1,721,000 research outputs found
Development of suitable tools to produce a DISC vaccine to control African Swine Fever
Get PDFASF is a highly contagious, lethal, and economically devasting infectious disease that affects bush pigs, warthogs, domestic pigs, and wild boars. It is present in Africa, eastern and western Europe, Asia, and Sardinia where it is endemic since 1978. The etiological agent is the ASFV, a large, enveloped icosahedral virus with a dsDNA genome. The lack of a vaccine limits the options to control the disease. For this reason, it is important the development of a vaccine capable of infecting and inducing a long last immunity, avoiding the arise of side effects and the risk of possible recombination with the wild type virus to eradicate the disease. The main goal of the project is the development of a DISC vaccine specific for ASFV Sardinian isolates. Vaccines production requires the development of suitable continuous porcine cell lines genetically stable to support viral replication. The natural target cells for ASFV are cells derived from monocytes, in particular macrophages. At present, there are not monocyte/macrophage continuous cell lines permissive to virus replication. In the natural host, ASFV replicates in endothelial cells as well. This work showed the development of an immortalized Porcine Aortic Endothelial Cell (iPAEC) line using E6-E7 oncogenes derived from OaPV3 ovine papillomavirus. This allowed us to obtain a suitable continuous cell line that may be used to develop a DISC vaccine. Analysis on iPAEC cells showed that they are capable to support viral replication revealing a highly susceptibility to ASFV infection. In addition, the production of a DISC vaccine requires the deletion of selected ASFV genes. This project showed the design of synthetical target late genes and their expression in Cos-1 cells. The presented work allowed us to set up a strategy to develop helper cell lines expressing mutagenized genes and capable to support replication of defective viruses. The methodology here described represents a new suitable tool for vaccines development.African Swine Fever is a highly contagious, lethal, and economically devasting infectious disease that affects bush pigs, warthogs, domestic pigs, and wild boars. ASF is present in Africa, eastern and western Europe, Asia, and Sardinia, where it is endemic since 1978. The etiological agent of ASF is the African Swine Fever Virus (ASFV), a large, enveloped virus assembled by an icosahedral structure containing a double-strand DNA genome. The lack of a vaccine limits the options to control the disease. For this reason, it is important the development of a vaccine capable of infecting and inducing a long last immunity as well as avoiding the arise of side effects and the risk of possible recombination with the wild type virus to eradicate the disease. Therefore, the main goal of the present project is the development of a DISC vaccine specific for ASFV Sardinian isolates. Vaccines’ production requires the development of suitable continuous porcine cell lines genetically stable to support viral replication. The natural target cells for ASFV are cells derived from monocytes, in particular macrophages. At present, there are not monocyte/macrophage continuous cell lines permissive to virus replication. In the natural host, ASFV replicates in endothelial cells as well. This work showed the development of an immortalized Porcine Aortic Endothelial Cell (iPAEC) line using E6-E7 oncogenes derived from OaPV3 ovine papillomavirus. This allowed us to obtain a suitable continuous cell line derived from porcine that may be used to develop a DISC vaccine. Analysis on iPAEC cells showed that they are capable to support viral replication revealing a highly susceptibility to ASFV infection. In addition, the production of a DISC vaccine requires the deletion of selected ASFV genes. This project showed the design of synthetical target late genes and their expression in Cos-1 cell lines. The presented work allowed us to set up a strategy to develop helper cell lines expressing mutagenized genes and capable to support replication of defective viruses. The methodology here described represents a new suitable tool for vaccines development
Diatom diversity through HTS-metabarcoding in coastal European seas
No full textPiredda, Roberta ... et al.-- 12 pages, 5 figures, 1 table, supporting information https://doi.org/10.1038/s41598-018-36345-9Diatoms constitute a diverse lineage of unicellular organisms abundant and ecologically important in aquatic ecosystems. Compared to other protists, their biology and taxonomy are well-studied, offering the opportunity to combine traditional approaches and new technologies. We examined a dataset of diatom 18S rRNA- and rDNA- (V4 region) reads from different plankton size-fractions and sediments from six European coastal marine sites, with the aim of identifying peculiarities and commonalities with respect to the whole protistan community. Almost all metabarcodes (99.6%) were assigned to known genera (121) and species (236), the most abundant of which were those already known from classic studies and coincided with those seen in light microscopy. rDNA and rRNA showed comparable patterns for the dominant taxa, but rRNA revealed a much higher diversity particularly in the sediment communities. Peculiar to diatoms is a tight bentho-pelagic coupling, with many benthic or planktonic species colonizing both water column and sediments and the dominance of planktonic species in both habitats. Overall metabarcoding results reflected the marked specificity of diatoms compared to other protistan groups in terms of morphological and ecological characteristics, at the same time confirming their great potential in the description of protist communitiesFinancial support to the project BioMarKs was provided by the EU (2008-6530, ERA-net BIODIVERSA, EU) and, for the Italian participation, by Stazione Zoologica Anton Dohrn and the projects FIRB Biodiversitalia-Approccio multitaxa allo studio delle risposte della biodiversità italiana al cambiamento climatico (RBAP10A2T4), funded by the Italian Ministry of Education, University and Research (MIUR), which also supported Roberta Piredda’s fellowshipPeer Reviewe
Authentication of seafood species on the ASFIS list (FAO) by in-silico evaluation of primers for metabarcoding
Get PDFGiven the remarkable biodiversity among the seafood species entering the food supply chain, authentication is a crucial component in ensuring transparent trade. DNA metabarcoding stands out as the leading methodology for determining species composition within complex food matrices. Nevertheless, the selection of primer pairs is critical, as it may introduce biases into species identification. In this investigation, we evaluated previously described primers for their in-silico efficacy in identifying both edible marine species on the ASFIS (FAO) list and the main terrestrial animal species frequently incorporated as ingredients in processed seafood products. Selected
