Integrated DNA barcoding methods to identify species in the processed fish products from Chinese market.

DNA-based methods are reliable for a precise identification of species in processed products. In this study, we assessed five typical DNA extraction methods from multiple aspects. Full-length and mini-length DNA barcoding were performed to detect the species substitution and mislabeling of 305 processed fish products from the Chinese market covering six processed fish products. The salt extraction method that exhibited the best overall performance was applied. All samples were successfully extracted; however, only 19.3 % of samples could be amplified using the full-DNA barcode primer set, and 90.2 % of samples could be amplified using the newly designed mini-DNA barcode primer sets (401 and 320 bp). Overall, the molecular identification results revealed that 36.4 % (111/305) of the samples were inconsistent with the labels, with commercial fraud observed in all six types of processed fish products. The survey findings provide technical references for effective fish authentication monitoring, offering insights into the seafood safety in markets.

Re-visiting the occurrence of mislabeling in frozen "pescada-branca" (Cynoscion leiarchus and Plagioscion squamosissimus - Sciaenidae) sold in Brazil using DNA barcoding and octaplex PCR assay.

In Brazil, Cynoscion leiarchus and Plagioscion squamosissimus are the species allowed to be labeled as "pescada-branca". These species have high economic value, especially when sold in the form of fillets. Therefore, when morphological traits are removed, fish are highly prone to be substituted, which has been reported for species of the family Sciaenidae sold in Brazil, including "pescada-branca". We have sequenced 618 bp of the COI of 143 samples to re-evaluate the occurrence of substitutions in frozen "pescada-branca" marketed in Brazil. We observed more than 73% of mislabeling, with only 26.57% being P. squamosissimus, and none, C. leiarchus. In general, the substitutes were closely related Sciaenidae, but cheaper species, which indicates commercial fraud. Based on these results we used 1.2 kb of COI to develop an octaplex PCR assay that unequivocally identified the target species and six substitute species through the banding pattern. Specific reverse primers combined with a universal forward primer were used in the protocol and identified the species C. leiarchus (~290 bp), N. microps (~340 bp), M. ancylodon (~470 bp), C. acoupa (~540 bp), C. microlepidotus (~850 bp), P. auratus (~950 bp), C. virescens (~1050 bp), and P. squamosissimus (~1140 bp). The DNA barcoding and the multiplex PCR were accurate and specific to authenticate processed products labeled as "pescada-branca". The multiplex assay constitutes a cost-effective alternative for the authentication of these products and other sciaenids. Additionally, we suggest that the multiplex assay can be adopted by both companies and regulatory agencies to prevent commercial fraud in the marketing of processed fishery products in Brazil and other countries where these products are commercialized.

Governmental actions to address COVID-19 misinformation.

Since COVID-19 emerged, a plethora of misinformation has undermined the public's ability to identify reliable sources of accurate information. To identify the range of methods governments used to address COVID-19 misinformation, we conducted a content analysis of international media and evaluated government actions in light of international law, which protects freedom of expression and calls on governments to guarantee this fundamental right even during a pandemic or other emergency. We identified five categories of government activities: (1) disseminating and increasing access to accurate information; (2) restricting access to accurate information; (3) disseminating disinformation, false information, and misinformation; (4) addressing commercial fraud; and (5) criminalizing expression. The goal of addressing COVID-19 misinformation is best served by protecting expression, disseminating factual information, ensuring strong protections for whistleblowers, and supporting an independent media environment. Conversely, governments undermine public health when they create a state of uncertainty and violate human rights.

DNA barcoding reveals mislabeling and commercial fraud in the marketing of fillets of the genus Brachyplatystoma Bleeker, 1862, the Amazonian freshwater catfishes economically important in Brazil.

The substitution and mislabeling is facilitated by the processing of fish products. We employed a DNA barcoding to authenticate fillets labeled as "dourada" (Brachyplatystoma rousseauxii), and "piramutaba" (Brachyplatystoma vaillantii) marketed in the Brazil. A 615 bp of the Cytochrome oxidase subunit I (COI) was sequenced from 305 fillets and subsequently identified to species level by querying public databases and sequences of reference species. The results revealed a global mean substitution rate of 17%. The highest substitution rate was detected in "dourada" (26%), the most valuable species, followed by "piramutaba" (9%). The most cases of substitutions were by species of lower commercial value, suggesting fraud aimed at increased profits. Therefore, we suggest the improvement of food-labeling regulation, continued inspection, as well as the adoption of the DNA barcode for the molecular authentication of processed fish to prevent substitution of these products in Brazil.

