Fraud in highly appreciated fish detected from DNA in Europe may undermine the Development Goal of sustainable fishing in Africa.

Despite high effort for food traceability to ensure safe and sustainable consumption, mislabeling persists on seafood markets. Determining what drives deliberate fraud is necessary to improve food authenticity and sustainability. In this study, the relationship between consumer's appreciation and fraudulent mislabeling was assessed through a combination of a survey on consumer's preferences (N = 1608) and molecular tools applied to fish samples commercialized by European companies. We analyzed 401 samples of fish highly consumed in Europe and worldwide (i.e. tuna, hake, anchovy, and blue whiting) through PCR-amplification and sequencing of a suite of DNA markers. Results revealed low mislabeling rate (1.9%), with a higher mislabeling risk in non-recognizable products and significant mediation of fish price between consumer´s appreciation and mislabeling risk of a species. Furthermore, the use of endangered species (e.g. Thunnus thynnus), tuna juveniles for anchovy, and still not regulated Merluccius polli hake as substitutes, points towards illegal, unreported and/or unregulated fishing from African waters. These findings reveal a worrying intentional fraud that hampers the goal of sustainable seafood production and consumption, and suggest to prioritize control efforts on highly appreciated species.

An extremely sensitive nested PCR-RFLP mitochondrial marker for detection and identification of salmonids in eDNA from water samples.

BACKGROUND: Salmonids are native from the North Hemisphere but have been introduced for aquaculture and sport fishing in the South Hemisphere and inhabit most rivers and lakes in temperate and cold regions worldwide. Five species are included in the Global Invasive Species Database: rainbow trout Oncorhynchus mykiss, Atlantic salmon Salmo salar, brown trout Salmo trutta, brook trout Salvelinus fontinalis, and lake trout Salvelinus namaycush. In contrast, other salmonids are endangered in their native settings. METHODS: Here we have developed a method to identify salmonid species directly from water samples, focusing on the Iberian Peninsula as a case study. We have designed nested Salmonidae-specific primers within the 16S rDNA region. From these primers and a PCR-RFLP procedure the target species can be unequivocally identified from DNA extracted from water samples. RESULTS: The method was validated in aquarium experiments and in the field with water from watersheds with known salmonid populations. Finally, the method was applied to obtain a global view of the Salmonidae community in Nalón River (north coast of Spain). DISCUSSION: This new powerful, very sensitive (identifying the species down to 10 pg DNA/ml water) and economical tool can be applied for monitoring the presence of salmonids in a variety of situations, from checking upstream colonization after removal of river barriers to monitoring potential escapes from fish farms.

NGS tools for traceability in candies as high processed food products: Ion Torrent PGM versus conventional PCR-cloning.

The Next Generation Sequencing methodologies are considered the next step within DNA-based methods and their applicability in different fields is being evaluated. Here, we tested the usefulness of the Ion Torrent Personal Genome Machine (PGM) in food traceability analyzing candies as a model of high processed foods, and compared the results with those obtained by PCR-cloning-sequencing (PCR-CS). The majority of samples exhibited consistency between methodologies, yielding more information and species per product from the PGM platform than PCR-CS. Significantly higher AT-content in sequences of the same species was also obtained from PGM. This together with some taxonomical discrepancies between methodologies suggest that the PGM platform is still pre-mature for its use in food traceability of complex highly processed products. It could be a good option for analysis of less complex food, saving time and cost per sample.

Detection of Different DNA Animal Species in Commercial Candy Products.

Candy products are consumed all across the world, but there is not much information about their composition. In this study we have used a DNA-based approach for determining the animal species occurring in 40 commercial candies of different types. We extracted DNA and performed PCR amplification, cloning and sequencing for obtaining species-informative DNA sequences. Eight species were identified including fish (hake and anchovy) in 22% of the products analyzed. Bovine and porcine were the most abundant appearing in 27 samples each one. Most products contained a mixture of species. Marshmallows (7), jelly-types, and gummies (20) contained a significantly higher number of species than hard candies (9). We demonstrated the presence of DNA animal species in candy product which allow consumers to make choices and prevent allergic reaction.

