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
Efecto de la utilización de pienso enriquecido en ácidos grasos poliinsaturados en corderos en crecimiento sobre el desarrollo, la composición de la grasa de la carne y la expresión de los principales genes lipógenos del tejido adiposo
(2016) Urrutia Vera, Olaia; Arana Navarro, Ana; Soret Lafraya, Beatriz; Producción Agraria; Nekazaritza Ekoizpena
En los últimos años, el aporte de ácidos grasos poliinsaturados (PUFA) de tipo n-3 en la alimentación animal es una vía que está siendo estudiada para mejorar el perfil lipídico de la carne, debido a que es un alimento con una proporción considerable de ácidos grasos saturados que se asocian a ciertas enfermedades crónicas y además existe un interés creciente por consumir alimentos más saludables. Sin embargo, algunos estudios muestran que el aumento de ácidos grasos saludables como el ácido a-linolénico (ALA), el ácido eicosapentaenoico (EPA), el ácido docosahexaenoico (DHA) y el isómero C18:2 c9t11 del ácido linoleico conjugado (CLA) en la carne tras la adición de fuentes de PUFA en las dietas de los animales no es muy significativo. Las hipótesis planteadas para esta discordancia apuntan a la regulación de la expresión de genes implicados en el metabolismo lipídico. Por lo tanto, el objetivo del presente estudio ha sido valorar el efecto de la incorporación materias primas ricas en PUFA (semilla de lino y chía ricas en ALA y microalgas marinas con un alto contenido en DHA) en la alimentación de corderos de raza Navarra durante el cebo de sobre las características de crecimiento y de la canal, el desarrollo del tejido graso y la composición en ácidos grasos de la carne y de la grasa, además de analizar varios parámetros relacionados con la calidad de la carne y valorar su aceptabilidad por parte de los consumidores. Por último, con objeto de profundizar en la comprensión de los mecanismos moleculares por medio de los cuales los PUFA pueden ejercer su acción en el tejido adiposo, se ha estudiado la expresión de varios genes lipogénicos y adipogénicos.
Open Access
Insights into the role of major bioactive dietary nutrients in lamb meat quality: a review
(BMC, 2022) Álvarez-Rodríguez, Javier; Urrutia Vera, Olaia; Lobón, Sandra; Ripoll, Guillermo; Bertolín, Juan R.; Joy Torrens, Margalida; Agronomía, Biotecnología y Alimentación; Agronomia, Bioteknologia eta Elikadura
Feed supplementation with α-linolenic acid (ALA) and linoleic acid (LA) increases their content in muscle, ALA increases n-3 polyunsaturated fatty acids and decrease n-6/n-3 ratio in muscle, and LA increases rumenic acid. However, high LA supplementation may have negative effects on lambs’ lipid oxidative stability of meat. When the sources of ALA and LA are fed as fresh forage, the negative effects are counterbalanced by the presence of other bioactive compounds, as vitamin E (mainly α-tocopherol) and polyphenols, which delay the lipid oxidation in meat. There is a wide consensus on the capability of vitamin E delaying lipid oxidation on lamb meat, and its feed content should be adjusted to the length of supplementation. A high dietary inclusion of proanthocyanidins, phenolic compounds and terpenes reduce the lipid oxidation in muscle and may improve the shelf life of meat, probably as a result of a combined effect with dietary vitamin E. However, the recommended dietary inclusion levels depend on the polyphenol type and concentration and antioxidant capacity of the feedstuffs, which cannot be compared easily because no routine analytical grading methods are yet available. Unless phenolic compounds content in dietary ingredients/supplements for lambs are reported, no specific association with animal physiology responses may be established.
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
Predicting beef carcass fatness using an image analysis system
(MDPI, 2021) Mendizábal Aizpuru, José Antonio; Ripoll, Guillermo; Urrutia Vera, Olaia; Insausti Barrenetxea, Kizkitza; Soret Lafraya, Beatriz; Arana Navarro, Ana; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD
The amount and distribution of subcutaneous fat is an important factor affecting beef carcass quality. The degree of fatness is determined by visual assessments scored on a scale of five fatness levels (the SEUROP system). New technologies such as the image analysis method have been developed and applied in an effort to enhance the accuracy and objectivity of this classification system. In this study, 50 young bulls were slaughtered (570 ± 52.5 kg) and after slaughter the carcasses were weighed (360 ± 33.1 kg) and a SEUROP system fatness score assigned. A digital picture of the outer surface of the left side of the carcass was taken and the area of fat cover (fat area) was measured using an image analysis system. Commercial cutting of the carcasses was performed 24 h post-mortem. The fat trimmed away on cutting (cutting fat) was weighed. A regression analysis was carried out for the carcass cutting fat (y-axis) on the carcass fat area (x-axis) to establish the accuracy of the image analysis system. A greater accuracy was obtained by the image analysis (R2 = 0.72; p < 0.001) than from the visual fatness scores (R2 = 0.66; p > 0.001). These results show the image analysis to be more accurate than the visual assessment system for predicting beef carcass fatness.
