PCR-RFLP approaches to easily identify Pleuronectes platessa from other flatfishes: a rapid and efficient tool to control label information

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Sensor-embedded face masks for detection of volatiles in breath: a proof of concept study

The correlation between breath volatilome and health is prompting a growing interest in the development of sensors optimized for breath analysis. On the other hand, the outbreak of COVID-19 evidenced that breath is a vehicle of infection; thus, the introduction of low-cost and disposable devices is becoming urgent for a clinical implementation of breath analysis. In this paper, a proof of concept about the functionalization of face masks is provided. Porphyrin-based sensors are among the most performant devices for breath analysis, but since porphyrins are scarcely conductive, they make use of costly and bulky mass or optical transducers. To overcome this drawback, we introduce here a hybrid material made of conducting polymer and porphyrins. The resulting material can be easily deposited on the internal surface of standard FFP face masks producing resistive sensors that retain the chemical sensitivity of porphyrins implementing their combinatorial selectivity for the identification of volatile compounds and the classification of complex samples. The sensitivity of sensors has been tested with respect to a set of seven volatile compounds representative of diverse chemical families. Sensors react to all compounds but with a different sensitivity pattern. Functionalized face masks have been tested in a proof-of-concept test aimed at identifying changes of breath due to the ingestion of beverages (coffee and wine) and solid food (banana- and mint-flavored candies). Results indicate that sensors can detect volatile compounds against the background of normal breath VOCs, suggesting the possibility to embed sensors in face masks for extensive breath analysis

SARS-CoV-2 has been circulating in northern Italy since december 2019: evidence from environmental monitoring

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is responsible for the coronavirus disease COVID-19, a public health emergency worldwide, and Italy is among the most severely affected countries. The first autochthonous Italian case of COVID-19 was documented on February 21, 2020. We investigated the possibility that SARS-CoV-2 emerged in Italy earlier than that date, by analysing 40 composite influent wastewater samples collected - in the framework of other wastewater-based epidemiology projects - between October 2019 and February 2020 from five wastewater treatment plants (WWTPs) in three cities and regions in northern Italy (Milan/Lombardy, Turin/Piedmont and Bologna/Emilia Romagna). Twenty-four additional samples collected in the same WWTPs between September 2018 and June 2019 (i.e. long before the onset of the epidemic) were included as ‘blank’ samples. Viral concentration was performed according to the standard World Health Organization procedure for poliovirus sewage surveillance, with modifications. Molecular analysis was undertaken with both nested RT-PCR and real-rime RT-PCR assays. A total of 15 positive samples were confirmed by both methods. The earliest dates back to 18 December 2019 in Milan and Turin and 29 January 2020 in Bologna. Virus concentration in the samples ranged from below the limit of detection (LOD) to 5.6 × 104 genome copies (g.c.)/L, and most of the samples (23 out of 26) were below the limit of quantification of PCR. Our results demonstrate that SARS-CoV-2 was already circulating in northern Italy at the end of 2019. Moreover, it was circulating in different geographic regions simultaneously, which changes our previous understanding of the geographical circulation of the virus in Italy. Our study highlights the importance of environmental surveillance as an early warning system, to monitor the levels of virus circulating in the population and identify outbreaks even before cases are notified to the healthcare system

A Reliable and Cost-Efficient PCR-RFLP Tool for the Rapid Identification of Cetaceans in the Mediterranean Sea

Twenty-five species of cetaceans have been reported throughout the Mediterranean Sea, eight of them are commonly distributed in the whole basin and are regularly found beached or adrift in the sea. Stranded animals are frequently found in poor conservation status, preventing reliable identification; identification is thus often based solely on morphological features. Therewith, molecular tools are especially useful to provide taxonomic identification. In this work, a four-enzymes PCR-RFLP in silico protocol, based on a fragment of the mitochondrial gene cytb, has been designed for cetacean species occurring in the Mediterranean Sea. Moreover, beached or floating specimen samples belonging to the eight common species have been tested in the laboratory, providing evidence that this approach represents a reliable, cost- and time-effective tool for their specific identification

First detection of SARS-CoV-2 in untreated wastewaters in Italy

Several studies have demonstrated the advantages of environmental surveillance through the monitoring of sewage for the assessment of viruses circulating in a given community (wastewater-based epidemiology, WBE). During the COVID-19 public health emergency, many reports have described the presence of SARS-CoV-2 RNA in stools from COVID-19 patients, and a few studies reported the occurrence of SARS-CoV-2 in wastewaters worldwide. Italy is among the world's worst-affected countries in the COVID-19 pandemic, but so far there are no studies assessing the presence of SARS-CoV-2 in Italian wastewaters. To this aim, twelve influent sewage samples, collected between February and April 2020 from Wastewater Treatment Plants in Milan and Rome, were tested adapting, for concentration, the standard WHO procedure for Poliovirus surveillance. Molecular analysis was undertaken with three nested protocols, including a newly designed SARS-CoV-2 specific primer set. SARS-CoV-2 RNA detection was accomplished in volumes of 250 ml of wastewaters collected in areas of high (Milan) and low (Rome) epidemic circulation, according to clinical data. Overall, 6 out of 12 samples were positive. One of the positive results was obtained in a Milan wastewater sample collected a few days after the first notified Italian case of autochthonous SARS-CoV-2. The study confirms that WBE has the potential to be applied to SARS-CoV-2 as a sensitive tool to study spatial and temporal trends of virus circulation in the population

Assortative human pair-bonding for partner ancestry and allelic variation of the dopamine receptor D4 (DRD4) gene

