1,721,027 research outputs found
Two alternative multiplex PCRs for identification of the seven species of anglerfish (Lophius spp) using an end point or a melting curve analysis real-time protocol
No full textAnglerfish (Lophius spp.) is consumed worldwide and is an important economic resource though its seven species are often fraudulently interchanged due to their different commercial value, especially when sold in the form of fillets or pieces. Molecular analysis is the only possible mean to verify traceability and counteract fraud. We developed two multiplex PCRs, one end-point and one real-time with melting curve post-amplification analysis, which can even be run with the simplest two-channel thermocyclers. The two methods were tested on seventy-five reference samples. Their specificity was checked in twenty more species of those most commonly available on the market and in other species of the Lophiidae family. Both methods, the choice of which depends on the equipment and budget of the lab, provide a rapid and easy-to-read response, improving both the simplicity and cost-effectiveness of existing methods for identifying Lophius species
CARATTERIZZAZIONE MOLECOLARE DI PRODOTTI DELLA PESCA D’IMPORTAZIONE CINESE (PESCE GHIACCIO),
No full textTinacci, L., Santoni S., Magni A., Verde F., Armani A. Standardizzazione dei controlli ufficiali sui prodotti della pesca e importati presso il PCF Livorno: implementazione di una checklist operativa. XXXII Convegno Nazionale dell’Associazione Italiana Veterinari Igienisti (AIVI) – “Nuovi approcci della Medicina Veterinaria per uno sviluppo sostenibile di produzioni e consumi alimentari” (Vibo Valentia, 13-15 Settembre 2023).
No full textLe attività di controllo ufficiale presso i posti di Controllo Frontaliero (PCF) sono state recentemente aggiornate ai sensi del Reg. (UE) 2017/625 e del Dlgs 2021/24 con la ridefinizione e l’ampliamento degli ambiti del controllo e delle categorie di prodotto. A tal proposito, si rende necessaria l’implementazione di procedure standardizzate che includano aspetti specifici di controllo per le singole categorie di prodotto. Pertanto, il presente studio è stato finalizzato alla predisposizione di una checklist operativa per il controllo dei prodotti in ingresso presso il PCF Livorno. La predisposizione della checklist è stata preceduta da un’analisi preliminare volta a selezionare: a) la categoria
di prodotti di origine animale oggetto della checklist attraverso la consultazione dei report annuali (2015-2021) relativi ai volumi delle diverse categorie di prodotto per le quali il PCF Livorno è designato al controllo ufficiale; b) le fonti Europee e nazionali inerenti agli aspetti generali delle attività di controllo ufficiale e dei controlli specifici richiesti per la categoria individuata; c) le eventuali checklist preesistenti sviluppate
per il controllo di alimenti di origine animale presso i PCF Italiani. I prodotti ittici sono stati selezionati come categoria target in quanto rappresentano la categoria merceologica maggiormente rappresentata in termini di volume di importazione, sia presso il PCF Livorno che a livello nazionale. Per l’elaborazione degli obiettivi di controllo della nuova checklist sono state incluse 21 fonti normative (17 europee e 4 nazionali) inerenti a requisiti generali e specifici applicati per l’espletazione delle attività di controllo sui prodotti ittici. L’elaborazione della checklist è stata condotta a seguito di un’analisi descrittivo-comparativa di due checklist preesistenti, redatte rispettivamente dal Ministero della Salute e dall’ ex PIF Livorno, rispettivamente in forma di linea guida e documento interno. La comparazione è stata effettuata per procedere alla definizione della struttura della nuova checklist, degli obiettivi dell’attività di controllo sui prodotti ittici ai sensi della normativa vigente e alla selezione di giudizi di valutazione associati per la definizione della ammissibilità delle partite. La completezza, semplicità di utilizzo e agevolazione delle attività di controllo della nuova checklist sono state verificate attraverso la sua applicazione, a campione, per l’esecuzione di controlli su 64 partite ittiche da parte di un operatore in formazione (studente) sotto la supervisione del veterinario ufficiale che aveva partecipato all’elaborazione del documento. Criticità emerse in termini di completezza degli obiettivi sono state risolte prima della definitiva redazione della checklist. In assenza di indicazioni operative europee e nazionali, il presente studio, sviluppato sui prodotti ittici, descrive un approccio potenzialmente utilizzabile per la redazione di checklist relative a tutte le categorie di prodotto per
le quali i PCF sono chiamati ed effettuare i controlli
