. The NPK balance of frass is typically 3.5-1.5-1.5 (Liu et al. 2003; Poveda et al. 2019). However, it is crucial to acknowledge that both the NPK values and the composition of other elements are contingent on the substrate (organic matter) provided during the larval development of \\u003cem\\u003eT. molitor\\u003c/em\\u003e.\\u003c/p\\u003e\\n\\u003cp\\u003eRecent studies on agricultural waste have demonstrated that this insect species possesses the capacity to survive and biodegrade organic matter, including lignin, thereby generating high-value-added products, such as larval biomass and frass. However, millions of tons of excrement waste generated are not treated properly by \\u003cem\\u003eT. molitor\\u003c/em\\u003e breeding companies, reinforcing that their correct use could significantly contribute to increasing rural incomes by using this frass as an easily available organic fertilizer (He et al. 2021). A notable advantage of \\u003cem\\u003eT. molitor\\u003c/em\\u003e and \\u003cem\\u003eZ. morio\\u003c/em\\u003e over other insect larvae, such as BSF used for food waste bioconversion processes, is that \\u003cem\\u003eT. molitor\\u003c/em\\u003e adults or \\u003cem\\u003eZ. morio\\u003c/em\\u003e beetles have fused wings/elbows and do not fly, making their biocontainment significantly easier and reducing the risk of their escape causing problems for the local ecosystem (Gan et al. 2021).\\u003c/p\\u003e\\n\\u003cp\\u003eFrass from the \\u003cem\\u003eT. molitor\\u003c/em\\u003e rearing process is a mixture of larval feces, undigested organic waste and discarded exoskeletons (Fuertes-Mendiz\\u0026aacute;bal et al. 2023). It has been demonstrated that this by-product has the capacity to increase the tolerance of different plants to abiotic stress parameters and resistance to biotic stress from different sources (Gebremikael et al. 2022). This behaviour is linked to the direct contribution of nutrients, organic carbon and, among other parameters, the presence of bioavailable chitin molecules that stimulate the immune system, as well as plant growth-promoting microorganisms (Houben et al. 2020; Barr\\u0026aacute;gan-Fonseca et al. 2022; Blakstad et al. 2023; H\\u0026eacute;nault-Ethier et al. 2023). It is important to note that the chitin in the exoskeleton of \\u003cem\\u003eT. molitor\\u003c/em\\u003e needs to be degraded by soil organisms in order to become bioavailable to the plant.\\u003c/p\\u003e\\n\\u003cp\\u003eThe conventional composting process, which can take 8 to 24 weeks, is contrasted with the accelerated composting process promoted by BSF, which takes only 5 weeks to convert organic waste into stable organic fertilizer (Anyega et al. 2021). The benefits of organic fertilizer include cost savings, environmental friendliness, and the indirect promotion of sustainable agriculture by reducing waste of organic matter (Janah et al. 2023). Furthermore, the Frass produced through bioconversion promoted by BSF accounts for 30% to 50% of the initial weight of the food substrate supplied, with this amount depending on the type and quality of the waste (Amrul et al. 2022).\\u003c/p\\u003e\\n\\u003cp\\u003eFinally, it is imperative to acknowledge the pivotal role of frass in stimulating the growth of microorganisms present in the soil, accelerating the rate of decomposition, and promoting soil respiration, nitrogen immobilization, and mineralization (van Zande et al. 2023). Moreover, extant studies have demonstrated that frass enhances plant growth, quantity, nutrient absorption, N use efficiency, and disease suppression in various crops (Quilliam et al. 2020; Anyega et al. 2021). It is noteworthy that organic fertilizers also contain nitrifying and denitrifying bacteria, which play a crucial role in the nitrogen cycle, thereby facilitating plant nitrogen uptake (Terfa 2021).\\u003c/p\\u003e\\n\\u003cp\\u003eWhile some farmers have reported the beneficial effects of excrement on plants, there are gaps in the information on the fertilizing capacity of excrement produced by insects to improve or condition soil fertility and, ultimately, plant growth. Furthermore, as emphasized by most researchers, the fertilizing potential of frass requires more research and data before the industry can expand. This is also relevant due to the search for economic and ecological alternatives to conventional mineral fertilizers, whose production depends on fossil fuels and finite resources (Poveda et al. 2019; Houben et al. 2020; He et al. 2021).\\u003c/p\\u003e\\n\\u003cp\\u003eConsequently, this study was undertaken to produce organic fertilizers from the bioconversion of agro-industrial waste promoted by different species of insects, for subsequent analysis of their quantity and quality.\\u003c/p\\u003e\"},{\"header\":\"2 Materials and Methods\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003e2.1 Rearing \\u003cem\\u003eHermetia illucens\\u003c/em\\u003e (Black Soldier Fly - BSF) larvae and obtaining frass.\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe insects were reared in the Entomology Laboratory at the University of Santa Cruz do Sul (UNISC) in air-conditioned rooms with a temperature of 26 \\u0026plusmn; 2\\u0026deg;C, relative humidity of 60 \\u0026plusmn; 5%, and a 12-hour photoperiod (adult stage). The larval stage was maintained in air-conditioned rooms with temperatures of 28\\u0026deg;C \\u0026plusmn; 2\\u0026deg;C and relative humidity of 60%. As these insects are photosensitive, they were kept in the absence of a photoperiod (dark).\\u003c/p\\u003e\\n\\u003cp\\u003eThe adults were kept in breeding cages made from white crystal organza fabric (90 cm high, 40 cm deep, and 30 cm wide) for continuous oviposition. These cages were maintained with a layer of attractant (fermented starter chick feed) and grouped strips of wood (eucalyptus, 5 cm x 20 cm x 2 cm), which served as oviposition sites.Subsequent to the eggs being laid (an average of 2 days), the wooden strips with the eggs were extracted and placed in trays with feed for the initial larvae to hatch. It is noteworthy that the oviposition wood was replenished every two days in all the rearing cages.\\u003c/p\\u003e\\n\\u003cp\\u003eDuring the growth phase, the larvae were placed in containers with different substrates, including residual whole wheat flour (E. Kuehniella rearing), orange waste, apple and banana waste (fruit mix), in different proportions (Table 1), and fed ad libitum. In the final stage (pre-pupa stage), the last-stage larvae were separated from the substrate using sieving processes to dislodge the larvae from the frass (organic fertilizer) for subsequent analysis and utilization.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.2 Breeding \\u003cem\\u003eZophobas Morio\\u003c/em\\u003e and \\u003cem\\u003eTenebrio Molitor\\u003c/em\\u003e and obtaining frass\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe rearing of both species was conducted at the Entomology Laboratory of the University of Santa Cruz do Sul (UNISC) in air-conditioned rooms with a temperature of 28 \\u0026plusmn; 2 \\u0026deg;C, relative humidity of 60 \\u0026plusmn; 5%, and a 12-hour photoperiod. The adults were maintained in plastic breeding boxes (46.7 cm x 32.3 cm x 17.9 cm) for the purpose of continuous oviposition. These boxes were maintained with a layer of substrate (wheat bran) and a water source (fruit and vegetables).