Open Access
Expression of key myogenic, fibrogenic and adipogenic genes in Longissimus thoracis and masseter muscles in cattle
(Cambridge University Press, 2020) Martínez del Pino, Lara; Urrutia Vera, Olaia; Arana Navarro, Ana; Alfonso Ruiz, Leopoldo; Mendizábal Aizpuru, José Antonio; Soret Lafraya, Beatriz; Agronomia, Bioteknologia eta Elikadura; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD; Agronomía, Biotecnología y Alimentación
Adipogenesis, myogenesis and fibrogenesis are related processes that can contribute to meat quality. Therefore, extending the knowledge of these processes would facilitate the identification of molecular markers that predict intramuscular fat accretion. The main purpose of this work, based on previous results, was to further study the expression of key genes related to adipogenic, myogenic, fibrogenic processes and some cytokines in Longissimus thoracis (LT) and Masseter (MS) muscles of Pirenaica and Holstein young bulls. Longissimus thoracis and MS muscles from Pirenaica (n = 4) and Spanish Holstein (n = 4) were sampled for proximate analysis, determination of adipocyte size distribution and expression of key candidate genes. Fat percentage was lower in LT than in MS muscle in Pirenaica young bulls (P = 0.023) and was higher in LT muscle in Holstein than in Pirenaica young bulls (P = 0.007). Gene expression analysis revealed that the mRNA level of myogenic differentiation 1 (MYOD) was higher in LT than in MS muscles in both groups of animals (P < 0.001) and that myostatin (MSTN) expression was also higher in LT than in MS muscle in Holstein bulls (P = 0.001). On the other hand, MSTN and PPARG showed higher expression in LT and MS in Pirenaica young bulls (P = 0.026), while the expression of fatty acid-binding protein 4 (FABP4) was higher in Holstein young bulls, also in both muscles (P < 0.001). The results suggested that the development of intramuscular adipose depot was directly related to the expression of adipogenic genes, such as FABP4, but inversely related to the expression of the cytokine MSTN and the myogenic gene MYOD, genes which showed a muscle-specific expression.
Open Access
Mipob: un programa de simulación para el aprendizaje en mejora genética animal
(Universidad de Córdoba, 2014) Alfonso Ruiz, Leopoldo; Producción Agraria; Nekazaritza Ekoizpena
Se ha desarrollado un programa de simulación de apoyo a la docencia de los cursos introductorios a la Mejora genética animal. El programa surgió ante la ausencia de este tipo de herramientas docentes orientadas a estudiantes de primeros cursos de grado universitario, especialmente en lengua española. El programa simula una población animal cerrada de censo reducido en la que los estudiantes deben de ir tomando las decisiones de elección de futuros reproductores y cómo aparearlos. De esta forma se facilita la introducción de los conceptos básicos sobre selección y evaluación genética, y conservación y genética de poblaciones. El programa considera el caso particular del vacuno de carne y se pueden realizar hasta un total de diez generaciones de selección. Tras haber sido probado durante dos cursos consecutivos en enseñanza reglada universitaria, está disponible de forma gratuita para su libre utilización y distribución.
Open Access
Adiposity and adipogenic gene expression in four different muscles in beef cattle
(Public Library of Science, 2017) Martínez del Pino, Lara; Arana Navarro, Ana; Alfonso Ruiz, Leopoldo; Mendizábal Aizpuru, José Antonio; Soret Lafraya, Beatriz; Producción Agraria; Nekazaritza Ekoizpena
Anatomical site and divergent functionalities of muscles can be related to differences in IMF content, metabolism and adipogenic gene expression. Then, potential differences in different muscles in beef cattle were studied. As a second objective, the main sources of experimental variability associated to RT-qPCR results were analyzed following a nested design in order to implement appropriate experimental designs minimizing gene expression variability. To perform the study Longissimus thoracis (LT), Semitendinosus (SM), Masseter (MS), Sternomandibularis (ST) and subcutaneous adipose tissue (SAT) samples of Pirenaica young bulls (n = 4) were collected for IMF, collagen and protein quantification, analysis of adipocyte size distribution and gene expression (PPARG, CEBPA, FAPB4 and WNT10B). A greater IMF content was observed in MS and SM muscles, which had a bimodal adipocyte size distribution while it was unimodal in the muscles LT and ST. This suggest that the different IMF accretion in the muscles studied might be related to different rates of hyperplasia and hypertrophy and that IMF might develop later in LT and ST muscles. The former differences were not mirrored by the expression of the genes analyzed, which might be related to the different contribution of mature and non-mature adipocytes to the total gene expression. When comparing IMF and SAT gene expression, late and early developing tissues respectively, expression of PPARG, CEBPA and FABP4 was higher in the SAT, in agreement with bigger cell size and numbers. The variability study indicates that the analytical factors that add higher variability to the gene expression are the sampling and RT and therefore, it would be appropriate to include those replicates in the design of future experiments. Based on the results, the use of MS and SM muscles could allow less expensive experimental designs and bigger sample size that could permit the detection of lower relevant differences in gene expression.
