Species identification in petfood by using BLAST analysis of a fragment of the mitochondrial 16s ribosomal RNA gene (16srRNA).

The aim of this work was to verify the label information of pet food for cats, reporting the commercial denomination of Bianchetto among the ingredients, by sequencing and BLAST analysis of a short fragment of the mitochondrial 16SrRNA gene. Fifteen samples of petfood for cats were collected from the retail market. Three fish per samples were analyzed. After DNA extraction, performed according to Armani et al. (2011) (1), the DNA degradation pattern was assessed by electrophoretic analysis. On the basis of the 108 sequences available in GenBank belonging to the order Clupeiformes and Osmeriformes, four different primers were designed and used in combination with those reported in Armani et al. (2012) (2), for the amplification of short fragments of the mitochondrial 16SrRNA gene with a length ranging from 77 to 246 pb. Such fragments were preventively tested by BLAST analysis (http://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastn&BLAST_PROGRAMS=megaBlast&PAGE_TYPE=Bl astSearch&SHOW_DEFAULTS=on&LINK_LOC=blasthome) to assess their discriminatory power at inter and intra specific level. After PCR amplification, the samples associated to the expected amplicon were sequenced and the 45 sequences obtained were analyzed using the program Clustal W in Bioedit version 7.0.9.0 (3) and identified by BLAST analysis. The BLAST analysis returned an identity values of 100% with different species of the genus Encrasicholina. In particular, the most part of the samples were identified at the species level as E. heteroloba, and E. punctifer. The obtained results confirms that the molecular marker selected in this study can be used for the identification of species belonging to the Clupeiformes order, allowing a discrimination even among close species. All the 15 market pet food samples were mislabeled. In fact, while the species identification performed by molecular analysis clearly showed the presence of the juvenile form of Encrasicholina sp, also known as tropical anchovies, all the labels reported the commercial denomination of Bianchetto, which, in Italy, is allowed only for the juvenile form of Sardina pilchardus (4)

Risks and critical issues related to the discovery on the market of unauthorized live alien species on the Italian territory: Chinese crab (Eriocheir sinensis)

Eriocheir sinensis, Chinese Crab or Chinese Mitten Crab is a catadromous species belonging to the Varunidae family, native to river and estuarine areas of North and South East China and Korea. At European level, E. sinensis is widespread in the main water basins of Central and Northern Europe and, since 2016, it has been included in the list of invasive species important for the European Union and subjected to confinement and eradication measures which include the prohibition of collection, transit and placing on the market of live specimens (Regulation (EC) N° 1143/2014). The Chinese Crab can represent a significant danger for the local ecosystem and for the native biota as well as contributing to the appearance of hydrogeological instability phenomena resulting from the intense excavation and erosion of the riverbanks. The first finding of 5 kg of live specimens of Eriocheir sinensis was recorded in the official control by the UFS (Functional Simple Unit) veterinary public health and food safety of the ASL Toscana Centro at an ethnic catering establishment. The specimens were subjected to seizure, photographed, identified morphologically, and subjected to euthanasia and destruction in accordance with the European requirements for welfare and management of animal by-products. From the sanitary point of view, the dangers associated with the consumption of this crab are mainly biological and chemical therefore, risk communication is fundamental, not only at the level of the competent authorities in the sector, but also for the food business operators

Seafood fraud incidents control: Histological based methods as reliable tools for fresh and frozen/thawed products discrimination

Freezing is one of the commonest methods applied for seafood shelf-life extension and long-term preservation. Nevertheless, the quality decay caused by water crystallization and cellular dehydration is responsible of significant reduction of the frozen-thawed products commercial value. Therefore, according to the European Legislation on food labelling, the declaration of freezing process represents a mandatory information for the protection of consumers’ rights. In fact, the lack of such information, besides entailing a non-conformity, can favor commercial frauds. The present work aims to present the development and validation of a histological approach, based on quali-quantitative histological markers, for the discrimination between fresh and thawed-frozen fishery products. In particular, the results of the application of the histological method on Merluccius merluccius and Octopus vulgaris, respectively selected as study models for white meat fish and cephalopods species, will be presented

Safety and commercial issues in fresh mushrooms and mushroom-based products sold at retail in Tuscany region

The compliance to European and National safety and labelling requirements relating to the sale of spontaneous and cultivated mushrooms and mushroom-based products in Tuscany was assessed. The evidence was collected by the Mycological Inspectorate of North-West Tuscany Local Health Authority during 90 inspections (from 2016 to 2020) at large-scale distribution stores, wholesalers, and restaurants in 10 cities belonging to 3 provinces, and on the labelling analysis of 98 commercial products collected at retail in 2021. Despite a substantial compliance of the inspected activities and products with the regulatory requirements, critical issues were highlighted: 1) EU legislative gap in the definition of specific measures for the safe sale of spontaneous mushrooms; 2) improper shelf storage temperatures of fresh-cut products; 3) incorrect condition of use on the labels of pre-packaged products; 4) lack of countryof-origin declaration in pre-packaged products. Furthermore, the labelling analysis highlighted that 18.4% and 15.3% of the products presented issues in the validity and correctness of the scientific names respect to national requirements in. A revision of the current EU legislation is needed to guarantee consumers safety, also considering the relevant number of poisoning cases related to false mycetisms (ingestion of edible mushrooms unproperly stored or used). Also, a specific revision and harmonization of the EU labelling of mushrooms would be desirable to protect consumers

