journal article·2026-02-11Open access

Study of Probiotic-Enriched Diet Effects on Yellow Mealworm Larvae Production at Laboratory and Pilot Scales

Sabina Dahal, Aberham Hailu Feyissa, Lucas Sales Queiroz, Antoine Lecocq, Heidi Amlund, Katrine Kastberg, Uri Lesmes, Federico Casanova

Applied Sciences· Volume 16· Issue 4

Abstract

Global food security is an increasing challenge due to population growth and the limited availability of natural resources, driving the search for sustainable protein sources. In this context, edible insects such as yellow mealworms (Tenebrio molitor) have emerged as a promising alternative, while probiotics have been widely applied in animal production to enhance growth performance and nutritional quality. This study aimed to evaluate the effects of probiotic supplementation on the growth performance, biomass yield, and nutritional composition of yellow mealworm larvae at laboratory and pilot scales. Three probiotic strains—Bacillus velezensis, Bacillus coagulans, and Pediococcus pentosaceus—were tested at four different dosage levels, using wheat bran and brewer’s spent grain as feed substrates. Larval growth was monitored weekly, and total harvested biomass, proximate composition (dry matter, protein, fat, and ash), amino acid profile, and mineral composition were determined using standardized analytical methods. At the laboratory scale, probiotic supplementation did not result in significant differences in mean larval weight or total biomass (p > 0.05). In contrast, at the pilot scale, significant improvements in larval growth and biomass were observed for specific probiotic treatments, with mean larval weights reaching approximately 140–150 mg and total harvest biomass increases of up to ~15% compared to the control (p < 0.05). Growth curves at both scales followed a sigmoidal pattern with a high correlation between laboratory and pilot experiments (R2 = 0.98). Probiotic supplementation did not significantly affect crude protein content, but alterations in fat content, specific amino acid concentrations, and mineral composition were observed at the pilot scale, depending on strain and dosage. Overall, the results demonstrate that probiotic supplementation can enhance yellow mealworm production under pilot-scale conditions, while laboratory-scale trials may not fully capture these effects. These findings highlight the importance of scale when evaluating probiotic strategies and support the potential application of Bacilli-based probiotics to improve the efficiency and nutritional quality of industrial insect production systems.
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Research context

OpenAlex topics: Insect Utilization and Effects · Invertebrate Immune Response Mechanisms · Aquaculture Nutrition and Growth

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Food Production Engineering Research Group (FPE), National Food Institute, Technical University of Denmark (DTU), 2800 Kongens Lyngby, Denmark

Section of Organismal Biology, Department of Plant and Environmental Sciences (PLEN), University of Copenhagen, 1172 Frederiksberg, Denmark

Research Group for Analytical Food Chemistry, National Food Institute, Technical University of Denmark (DTU), 2800 Kongens Lyngby, Denmark

Lab of Food Chemistry, Bioactives and Digestion, Faculty of Biotechnology and Food Engineering, Technion-Israel Institute of Technology, Haifa 3200003, Israel

References (74)