The 7R allele of the dopamine receptor D4 gene has been associated with attention-deficit hyperactivity disorder and risk taking. On the cross-population scale, 7R allele frequencies have been shown to be higher in populations with more of a history of long-term migrations. It has also been shown that the 7R allele is associated with individuals having multiple-ancestries. Here we conduct a replication of this latter finding with two independent samples. Measures of subjects’ ancestry are used to examine past reproductive bonds. The individuals’ history of inter-racial/ancestral dating and their feelings about this are also assessed. Tentative support for an association between multiple ancestries and the 7R allele were found. These results are dependent upon the method of questioning subjects about their ancestries. Inter-racial dating and feelings about inter-racial pairing were not related to the presence of the 7R allele. This might be accounted for by secular trends that might have substantively altered the decision-making process employed when considering relationships with individuals from different groups. This study provides continued support for the 7R allele playing a role in migration and/or mate choice patterns. However, replications and extensions of this study are needed and must carefully consider how ancestry/race is assessed

Wastewater surveillance of SARS-CoV-2 variants in October–November 2022 in Italy: detection of XBB.1, BA.2.75 and rapid spread of the BQ.1 lineage

: This study adds insight regarding the occurrence and spread of SARS-CoV-2 Variants of Concern (VOCs) and Variants of Interest (VOIs) in Italy in October and November 2022, by testing urban wastewater collected throughout the country. A total of 332 wastewater samples were collected from 20 Italian Regions/Autonomous Provinces (APs) within the framework of national SARS-CoV-2 environmental surveillance. Of these, 164 were collected in the first week of October and 168 in the first week of November. A ∼1600 bp fragment of the spike protein was sequenced by Sanger (for individual samples) and long-read nanopore sequencing (for pooled Region/AP samples). In October, mutations characteristic of Omicron BA.4/BA.5 were detected in the vast majority (91 %) of the samples amplified by Sanger sequencing. A fraction of these sequences (9 %) also displayed the R346T mutation. Despite the low prevalence documented in clinical cases at the time of sampling, amino acid substitutions characteristic of sublineages BQ.1 or BQ.1.1 were detected in 5 % of sequenced samples from four Regions/APs. A significantly higher variability of sequences and variants was documented in November 2022, when the rate of sequences harbouring mutations of lineages BQ.1 and BQ1.1 increased to 43 %, and the number of Regions/APs positive for the new Omicron subvariant more than tripled (n = 13) compared to October. Moreover, an increase in the number of sequences with the mutation package BA.4/BA.5 + R346T (18 %), as well as the detection of variants never observed before in wastewater in Italy, such as BA.2.75 and XBB.1 (the latter in a Region where no clinical cases associated with this variant had ever been documented) was recorded. The results suggest that, as predicted by the ECDC, BQ.1/BQ.1.1 is rapidly becoming dominant in late 2022. Environmental surveillance proves to be a powerful tool for tracking the spread of SARS-CoV-2 variants/subvariants in the population

Mapping the stranger: genetic diversity of Manila clam in European coastal lagoons.

Manila clam Ruditapes philippinarum - synonym Venerupis philippinarum (Adams and Reeve, 1850) is one of the most successful marine invaders worldwide and represents almost 20% of worldwide mollusc total production. Originally distributed in the Indo-Pacific region, the species was introduced for aquaculture and fisheries in North America, Polynesia, the US Virgin Islands, as well as in Atlantic and Mediterranean coasts of Europe. Due to the commercial value of the species, population genetics and stock composition are of primary importance both in natural and invaded environments. Moreover, molecular genetics can provide valuable data in invasion biology, as, for example, the structure of the exploited populations, the origin of source populations and the routes of invasions. Furthermore, molecular data might be useful for the tracking of seafood products, as requested by the European Commission for European seafood products (see EC laws n° 178/2002, 509/2006, 510/2006 and their improvements). Despite all these considerations genetic data are lacking, especially for introduced populations, and should be urgently provided. In a framework of an European network of researchers involved in Manila clam studies, clams were collected in coastal lagoons and estuaries in both Mediterranean (Northern Adriatic Sea, Italy) and Atlantic (Spain and Portugal) coasts of Europe. Both mitochondrial and nuclear molecular markers were applied to investigate the genetic structure of populations, specifically by the direct sequencing of 16S rDNA gene fragments and by the genotyping of 7 microsatellite markers previously used in native populations. Results demonstrated the occurrence of multiple haplotypes for 16S rDNA, even if one main haplotype was identified in the European analyzed populations. However, few sequences were available in Genbank to make comparisons with native populations. Concerning the microsatellite data, the genotyping of Manila clam populations showed low genetic diversity among them, considering the Fst values, and a lack of genetic structuring when analysed by Structure software. Both these data suggest the absence of geographic differentiation among the introduced populations of Manila clam, as a result of introgression due to translocations and other human activities. The levels of genetic diversity within introduced populations were comparable to those described from native regions, suggesting that multiple introduction events and mixed source populations can counterbalance the loss of genetic diversity caused by the founder effect. However, it must be remarked that microsatellites showed a strong occurrence of null alleles when tested by Micro-checker, as previously observed in other bivalves, suggesting that the problem is differently distributed among Manila clam populations or specific loci, thus microsatellites should be carefully checked before used in extensive population studies. In the near future, population genetics and stock identification of this species on a large scale should be assessed using new powerful markers such as those obtained by NGS (Next Generation Sequencing) methods. In conclusion, this research represents the very first extensive genetic investigation on Manila clam’s European populations, defining the levels of genetic diversity that will help to inform management practices