Authentication of commercial candy ingredients using DNA PCR-cloning methodology.

BACKGROUND: Commercial candies are consumed by all population age sectors worldwide. Methods for quality control and composition authentication are therefore needed for best compliance with consumers' preferences. In this study applications of DNA-based methodology for candy quality control have been tested. Eighteen samples of commercial candies (marshmallows, gumdrops, jelly, sherbet, gelatin-based desserts) produced by five countries were analyzed to identify the component species by polymerase chain reaction, cloning and sequencing of 16S rRNA and ribulose -1,5-diphosphate carboxylase oxygenase genes, and the species determined from BLAST comparison with universal databases and phylogenetic analysis. RESULTS: Positive DNA extraction and amplification of the target genes were obtained for 94% of candies assayed, even those containing as little as <0.0005 ng µL(-1) DNA concentration. The results demonstrated that the species detected from DNA were compatible with the information provided on candy labels only in a few products. DNA traces of undeclared species, including fish, were found in most samples, and two products were labeled as vegetarian but contained porcine DNA. CONCLUSION: Based on the inaccuracy found on the labels of sweets we recommend the use of DNA tests for quality control of these popular sweets. DNA tests have been useful in this field but next-generation sequencing methods could be more effective.

Detecting nuisance species using NGST: Methodology shortcomings and possible application in ballast water monitoring.

Detecting the presence of potential invasive species in ballast water is a priority for preventing their spread into new environments. Next generation sequencing technologies are being increasingly used for exploring and assessing biodiversity from environmental samples. Here we apply high throughput sequencing from DNA extracted from ballast water (BW) samples employing two different platforms, Ion Torrent and 454, and compare the putative species catalogues from the resulting Operational Taxonomic Units (OTU). Water samples were taken from the RV Polastern ballast tank in five different days between the second and the twentieth navigation day. Pronounced decrease of oxygen concentration and increase of temperature occurred in the BW during this time, coincident with a progressively higher proportion of unassigned OTU and short reads indicating DNA degradation. Discrepancy between platforms for species catalogues was consistent with previously published bias in AT-rich sequences for Ion Torrent platform. Some putative species detected from the two platforms increased in frequency during the Polarstern travel, which suggests they were alive and therefore tolerant to adverse conditions. OTU assigned to the highly invasive red alga Polysiphonia have been detected at low but increasing frequency from the two platforms. Although in this moment NGST could not replace current methods of sampling, sorting and individual taxonomic identification of BW biota, it has potential as an exploratory methodology especially for detecting scarce species.

Over the sands and far away: interpreting an Iberian mitochondrial lineage with ancient Western African origins.

OBJECTIVES: There is an ongoing effort to characterize the genetic links between Africa and Europe, mostly using lineages and haplotypes that are specific to one continent but had an ancient origin in the other. Mitochondrial DNA has been proven to be a very useful tool for this purpose since a high number of putatively European-specific variants of the African L* lineages have been defined over the years. Due to their geographic locations, Spain and Portugal seem to be ideal places for searching for these lineages. METHODS: Five members of a minor branch of haplogroup L3f were found in recent DNA samplings in the region of Asturias (Northern Spain), which is known for its historical isolation. The frequency of L3f in this population (≈1%) is unexpectedly high in comparison with other related lineages in Europe. Complete mitochondrial DNA sequencing of these L3f lineages, as well phylogenetic and phylogeographic comparative analyses have been performed. RESULTS: The L3f variant found in Asturias seems to constitute an Iberian-specific haplogroup, distantly related to lineages in Northern Africa and with a deep ancestry in Western Africa. Coalescent algorithms estimate the minimum arrival time as 8,000 years ago, and a possible route through the Gibraltar Strait. CONCLUSIONS: Results are concordant with a previously proposed Neolithic connection between Southern Europe and Western Africa, which might be key to the proper understanding of the ancient links between these two continents.

Evaluation of large-scale genetic structure in complex demographic and historical scenarios: the mitochondrial DNA and Y-chromosome pools of the Iberian Atlantic façade.