\\u003c/p\\u003e\\n\\u003cp\\u003eSubsequent to egg-laying and the emergence of the first larvae, the boxes were sieved to separate the adults, eggs, and first-stage larvae. During the growth phase, the larvae of both species were placed in containers with substrate (wheat bran) and a water source, and provided with an unlimited supply of food.\\u003c/p\\u003e\\n\\u003cp\\u003eThe abiotic parameters utilized in the maintenance of the adults were employed in this instance as well. In the final stage, the last instar larvae were subjected to a sieving process to separate them from the frass (organic fertilizer), which was subsequently analyzed for its composition and quality.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 1 - Rearing parameters for the different species of bioconverter insects\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003ctable border=\\\"0\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\" width=\\\"620\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eEsp\\u0026eacute;cie\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eSubstrato\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eTemperatura\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eFotoper\\u0026iacute;odo\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eZ. morio (ZM-F)\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eFarelo de Trigo, Laranja e Chuchu\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e28 \\u0026deg;C \\u0026plusmn; 2\\u0026deg;C\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e12 h\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eT. molitor (TM-F)\\u003c/em\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eFarelo de Trigo, Laranja e Chuchu\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e28 \\u0026deg;C \\u0026plusmn; 2\\u0026deg;C\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e12h\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eH. illucens\\u003c/em\\u003e (BSF-80O)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e80 % farinha trigo integral \\u003cem\\u003e-\\u0026nbsp;\\u003c/em\\u003e20 % Laranja\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e26 \\u0026deg;C \\u0026plusmn; 2\\u0026deg;C\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eEscuro\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eH. illucens\\u003c/em\\u003e (BSF-10O)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e90 % farinha trigo integral \\u003cem\\u003e-\\u0026nbsp;\\u003c/em\\u003e10 % Laranja\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e26 \\u0026deg;C \\u0026plusmn; 2\\u0026deg;C\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eEscuro\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eH. illucens\\u003c/em\\u003e (BSF-90FM)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e10 % farinha trigo integral \\u003cem\\u003e-\\u0026nbsp;\\u003c/em\\u003e90 % Mix de Frutas\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e26 \\u0026deg;C \\u0026plusmn; 2\\u0026deg;C\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eEscuro\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e\\u003cem\\u003eH. illucens\\u003c/em\\u003e (BSF-10FM)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e90 % farinha trigo integral \\u003cem\\u003e-\\u0026nbsp;\\u003c/em\\u003e10 % Mix de Frutas\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003e26 \\u0026deg;C \\u0026plusmn; 2\\u0026deg;C\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd\\u003e\\n \\u003cp\\u003eEscuro\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.3 Compositional analysis\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.3.1 Preparation\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe sample was prepared by manually homogenizing the entire sample and reducing it by quartering until a quantity of approximately 300 g was obtained.A portion of 30 to 40 g was set aside for determining the pH of the sample in natura, and this amount was then divided into two equal fractions. One portion was allocated for granulometric analysis, employing the method delineated in the Manual of Official Analytical Methods for Fertilizers and Correctives of the Ministry of Agriculture and Livestock. The other portion was designated for chemical analysis. Subsequently, the largest portion of the sample was weighed in natura and transferred to a soil drying oven (TE-394/5 - Tecnal) at a temperature of 65 \\u0026plusmn; 5\\u0026deg;C until it attained a constant weight.\\u003c/p\\u003e\\n\\u003cp\\u003eThe fraction intended for chemical analysis was subjected to comminution and sieved through a mesh opening of 500 \\u0026micro;m.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.3.2 Moisture, total organic carbon, pH and Cation Exchange Capacity analyses\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe quantification of organic carbon was executed through the implementation of the potassium dichromate volumetric method, which entailed the wet oxidation of the organic carbon present within the sample with excess potassium dichromate and concentrated sulfuric acid, accompanied by external heating.The moisture content was determined employing the 65\\u0026deg;C moisture determination method (U65). This method involved the calculation of the percentage of moisture in the sample at 65\\u0026deg;C, utilizing the data (G1 and G2), according to the following expression: \\u003cimg src=\\\"data:image/png;base64,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\\\"\\u003ewhere: G1 = mass of the \\u0026quot;in natura\\u0026quot; sample, and G2 = mass of the sample dried at 65\\u0026deg;C (m/m).The pH was subsequently measured using the pH determination technique, which consists of suspending the sample in a 0.01 mol L\\u003csup\\u003e-1\\u003c/sup\\u003e CaCl\\u003csub\\u003e2\\u003c/sub\\u003e solution and measuring the pH potentiometrically.\\u003c/p\\u003e\\n\\u003cp\\u003eThe total nitrogen was determined by means of the Raney alloy macro-method, which consists of the ammonification of all non-ammoniacal forms of nitrogen, including organic forms. This was followed by the alkaline distillation of ammonia, which was treated with an excess amount of boric acid. The measurement of total phosphorus involved the dissolution of the phosphorus in the sample through strong acid extraction, followed by the precipitation of the orthophosphate ion as quinoline phosphomolybdate. This was then filtered, dried, and weighed. Water-soluble potassium was determined by hot extraction of water-soluble potassium and quantification by volumetry using flame photometry.\\u003c/p\\u003e\\n\\u003cp\\u003eSilicon was quantified using the ammonium molybdate spectrophotometric method after extraction with cold hydrochloric and hydrofluoric acids. The CTC/C ratio was determined by calculating the ratio between the values found for cation exchange capacity (CTC), in mmolc kg-1, and organic carbon, in mass percentage, both referred to the sample on a dry basis. The unit of measure for the CTC/C ratio is 𝑚𝑚𝑜𝑙𝑐.10\\u003csup\\u003e-1\\u003c/sup\\u003e g 𝑑𝑒 C. It is imperative to acknowledge that this ratio serves as a metric for evaluating the degree of maturity and quality of organic fertilizers.