European plaice (Pleuronectes platessa) is quite frequently substituted with other flatfish, especially with Yellow fin sole (Limanda aspera), which differs not only in meat quality but also ...particularly in its origin and manipulation chain. We propose an integrated approach of laboratory and in silico mtDNA PCR-RFLP procedures generating a set of restriction patterns easily resolvable in agarose gel, able to discriminate with certainty P. platessa from other 20 flatfish species. The herein proposed procedure is an economical and valid tool in detecting mislabelled seafoods.
The brook chub, Squalius lucumonis (Bianco, 1983), is an endemic endangered species inhabiting the rivers of Central Italy. Being subject to hybridizations with other cyprinids, particularly the ...congeneric S. squalus (Bonaparte, 1837), genetic overestimation can emerge when molecular markers are applied and mtDNA barcoding is not sufficient to ensure a correct attribution, due to its maternal inheritance. In this perspective, a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) approach on rDNA is proposed to rapidly discriminate S. lucumonis from nine other cyprinids. For its capacity to identify hybrids of these species with S. lucumonis, this method should be considered as a fundamental tool to be applied prior to the application of other molecular markers in extensive genetic characterizations for phylogenetic, phylogeographic, population and management analyses.
Genetic analyses allowed management and conservative programmes specifically designed for biodiversity conservation purposes. To date, no information is available for the
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e definition of any ...Italian northern pike population, because of their wide, highly fragmented, distribution. Low gene flow levels may have caused some of the health population problems emerging from professional fisheries data, underlining a marked reduction of these populations. This paper reports the first estimate of genetic
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e of the Lake Trasimeno's northern pike, by means of microsatellites data, over the last four decades compared with five other significant Italian populations.
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e values were evaluated through specific software and
θ estimator. The
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e results confirm that, despite a marked reduction between 1966 and 1997 and a less than optimal status, the Trasimeno population shows a positive trend in its health status, presumably due to proper management programmes. Considerations on the biological conservation and management of this pike population are also reported.
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.
The two freshwater sponges Ephydatia fluviatilis and Ephydatia mülleri belong to the widespread Spongillidae family. Their morphological tracts are very similar and can be distinguished mainly on the ...basis of their gemmuloscleres. However, when gemmules are absent it is essential to have an unambiguous species attribution for a population genetic study based on fresh tissues and historical collections. This article reports a simple Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis, applied to DNA extracted from both gemmules and fresh tissues in order to discriminate between the two congeneric E. fluviatilis and E. mülleri. Such a biomolecular method is based on the discriminative enzymes' digestion of each of the three amplified fragments 5.8S-ITS2-28S, D3 domain of the 28S subunit and COI. Two restriction enzymes were tested for a 620-642 bp fragment of 5.8S-ITS2-28S and for a 342 bp fragment of the D3 domain of the 28S, one restriction enzyme was tested for a 681 bp fragment codifying for COI. Obtained digestion patterns were diagnostic for each of the two species, providing a relatively simple, fast and cheap method for species attribution compared to sequencing.
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.
The two freshwater sponges Ephydatia fluviatilis and Ephydatia muelleri belong to the widespread Spongillidae family. Their morphological tracts are very similar and can be distinguished mainly on ...the basis of their gemmuloscleres. However, when gemmules are absent it is essential to have an unambiguous species attribution for a population genetic study based on fresh tissues and historical collections. This article reports a simple Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis, applied to DNA extracted from both gemmules and fresh tissues in order to discriminate between the two congeneric E. fluviatilis and E. muelleri. Such a biomolecular method is based on the discriminative enzymes' digestion of each of the three amplified fragments 5.8S-ITS2-28S, D3 domain of the 28S subunit and COI. Two restriction enzymes were tested for a 620-642 bp fragment of 5.8S-ITS2-28S and for a 342 bp fragment of the D3 domain of the 28S, one restriction enzyme was tested for a 681 bp fragment codifying for COI. Obtained digestion patterns were diagnostic for each of the two species, providing a relatively simple, fast and cheap method for species attribution compared to sequencing.
Genetic diversity was analysed in a putative autochthonous brown trout (Salmo trutta fario L.) population (Monterivoso Stream, Tyrrhenian Apennine Slope) by means of seven microsatellite loci and ...PCR-RFLP of two mitochondrial (ND1 and ND5/6) and one nuclear (LDH-C1*) locus. Monterivoso data were compared to those obtained analysing populations of the same basin (Nera River) and of the Po basin; Irish populations were used as a source of Atlantic strain brown trout. Haplotypes distributions, heterozygosity, F-statistic and UPGMA analyses indicated a genetic diversification between these populations, suggesting a widespread alteration of the genetic structure due to repeated stocking with allochthonous material, mainly of Atlantic origin, that has partially polluted the Monterivoso population. This population appeared to be constituted of Mediterranean strain samples and might represent a residue of an indigenous pool: it shows high specificity characteristics and it is genetically separate from the others observed and from populations of the Adriatic slope of the Apennines. This population should be employed in managing and breeding programmes finalised to an eco-sustainable restocking of brown trout in the Nera River Basin, with periodic monitoring by genetic analyses.
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