To treat or not to treat? Impact of hygienization on the microbiological safety of frass from black soldier fly larvae and yellow mealworm production aimed for fertilizer use

Ann De Volder , Lotte Frooninckx , David Deruytter , Johan Ceusters , Dries Vandeweyer

Bioresources and Bioprocessing ›› 2026, Vol. 13 ›› Issue (1) : 64

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Bioresources and Bioprocessing ›› 2026, Vol. 13 ›› Issue (1) :64 DOI: 10.1186/s40643-026-01030-7
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To treat or not to treat? Impact of hygienization on the microbiological safety of frass from black soldier fly larvae and yellow mealworm production aimed for fertilizer use
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Insect rearing residue / Hermetia illucens / Tenebrio molitor / Hygienization / Salmonella / E. coli / Enterococaceae

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Ann De Volder, Lotte Frooninckx, David Deruytter, Johan Ceusters, Dries Vandeweyer. To treat or not to treat? Impact of hygienization on the microbiological safety of frass from black soldier fly larvae and yellow mealworm production aimed for fertilizer use. Bioresources and Bioprocessing, 2026, 13(1): 64 DOI:10.1186/s40643-026-01030-7

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References

[1]

Amorim HCS, Ashworth AJ, Arsi K, et al.. Insect frass composition and potential use as an organic fertilizer in circular economies. J Econ Entomol, 2024, 117: 1261-1268

[2]

Balamurugan S, Ahmed R, Gao A. Survival of shiga toxin-producing Escherichia coli in broth as influenced by pH, water activity and temperature. Lett Appl Microbiol, 2014, 60: 341-346

[3]

Barragán-Fonseca KY, Nurfikari A, van de Zande EM, et al.. Insect frass and exuviae to promote plant growth and health. Trends Plant Sci, 2022, 27: 646-654

[4]

Beesigamukama D, Tanga CM, Sevgan S, et al.. Waste to value: global perspective on the impact of entomocomposting on environmental health, greenhouse gas mitigation and soil bioremediation. Sci Total Environ, 2023

[5]

Biswas S, Nazmi A, Pitesky M, et al.. Thermal inactivation of Escherichia coli and Salmonella typhimurium in poultry carcass and litter at thermophilic temperatures. J Appl Poult Res, 2019, 28: 307-317

[6]

Boltz TP, Boney JW, Shen C, et al.. The effect of standard pelleting and more thermally aggressive pelleting utilizing a hygieniser on feed manufacture and reduction of Enterococcus faecium, a Salmonella surrogate. J Appl Poult Res, 2019, 28: 1226-1233

[7]

Byappanahalli MN, Nevers MB, Korajkic A, et al.. Enterococci in the environment. Microbiol Mol Biol Rev, 2012, 76: 685-706

[8]

Cesaro C, Mannozzi C, Lepre A, et al.. Fate of Escherichia coli artificially inoculated in Tenebrio molitor L. larvae rearing chain for human consumption. Food Res Int, 2022, 157 Article ID: 111269

[9]

Chavez M, Uchanski M. Insect left-over substrate as plant fertiliser. J Insects Food Feed, 2021, 7: 683-694

[10]

Chen Z, Jiang X (2014) Microbiological safety of chicken litter or chicken litter-based organic fertilizers: a review. Agriculture (Switzerland) 4. https://doi.org/10.3390/agriculture4010001

[11]

Chen Z, Jiang X. Microbiological safety of animal wastes processed by physical heat treatment: an alternative to eliminate human pathogens in biological soil amendments as recommended by the Food Safety Modernization Act. J Food Prot, 2017, 80: 392-405

[12]

Coudron C, Deleu L, et al (2023) Kweekhandleiding Zwarte soldatenvlieg Hermetia illucens. Available at: https://inagro.be. Accessed Sept 2025

[13]

Cox NA, Burdick D, Bailey JS, Thomson JE. Effect of the steam conditioning and pelleting process on the microbiology and quality of commercial-type poultry feeds. Poult Sci, 1986, 65: 704-709

[14]

De Smet J, Wynants E, Cos P, Van Campenhout L. Microbial community dynamics during rearing of black. Appl Environ Microbiol, 2018, 84: 1-17

[15]

De Smet J, Vandeweyer D, Van Moll L, et al.. Dynamics of *Salmonella* inoculated during rearing of black soldier fly larvae (*Hermetia illucens*). Food Res Int, 2021

[16]

De Volder A, De Smet J, Frooninckx L, et al.. Heat treatment and storage of frass from black soldier fly larvae and yellow mealworm production: compliance with EU regulation on microbiological quality and safety. Microbiol Open, 2025

[17]

Delmon C, Prorot A, Maftah C, Casellas-Français M (2021) Thermal and acidic pre-treatments applied to cow manure: effects on pathogenic bacteria persistence and on biogas production during thermophilic anaerobic digestion. Available at: http://uest.ntua.gr/thessaloniki2021. Accessed Sept 2025

[18]

Dijk R, van den Berg D, Beumer R, et al.Microbiologie van voedingsmiddelen: Methoden, principes en criteria, 20155th ednRotterdamMybusinessmedia b.v.

