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- Insect biotechnology (2)
- Keratin (2)
- Tineola bisselliella (2)
- Annual rings (1)
- Archäologischer Nassholzfund (1)
- Art restauration (1)
- Beneficial microbes (1)
- Cold to fast neutron testing (1)
- Dietary adaptation (1)
- Gene expression (1)
Organisationseinheit der BAM
The clothes moth Tineola bisselliella is one of a few insects that can digest keratin, leading to the destruction of clothing, textiles and artwork. The mechanism of keratin digestion is not yet fully understood, partly reflecting the lack of publicly available genomic and transcriptomic data. Here we present a high-quality gut transcriptome of T. bisselliella generated from larvae reared on keratin-rich and keratin-free diets. The overall transcriptome consists of 428,221 contigs that were functionally annotated and screened for candidate enzymes involved in keratin utilization. As a mechanism for keratin digestion, we identified cysteine synthases, cystathionine β-synthases and cystathionine γ-lyases. These enzymes release hydrogen sulfite, which may reduce the disulfide bonds in keratin. The dataset also included 27 differentially expressed contigs with trypsin domains, among which 20 were associated with keratin feeding. Finally, we identified seven collagenases that were upregulated on the keratin-rich diet. In addition to this enzymatic repertoire potentially involved in breaking down keratin, our analysis of poly(A)-enriched and poly(A)-depleted transcripts suggested that T. bisselliella larvae possess an unstable intestinal microbiome that may nevertheless contribute to keratin digestion
While the absorption of X-rays and gamma radiation is determined by the Z-number of the elements a specimen is composed of, it is the hydrogen making an effective contrast with neutron imaging. As a consequence, interrogating with neutrons presents a suitable tool to study the distribution of organic consolidants in materials such as wood as encountered in impregnated wooden artworks.
Four different examples of objects are presented here to demonstrate the potential of neutron CT: 1) small wooden pieces of ship wrecks (< 2 cm thickness) interrogated with cold neutrons (0.5 meV at the ANTARES facility of the FRM II in Garching) to demonstrate the potential and the limitation of using low energy neutrons, 2) a wooden statue soaked with carbolineum (fission neutrons 1.8 MeV at the NECTAR facility of the FRM II), 3) a smaller wooden figure of a skull heavily soaked with carbolineum so it was too tight for the fission neutrons used before with accelerator neutrons (broad range about 5.5 MeV at the PTB in Braunschweig) and 4) pieces of charred wood to study the impregnation with a consolidant (NECTAR, FRM II). With the exception of the last example, all results have been combined with X-ray tomography (BAM 8.3 in Berlin). In the case of the charred wood specimens (example 4) the density histograms of the neutron tomography results were compared with those obtained from untreated references. The observed gain in specific density of the soaked specimens corresponded with an increase of specific weight. All results obtained so far showed distinct distribution patterns attributable to structural peculiarities or organic consolidants providing valuable support for subsequent restoration works.
The effective high neutron scattering absorption coefficient of hydrogen (48.5 cm²/g) due to the scattering allows neutrons to reveal hydrocarbon structures with more contrast than X-rays, but at the same time limits the sample size and thickness that can be investigated. Many planar shaped objects, particularly wood samples, are sufficiently thin to allow thermal neutrons to transmit through the sample in a direction perpendicular to the planar face but not in a parallel direction, due to increased thickness. Often, this is an obstacle that prevents some tomographic reconstruction algorithms from obtaining desired results because of inadequate information or presence of distracting artifacts due to missing projections. This can be true for samples such as the distribution of glue in glulam (boards of wooden layers glued together), or the course of partially visible annual rings in trees where the features of interest are parallel to the planar surface of the sample. However, it should be possible to study these features by rotating the specimen within a limited angular range. In principle, this approach has been shown previously in a study with fast neutrons [2]. A study of this kind was performed at the Antares facility of FRM II in Garching with a 2.6×107/cm² s thermal neutron beam. The limit of penetration was determined for a wooden step wedge carved from a 2 cm×4 cm block of wood in comparison to other materials such as heavy metals and Lucite as specimens rich in hydrogen. The depth of the steps was 1 cm, the height 0.5 cm. The annual ring structures were clearly detectable up to 2 cm thickness. Wooden specimens, i.e. shivers, from a sunken old ship have been subjected to tomography. Not visible from the outside, clear radial structures have been found that are typical for certain kinds of wood. This insight was impaired in a case where the specimen had been soaked with ethylene glycol. In another large sample study, a planar board made of glulam has been studied to show the glued layers. This study shows not only the limits of penetration in wood but also demonstrates access to structures perpendicular to the surface in larger planar objects by tomography with fast neutrons, even with incomplete sets of projection data that covers an angular range of only 90° or even 60°.
The evolutionary success of insects is promoted by their association with beneficial microbes that enable the utilization of unusual diets. The synanthropic clothing moth Tineola bisselliella provides an intriguing example of this phenomenon. The caterpillars of this species have adapted to feed on keratin-rich diets such as feathers and wool, which cannot be digested by most other animals and are resistant to common digestive enzymes. Inspired by the hypothesis that this ability may be conferred by symbiotic microbes, we utilized a simple assay to detect keratinase activity and a method to screen gut bacteria for candidate enzymes, which were isolated from feather-fed larvae. The isolation of DNA from keratin-degrading bacterial strains followed by de novo genome sequencing resulted in the identification of a novel bacterial strain related to Bacillus sp. FDAARGOS_235. Genome Annotation identified 20 genes with keratinase domains. Proteomic analysis of the culture supernatant from this gut bacterium grown in non-nutrient buffer supplemented with feathers revealed several candidate enzymes potentially responsible for keratin degradation, including a thiol-disulfide oxidoreductase and multiple proteases. Our results suggest that the unusual diet of T. bisselliella larvae promotes their association with keratinolytic microorganisms and that the ability of larvae to feed on keratin can at least partially be attributed to bacteria that produce a cocktail of keratin-degrading enzymes.