Pupa

In social groups, infections have the potential to spread rapidly and cause disease outbreaks. Here, we show that in a social insect, the ant Lasius neglectus, the negative consequences of fungal infections (Metarhizium brunneum) can be mitigated by employing an efficient multicomponent behaviour, termed destructive disinfection, which prevents further spread of the disease through the colony. Ants specifically target infected pupae during the pathogen's non-contagious incubation period, utilising chemical 'sickness cues' emitted by pupae. They then remove the pupal cocoon, perforate its cuticle and administer antimicrobial poison, which enters the body and prevents pathogen replication from the inside out. Like the immune system of a metazoan body that specifically targets and eliminates infected cells, ants destroy infected brood to stop the pathogen completing its lifecycle, thus protecting the rest of the colony. Hence, in an analogous fashion, the same principles of disease defence apply at different levels of biological organisation.

Although increasingly targeted in animal nutrition, black soldier fly larvae or prepupae (BSF, Hermetia illucens L. 1758) require the characterization and modulation of their fatty acid profile to become fully integrated within the feed sector. This improvement will only be possible by the understanding of underlaying biochemical pathways of fatty acid synthesis in BSF. In this study, we hypothesized a labelling of de novo synthesized fatty acids in BSF by the incorporation of deuterated water (D2O) in their feed. Three batches of fifty larvae were reared on two diets with different polyunsaturated fatty acid profiles moistened with 40% of H2O or D2O: chicken feed or 40% of chicken feed and 60% of flax cake. Although the occurrence of D2O in insect feed increased the larval development time and decreased prepupal weight, it was possible to track the biosynthesis of fatty acids through deuterium labelling. Some fatty acids (decanoic, lauric or myristic acid) were exclusively present in their deuterated form while others (palmitic, palmitoleic or oleic acid) were found in two forms (deuterated or not) indicating that BSF can partially produce these fatty acids via biosynthesis pathways and not only by bioaccumulation from the diet. These results suggest the importance of carbohydrates as a source of acetyl-CoA in the constitution of the BSF fatty acid profile but also the potential importance of specific enzymes (e.g. thioesterase II or Δ12 fat2 desaturase) in BSF fatty acid metabolism. Finally, nearly no deuterated polyunsaturated fatty acids were found in BSF fed with deuterium confirming that BSF is not able to produce these types of fatty acids. Despite the high levels of linolenic acid in flax-enriched diets, BSF will simply bioaccumulate around 13% of this fatty acid and will metabolize approximately two-thirds of it into saturated fatty acids as lauric or myristic acid.

Deformed wing virus (DWV) is an emerging infectious disease of the honey bee (Apis mellifera) that is considered a major cause of elevated losses of honey bee colonies. DWV comprises two widespread genotypes: the originally described genotype A, and genotype B. In adult honey bees, DWV-B has been shown to be more virulent than DWV-A. However, their comparative effects on earlier host developmental stages are unknown. Here, we experimentally inoculated honey bee pupae and tested for the relative impact of DWV-A versus DWV-B on mortality and wing deformities in eclosing adults. DWV-A and DWV-B caused similar, and only slightly elevated, pupal mortality (mean 18% greater mortality than control). Both genotypes caused similarly high wing deformities in eclosing adults (mean 60% greater wing deformities than control). Viral titer was high in all of the experimentally inoculated eclosing adults, and was independent of wing deformities, suggesting that the phenotype 'deformed wings' is not directly related to viral titer or viral genotype. These viral traits favor the emergence of both genotypes of DWV by not limiting the reproduction of its vector, the ectoparasitic Varroa destructor mite, in infected pupae, and thereby facilitating the spread of DWV in honey bees infested by the mite.

Bats are important reservoirs of zoonotic viruses, yet the role of their obligate ectoparasites, particularly bat flies, in viral maintenance and transmission remains poorly understood. Although viral sequences have been detected in adult bat flies, vertical transmission to their offspring has not been investigated. Here, we present the first metagenomic analysis of bat fly pupae to assess transgenerational viral persistence. Eighty pupae associated with a large Miniopterus schreibersii colony in Hungary were analyzed by next-generation sequencing. We generated more than 69 million reads and over 28,000 assembled contigs. Although initial classification suggested the presence of viral sequences, none were confirmed following sequence-level validation. These findings highlight the substantial potential for false-positive viral assignments in metagenomic datasets and the importance of rigorous post-classification validation. While our results provide no evidence that vertical transmission is a major mechanism of viral maintenance in bat-bat fly systems, this conclusion is limited by the relatively small sample size and single sampling occasion. Highly divergent or low-abundance viruses may also have remained undetected. Broader sampling across seasons and locations is therefore needed to assess the generality of these findings. Nevertheless, this study provides a valuable first step toward understanding potential transgenerational viral persistence in bat flies and highlights the need for complementary approaches to clarify their role in viral transmission.