[19]

EC. European Commission Regulation (EU) No 142/2011 of 25 February 2011 implementing Regulation (EC) No 1069/2009 of the European Parliament and of the Council laying down health rules as regards animal by-products and derived products not intended for human consumption and implementing Council Directive 97/78/EC as regards certain samples and items exempt from veterinary checks at the border under that Directive. Off J Eur Union, 2011, L 54: 1-254

[20]

EC. European Commission Regulation (EC) No 2021/1925 of the European Parliament and of the Council of 5 November 2021 amending certain annexes to Regulation (EU) No 142/2011 as regards the requirements for placing on the market of certain insect products and the adaptation of a containment method. Off J Eur Union, 2021, L393: 4-8

[21]

EC (2025) European Commission Regulation (EU) No 2025/1377 of 15 July 2025 amending and correcting Regulation (EU) No 142/2011 as regards certain requirements for the placing on the market and imports of animal by-products and derived products not intended for human consumption. Official Journal of the European Union

[22]

Gärtling D, Schulz H. Compilation of black soldier fly frass analyses. J Soil Sci Plant Nutr, 2021

[23]

Ho TTK, Tra VT, Le TH, et al.. Compost to improve sustainable soil cultivation and crop productivity. Case Stud Chem Environ Eng, 2022

[24]

Houben D, Daoulas G, Faucon M-P, Dulaurent A-M. Potential use of mealworm frass as a fertilizer: impact on crop growth and soil properties. Sci Rep, 2020, 10(1): 10:1-19

[25]

Hubbart JA, Kellner E, Petersen F. A 22-Site comparison of land-use practices, E. coli and Enterococci concentrations. Int J Environ Res Public Health, 2022

[26]

IPIFF (2021) An overview of the European market of insects as feed. Available at: https://ipiff.org/wp-content/uploads/2021. Accessed Sept 2025

[27]

IPIFF (2023) Overview of the European insect feed market. Available at: https://ipiff.org/wp-content/uploads/2023. Accessed Sept 2025

[28]

Klammsteiner T, Turan V, Juárez MFD, et al (2020) Suitability of black soldier fly frass as soil amendment and implication for organic waste hygienization. Agronomy 10. https://doi.org/10.3390/agronomy10101578

[29]

Lemmens B, Ceulemans J, Elslander H, et al.. Microbiological safety of animal wastes processed by physical heat treatment: an alternative to eliminate human pathogens in biological soil amendments as recommended by the Food Safety Modernization Act. J Food Prot, 2017, 80: 392-405

[30]

Lemmens B, Ceulemans J, Elslander H, et al (2006) Beste Beschikbare Technieken (BBT) voor mestverwerking. Available at: http://www.emis.vito.be. Accessed Sept 2025

[31]

Lepesteur M. Human and livestock pathogens and their control during composting. Crit Rev Environ Sci Technol, 2022, 52: 1639-1683

[32]

Li E, Saleem F, Edge TA, Schellhorn HE (2021) Biological indicators for fecal pollution detection and source tracking: a review. Processes 9. https://doi.org/10.3390/pr9112058

[33]

Lin M, Wang A, Ren L, et al.. Challenges of pathogen inactivation in animal manure through anaerobic digestion: a short review. Bioengineered Austin, 2022, 13: 1149-1161

[34]

Liu S, Roopesh MS, Tang J, et al.. Recent development in low-moisture foods: microbial safety and thermal process. Food Res Int, 2022

[35]

Lopes IG, Yong JWH, Lalander C. Frass derived from black soldier fly larvae treatment of biodegradable wastes. A critical review and future perspectives. Waste Manag, 2022, 142: 65-76

[36]

Lopes IG, Lalander C, Vidotti RM, Vinnerås B (2020) Reduction of bacteria in relation to feeding regimes when treating aquaculture waste in fly larvae composting. Front Microbiol 11. https://doi.org/10.3389/fmicb.2020.01616

[37]

Manyi-Loh CE, Mamphweli SN, Meyer EL, et al.. An overview of the control of bacterial pathogens in cattle manure. Int J Environ Res Public Health, 2016

[38]

Melse RW, De Buisonjé Bsc FE (2020) Manure treatment and utilisation options. Available at: https://edepot.wur.nl/541798. Accessed Sept 2025

[39]

Miller C, Heringa S, Kim J, Jiang X. Analyzing indicator microorganisms, antibiotic resistant Escherichia coli, and regrowth potential of foodborne pathogens in various organic fertilizers. Foodborne Pathog Dis, 2013, 10: 520-527

[40]

