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Summary Accurate quantitative assays are required for enforcing food labelling procedures and preventing food ingredient contamination, misdescription and fraud. Two species-specific TaqMan systems and a eukaryotic endogenous TaqMan system were combined in a real time PCR approach to achieve quantification of chamois and pyrenean ibex meats in meat mixtures. The caprinae-specific systems were based on the amplification of a 133 bp fragment of the 12S rRNA gene from chamois DNA, whereas an 88 bp fragment from the D-loop region was amplified from pyrenean ibex DNA. Universal primers amplified a 141 bp fragment on the nuclear 18S rRNA gene from eukaryotic DNA. Analysis of experimental raw and heat treated binary mixtures of chamois and pyrenean ibex meat in a swine meat matrix demonstrated the suitability of the assay for the detection and quantification of the target DNAs in the range of 0.1–25%, depending on the species and treatment of the meat samples.

A real-time quantitative polymerase chain reaction (PCR) technique was developed for the quantification of chamois and pyrenean ibex DNAs in meat mixtures by using a SYBR green detection platform. Two species-specific systems and a eukaryotic endogenous system were combined in the real-time PCR approach to quantify the target species. In the specific systems, a 133 base pair (bp) fragment of the 12S rRNA gene was amplified from chamois DNA, and an 88 bp fragment from the D-loop region was amplified from pyrenean ibex DNA. In the endogenous system, universal primers amplified a 141 bp fragment on the nuclear 18S rRNA gene from eukaryotic DNA. The threshold cycle values obtained with the 18S rRNA primers were used to normalize those obtained from chamois- or pyrenean ibex-specific systems, serving as endogenous control for the total content of PCR-amplifiable DNA in the sample. Analysis of experimental raw and heat-treated binary mixtures of chamois and pyrenean ibex meat in a swine meat matrix demonstrated the suitability of the assay for the detection and quantification of the target DNAs in the range of 0.10.8, depending on the species and treatment of the meat samples.

A polymerase chain reaction (PCR) assay was developed for the identification of meats from chamois (Rupicapra rupicapra), pyrenean ibex (Capra pyrenaica), and mouflon (Ovis ammon) by using oligonucleotides targeting mitochondrial D-loop sequences. A D-loop region (∼700-1000 bp) was firstly amplified and sequenced from various game and domestic meat DNAs, and three primer sets were then designed on the basis of nucleotide multialignment of the generated D-loop sequences. As expected from sequence analysis, PCR amplification of the targeted D-loop fragments was successfully achieved from chamois (88 bp), pyrenean ibex (178 bp), and mouflon (155 bp) meats, showing adequate specificity and reproducibility against a number of game and domestic meats. Mouflon and sheep meats were amplified together in accordance to the high nucleotide identity of their mt D-loop sequences. In this work, satisfactory amplification was also accomplished in the analysis of experimentally pasteurized (72°C for 30min) and sterilized (121°C for 20min) meats, with a detection limit of ∼0.1% for each of the targeted species. The proposed PCR assay represents a rapid and straightforward method for the detection of possible adulterations in game meat products.

I once found myself gazing at a forlorn exhibit: a nondescript stuffed pigeon – the remains of Martha, a passenger pigeon and the last member of a once hyperabundant species rendered extinct with her death in 1914. I am reminded of other last survivors, such as Lonesome George the giant tortoise, Richard Henry Kakapo, and Celia the Pyrenean ibex. Two things occur to me. First, if you are a member of an endangered species and someone names you, watch out; going on past records, your kind's days are numbered.

The Pyrenean ibex, an impressive mountain goat that lived in the central Pyrenees in Spain, went extinct in 2000. But a group of scientists is trying to revive the species by cloning cells from its last living member. A previous attempt in 2003 failed, but renewed interest in "de-extinction" in general and a private donation have allowed the scientists to try again.

El avance de las ciencias biomédicas está llegando a campos inexplorados y sorprendentes; en fecha reciente, se ponderó la posibilidad de recuperar especies extintas al publicarse la resurrección de la primera subespecie extinta: el bucardo. No obstante, la incorporación de técnicas reproductivas avanzadas en el mundo de la conservación biológica se mantiene hasta ahora limitada e incluso, en ocasiones, mal encaminada. Los factores involucrados en este pobre desarrollo son diversos, pero uno de los más importantes es el rechazo de la comunidad conservacionista a las estrategias de alta tecnología. Sin embargo, considerando que México es un país megadiverso, con un gran número de especies de mamíferos amenazados, es urgente la necesidad de que se incorporen herramientas biotecnológicas al estudio y conservación de las especies del país, y más aún, que sean las mismas comunidades científicas latinoamericanas quienes desarrollen y utilicen sus propias herramientas. En esta nota, se discuten los avances científicos que proponen la transferencia somática nuclear como una herramienta de conservación, sus posibilidades reales, las necesidades y sugerencias para su aplicación y desarrollo con el fin de convertir la biotecnología moderna en instrumento para la lucha por la conservación de las especies amenazadas de México y Latinoamérica.

