Mealworm frass is a potential biofertilizer.

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The increase of rearing insects will be associated with generating high quantities of frass.Before a major upscaling of the industry takes place, it is necessary to find solutions that allow efficient valorization of these by-products.The purpose of the study is to investigate the potential of frass.A pot experiment was established and the soil was amended with mealworm or a mixture of 50% NPK and 50% fras.Changes of soil properties and growth were analyzed.frass is similar to mineral NPK in its ability to improve the amount of N, P and K in the soil.Watersoluble P concentration is five times lower in the presence of frass, which prevents P from loss and sorption onto soil.The addition of frass stimulates soil microbial activity, especially when it is mixed with mineralfertilizer, suggesting a synergistic effect between both amendments.Our results show that frass can be used as a partial or complete substitute for mineral NPK.This is relevant because of a reduced availability of mineral fertilizers and being consistent with the principles of the circular economy.

Reducing agriculture's negative environmental impact is one of the greatest challenges that the twenty-first century has to offer.Ensuring sustainable food production for 2.3 billion more people in the next four decades is the goal of societal transition and industrial transformation.If we want to restrict agricultural land use and avoid environmental degradation, we need to look for alternatives to conventional sources of meat, which is expected to increase from current levels by more than 75% in 2050.Eco-industrial development approach aims at closing economic and ecological loops of resource flows by reducing the use of natural resources.

Rearing insects for mass consumption is sparking interest due to their high nutrition and efficient use of resources.As recently reviewed by Dicke10, insect production has a much smaller ecological footprint in terms of land and water use and greenhouse warming potential compared to the production of chicken, pigs and cattle which is related to its much lower feed to meat conversion ratio.In addition to being an alternative source of proteins for humans and animals, insect farming has also been considered to produce biofuel, some rare lipids and chitin, which has a great economical value due to its application in food, cosmetics, pharmaceuticals, textile etc.

There have been major efforts to promote the use of insects as a source of food, spurred in large part by the Food and Agriculture Organization (FAO) of the United Nations work examining the multiple dimensions of insect rearing as an important future food source.The legal status of insects and their derived products was clarified in the European Union.Several EU members.France and Germany have partially legalized the production and commercialization of insects.The implementation of insect producers has been boosted by these incentive policies.nsect, a French insect farming startup, has recently raised $125 million in Series C funding to develop their activities, which is the largest early-stage agtech funding deal on record in Europe.The number of jobs that will be created by the development of this sector is expected to increase in the EU.

A waste stream consisting of moulting skins (exuviae) and insect faeces is produced by insect production.Due to its high labile C content, frass deposition to soil has a great impact on soil fertility.Several companies have already decided to sell frass.Even though some farmers have reported beneficial effects of frass to plants, there is currently very limited information on the ability of insect farms to improve soil fertility and plant growth.The potential of frass needs urgent research before a major upscaling of the industry takes place.Given the urgent need to find cost-effective and environmental-friendly alternatives to conventional mineral fertilizers, this would be relevant.

The potential of frass from mealworm as afertilizer is evaluated by investigating its impact on soil biological activity and crop growth.

The high concentrations of N, K and P found in frass confirm its high fertilization potential.frass contained small concentrations of micronutrients.It may be beneficial for crops.There is an image of frass.The structure of frass was revealed while the map was being created.The distribution of P, K and Ca within the frass organic matter was shown to be uniform by the C and O maps.The release of nutrients by frass after they are incorporated into the soil should be homogeneity and possibly more extended than in the case of unevenly distributed nutrients.The C fractionation in frass was similar to that of poultry manure, with high and low C fractions.The high fraction of labile C is in agreement with previous findings and may be related to the high digestion of cellulose and lignin compounds by mealworm.

Our data showed that frass from the mealworm farm was quickly deposited into the soil after its decomposition in an early process.There are more than one 3).37% of the organic carbon was mineralized after 7 days.After 91 days, a slower but continuous C was present.After 17 days of incubation, Nitrogen had a similar pattern, with a rapid mineralization just after frass application, followed by a slower but continuous N mineralization.The mealworm frass used in this study compares well with other insects.In a 120-day experiment, Lovett and Ruesink17 reported that the best gypsies were found in the first 10 days.The mealworm frass used in this study had a high labile C concentration and this likely stimulated microbial growth, resulting in high rate of C and N.The high labile C concentration of poultry manure, which had a similar chemical composition than frass, was intense in the early stages of incubation.

