$.'$) %*/,)' !!$$'*/,)'*! .# )$('$ ) ) ,* /.$*)--*$.$*)  *!

       )$('

111-+%*/,)'$.

-$ )

.'$)*/,)' *!)$(' $ ) 0*'/(  -/++' ( ).

 .# *)", -- , (*)/) 

**&*!-.,.- / -. $.*,$*(*$,'* 19 th Congress of Animal Science and Production Association ANIMAL BREEDING AND GENETICS

didymes of slaughtered animals can be a cheap alternative for pre- 6W serving male gametes. The aim of this study was to optimize the Demographic and genetic parameters in Miniature recovery of bull epididymal spermatozoa comparing the effects of Pinscher ( Canis familiaris ) Italian population two methods of extraction in combination with egg yolk extender. Testis from 23 Limousine bulls, ranging in age from 13 to 20 1 1 2 Stefano Paolo Marelli , Elena Colombo , Rita Rizzi , months, were collected at the abattoir. Spermatozoa were extract- 1 2 Michele Polli , Emanuela Tullo ed from the epididymes using two methods: the float-up and the 1Dipartimento di Scienze Animali; 2Dipartimento di Scienze retrograde flushing. The average time for extraction was 16 min- Veterinarie per la Sicurezza Alimentare, Università di Milano, utes in both methods. Within each methods half of testis were processed with an egg yolk extender and the second half with an Corresponding author: [email protected] egg-yolk free extender. Sperm concentration, motility, viability and morphology were evaluated. Statistical analyses were carried out The aim of this work was to investigate the genetic and demo- using the GLM procedure (SAS package v9.1). Methods of extrac- graphic parameters of the Italian population of Miniature tion, in combination with the presence of egg yolk in the extender, Pinscher (MP) and its evolution across generations to supply were considered as fixed effects; age of the bull and time for effective and objective tools for breed conservation and selection extraction as covariates. Sperm concentration was not significant- strategies. MP is a worldwide known toy breed of German origin, ly different using either the float-up method or the retrograde part of FCI group 2: Pinscher and Schnauzer - Molossoid Breeds - flushing (283.5±24.7 and 301.5±30.3 ×10 6 cells/mL, respectively). Swiss Mountain and Cattle Dogs. Height at withers ranges for Flushing technique was better than the float-up method in terms dogs and bitches between 25 to 30 cm and the weight between 4 of sperm quality, considering total motility (78.4±3.1% vs and 6 kg. The coat is short, dense and smooth, the colour is black 61.8±2,6%, P=0.0003; respectively) and viability (88.7±2.1% vs and tan or red-reddish- dark red brown; MP are wonderful compan- 77.6±1.7%, P=0.0003; respectively). Egg yolk influenced positively ion and guard dogs particularly adapted to apartment life style. motility and morphology in the float-up method, whereas Italian Kennel Club (ENCI) Stud Book official data of 25,065 sub- decreased viability in flushed samples. In conclusion, samples jects were analysed (12,880 females, 12,185 males); dates of birth extracted by flushing method, with egg yolk, showed a better ranged from 1965 to 2009. All the calculations were performed sperm quality. These data suggest the possible use of epididymal using ENDOG V 4.6 software. Mean maximum number of traced sperm collected by the flushing technique for the creation of cattle generations was 6.84, mean complete generations were 2.85 and semen cryobanks. mean equivalent generations 4.14. Average inbreeding coefficient (F) was 0.08 and average relatedness coefficient (AR) was 0.02. The maximum number of puppies entered was recorded in 2003 with 1111 new born MP. Alternating F values were calculated with 8W a constant increasing between the end of the seventies and the Genetic diversity and structure of Italian sheep first years of nineties (0.11, 1989), maximum calculated individual breeds inferred by genome-wide SNP analysis: F value was 0.55. MP are characterized by a significant number of overview of the “Italian Project for Sheep entries per year with low average inbreeding values, the coexis- Biodiversity” tence of two subpopulation: ‘show’ and ‘pet’ could be supposed. Further investigation are needed to better understand the genetic Fabio Pilla 1, Elena Ciani 2, Francesca Maria Sarti 3, Bianca Moioli 4, asset of MP breed. Antonello Carta 5, Donato Matassino 6, Paola Crepaldi 7, Roberta Ciampolini 8, Donata Marletta 9 Pier Luigi Acutis 10 , Nicolò P.P. Macciotta 11 , Paolo Ajmone Marsan 12 , Baldassarre 7W Portolano 13 1Dipartimento di Scienze Animali Vegetali e dell'Ambiente, Optimisation of bull epididymal spermatozoa extrac- Università del Molise, Italy tion techniques: a tool for improving gene banking 2Dipartimento di Fisiologia generale ed Ambientale, Università di Federica Turri 1,2 , Manuela Madeddu 1, Teresa Maria Gliozzi 1, Bari, Italy Gustavo Gandini 2, Flavia Pizzi 1 3Dipartimento di Biologia applicata, Università di Perugia, Italy 4 1CNR, Istituto di Biologia e Biotecnologia Agraria, Milano, Italy CRA, Centro di Ricerca per la Produzione delle Carni e il 2 Miglioramento Genetico, Monterotondo, Italy Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza 5 Alimentare, Università di Milano, Italy Agris Sardegna, Olmedo, Italy 6 Corresponding author: [email protected] Consorzio per la Sperimentazione Divulgazione e Applicazione di Biotecniche Innovative, Benevento, Italy Animal gene banks development is limited by high costs of cre- 7Dipartimento di Scienze Animali, Università di Milano, Italy ation. The recovery and freezing of viable sperm from the epi- 8Dipartimento di Patologia Animale, Profilassi e Igiene degli

