Microplastics in deep water : a combined GC-MS e FT-IR approach

Microplastics are water-insoluble, solid polymer particles that are 645\u202fmm in size. A lower size boundary has not been defined, but particles below 1\u202f\u3bcm are usually referred to as nanoplastics rather than microplastics. The ubiquity of microplastics of all sizes in surface water, groundwater and wastewater has raised the question if pollution of drinking water occurs and to date, few studies indeed described the presence of this new contaminant in tap and bottled water. Toxicity it is not yet known, they might cause local inflammations in the gut, but a transport into organs might occur. As a final problem in this field, microplastics in the environment are difficult to sample, to identify and standardized methods do not exist. For this reason, we have been involved in a research project finalized to the definition of a protocol strategy for sampling and analysis of microplastics in drinking waters, coming from deep water wells, in the networks of three large cities in the Northern part of Italy. The project is on going and presently two groups of deep water samples have been collected in the city of Milano and Brescia, before and after the treatment stages used for the urban network. Great attention has been paid to sampling and extraction steps as microplastic contamination, in this kind of samples, is expected to be very low and contamination during sampling might occur through air or materials. Analytical methods have been defined for the detection of main microplastic contamination, like PE/PP, PS, PA, PVC e PET residues > 1 \ub5m. Samples have been analysed first by a non-destructive approach using a Spotlight 200i microscope equipped with a \ub5ATR probe and coupled to an FT-IR spectrophotometer, followed by a mass spectrometric characterization of the polymers by a solid phase microextration (SPME) GC-MS of thermal decomposition products (TED) of microplastic residues. Results will be presented and critically discussed

Identification of differentially expressed small RNAs and prediction of target genes in Italian Large White pigs with divergent backfat deposition

The identification of the molecular mechanisms regulating pathways associated with the potential for fat deposition in pigs can lead to the detection of key genes and markers for the genetic improvement of fat traits. Interactions of microRNAs (miRNAs) with target RNAs regulate gene expression and modulate pathway activation in cells and tissues. In pigs, miRNA discovery is far from saturation, and the knowledge of miRNA expression in backfat tissue and particularly of the impact of miRNA variations is still fragmentary. Using RNA-seq, we characterized the small RNA (sRNA) expression profiles in Italian Large White pig backfat tissue. Comparing two groups of pigs divergent for backfat deposition, we detected 31 significant differentially expressed (DE) sRNAs: 14 up-regulated (including ssc-miR-132, ssc-miR-146b, ssc-miR-221-5p, ssc-miR-365-5p and the moRNA ssc-moR-21-5p) and 17 down-regulated (including ssc-miR-136, ssc-miR-195, ssc-miR-199a-5p and ssc-miR-335). To understand the biological impact of the observed miRNA expression variations, we used the expression correlation of DE miRNA target transcripts expressed in the same samples to define a regulatory network of 193 interactions between DE miRNAs and 40 DE target transcripts showing opposite expression profiles and being involved in specific pathways. Several miRNAs and mRNAs in the network were found to be expressed from backfat-related pig QTL. These results are informative for the complex mechanisms influencing fat traits, shed light on a new aspect of the genetic regulation of fat deposition in pigs and facilitate the prospective implementation of innovative strategies of pig genetic improvement based on genomic markers

The presence of the brown bear (Ursus arctos, L.) in the north eastern Alps (Friuli Venezia Giulia region) in relation to the Alpine metapopulation

The North Eastern Alps (Friuli Venezia Giulia Region) represent a peripheral area of presence of the Dinaric population (for the North Dinaric population, Slovenia, 400-450 individuals estimated) of the Brown Bear; in this area in the last years some individuals has started to come from Trentino population (40-45 individuals estimated) and to use this areas. In previous 114 researches this area has been considered one of the most suitable areas, in the alps, for supporting the alpine brown bear population and for enhancing the creation of the alpine metapopulation. From 2004 to the 2013 in mountains areas of the Friuli Venezia Giulia has been studied (2500 km2) the presence of the Brown Bear with different techniques: the genetic sampling (by opportunistic and systematic methods), photo interpretation and behavioral analysis and radiotelemetry. The genetic samplings have been obtained by the hair traps (50 permanent hair traps plus 80 temporary and moving hair traps) and by opportunistic techniques. By the genetic analysis it has detected the presence, from 2004, of 21 different genotypes (4 from Trentino and 17 from the Dinaric population, 3 of them studied by telemetry), all of them were males. The number of genotypes detected in different year has showed a cyclic pattern, with a increment in the last year, due to dispersing individuals from Dinaric source population; in different years, from 3 to 8 (2013), different genotypes have been detected, with a evident turnover. The median time of presence (years of sampling), for each genotypes, was 2 years, and only 6 genotypes have been sampled for 3 or more years (1 from Trentino); 10 genotypes have been sampled in only one year. Despite to the increment of the Trentino population and the high density of the Dinaric populations, in the north eastern alps the process of immigration and colonization appears to be slow and intermittent and seems correlated more to the demographic aspects (i.e localization and philopatry of the females, now present only in the Alpine and Dinaric areas of Slovenia and on the right side of Adige river, in Trentino), management of the Dinaric population, dispersal behavior and presence of not permeable infrastructure (high way Ljubljana-Trieste) rather than the ecological values of the areas