The measurement of average well color development (AWCD) and S and H indices were used to assess microbial metabolism.4.AWCD can be used as an indicator of soil microbial activity.The organic amendments are a source of energy.The supply of organic matter to soil through different amendments generally stimulates microbial activity and increases the microbial community functional potential, which can be reflected by an increase of AWCD and R and H indices32.Similar to other organic amendments, the inclusion of frass resulted in an increase of the AWCD and/or S indices compared to the NPK treatment.The results show that the AWCD and diversity indices were increased by the addition of organic C, alone or in combination with NPK.The low rate of frass application in the present study might explain the lack of improvement of S index.10 Mg haEven though organic amendments increased AWCD at both 10 and 20 Mg ha1 application rate, S index was only improved in the 20Mg ha'1 treatment due to higher C availability for microorganisms.S and H were the highest in the 50NPK/50 Frass treatment.According to Hu et al.34, higher diversity in soil amended with a combination of both organic and mineral fertilizers is due to a larger number of C sources under this treatment.The benefits of using frass and NPK on soil microbial functional diversity suggest that a partial substitution of mineral fertilization is an attractive option for sustainable management.Our study only focused on soil organisms.The next challenge will be to investigate the effect of frass on soil fauna as it also plays a key role on the regulation of ecosystem functioning.

The average well-color development (AWCD), Richness (S) and Shannon-Weaver index (H) of metabolized substrates are in BIOLOG EcoPlate after frass application.The values are average.At the 5% level, columns with the same letter don't differ much.

Although the soil pH of the frass treatment was not as low as the NPK treatment, it was still slightly acidic, most likely due to its rapid decomposition.5).The fast nitrification was reported in the experiment.The frass treatment may have an additional source of protons.The highest available K and P concentrations were measured in the NPK treatment, which was obviously due to the supply of these nutrients in a soluble form, even though all the fertilizer treatments added the same total amounts of P and K.5).The frass treatment had more available K and P concentrations than the control, suggesting that the treatment gave the nutrients in a readily available form.The lower K and P concentrations in the presence of frass compared to the NPK treatment is in agreement with other studies.Houben et al.22 found that after 100 days, available P concentration in a soil amended with sewage sludge was 50% lower than that in the presence of a mineralfertilizer.After one year, the authors found that both sewage sludge and mineral fertilizers were equally effective to improve P availability.Compost increased K availability by 11 and 20% after 1 and 3 years, which was attributed to the K supply by compost and the increase of exchange sites.The need for a longer-term experiment is stressed by these findings.When NPK is used alone but higher compared to the complete substitution by frass, the available K and P concentrations were the same.The 50NPK/50Frass treatment has a slightly slower K and P supply compared to other organic amendments, but this suggests that it reflects the rapid supply by the conventional fertilizer.The supply of organic carbon by frass might improve P availability by reducing sorption of P supplied by mineral fertilizer due to complexation with Fe, Al and Ca, which prevents P precipitation, and competition with inorganic P for the same sites42.The release of P and K from frass may be stimulated by the supply of mineral fertilizer, which is consistent with the elevated metabolic activity and diversity shown by this treatment.

Watersoluble P and available K and P are found in the soil.The values are average.At the 5% level, columns with the same letter don't differ much.

The 50NPK/50Frass treatment was as effective as the NPK treatment to improve barley bioaccumulation.N, K and P are contained in frass.The presence of N, K and P in a readily available form may be the reason for this.Nobile et al.23 reported that, due to high specific root length and great carboxylate release, barley was able to acquire as much P from sludge as from mineral fertilization.

The concentrations of N, K and P of the harvest.The values are average.At the 5% level, columns with the same letter don't differ much.

Compared to sole application of either NPK or frass, the combination of half doses of both did not have a significant impact on the plant.6).It is known that mixing organic amendment with mineral fertilizer will improve plant growth.The lack of synergistic effect between frass and NPK46 could be explained by the current pot experiment conditions where irrigation is controlled.In field experiment should be carried out to confirm the ability of frass to reduce the release of minerals.The effect of frass on NPK suggests that it could be used as a partial or complete substitute of mineral fertilization.Surprisingly, despite having much lower watersoluble P concentrations.Treatments with frass have the same effect as the NPK treatment.6).Watersoluble P is an important index for soil P status in the environment because it is related to P loss.Watersoluble P in NPK can be fixed in less available forms in the soil due to strong sorption.Since the P concentration in frass treatments was not impaired by the lower watersoluble P, it appears that P may be released during the incubation and become available for plant uptake and/or be protected by organic substances from sorption to sparingly available forms.frass prevents P from loss andsorption while acting as an effectivefertilizer by limiting watersoluble P concentration.One perspective of this work will be to test the effect of frass on the growth of other crop species in order to determine to what extent the ability to supply high amount of nutrients to plants is mediated by the strategies of plants.

In the next few years, insect production is expected to grow due to the increasing need for alternative sources of protein.It is necessary to take all components of the insects, including their frass, given the context of zero waste and the need to contribute to the circular economy.This study shows that frass can be used as a partial or complete substitute of mineral NPK.frass is similar to NPK in its ability to supply N, P and K and sustain the production of biomass.Watersoluble P concentration is up to five times lower in the presence of frass, which prevents P from loss and sorption onto soil.The presence of frass may increase the diversity and activity of the organisms in the soil.

Due to the fact that this was a greenhouse study, further in situ researches are required.The temporal release of nutrients for plants may be affected by the differences of temperature, soil, and crop.Our findings suggest that the growing amount of frass generated in the near future might be a sustainable resource for managing NPK nutrition.

