Hemolymph metabolism of black soldier fly (Diptera: Stratiomyidae), response to different supplemental fungi.

The black soldier fly, Hermetia illucens L. (Diptera: Stratiomyidae), is commonly used for organic waste recycling and animal feed production. However, the often inadequate nutrients in organic waste necessitate nutritional enhancement of black soldier fly larvae, e.g., by fungal supplementation of its diet. We investigated the amino acid composition of two fungi, Candida tropicalis (Castell.) Berkhout (Saccharomycetales: Saccharomycetaceae) and Pichia kudriavzevii Boidin, Pignal & Besson (Saccharomycetales: Pichiaceae), from the black soldier fly gut, and commercial baker's yeast, Saccharomyces cerevisiae Meyen ex E.C. Hansen (Saccharomycetales: Saccharomycetaceae), and their effects on larval growth and hemolymph metabolites in fifth-instar black soldier fly larvae. Liquid chromatography-mass spectrometry was used to study the effect of fungal metabolites on black soldier fly larval metabolism. Amino acid analysis revealed significant variation among the fungi. Fungal supplementation led to increased larval body mass and differential metabolite accumulation. The three fungal species caused distinct metabolic changes, with each over-accumulating and down-accumulating various metabolites. We identified significant alteration of histidine metabolism, aminoacyl-tRNA biosynthesis, and glycerophospholipid metabolism in BSF larvae treated with C. tropicalis. Treatment with P. kudriavzevii affected histidine metabolism and citrate cycle metabolites, while both P. kudriavzevii and S. cerevisiae treatments impacted tyrosine metabolism. Treatment with S. cerevisiae resulted in down-accumulation of metabolites related to glycine, serine, and threonine metabolism. This study suggests that adding fungi to the larval diet significantly affects black soldier fly larval metabolomics. Further research is needed to understand how individual amino acids and their metabolites contributed by fungi affect black soldier fly larval physiology, growth, and development, to elucidate the interaction between fungal nutrients and black soldier fly physiology.

Effect of entomopathogenic fungus Beauveria bassiana on the growth characteristics and metabolism of black soldier fly larvae.

Beauveria bassiana is an entomopathogenic fungus widely used in agriculture to reduce populations of various pests. However, when agricultural waste is utilized for organic recycling, B. bassiana has the potential to impact recycling performance, by affecting the survival, and body mass of decomposing organisms (such as insect's larvae). Additionally, in natural conditions where decayed organic matter contains a high load of different entomopathogenic organisms, larval growth may be affected when consumed or in contact. In a laboratory study, we aimed to comprehend the effects of B. bassiana on the growth characteristics and larval metabolism of the black soldier fly larvae, which is a known decomposing insect. The experiments used both feeding (mixing the spores with the diet, hereafter BF) and contact treatments (by dipping the larva in the spores solution, hereafter BD), and were compared to a water-treated control group. The BF treatment significantly reduced larval body weight, adult emergence, and adult weight compared to both the control and the BD treatment. Furthermore, an analysis of hemolymph metabolites, categorized by class, indicated a higher accumulation of metabolites belonging to the purine and purine derivative classes, as well as carboxylic acids and their derivatives, including peptides and oligopeptides, indicating potential disruption of protein synthesis or degradation caused by the BF treatment. Pathway enrichment analysis showed significant alterations in purine metabolism and D-Arginine and D-ornithine metabolism compared to the control. Taurine and hypotaurine metabolism were significantly altered in the BD treatment compared to the control but not significantly enriched in the BF treatment. Our results suggest that the BF treatment impairs protein synthesis or degradation, affecting larval growth characteristics. Future studies should explore innate immunity-related gene expression and antimicrobial peptide production in BSF larvae to understand their immunity to pathogens.

Assessing Fungal Diversity and Abundance in the Black Soldier Fly and its Environment.