Genetic structural patterns of human populations are usually a combination of long-term evolutionary forces and short-term social, cultural, and demographic processes. Recently, using mitochondrial DNA and Y-chromosome loci, various studies in northern Spain have found evidence that the geographical distribution of Iron Age tribal peoples might have influenced current patterns of genetic structuring in several autochthonous populations. Using the wealth of data that are currently available from the whole territory of the Iberian Peninsula, we have evaluated its genetic structuring in the spatial scale of the Atlantic façade. Hierarchical tree modeling procedures, combined with a classic analysis of molecular variance (AMOVA), were used to model known sociocultural divisions from the third century BCE to the eighth century CE, contrasting them with uniparental marker data. Our results show that, while mountainous and abrupt areas of the Iberian North bear the signals of long-term isolation in their maternal and paternal gene pools, the makeup of the Atlantic façade as a whole can be related to tribal population groups that predate the Roman conquest of the Peninsula. The maintenance through time of such a structure can be related to the numerous geographic barriers of the Iberian mainland, which have historically conditioned its settlement patterns and the occurrence of genetic drift processes.

Assessing the genetic influence of ancient sociopolitical structure: micro-differentiation patterns in the population of Asturias (Northern Spain).

The human populations of the Iberian Peninsula are the varied result of a complex mixture of cultures throughout history, and are separated by clear social, cultural, linguistic or geographic barriers. The stronger genetic differences between closely related populations occur in the northern third of Spain, a phenomenon commonly known as "micro-differentiation". It has been argued and discussed how this form of genetic structuring can be related to both the rugged landscape and the ancient societies of Northern Iberia, but this is difficult to test in most regions due to the intense human mobility of previous centuries. Nevertheless, the Spanish autonomous community of Asturias shows a complex history which hints of a certain isolation of its population. This, joined together with a difficult terrain full of deep valleys and steep mountains, makes it suitable for performing a study of genetic structure, based on mitochondrial DNA and Y-Chromosome markers. Our analyses do not only show that there are micro-differentiation patterns inside the Asturian territory, but that these patterns are strikingly similar between both uniparental markers. The inference of barriers to gene flow also indicates that Asturian populations from the coastal north and the mountainous south seem to be relatively isolated from the rest of the territory. These findings are discussed in light of historic and geographic data and, coupled with previous evidence, show that the origin of the current genetic patterning might indeed lie in Roman and Pre-Roman sociopolitical divisions.

Mitochondrial diversity patterns and the Magdalenian resettlement of Europe: new insights from the edge of the Franco-Cantabrian refuge.

Phylogeography of the mitochondrial lineages commonly found in Western Europe can be interpreted in the light of a postglacial resettlement of the continent. The center of this proposal lies in the Franco-Cantabrian glacial refuge, located in the northern Iberian Peninsula and Southwestern France. Recently, this interpretation has been confronted by the unexpected patterns of diversity found in some European haplogroups. To shed new lights on this issue, research on Iberian populations is crucial if events behind the actual genetics of the European continent are to be untangled. In this regard, the region of Asturias has not been extensively studied, despite its convoluted history with prolonged periods of isolation. As mitochondrial DNA is a kind of data that has been commonly used in human population genetics, we conducted a thorough regional study in which we collected buccal swabs from 429 individuals with confirmed Asturian ancestry. The joint analysis of these sequences with a large continent-wide database and previously published diversity patterns allowed us to discuss a new explanation for the population dynamics inside the Franco-Cantabrian area, based on range expansion theory. This approximation to previously contradictory findings has made them compatible with most proposals about the postglacial resettlement of Western Europe.

