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Tiziana Lupo

  • Biodiversity Journal, 9 (2): 115-120

    Viviana Giangreco, Ignazio Sammarco, Tiziana Lupo, Sergio Migliore & Stefano Reale
    Molecular techniques employed to trace the Sicilian ovines
    https://doi.org/10.31396/Biodiv.Jour.2018.9.2.115.120

    ABSTRACT
    Genotyping strategies are aimed at defining the genetic profile of individuals through the identification of STRs sequences. The applied methodologies are able to ensure the traceability of the meat along the production chains and the control of the correct animal sampling on the farms. However, the discriminative capacity of alleles is studied through the establishment of the allelic frequency in the ovine population of the territory. This may depend on factors such as race, degree of inbreeding, and local selections. In the research of genetic identity in particular, it is exploited that the probability that two different individuals possess the same genetic pattern is equal to the frequency of that genotype in the population under examination and that the frequency of a genotype characterized by more loci is equal to the product of the frequencies of each single genotype (locus) observed. Therefore, we set the task of fixing and tabulating the data of the genetic profiles of the autochthonous breeds that can then be exploited for the traceability investigations of the animals, according to the application of specific algorithms. In practice, we aim to establish and create the starting point for the interpretation of all the genetic data obtained from the analysis of the Sicilian ovine population, whatever the application to do with it. The ultimate goal of this work is the elaboration of allelic panels typical of the sheep populations that represent the starting point for all genetic tests of forensic investigations. In fact, the discovery of particular alleles identify the tabulated frequency representing the genetic variability distribuited in the region. This has the effect of minimizing the identification errors that are spread in the animal population. We can state that from the analysis of allele frequencies developed by Genalex we can obtain expected heterozygosity data according to Hardy-Weinberg law and the obtained heterozygosity data typical for native breeds. All the allele frequencies were employed to create a database containing all the genotypes. These data were useful in the forensic field for the attribution of the kinships in the sheep.

  • Biodiversity Journal, 9 (2): 107-114

    Viviana Giangreco, Claudio Provito, Luca Sineo, Tiziana Lupo, Floriana Bonanno & Stefano Reale
    Identification of subspecies and parentage relationship by means of DNA fingerprinting in two exemplary of Pan troglodytes (Blumenbach, 1775) (Mammalia Hominidae)
    https://doi.org/10.31396/Biodiv.Jour.2018.9.2.107.114

    ABSTRACT
    Four chimpanzee subspecies (Mammalia Hominidae) are commonly recognised: the Western Chimpanzee, P. troglodytes verus (Schwarz, 1934), the Nigeria-Cameroon Chimpanzee, P. troglodytes ellioti, the Central Chimpanzee, P. troglodytes troglodytes (Blumenbach, 1799), and the Eastern Chimpanzee, P. troglodytes schweinfurthii (Giglioli, 1872). Recent studies on mitochondrial DNA show the incorporation of P. troglodytes schweinfurthii in P. troglodytes troglodytes, suggesting the existence of only two subspecies: P. troglodytes troglodytes in Central and Eastern Africa and P. troglodytes verus; P. troglodytes ellioti in West Africa. The aim of the present study is twofold: first, to identify the correct subspecies of two chimpanzee samples collected in a Biopark structure in Carini (Sicily, Italy), and second, to verify whether there was a kinship relationship between the two samples through techniques such as DNA barcoding and microsatellite analysis. DNA was extracted from apes’ buccal swabs, the cytochrome oxidase subunit 1 (COI) gene was amplified using universal primers, then purified and injected into capillary electrophoresis Genetic Analyzer ABI 3130 for sequencing. The sequence was searched on the NCBI Blast database. In addiction, the microsatellite analysis was performed on the same machine for parentage detection among samples, and data were analyzed with GenMapper software. Our results show that both samples were P. troglodytes troglodytes, while the analysis of the microsatellite results in an unclear relationship between two chimpanzee samples.

  • Biodiversity Journal, 10 (4): 457-461 - MONOGRAPH

    Maria Flaminia Persichetti, Viviana Giangreco, Antonio Gentile, Tiziana Lupo, Gabriele Ciaccio & Santo Caracappa
    Molecular barcoding applied to the Mediterranean turtles biological matrices (Reptilia Cheloniidae)
    https://doi.org/10.31396/Biodiv.Jour.2019.10.4.457.461

    ABSTRACT
    Chelonia mydas (Linnaeus, 1758) together with Caretta caretta (Linnaeus, 1758) is the most representative Cheloniidae species in the Mediterranean basin. Currently, at the National Reference Centre in the “Istituto Zooprofilattico” of Sicily (Italy), damaged subjects are rehabilitated before they are released again. Clinical, physiological and molecular parameters were collected from each subject. We analysed 46 turtles which samples were collected. Species specific Cytochrome oxidase I sequences for the identification of marine turtle species were obtained. Barcoding is a new tool of classical taxonomy that allows the characterisation of living species and the differentiation of very morphologically similar species. It is a practical tool that can be used in cases of damaged samples and is also useful for taxonomical characterisation of specimen at immature development stages. In our region, in the centre of the Mediterranean area, we represent a reference centre for injured animals both stranded on the beach and captured in offshore.Turtles caught in fishing lines generally retain the fishing hooks in their throat or oesophagus, as visible by X-ray investigations. After the cure and samples collection, the animals are released into the sea. The polymorphisms could be related to the geographical distance of the turtles following different routes during their life. The large-scale sequencing of a single or few genes in taxonomic studies, denominated by species barcoding, aims at offering a practical method for species identification, as well as for providing insights into the evolutionary diversification of life.

  • Biodiversity Journal, 10 (4): 445-450 - MONOGRAPH

    Viviana Giangreco, Tiziana Lupo, Ignazio Sammarco, Maurizio Bivona, Gabriele Ciaccio, Luca Sineo & Stefano Reale
    Genetic database development for the characterisation of Sicilian sheep population
    https://doi.org/10.31396/Biodiv.Jour.2019.10.4.445.450

    ABSTRACT
    The most representative sheep in Sicily are Belicina, Comisana, Pinzirita, Barbaresca and the crossbred derived sheep from all this species. In this study, the allelic frequencies of the Sicilian sheep population were investigated. It currently represents the best way to determine the genetic identity and/or family even with limited amounts of sample or when the DNA is degraded. The aim of the study was to provide a reference data bank and to evaluate a microsatellite panel for pedigree analysis as suggested by the International Society for Animal Genetics (ISAG). There are various studies on European sheep, but few datasets were developed on the population of Sicilian sheep. The reference database will include allele frequencies at each locus and will determine genetic parameters for Sicilian ovine species selection. Our results indicated that Hardy Weinberg equilibrium was not always maintained. These results could be explained by a non-random mating. The database is useful to investigate the relationship, the parentage the meat traceability and in disease control programs. The standardized panels of allele frequencies represent a molecular fingerprinting characterizing the subjects with very high definition level and can be useful to control all the livestock. The parentage identification could be important for the veterinary police to investigate the theft or the animal substitutions in the Sicilian farms.