[Show abstract][Hide abstract] ABSTRACT: The Hispano-Breton equine breed is located in the North of the Iberian Peninsula and currently is an endangered breed. In order to analyze the genetic variability of the Hispano-Breton horse breed and its genetic relationships with other horse breeds located in the Iberian Peninsula a total of 25 autosomal microsatellites have been analyzed in 30 samples of the Hispano-Breton horse breed and in an additional 20 horse breeds that represent a comprehensive sampling of current Iberian Peninsula horse breeds. The parameters of genetic diversity of the Hispano-Breton horse breed were in the upper range of the values estimated in all of the breeds analyzed. The average FST genetic distance of Hispano-Breton horse with respect to the other breeds was 0.06. It is remarkable that the average FST genetic distance among Hispano-Breton, Burguete and Pirinenc Català horse breeds was not significantly different from 0. The FIS estimate in the Hispano-Breton did not indicate a significant deficit of heterozygotes. The dendrogram built from FST genetic distances among breeds and the correspondence analysis showed similar results. The Iberian Peninsula celtic horse breeds were grouped in a single cluster while the horse breeds with different degrees of Arab influence comprise a second cluster. The low census and corresponding genetic drift effect probably explain the isolation evidenced by the Sorraia and Retuertas horse breeds.
[Show abstract][Hide abstract] ABSTRACT: The knowledge of the genetic composition and relationships among livestock breeds is a necessary step for the implementation of management and conservation plans. This study aims to characterise the genetic diversity and relationships among six sheep breeds of meat aptitude that are spread through the eastern Pyrenees: Tarasconnaise, Castillonnaise and Rouge du Roussillon from France, and Aranesa, Xisqueta and Ripollesa from Spain. All but Tarasconnaise are catalogued as endangered. These breeds do not share the same ancestral origin but commercial trades and gene flow between herds are known to have occurred for centuries. Additionally, two outgroup breeds were included: the Guirra, from a different geographical location, and the Lacaune, a highly selected breed of dairy aptitude. A total of 410 individuals were typed using a panel of 12 microsatellite markers. Statistical, phylogenetic and Bayesian analyses showed that eastern Pyrenean breeds retained high levels of genetic diversity and low, but significant, levels of genetic differentiation (FST = 4.1%). While outgroups were clearly differentiated from other breeds, Pyrenean breeds tended to form two clusters. The first encompassed Tarasconnaise and Aranesa, which probably descend from a common meta-population. The second tended to group the other four breeds. However, none reached high mean Q-values of membership to a discrete cluster. This is consistent with the recent past gene flow between breeds, despite different ancestral genetic origins. The genetic characterisation carried out of the eastern Pyrenean sheep populations provides useful information to support decision making on their conservation and focusing efforts and resources to more singular breeds.
SPANISH JOURNAL OF AGRICULTURAL RESEARCH 12/2014; 12(4):1029-1037. DOI:10.5424/sjar/2014124-6173 · 0.51 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The adiponectin (ADIPOQ) locus is a positional and functional candidate gene for two porcine chromosome 13 (SSC13) QTL influencing cholesterol (CHOL) and low density lipoprotein (LDL) concentrations in 190 days-old pigs. By sequencing 2.37 kb of the pig ADIPOQ cDNA, we have identified one c.*1512G>T 3'UTR polymorphism that has been genotyped in a Duroc pig commercial population with records for serum lipid levels at 45 and 190 days-of-age. Statistical analysis of the data have revealed significant associations between ADIPOQ genotype and CHOL (P = 0.0040) and LDL (P = 0.0011) concentrations at 190 days, but not at 45 days. In family 3, most part of the SSC13 QTL effects on LDL levels at 190 days were explained by the ADIPOQ genotype. We also found an association with triglyceride levels at 45 days (P = 0.0060), but not at 190 days. Measurement of allelic mRNA imbalance demonstrated that the G- and T-alleles are expressed at very similar levels at muscle and fat tissues, indicating that the c.*1512G>T polymorphism does not affect transcript abundance. As a whole, results obtained in the current work, as well as previous data gathered in humans and pigs, evidence that the magnitude of associations between blood lipid phenotypes and candidate loci genotypes may vary depending on the age of the individual, thus suggesting the existence of dynamic genotype by environment interactions changing on a temporal scale.