Anisakis spp. larvae in different kinds of ready to eat products made of anchovies (Engraulis encrasicolus) sold in Italian supermarkets

In this study the occurrence of visible anisakid larvae in semi-preserved anchovy products sold on the Italian market was investigated. Totally, 107 ready to eat products (33 salted-ripened, 49 in oil and 25 marinated) were sampled. Each sample was digested, then the digested material was observed under natural and UV light. Parasites were counted, collected and microscopically identified to genus level. A representative subset was molecularly identified using the cox2 gene. At least one visible Anisakis sp. larva was found in 54.2% of the total 107 products analysed and totally 1283 dead larvae were collected. Anisakis sp. larvae were found in all the 33 salted products and 1139 (88.8%) larvae were collected, with a range of 1â105 parasites per product. Larval density per gram was 0.13. Anisakis sp. larvae were found in 49.0% of the products in oil and 143 (11.1%) larvae were isolated, with a range of 0â28 and a density of 0.03. Only 1 larva was found in the 25 marinated products (4.0%, density 0.00). A highly significant difference between all the product categories in respect of number of larvae per product, frequency of products contaminated by at least one larva and larval density per gram was found. Within the subset of larvae molecularly analysed (n = 122), 92 (75.4%) were identified as A. pegreffii and 30 (24.6%) as A. simplex. This study showed that semi-preserved anchovy products heavily contaminated with Anisakis spp. larvae reach the market. Beyond the negligible risk for anisakidosis, the presence of dead visible parasites may cause immediate rejection in consumers. In addition, the potential risk related to allergic reactions in sensitized individuals needs to be further assessed. In order to avoid commercialization of obviously contaminated products, fresh anchoviesâ batches intended for the production of such products should be accurately selected by the processing industry applying inspection methods

An Authentication Survey on Retail Seafood Products Sold on the Bulgarian Market Underlines the Need for Upgrading the Traceability System

Economically motivated or accidental species substitutions lead to economic and potential health damage to consumers with a loss of confidence in the fishery supply chain. In the present study, a three-year survey on 199 retail seafood products sold on the Bulgarian market was addressed to assess: (1) product authenticity by molecular identification; (2) trade name compliance to the list of official trade names accepted in the territory; (3) adherence of the list in force to the market supply. DNA barcoding on mitochondrial and nuclear genes was applied for the identification of whitefish (WF), crustaceans (C) and mollusks (cephalopods-MC; gastropods-MG; bivalves-MB) except for Mytilus sp. products for which the analysis was conducted with a previously validated RFLP PCR protocol. Identification at the species level was obtained for 94.5% of the products. Failures in species allocation were reconducted due to low resolution and reliability or the absence of reference sequences. The study highlighted an overall mislabeling rate of 11%. WF showed the highest mislabeling rate (14%), followed by MB (12.5%), MC (10%) and C (7.9%). This evidence emphasized the use of DNA-based methods as tools for seafood authentication. The presence of non-compliant trade names and the ineffectiveness of the list to describe the market species varieties attested to the need to improve seafood labeling and traceability at the national level

Selection of Histological Parameters for the Development of an Analytical Method for Discriminating Fresh and Frozen/Thawed Common Octopus (Octopus vulgaris) and Preventing Frauds along the Seafood Chain

This study aimed at selecting effective histological indicators of the freezing process, for the discrimination of fresh and frozen common octopus Octopus vulgaris. Histological indices of freezing process were selected in mantle and arm muscle and axial nerve tissues. Seven histological parameters were chosen: overall muscle tissue structural organization (a); gaping among muscle bundles (b); presence of optically empty spaces between and within muscle bundles (c); white spaces percentage between and within muscle bundles (d); overall nerve structural organization (e); presence of linear fissures and/or empty spaces within neuropil and axonal tract (f); presence of empty spaces within connective tissue matrix surrounding the nerve (g); empty space percentage within arm axial nerve region (h). The parameters were assessed on 150 mantle muscle sections (a, b, c, d), 150 arm muscle (b, c) and nervous (e, f, g, h) tissue sections belonging to 20 fresh exemplars further subdued to conventional freezing procedure at − 20 °C, 25 fresh curled exemplars, 25 exemplars industrially frozen at − 80 °C, and 20 thawed, curled, and individually quick frozen exemplars. Overall structural organization (a, e), gaping (b), and presence of optically empty spaces (c, f) confirmed significant morphological freezing indices in mantle and arm sections. In mantle, two d values (d < 25% and 25% < d < 33%) were proposed as complementary thresholds to be applied in association with b and c indices for freezing discrimination. These parameters appear eligible to set a method for discriminating fresh/thawed octopus products to be applied both in official control and self-check activities

what happened in china after the melamin crisis

Recently, many changes have been introduced in the food safety control organization of the People's Republic of China, pushed by important international events to be held within the Country and some big scandals, such as the "melamine milk crisis". The old "law on food hygiene" of 1995 has been replaced by the "law for food safety" in 2009, to reorganize in particular, the arrangement and responsibilities of the Institutions that operate in this field. In particular, the National Council for Food Safety has been created. It represents the highest authority in the matter, for legislation and investigation on food concerns and coordination of the aforesaid Institutions. However, notwithstanding the many steps ahead, many efforts remain to be performed especially in terms of formation of the food business operators and to solve complex problems, such as the big fragmentation of the primary production, which, at present, would need a huge extent of human and financial resources for the control system