Ongeng D, Geeraerd AH, Springael D, et al.. Fate of Escherichia coli O157:H7 and Salmonella enterica in the manure-amended soil-plant ecosystem of fresh vegetable crops: a review. Crit Rev Microbiol, 2015, 41: 273-294

[41]

Osimani A, Milanović V, Cardinali F, et al.. The bacterial biota of laboratory-reared edible mealworms (Tenebrio molitor L.): from feed to frass. Int J Food Microbiol, 2018, 272: 49-60

[42]

Osimani A, Milanović V, Roncolini A, et al.. Hermetia illucens in diets for zebrafish (Danio rerio): a study of bacterial diversity by using PCR-DGGE and metagenomic sequencing. PLoS ONE, 2019

[43]

OVAM (2022) Onderzoek “Frass” Restsubstraat van Insectenkweek (Besteknummer 8278). Available at: https://www.vlaanderen.be/publicaties. Accessed Sept 2025.

[44]

Pourcher A-M, Burton C, Ziebal C, De-Guardia A (2009) Impact of temperature-time combinations on enteric bacteria in separated solids from pig manure. Available at: https://ramiran.uvlf.sk/doc08. Accessed Sept 2025

[45]

Poveda J. Insect frass in the development of sustainable agriculture. A review. Agron Sustain Dev, 2021, 41 Article ID: 5

[46]

Poveda J, Jiménez-Gómez A, Saati-Santamaría Z, et al.. Mealworm frass as a potential biofertilizer and abiotic stress tolerance-inductor in plants. Appl Soil Ecol, 2019, 142: 110-122

[47]

Praeg N, Klammsteiner T. Primary study on frass fertilizers from mass-reared insects: species variation, heat treatment effects, and implications for soil application at laboratory scale. J Environ Manage, 2024, 356 Article ID: 120622

[48]

Rolfe C, Daryaei H (2020) Intrinsic and extrinsic factors affecting microbial growth in food systems. In: Food Engineering Series. Springer, pp 3–24. https://doi.org/10.1007/978-3-030-42660-6_1

[49]

Sharara AM, Aguirre-Villegas AH, Larson AR, et al (2021) Pelleting Animal Manures. Available at: https://content.ces.ncsu.edu. Accessed Sept 2025

[50]

Siddiqui SA, Gadge AS, Hasan M, et al.. Future opportunities for products derived from black soldier fly (BSF) treatment as animal feed and fertilizer—a systematic review. Environ Dev Sustain, 2024

[51]

Susinchain Sustaineble Insect Chain (2022) Deliverable 6.2 Chemical and microbiological safety of insect frass. Available at: https://susinchain.eu/wp-content/uploads/2023. Accessed Sept 2025

[52]

Swinscoe I, Oliver DM, Gilburn AS, et al.. Seaweed-fed black soldier fly (Hermetia illucens) larvae as feed for salmon aquaculture: Assessing the risks of pathogen transfer. J Insects Food Feed, 2019, 5: 15-27

[53]

Tontti T, Heinonen-Tanski H, Karinen P, et al.. Maturity and hygiene quality of composts and hygiene indicators in agricultural soil fertilised with municipal waste or manure compost. Waste Manag Res, 2011, 29: 197-207

[54]

UK Health Security Agency (2024) Guidelines for Assessing the Microbiological Safety of Ready-to-Eat Foods Placed on the Market. Available at: https://assets.publishing.service.gov.uk/media. Accessed Sept 2025

[55]

Van Looveren N, Vandeweyer D, van Schelt J, Van Campenhout L. Occurrence of Clostridium perfringens vegetative cells and spores throughout an industrial production process of black soldier fly larvae (Hermetia illucens). J Insects Food Feed, 2021, 2011: 1-10

[56]

Van Looveren N, Vandeweyer D, Van Campenhout L. Impact of heat treatment on the microbiological quality of frass originating from black soldier fly larvae. InSects, 2022, 13: 1-15

[57]

Van Looveren N, IJdema F, van der Heijden N, et al.. Microbial dynamics and vertical transmission of Escherichia coli across consecutive life stages of the black soldier fly (Hermetia illucens). Anim Microbiome, 2024

[58]

Vandeweyer D, Lievens B, Van Campenhout L. Identification of bacterial endospores and targeted detection of foodborne viruses in industrially reared insects for food. Nat Food, 2020, 1: 511-516

[59]

Vandeweyer D, Bruno D, Bonelli M, et al.. Bacterial biota composition in gut regions of black soldier fly larvae reared on industrial residual streams: revealing community dynamics along its intestinal tract. Front Microbiol, 2023

[60]

Wynants E, Frooninckx L, Crauwels S, et al.. Assessing the microbiota of black soldier fly larvae (Hermetia illucens) reared on organic waste streams on four different locations at laboratory and large scale. Microb Ecol, 2019, 77: 913-930

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VLAIO-TETRA ValoReSect Project(HBC.2021.0103)

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