[Show abstract][Hide abstract] ABSTRACT: Inferring the breed of origin of dairy products can be achieved through molecular analysis of genetic markers with a population-specific pattern of segregation. The goal of the current work was to generate such markers in goats by resequencing several pigmentation genes [melanocortin 1 receptor (MC1R), v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (KIT), tyrosinase (TYR), and tyrosinase-related protein 2 (TYRP2)]. This experiment revealed 10 single nucleotide polymorphisms (SNP), including 5 missense mutations and 1 nonsense mutation. These markers were genotyped in 560 goats from 18 breeds originally from Italy, the Iberian Peninsula, the Canary Islands, and North Africa. Although the majority of SNP segregated at moderate frequencies in all populations (including 2 additional markers that were used as a source of information), we identified a c.764G>A SNP in MC1R that displayed highly divergent allelic frequencies in the Palmera breed compared with the Majorera and Tinerfeña breeds from the Canary Islands. Thus, we optimized a pyrosequencing-based technique that allowed us to estimate, very accurately, the allele frequencies of this marker in complex DNA mixtures from different individuals. Once validated, we applied this method to generating breed-specific DNA profiles that made it possible to detect fraudulent cheeses in which Palmero cheese was manufactured with milk from Majorera goats. One limitation of this approach, however, is that it cannot be used to detect unlawful manufacturer where Palmero dairy products are produced by mixing milk from Palmera and Majorera goats, because the c.764G>A SNP segregates in both breeds.
[Show abstract][Hide abstract] ABSTRACT: When artificial reproduction technologies designed for use with horses are used with donkeys, success is dependent on awareness of the physiological differences between these species, yet little information is available on many aspects of donkey reproduction. The present work examines the activity of the corpus luteum (CL) in Catalonian jennies following induced luteolysis. Plasma progesterone concentration, luteal blood flow (determined by colour-Doppler) and CL cross-sectional area (CL-CSA; determined by B-mode ultrasound examination) were assessed following a single dose ( 5mg i.m.) of dinoprost thromethamine (DT - a PGF2α analogue) on day 10 post-ovulation in two experiments. In Experiment 1 – a preliminary experiment - data were collected daily for 4 days following DT administration. Values for all the measured variables decreased over this period. In Experiment 2, data were collected during the first 24 h following DT administration since, in Experiment 1, most luteolytic activity occurred during this time. An increase in luteal blood flow was seen between 0 and 3 h, followed by a progressive reduction, while the values for plasma progesterone and CL-CSA gradually decreased from 0 h onwards. In both studies, negative correlations were seen between all variables and the time of sampling. In contrast, positive correlations were seen between plasma progesterone, CL-CSA, uterine tone and luteal blood flow. Indeed, a strong correlation was recorded between plasma progesterone and luteal blood flow (r=0.70; P<0.0001). In conclusion, plasma progesterone and CL-CSA both become reduced after induced luteolysis in Catalonian jennies. Unlike in mares, an increase in luteal blood flow occurs soon after induced luteolysis, rather like that seen in the cow. The luteal blood flow, as evaluated here by colour-Doppler, was also closely related to the plasma progesterone concentration. Colour-Doppler would appear therefore to offer a rapid and easy means of examining the state of luteolysis.
[Show abstract][Hide abstract] ABSTRACT: The CD36 molecule plays a key role in the uptake of long-chain fatty acids. In a previous study, we demonstrated that the CD36 gene is duplicated in goats. Moreover, both copies (CD36 and CD36-like) display highly divergent mRNA expression profiles. Herewith, we have analysed wether four polymorphisms mapping to CD36 (c.394A > G, c.*141C > T and c.*427T > A) and CD36-like (c.390A > C) genes are associated with milk yield and composition. Murciano-Granadina goats with records for dairy traits (N = 309, 1005 registers) and milk fatty acid composition phenotypes (N = 176, 490 registers) were genotyped for these four markers and association analyses were carried out. We found a highly significant association between c.*427T > A CD36 polymorphism and milk palmitoleic content (P < 0.0001). Besides, the c.*141C > T CD36 SNP also showed a suggestive association (P = 0.04) with palmitoleic. These findings are consistent with previous studies showing that CD36 inactivation in adipocytes and neurons involves a decrease in palmitoleic content, thus suggesting a relevant role of this molecule in the specific uptake of this fatty acid. The remaining associations found (CD36 with polyunsaturated fatty acids and CD36-like with oleic and several saturated fatty acids) were significant at the nominal level but not after Bonferroni correction. These results, combined with previously reported expression data, reinforce the interest of investigating the lipid binding properties of CD36 and CD36-like through functional approaches.