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003e2.3.1.1 Micronutrient analysis\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eAn analysis was conducted on the concentrations of phosphorus (P), potassium (K), sodium (Na), copper (Cu), calcium (Ca), iron (Fe), magnesium (Mg), zinc (Zn), and manganese (Mn). The atomic absorption spectrometry method was employed in accordance with the Manual of Official Analytical Methods for Fertilizers and Correctives of the Brazilian Ministry of Agriculture, Livestock and Supply.\\u003c/p\\u003e\"},{\"header\":\"3 Results \",\"content\":\"\\u003cp\\u003eThe results demonstrate that the total nitrogen value (N total) varies statistically according to the diet administered, the concentration of waste, and the species of insect responsible for bioconversion, as well as the statistical difference between the insects and the control (\\u003cem\\u003eEphestia kuehniella\\u003c/em\\u003e residual meal). The BSF diets with the lowest residue concentrations, BSF-10FM and BSF-10O, exhibited the highest nitrogen concentrations (4.93% and 5.07%, respectively), with no statistical difference observed. Additionally, diets with a higher amount of waste for BSF, \\u003cem\\u003eT. molitor\\u003c/em\\u003e, \\u003cem\\u003eZ. morio\\u003c/em\\u003e, and the control demonstrated no statistical difference, with an approximate result of 3.20%. It is noteworthy that all diets met the minimum levels stipulated by the Ministry of Agriculture and Livestock (MAPA) for organic fertilizers, with the frass content aligning with the established criteria.\\u003c/p\\u003e\\n\\u003cp\\u003eWith regard to phosphorus, the control showed the highest result (\\u0026cong;\\u0026nbsp;6%), followed by BSF-10FM and BSF-10O (\\u0026cong;\\u0026nbsp;5.4% and\\u0026nbsp;\\u0026cong;\\u0026nbsp;4.91%), which were statistically equal. The remaining formulations and species exhibited statistically equivalent lower results and divergent higher values, with the exception of BSF-90FM, which demonstrated the lowest value (\\u0026cong;\\u0026nbsp;2.58%) and was statistically divergent from all species and formulations.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;Potassium values were found to be comparatively low in relation to nitrogen (N) and phosphorus (P), with the exception of BSF-90FM, which exhibited the highest result (approximately 3.72%), statistically different from the others. This was followed by the control and BSF-10FM (approximately 2.43% and 2.77%, respectively). The remaining species and formulations demonstrated statistically equal values.\\u003c/p\\u003e\\n\\u003cp\\u003eMicronutrients, including calcium, magnesium, molybdenum, silicon, iron, and sulfur, exhibited comparable concentrations across all species and formulations, with minimal numerical variations. However, micronutrients such as boron, copper, cobalt, nickel, and zinc demonstrated statistical differences between species and formulations. With regard to boron, BSF-80O exhibited the highest levels (\\u0026cong;\\u0026nbsp;615.05 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e), which were statistically different from the others, followed by the formulations with 10% residue BSF-10FM, BSF-10O, and control (average of\\u0026nbsp;\\u0026cong;\\u0026nbsp;455 mg.kg-\\u003csup\\u003e1\\u003c/sup\\u003e). Among the micronutrients that demonstrated statistical differences was copper, where the highest values were obtained in the control and BSF-10FM (average of\\u0026nbsp;\\u0026cong;\\u0026nbsp;8.5 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e). Similarly, manganese exhibited higher levels in the TM-F and ZM-F species of tenebrios (417.56 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e and 318.99 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e).\\u003c/p\\u003e\\n\\u003cp\\u003eThe pH exhibited statistical differences only in BSF-80O and the control (average of\\u0026nbsp;\\u0026cong;\\u0026nbsp;7.3) in comparison to the other species and formulations. Total carbon levels in all the formulations demonstrated levels that exceeded the minimum requirement (15% MAPA reference), with the lowest levels observed in BSF-10O and the control (38.61% and 36.22%, respectively). The cation exchange capacity (CEC) exhibited higher values in the BSF-10FM and BSF-90FM samples (706.32 mmol.kg\\u003csup\\u003e-1\\u0026nbsp;\\u003c/sup\\u003eand 594.76 mmol.kg\\u003csup\\u003e-1\\u003c/sup\\u003e), respectively, both of which incorporated multiple fruits in their formulation (banana and apple).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 1 - Composition of the frass produced by each species (MAPA)\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003ctable border=\\\"0\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\" width=\\\"558\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eParameters\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eMin. Levels\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eTM-F\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eZM-F\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF-10 FM\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF-90FM\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF-10O\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF-80O\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eControl\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eGranulometry\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003eClassifica\\u0026ccedil;\\u0026atilde;o\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003ePowder\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003ePowder\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003eBran\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003eBran\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003eBran\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003eBran\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003eBran\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eMax humidity\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e40.00%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e6.79a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e9.51b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e7.67a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e7.07a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e13.69d\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n 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\\u003cp\\u003e0.020%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e12.93a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e15.35b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e18.5c\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e15.05d\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e14.64e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e13.2a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e18.72\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eFe (%)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e0.020%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e0.02a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e0.03a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e0.03a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e0.03a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e0.03a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e0.02a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e0.03a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eMn (mg/Kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e0.020%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e314.73a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e346.18b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e273.62c\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e121.84d\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e120.91d\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e51.28e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e255.26c\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eMo (mg/Kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e0.005%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;15a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;15a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;15a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;15a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;15a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;15a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;15a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eNi (mg/Kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e0.005%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e2.4b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e3c\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e3c\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e2.2b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e2.5b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eSe (mg/Kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e0.003%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eSi (%)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e0.050%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e0.03a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e0.06a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e0.06a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e0.06a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e0.03a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e0.03a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e0.02a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 95px;\\\"\\u003e\\n \\u003cp\\u003eZn (mg/Kg)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 78px;\\\"\\u003e\\n \\u003cp\\u003e0.100%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 53px;\\\"\\u003e\\n \\u003cp\\u003e110.42a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 54px;\\\"\\u003e\\n \\u003cp\\u003e118.06a\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e129.81b\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e68.06c\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e100e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 57px;\\\"\\u003e\\n \\u003cp\\u003e47.74d\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"top\\\" style=\\\"width: 52px;\\\"\\u003e\\n \\u003cp\\u003e122.43ab\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\\n\\u003cp\\u003eTo comprehensively assess the various organic fertilizers derived from bioconversion, it was imperative to analyze the contaminants, as mandated by MAPA, Brazil\\u0026apos;s regulatory authority. As delineated in Table 2, the frass from all species and dietary regimens exhibited negligible levels of contamination by heavy metals, including arsenic, cadmium, lead, hexavalent chromium, and mercury. With regard to nickel, it was detected above the minimum analysis limits in the organic fertilizers, with the exception of the TM-F frass (\\u0026lt;2 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e), but all were well below the contamination levels (70 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;It is imperative to note that the maximum tolerance level for thermotolerant coliforms is 1000 NMP.g\\u003csup\\u003e-1\\u003c/sup\\u003e. However, it is noteworthy that with the exception of the TM-F sample (450 NMP.g\\u003csup\\u003e-1\\u003c/sup\\u003e), all the samples were found to be free of these. Furthermore, the tests revealed the absence of helminth eggs and Salmonella sp. in all the frass samples across all species and diets. Finally, it is noteworthy that all the organic fertilizer samples were found to be free from contamination, thereby meeting the conditions for use as stipulated by Brazilian legislation in accordance with MAPA standards and legislation from other countries.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 2 - Limits of contaminants allowed by Brazilian legislation (MAPA)\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003ctable border=\\\"0\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\" width=\\\"576\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eContaminants\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eAllowed Parameters\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eTM-F\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eZM-F\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF-80O\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF-10O\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF-90 FM\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF-10 FM\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eAs\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e20 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eCd\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e3 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003ePb\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e150 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eCr-VI\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e2 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.4\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.4\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.4\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.4\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.4\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.4\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eHg\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e1 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;0.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eNi\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e70 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;2.0\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e2.4\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e2.5\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e2.2\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e3\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e3\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eSe\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e80 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;1\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eHelmints Eggs\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e1/4g\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003eNegative\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003eNegative\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003eNegative\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003eNegative\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003eNegative\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003eNegative\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eTtC\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003e1000\\u0026nbsp;\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e450\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;180\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;180\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;180\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003e140\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003e\\u0026lt;180\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 96px;\\\"\\u003e\\n \\u003cp\\u003eSalmonella sp.