[use Of Alcoholic Beverages By Children: A Not Yet Investigated Phenomenon].

The essay discourses on the ethanol prescription to children, their dependence and the scientific data related to the social abuse. The toxic effects, social and economical perspectives are well exemplified on the follow-up of a child of three years old in a university hospital during eighteen months. At the end, the prevalence of such practice is highlighted as essential for preventive approach on public health.7052-

Nero Siciliano pig: analysis of coat colour affecting genes and perspectives for breed traceability

Nero Siciliano is an autochthonous pig breed reared in the internal areas of Sicily region mainly in the Nebrodi mountains. The animals are usually completely black with a dorsal stripe but a few present white portions mainly in the face or in the fore legs. According to the increased requests of the consumers for local and typical products, meat and cured products of Nero Siciliano pigs are sold at a higher price compared to other pig products. Thus there is the need to guarantee both consumers and the whole Nero Siciliano production chain from possible frauds. The identification and/or use of DNA markers that may be breed specific could make it possible to establish breed traceability and authenticity systems for the products obtained with this local pig breed. Mutations in coat colour genes have been already described and utilized for porcine breed traceability. In this trial we analysed mutations identified in two coat colour affecting genes, the melanocortin 1 receptor (MC1R) and the v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene (KIT), with the aim to characterize the Nero Siciliano pig at these loci and provide useful information to establish authenticity systems for the meat products. Fragment analysis of PCR products and PCRRFLP methods were used to identify the polymorphic sites that can distinguish known alleles at these two loci in 104 Nero Siciliano pigs. Four alleles were identified at the MC1R locus: the two dominant black alleles (ED2, frequency of 0.673; ED1, 0.187), allele EP (0.106) and the recessive e allele (0.034). The results showed that different alleles were observed at this locus, polymorphisms at the MC1R gene cannot be used for product traceability and authentication of this breed. As regards the KIT locus, all the animals were negative for the splice site mutation of exon/intron 17. Thus, meat of Nero Siciliano pigs can be distinguished from meat of white pigs that are positive for this polymorphic site. Moreover, at this locus only 4 pigs showed the 3'-5' duplication breakpoint suggesting that they carried the Ip allele. Studies are in progress to evaluate the effect of this allele on coat colour phenotypes in Nero Siciliano pig

Analysis of melanocortin 1 receptor (MC1R) gene polymorphisms in some cattle breeds: their usefulness and application for breed traceability and authentication of Parmigiano Reggiano cheese

