Larvae of the black soldier fly (BSF) Hermetia illucens are polyphagous feeders and show tremendous bioconversion capabilities of organic matter into high-quality insect biomass. However, the digestion of lignocellulose-rich palm oil side streams such as palm kernel meal (PKM) is a particular challenge, as these compounds are exceptionally stable and are mainly degraded by microbes. This study aimed to investigate the suitability of BSF larvae as bioconversion agents of PKM. Since the intestinal microbiota is considered to play a key role in dietary breakdown and in increasing digestibility, the bacterial and fungal communities of BSF larvae were characterized in a culture-dependent approach and screened for their putative entomopathogenicity. The lethality of six putative candidates was investigated using intracoelomal injection. In total, 93 isolates were obtained with a bacterial share of 74% that were assigned to the four phyla Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. Members of the genera Klebsiella, Enterococcus, and Sphingobacterium are part of the core microbiome, as they were frequently described in the gut of Hermetia larvae regardless of diet, nutritional composition, or rearing conditions. With 75%, a majority of the fungal isolates belonged to the phylum Ascomycota. We identified several taxa already published to be able to degrade lignocelluloses, including Enterococcus, Cellulomonas, Pichia yeasts, or filamentous Fusarium species. The injection assays revealed pronounced differences in pathogenicity against the larvae. While Alcaligenes faecalis caused no, Diutina rugosa weak (23.3%), Microbacterium thalassium moderate (53.3%), and Pseudomonas aeruginosa and Klebsiella pneumoniae high (≥80%) lethality, Fusarium solani injection resulted in 100% lethality.

Insects offer a promising alternative source of protein to mitigate the environmental consequences of conventional livestock farming. Larvae of the black soldier fly (Hermetia illucens; Linnaeus, 1758) efficiently convert a variety of organic side streams and residues into valuable proteins, lipids, and chitin. Here, we evaluated the suitability of two palm oil industry side streams—empty fruit bunches (EFB) and palm kernel meal (PKM)—as larval feed, and their impact on the larval gut microbiome. Among 69 fungal species we screened, Marasmius palmivorus, Irpex consors, and Bjerkandera adusta achieved the fastest growth and lignin degradation, so these fungi were used for the pretreatment of 7:3 mixtures of EFB and PKM. Larvae reared on the mixture pretreated with B. adusta (BAD) developed significantly more quickly and reached a higher final weight than those reared on the other pretreatments or the non-fermented reference (NFR). Amplicon sequencing of the BAD and NFR groups revealed major differences in the larval gut microbiome. The NFR group was dominated by facultatively anaerobic Enterobacteriaceae (typical of H. illucens larvae) whereas the BAD group favored obligately anaerobic, cellulolytic bacteria (Ruminococcaceae and Lachnospiraceae). We hypothesize that fungal lignin degradation led to an accumulation of mycelia and subsequent cellulolytic breakdown of fiber residues, thus improving substrate digestibility.

The black soldier fly, Hermetia illucens (BSF; Diptera: Stratiomyidae), has come into the focus of research over the past decade since its larvae are polyphagous feeders with an exceptional substrate range, making them a promising candidate for the bioconversion of various organic side streams into valuable insect protein. While larval nutritional requirements have been studied in detail, basic information on adult feeding is still lacking. The reproduction of adult flies is a bottleneck and key determinant in rearing BSF, which has extensive potential for improvement. In the present study, we examined the impact of different carbohydrate (honey and d-glucose) and protein sources (Spirulina and Chlorella powder) on a variety of life history traits using a highly standardized single pair approach. Feeding a 5% honey solution was shown to make females live 2.8 d longer, become more fecund (9 egg clutches per 10 females), lay more eggs (increasing 1.7-fold to 182.4 mg per 10 females), reduce the number of failed oviposition events 3-fold and increase multiple oviposition events from 2 to 15. Additionally, female longevity after oviposition improved 1.7-fold from 6.7 to 11.5 d. In order to further optimize adult feeding, mixtures of proteins and carbohydrates with varying ratios should be tested.