[Ital J Anim Sci vol.10:s1, 2011] [page 69] Italian Journal of Animal Science 2011; volume 10: supplement 1 ANIMAL BREEDING AND GENETICS

Alimenti, Università di Pisa, Italy Bari, Italy 9Dipartimento di Scienze delle Produzioni Agrarie, Università di 4Dipartimento Patologia Animale, Profilassi ed Igiene degli ali- Catania, Italy menti, Università di Pisa, Italy 11 Dipartimento di Scienze Zootecniche, Università di Sassari, Italy 5Dipartimento Scienze Animali, Vegetali e dell’Ambiente, 12 Istituto di Zootecnica, Università Cattolica del Sacro Cuore, Università del Molise, Campobasso, Italy Piacenza, Italy Corresponding author: [email protected] 13 Dipartimento Demetra, Università di Palermo, Italy Corresponding author: [email protected] The proteomic analysis defines the identity, the structure and the relative abundance of proteins in a given type of cell and in a spe- So far, genetic diversity within and among Italian sheep breeds cific set of conditions. has been investigated almost exclusively by microsatellite markers The analysis of protein profile was carried out to identify possible and never at national scale. Recently, high density Single Nucleo - expression differences of muscular proteins between sheep tide Polymorphism (SNP) arrays have been developed for the breeds and use these peculiar profiles as trace back biomarkers. major livestock species, including sheep. In the second half of The analyses were carried out on a total of 98 samples of longis- 2010, a large national collaborative project was launched, within simus dorsi and of leg muscles pools from 49 ovine belonging to the context of the International Sheep Genome Consortium five breeds: (N=9), (N=10), Gentile di (ISGC), aiming at characterizing at the genomic level the main Puglia (N=10), Laticauda (N=10) and (N=10). The pro- Italian sheep breeds, together with some endangered local breeds. tein profile was investigated using analytical proteomic proce- Around 500 animals evenly distributed among 19 Italian breeds dures: 2DGE and MALDI-TOF/MS. The two-dimensional maps were (, Altamurana, , Bagnolese, Bergamasca, compared by image analysis using Image Master 2D-Platinum to , , , , , define the position and relative intensity (volume %) of every sin- Laticauda, Leccese, Massese, Pecora , , gle spot. In the range of our observation, image analysis highlight- , Sarda, , Valle del Belice) have been ed 15 spots common to all samples, identified as: Adenilate kinase, genotyped using a 50K SNP array. Molecular data have been DJ-1, Enolase (2 spot), Hemoglobin (Hb), Myoglobin (Myb) (2 pruned according to SNP minor allele frequency (MAF<0.01), fre- spot), Phosphatidilethanolamine binding protein (PEBP), quency of missing genotypes (Geno>0.1) and linkage disequilibri- Superoxide dismutase, Triosephosphate isomerase (TPI) (5 spot) um (r 2 > 