The Etruscan chub Squalius lucumonis (Bianco, 1983) is an endemic endangered fish inhabiting the lakes and rivers of Central Italy. A microsatellite-enriched genomic library was constructed in order to develop microsatellite loci and eight polymorphic loci were isolated from this species to assess the level of genetic diversity in fifty individuals of S. lucumonis collected in the Umbria Region. The number of alleles ranged from four (Sluc12) to 18 (Sluc4). Observed and expected heterozygosity per locus ranged from 0.05 (Sluc12) to 0.80 (Sluc4; Sluc7 and Sluc11) and from 0.22 (Sluc12) to 0.90 (Sluc4), respectively. Furthermore, these loci were tested on ten other Cyprinidae species and four freshwater fish species belonging to other families. Successful cross-priming amplifications were obtained for several species of Cyprinidae, whereas no positive results were obtained for other species. The availability of the reported microsatellite loci will facilitate population structure investigations of these species aiming to phylogeographical approaches and conservation strategies.
Development perturbations may affect the regular phenotype and are commonly measured through fluctuat-ing asymmetry (FA) levels. Short-term climatic variations, that modify the temperature, can influence chemical and physical water characteristics. Fishes have been used as model organisms for studying stress-induced changes in body symmetry, since they are ectothermic, good bioindicators, easy to find and having economic relevance. Northern pike being a holoarctic, big, edible, top predator is one of the most economically important freshwater fish for recreational and commercial fisheries and freshwater ecosystems management. The isolated population of Lake Trasimeno (Central Italy)—in good health conditions and that can be considered one of the genetically best conserved of Italy—, was chosen as model. FA, seven mi-crosatellite loci and early developmental stages were investigated in order to correlate the developmental stability of this population to its genetic variability and to environmental perturbations. The results obtained underlined a positive correlation (>>0.40) between FA indexes and temperature; the non-parametric Kruskall- Wallis test showed significant differences in FA levels for some FA indexes and parameters. Over-all results underlined that FA increased in individuals grown at a temperature above 8°C as compared with those grown at 5°C or at lower temperatures. Both positive and negative correlations between FA parameters and chemical and physical water characteristics were shown. The comparison of genetic and FA data under-lined a low correlation between microsatellite and FA pairwise distances, nevertheless a positive and signifi-cant correlation emerged for some FA measurements and microsatellite data. In particular, only Elu87 locus showed a statistical significant correlation versus total FA. Finally, as expected, results indicated that the in-cubation time was temperature-dependent; the ODT was in the range 8–10°C and lower and higher tempera-tures caused drastic embryo mortality. These results showed robust correlations, both positive and negative, between some FA parameters and chemical and physical characters and were in agreement with the assump-tion that temperature variations as well as pH, conductibility and chloride variation, may increase molecular perturbations and, subsequently, the global developmental noise during development. These data suggest that FA could be considered a measure of animal welfare. The relative breeding easiness of this species may be a valid tool for the estimation of controlled environmental stress influences, not only of thermal origin, and a valid information basis for studies on wild populations. Furthermore, it has long been debated whether FA levels depend upon genetic variability, the particular molecular marker notwithstanding, and whether it is possible to use one or more molecular markers to better understand FA data. The Mantel tests performed in this study showed very interesting correlations between FA and the investigated microsatellites. For the lack of a linkage map for the investigated microsatellite loci, it is presently impossible to establish the relation-ships between the FA parameter and the microsatellite loci that the Mantel test defines as correlated.
Grey partridge and Red-legged partridge are Galliformes needing special conservation strategies. Reintroduction may represent conservation solutions solely with the support of an in-depth genetic and ecologic evaluation, particularly of grey partridge, of which an Italian subspecies was described. Protocols for conservative DNA isolation are fundamental to study breeders and wild samples. For these reasons, two DNA extraction protocols on different kinds of feathers (P1–P6), oral and anal swabs were tested. DNA suitability was assayed through PCR amplification and capillary sequencing of both 408 bp of Control Region and 658 bp of COI. Both protocols yielded well-amplifiable DNA, they were not time and money consuming and did not require harmful reagents. Sequences of DNA extracted through Wizard Genomic DNA Purification Kit (WGDPK) were better than those based on polyvinylpyrrolidone (PVP); buccal swab, P3 and P5 seemed the best sampling methods, in particular, P5-feather sampling did not affect the flying capacity.