Detritivorous insects that flourish in decaying environments encounter microorganisms throughout their life cycle. However, it is not clear whether the microbial composition of the decaying environment affects the microbial composition of the insect gut, or whether the opposite is true, with the microorganisms that are adapted to the insect's digestive system being dispersed by the insects to new habitats, thereby becoming more and more common in the environment. To test these questions the fungal composition of the black soldier fly (BSF) (Stratiomyidae; Hermetia illucens Linnaeus) larval gut and its surrounding decaying environment (household compost bins) were analyzed using amplicon sequencing. Constancy in the dominance of the genus Candida (Debaryomycetaceae) in most of the environments and larval guts was found. This finding may suggest a 'core' structure to the fungal community of the BSF. In locations where nutrient composition of the environment had higher fiber content, the Candida was not dominant and the most common fungi were the genus Gibberella (Nectriaceae) and the family Dipodascaceae. The later was dominant also in the larval gut and the former was replaced with Meyerozyma (Debaryomycetaceae), which may suggest a selection process by the insect's gut. Little is known about the ecological interactions of insects with eukaryotic microorganisms, such as yeast-like fungi. As their metabolic complexity and ability is intense, they have the potential to dramatically affect the physiological condition of the insect.

Does Consumption of Baker's Yeast (Saccharomyces cerevisiae) by Black Soldier Fly (Diptera: Stratiomyidae) Larvae Affect Their Fatty Acid Composition?

Fatty acids are important compounds for insects, but the requirements for essential fatty acids may differ between insect species. Most of the fatty acids are acquired through the insect's diet; therefore, supplementing the diet with baker's yeast (Saccharomyces cerevisiae Meyen ex E.C. Hansen), which produces unsaturated fatty acids, was predicted to affect the fatty acid composition of the insect. The tested insect was the black soldier fly (BSF) (Hermetia illucens L.), that is used as a source of protein and fat in feed. Therefore, there is importance for BSF larvae (BSFL) nutritional composition, especially the unsaturated fatty acids content, which is one of the nutritional limitations for mammalian diets. The dominant fatty acids of the tested BSFL were the saturated fatty acids: lauric, myristic, and palmitic acids, as found in other BSF studies. Oleic acid (c18:1) and linoleic acid (C18:2) were the abundant unsaturated fatty acids in the BSFL. The proportion of linoleic acid was higher in the substrate with the supplemental yeast; however, this did not affect its proportion in the larvae. The higher proportion of linoleic acid may have been exploited as a source for production of saturated lauric acid. Therefore, providing unsaturated fatty acids to the substrate through supplemental baker's yeast is not the most efficient way to increase the proportion of unsaturated fatty acids in the larvae.

Comparative untargeted metabolic analysis of natural- and laboratory-reared larvae of black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae).

In the present study, we examined the metabolic composition of black soldier fly (BSF) larvae from natural populations (Ruhama: R and She'ar Yashuv: S) and from a laboratory-reared colony (C) using untargeted metabolomics analysis. The results revealed significant over-accumulation of metabolites from phenylalanine and purine metabolism and biosynthesis of phenylalanine, tyrosine and tryptophan, and arginine in both natural populations, and enriched pathway analysis, compared to the laboratory-reared colony. In addition, we found accumulation of glutathione metabolism and aminoacyl tRNA biosynthesis related metabolites in R, and linoleic acid and tryptophan metabolism related metabolites in S. Moreover, we found down-accumulation of metabolites belonging to alanine, aspartate and glutamate metabolism in both natural populations: amino sugar and nucleotide sugar metabolism only in the R population and aminoacyl-tRNA biosynthesis, glyoxylate and dicarboxylate metabolism only in the S population. Overall, the results suggest that the naturally growing larvae require large quantities of metabolites from aromatic amino acids (phenylalanine, tyrosine and tryptophan) for defense against pathogens under natural conditions e.g., melanization. In addition, glutathione metabolites help the BSF to survive under oxidative stress. Further study of the functional metabolomics of naturally growing and laboratory-reared larvae could provide a platform for better understanding of BSF larval survival mechanisms in complex environments.