Show source-provided references
1. United Nations (2019). World population prospects 2019: Highlights. World Population Prospects, United Nations.
2. Potential of insects as food and feed in assuring food security10.1146/annurev-ento-120811-153704 ↗
3. Nutritional composition and safety aspects of edible insects10.1002/mnfr.201200735 ↗
4. Oonincx, D.G.A.B., and de Boer, I.J.M. (2012). Environmental impact of the production of mealworms as a protein source for humans—A Life Cycle Assessment. PLoS ONE, 7.10.1371/journal.pone.0051145 ↗
5. Opportunities and hurdles of edible insects for food and feed10.1111/nbu.12291 ↗
6. Amelioration of growth, nutritional value, and microbial load of Tenebrio molitor (Coleoptera: Tenebrionidae) through probiotic supplemented feed10.1007/s00217-021-03925-5 ↗
7. Complete nutrient composition of commercially raised invertebrates used as food for insectivores10.1002/zoo.10031 ↗
8. Dahal, S., Jensen, A.B., and Lecocq, A. (2022). Effect of probiotics on Tenebrio molitor larval development and resistance against the fungal pathogen Metarhizium brunneum. Insects, 13.10.3390/insects13121114 ↗
9. Kooienga, E.M., Baugher, C., Currin, M., Tomberlin, J.K., and Jordan, H.R. (2020). Effects of bacterial supplementation on black soldier fly growth and development at benchtop and industrial scale. Front. Microbiol., 11.10.3389/fmicb.2020.587979 ↗
10. Gorrens, E., Lecocq, A., and De Smet, J. (2023). The use of probiotics during rearing of Hermetia illucens: Potential, caveats, and knowledge gaps. Microorganisms, 11.10.3390/microorganisms11020245 ↗
11. Guerfali, M.M., Hamden, H., Charaabi, K., Fadhl, S., Mosbah, A., and Cherif, A. (2023). Probiotics as a Beneficial Modulator of Gut Microbiota and Environmental Stress for Sustainable Mass-Reared Ceratitis Capitata, IntechOpen.
12. Savio, C., Herren, P., Rejasse, A., Rios, A., Bourelle, W., Bruun-Jensen, A., Lecocq, A., van Loon, J.J., and Nielsen-LeRoux, C. (2024). Minor impact of probiotic bacteria and egg white on Tenebrio molitor growth, microbial composition, and pathogen infection. Front. Insect Sci., 4.10.3389/finsc.2024.1334526 ↗
13. Pediococcus pentosaceus ZZ61 enhances growth performance and pathogenic resistance of silkworm Bombyx mori by regulating gut microbiota and metabolites10.1016/j.biortech.2024.130821 ↗
14. Won, S., Hamidoghli, A., Choi, W., Bae, J., Jang, W.J., Lee, S., and Bai, S.C. (2020). Evaluation of potential probiotics Bacillus subtilis WB60, Pediococcus pentosaceus, and Lactococcus lactis on growth performance, immune response, gut histology and immune-related genes in whiteleg shrimp, Litopenaeus vannamei. Microorganisms, 8.10.3390/microorganisms8020281 ↗
15. Yang, S., Gong, P., Pan, J., Wang, N., Tong, J., Wang, M., Long, M., Li, P., and He, J. (2019). Pediococcus pentosaceus xy46 can absorb zearalenone and alleviate its toxicity to the reproductive systems of male mice. Microorganisms, 7.10.3390/microorganisms7080266 ↗
16. Probiotic properties of an indigenous Pediococcus pentosaceus strain on Tenebrio molitor larval growth and survival10.3920/JIFF2020.0156 ↗
17. Iorizzo, M., Letizia, F., Ganassi, S., Testa, B., Petrarca, S., Albanese, G., Di Criscio, D., and De Cristofaro, A. (2022). Functional properties and antimicrobial activity from lactic acid bacteria as resources to improve the health and welfare of honeybees. Insects, 13.10.3390/insects13030308 ↗
18. Impact of density, reproduction period and age on fecundity of the yellow mealworm Tenebrio molitor (Coleoptera: Tenebrionidae)10.3920/JIFF2017.0013 ↗
19. Yellow mealworm: Effects of adults breeding density on adults and larvae performances from an industrial perspective10.1016/j.animal.2024.101360 ↗
20. Impact of adult weight, density, and age on reproduction of Tenebrio molitor (coleoptera: Tenebrionidae)
21. The effects of Carbon dioxide exposure concentrations on human vigilance and sentiment in an enclosed workplace environment10.1111/ina.12746 ↗