Recent accomplishments in the fields of nuclear transfer and genomics, such as the cloned offspring production from frozen mouse cells, cryopreserved at not too low temperatures without cryoprotectors; or the sequencing of wooly mammoth genome, have opened the opportunity for the revival of extinct species. As expected, they are receiving a lot of publicity in the media and also scientific attention. Furthermore, it was recently published the "revival" of the first extinct subspecie: the Pyrenean ibex (Capra pyrenaica pyrenaica), a wild goat extinct in 2000. This strengthens the field of cloning as it had been tarnished by induced pluripotent stem cells (iPS) and other methods of reprogramming. However, for biological conservation purposes, cloning is not generally accepted as an alternative for animal conservation, and there is an ongoing debate between reproductive scientists and conservation specialists. Although we believe that nuclear transfer technologies have an opportunity in conservation efforts for some species that are on the brink of extinction and that population status, geographical isolation, reproductive characteristics, and human pressure create a situation that is almost unsustainable. In this article we discuss the barriers in cloning mammoths and cloning controversies in conservation from a zoological perspective, citing the species that might benefit from nuclear transfer techniques in the arduous journey so as not to disappear forever from this, our world.

[EN]: During the last century, human activities have promoted large changes in habitats, leading to changes in the abundance and composition of mammal communities. Some species have been favoured intentionally (specially those with high economic importance), or unintentionally (i.e. opportunistic species), introduced las some livestock species), or persecuted (i.e. species that affect livestock or game). In this paper, we show and explain the changes in the composition of the large mammal community in Aragon, a northeastern Spanish area where there have been extinctions (lynx, Lynx sp.); introductions (moufflon, Ovis ammon; fallow deer, Dama dame); reintroductions (red deer, Cervus elaphus); and particularly natural expansions (wild boar, Sus scrofa; roe deer, Capreolus capreolus; red deer; Pyrenean chamois, Rupicapra pyrenaica pyrenaica). Some taxa are currently almost extinct (brown bear, Ursus arctos; Pyrenean ibex, Capra pyrenaica pyrenaica) and others are recovering after getting close to extinction (wolf, Canis lupus). Three interconnected causes may explain the clear increment of ungulates in our study area: (1) the abandonment of rural activities, (2) the subsequent decrease of livestock, and (3) the increase of forest habitats. None of those species are currently below 150 % of their range in the 19(th) century. The setting up of hunting reservations and the design of a hunting legislation have also contributed to the observed changes. Traditionally, large predators have not been favoured by human activities, while compensatory protection measures carried out have been scarce or came too late. While some of these species are probably extinct in the area, the remaining occupy currently less than one fourth of their former range and need urgent conservation measures for to recover., [FR]: Au cours du siecle dernier, les activites humaines ont provoque de grands changements dans les habitats, ce qui a conduit ä des modifications de l'abondance et de la composition des communautes de mammiferes. Certaines especes ont ete favorisees intentionnellement (en particulier celles qui ont une grande importance economique), ou involontairement (especes opportunistes), introduces (certaines especes de betail), ou au contraire consideres comme nuisibles au betail ou aux animaux de chasse. Nous montrons ici les modifications dans la composition de Pensemble de la faune des grands mammiferes en Aragon, region du nord-est de PEspagne ou il y a eu des extinctions (lynx), des introductions (mouflon, daim), des reintroductions (cerf), et aussi des expansions naturelles (sanglier, chevreuil, cerf, isard). Certaines especes sont preque eteintes (ours, ibex), et d'autres se retablissent apres avoir ete proches de l'extincition (loup). Trois causes liees entre elles peuvent expliquer Faccroissement des ongules dans cette region: 1) abandon des activites rurales; 2) la reduction du betail; 3) Paccroissement des habitats de foret. Ces especes sont generalement bien en dessous de leur repartition au XIXC siecle. L'etablissement de reserves de chasse et d'une legislation particuliere ont aussi contribue aux changements constates. Traditionnellement, les grands predateurs n'ont pas ete favorises par les activites humaines et les mesures de protection sont arrivees trop tard. Certaines especes sont eteintes et d'autres n'occupent plus que le quart de leur repartition primitive. II est done urgent de prendre des mesures de conservation., Most of the data have been extracted from several reports supported by the Aragon Government Fish and Game Services.

