Mechanisms underlying gut microbiota-host interactions in insects.

Insects are the most diverse group of animals and colonize almost all environments on our planet. This diversity is reflected in the structure and function of the microbial communities inhabiting the insect digestive system. As in mammals, the gut microbiota of insects can have important symbiotic functions, complementing host nutrition, facilitating dietary breakdown or providing protection against pathogens. There is an increasing number of insect models that are experimentally tractable, facilitating mechanistic studies of gut microbiota-host interactions. In this Review, we will summarize recent findings that have advanced our understanding of the molecular mechanisms underlying the symbiosis between insects and their gut microbiota. We will open the article with a general introduction to the insect gut microbiota and then turn towards the discussion of particular mechanisms and molecular processes governing the colonization of the insect gut environment as well as the diverse beneficial roles mediated by the gut microbiota. The Review highlights that, although the gut microbiota of insects is an active field of research with implications for fundamental and applied science, we are still in an early stage of understanding molecular mechanisms. However, the expanding capability to culture microbiomes and to manipulate microbe-host interactions in insects promises new molecular insights from diverse symbioses.

Immune system stimulation by the gut symbiont Frischella perrara in the honey bee (Apis mellifera).

Gut bacteria engage in various symbiotic interactions with their host and impact gut immunity and homeostasis in different ways. In honey bees, the gut microbiota is composed of a relatively simple, but highly specialized bacterial community. One of its members, the gammaproteobacterium Frischella perrara induces the so-called scab phenotype, a dark-coloured band that develops on the epithelial surface of the pylorus. To understand the underlying host response, we analysed transcriptome changes in the pylorus in response to bacterial colonization. We find that, in contrast to the gut bacterium Snodgrassella alvi, F. perrara causes strong activation of the host immune system. Besides pattern recognition receptors, antimicrobial peptides and transporter genes, the melanization cascade was upregulated by F. perrara, suggesting that the scab phenotype corresponds to a melanization response of the host. In addition, transcriptome analysis of hive bees with and without the scab phenotype showed that F. perrara also stimulates the immune system under in-hive conditions in the presence of other gut bacterial species. Collectively, our study demonstrates that the presence of F. perrara influences gut immunity and homeostasis in the pylorus. This may have implications for bee health, because F. perrara prevalence differs between colonies and increased abundance of this bacterium has been shown to correlate with dietary alteration and impaired host development. Our transcriptome analysis sets the groundwork for investigating the interplay of bee gut symbionts with the host immune system.

Fractal analysis of extracellular matrix for observer-independent quantification of intestinal fibrosis in Crohn's disease.

Prevention of intestinal fibrosis remains an unresolved problem in the treatment of Crohn's disease (CD), as specific antifibrotic therapies are not yet available. Appropriate analysis of fibrosis severity is essential for assessing the therapeutic efficacy of potential antifibrotic drugs. The aim of this study was to develop an observer-independent method to quantify intestinal fibrosis in surgical specimens from patients with CD using structural analysis of the extracellular matrix (ECM). We performed fractal analysis in fibrotic and control histological sections of patients with surgery for CD (n = 28). To specifically assess the structure of the collagen matrix, polarized light microscopy was used. A score to quantify collagen fiber alignment and the color of the polarized light was established. Fractal dimension as a measure for the structural complexity correlated significantly with the histological fibrosis score whereas lacunarity as a measure for the compactness of the ECM showed a negative correlation. Polarized light microscopy to visualize the collagen network underlined the structural changes in the ECM network in advanced fibrosis. In conclusion, observer-independent quantification of the structural complexity of the ECM by fractal analysis is a suitable method to quantify the degree of intestinal fibrosis in histological samples from patients with CD.

Integration host factor regulates colonization factors in the bee gut symbiont Frischella perrara.

Bacteria colonize specific niches in the animal gut. However, the genetic basis of these associations is often unclear. The proteobacterium Frischella perrara is a widely distributed gut symbiont of honey bees. It colonizes a specific niche in the hindgut and causes a characteristic melanization response. Genetic determinants required for the establishment of this association, or its relevance for the host, are unknown. Here, we independently isolated three point mutations in genes encoding the DNA-binding protein integration host factor (IHF) in F. perrara. These mutants abolished the production of an aryl polyene metabolite causing the yellow colony morphotype of F. perrara. Inoculation of microbiota-free bees with one of the mutants drastically decreased gut colonization of F. perrara. Using RNAseq, we found that IHF affects the expression of potential colonization factors, including genes for adhesion (type 4 pili), interbacterial competition (type 6 secretion systems), and secondary metabolite production (colibactin and aryl polyene biosynthesis). Gene deletions of these components revealed different colonization defects depending on the presence of other bee gut bacteria. Interestingly, one of the T6SS mutants did not induce the scab phenotype anymore despite colonizing at high levels, suggesting an unexpected role in bacteria-host interaction. IHF is conserved across many bacteria and may also regulate host colonization in other animal symbionts.

Laser Alloying Advantages by Dry Coating Metallic Powder Mixtures with SiOx Nanoparticles.

Up to now, minimizing segregation of free-flowing, microscale metal powder mixtures driven by different mass density is an open challenge. In this work, effects of particle size variation on homogeneity of Al-Cu mixtures, with a density ratio of 3.3, are examined. Dry coating Al particles with 0.3 wt% fumed silica SiOx nanoparticles significantly decreases interparticle attraction. This enlarges the range of free-flowing Al particle sizes to < 20 µm. Powder mixture homogeneity is examined optically in vibrated bulk powder and thinly spread layers. From various powder mixtures, solid samples are built layer by layer with the Additive Manufacturing (3D printing) technology Laser Beam Melting in metal powder bed (LBM). Chemical homogeneity of solids is evaluated via energy-dispersive X-ray spectroscopy, backscattered electron microscopy, metallographic analysis and tensile tests. Persistent homogeneity of Al-Cu powder mixtures and LBM solids is found only with particles < 20 µm dry coated with SiOx nanoparticles. Observed segregation phenomena are explained with a decrease in particle mobility at increasing local concentration and the decreasing effectiveness of mass in smaller particles. The main effects are based on geometry, so they are expected to be transferrable to other nanoparticles, alloying components and powder bed technologies, e.g., binder jetting.

Probiotic Treatment with a Gut Symbiont Leads to Parasite Susceptibility in Honey Bees.

Gut symbionts are critical for host health and as such might be used as probiotics. In a recent study, Schwarz et al. showed that pretreatment of honey bees with a dominant gut bacterium causes dysbiosis and increases pathogen susceptibility, showing that probiotic applications for animal health can have unwanted effects.