The increasing availability of reference libraries of DNA barcodes (RLDB) offers the opportunity to the screen the level of consistency in DNA barcode data among libraries, in order to detect possible disagreements generated from taxonomic uncertainty or operational shortcomings. We propose a ranking system to attribute a confidence level to species identifications associated with DNA barcode records from a RLDB. Here we apply the proposed ranking system to a newly generated RLDB for marine fish of Portugal. Specimens (n = 659) representing 102 marine fish species were collected along the continental shelf of Portugal, morphologically identified and archived in a museum collection. Samples were sequenced at the barcode region of the cytochrome oxidase subunit I gene (COI-5P). Resultant DNA barcodes had average intra-specific and inter-specific Kimura-2-parameter distances (0.32% and 8.84%, respectively) within the range usually observed for marine fishes. All specimens were ranked in five different levels (A–E), according to the reliability of the match between their species identification and the respective diagnostic DNA barcodes. Grades A to E were attributed upon submission of individual specimen sequences to BOLD-IDS and inspection of the clustering pattern in the NJ tree generated. Overall, our study resulted in 73.5% of unambiguous species IDs (grade A), 7.8% taxonomically congruent barcode clusters within our dataset, but awaiting external confirmation (grade B), and 18.7% of species identifications with lower levels of reliability (grades C/E). We highlight the importance of implementing a system to rank barcode records in RLDB, in order to flag taxa in need of taxonomic revision, or reduce ambiguities of discordant data. With increasing DNA barcode records publicly available, this cross-validation system would provide a metric of relative accuracy of barcodes, while enabling the continuous revision and annotation required in taxonomic work.

The 3 lamprey species, sea lamprey Petromyzon marinus L., European river lamprey Lampetra fluviatilis L. and European brook lamprey L. planeri Bloch, that inhabit the Iberian Peninsula are of conservation concern. They are considered either Vulnerable, Critically Endangered, and even Extinct in different regions of this area mainly due to habitat loss and population fragmentation. Although several other factors contribute to the decline of lamprey populations in Iberian rivers, obstacles to migration (dams and weirs) are probably the most widespread and significant, causing an estimated 80% loss of accessible habitat in most river basins. We analysed historical records from all main Iberian rivers before the construction of impassable dams became widespread, and found that lampreys were consistently present in the upper reaches. The un - blocking of the lower stretches of major river basins and the restoration of former spawning sites and larval habitats should be considered as priority measures for the conservation of these species. Identification of Special Areas of Conservation to be included in the Natura 2000 European network can also be very relevant for lamprey conservation

MJ Alves, MM Coelho, MJ Collares-Pereira; Diversity in the Reproductive Modes of Females of the Rutilus alburnoides Complex (Teleostei, Cyprinidae): A Way to A