l legame tra un prodotto di origine animale e la razza da cui questo \ue8 originato rappresenta un aspetto importante per la valorizzazione di alcune produzioni. Il maggior prezzo che questi prodotti spuntano sul mercato fa emergere l\u2019esigenza di poter autenticare o tracciare i prodotti mono-razza per smascherare e scoraggiare possibili frodi. A questo scopo sono stati proposti sistemi di analisi del DNA, alcuni dei quali utilizzano marcatori in geni che determinano il colore del mantello, che \ue8 uno dei principali caratteri che differenziano tra di loro le razze. Diverse mutazioni nel gene melanocortin 1 receptor (MC1R) sono gi\ue0 state associate a particolari effetti sul colore del mantello nella specie bovina. In questa ricerca abbiamo studiato la presenza dei principali alleli al locus MC1R, per valutare la possibilit\ue0 di utilizzare questo gene per l\u2019autenticazione e la tracciabilit\ue0 di razza dei prodotti lattiero-caseari. Le mutazioni che permettono di distinguere questi alleli sono state analizzate utilizzando protocolli di PCR-RFLP e PCR-APLP su un totale di 1360 animali appartenenti a 18 razze bovine. Per ognuna delle seguenti razze, Frisona Italiana, Bruna Italiana, Pezzata Rossa Italiana, Jersey, Rendena, Reggiana e Modenese, \ue8 stato possibile analizzare pi\uf9 di 70 animali. L\u2019allele Ed \ue8 stato identificato nella razza Frisona Italiana con una frequenza dello 0,886. L\u2019allele E (nomenclatura che include tutti gli alleli tranne che e, Ed e E1) \ue8 stato identificato con alta frequenza nella Bruna Italiana (0,591), Rendena (0,738), Jersey (0,955) e Modenese (0,961) e con bassa frequenza nella Pezzata Rossa Italiana (0,029). Inoltre, questo allele \ue8 stato osservato nella Rossa Svedese, Rossa Danese, Grigio Alpina, Piemontese, Romagnola, Marchigiana e Chianina. In alcune di queste razze (Bruna Italiana, Rendena, Grigio Alpina, Piemontese, Rossa Svedese e Rossa Danese) \ue8 stato identificato anche l\u2019allele E1. L\u2019allele e \ue8 risultato fissato nella razza Reggiana e quasi fissato nella razza Pezzata Rossa Italiana. Inoltre, con bassa frequenza, \ue8 stato identificato in tutte le altre razze analizzate, tranne che nella Marchigiana. Le differenze osservate tra razze esaminate indicano che, almeno in alcuni casi, \ue8 possibile utilizzare i polimorfismi del gene MC1R per escludere o confermare l\u2019impiego di latte di una determinata razza nella produzione di un prodotto lattiero-caseario. Il caso pi\uf9 interessante \ue8 quello del formaggio Parmigiano Reggiano prodotto con l\u2019uso esclusivo di latte di bovine di razza Reggiana. Infatti, essendo presente in questa razza soltanto l\u2019allele e il rilievo analitico di qualsiasi altro allele nel DNA estratto dal formaggio rivela l\u2019uso di latte proveniente da altre razze. La messa a punto di un metodo PCR-RFLP per l\u2019analisi del DNA estratto da prodotti lattiero caseari, incluso il Parmigiano Reggiano di oltre 24 mesi di stagionatura, rappresenta uno strumento importante per la difesa di questo prodotto mono-razza da eventuali frodi. I risultati ottenuti su 10 forme di formaggio prodotto esclusivamente con latte di bovine di razza Reggiana e su 15 forme di Parmigiano Reggiano commerciale ottenuto senza restrizione della razza di origine del latte hanno mostrato la validit\ue0 del metodo del quale \ue8 stata valutata anche la sensibilit\ue0n cattle, the MC1R gene has been the subject of several studies with the aim to elucidate the biology of coat colour. Then, polymorphisms of this gene have been proposed as tools for breed identification and animal products authentication. As a first step to identify breed specific DNA markers that can be used for the traceability of mono-breed dairy cattle products we investigated, using PCR-RFLP and PCR-APLP protocols, the presence and distribution of some alleles at the MC1R locus in 18 cattle breeds for a total of 1360 animals. For each of seven breeds (Italian Holstein, Italian Brown, Italian Simmental, Rendena, Jersey, Reggiana and Modenese) a large number of animals (>70) was genotyped so the obtained results can be considered with more confidence. Allele Ed was identified only in black pied cattle (Italian Holstein and Black Pied Valdostana). Allele E (this nomenclature includes all alleles except Ed, E1 and e) was observed in Italian Brown, Rendena, Jersey, Modenese, Italian Simmental, Grigio Alpina, Piedmontese, Chianina, Romagnola, Marchigiana, Swedish Red and White and Danish Red. Allele E1 was identified in Italian Brown, Rendena, Grigio Alpina, Piedmontese, Swedish Red and White and Danish Red. The recessive allele e, known to cause red coat colour, was fixed in Reggiana and almost fixed in Italian Simmental. This allele was observed also in Italian Holstein, Italian Brown, Rendena, Jersey and Modenese albeit with low frequency. Moreover, this allele was detected in Valdostana, Pezzata Rossa d\u2019Oropa, Piedmontese, Romagnola, Swedish Red and White, Danish Red, Charoleis and Salers. In the case of the Reggiana breed, which is fixed for allele e, the MC1R locus is highly informative with respect to breeds that carry other alleles or in which allele e is at very low frequency. In theory, using the MC1R locus it is possible to identify the presence of milk from some other breeds in Parmigiano Reggiano cheese labelled as exclusively from the Reggiana breed. This possibility was practically tested by setting up protocols to extract and analyse polymorphisms of the MC1R locus in several dairy products, including Parmigiano Reggiano cheese cured for 30 months. The lower detection limit was estimated to be 5% of non expected DNA. This test can represent a first deterrent against fraud and an important tool for the valorisation and authentication of Parmigiano Reggiano cheese obtained from only Reggiana milk