Open Access
Textile characteristics of fiber from Huacaya alpacas (Vicugna pacos)
(Universidad Nacional de Trujillo (Perú). Facultad de Ciencias Agropecuarias, 2019) Paucar Chanca, Rufino; Alfonso Ruiz, Leopoldo; Soret Lafraya, Beatriz; Mendoza Ordóñez, G.; Alvarado Quezada, F.; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura
Fiber from alpacas represents a substantial component of economic output for South American countries. In this study it determined the textile characteristics of fibers obtained from Huacaya alpacas raised at the South American Camelids Research and Development Center-Lachocc (CRDC-Lachocc) located at The National University of Huancavelica (UNH). Fleece samples were obtained from the mid-side rib area of 74 white alpacas (42 females and 32 males) of varying ages. The Average Fiber Diameter (AFD), Standard Deviation of the Average Fiber Diameter (SDAFD), Fiber Diameter Coefficient of Variation (FDCV), Comfort Factor (CF) and Staple Length (SL) were measured as textile characteristics and related to sex and age group. Most of the fleece samples could be classified as baby alpaca fleece according to the Peruvian Technical Standard classification (231.301.2014). Sex had no influence on any textile characteristic (p > 0.05). Meanwhile, age affected only AFD and CF (p < 0.05). Together the results indicated that alpacas farmed at CRDC-Lachocc had good potential to produce high quality fibers.
Open Access
Preservation of milk in liquid nitrogen during sample collection does not affect the RNA quality for RNA-seq analysis
(BMC, 2025-05-24) Jiménez Montenegro, Lucía; Alfonso Ruiz, Leopoldo; Soret Lafraya, Beatriz; Mendizábal Aizpuru, José Antonio; Urrutia Vera, Olaia; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa; Gobierno de Navarra / Nafarroako Gobernua
Background. Standard procedures for milk sample collection for transcriptome analysis use ice as preservation method, which can afect the RNA stability and requires immediate sample processing. These problems would be eased if the milk samples could be snap-frozen in liquid nitrogen. This study describes the applicability of a new method for milk sample collection and subsequent RNA extraction from milk fat globules, determining whether the quality, integrity and quantity of the RNA extracts met the minimum requirements for downstream RNA-seq. Results. The quality of the extracts measured by A260/280 ratio and the Integrity and Quality (IQ) values obtained fulflled the reference values of 1.9 - 2.1 (P10.05) and failed to meet the RIN≥7 benchmark for RNA-seq (P>0.05). Milk fat globules contain low molecular-weight RNA fragments and minimal 18S and 28S rRNA, suggesting low RIN values were inherent to sample type. Likewise, the RNA concentration from milk fat globules were generally low (120.43±22.27 ng/µL, 102.87±15.64 ng/µL and 109.43±22.69 ng/µL, measured by Nanodrop, Qubit HS and QuanTI Ribogreen, respectively). Nevertheless, RNA-seq yielded 52.7 million paired-end reads per sample. The raw reads passed all quality control parameters having the same sequence-read lengths (151 bp), 100% base-coverage, 49% GC base content, and base quality scores of 36, enabling successful transcriptome profling. Moreover, milk proteins were identifed as the most abundant transcripts in MFG in the analysis of the most expressed genes, indicating that the sequenced reads would accurately refect the transcriptome of this milk fraction. Conclusions. Milk preservation in liquid nitrogen is a suitable sample collection method that overcomes the limitations of immediate sample processing required if ice is used. Thus, this procedure, together with the subsequent RNA isolation from milk fat globules and its sequencing by RNA-seq, would provide a practical and a non-invasive method for measuring the mammary epithelial cell transcriptome, improving the feasibility of conducting studies related to mammary gland and lactation physiology.
