D. Cianci

The genetic variability of the Podolica cattle breed from the Gargano area. Preliminary results

Abstract The Podolica cattle breed is autochthonous of Southern Italy and denoted by its particular rusticity. This study presents the preliminary results of the genetic characterization of the Podolica breed using DNA STR markers. A total of 20 microsatellite loci were analysed in 79 individuals reared in the Gargano area. Number of polymorphisms, allele frequencies, deviations from Hardy-Weinberg proportions, linkage disequilibrium between loci and genetic similarities between animals were calculated. The results showed a high deficiency of heterozygotes, the observed mean of heterozygosis being 0.449, whereas the expected mean was 0.766. Many markers showed also deviations from the Hardy-Weinberg proportions and significant linkage disequilibrium between loci. However the genetic similarity within the population was low (0.281) and the average number of alleles per locus was high (10), representing a high genetic variability. In order to explain these results, a stratification of the breed in sub-populations with a high interior genetic homogeneity but markedly differentiated one from each other could be hypothesized; this situation probably derived from non-random mating within each herd (consanguinity) and from the lack of exchange of genetic material between the herds. A further study is needed on a wider sample and extending the analysis to FAO-ISAG microsatellite panel in order to confirm this hypothesis. This could eventually provide the information necessary for the correct management of the reproductive schemes and for genomic traceability of meat production.

Haplotype association analysis of meat quality traits at the bovine PRKAG3 locus

Abstract The current study presents the results of a preliminary haplotype association analysis at the bovine PRKAG3 locus with meat quality traits in the Chianina breed. No significant association was shown between haploid haplotypes (or diplotypes) and phenotypical traits after applying a Bonferroni correction for multiple comparison. Nonetheless, data from Longissimus dorsi muscle suggest the presence of a statistically non-significant trend toward an influence of the PRKAG3 haploid haplotypes on meat colour (a*) and water holding capacity (M/T) traits, as confirmed also by diplotype-based association analysis. A less clear set of results was observed for the Triceps brachii and Semitendinosus muscles.

Genetic variability of three local cattle breeds (Calvana, Pontremolese, Garfagnina) by STR analysis

Abstract The dramatic size contraction of local cattle breeds due to replacement with cosmopolite improved breeds highlights the need for native genetic resources conservation. In 1985, the Anagraphic Register of local cattle breeds and small-size ethnic groups was established by the Italian Ministry of Agriculture and Forestry. Calvana, Pontremolese and Garfagnina are among the included breeds. They are all native from Tuscany. Present breeding area covers the provinces of Firenze, Prato, Pistoia and Siena for the Calvana breed (around 280 heads), while it is restricted to the province of Lucca for both Garfagnina (around 180 heads) and Pontremolese. This latter breed consists, nowadays, of less than 40 heads, while being around 15000 in 1940s. The characterization of the genetic structure and variability via molecular markers could provide useful information for breed management and conservation. In the present study, a total of 149 animals, evenly distributed among the three breeds, were genetically characterized by using 22 STR markers located on 13 different chromosomes. Among these, ten (BM1818, BM1824, ETH10, ETH152, ETH3, HEL9, ILST006, INRA63, TGLA126, TGLA227) belonged to the panel of 30 microsatellites recommended by the ISAG-FAO Working Group on domestic animal diversity. Mean number of alleles per locus in the total sample was 9.9, ranging from 5 (ETH10) to 14 (BMS690, HEL9, ETH152). Within population mean number of alleles did not vary among the three population samples (range, 6.4 to 6.8). On the other hand, almost all loci showed several low frequency alleles “private” to one breed only. Mean locus heterozygosity in the total sample was 0.426. All loci showed a significant excess of homozygous genotypes in all the three breeds. Only the HEL9 locus (BTA8) was characterized, in all breeds, by a significant excess of heterozygous genotypes. Average FIS over all loci was 0.287 for Garfagnina, 0.378 for Calvana, and 0,479 for Pontremolese. Significant pair-wise linkage disequilibrium (or gametic imbalance, for unlinked loci) was observed for all breeds. In particular, more than 40% of all possible pair-wise comparisons were in linkage disequilibrium in the Pontremolese breed. A breed assignment test based on a log-likelihood approach allowed the correct allocation of 100% of individuals to their true breed of origin.