Design and construction of a new metering hot box for the in situ hygrothermal measurement in dynamic conditions of historic masonries

The main purpose of the HeLLo project is to contribute to data available on the literature on the real hygrothermal behavior of historic walls and the suitability of insulation technologies. Furthermore, it also aims at minimizing the energy simulation errors at the design phase and at improving their conservation features. In this framework, one of the preliminary activities of the study is the creation of a real in situ hot box to measure and analyze different insulation technologies applied to a real historic wall, to quantify the hygrothermal performance of a masonry building. Inside this box, 'traditional' experiments can be carried out: recording heat flux, surface temperature, and air temperatures, as well as relative humidity values through the use of a new sensing system (composed of thermocouples and temperature/relative humidity combined sensors). Within this paper, the process of development, construction, and validation of this new metering box is exhibited. The new hot box, specifically studied for historic case studies, when compared to other boxes, presents other advantages compared to previous examples, widely exemplified

Changing patterns of conflict between humans, carnivores and crop-raiding prey as large carnivores recolonize human-dominated landscapes

Large carnivores are making remarkable comebacks in Europe, but how this affects human-wildlife conflict remains unclear. Rebounding carnivore populations lead to increasing livestock depredation, which in turn leads to greater economic losses for farmers. However, returning carnivores could also influence the behavior of wild ungulates, which are themselves responsible for major crop damage and associated economic losses. Here, we exploit the natural experiment of a rebounding wolf population in the Italian Apennines to study how this affected both types of human-wildlife conflic. We used large datasets of wolf occurrences (n = 351), livestock depredation events (n = 165), and crop damage events by wild boar (n = 3442) to independently model the determinants of livestock depredation and crop damage distribution in relation to wolf habitat suitability over a ten-year period of increasing wolf numbers. These analyses yielded two major insights. First, livestock depredations were mainly related to insufficient prevention measures (e.g. lacking fencing) rather than landscape context, providing a clear pathway to conflict mitigation. Second, crop damage decreased in areas of higher wolf habitat suitability and became more likely in areas of lower wolf habitat suitability, closer to settlements. This suggests increasing predation pressure forces wild boars to avoid the most suitable wolf habitat, leading to a redistribution of crop damage in the landscape. More generally, our study highlights complex human-wildlife interactions as large carnivores recover in human-dominated landscapes, suggesting that multiple, cooccurring conflicts need to be assessed jointly and adaptively in order to foster coexistence between humans and wildlife

Investigation of SNPs in the ATP1A2, CA3 and DECR1 genes mapped to porcine chromosome 4: analysis in groups of pigs divergent for meat production and quality traits