Residues of veterinary drugs in fish and fish products: An analysis of RASFF data over the last 20 years
Get PDFAn analysis of the notifications in the European Union (EU) Rapid Alert System for Food and Feed (RASFF) portal due to residues of veterinary drugs in fish and fish products over the period 2001–2021 was conducted examining the following data: number and type of notifications; year of notification; notification basis; notifying country; country of origin; action taken; distribution status; risk decision; fish product type; residue found. A total of 292 notifications were found (mean number/year 9.8 ± 13.8 SD), mostly information notifications (60.9%). The most common notification basis was “border control - consignment detained” (38.4%), followed by “official control on the market” (37.9%) and “border control - consignment released” (13.7%). Over half (54.1%) of the notifications were issued by the United Kingdom, Germany and Spain, dominant countries in the fish processing market. Thirty-one countries of origin were recorded, but 48.6% of the notifications were referred to products from Vietnam, followed by China (15.7%), that are among the leading fish producing countries. The most common actions taken were re-dispatch (23.3%), followed by recall from consumer (10.3%), withdrawal from recipients (9.9%), destruction (9.6%), and import not authorised (9.2%). Overall, 28.8% of the notifications involved fish products belonging to the Pangasiidae family (Pangasius spp. and Pangasianodon spp.), followed by tilapia (12.7%), trout (11.0%), eel (8.6%), catfish (7.9%) and salmon (3.4%). Triphenylmethane dyes and their metabolites were the most frequent category of residues found, accounting for 51.4% of the total notifications, followed by a wide range of antibiotics classes, of which nitrofurans and metabolites (19.5%) were the most common, while amphenicols (6.8%), quinolones and fluoroquinolones (6.5%), sulphonamides and potentiators (5.1%), tetracyclines and metabolites (3.4%) and other classes were less represented. Avermectins and unspecified residues were also found in a few cases (2.7%). The annual frequency of these issues varied and was probably influenced by dedicated monitoring plans, as well as by specific sanitary problems occurring in farms. Nevertheless, by providing historical trends and current issues, our analysis identified hazards to be closely monitored, through coordinated official controls especially in the most involved countries
Fish species identification in canned pet food by BLAST and Forensically Informative Nucleotide Sequencing (FINS) analysis of short fragments of the mitochondrial 16s ribosomal RNA gene (16S rRNA)
Get PDFNowadays, pet food claiming high-valued fish among ingredients is largely available on the market. Unfortunately, the modifications induced by processing make species identification by visual inspection difficult and hinder the enforcement of the legislation on traceability. In this work, after aligning 819 sequences of Clupeidae, Engraulidae, Salangidae and Scombridae families, we developed new universal primers for the amplification and sequencing of 2 short fragments (±118 and ±213) of the mitochondrial 16s ribosomal RNA (16S rRNA) gene. Once tested on 130 DNA reference samples, these primers were used in the analysis of highly degraded DNA extracted from 43 canned cat food containing whole minnows (whitebait) (M) and tuna, or bonito or mackerel fillets (F). Three M and 2 F samples were analyzed for each can. A BLAST and a FINS analysis, the latter performed only on the 118 bp fragment, were performed separately on the sequences obtained from M and F samples. All the M samples were identified at the species or genus level by both BLAST and FINS analysis. This allowed to highlight an impressive rate of mislabeling (100%). F samples, for which FINS was less performing in species identification, resulted mislabeled in 40% of the products
Assessment of a Sampling Plan Based on Visual Inspection for the Detection of Anisakid Larvae in Fresh Anchovies (Engraulis encrasicolus). A First Step Towards Official Validation?
Get PDFThe presence of anisakid larvae in fish is a public
health issue, and effective risk management procedures are
needed to avoid that heavily infected products reach the market.
Currently, an official sampling plan for fresh fish defining
sample size, inspection methods, and criteria to accept or reject
the merchandise is lacking at the European and Italian
level. In this study, we compared the visual inspection proposed
by the sampling plan of the Lombardy Region (Italy) to
the UV press method and to an optimized digestion procedure
with the aim to assess its ability in detecting visible parasites.