22. Chen, L., Lv, C., Li, B., Zhang, H., Ren, L., Zhang, Q., Zhang, X., Gao, J., Sun, C., and Hu, S. (2021). Effects of Bacillus velezensis supplementation on the growth performance, immune responses, and intestine microbiota of Litopenaeus vannamei. Front. Mar. Sci., 8.10.3389/fmars.2021.744281 ↗
23. Pei, Y., Zhao, S., Chen, X., Zhang, J., Ni, H., Sun, M., Lin, H., Liu, X., Chen, H., and Yang, S. (2022). Bacillus velezensis EEAM 10B strengthens nutrient metabolic process in black soldier fly larvae (Hermetia illucens) via changing gut microbiome and metabolic pathways. Front. Nutr., 9.10.3389/fnut.2022.880488 ↗
24. Ye, M., Wei, C., Khalid, A., Hu, Q., Yang, R., Dai, B., Cheng, H., and Wang, Z. (2020). Effect of Bacillus velezensis to substitute in-feed antibiotics on the production, blood biochemistry and egg quality indices of laying hens. BMC Vet. Res., 16.10.1186/s12917-020-02570-6 ↗
25. Effect of dietary probiotic, Bacillus coagulans, on growth performance, chemical composition, and meat quality of Guangxi Yellow chicken10.3382/ps.2009-00319 ↗
26. Dietary Bacillus velezensis T23 fermented products supplementation improves growth, hepatopancreas and intestine health of Litopenaeus vannamei10.1016/j.fsi.2024.109595 ↗
27. AOAC (1990). Official Methods of Analysis, Association of Official Analytical Chemist. [15th ed.].
28. Physico-chemical and colloidal properties of protein extracted from black soldier fly (Hermetia illucens) larvae10.1016/j.ijbiomac.2021.07.081 ↗
29. Protein extraction from yellow mealworm (Tenebrio molitor) assisted by pulsed electric fields: Effect on foaming properties10.1016/j.lwt.2024.117041 ↗
30. A comparison of growth curve models for forensically important flies using sigmoid functions10.7580/kjlm.2020.44.2.84 ↗
31. Effect of low and high concentrate supplementation on health and welfare indicators in different breeds in small-scale mountain dairy farms10.1017/S0022029921000273 ↗
32. Meier, L. (2022). ANOVA and Mixed Models: A Short Introduction Using R, Chapman and Hall/CRC. [1st ed.].10.1201/9781003146216 ↗
33. Yu, X., He, Q., and Wang, D. (2021). Dynamic analysis of major components in the different developmental stages of Tenebrio molitor. Front. Nutr., 8.10.3389/fnut.2021.689746 ↗
34. Testing mechanistic models of growth in insects10.1098/rspb.2015.1973 ↗
35. Effect of larval density on food utilization efficiency of Tenebrio molitor (Coleoptera: Tenebrionidae)10.1093/jee/tov208 ↗
36. Eberle, S., Schaden, L.M., Tintner, J., Stauffer, C., and Schebeck, M. (2022). Effect of temperature and photoperiod on development, survival, and growth rate of mealworms, Tenebrio molitor. Insects, 13.10.3390/insects13040321 ↗
37. Kopecká, A., Kouřimská, L., Škvorová, P., Kurečka, M., and Kulma, M. (2024). Effect of temperature on the nutritional quality and growth parameters of yellow mealworm (Tenebrio molitor L.): A Preliminary Study. Appl. Sci., 14.10.3390/app14062610 ↗
38. The Effect of relative humidity on the survival and growth rate of the yellow mealworm larvae (Tenebrio molitor, Linnaeus 1758)10.3920/JIFF2020.0068 ↗
39. CO2 effects on larval development and genetics of mealworm beetle, Tenebrio molitor L. (Coleoptera: Tenebrionidae) in two different CO2 systems10.15666/aeer/1602_17491766 ↗
40. The effect of elevated CO2 on life cycle of Tenebrio molitor L. (Coleoptera: Tenebrionidae)
41. Fungal secondary metabolites as modulators of interactions with insects and other arthropods10.1016/j.fgb.2010.08.008 ↗
42. Preservation techniques and their effect on nutritional values and microbial population of brewer’s spent grain: A review10.1186/s43170-022-00120-8 ↗
43. Li, J., Xu, L., Liao, J., Li, X., Wang, X., Wu, Q., and Zhu, J. (2023). In vitro intestinal simulation system on the carbon source utilization characteristics and mechanism of interspecific syntrophic effects of Bifidobacterium longum CECT7894 and Pediococcus pentosaceus CECT8330. Front. Pediatr., 11.10.3389/fped.2023.1276846 ↗
44. Raccach, M. (2014). Pediococcus. Encyclopedia of Food Microbiology, Elsevier Inc.. [2nd ed.].10.1016/B978-0-12-384730-0.00247-0 ↗
45. Competition with filamentous fungi and its implication for a gregarious lifestyle in insects living on ephemeral resources10.1111/j.0307-6946.2005.00722.x ↗