Frass from mealworm was provided in the form of powder by nsect, an industrial company farming this insect at the large-scale.French and European regulations for farm animal feeds authorized the feeding of the mealworms exclusively on local agricultural raw materials.The frass was hygienized (70 C, 60 minutes) and used without any chemical input, making it compatible with organic farming and not subject to any specific restrictions.The temperature was 103 C.Water suspension (w/v), organic C content52, total nitrogen (N) content53, phosphorus (P), potassium (K), copper (Cu) and zinc (Zn) concentrations were analyzed for pH and electrical conductivity in a 1:5 frass.A modified Van Soest method55 was used to determine four biochemical fractions.

The frass particles were examined using a scanning electron microscope.The energy-dispersive X-ray spectrometer (EDS;Thermo Ultradry) was used to detect elements and element mapping was carried out on frass particles mounted on double-sided carbon tape.The count time was 200 seconds.

The soil in Beauvais was classified as a Haplic Luvisol56.A total mass of 100 kilograms was obtained from a cultivated land.The soil was air-dried, crushed and sieved.The soil was a silt loam with 16% sand, 67% silt, and 17% clay, according to the analysis done using the pipette method.The total N, organic C, and cation exchange capacity of the soil were revealed using the procedures described in Houben et al.58The available concentrations were Ca 3869, Mg 101, K 292, and P 72.

The frass was applied to the soil at a rate of 10 Mg dry matter.Adding the same quantity of N, P and K as in the Frass treatment was achieved by mixing the soil with appropriate amount of NPK.Adding 50% frass plus 50% NPK was also performed.

The frass treatments were followed in the laboratory with the French normalized59.The 25 g of dry soil mixture was put into 1.2 L hermetic jars and kept in a dark room.The experiment was done in four replicates.Demineralized water was used to control the water content of the mixture.After 1, 3, 7, 14, 21, 28, 49, 70 and 91 days, the CO2 was removed from each jar.The residual NaOH was used to determine the C-CO2 trapped in it.Incubation was done for N.Mineral N (N-NH4+ and N-NO3) was obtained by shaking the mixture for 1 h with 100 mL 1 mol L1 KCl after 60 days.A continuous flow analyser was used to analyse the mineral N in the extracts.The dynamics of C mineralisation in soil were calculated by subtracting C-CO2 mineralised from the control treatment and the results were expressed as a percentage of the total organic C.The release of mineral N from frass was calculated by subtracting the mineral that was measured in the control treatment from the one that had been treated.

A pot experiment was done to find out the effect of frass on plants.In four replicates, the plastic plant pots were filled with 3500 g of each mixture.The pots were placed in a dark room and the mixture was equilibrated for two weeks.The pots were placed in a greenhouse glass according to a randomized design after the equilibration period.There were 8 seeds sown in each pot.The plants were trimmed to two per pot after 10 days.The trials were conducted under controlled greenhouse conditions with daily sprinkler watering to keep the soil moist.After 9 weeks, shoots were dried and weighed and crushed.After aqua regia digestion, the content of P and K in aerial parts was analysed.The Dumas combustion method was used to analyse the content of N in aerial parts.

The water-soluble phosphorus concentration in the soil was determined after the harvest.The K and P concentrations were determined using a soil test.The soil pH was measured in H2O.Community-level physiological profiles were assessed using BIOLOG EcoPlates.Each 96-well plate had three replicates and 31 sole C sources.Five grams of soil collected at the end of the experiment were shaken with sterile 0.85% NaCl for 30 min at 200rpm and thenDiluted to 1: 1000.Each plate well was inoculated with 150 L of the dilution and the plates were incu bated at 25C.The color development for each well was obtained using an automated plate reader.Average well colour development is a measure of functional diversity.AWCD was calculated.

R and Ci are the absorbances of the control well and the plate well inoculated with C source.The Shannon-Weaver index was calculated.

After 72 h, the wells with the most active communities reached the asymptote of color development.The optimal time for further statistical analyses was suggested by Doan et al.62.

The data was analysed using descriptive statistics and normality was determined using a test.The data was subjected to one-way ANOVA and Tukey's post-hoc test to compare treatments.The data was subjected to two tests: the Kruskall-Wallis test and the Mann-Whitney test.R software version 3.5.063 and the package Rcmdr64 were used for the statistical analyses.

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The students of ASET 158 were thanked for technical assistance.Sébastien Potel is appreciated for his help.

D.H, G.D., M-P.F.And A-M.D.The experiments were conceived and designed.D.H. and A-M.D.The data was analyzed.The manuscript was written by D.H. with input from G.D., M-P. F.The manuscript was reviewed by all the authors.

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D. Houben, G., Faucon, MP.It was et al.Impact on crop growth and soil properties is a potential use of mealworm frass.The Sci Rep 10 will be published in 2020.The article is titled "S41598-020-61765-x."

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