Thirty-one batches of Engraulis encrasicolus, each composed
of ∼30 specimens, were collected and subsequently analyzed
with the three techniques. The mean abundance (MA) was
calculated after each procedure and compared on the basis of
a threshold value. The results showed that the visual inspection
performed similarly to the digestion method, with a sensitivity
of 93 %, a specificity of 100 %, and an accuracy of
97 %. Overall, the comparison showed that, in the proposed
sampling plan, the visual inspection is effective in rejecting
unmarketable anchovies and in preventing the commercialization
of unsafe products. This method is simple, less demanding
than digestion in terms of time and equipment, and thus suitable as a standardized procedure to be routinely applied by
food business operators. The hazard characterization, performed
by sequencing the mtDNA cox2 gene, has identified
the visible larvae as Anisakis pegreffii in 98 % of the cases,
highlighting the zoonotic potential of the parasites found and
the need for preventive measure
DNA barcoding reveals substitution of Sablefish (Anoplopoma fimbria) with Patagonian and Antarctic Toothfish (Dissostichus eleginoides and D. mawsoni) in online market in China: How mislabeling opens door to IUU fishing
Get PDFChina's rapid economic development has determined profound changes in seafood consumption patterns, and nowadays besides the traditional luxury seafood, high-quality marine fish are consumed. Among these is Anoplopoma fimbria (Sablefish), a highly priced species on the Chinese market. A recent molecular survey on products sold online in China found that all the analyzed products sold as Yin Xue, used to indicate A. fimbria, were instead Dissostichus spp., a genus of fish extremely vulnerable to overfishing (Xiong et al., 2016). Considering this and the lack of a standardized naming system for seafood species in China, an initial search was conducted to identify all the possible Chinese names indicating A. fimbria. The aim of the present study was to assess the challenges of the online market with regards to frauds for fish species substitution. DNA barcoding was employed to verify the identity of 42 products sold on e-commerce platforms as Sablefish. Moreover, the information reported on the webpage and on the label was analyzed according to the Chinese regulation in force. All the PCR products gave readable sequences. By using the IDs analysis on BOLD and the BLAST analysis on GenBank all the samples were unambiguously identified at the species level. Of the 42 products sold as Sablefish, only 6 (14.3%) were molecularly identified as this species, while 32 (76.2%) were identified as Dissostichus eleginoides (Patagonian Toothfish) and 4 (9.5%) as D. mawsoni (Antarctic Toothfish), highlighting an alarming overall misrepresentation rate of 85.7% and implications for the management of these species' fisheries. The combined analysis of all the information of the webpages and the labels allowed us to hypothesize unintentional and intentional mislabeling. Our findings suggest the possible existence of a trade pattern enabling IUU fishing operators to launder illegal catches of Toothfish through mislabeling
A rapid PCR-RFLP method for the identification of Lophius species
No full textThe seven Anglerfish species, which belong to the genus Lophius, have a different value on the market, worldwide. If whole fishes can be identified by their morphological characteristics, they become indistinguishable when prepared or processed. In this study, a rapid method based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was developed for the authentication of the seven Lophius species, using a cytochrome b gene fragment of 566 bp. After a genus-specific PCR, a fast digestion with the restriction enzyme BfaI, followed by agarose gel electrophoresis, allowed a clear species identification by producing specific restriction patterns. The total time required was as low as 6 h, DNA extraction included. The method was then used to analyse 48 commercial samples, whose phylogenetic analysis confirmed the PCR-RFLP response at 100 %. Results showed that mislabelling occurs on the market regardless the kind of processing
Pentaplex PCR as screening assay for jellyfish species identification in food product
Get PDFSalted jellyfish, a traditional food in Asian Countries, is nowadays spreading on the Western markets. In this work, we developed a Pentaplex PCR for the identification of five edible species (Nemopilema nomurai, Rhopilema esculentum, Rhizostoma pulmo, Pelagia noctiluca, and Cotylorhiza tuberculata), which cannot be identified by a mere visual inspection in jellyfish products sold as food. A common degenerated forward primer and five specie-specific reverse primers were designed to amplify COI gene regions of different lengths. Another primer pair targeted the 28SrRNA gene and was intended as common positive reaction control. Considering the high level of degradation in the DNA extracted from acidified and salted products, the maximum length of the amplicons was set at 200 bp. The PCR was developed using 66 reference DNA samples. It gave successful amplifications in 85.4% of 48 ready to eat products (REs) and in 60% of 30 classical salted products (CPs) collected on the market
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