Target vs spectral fingerprint data analysis of Iberian ham samples for avoiding labelling fraud using headspace - gas chromatography-ion mobility spectrometry.

The data obtained with a polar or non-polar gas chromatography (GC) column coupled to ion mobility spectrometry (IMS) has been explored to classify Iberian ham, to detect possible frauds in their labelling. GC-IMS was used to detect the volatile compound profile of dry-cured Iberian ham from pigs fattened on acorn and pasture or on feed. Due to the two-dimensional nature of GC-IMS measurements, great quantities of data are obtained and an exhaustive chemometric processing is required. A first approach was based on the processing of the complete spectral fingerprint, while the second consisted of the selection of individual markers that appeared throughout the spectra. A classification rate of 90% was obtained with the first strategy, and the second approach correctly classified all Iberian ham samples according to the pigs' diet (classification rate of 100%). No significant differences were found between the GC columns tested in terms of classification rate.

Molecular authentication of Pargo fillets Lutjanus purpureus (Perciformes: Lutjanidae) by DNA barcoding reveals commercial fraud

The Caribbean Red Snapper (Pargo) Lutjanus purpureus is the most economically important snapper in Brazil, which is sold, among other forms, as frozen fillets. During the process of transformation into fillets there is the removal of the distinctive morphological traits, being able to favor the substitution by less valued species. In addition, there is no national legislation requiring the insertion of the specific name on the product label. However, according to a Normative Instruction (IN N ° 29/2015 MAPA) that correlates the common and specific names of the products destined to the national trade, in Brazil only L. purpureus and L. campechanus can be denominated "Pargo". Thus, the DNA barcode tool was used to identify the fillets sold in north of Brazil, labeled "Pargo", with the aid of sequences from the public and control databases. The results showed that among 142 fillets examined, 78% was identified as L. purpureus and 22% as Rhomboplites aurorubens, a snapper with low commercial value in the country, revealing commercial fraud. The molecular identification method successfully used in this study to authenticate fillets snappers may also be used by surveillance authorities in the quality control of processed fish products, towards ensuring consumer rights.(AU)

O Pargo Lutjanus purpureus, lutjanídeo mais importante economicamente no Brasil, é vendido, entre outras formas, como filés congelados. Durante a transformação em filés, há a remoção das características morfológicas distintivas, podendo favorecer a substituição por espécies menos valorizadas. Além disso, não há legislação nacional que exija a inserção do nome específico no rótulo. Porém, de acordo com uma Instrução Normativa (IN N° 29 /2015 MAPA) que correlaciona os nomes comuns e específicos dos produtos destinados ao comércio nacional, no Brasil somente L. purpureus e L. campechanus podem ser denominados "Pargo". Assim, a ferramenta DNA barcode foi usada para identificar os filés vendidos no norte do Brasil, rotulados como "Pargo", com o auxílio de sequências dos bancos de dados públicos e banco controle. Os resultados mostraram que entre os 142 filés examinados, 78% foi identificado como L. purpureus e 22% como Rhomboplites aurorubens, um lutjanídeo com baixo valor comercial no país, revelando fraude comercial. O método de identificação molecular, utilizado com êxito neste estudo para autenticar filés de lutjanídeos, pode também ser utilizado pelas autoridades de vigilância no controle de qualidade de produtos processados derivados de peixes em geral, para garantir os direitos dos consumidores.(AU)

Pyrosequencing as a tool for rapid fish species identification and commercial fraud detection.

The increased consumption of fish products, as well as the occurrence of exotic fish species in the Mediterranean Sea and in the fish market, has increased the risk of commercial fraud. Furthermore, the great amount of processed seafood products has greatly limited the application of classic identification systems. DNA-based identification allows a clear and unambiguous detection of polymorphisms between species, permitting differentiation and identification of both commercial fraud and introduction of species with potential toxic effects on humans. In this study, a novel DNA-based approach for differentiation of fish species based on pyrosequencing technology has been developed. Raw and processed fish products were tested, and up to 25 species of fish belonging to Clupeiformes and Pleuronectiformes groups were uniquely and rapidly identified. The proper identification based on short and unique genetic sequence signatures demonstrates that this approach is promising and cost-effective for large-scale surveys.