0.5) using the PLINK software package. The subsequent and Ubiquitin. With the exception of Enolase, the expression (vol- analysis of the massive data generated by genome-wide SNP geno- ume %) of common spots differed between breeds, within the typing will pinpoint the genetic distinctiveness and levels of muscle. There were significative (P<0.05) differences of expres- admixture of Italian breeds, thus unraveling new aspects of the sion between breeds for 5 spots: Enolase, Myb and TPI (2 spots) for breed formation process. In addition, comparative analysis of con- the ‘Altamurana’; Ubiquitin for the ‘Gentile di Puglia’ and served haplotypes will identify genomic segments under selection ‘Leccese’. It was furthermore detected a different expression of pressure, highlighting interesting genomic regions. Data analysis some proteins between two breeding ‘bioterritories’: (i) higher will also be useful to select SNP loci suitable for parentage and expression of Myb and of some TPI spots in the breeds reared in identity DNA test in Italian breeds. the ‘Pugliese’ ‘bioterritory’; (ii) higher expression of DJ-1, Hb, PEBP and Ubiquitin proteins in the breeds reared in the ‘Sannita’ ‘bioterritory’. These differences might probably be ascribed to 9W environmental effects. Thus obtained results suggest a possible use of the proteomic approach to define physiological peculiarity Differential expression of sarcoplasmic proteins in in specific bioterritories; this may allow to establish a link meat of lambs reared in Continental Southern Italy between ovine meat and the territory that can be used to safe- guard local productions. Donato Matassino 1, Marzia Albenzio 2, Mariangela Caroprese 2, Elisabetta Castellana 3, Francesca Cecchi 4, Roberta Ciampolini 4, Dario Cianci 3, Elena Ciani 3, Mariasilvia D’Andrea 5, Carmen Di Prisco 1, Filomena Inglese 1, Rosaria Marino 2, Antonio Muscio 2, 10W Fabio Pilla 5, Francesco Romagnuolo 1, Antonella Santillo 2, 2 1,6 Ancient autochthonous genetic type Casertana pig: Agostino Sevi , Aldo Di Luccia influence of slaughter age on fatty acid composition 1Consorzio per la Sperimentazione, Divulgazione e Applicazione of subcutaneous adipose tissue di Biotecniche Innovative – Sub NFP.I.-FAO (Biodiversità 1 2 Mediterranea), Benevento, Italy Donato Matassino , Carmela Maria Assunta Barone , 1 1 1 2Dipartimento Scienze delle Produzioni e dell’Innovazione nei Carmen Di Prisco , Caterina Incoronato , Luigina Rillo , 1 2 3 Sistemi Agro-alimentari Mediterranei; 6Dipartimento di Scienze Giovanna Varricchio , Antonio Zullo , Aldo Di Luccia degli Alimenti, Università di Foggia, Italy 1Consorzio per la Sperimentazione, Divulgazione e Applicazione 3Dipartimento Fisiologia Generale ed Ambientale, Università di di Biotecniche Innovative – Sub NFP.I.-FAO (Biodiversità

[page 70] [Ital J Anim Sci vol.10:s1, 2011]