Open Access
Comparative description of growth, fat deposition, carcass and meat quality characteristics of Basque and Large White pigs
(EDP Sciences, 2005) Alfonso Ruiz, Leopoldo; Mourot, Jacques; Insausti Barrenetxea, Kizkitza; Mendizábal Aizpuru, José Antonio; Arana Navarro, Ana; Producción Agraria; Nekazaritza Ekoizpena; Gobierno de Navarra / Nafarroako Gobernua
Characteristics of growth, fat deposition, carcass and meat quality of pigs from the Basque Black Pied breed were described and compared with those of Large White pigs. Four pens, two per breed, of eleven pigs born during the same two week period, were simultaneously fattened and slaughtered, under the same conditions. The experiment was carried out over a fixed duration (124 days) and slaughter was carried out at a fixed average age (202 days). Basque pigs showed lower growth and feed efficiency and higher backfat depth (2.6 vs. 1.7 cm, P < 0.001) than Large White pigs. The difference was especially noticeable in the middle subcutaneous fat layer (0.5 cm, P < 0.001). The meat of Basque pigs was darker, redder, more marbled, and with higher pH values than in Large White pigs. Differences in fatty acid composition were observed between breeds but they were not statistically significant (P > 0.05) because of high variability observed between animals. The Basque breed exhibited an early and higher adipose development and a higher activity of enzymes responsible for lipid synthesis than the Large White. The diameter of intramuscular adipose cells was larger in Basque (40.2 vs. 33.0 μm, P < 0.001) than in Large White pigs. The results show the particular characteristics of the Basque breed as compared to pig lines highly selected for lean growth efficiency.
Open Access
The effects of selective breeding against scrapie susceptibility on the genetic variability of the latxa black-faced sheep breed
(EDP Sciences, 2006) Alfonso Ruiz, Leopoldo; Parada Rey, Analia; Legarra, Andrés; Ugarte, Eva; Arana Navarro, Ana; Producción Agraria; Nekazaritza Ekoizpena
Breeding sheep populations for scrapie resistance could result in a loss of genetic variability. In this study, the effect on genetic variability of selection for increasing the ARR allele frequency was estimated in the Latxa breed. Two sources of information were used, pedigree and genetic polymorphisms (fifteen microsatellites). The results based on the genealogical information were conditioned by a low pedigree completeness level that revealed the interest of also using the information provided by the molecular markers. The overall results suggest that no great negative effect on genetic variability can be expected in the short time in the population analysed by selection of only ARR/ARR males. The estimated average relationship of ARR/ARR males with reproductive females was similar to that of all available males whatever its genotype: 0.010 vs. 0.012 for a genealogical relationship and 0.257 vs. 0.296 for molecular coancestry, respectively. However, selection of only ARR/ARR males implied important losses in founder animals (87 percent) and low frequency alleles (30 percent) in the ram population. The evaluation of mild selection strategies against scrapie susceptibility based on the use of some ARR heterozygous males was difficult because the genetic relationships estimated among animals differed when pedigree or molecular information was used, and the use of more molecular markers should be evaluated.
