Working Group “Violet”. Refused-Derived Fuel from Municipal Solid Waste to reduce landfills (REFREsh DANUBE)

The REFREsh Danube project aims to reduce the landfilled amount of municipal solid waste, which is a source of water and soil pollution, by its use as refuse-derived fuel in the cement industry of Novi Sad (Serbia). The main actors involved are Lafarge Beočin Cement Factory, Municipality of Novi Sad, JPK Čistoća (waste-management company) and experts from interdisciplinary fields. The project addresses societal challenges on climate change, air, water and soil pollution as well as using the municipal solid waste as an alternative energy source in response to H2020 and Europe 2020 Strategy

Identification and Characterization of a Novel Family of Cysteine-Rich Peptides (MgCRP-I) from Mytilus galloprovincialis

We report the identification of a novel gene family (named MgCRP-I) encoding short secreted cysteine-rich peptides in the Mediterranean mussel Mytilus galloprovincialis. These peptides display a highly conserved pre-pro region and a hypervariable mature peptide comprising six invariant cysteine residues arranged in three intramolecular disulfide bridges. Although their cysteine pattern is similar to cysteines-rich neurotoxic peptides of distantly related protostomes such as cone snails and arachnids, the different organization of the disulfide bridges observed in synthetic peptides and phylogenetic analyses revealed MgCRP-I as a novel protein family. Genome- and transcriptome-wide searches for orthologous sequences in other bivalve species indicated the unique presence of this gene family in Mytilus spp. Like many antimicrobial peptides and neurotoxins, MgCRP-I peptides are produced as pre-propeptides, usually have a net positive charge and likely derive from similar evolutionary mechanisms, that is, gene duplication and positive selection within the mature peptide region; however, synthetic MgCRP-I peptides did not display significant toxicity in cultured mammalian cells, insecticidal, antimicrobial, or antifungal activities. The functional role of MgCRP-I peptides in mussel physiology still remains puzzling

Determining Obelia spp. diversity and population dynamics in Thau lagoon (Northwestern Mediterranean Sea, France).

The jellyfish Obelia belongs to the family Campanulariidae (Hydrozoa, Leptomedusae), an important and widely distributed family of hydrozoans. Obelia is a very popular and widespread medusa, however first report on its complete life cycle was only published in the late nineties in Northern Japan. This is probably due to its size, as the medusa umbrella diameter is lower than 1mm. This Hydrozoa presents a bentho-pelagic life cycle with both polyp and medusa stages. In Thau lagoon, Northwestern Mediterranean Sea, populations of Obelia occur each year. Some polyps colonies were identified and sampled in the lagoon on Zostera noltii leaves, indicating that the populations realise their full development cycle within the lagoon. Morphological and genetical (COI) approaches, on both polyps and medusae, have revealed that main populations are composed by O. dichotoma individuals (side branches typically irregular in length; hydroteca bell-shaped, usually not very deep, thin walled, often thrown into fine longitudinal folds; hydrotecal rim with smooth or with shallow cusps \u2013crenate, slightly flared; diaphragm transverse to oblique) but that O. bidentata (lateral branches roughly in right angles pairs are given on both sides; slightly oblique diaphragm; hydrotecal rim with bimucronate cusps) is also present in lower abundances. An in situ pelagic monitoring every two weeks since 2008 have allowed understanding the populations dynamics of the genus and the environmental factors that seemed to be particularly appropriated for promoting optimum growth conditions. Baseline information regarding seasonal cycles and historical abundances were provided from a 8 years monitoring, which has allowed detecting blooms. Indeed, the medusae were found at very low densities during the study period with main abundances under 5 ind.m-3, in accordance to previous reports for Obelia spp. medusae around the world. Nevertheless, 2 exceptional events took place in June 2008 and May 2013 were blooms reached 1232 and 660 ind.m-3 respectively. Such data raise questions regarding the potential importance of this small and understudied hydrozoa in the pelagic community of Thau lagoon. Consequently, it is necessary to improve the available databases documenting medusae blooms, as this will provide baseline information about these little-studied events

Aspetti della biologia ed analisi del ciclo riproduttivo di Gobio benacensis (Pollini, 1816) nel Nordest Italia