Small Ruminant Research 07/2014; 121(2-3). DOI:10.1016/j.smallrumres.2014.07.008 · 1.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To evaluate the effect of seminal plasma in endometrial inflammation in donkeys, samples from fresh pure, fresh diluted and frozen-thawed semen of three different jackasses were co-incubated in water bath at 37°C with uterine Jennie's secretions collected 6h after artificial insemination with frozen-thawed donkey semen. Individual sperm movement parameters using the computerised sperm analysis system (CASA) and sperm-polymorphonuclear neutrophils (sperm-PMN) attachment observed in Diff-Quick stained smears were evaluated at 0, 1, 2, 3 and 4h of co-incubation. Controls consisted of incubating diluted or frozen-thawed sperm in the absence of uterine secretions. For data analyses, a repeated measures ANOVA was performed with incubation time as intra-subject factor and with treatment and donkey as inter-subject factor, followed by a post-hoc Bonferroni's test. Greater values (P<0.05) of sperm-PMN percentages and a loss of progressive motility were observed in frozen-thawed semen compared with pure and diluted fresh semen samples throughout the incubation time. In addition, the presence of seminal plasma in fresh and diluted semen samples reduced the inflammatory response of polymorphonuclear neutrophils produced after insemination by suppressing the sperm-PMN attachment in vitro. Motility sperm parameters analysed by CASA were also less affected than those in frozen-thawed semen samples. In conclusion, seminal plasma in jennies appears to have a modulation on the endometrial response after artificial insemination with frozen-thawed donkey semen. As a result, spermatozoa with the greater motility characteristics are selected.
[Show abstract][Hide abstract] ABSTRACT: The performance of a genome scan for serum lipid traits at 45 and 190 days in five half-sib families of Duroc pigs allowed us to detect several pig chromosomal regions with significant effects on serum lipid traits at 45 and 190 days. In the current work, we aimed to refine the position of one chromosome 4 (SSC4) genome-wide significant QTL for serum triglyceride concentration at 190 days. Genotyping of four additional microsatellites allowed to reduce the 90% CI of this QTL to the genomic interval comprised between markers SW2409 and SW839. Sequencing experiments were carried out to characterize the variability of two lipid-related genes, the lipoprotein receptor-related protein 12 (LRP12) and tribbles homolog 1 (TRIB1) loci, that map to this region. In this way, two (c.771A>G and c.1101A>G) and one (c.*156_157del) polymorphisms were identified at the LRP12 coding region and TRIB1 3'UTR, respectively. Association analyses between LRP12 and TRIB1 genotypes did not reveal any significant effect on serum lipid levels, suggesting that their variation does not explain the segregation of the SSC4 QTL. However, highly significant associations were observed for gluteus medius saturated fatty acid content (LRP12 c.1101A>G, P = 0.0006; TRIB1, c.*156_157del, P = 0.0003). In the light of these and other findings, the potential involvement of LRP12 and TRIB1 in muscle lipid metabolism deserves to be further explored.
[Show abstract][Hide abstract] ABSTRACT: In an attempt to increase the possibilities of a better sperm cryopreservation from these endangered small ruminant catalonian breeds, we are studying different strategies. One of them is to study the effect of the buffer system Tes-Tris (TEST) compared with the Tris and citric acid (TRIS) buffer system, testing simultaneously both systems in a 1% (w/v) soybean lecithin or in a 15% (v/v) powdered egg yolk-based media supplemented both with 5% glycerol and to assess also the inclusion of 5mM of butylated hydroxytoluene (BHT) as an antioxidant on the cryopreservation media. However, the main objective of the present study was to test first the effect of the different media when the sperm were cultured 4h at 5°C. Briefly, fresh ejaculates from 6 young bucks of Blanca de Rasquera breed (1 year old) were collected by an artificial vagina in favourable reproductive period and immediately mixed in equal quantities. Pooled ejaculates were split into 2 equal aliquots and washed by centrifugation (twice for 10min at 600g) in TRIS- or TEST-based media without cryoprotectants and antioxidant. Afterward each pellet was split into 4 equal aliquots, re-suspended in TRIS or TEST, depending on the experimental group, soybean lecithin, or powdered egg yolk-based media, and supplemented or not with BHT and kept for 4h at 5°C. Likewise, fresh ejaculates from 8 young rams (4 rams of Xisqueta and 4 rams of Aranesa breed, 1 year old) were collected and processed as buck semen samples. Sperm survival before cryopreservation was determined by eosine-nigrosine stain, and sperm motion parameters were analysed by a computer-assisted sperm analysis system (ISAS(®)). Six replications were performed in both species, and General Lineal Model (SAS(®), Cary, NC, USA) was used for the statistical analysis. The highest sperm viability percentage (mean ± SE) on goat sperm cultured 4h at 5°C was observed in the extender with TRIS buffer system in powdered egg yolk-based media supplemented with BHT (81.1±2.8), not showing significant differences with the others extenders, except with the viability of the samples in the extenders with TEST buffer system in soybean lecithin-based media supplemented (56.5±2.9; P<0.001) or not with BHT (60.1±5.1; P<0.001). On the other hand, no significant differences on sperm viability were observed on ram sperm between treatments. Nevertheless, the sperm quality motion characteristics (data not shown) were quite different between all the treatments in both species. Considering that the present results are still preliminary, we suggest that more analysis should be made to explain how the different composition of the extenders affect sperm quality during the cryopreservation process.
