Analyzing the genetic variation of dairy sarda and spanish meat ovine breeds

The aims of this thesis were to investigate if variation at regulatory sites of the milk protein genes is associated with traits of economic interest in dairy Sarda sheep, as well as the genetic characterization of Spanish meat ovine breeds. Seven hundred and sixty Sarda sheep were genotyped with a multiplex TaqMan® assay of 29 SNPs found by us in the four casein genes and three previously reported SNPs in the LALBA and BLG genes. A high level of polymorphism (1 SNP every 200bp) was revealed and significant associations between CSN1S2 and CSN3 genotypes and milk protein and casein contents, while CSN1S1 and CSN2 genotypes were significantly associated with curd firming time. A total of 140 individuals belonging to the Xisqueta, Ripollesa, Segureña, Gallega, Roja Mallorquina and Canaria de Pelo breeds were genotyped with the Infinium 50K Ovine SNP BeadChip (Illumina). This dataset was merged with 50K genotypes of other five Spanish breeds (Churra, Ojalada, Castellana, Latxa and Rasa Aragonesa). Population genetics analyses revealed that Canaria de Pelo, Roja Mallorquina, Latxa and Churra were clearly differentiated populations. We performed a second experiment aiming to estimate the amount of shared vs breed-specific variation in five Spanish breeds through a RNA-seq approach. Our results showed a similar amount of shared vs pool-specific variants. Moreover, the number of pool-specific SNPs in Canaria de Pelo and Xisqueta was 2-5 fold higher than in the others breeds

The Effect of Long-term Freezing on Renneting Properties of Sarda Sheep Milk

Cold storage is a well-known strategy to limit concerns about seasonality for sheep dairy productions. The aim of this study was to investigate the effect of long-term freezing on milk renneting properties from the Sarda sheep, an autochthonous breed from Italy. Two-hundred milk samples from 50 pluriparous Sarda ewes were collected at monthly intervals throughout the lactation from April to July. Each sample of fresh milk was analysed for composition and subsamples were obtained and frozen for one, three and five months. Renneting properties, both from the fresh and frozen subsamples, were achieved using the Formagraph instrument and results were submitted to a mixed model statistical analysis. The storage effect significantly affected (P<0.01) the renneting parameters. A large amount of non coagulating subsamples was registered after a long-term frozen storage. Furthermore, milk clotting time was longer in frozen subsamples and curd firmness diminished after a freezing period of five months. In conclusion, the remarkable decreasing of sheep milk renneting characteristics after frozen storage can predict a worse yield and quality of cheese-making and suggests that freezing of Sarda raw milk should be limited to shorter periods

Titanium Functionalized with Polylysine Homopolymers: In Vitro Enhancement of Cells Growth

In oral implantology, the success and persistence of dental implants over time are guaranteed by the bone formation around the implant fixture and by the integrity of the peri-implant mucosa seal, which adheres to the abutment and becomes a barrier that hinders bacterial penetration and colonization close to the outer parts of the implant. Research is constantly engaged in looking for substances to coat the titanium surface that guarantees the formation and persistence of the peri-implant bone, as well as the integrity of the mucous perimeter surrounding the implant crown. The present study aimed to evaluate in vitro the effects of a titanium surface coated with polylysine homopolymers on the cell growth of dental pulp stem cells and keratinocytes to establish the potential clinical application. The results reported an increase in cell growth for both cellular types cultured with polylysine-coated titanium compared to cultures without titanium and those without coating. These preliminary data suggest the usefulness of polylysine coating not only for enhancing osteoinduction but also to speed the post-surgery mucosal healings, guarantee appropriate peri-implant epithelial seals, and protect the fixture against bacterial penetration, which is responsible for compromising the implant survival

The Effect of Long-term Freezing on Renneting Properties of Sarda Sheep Milk

Cold storage is a well-known strategy to limit concerns about seasonality for sheep dairy productions. The aim of this study was to investigate the effect of long-term freezing on milk renneting properties from the Sarda sheep, an autochthonous breed from Italy. Two-hundred milk samples from 50 pluriparous Sarda ewes were collected at monthly intervals throughout the lactation from April to July. Each sample of fresh milk was analysed for composition and subsamples were obtained and frozen for one, three and five months. Renneting properties, both from the fresh and frozen subsamples, were achieved using the Formagraph instrument and results were submitted to a mixed model statistical analysis. The storage effect significantly affected (P<0.01) the renneting parameters. A large amount of non coagulating subsamples was registered after a long-term frozen storage. Furthermore, milk clotting time was longer in frozen subsamples and curd firmness diminished after a freezing period of five months. In conclusion, the remarkable decreasing of sheep milk renneting characteristics after frozen storage can predict a worse yield and quality of cheese-making and suggests that freezing of Sarda raw milk should be limited to shorter periods