Il presente studio si propone di approfondire la biologia ed il ciclo riproduttivo del gobione italiano (Gobio benacensis), specie endemica a corologia cisalpina inclusa nella categoria \u201cEndangered\u201d (EN) della lista rossa IUCN, con l\u2019intento di fornire informazioni utili a redigere corretti piani gestionali volti alla conservazione della specie. Le indagini sono state condotte in un corso d\u2019acqua del Bacino del Fiume Isonzo (Torrente Reca, Nordest Italia) ed hanno riguardato 86 esemplari catturati nell\u2019aprile 2016 al fine di definirne la consistenza e la struttura della popolazione e 77 esemplari, catturati mensilmente e bimestralmente tra aprile 2016 e febbraio 2017, per la definizione del ciclo riproduttivo. Le curve di regressione del peso sulla lunghezza totale per i maschi (W=0,1253TL2,8770) e per le femmine (W=0,1555TL2,7554), confrontate mediante ANCOVA, non hanno denunciato differenze significative. I valori medi \ub1 deviazione standard dell\u2019indice gonadosomatico GSI per i maschi sono compresi tra 0,61\ub10,49 e 1,75\ub10,58, mentre per le femmine tra 1,02\ub10,88 e 14,15\ub12,80. L\u2019indice epatosomatico HSI ha denunciato range compresi tra 0,05\ub10,02 e 2,68\ub11,26 e tra 0,25\ub10,19 e 1,75\ub10,58 rispettivamente per i maschi e per le femmine. Le analisi istologiche di ovari e testicoli, analizzati per la prima volta nella specie, hanno permesso di identificare gli stadi di maturazione, che unitamente all\u2019analisi del GSI indicano che la specie depone le uova a pi\uf9 riprese durante il periodo riproduttivo compreso tra aprile e giugno. Nei maschi \ue8 stata, tuttavia, evidenziata una prolungata attivit\ue0 riproduttiva che si protrae fino al mese di agosto

SARS-CoV-2 RNA Recovery from Air Sampled on Quartz Fiber Filters: A Matter of Sample Preservation?

The airborne route of transmission of SARS-CoV-2 was confirmed by the World Health Organization in April 2021. There is an urge to establish standardized protocols for assessing the concentration of SARS-CoV-2 RNA in air samples to support risk assessment, especially in indoor environments. Debates on the airborne transmission route of SARS-CoV-2 have been complicated because, among the studies testing the presence of the virus in the air, the percentage of positive samples has often been very low. In the present study, we report preliminary results on a study for the evaluation of parameters that can influence SARS-CoV-2 RNA recovery from quartz fiber filters spotted either by standard single-stranded SARS-CoV-2 RNA or by inactivated SARS-CoV-2 virions. The analytes were spiked on filters and underwent an active or passive sampling; then, they were preserved at −80 °C for different numbers of days (0 to 54) before extraction and analysis. We found a mean recovery of 2.43%, except for the sample not preserved (0 days) that showed a recovery of 13.51%. We found a relationship between the number of days and the recovery percentage. The results presented show a possible issue that relates to the quartz matrix and SARS-CoV-2 RNA recovery. The results are in accordance with the already published studies that described similar methods for SARS-CoV-2 RNA field sampling and that reported non-detectable concentrations of RNA. These outcomes could be false negatives due to sample preservation conditions. Thus, until further investigation, we suggest, as possible alternatives, to keep the filters: (i) in a sealed container for preservation at 4 °C; and (ii) in a viral transport medium for preservation at a temperature below 0 °C.This research was funded by University of Trieste Atheneum Fund for scientific research (2021) and IRCCS Burlo Garofolo (RC47/20)

dentification of host-pathogen interacting molecules of Mytilus galloprovincialis (Lamark, 1819) using phage-display technology.