\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 93px;\\\"\\u003e\\n \\u003cp\\u003eAbsence\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003eAbsence\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003eAbsence\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003eAbsence\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 63px;\\\"\\u003e\\n \\u003cp\\u003eAbsence\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 70px;\\\"\\u003e\\n \\u003cp\\u003eAbsence\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd valign=\\\"bottom\\\" style=\\\"width: 62px;\\\"\\u003e\\n \\u003cp\\u003eAbsence\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n\\u003c/table\\u003e\"},{\"header\":\"4 Discussion\",\"content\":\"\\u003cp\\u003eThe waste degradation efficiency and bioconversion periods of insect species vary, as does the amount of frass fertilizer available. In general, the larvae of \\u003cem\\u003eH. illucens\\u003c/em\\u003e exhibit a high waste degradation efficiency (55-80%) (Lalander et al. 2015; Beesigaukama et al. 2022; da Costa e Silva et al. 2024), a relatively brief bioconversion time, and consequently, they produce higher quantities of organic fertilizer in comparison to other insect species. Conversely, \\u003cem\\u003eT. molitor\\u003c/em\\u003e and \\u003cem\\u003eZ. morio\\u003c/em\\u003e require 4 to 6 weeks to convert organic waste into fertilizer, yielding comparatively lower quantities of frass. Lignocellulosic materials impose a significant stressor on the digestive system of the BSF, resulting in a diminished concentration of organic matter in frass (Manurung et al. 2016). The degradation and conversion of lignocelluloses by BSF is considered viable but inefficient (Liu et al. 2021a).\\u003c/p\\u003e\\n\\u003cp\\u003eThe characteristics of insect frass, including its composition and quality, exhibit variability due to the substrate provided to the larvae during the rearing process and subsequent treatment. This variability may be attributed to legislative and/or health-related recommendations (Klammsteiner et al. 2020; Lomonaco et al. 2024). The average concentration of carbon (C) has been documented to range from 35% to 40% (Chiam et al. 2021; Watson et al. 2021; Song et al. 2021; Lopes et al. 2022), which is in accordance with the findings of this study, which identified concentrations in all species and different diets that varied between 38% and 48%, inferring that the larvae have the ability to concentrate organic matter and accumulate nutrients, even if the frass is the product of their excretion plus the remains of the substrate they are fed, since the control had lower amounts of C (\\u0026cong;\\u0026nbsp;36%).\\u003c/p\\u003e\\n\\u003cp\\u003eThe concentrations of nitrogen (N), phosphorus (P), and potassium (K) found in these species are comparable to those found in farmyard manure, particularly poultry manure (Poveda et al. 2019), thereby substantiating their considerable fertilizer potential. In contrast to conventional mineral fertilizer, frass also contains trace amounts of micronutrients (e.g., copper [Cu], zinc [Zn], manganese [Mn], and iron) that can be particularly advantageous for plant growth. The carbon (C) fraction in frass is comparable to that of pig and poultry manure (Roth\\u0026eacute; et al. 2019; Liu et al. 2019), with a substantial soluble C fraction, which may be associated with the efficient digestion of cellulose and lignin compounds by \\u003cem\\u003eT. molitor\\u003c/em\\u003e, \\u003cem\\u003eZ. morio\\u003c/em\\u003e, and BSF larvae (Houben et al. 2020). It is noteworthy that all types of frass from diverse species and substrates exhibited the majority of essential trace elements for plant growth, with the exception of Ni.\\u003c/p\\u003e\\n\\u003cp\\u003eThe nitrogen (N) concentrations did not exhibit significant variation across species or dietary treatments (approximately 3%), suggesting that the diets did not substantially influence the nitrogen content in frass. These findings align with those reported by Menino et al. (2021), Watson et al. (2021), Anyega et al. (2021), Green (2023), and Lomonaco et al. (2024) (Table 3). However, in the present study, higher N concentrations were observed in BSF diets supplemented with only 10% waste, which contained elevated levels of starch and carbohydrates due to the inclusion of flour. Previous studies have suggested that the incorporation of protein into diets can enhance nitrogen concentrations in frass (Setti et al. 2019). Nevertheless, it is well established that larvae generally exhibit suboptimal development on diets rich in protein but deficient in carbohydrates, which in turn leads to insufficient frass production due to nutrient limitations (da Costa e Silva et al. 2024). Furthermore, Sarpong et al. (2019) reported that diets characterized by high protein and sugar/starch content, coupled with low fiber concentrations, promote the accumulation of nutrients in frass.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTable 3 - Results of recent research on biofertilizers produced or BSF bioconversion.\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cdiv align=\\\"Left\\\"\\u003e\\n \\u003ctable border=\\\"1\\\" cellspacing=\\\"0\\\" cellpadding=\\\"0\\\" width=\\\"559\\\"\\u003e\\n \\u003ctbody\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\u003cbr\\u003e\\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e(Chiam et al., 2021) BSF - Fresh Okara\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e(Menino et al., 2021)\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF - Potato and Onion\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e(Watson et al., 2021)\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e\\u003cem\\u003eT.molitor\\u003c/em\\u003e\\u003c/strong\\u003e\\u003cstrong\\u003e\\u0026nbsp;- Insectus Mealworm Grow\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e(G\\u0026auml;rttling et al., 2020) BSF - Maize\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e(Song et al., 2021)\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003eBSF \\u0026ndash; Okara and wheat bran\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e(Anyega et al., 2021) BSF \\u0026ndash; Brewer\\u0026acute;s spent grains\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e\\u003cstrong\\u003e(Kawasaki et al., 2020) BSF \\u0026ndash; Household organic waste\\u003c/strong\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003eC\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e37,07%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e38,7%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e35,7%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e41,8%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e37,1%\\u003c/p\\u003e\\n 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mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003eFe\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e3,69%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e896 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e15 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n 