East African pigs have a complex Indian, Far Eastern and Western ancestry

In this study, we have characterized the mitochondrial diversity of 81 swine from Uganda. Median-joining network analysis of D-loop sequences from these individuals and others characterized in previous studies allowed us to determine that Ugandan pigs cluster with populations from the West (Europe/North Africa), Far East and India. In addition, partial sequencing of the Y-chromosome UTY locus in 18 Ugandan domestic pigs revealed the segregation of a single HY1 lineage that has a cosmopolitan distribution. A Western and Far Eastern ancestry for East African pigs had been already reported, but this is the first study demonstrating an additional contribution from the Indian porcine gene pool. This result is consistent with the high frequency of zebuine alleles in cattle from East Africa. The geographic coordinates of East Africa, at the crossroads of many trading routes that, through the ages, linked Europe, Africa and Asia, might explain the rich and complex genetic heritage of livestock native to this area.We are greatly indebted to the Swedish International Development Agency (Sida) under the framework of Sida-Mak Bilateral Research Support Programme Phase 3 to the Post Doc program at the Directorate of Graduate and Research Training for the financial support (Grant No. 75007369, Swedish Research Links, contract number 348-2011-7380).Peer reviewe

A genome-wide perspective about the diversity and demographic history of seven Spanish goat breeds

Altres ajuts: Félix Goyache is supported by Grant FICYT GRUPIN14-113. Valentin Balteanu is the recipient of a grant awarded under the frame of the European Social Fund, Human Resources Development Operational Program 2007-2013, Project No. POSDRU/159/1.5/S/132765.Background: The main goal of the current work was to infer the demographic history of seven Spanish goat breeds (Malagueña, Murciano-Granadina, Florida, Palmera, Mallorquina, Bermeya and Blanca de Rasquera) based on genomewide diversity data generated with the Illumina Goat SNP50 BeadChip (population size, N = 176). Five additional populations from Europe (Saanen and Carpathian) and Africa (Tunisian, Djallonké and Sahel) were also included in this analysis (N = 80) for comparative purposes. Results: Our results show that the genetic background of Spanish goats traces back mainly to European breeds although signs of North African admixture were detected in two Andalusian breeds (Malagueña and MurcianoGranadina). In general, observed and expected heterozygosities were quite similar across the seven Spanish goat breeds under analysis irrespective of their population size and conservation status. For the Mallorquina and Blanca de Rasquera breeds, which have suffered strong population declines during the past decades, we observed increased frequencies of large-sized (ROH), a finding that is consistent with recent inbreeding. In contrast, a substantial part of the genome of the Palmera goat breed comprised short ROH, which suggests a strong and ancient founder effect. Conclusions: Admixture with African goats, genetic drift and inbreeding have had different effects across the seven Spanish goat breeds analysed in the current work. This has generated distinct patterns of genome-wide diversity that provide new clues about the demographic history of these populations

Romanian wild boars and Mangalitza pigs have a European ancestry and harbour genetic signatures compatible with past population bottlenecks

We aimed to analyse the genetic diversity of Romanian wild boars and to compare it with that from other wild boar and pig populations from Europe and Asia. Partial sequencing of the mitochondrial encoded cytochrome b (MT-CYB) gene from 36 Romanian wild boars and 36 domestic pigs (Mangalitza, Bazna and Vietnamese breeds) showed that the diversity of Romanian wild boars and Mangalitza pigs is fairly reduced and that most of the members of these two populations share a common MT-CYB haplotype. Besides, in strong contrast with the Bazna animals, Romanian wild boars and Mangalitza swine did not carry Asian variants at the MT-CYB locus. The autosomal genotyping of 18 Romanian wild boars with the Illumina Porcine SNP60 BeadChip revealed that their genetic background is fundamentally European, even though signs of a potential Near Eastern ancestry (~25%) were detectable at K = 4 (the most significant number of clusters), but not at higher K-values. Admixture analysis also showed that two wild boars are of a hybrid origin, which could be explained by the mating of feral animals with domestic pigs. Finally, a number of Romanian wild boars displayed long runs of homozygosity, an observation that is consistent with the occurrence of past population bottlenecks and the raise of inbreeding possibly due to overhunting or to the outbreak of infectious diseases.This article was published under the frame of the European Social Fund, Human Resources Development Operational Program 2007–2013, project no. POSDRU/159/1.5/S/132765 awarded to VB. Part of the research presented in this publication was funded by grant AGL2013-48742-C2-1-R awarded by the Spanish Ministry of Economy and Competitivity to MA. We also acknowledge the support of the Spanish Ministry of Economy and Competitivity for the Center of Excellence Severo Ochoa 2016-2019 (SEV-2015-0533) grant awarded to the Center for Research in Agricultural Genomics.Peer reviewe