Open Access
Development of a duplex qPCR assay with locked nucleic acid probes for A, B and E kappa-casein variants detection
(Springer Nature, 2022) Jiménez Montenegro, Lucía; Mendizábal Aizpuru, José Antonio; Alfonso Ruiz, Leopoldo; Azparren Domínguez, Leire; Urrutia Vera, Olaia; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD
Milk proteins determine important milk technological characteristics. Among caseins, Ƙ-casein has been correlated with fat and protein content and cheese yield. Fourteen Ƙ-caseins variants have been described but the alleles A, B and E are the most important ones due to their frequency and/or influence on the technological aptitudes of milk. Therefore, in the present study two different duplex qPCR assays with locked nucleic acid probes (for positions 13104 and 13124 of the Ƙ-casein gene) were developed for the detection of A, B and E variants. Firstly, DNA isolation method from milk somatic cells and hair was optimised. The developed 13124-qPCR assay showed an increased sensitivity reaching up to 6.7 copies DNA copies/reaction at a 95% confidence level with A, B and E alleles reference samples. The 13104-qPCR assay reached up to 6.7 DNA copies/reaction for A allele reference sample and 67 DNA copies/reaction for B and E samples. Intra-assay variation results were below 6%. Applicability was determined using DNA samples from animals with known genotype for Ƙ-casein (AA, AB, BB, BE, AE, EE) and both assays were able to discriminate among the six genotypes with 100% accuracy. Thus, this qPCR method represents a sensitive and rapid option for the detection of Ƙ-casein alleles in both hair and milk samples.
Open Access
Adipose tissue modification through feeding strategies and their implication on adipogenesis and adipose tissue metabolism in ruminants
(MDPI, 2020) Urrutia Vera, Olaia; Mendizábal Aizpuru, José Antonio; Alfonso Ruiz, Leopoldo; Soret Lafraya, Beatriz; Insausti Barrenetxea, Kizkitza; Arana Navarro, Ana; Agronomia, Bioteknologia eta Elikadura; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD; Agronomía, Biotecnología y Alimentación; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
Dietary recommendations by health authorities have been advising of the importance of diminishing saturated fatty acids (SFA) consumption and replacing them by polyunsaturated fatty acids (PUFA), particularly omega-3. Therefore, there have been efforts to enhance food fatty acid profiles, helping them to meet human nutritional recommendations. Ruminant meat is the major dietary conjugated linoleic acid (CLA) source, but it also contains SFA at relatively high proportions, deriving from ruminal biohydrogenation of PUFA. Additionally, lipid metabolism in ruminants may differ from other species. Recent research has aimed to modify the fatty acid profile of meat, and other animal products. This review summarizes dietary strategies based on the n-3 PUFA supplementation of ruminant diets and their effects on meat fatty acid composition. Additionally, the role of n-3 PUFA in adipose tissue (AT) development and in the expression of key genes involved in adipogenesis and lipid metabolism is discussed. It has been demonstrated that linseed supplementation leads to an increase in alpha-linolenic acid (ALA) and eicosapentaenoic acid (EPA), but not in docosahexaenoic acid (DHA), whilst fish oil and algae increase DHA content. Dietary PUFA can alter AT adiposity and modulate lipid metabolism genes expression, although further research is required to clarify the underlying mechanism.
Open Access
Impact of incorporating greenhouse gas emission intensities in selection indexes for sow productivity traits
(Elsevier, 2019) Alfonso Ruiz, Leopoldo; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD
Genetic improvement programmes should incorporate emerging challenges about environmental concerns into breeding goals. The large volume of pig meat production implies important greenhouse gas (GHG) emissions despite its lower carbon footprint per animal in front of ruminant productions. The different breeding goals considered by swine industry depending on different purebred lines, or line crosses adapted to different market demands and production constraints, could mask the effect of incorporating GHG emissions into selection indexes for improving sow productivity traits in nucleus populations. This paper analysed this effect following a methodological approach consisting in augmenting existing selection indexes derived from profit functions. An index previously described in the literature including litter size at birth, piglet perinatal survival, piglet survival to weaning, age at first conception and weaning to conception interval, was employed. This index was expanded to include GHG emissions calculating the emission intensities per litter, assuming a finished pig market and different scenarios and financial costs of GHG emissions. Results indicated that the inclusion of GHG emissions diminished the economic weight of litter size and piglet survival vs. the age at first conception and the interval weaning to conception, but did not affect significantly the contributions of these traits in the selection indexes. The improvement of sow productivity traits diluted relevantly the GHG emissions per piglet produced, and so, per kg of pork produced. The approach used in this study, despite its limitations in front of bio-economic models, has shown to be a simple and flexible way to analyse the effect of incorporating GHG emissions into existing selection indexes.