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\\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e2,77 \\u0026nbsp;mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003eZn\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e1,73%)\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e137 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e15 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e0,10 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e182 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e10 mg.kg\\u003csup\\u003e-1\\u003c/sup\\u003e\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003eNi\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e0,11%\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003eAs\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003eCd\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003ePb\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003eCr\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e-\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003ctr\\u003e\\n \\u003ctd style=\\\"width: 30px;\\\"\\u003e\\n \\u003cp\\u003eC/N\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\u003cbr\\u003e\\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e13,8\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 85px;\\\"\\u003e\\n \\u003cp\\u003e16\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e12,6\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 66px;\\\"\\u003e\\n \\u003cp\\u003e7,76\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e10,7\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003ctd style=\\\"width: 76px;\\\"\\u003e\\n \\u003cp\\u003e16,6\\u003c/p\\u003e\\n \\u003c/td\\u003e\\n \\u003c/tr\\u003e\\n \\u003c/tbody\\u003e\\n \\u003c/table\\u003e\\n\\u003c/div\\u003e\\n\\u003cp\\u003e\\u0026nbsp;In general, the predominant form of nitrogen in black soldier fly (BSF) frass is ammonium nitrogen (NH₄⁺-N), while nitrate nitrogen (NO₃⁻-N) concentrations tend to be lower. This is likely due to the fact that BSF frass is processed solely through the larval digestive system, in contrast to fertilizers derived from animals such as poultry and cattle, where the substrate undergoes microbial fermentation during digestion. Moreover, insect frass, unlike conventional livestock manure, represents a novel fertilizer with a lower potential for NO₃⁻-N accumulation in plants, thereby reducing the risk of nitrate-induced toxicity (Kawasaki et al., 2020).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;Beyond nitrogen content, the nutritional advantages of insect frass extend to other essential elements, as evidenced by increased uptake of phosphorus (P), potassium (K), magnesium (Mg), and sulfur (S) in plants fertilized with frass compared to those treated with conventional organic fertilizers. Additionally, the elevated phosphorus concentrations in frass may enhance nitrogen assimilation in plants, as phosphorus plays a crucial role in energy transfer processes (Fageria 2001; Kawasaki et al. 2020).\\u003c/p\\u003e\\n\\u003cp\\u003eAlthough the total carbon content remained consistent across all frass samples, the C/N ratio varied among them, with values of 13.20 for TM-F, 15.84 for TM-Z, 8.46 for BSF-10FM, 13.63 for BSF-90FM, 7.61 for BSF-10O, and 15.84 for BSF-80O. A C/N ratio exceeding 20 presents a risk of nitrogen immobilization in the soil; however, the nitrogen content in frass can benefit plants with efficient nitrogen utilization, which is largely influenced by their rhizobiome (Klammsteiner et al. 2020; Dzepe et al. 2022). Furthermore, insect larvae undergo six developmental instars, during which they periodically shed their exoskeleton. This exoskeleton is primarily composed of chitin, a polymer of N-acetylglucosamine (C\\u003csub\\u003e8\\u003c/sub\\u003eH\\u003csub\\u003e13\\u003c/sub\\u003eO\\u003csub\\u003e5\\u003c/sub\\u003eN)\\u003csub\\u003en\\u003c/sub\\u003e, which affects the C/N ratio. Additionally, chitin degradation can yield chitosan, a compound with plant growth-promoting properties that also enhances resistance to pathogens (Sharp 2013; Klammsteiner et al. 2020; Nayak et al. 2020; Fuertes-Mendiz\\u0026aacute;bal et al. 2023). Supporting this premise, as well as the findings of the present study, Anyega et al. (2020) suggests that the increased growth rates and yields observed in vegetables cultivated with BSF frass may be attributed to its advanced maturity and stability, as evidenced by a lower C/N ratio. The pH variation observed in the BSF frass samples was minimal (6.00\\u0026ndash;7.57), exhibiting a slight tendency toward alkalinity. In contrast, the pH values of TM-F and ZM-F remained nearly constant (5.75\\u0026ndash;5.78), indicating a slightly more acidic nature. The average pH value reported in the literature is 7.46, which is also mildly alkaline and, as observed in this study, represents one of the least variable parameters. Furthermore, it is well established that BSF, in its organic form, actively alters the pH of the substrate toward alkaline levels, thereby supporting the findings of this study (G\\u0026auml;rttling \\u0026amp; Schulz 2022). Additionally, the higher nutrient concentrations and lower C/N ratio associated with organic insect-derived fertilizers underscore the remarkable efficiency of BSF, \\u003cem\\u003eT. molitor\\u003c/em\\u003e, and \\u003cem\\u003eZ. morio\\u003c/em\\u003e larvae in nutrient recycling (Lalander et al. 2015; Tanga et al. 2022).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp; Compost or biofertilizer maturity refers to the degree of completeness of the bioconversion process, characterized by the absence of phytotoxic compounds as well as animal and plant pathogens that could negatively impact seed germination, plant growth, and soil health. In simple terms, compost maturity signifies its suitability for agricultural application as a fertilizer, assessed through chemical, physical, and biological indicators. During the present study, the C/N ratio ranged from 7 to 16, indicating high-quality and mature compost (Goyal et al. 2015; Houben et al. 2020; da Costa e Silva et al. 2024). Similarly, the pH values of all analyzed species and substrates remained within the range of 6.0 to 7.5, a threshold considered acceptable for mature compost. This finding aligns with the observations of Lomonaco et al. (2024), who reported that \\u003cem\\u003eHermetia illucens\\u003c/em\\u003e tends to shift the pH of frass toward more alkaline conditions. Furthermore, the alkalization of frass induced by BSF-driven bioconversion is attributed to the release of ammonium ions (NH₄⁺) and ammonia (Alidadi et al. 2020; Lomonaco et al. 2024). It is important to highlight that the chemical parameters, including macro- and micronutrient concentrations, align with the standards recommended for high-quality fertilizers (Table 1). These fertilizers ensure the release of adequate nutrient levels necessary for optimal plant development. Furthermore, the observed values comply with the regulatory standards established in various countries, including Brazil, the United States, Canada, African nations, and the European Union.