The research on black soldier fly (BSF; Hermetia illucens L.; Diptera: Stratiomyidae) rearing is on the rise. The larval ability to grow on organic substances makes it an ideal candidate for the bioconversion of agricultural and other organic side streams. While there are several publications on the variables influencing the growth and development of different stages of BSF, juxtaposing the results could be amiss. This is because of the different experimental approaches and units used by the researchers. A few publications also lack information that might be necessary for comparing the results when using similar substrate and rearing conditions. In this review, we have analyzed the studies on rearing variables such as the type of feeding substrate, substrate depth and aeration, substrate temperature, substrate moisture, pH, feeding rate, and larval density mainly, but not exclusively, for the larvae. For the adults, factors such as the cage size, fly density, light, ambient temperature, and relative humidity are considered. In addition, larval performance when fed with side streams is encapsulated. This provides a backbone for future researchers to identify the already assessed variables along with their range and encourages them to define and use standardized rearing practices for a better comparison of the results.

Lipocalin 2 (LCN2) mediates key roles in innate immune responses. It has affinity for many lipophilic ligands and binds various siderophores, thereby limiting bacterial growth by iron sequestration. Furthermore, LCN2 protects against obesity and metabolic syndrome by interfering with the composition of gut microbiota. Consequently, complete or hepatocyte-specific ablation of the Lcn2 gene is associated with higher susceptibility to bacterial infections. In the present study, we comparatively profiled microbiota in fecal samples of wild type and Lcn2 null mice and show, in contrast to previous reports, that the quantity of DNA in feces of Lcn2 null mice is significantly lower than that in wild type mice (p < 0.001). By using the hypervariable V4 region of the 16S rDNA gene and Next-Generation Sequencing methods, we found a statistically significant change in 16 taxonomic units in Lcn2^-/- mice, including eight gender-specific deviations. In particular, members of Clostridium, Escherichia, Helicobacter, Lactococcus, Prevotellaceae_UCG-001 and Staphylococcus appeared to expand in the intestinal tract of knockout mice. Interestingly, the proportion of Escherichia (200-fold) and Staphylococcus (10-fold) as well as the abundance of intestinal bacteria encoding the LCN2-sensitive siderphore enterobactin (entA) was significantly increased in male Lcn2 null mice (743-fold, p < 0.001). This was accompanied by significant higher immune cell infiltration in the ileum as demonstrated by increased immunoreactivity against the pan-leukocyte protein CD45, the lymphocyte transcription factor MUM-1/IRF4, and the macrophage antigen CD68/Macrosialin. In addition, we found a higher expression of mucosal mast cell proteases indicating a higher number of those innate immune cells. Finally, the ileum of Lcn2 null mice displayed a high abundance of segmented filamentous bacteria, which are intimately associated with the mucosal cell layer, provoking epithelial antimicrobial responses and affecting T-helper cell polarization.

Rearing black soldier fly larvae (BSFL) in order to utilize biogenic materials is of increasing interest in the context of sustainable animal production. However, little is known about the amino acid (AA) requirements of this animal species. Therefore, a feeding experiment with BSFL was conducted, in which lysine, methionine, cysteine, arginine, phenylalanine, or histidine was reduced by 65% or methionine was increased by 65%. Reductions in single AAs in the substrate did not negatively impact BSFL growth, while the addition of methionine improved the growth performance, indicating that methionine was a limiting nutritional factor for maximal productivity. Differences in dietary AA profiles had no impact on the AA profile of the BSFL biomass. However, balance calculations for individual AAs indicated that the substrate microbiome was capable of synthesizing and balancing those AAs for microbial protein synthesis. While both the BSFL and microbiome utilized all free AAs, suggesting that supplemental AAs can be effective additives in BSFL production, this microbial AA synthesis avoided the negative performance impacts of BSFL provoked by severe AA reductions. The quantification of these effects suggested that the microbiome could add up to 35% to the overall AA supply in order to overcome AA deficiency. This effect may not necessarily ensure maximal BSFL productivity, as demonstrated by the extra addition of methionine to the substrate. Our research indicated that BSFL nutrition should consider interactions between substrate composition and microbial activity.