2013/2014Le cellule di mollusco immunocompetenti, in primis gli emociti circolanti, provvedono ad una rapida e robusta risposta difensiva nel confronti dei potenziali patogeni. Una volta che gli emociti vengono attivati dall'interazione tra pattern molecolari (PAMPs) presenti sulla superficie dei patogeni e specifici PRRs (pattern recognition receptors) in grado di riconoscerli, queste cellule innescano reazioni difensive. Nonostante un numero sempre più elevato di molecole in grado di interagire con i PAMPs sia stato caratterizzato in M. galloprovincialis, ad oggi non è mai stato effettuato uno studio di interattomica per l’identificazione su larga scala dei PRRs di mitilo coinvolti nel riconoscimento di specifici patogeni. Lo scopo di questo studio è, dunque, quello di identificare i PRRs delle cellule di mollusco immunocompetenti coinvolti nel riconoscimento dei batteri Vibrio splendidus e V. aestuarianus, Gram-negativi presenti in acque costiere e associati ai casi di mortalità che hanno colpito gli allevamenti di ostriche in tutto il mondo e verso i quali i mitili mostrano, invece, notevole resistenza . Per eseguire questo tipo di analisi è stata utilizzata, in modo innovativo, la tecnica phage-display, che si basa sulla possibilità di far esprimere ad un batteriofago un peptide esogeno in fusione con una delle proteine del capside, in modo che la particella fagica esponga sulla sua stessa superficie il peptide di interesse. Il nuovo approccio utilizzato in questo studio ha permesso lo studio diretto dell’interazione tra i fagi recanti un pool di peptiti espressi da emociti di mitilo e i PAMPs presenti sulla superficie delle cellule batteriche. Mediante successive fasi di selezione e amplificazione delle particelle fagiche in grado di legarsi alla superficie dei batteri, è stato possibile arricchire la frazione di cDNA di mitilo codificante PRRs. Con tecniche di sequenziamento massivo e strumenti bioinformatici è stato poi possibile risalire a tutte le sequenze codificanti i peptidi selezionati. I risultati ottenuti indicano che vi è una notevole differenza tra il numero di PRRs di emociti di mitilo che ha interagito con V. splendidus ripetto a V. aestuarianus. Lo studio si è, quindi, incentrato sui 42 peptidi selezionati contro V. splendidus, presunti PRRs, identificandone alcuni con funzione immunitaria già nota (C-type lectin, FREPs, C1qDC proteina, apextrin-related proteina), alcuni presumibilmente falsi positivi ed altri completamente nuovi che non mostrano similarità con sequenze omologhe annotate e il cui ruolo e funzione andrebbero indagati con futuri studi sperimentali.XXVII Ciclo198

Otter diet and prey selection in a recently recolonized area assessed using microscope analysis and DNA barcoding

Ecologists traditionally investigate the feeding ecology of vertebrates by visual identification of prey in their scats using a microscope. This analysis, however, presents some pitfalls, such as poor prey identification at genus/species level. DNA barcoding is an alternative to the traditional diet analysis, recently applied to investigate the diet of several vertebrates. It allows the identification of cryptic or not-recognizable prey portions basing their identification on the analysis of conserved DNA fragments. Here, we combined microscope analysis with DNA barcoding to investigate the diet of the Eurasian otter (Lutra lutra) in an area recently recolonized by the species, in the surroundings of Tarvisio, NE Italy. We collected 102 spraints and analysed all of them at microscope. A random subsample (N=50) was then analysed through DNA barcoding, via amplification of the V9 region of the SSU 18S-rDNA 18S gene. Furthermore, we assessed seasonal variation in otter diet and whether otters select certain salmonids size classes, to identify potential conflict with anglers. According to the microscope analysis, otters ate primarily fish (salmonids and European bullhead), secondarily amphibians and, more rarely, crayfish, both annually and seasonally. DNA barcoding analysis confirmed the microscope results, attributing some salmonids sequences to the brown trout (Salmo trutta). The lack of potential prey species reference sequences in the GenBank database hampered us in identifying the other taxa found in the spraints at more informative levels. Consumed salmonids and bullheads were of small size. In particular, otters selected salmonids size class of 100 mm, which is much smaller than the minimum catchable size for angling. Overall, our study confirms DNA barcoding potential for otter diet analyses, highlighting the need of additional markers to improve molecular prey identification and suggesting that otters do not constitute a significant threat to anglers in the area