\\u003c/p\\u003e\\n\\u003cp\\u003eThe effectiveness of organic fertilizers in vegetable production is closely associated with their nutrient composition, particularly nitrogen (N), and the rate at which these nutrients are released (Cabrera et al. 2005; Fuertes-Mendiz\\u0026aacute;bal et al. 2023). The frass derived from \\u003cem\\u003eTenebrio molitor\\u003c/em\\u003e, \\u003cem\\u003eZophobas morio\\u003c/em\\u003e, and black soldier fly (BSF) exhibits significant potential as an organic fertilizer due to its high concentrations of nitrogen (N), phosphorus (P), and potassium (K), which are comparable to those found in other organic fertilizers, such as raw manure (Houben et al. 2020; Dzepe et al. 2022). This observation aligns with the findings of the present study, as the frass of these insect species contained average N-P-K values of approximately 3% N, 4% P₂O₅, and 1.80% K₂O, respectively. Additionally, Fuertes-Mendiz\\u0026aacute;bal et al. (2023) reported a substantial increase in lettuce leaf biomass with the application of only 1% frass, attributed to enhanced nitrogen uptake. Notably, in contrast to conventional organic fertilizers, insect frass is characterized by a high content of labile organic matter measured at approximately 40% in all species and substrates examined in this study and exhibits rapid mineralization kinetics (Chavez a\\u0026amp; Uchanski 2021), thereby facilitating the swift availability of nutrients.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;Studies have demonstrated that the application of \\u003cem\\u003eHermetia illucens\\u003c/em\\u003e and \\u003cem\\u003eTenebrio molitor\\u003c/em\\u003e frass as fertilizer results in higher yields and improved nutritional quality in various crops, including maize, tomatoes, cabbage, cowpeas, pepper, shallots, and barley, when compared to conventional fertilizers (Tanga et al. 2021; Beesigamukama et al. 2022). Moreover, soil amendment with \\u003cem\\u003eH. illucens\\u003c/em\\u003e and \\u003cem\\u003eT. molitor\\u003c/em\\u003e frass has been shown to suppress the proliferation of soil-borne pathogens, enhance soil microbial activity, reduce soil acidity and salinity, improve nitrogen mineralization, and increase nutrient availability, thereby contributing to overall soil quality and promoting plant growth (Houben et al. 2020; Barrag\\u0026aacute;n-Fonseca et al. 2022).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;Additionally, fertilization with black soldier fly (BSF) frass has been observed to enhance potassium (K) uptake by plants, where potassium serves as an enzyme activator essential for maintaining cell membrane integrity and developmental potential (Hellgren et al. 2006; Radzikowska-Kujawska et al. 2023). Under optimal growth conditions, phosphorus (P) absorption is also notably increased (Radzikowska-Kujawska et al. 2023). Furthermore, ongoing research in the literature explores the role of polyphenolic compounds absorbed by plants, particularly those fertilized with insect frass. Among these compounds, ferulic acid has garnered attention due to its efficient absorption from food, its metabolic pathways in the human body, and its potential health-promoting properties (Kwee et al. 2011; Radzikowska-Kujawska et al. 2023).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp; In the European Union, the classification and labeling of frass as an organic fertilizer are regulated under Regulation (EU) 2019/1009, which generally categorizes it as a solid organic fertilizer (PFC 1 (A) (I)). Moreover, this regulation establishes that compound solid organic fertilizers must contain a minimum of 1% total nitrogen (N), phosphorus pentoxide (P₂O₅), or potassium oxide (K₂O), with the combined proportion of these primary nutrients amounting to at least 4% by mass (Houben et al. 2020; IPIFF,2021). In this context, it is important to highlight that the frass derived from all species and diets examined in the present study complies with the regulatory requirements set forth by both the Brazilian Ministry of Agriculture and Livestock and the European Union.\\u003c/p\\u003e\\n\\u003cp\\u003eFurthermore, the substitution of traditional organic waste treatment methods, such as aerobic composting, with the use of black soldier fly (BSF) larvae has demonstrated a significant reduction in the global warming potential associated with organic waste management, decreasing emissions by approximately 50% (Mertenat et al. 2019).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp; \\u0026nbsp; Despite its potential benefits, the application of insect frass as a fertilizer remains an area requiring further investigation. This necessity arises primarily due to the volatilization of nitrogen into potentially toxic forms, which is influenced by soil pH and cation exchange capacity (CEC). Additionally, regulatory frameworks, such as those of the European Union, mandate the standardization of frass through thermal treatment at 70\\u0026deg;C, which may further affect its properties and efficacy as a fertilizer. Consequently, the optimization of fertilizer application methods will be a critical consideration in future research and agricultural practices (Kawasaki et al. 2020; Houben et al. 2020; G\\u0026auml;rttling \\u0026amp; Schulz 2022).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp;\\u003c/p\\u003e\"},{\"header\":\"5 Conclusion\",\"content\":\"\\u003cp\\u003eInsect frass has demonstrated significant potential as a sustainable organic fertilizer, with the capacity to fully replace conventionally marketed fertilizers, including organo-mineral fertilizers. This is due to its comprehensive composition of essential macro- and micronutrients, as evidenced by the results of the present study. The data collected also provided a thorough characterization of the frass derived from various insect species and substrates, enabling adjustments to fertilizer formulations to better align with the specific nutritional requirements of plants. Additionally, the study allowed for an in-depth understanding of compositional variations in response to the concentration of specific components in different diets.\\u003c/p\\u003e\\n\\u003cp\\u003eBeyond its favorable chemical composition for fertilization, the organic fertilizers developed through insect-mediated bioconversion were found to contribute significantly to soil and natural resource conservation. This aligns with the principles of sustainable agriculture by reinforcing the green paradigm, reducing the carbon footprint associated with conventional fertilizer production, and ensuring both the quality and maturity of the final product. Moreover, the findings highlight the role of insect-derived organic fertilizers in promoting the long-term sustainability of agricultural soils.\\u003c/p\\u003e\"},{\"header\":\"DECLARATIONS \",\"content\":\"\\u003cp\\u003ea. Funding (information that explains whether and by whom the research was supported)\\u003c/p\\u003e\\n\\u003cp\\u003eThis work was funded by SICT – Secretaria de Inovação Ciência e Tecnologia do Rio Grande do Sul, which also provided Technological and Industrial Development scholarships, in the DTI 1 and DTI 3 modalities through the TechFuturo project. It was supported by FAPERGS - Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (Edital TechFuturo) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq. Finally, this work received laboratory and analytical assistance from the Postgraduate Program in Environmental Technology at the University of Santa Cruz do Sul.