STUDIO DI TRE GENI (ATP1A2, CA3E DECR1) LOCALIZZATI SUL CROMOSOMA SUINO 4: ANALISIDELLE FREQUENZE ALLELICHE DI SNP IN SUINI ESTREMI PER ALCUNI CARATTERI PRODUTTIVITre geni (ATPase, Na+/K+transporting, \u3b12(+) polypeptide, ATP1A2; carbonic anhydrase III, CA3; 2,4-die-noyl CoA reductase 1, mitochondrial, DECR1), isolati da una libreria a cDNA ottenuta da muscolo scheletri-co di suino, sono stati scelti per questo studio sulla base del ruolo fisiologico della proteina codificata in pro-cessi cellulari e metabolici collegabili in modo diretto o indiretto con alcuni caratteri produttivi. La scelta dei tre geni \ue8 stata anche basata sulla loro localizzazione sul cromosoma 4 dove diversi QTL per caratteri pro-duttivi sono gi\ue0 stati identificati.Nella regione 3\u2019 non tradotta del gene CA3\ue8 stato identificato, mediante analisi SSCP, un polimorfismo bial-lelico. Il sequenziamento dei due alleli ha permesso di identificare una nuova mutazione puntiforme, che \ue8stata successivamente analizzata mediante PCR-RFLP. Per questo gene \ue8 stato effettuato il mappaggiogenetico sul cromosoma 4 mediante l\u2019analisi della mutazione nei campioni delle famiglie di riferimento delprogetto europeo di mappaggio del genoma suino (PiGMaP). Utilizzando il polimorfismo PCR-RFLP del geneATP1A2, gi\ue0 decritto in un precedente lavoro come SSCP, sono state tipizzate diverse famiglie di riferimen-to PiGMaP ed \ue8 stato confermato il mappaggio genetico di ATP1A2. Utilizzando queste informazioni e quel-le gi\ue0 disponibili per DECR1, per la prima volta, \ue8 stata ottenuta una mappa di linkage del cromosoma 4che comprende tutti e tre i geni analizzati. Il mappaggio genetico dei tre geni \ue8 stato anche confermatomediante tipizzazione di un pannello di ibridi di cellule irradiate (IMpRH 7000 rad).Le frequenze alleliche delle mutazioni identificate nei tre loci sono state studiate in 11 razze suine (LargeWhite Italiana, Landrace Italiana, Duroc Italiana, Landrace Belga, Hampshire, Pi\ue9train, Meishan, CintaSenese, Casertana, Calabrese and Nero Siciliano) per un totale di 272 animali. Inoltre, come approccio ini-ziale per poi scegliere i geni da analizzare in futuri studi di associazione, abbiamo confrontato le frequenzealleliche di mutazioni puntiformi per questi tre loci in gruppi di suini di razza Large White Italiana e DurocItaliana, analizzando animali con valori degli indici genetici estremi e divergenti per alcuni caratteri produt-tivi (accrescimento, spessore lardo dorsale, tagli magri e grasso intermuscolare visibile). Per il gene CA3 \ue8stata osservata una differenza nella distribuzione delle frequenze alleliche (P< 0,05) per i due caratteri taglimagri (nella razza Large White Italiana) e grasso intermusculare visibile (nella razza Duroc Italiana). Per ilgene DECR1, differenze significative sono state osservate per il carattere grasso intermuscolare visibile. Ilgene ATP1A2, che mappa vicino al locus FAT1, non ha presentato differenze statisticamente significativenelle frequenze tra i gruppi estremi per i caratteri oggetto di studio. Analizzando il livello di linkage disequi-librium(LD) tra i tre loci, \ue8 stato evidenziato un elevato livello di LD (D\u2019= 0,967; P< 0,0001) tra i geni CA3e DECR1, solo nella popolazione Duroc Italiana. Questi primi risultati pongono le basi per ulteriori studi perverificare se i geni CA3e DECR1sono associati con i caratteri oggetto di selezione nel suino pesante.Three genes (ATPase, Na+/K+ transporting, \u3b1 2(+) polypeptide, ATP1A2; carbonic anhydrase III, CA3; 2,4-dienoyl CoA reductase 1, mitochondrial, DECR1), isolated from a porcine skeletal muscle cDNA library and mapped on porcine chromosome 4 (SSC4), were investigated. A new single nucleotide polymorphism (SNP) was identified in the 3\u2019-untranslated region of the CA3 gene and used to genetically map this locus on SSC4 together with the ATP1A2 and DECR1 loci for which SNPs were already reported. Allele frequencies of the three loci were reported for 11 pig breeds (Italian Large White, Italian Landrace, Italian Duroc, Belgian Landrace, Hampshire, Pi\uf9train, Meishan, Cinta Senese, Casertana, Calabrese and Nero Siciliano). Radiation hybrid mapping of these genes confirmed the linkage mapping results as well as mapping information reported by other authors. Then, the SNPs identified in the ATP1A2, CA3 and DECR1 genes were genotyped in Italian Large White and Italian Duroc animal groups with extreme and divergent estimated breeding value for several production traits. For CA3 significant differences in allele frequencies (P< 0.05) were observed between the extreme groups of pigs for the lean cuts (Italian Large White) and visible intermuscular fat (Italian Duroc) traits. For DECR1, a significant difference in allele frequencies was observed only for the visible intermuscular fat trait. ATP1A2, which maps close to the FAT1 locus, did not show any significant difference. A very high linkage disequilibrium (D\u2019= 0.967; P< 0.0001) was identified between CA3 and DECR1 in the Italian Duroc population. Further investigations are needed to evaluate the effect of CA3 and DECR1 on the considered trait