Romanian wild boars and Mangalitza pigs have a European ancestry and harbour genetic signatures compatible with past population bottlenecks

Altres ajuts: European Social Fund, Human Resources Development Operational Program 2007-2013, project no. POSDRU/159/1.5/S/132765We aimed to analyse the genetic diversity of Romanian wild boars and to compare it with that from other wild boar and pig populations from Europe and Asia. Partial sequencing of the mitochondrial encoded cytochrome b (MT-CYB) gene from 36 Romanian wild boars and 36 domestic pigs (Mangalitza, Bazna and Vietnamese breeds) showed that the diversity of Romanian wild boars and Mangalitza pigs is fairly reduced, and that most of the members of these two populations share a common MT-CYB haplotype. Besides, in strong contrast with the Bazna animals, Romanian wild boars and Mangalitza swine did not carry Asian variants at the MT-CYB locus. The autosomal genotyping of 18 Romanian wild boars with the Illumina Porcine SNP60 BeadChip revealed that their genetic background is fundamentally European, even though signs of a potential Near Eastern ancestry (∼25%) were detectable at K = 4 (the most significant number of clusters), but not at higher K-values. Admixture analysis also showed that two wild boars are of a hybrid origin, which could be explained by the mating of feral animals with domestic pigs. Finally, a number of Romanian wild boars displayed long runs of homozygosity, an observation that is consistent with the occurrence of past population bottlenecks and the raise of inbreeding possibly due to overhunting or to the outbreak of infectious diseases

Metabolic Pathway Modeling in Muscle of Male Marathon Mice (DUhTP) and Controls (DUC)-A Possible Role of Lactate Dehydrogenase in Metabolic Flexibility

In contracting muscles, carbohydrates and fatty acids serve as energy substrates; the predominant utilization depends on the workload. Here, we investigated the contribution of non-mitochondrial and mitochondrial metabolic pathways in response to repeated training in a polygenic, paternally selected marathon mouse model (DUhTP), characterized by exceptional running performance and an unselected control (DUC), with both lines descended from the same genetic background. Both lines underwent three weeks of high-speed treadmill training or were sedentary. Both lines' muscles and plasma were analyzed. Muscle RNA was sequenced, and KEGG pathway analysis was performed. Analyses of muscle revealed no significant selection-related differences in muscle structure. However, in response to physical exercise, glucose and fatty acid oxidation were stimulated, lactate dehydrogenase activity was reduced, and lactate formation was inhibited in the marathon mice compared with trained control mice. The lack of lactate formation in response to exercise appears to be associated with increased lipid mobilization from peripheral adipose tissue in DUhTP mice, suggesting a specific benefit of lactate avoidance. Thus, results from the analysis of muscle metabolism in born marathon mice, shaped by 35 years (140 generations) of phenotype selection for superior running performance, suggest increased metabolic flexibility in male marathon mice toward lipid catabolism regulated by lactate dehydrogenase.Peer reviewe

Expression patterns and genetic variation of the ovine skeletal muscle transcriptome of sheep from five Spanish meat breeds

The goal of the current study is to analyse the gene expression profile of the ovine skeletal muscle as well as to characterize the genetic variation of transcripts expressed in such tissue. This aim has been achieved by sequencing the longissimus dorsi transcriptomes of 50 sheep distributed in five pools representing the Canaria de Pelo, Roja Mallorquina, Gallega, Xisqueta and Ripollesa Spanish autochthonous breeds. Approximately, 363 million reads per pool have been produced and 71.9-82.9% have been successfully mapped to the ovine genome in a paired-end mode (2 X 75 bp). The 200 most expressed muscle transcripts (-1% of the total transcript count) account for 51% (Canaria de Pelo) to 67% (Gallega) of the total ovine skeletal muscle mRNA expression. These highly expressed genes play key roles in pathways related with striated muscle contraction, gluconeogenesis, glycolysis, citric acid cycle and respiratory electron transport. RNA-Sequencing of muscle transcripts has also revealed that -72% of the SNPs detected with this approach are shared by at least two pools, and 10% of them segregate in the five pools under analysis. Most of the substitutions detected by RNA-Seq are synonymous or missense and only a minority are predicted to have consequences on protein function