\\u003c/p\\u003e\\n\\u003cp\\u003eb. Conflicts of interest/Competing interests (include appropriate disclosures)\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eDeclaration of Competing Interest\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eTitle: Physical-Chemical Composition Analysis of insect frass from different species produced through the bioconversion of agro-industrial waste\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eRafael Martins da Silva; Andreas Köhler; Daniela da Costa e Silva; Diego de Prado Vargas, Ana Lúcia Köhler.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp; \\u0026nbsp;\\u0026nbsp;The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\\u003c/p\\u003e\\n\\u003cp\\u003eSeptember 1, 2025.\\u003c/p\\u003e\\n\\u003cp\\u003ec. Availability of data and material (data transparency)\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\\n\\u003cp\\u003ed. Code availability (software application or custom code)\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\\n\\u003cp\\u003ee. Ethics approval (include appropriate approvals or waivers)\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\\n\\u003cp\\u003ef. Authors' contributions (optional: please review the submission guidelines from the journal whether statements are mandatory)\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eRafael Martins da Silva\\u003c/strong\\u003e: Conceptualization, Investigation, Methodology, Data collection, Writing - Original draft preparation, writing – reviewing, Editing and Statistics by Software. \\u003cstrong\\u003eAndreas Köhler\\u003c/strong\\u003e: Conceptualization, Supervision, Writing - Reviewing and Editing, Financial support, \\u003cstrong\\u003eDiego de Prado Vargas:\\u003c/strong\\u003e Methodology, Writing - Reviewing, Editing and Statistics by Software. \\u003cstrong\\u003eDaniela da Costa e Silva:\\u003c/strong\\u003e Methodology. \\u003cstrong\\u003eAna Köhler:\\u003c/strong\\u003e Methodology.\\u003c/p\\u003e\\n\\u003cp\\u003eg. Consent to participate (include appropriate statements)\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\\n\\u003cp\\u003eh. Consent for publication (include appropriate statements)\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eAlidadi H, Hosseinzadeh A, Najafpoor AA, Esmaili H, Zanganeh J, Takabi MD, Piranloo FG (2016) Waste recycling by vermicomposting: Maturity and quality assessment via dehydrogenase enzyme activity, lignin, water soluble carbon, nitrogen, phosphorous and other indicators. 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Sci, \\u003cem\\u003e9\\u003c/em\\u003e, 92-102. https://doi.org/10.7831/ras.9.0_92\\u003c/li\\u003e\\n\\u003cli\\u003evan de Zande EM, Wantulla M, van Loon JJ, Dicke M (2023) Soil amendment with insect frass and exuviae affects rhizosphere bacterial community, shoot growth and carbon/nitrogen ratio of a brassicaceous plant. Plant Soil, 1-18. https://doi.org/10.1007/s11104-023-06351-6\\u003c/li\\u003e\\n\\u003cli\\u003eWatson C, Prei\\u0026szlig;ing T, Wichern F (2021) Plant nitrogen uptake from insect frass is affected by the nitrification rate as revealed by urease and nitrification inhibitors. Front. Sustain. Food Syst, 5, 721840. https://doi.org/10.3389/fsufs.2021.721840.\\u003c/li\\u003e\\n\\u003cli\\u003eXiao X, Mazza L, Yu Y, Cai M, Zheng L, Tomberlin JK, Zhang J (2018) Efficient co-conversion process of chicken manure into protein feed and organic fertilizer by \\u003cem\\u003eHermetia illucens\\u003c/em\\u003e L.(Diptera: Stratiomyidae) larvae and functional bacteria. J Environ Manage, \\u003cem\\u003e217\\u003c/em\\u003e, 668-676. https://doi.org/10.1016/j.jenvman.2018.03.122\\u003c/li\\u003e\\n\\u003cli\\u003eYang SS, Kang JH, Xie TR, He L, Xing DF, Ren NQ, Wu WM (2019) Generation of high-efficient biochar for dye adsorption using frass of yellow mealworms (larvae of \\u003cem\\u003eTenebrio molitor\\u003c/em\\u003e Linnaeus) fed with wheat straw for insect biomass production. J. Clean. Prod\\u003cem\\u003e.\\u003c/em\\u003e, \\u003cem\\u003e227\\u003c/em\\u003e, 33-47. https://doi.org/10.1016/j.jclepro.2019.04.005\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":true,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research Square\",\"twitterHandle\":\"researchsquare\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":false,\"inReviewRevisionsEnabled\":true},\"keywords\":\"Residues, Fertilizer, Entomology, Regulation\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-7509675/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-7509675/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eThe objective of the present study was to develop organic fertilizers through the bioconversion of agro-industrial waste, utilizing various insect species, with the intent of evaluating both the quantity and quality of the resulting products. To conduct the research, a controlled rearing of the insect species \\u003cem\\u003eTenebrio molitor\\u003c/em\\u003e, \\u003cem\\u003eZophobas morio\\u003c/em\\u003e, and \\u003cem\\u003eHermetia illucens\\u003c/em\\u003e was established under laboratory conditions, incorporating diverse substrates and varying concentrations to assess the composition of the frass produced. The physicochemical characterization included analyses of moisture content, total organic carbon, pH, cation exchange capacity, and micronutrient determination via atomic absorption spectrometry, in compliance with current regulations for organic fertilizers. The primary plant nutrients, such as total carbon (36-44%), nitrogen (2.7-5%), phosphorus (2.5-6%), and potassium (1.7-3.72%), exhibited variation depending on the substrate and insect species used, with macro and micronutrient values in alignment with the requirements set forth by Brazilian and European Union legislation. Furthermore, no contaminants were detected in any of the samples analyzed. It can be concluded that the frass produced by \\u003cem\\u003eT. molitor\\u003c/em\\u003e, \\u003cem\\u003eZ. morio\\u003c/em\\u003e, and \\u003cem\\u003eH. illucens\\u003c/em\\u003e demonstrates significant potential as an organic fertilizer, as its concentrations of nitrogen, phosphorus, and potassium are comparable to those found in other organic fertilizers on the market. This suggests its potential to fully replace both traditional commercial fertilizers and organomineral fertilizers, thereby contributing to a reduction in the carbon footprint associated with fertilizer production.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Physical-Chemical Composition Analysis of insect frass from different species produced through the bioconversion of agro-industrial waste\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2026-01-14 12:19:21\",\"doi\":\"10.21203/rs.3.rs-7509675/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"researchsquare\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":true,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"/submission\",\"title\":\"Research Square\",\"twitterHandle\":\"researchsquare\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":false,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"3c7a7074-0e51-4053-a8e7-82ad9a6d0a99\",\"owner\":[],\"postedDate\":\"January 14th, 2026\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2026-03-03T11:42:27+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2026-01-14 12:19:21\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-7509675\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-7509675\",\"identity\":\"rs-7509675\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}