Environmental DNA assessment of airborne plant and fungal seasonal diversity

Environmental DNA (eDNA) metabarcoding and metagenomics analyses can improve taxonomic resolution in biodiversity studies. Only recently, these techniques have been applied in aerobiology, to target bacteria, fungi and plants in airborne samples. Here, we present a nine-month aerobiological study applying eDNA metabarcoding in which we analyzed simultaneously airborne diversity and variation of fungi and plants across five locations in North and Central Italy. We correlated species composition with the ecological characteristics of the sites and the seasons. The most abundant taxa among all sites and seasons were the fungal genera Cladosporium, Alternaria, and Epicoccum and the plant genera Brassica, Corylus, Cupressus and Linum, the latter being much more variable among sites. PERMANOVA and indicator species analyses showed that the plant diversity from air samples is significantly correlated with seasons, while that of fungi varied according to the interaction between seasons and sites. The results consolidate the performance of a new eDNA metabarcoding pipeline for the simultaneous amplification and analysis of airborne plant and fungal particles. They also highlight the promising complementarity of this approach with more traditional biomonitoring frameworks and routine reports of air quality provided by environmental agencies

Two benthic diatoms, nanofrustulum shiloi and striatella unipunctata, encapsulated in alginate beads, influence the reproductive efficiency of paracentrotus lividus by modulating the gene expression

Physiological effects of algal metabolites is a key step for the isolation of interesting bioactive compounds. Invertebrate grazers may be fed on live diatoms or dried, pelletized, and added to compound feeds. Any method may reveal some shortcomings, due to the leaking of wound-activated compounds in the water prior to ingestion. For this reason, encapsulation may represent an important step of bioassay-guided fractionation, because it may assure timely preservation of the active compounds. Here we test the effects of the inclusion in alginate (biocompatible and non-toxic delivery system) matrices to produce beads containing two benthic diatoms for sea urchin Paracentrotus lividus feeding. In particular, we compared the effects of a diatom whose influence on P. lividus was known (Nanofrustulum shiloi) and those of a diatom suspected to be harmful to marine invertebrates, because it is often present in blooms (Striatella unipunctata). Dried N. shiloi and S. unipunctata were offered for one month after encapsulation in alginate hydrogel beads and the larvae produced by sea urchins were checked for viability and malformations. The results indicated that N. shiloi, already known for its toxigenic effects on sea urchin larvae, fully conserved its activity after inclusion in alginate beads. On the whole, benthic diatoms affected the embryogenesis of P. lividus, altering the expression of several genes involved in stress response, development, skeletogenesis and detoxification processes. Interactomic analysis suggested that both diatoms activated a similar stress response pathway, through the up-regulation of hsp60, hsp70, NF-κB, 14-3-3 ε and MDR1 genes. This research also demonstrates that the inclusion in alginate beads may represent a feasible technique to isolate diatom-derived bioactive compounds

Immunodetection of IgM, IgT and pIgR in mucosal tissues of Antarctic teleost

We have previously investigated the immune response at hepato-biliary level in the Antarctic teleost Trematomus bermacchii, a species belonging to the Perciform suborder Notothenoidei, the most abundant component of the fish fauna living in the Antarctic ocean. By that time only the IgM isotype was known and well characterized at molecular and biochemical levels in Antarctic fish. Over the past few years we have cloned and sequenced genes encoding other two key molecules of the mucosal immune system, IgT and polymeric Ig receptor (pIgR) of T. bernacchii. The present study aimed at investigating the localization in mucosal tissues of IgM, IgT and pIgR in an attempt to clarify the protein occurrence and transepithelial transport. Biochemical and immunohistochemical data provided convergent data about specific mechanisms operating apical release of IgT in exocrine way, as well as depicting peculiar (maybe ancestral) features compared with well- known mechanisms described for polymeric Igs transport in mammalian tissues

Integrated eDNA metabarcoding and morphological analyses assess spatio-temporal patterns of airborne fungal spores

Fungi represent relevant allergens and plant pathogens that can disperse on long ranges, potentially producing severe consequences on public health and agriculture. Up to 11% of the bioaerosol particles are fungal spores and mycelium fragments. Estimation of fungal species diversity in time and space is decisive but may be biased by abiotic conditions and sampling methods. Traditional morphological analyses of fungal spores have been widely applied in aerobiology in the past, while recently eDNA metabarcoding can complement these studies. Here, we used both morphological analysis (spore count and taxon identification) and high-throughput sequencing to disentangle spatio-temporal variation of fungi across Northern and Central Italy and to evaluate the detection efficiency of the two approaches. Our results showed that eDNA metabarcoding detects about three times more genera and has a higher detection efficiency than the morphological analyses. However, the efficiency is high in both spore count and eDNA metabarcoding methods when the most abundant or the rarest genera are considered but it can substantially vary between the two approaches when moderately abundant genera are analyzed. Furthermore, morphological spore determination resulted in higher variance explained by PERMANOVA analysis with respect to eDNA metabarcoding (26% and 13%, respectively), which leads to a better spatio-temporal characterization of the fungal genera. As both morphological analyses and eDNA metabarcoding methods capture significant interactions between seasons and sites, they could be preferably used as complementing approaches to reliably study airborne fungal diversity and variation

Thermoluminescent dosimeters (TLDs-100) calibration for dose verification in photon and proton radiation therapy

Thermoluminescent dosimeters (TLDs) are practical, accurate, and precise tools for point dosimetry in medical physics applications. TLDs are nowadays extensively used to measure dose in conformal radiation therapy in order to guarantee the safety of the treatment. Several national and international organizations recommend checking the effective dose delivered to an individual patient by means of in vivo dosimetry. Modern radiotherapy techniques employing both photon and ion beams exhibit excellent target conformation throught high steep-dose gradients between tumour and adjacent organs and tissues. In this context, catching potential dose errors and uncertainties in treatment delivering is the first step to ensure the optimization of the treatment plan. This study shows the results of the characterization of TLDs-100 at two Italian facilities devoted to advanced radiation treatments with photon and proton therapy. The individual sensitivity factor was determined, and the calibration curves were carried out in the dose range 0–20 Gy. By the analysis of the dose response curves, the linear region was identified under the dose level of 10 Gy. Characterization of the TLDs-100 has enabled their use for in vivo dosimetry especially in the dose range corresponding to the linear region of the dose response curves