Genetic diversity and structure of Megabalanus azoricus in the Azores: Implications for aquaculture management

Megabalanus azoricus giant barnacles are the most traditional seafood of the Azores archipelago (NE Atlantic). This valuable commercial species has been highly exploited in the past and it is considered one of the key species for the development of aquaculture in the region. Despite the importance for conservation and aquaculture there is still a lack of basic information about M. azoricus genetic diversity and population structure. Here we used seven microsatellites markers to analyse 300 samples collected at six out of nine islands of the Azores archipelago, including also different locations from a single island, to provide information on the scale of genetic diversity and population structure of this species. Parameters like heterozygosity, allelic richness and effective number of alleles indicated a high genetic diversity and variability among islands. Pairwise comparisons and PCoA analysis on FST and Jost's DEST showed significant and evident differentiation among sampling locations. Additionally, AMOVA allocates a small (6.02%) but statistically significant portion of the variance to the among Island level revealing also a weak resolution (1.87%) at finer scale. Additionally Monte Carlo resampling methods indicated the most likely sources of the recruits were the local or adjacent populations. Genetic risks associated with the giant barnacle potential production scheme should be taken into account in a future management plan delimiting, as precautionary measure, this culture at a single island or at groups of islands here identified. Moreover a monitoring strategy should be implemented with the aim to evaluate possible changes in genetic parameters of native populations

Drought-induced dieback of Pinus nigra: a tale of hydraulic failure and carbon starvation

10siOngoing climate change is apparently increasing tree mortality rates, and understanding mechanisms of drought-induced tree decline can improve mortality projections. Differential drought impact on conspecific individuals within a population has been reported, but no clear mechanistic explanation for this pattern has emerged. Following a severe drought (summer 2012), we monitored over a 3-year period healthy (H) and declining (D) Pinus nigra trees co-occurring in a karstic woodland to highlight eventual individual-specific physiological differences underlying differential canopy dieback. We investigated differences in water and carbon metabolism, and xylem anatomy as a function of crown health status, as well as eventual genotypic basis of contrasting drought responses. H and D trees exploited the same water pools and relied on similar hydraulic strategies to cope with drought stress. Genetic analyses did not highlight differences between groups in terms of geographical provenance. Hydraulic and anatomical analyses showed conflicting results. The hydraulic tracheid diameter and theoretical hydraulic conductivity were similar, but D trees were characterized by lower water transport efficiency, greater vulnerability to xylem conduit implosion and reduced carbohydrate stores. Our results suggest that extreme drought events can have different impacts on conspecific individuals, with differential vulnerability to xylem embolism likely playing a major role in setting the fate of trees under climate change.openopenSavi, Tadeja; Casolo, Valentino; Dal Borgo, Anna; Rosner, Sabine; Torboli, Valentina; Stenni, Barbara; Bertoncin, Paolo; Martellos, Stefano; Pallavicini, Alberto; Nardini, AndreaSavi, Tadeja; Casolo, Valentino; Dal Borgo, Anna; Rosner, Sabine; Torboli, Valentina; Stenni, Barbara; Bertoncin, Paolo; Martellos, Stefano; Pallavicini, Alberto; Nardini, Andre

The conservation of Apis mellifera (Linnaeus, 1758) in the Karst and Istria

Genetic analyses conducted on bee samples from Istrian Karst have revealed the presence of a high degree of genetic reshuffling of local populations due to repeated imports of non-native bees for commercial purposes. Nevertheless, genetic traces of two presumably native ecotypes are still present. The first is a hybrid between the yellow Italian bee and the grey Carniolan bee and was found mainly in the area of the Karst above Trieste, the natural boundary between the two subspecies. The second, more abundant in Istria, could be the Istrian-Dalmatian ecotype, which is described in the literature as a coastal form of the Carniolan bee adapted to the warmer drier climate of the coastal Karst. These two ecotypes are morphologically very similar, but distinct from a genetic perspective. The identification and characterization of these local strains is a first step to implementing targeted programmes for the conservation and the restoration of their rearing. It then becomes of primary importance to create regulations that prohibit the introduction of non-native bees in the area to curb the risk of extinction of local varieties