Reproduction Fertility and Development 12/2012; 25(1):184. DOI:10.1071/RDv25n1Ab73 · 2.58 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The CD36 molecule plays a pivotal role in a variety of immunological and cellular processes, including pathogen recognition, inflammation and apoptosis. Herein, we demonstrate that this gene is duplicated in goats, with two copies (CD36 and CD36-like) that display highly divergent mRNA expression profiles. In this way, CD36 mRNA is mostly expressed in the adipose tissue and heart whilst CD36-like mRNA shows a high expression in the liver. We have also found evidence of the presence of two paralogous CD36 and CD36-like genes in the bovine genome, suggesting that CD36 duplication took place before goat-cattle radiation, i.e. at least 20 MYR ago. Finally, we have characterized the polymorphism of the coding regions of the goat CD36 and CD36-like genes. In doing so, we have identified one synonymous polymorphism at the CD36-like gene (c.390A>C) that displays a significant association (P=0.04) with milk somatic cell count, a parameter often used to diagnose mastitis in domestic ruminants.
[Show abstract][Hide abstract] ABSTRACT: Manunza, A., Zidi, A., Fernandez-Cabanas, V. M., Jordana, J., Carrizosa, J., Belaifa, E., Urrutia, B., Polvillo, O., Gonzalez-Redondo, P., Amills, M. and Serradilla, J. M. 2012. SHORT COMMUNICATION: An association analysis between one missense polymorphism at the SREBF1 gene and milk yield and composition traits in goats. Can. J. Anim. Sci. 92: 167-173. Sterol regulatory element binding transcription factor 1 (SREBF1) regulates the expression of genes involved in the biosynthesis of fatty acids and cholesterol. Herewith, we have sequenced the near-complete coding region and part of the 3'UTR of the goat SREBF1 gene. In doing so, we have detected a missense c.353C>T polymorphism causing a proline to leucine substitution at position 118 (P118L). An association analysis with milk composition traits recorded in Murciano-Granadina goats only revealed a statistical tendency linking SREBF1 genotype and milk omega-3 fatty acid content. The lack of significant associations suggests that the P118L substitution does not involve a functional change.
Canadian Journal of Animal Science 06/2012; 92(2):167-173. DOI:10.4141/CJAS2011-111 · 0.98 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Demographic processes leave specific genetic signatures that can be detected through the analysis of nuclear and mitochondrial markers. Often, these signatures are very complex to interpret due to the co-occurrence of processes that are very distinct in nature and magnitude. In this work, we have analysed the footprint of past bottlenecks in the gene pool of Azpi Gorri goats, a Basque breed in a critical status of conservation because of dramatic reductions in its census. A sample of Azpi Gorri goats was analysed by sequencing the mitochondrial control region (N = 24) and genotyping 29 autosomal microsatellites (N = 50). Contrary to our expectations, the analysis of nuclear and mitochondrial markers showed that the Azpi Gorri breed has retained a considerable level of genetic diversity. Even more, conflicting results were obtained when trying to detect bottleneck genetic signatures through the calculation of R2 and Fu's FS statistics (mitochondrial data) and by performing Bottleneck and M_P_VAL analyses (microsatellite data). Cryptic bottleneck genetic signatures might be relatively frequent in domestic species because population declines are usually smooth and sustained, rather than drastic and instantaneous, and also because many concomitant factors, such as population subdivision, selection and migration, can alter their shape.