The genetic resistance to gastro-intestinal strongylids in Appenninica sheep: relationship among parasitical load and haematological parameters

Abstract Faecal Egg Count (FEC) is generally considered a simple but effective parameter for estimating resistance to strongylids, due to its phenotypic relationship to the total number of parasites present in the animal. Nevertheless, also haematological indicators have been shown to be useful in measuring gastro-intestinal nematode resistance in sheep, such as Packed Cell Volume (PCV), number of leukocytes, proportion of eosinophils and plasma albumin. The study was conducted in a single herd of Appenninica sheep situated in Tuscany (Italy) with the aim of studying the relationship among parasitic load of gastro-intestinal Strongylids and haematological parameters. Faecal samples from 54 sheep were collected in four different withdrawals (April, May, September and November 2004) and analysed in order to determine parasite EPG (Eggs Per Gram), using the McMaster technique. At the same time as the faecal samples, blood samples were also collected in order to determine PCV, total proteins, complement proteins, plasma albumin, erythrocytes and eosinophils. Considering the low infection level in spring (April and May), it was believed that animals with a positive EPG in this season could already be considered “not resistant”, although a limited infection (EPG<200) might serve as an index of host’s capacity to keep the parasite population under control. Consequently, we defined as “resistant group” (RG) the subjects with an average value in the spring sample equal to zero EPG, “intermediate resistant group” (IRG) the subjects with an average value under 200 EPG and “not resistant group” (NRG) the animals with an average value over 200 EPG. NRG subjects always showed lower values of PCV (an indicator of anaemia) than the other groups. In September, NRG subjects also showed significantly lower values of eosinophils (3.28%) compared to the other groups (6.54 and 6.20%, respectively, for IRG and RG, P<0.01) and higher values of complement (20.06 vs. 12.61 and 12.77, respectively, for IRG and RG, P<0.05). On the contrary, in November, NRG subjects showed lower values of complement (7.78) than the other groups (19.00 and 15.23, respectively, for IRG and IR, P<0.01).

Associations Between Microsatellite Markers and Milk Traits in Massese Sheep: Preliminary Results

Carnitine is a small essential molecule that regulates the substrate flux and energy balance across cell membranes by modulating both the transport of long-chain fatty acids into mitochondria and their subsequent β-oxidation. Although humans are capable to synthesize it endogenously, approximately 75% of body carnitine sources come from diet and particularly from food of animal origin such as meat, poultry, fish and dairy products. Due to its intrinsic interaction with the bioenergetics processes, carnitine plays an important role in diseases associated with metabolic compromise, especially mitochondria-related disorders. It has been reported that administration of carnitine by diet or at pharmacological doses can have significant benefit in several physiopathological situations such as ischemia, myocardial injury and neurodegenerative diseases, but there is no data on the possible protective role of carnitine against other oxidative stress-induced pathologies associated with an altered chromosome stability such as cancer. Therefore, we analysed the potential capability of carnitine to protect mammalian cells from genetic instability induced by H2O2, using Chinese Hamster Ovary (CHO) cells as a mammalian cell model having a stable karyotype and the chromosome aberration test as genetic end point. Our results showed that in the absence of carnitine H2O2 induced a high and dose-dependent induction of structural chromosome aberrations in the concentration range 0.1-0.4 mM whereas at the same H2O2 doses, a pre-treatment with 4 mM carnitine produced a strong decrease either of the percent of cells with aberrations or of the aberration frequency. The observed carnitine-mediated prevention of H2O2-induced chromosome aberrations reaches almost the control value in the cultures treated with 0.1 mM of H2O2 thus evidencing a reduction of about 70%. These data, together with preliminary results showing that carnitine is not able to protect cells from the inhibition of cell growth caused by H2O2, suggest that carnitine protects mammalian cells from H2O2–induced clastogenic damage and this effect is reproducible and highly specific.

Analysis of genetic differentiation of a sheep population by Cluster Analysis on milk serum proteins: preliminary results

Carnitine is a small essential molecule that regulates the substrate flux and energy balance across cell membranes by modulating both the transport of long-chain fatty acids into mitochondria and their subsequent β-oxidation. Although humans are capable to synthesize it endogenously, approximately 75% of body carnitine sources come from diet and particularly from food of animal origin such as meat, poultry, fish and dairy products. Due to its intrinsic interaction with the bioenergetics processes, carnitine plays an important role in diseases associated with metabolic compromise, especially mitochondria-related disorders. It has been reported that administration of carnitine by diet or at pharmacological doses can have significant benefit in several physiopathological situations such as ischemia, myocardial injury and neurodegenerative diseases, but there is no data on the possible protective role of carnitine against other oxidative stress-induced pathologies associated with an altered chromosome stability such as cancer. Therefore, we analysed the potential capability of carnitine to protect mammalian cells from genetic instability induced by H2O2, using Chinese Hamster Ovary (CHO) cells as a mammalian cell model having a stable karyotype and the chromosome aberration test as genetic end point. Our results showed that in the absence of carnitine H2O2 induced a high and dose-dependent induction of structural chromosome aberrations in the concentration range 0.1-0.4 mM whereas at the same H2O2 doses, a pre-treatment with 4 mM carnitine produced a strong decrease either of the percent of cells with aberrations or of the aberration frequency. The observed carnitine-mediated prevention of H2O2-induced chromosome aberrations reaches almost the control value in the cultures treated with 0.1 mM of H2O2 thus evidencing a reduction of about 70%. These data, together with preliminary results showing that carnitine is not able to protect cells from the inhibition of cell growth caused by H2O2, suggest that carnitine protects mammalian cells from H2O2–induced clastogenic damage and this effect is reproducible and highly specific.