46. Estimate of chitin in raw whole insects10.1002/zoo.20123 ↗
47. Impact of elevated CO2 on tri-trophic interaction of Gossypium hirsutum, Aphis gossypii, and Leis axyridis10.1603/0046-225X-34.1.37 ↗
48. Growth performance and feed conversion efficiency of three edible mealworm species (Coleoptera: Tenebrionidae) on diets composed of organic by-products
49. Influence of larval density and dietary nutrient concentration on performance, body protein, and fat contents of black soldier fly larvae (Hermetia illucens)10.1111/eea.12716 ↗
50. Biotransformation of insect processing residues: Production of lactic acid bacterial biomass and associated partial removal of proteins from chitin10.1016/j.biortech.2024.131540 ↗
51. Comparison of sustainable lipid and protein removal methods for the isolation of insect chitin from black soldier fly exoskeleton10.1021/acsfoodscitech.0c00104 ↗
52. Chitin and chitosan preparation from marine sources. Structure, properties and applications10.3390/md13031133 ↗
53. Gatti, J.L., Lemauf, S., Belghazi, M., Arthaud, L., and Poirié, M. (2024). In Drosophila hemolymph, serine proteases are the major gelatinases and caseinases. Insects, 15.10.3390/insects15040234 ↗
54. Larvae of mealworm (Tenebrio molitor L.) as European novel food
55. Insight into the proximate composition and microbial diversity of edible insects marketed in the European Union10.1007/s00217-016-2828-4 ↗
56. Minerals in edible insects: A review of content and potential for sustainable sourcing
57. The variability of physico-chemical properties of brewery spent grain from 8 different breweries10.1016/j.heliyon.2021.e06583 ↗
58. Association between the gut microbiota and mineral metabolism10.1002/jsfa.8724 ↗
59. Probiotic-driven advancement: Exploring the intricacies of mineral absorption in the human body
60. The impact of probiotics on gut health via alternation of immune status of monogastric animals10.1016/j.aninu.2020.11.004 ↗
61. Bioaccumulation of heavy metals in the black soldier fly, Hermetia illucens and effects on its life cycle10.3920/JIFF2015.0030 ↗
62. Biofortification of selenium in black soldier fly (Hermetia illucens) prepupae reared on seaweed or selenium enriched substrates10.3920/JIFF2021.0153 ↗
63. Advances in selenium supplementation: From selenium-enriched yeast to potential selenium-enriched insects, and selenium nanoparticles10.1016/j.aninu.2023.05.002 ↗
64. Bioaccumulation of cadmium, lead, and zinc in agriculture-based insect food chains10.1007/s10661-018-7051-2 ↗
65. Bioaccumulation of copper, zinc, cadmium and lead by Bacillus sp., Bacillus cereus, Bacillus sphaericus and Bacillus subtilis
66. Beglari, S., Fereshteh, S., Milani, M., and Rezaie, N. (2025, March 27). Title Page 2 Lactoremediation: Heavy Metals Elimination from the Gastrointestinal Tract by Lactic acid Bacteria. Available online: https://www.sciencedirect.com/science/article/pii/S2212429223008532.
67. Effect of probiotics on the chemical, mineral, and amino acid composition of broiler chicken meat10.15421/20178 ↗
68. Regulation of probiotics on metabolism of dietary protein in intestine10.2174/1389203720666191111112941 ↗
69. Meyer-Rochow, V.B., Gahukar, R.T., Ghosh, S., and Jung, C. (2021). Chemical composition, nutrient quality and acceptability of edible insects are affected by species, developmental stage, gender, diet, and processing method. Foods, 10.10.3390/foods10051036 ↗
70. Extraction and characterisation of protein fractions from five insect species10.1016/j.foodchem.2013.05.115 ↗
71. Comparison of larval and pupal cuticular proteins in Tenebrio molitor10.1016/0965-1748(94)00048-M ↗
72. A chymotrypsin-like proteinase from the midgut of Tenebrio molitor larvae10.1016/j.biochi.2005.02.013 ↗
73. Protein identification and in vitro digestion of fractions from Tenebrio molitor10.1007/s00217-015-2632-6 ↗
74. Influence of the processing on composition, protein structure and techno-functional properties of mealworm protein concentrates produced by isoelectric precipitation and ultrafiltration/diafiltration10.1016/j.foodchem.2024.139177 ↗