Small Ruminant Research 05/2012; 104(s 1–3):78–84. DOI:10.1016/j.smallrumres.2011.11.012 · 1.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Spanish heavy horses play a very important role in keeping up ecologically important rural areas. The interest in their conservation and characterization has increased over the last few years. The aim of this work is to contribute to the characterization of the endangered Spanish heavy horse populations in order to obtain useful information to implement conservation strategies for these genetic stocks. A total of 426 horses from six Spanish heavy horse populations corresponding to the four main heavy horse breeds (Hispano-Breton (HB) – with most animals located in three areas: Burgos (HBbu), León (HBle) and Palencia (HBpa) – Jaca Navarra (JN), Burguete (B) and the Cavall Pirinenc Català (CPC)) were analysed using 22 body measurements, 10 indices calculated from the body measurements and 16 microsatellite markers. From a morphological point of view, all the populations are clearly differentiated, except for the HBpa female population, which is located between the HBbu and JN populations. The HBle has greater body size than the other populations analysed, especially when compared to the JN. The genetic analysis provides us with information about the history of the analysed breeds. However, the most important point borne out by this analysis is the clear identification of different genetic backgrounds within the Spanish heavy horse populations. The overall information given in this work leads us to consider that the genetic scenario of the Spanish heavy horses is more likely to be due to ‘ancestrally’ different genetic backgrounds. Spanish heavy horse breeds resulted from the crosses between native mares and foreign stallions. The CPC population was the first in which this introgression occurred and the use of foreign stallions became more common. This work presents results based on neutral genetic variation, but also within- and between-population differences in morphological traits that have undergone artificial or natural (adaptive) selection. This information should be taken into account in future conservation strategies in order to contribute towards the efficiency of conservation measures. In this sense, the main problems are the small population size and genetic degeneration, with the consequent future loss of diversity. Therefore, the most immediate and effective conservation priorities would be (1) to avoid inbreeding within populations, (2) to increase the population size, and (3) to facilitate genetic exchange among the populations.
[Show abstract][Hide abstract] ABSTRACT: Tyrosinase-related protein 1 (TYRP1) has been shown to play a fundamental role in pigmentation both in human and mouse. In this work, we aimed to characterize the variability of the caprine TYRP1 gene and investigate its segregation in a wide array of goat breeds. By partially sequencing the coding region of the TYRP1 gene in 18 individuals from eight different breeds, we were able to identify a synonymous nucleotide substitution at exon 3 (c.483C>T). An extensive survey of Iberian and Balearic (N=175), Italian (N=99), Swiss (N=54), Asian (N=14), Canarian (N=92) and North African (N=117) goats with different coat colours was carried out. We found that the C-allele has a different distribution in European vs African breeds, being almost fixed in the latter. Moreover, the C-allele showed an increased frequency in white coated breeds (Girgentana, Grigia Molisana, Blanca de Rasquera and Saanen) when compared with those displaying a dark pigmentation (Cilentana Nera, Azpi Gorri and Murciano-Granadina). This could be due to genetic drift, migration and other factors associated with the demographic history of breeds under analysis or to a genetic hitchhiking event (c.483C>T frequencies would be shaped by a neighbouring causal mutation differentially selected in white and black goats). More refined studies will be needed to distinguish between these two alternative explanations.
[Show abstract][Hide abstract] ABSTRACT: En el marco de un Programa de Conservación, se presentan los resultados de la caracterización productiva de la raza caprina Blanca de Rasquera, de aptitud cárnica. Se estudiaron un total de 125 animales escogidos de forma aleatoria y distribuidos en 6 explotaciones de régimen extensivo y lactancia natural. Se caracterizó el crecimiento de los cabritos mediante la cuantificación de los pesos desde el nacimiento hasta el sacrificio. Se evaluó la influencia del sexo y tipo de parto sobre el crecimiento. Los cabritos presentaron una buena tasa de crecimiento (123 g/día como promedio, para el período 0-45 días de edad), comparable, o ligeramente inferior, a otras razas cárnicas autóctonas españolas. Como era esperable, el tipo de nacimiento y el sexo influyeron de forma significativa en el peso y el crecimiento de los cabritos. Se analizó una muestra aleatoria de 36 individuos, en relación al gen de la αS1-caseína (CSN1S1). Las mayores frecuencias alélicas (62,5%) se encontraron para los alelos asociados con una elevada cantidad de αS1-caseína en la leche, indicando dichos resultados las buenas aptitudes y cualidades tecnológicas de su leche para la producción de quesos.