International audience; Cementochronology of Pyrenean Ibex (Capra Hircus Pyrenaica) teeth from Les Eglises Cave (Ussat, Ariège). Cementum increment analysis is considered a very useful method to study the seasonality of Paleolithic sites. The method is applicable to a wide range of taxa but it needs to be validated for each one of them. In this paper, we analyse remains of Pyrenean Ibex (Capra hircus pyrenaica) coming from Les Eglises cave (Ussat-Ariège-France) We compare the seasonality results established by cementum increment analysis with those independently established by another author on the basis of eruption-wear patterns. Image analysis software has been used for cementum annulation interpretations.; La cémento-chronologie est reconnue comme une méthode de première utilité pour déterminer la saisonnalité des habitats de chasseurs paléolithiques. Intéressant de nombreux groupes zoologiques, cette méthode demande à être validée pour chacun d'eux. Ici, les auteurs s'intéressent au Bouquetin des Pyrénées et comparent les résultats qu'ils ont collectés par la méthode de la cémento-chronologie à d'autres obtenus à partir de l'étude du degré d'évolution dentaire. Ils illustrent également l'aide apportée, pour la lecture, par un logiciel de traitement et d'analyse d'image.

Cementochronology of Pyrenean Ibex (Capra Hircus Pyrenaica) teeth from Les Eglises Cave (Ussat, Ariège). Cementum increment analysis is considered a very useful method to study the seasonality of Paleolithic sites. The method is applicable to a wide range of taxa but it needs to be validated for each one of them. In this paper, we analyse remains of Pyrenean Ibex (Capra hircus pyrenaica) coming from Les Eglises cave (Ussat - Ariège - France) We compare the seasonality results established by cementum increment analysis with those independently established by another author on the basis of eruption-wear patterns. Image analysis software has been used for cementum annulation interpretations., La cémento-chronologie est reconnue comme une méthode de première utilité pour déterminer la saisonnalité des habitats de chasseurs paléolithiques. Intéressant de nombreux groupes zoologiques, cette méthode demande à être validée pour chacun d'eux. Ici, les auteurs s'intéressent au Bouquetin des Pyrénées et comparent les résultats qu'ils ont collectés par la méthode de la cémento-chronologie à d'autres obtenus à partir de l'étude du degré d'évolution dentaire. Ils illustrent également l'aide apportée, pour la lecture, par un logiciel de traitement et d'analyse d'image., Griggo Christophe, Pubert Eric. Cémento-chronologie de quelques dents de bouquetin des Pyrénées (Capra Hircus Pyrenaica) provenant de la Grotte des Eglises (Ussat, Ariège)/Cementochronology of Pyrenean Ibex (Capra Hircus Pyrenaica) teeth from Les Eglises Cave (Ussat, Ariège). In: Paléo, n°11,1999. pp. 177-185.

International audience; Cementochronology of Pyrenean Ibex (Capra Hircus Pyrenaica) teeth from Les Eglises Cave (Ussat, Ariège). Cementum increment analysis is considered a very useful method to study the seasonality of Paleolithic sites. The method is applicable to a wide range of taxa but it needs to be validated for each one of them. In this paper, we analyse remains of Pyrenean Ibex (Capra hircus pyrenaica) coming from Les Eglises cave (Ussat-Ariège-France) We compare the seasonality results established by cementum increment analysis with those independently established by another author on the basis of eruption-wear patterns. Image analysis software has been used for cementum annulation interpretations.; La cémento-chronologie est reconnue comme une méthode de première utilité pour déterminer la saisonnalité des habitats de chasseurs paléolithiques. Intéressant de nombreux groupes zoologiques, cette méthode demande à être validée pour chacun d'eux. Ici, les auteurs s'intéressent au Bouquetin des Pyrénées et comparent les résultats qu'ils ont collectés par la méthode de la cémento-chronologie à d'autres obtenus à partir de l'étude du degré d'évolution dentaire. Ils illustrent également l'aide apportée, pour la lecture, par un logiciel de traitement et d'analyse d'image.

The prevention of fraudulent labeling of game meat constitutes an important part of food regulatory control and quality assurance systems. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis based on mitochondrial deoxyribonucleic acid (DNA) was developed for authentication of meats from chamois (Rupicapra rupicapra), pyrenean ibex (Capra pyrenaica), and mouflon (Ovis ammon). Amplification and restriction site analysis of a DNA fragment about 720 base pairs (bp) from the mitochondrial 12S rRNA gene of all analyzed species permitted the selection of MseI and ApoI endonucleases for meat speciation. The 12S rRNA restriction profiles obtained allowed the unequivocal identification of chamois, pyrenean ibex, and mouflon/sheep and their differentiation from meats of domestic species such as cattle, goat, and swine. The highly variable mitochondrial D-loop gene was also targeted to attempt discrimination between mouflon and sheep meats. A D-loop region (700-1000 bp) was amplified and sequenced in all game and domestic species analyzed, and a primer set was designed for the selective amplification of a 370 bp DNA fragment from mouflon and sheep. PCR-RFLP analysis with the selected MaeII enzyme generated a single electrophoretic profile characteristic for sheep, whereas 3 different fragment patterns were obtained for mouflon meats. Consequently, the PCR-RFLP technique developed can be routinely applied in inspection programs in order to verify the correct labeling of game species.