Effect of freezing/thawing process in different sizes of blue fish in the Mediterranean through lysosomal enzymatic tests.

The assessment of freshness of different sizes of blue fish (Engraulis encrasicolus 12 cm, Sardina pilchardus 15 cm, Trachurus trachurus 40 cm, Scomber japonicus colias 60 cm) was carried out using non-conventional enzymatic methods. The activities of the three lysosomal enzymes (α-glucosidase (AG), ß-galactosidase (B-GAL) and ß-N-acetylglucosamidase (B-NA)) in extracts of blue fish muscle were measured over a period of 21 days of storage. A significant increase (p<0.05) of AG activity was observed in all species, with a large increase seen after only one day of storage. B-NA activity increased slightly in sardines, horse mackerels and chub mackerel during frozen/thawed storage. Finally, the increase of B-GAL activity was significant (p<0.05) only in the samples of larger blue fish as horse mackerel and chub mackerel. All of these enzyme activities may be helpful predictive markers to limit fraud in these species.

Avaliação das perdas de líquido por degelo de frangos congelados (Drip Test) em abatedouros / Drip losses evaluation in frozen chickens (Drip Test) at abattoirs

Este trabalho objetivou avaliar a quantidade de água resultante do descongelamento de carcaças de aves oriundas de quatro abatedouros de aves no Estado de São Paulo, sob Serviço de Inspeção Estadual, através do método Drip Test, durante os anos de 2011 e 2012. Foram utilizadas 45 amostras, sendo que cada uma dessas era composta por seis carcaças de frango congeladas, determinando um total de 270 carcaças de aves utilizadas. Durante a realização do estudo, apenas 4,44% (2/45) das amostras apresentaram valores acima do limite aceitável determinado pelo Ministério da Agricultura, Pecuária e Abastecimento (MAPA), sendo que essas pertenciam a abatedouros distintos, porém não foi observada diferença significativa entre todos os estabelecimentos avaliados. Pode-se concluir que 4,44% das amostras do monitoramento apresentaram-se fora do padrão legal, oriundas de falhas tecnológicas e que resultam na insatisfação dos consumidores. É necessário um monitoramento de rotina, com colheita de amostras prontas para a comercialização sem prévio aviso, como prática a salvaguardar os interesses dos consumidores e à coibição de fraudes intencionais, principalmente naqueles estabelecimentos que evidenciem valores frequentemente elevados.

This study aimed to evaluate the amount of water resulting from poultry carcasses from four slaughterhouses at the São Paulo State, Brazil, through “Drip Test” method, during the years 2011 and 2012. It was used 45 samples, which were consisted of 6 chickenfrozen carcasses, totalizing 270 chickens. During the study, 4.44% (2/45) of the samples had values above the acceptable limit by Ministry of Agriculture, Livestock and Supply (MAPA), and were from two different establishments, but was not observed significant difference among all the establishments. It was concluded that some samples were not in concordance with legal standards, arising from technological failures that contributes to cause consumer dissatisfaction. Is necessary a routine monitoring system, utilizing samples ready for commercialization without previously notice, to safeguard the consumer’s interests and avoid intentional fraud, especially in those establishments that commonly presents high values.

Myoglobin as marker in meat adulteration: a UPLC method for determining the presence of pork meat in raw beef burger.

The identification of meat animal species used in raw burgers is very important with respect to economic and religious considerations. Therefore, international supervisory bodies have implemented procedures to control the employed meat species. In this paper we propose myoglobin as a powerful molecular marker to evaluate the presence of non-declared meat addition in raw beef burgers by using ultra-performance liquid chromatography (UPLC) for the separation and identification of edible animal species (beef, chicken, horse, ostrich, pig and water buffalo). Meat samples were pre-treated with sodium nitrite to transform oxymyoglobin and deoxymyoglobin to the more stable metmyoglobin. The developed method was validated, preparing mixtures with different percentages of pork and beef minced meat. The obtained results show that using myoglobin as marker, 5% (25 mg/500 mg) of pork or beef meat can be detected in premixed minced meat samples.