primers covered the Cytochrome c Oxidase subunit I (COI), the Cytochrome b (cytb), the 16S rRNA (16S) and the 12S rRNA (12S) mitochondrial regions. Results showed that, out of 53 pairs analysed, one primer pair targeting the COI region obtained the best performance in terms of taxonomic coverage for both ASFIS and terrestrial species. Notably, each primer pair showed a unique taxonomic profile, suggesting their potential for addressing various seafood authentication issues as the detection of novel food ingredients (e.g., jellyfish, red algae, insects) or endangered species. Overall, these outcomes deliver valuable insights for selecting metabarcoding primers to improve the traceability of seafood products and contribute to the development of an innovative Food Safety Management System
Species substitution in goat yoghurt supply chain using melting-curve analysis
No full textMislabelling of dairy products has been widely reported worldwide but there are relatively few data available to help assess species substitution in goat yogurt. The purpose of this study was to investigate the presence of bovine and sheep milk in yogurt labeled as being made exclusively from goat milk, using qualitative real-time PCR. The results confirm that fermented products are targets for food species substitution, highlighting the undeclared presence of cow and/or sheep milk in 39.6% of samples analysed. Moreover, the study shows that the qualitative real-time PCR used here is a rapid, sensitive (LODabs of 0.015 ng/mu L), specific (no primer-dimers or non-specific products in the reactions), straightforward and low-cost molecular tool for discriminating adulterants in the dairy supply chain. Therefore, melt-curve analysis is a promising tool for facilitating routine safety, quality and authenticity checks in the dairy industry, and for implementing mitigation strategies against fraudulent activities. Application of DNA-based approaches is crucial for designing an innovative food safety management system and for helping to implement both a Food Fraud Vulnerability Assessment and a Food Fraud Prevention Strategy as common practices in the dairy sector
Occurrence of hepatitis E virus (HEV) in Calabrian wild boars
Get PDFHepatitis E virus (HEV) is an emerging pathogen in industrialized countries. HEV infections in humans are mainly related to the HEV-3 genotype, predominant in Europe and widespread in wild boars' food products. However, there are little relevant data around HEV prevalence in wild boars, although they are considered the main HEV reservoir and used for typical food products such as liver sausages. Our study aimed to assess HEV occurrence and genetic variability in Calabrian wild boars hunted in the central and ionic area of Catanzaro's province. A total of 86 wild boar liver samples were analyzed showing an overall HEV RNA prevalence of 26.7% (23/86). All positive samples were characterized molecularly as genotype 3 and predicted as HEV-3c subtype despite the shortness of fragment employed for the molecular analysis. This data is in line with previous studies conducted in Europe highlighting the public health concern of these results. Biomolecular methods performed in our study detected only the HEV RNA positivity of analyzed samples without information about the virus viability. Consequently, it is not possible to fully estimate the risk related to the consumption of wild boar's liver sausages or wild boar meat products. Our results highlight the need for further studies in order to investigate the virus viability and to link wild boar's meat consumption with HEV human seroprevalence in Italian regions (Abruzzo, Lazio, Campania and Calabria) where typical wild boar's products are consumed. In this way, the Competent Authorities could perform a complete risk assessment, implement risk management and establish proper measures to ensure the public health and prevent relative human disease.: Hepatitis E virus (HEV) is an emerging pathogen in industrialized countries. HEV infections in humans are mainly related to the HEV-3 genotype, predominant in Europe and widespread in wild boars' food products. However, there are little relevant data around HEV prevalence in wild boars, although they are considered the main HEV reservoir and used for typical food products such as liver sausages. Our study aimed to assess HEV occurrence and genetic variability in Calabrian wild boars hunted in the central and ionic area of Catanzaro's province. A total of 86 wild boar liver samples were analyzed showing an overall HEV RNA prevalence of 26.7% (23/86). All positive samples were characterized molecularly as genotype 3 and predicted as HEV-3c subtype despite the shortness of fragment employed for the molecular analysis. This data is in line with previous studies conducted in Europe highlighting the public health concern of these results. Biomolecular methods performed in our study detected only the HEV RNA positivity of analyzed samples without information about the virus viability. Consequently, it is not possible to fully estimate the risk related to the consumption of wild boar's liver sausages or wild boar meat products. Our results highlight the need for further studies in order to investigate the virus viability and to link wild boar's meat consumption with HEV human seroprevalence in Italian regions (Abruzzo, Lazio, Campania and Calabria) where typical wild boar's products are consumed. In this way, the Competent Authorities could perform a complete risk assessment, implement risk management and establish proper measures to ensure the public health and prevent relative human disease
Authentication of the Botanical Origin of Honey: In Silico Assessment of Primers for DNA Metabarcoding
No full textHoney is a valuable food product that is often mislabeled as to its botanical composition, affecting both trade transparency and consumer safety. DNA metabarcoding is a promising method for authenticity verification, although the results are affected by the choice of molecular markers and primer pairs. This study selected 43 primer pairs from the literature covering several genes (chloroplast rbcL, matK, and trnL and nuclear ITS) and length of <600 bp. Performance of these primers was in silico assessed for all Streptophyta and a subset of them representing the honeybee foraging plants included in the European list. The results showed that rbcL primers provided the highest taxonomic coverage, which increased significantly when combined with ITS2,
especially for key families, such as Asteraceae, Fabaceae, Lamiaceae, and Rosaceae. These results provide valuable insights for the design and chemical setup of molecular steps in metabarcoding experiments applied to honey authenticity also supporting the standardization of official control protocols
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Get PDF“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Get PDFWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
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