Tabaco de liar: una prioridad de salud pública y consumo / Fine-cut tobacco: a priority for public health and consumer advocacy

Objetivos Valorar si el tabaco de liar supone un problema en materia de salud pública y consumo. Métodos Se estudia el 70% del mercado español. En materia de salud pública, se analizan los contenidos de nicotina, alquitrán y monóxido de carbono del humo, y se comparan con los de los cigarrillos convencionales. En materia de consumo, se examina el etiquetado. Resultados Los contenidos de nicotina, alquitrán y monóxido de carbono alcanzan valores de hasta el 70%, el 85% y el 84%, respectivamente, más de lo permitido para los cigarrillos convencionales. El 67% de las muestras no indican los contenidos de nicotina y alquitrán, y el 100% el monóxido de carbono. A pesar de la existencia de etiquetado, éste no garantiza una información suficiente al consumidor. Conclusiones El tabaco de liar supone un problema tanto en materia de salud pública como en consumo. Por ello, sería necesaria alguna medida que permita resolver esta cuestión (AU)

Objectives To study whether fine-cut tobacco poses a problem for public health and consumer affairs. Methods We analyzed up to 70% of the fine-cut tobacco market in Spain. Regarding public health, the contents of nicotine, tar and carbon monoxide were analyzed and compared with levels in conventional cigarettes. Concerning consumer affairs, the labeling of samples was checked. Results The contents of nicotine, tar and carbon monoxide reached values of 70%, 85% and 84%, respectively. These values are higher than those allowed in conventional cigarettes. A total of 67% of the samples analyzed did not show nicotine and tar contents on the labeling. None of the labels showed carbon monoxide contents. The presence of labeling per se did not guarantee sufficient information for consumers. Conclusions Fine-cut tobacco is a problem in both public health and consumer affairs. Solutions are required to resolve both problems (AU)

Interspecific hybridization increased in congeneric flatfishes after the Prestige oil spill.

Marine species with relatively low migratory capacity are threatened by habitat alterations derived from human activities. In November 2002 the tanker Prestige sank off the Spanish northwest coast releasing 70,000 tons of fuel and damaging biota in the area. Despite efforts to clean the damaged areas, fuel remnants have affected marine species over the last nine years. This study is focused on two flatfish, Lepidorhombus boscii (four-spotted megrim) and L. whiffiagonis (megrim), whose spawning areas are located at the edge of the continental platform. We have analyzed megrim samples from North Spanish and French waters obtained before and after the oil spill. Genotypes at the nuclear marker 5S rDNA indicate a significant increase in interspecific hybridization after the Prestige accident, likely due to forced spawning overlap. The mitochondrial D-Loop region was employed for determining the direction of hybrid crosses, which were most frequently L. boscii female x L. whiffiagonis male. Reduced ability of L. boscii females to select conspecific mates would explain such asymmetric hybridization. To our knowledge this is the first time that increased hybridization between fish species can be associated to an oil spill. These results illustrate the potential long-term effect of petrol wastes on wild fish species.

[Fine-cut tobacco: a priority for public health and consumer advocacy]. / Tabaco de liar: una prioridad de salud pública y consumo.

OBJECTIVES: To study whether fine-cut tobacco poses a problem for public health and consumer affairs. METHODS: We analyzed up to 70% of the fine-cut tobacco market in Spain. Regarding public health, the contents of nicotine, tar and carbon monoxide were analyzed and compared with levels in conventional cigarettes. Concerning consumer affairs, the labeling of samples was checked. RESULTS: The contents of nicotine, tar and carbon monoxide reached values of 70%, 85% and 84%, respectively. These values are higher than those allowed in conventional cigarettes. A total of 67% of the samples analyzed did not show nicotine and tar contents on the labeling. None of the labels showed carbon monoxide contents. The presence of labeling per se did not guarantee sufficient information for consumers. CONCLUSIONS: Fine-cut tobacco is a problem in both public health and consumer affairs. Solutions are required to resolve both problems.

Introducing human population biology through an easy laboratory exercise on mitochondrial DNA.

This article describes an easy and cheap laboratory exercise for students to discover their own mitochondrial haplogroup. Students use buccal swabs to obtain mucosa cells as noninvasive tissue samples, extract DNA, and with a simple polymerase chain reaction-restriction fragment length polymorphism analysis they can obtain DNA fragments of different sizes that can be visualized in agarose gels. The analysis of these fragments can reveal the mitochondrial haplogroup of each student. The results of the exercise can be used to provide additional insights into the genetic variation of human populations.