Open Access
Efecto de la dieta en el nivel de expresión génica de las enzimas Acetil-Coa Carboxilasa, Lipoprotein Lipasa y Estearoil-Coa Desaturasa, enzimas lipogénicas implicadas en la síntesis y acumulación de ácidos grasos insaturados en el depósito subcutáneo de ovino
(2010) Urrutia Vera, Olaia; Arana Navarro, Ana; Escuela Técnica Superior de Ingenieros Agrónomos; Nekazaritza Ingeniarien Goi Mailako Eskola Teknikoa; Producción Agraria; Nekazaritza Ekoizpena
El objetivo del trabajo fin de carrera realizado es el de estudiar el efecto de la dieta en el nivel de expresión génica de varias enzimas lipogénicas implicadas en la síntesis de ácidos grasos insaturados en el deposito subcutáneo de ovino. El trabajo se ha realizado utilizando 36 corderos ternascos, provenientes de la explotación experimental de ovino de carne “El Serrón” del ITGG en Valtierra, divididos en tres lotes de 12 animales que recibieron tres dietas diferentes en función de su composición en lino; lote control (0% lino), lote 5% (5% de lino) y lote 10% (10% de lino). Los corderos presentaron diferencias en la composición de ácidos grasos del tejido subcutáneo como consecuencia de la inclusión del lino y aquellos que ingirieron lino en la racion presentaron mayor cantidad en la composición de ácido linolénico, ácido vaccénico y del C18:2n6t9t12 y el total de ácidos grasos omega 3. Los genes, cuya expresión génica se ha analizado, han sido: la Acetil-CoA Carboxilasa (ACC), Lipoprotein Lipasa (LPL) y la Estearoil-CoA Desaturasa (SCoAD). La metodología a utilizar consiste en la cuantificación de la expresión génica de las enzimas Acetil-CoA Carboxilasa (ACC), Lipoprotein Lipasa (LPL) y Estearoil-CoA Desaturasa (SCD) mediante la técnica de PCR a tiempo real utilizando el método comparativo de Ct’s, siendo la β actina el gen endógeno. Los resultados de este trabajo indican que la inclusión de lino en un 10% en la dieta de los corderos disminuye la expresión génica de la enzima relacionada con la síntesis de novo de ácidos grasos, la Acetil-CoA Carboxilasa. Además, el lino en la dieta, tanto al 5% como al 10%, incrementa la expresión génica de la enzima Lipoprotein Lipasa, enzima encargada de la captación desde la sangre de los ácidos grasos provenientes de la dieta. Por último, los corderos que ingirieron lino al 10% vieron disminuida la expresión génica de la enzima Estearoil-CoA Desaturasa. Teniendo en cuenta estos resultados, podría indicar que las dietas ricas en ácidos grasos poliinsaturados de cadena larga además de aumentar la incorporación de ácidos grasos provenientes de la alimentación, disminuyen la síntesis de novo y la desaturación. Por lo que sería conveniente tener en cuenta estos efectos en el momento de diseñar las dietas a suministrar a los corderos con objeto de modificar determinados ácidos grasos en la carne y grasa de éstos.
Open Access
Detection of minced lamb and beef fraud using NIR spectroscopy
(Elsevier, 2019) López Maestresalas, Ainara; Insausti Barrenetxea, Kizkitza; Jarén Ceballos, Carmen; Pérez Roncal, Claudia; Urrutia Vera, Olaia; Beriain Apesteguía, María José; Arazuri Garín, Silvia; Ingeniaritza; Agronomia, Bioteknologia eta Elikadura; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD; Ingeniería; Agronomía, Biotecnología y Alimentación; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
The aim of this work was to investigate the feasibility of near-infrared spectroscopy (NIRS), combined with chemometric techniques, to detect fraud in minced lamb and beef mixed with other types of meats. For this, 40 samples of pure lamb and 30 samples of pure beef along with 160 samples of mixed lamb and 156 samples of mixed beef at different levels: 1-2-5-10% (w/w) were prepared and analyzed. Spectral data were pre-processed using different techniques and explored by a Principal Component Analysis (PCA) to find out differences among pure and mixed samples. Moreover, a PLS-DA was carried out for each type of meat mixture. Classification results between 78.95 and 100% were achieved for the validation sets. Better rates of classification were obtained for samples mixed with pork meat, meat of Lidia breed cattle and foal meat than for samples mixed with chicken in both lamb and beef. Additionally, the obtained results showed that this technology could be used for detection of minced beef fraud with meat of Lidia breed cattle and foal in a percentage equal or higher than 2 and 1%, respectively. Therefore, this study shows the potential of NIRS combined with PLS-DA to detect fraud in minced lamb and beef.
Open Access
Improving the short‑term efficiency of rock phosphate‑based fertilizers in pastures by using edaphic biostimulants
(Springer, 2016) Fernández, L.; Baigorri, R.; Urrutia Vera, Olaia; Erro, J.; Aparicio Tejo, Pedro María; Yvin, J. C.; García Mina, José M.; IdAB. Instituto de Agrobiotecnología / Agrobioteknologiako Institutua; Gobierno de Navarra / Nafarroako Gobernua
Background: The use of reactive rock phosphate (RP) in acidic soils as a phosphate (P) source for pastures and crops presents attractive economic advantages with respect to soluble phosphate. However, some studies have demonstrated that the short-term (1-year) efficiency of RP, compared with that of water-soluble P, is relatively poor. This fact penalizes not only the yield and quality of the earlier harvests, but even the whole final yield when the crop is affected by some abiotic or biotic stress at the beginning of the cycle. In the present study, we investigated the ability of new edaphic biostimulants to increase the short-term efficiency of RP-based fertilizer as a P source for pastures cultivated in acid soils. To this end, we have granulated rock phosphate with two edaphic biostimulants: tryptophan and a heteromolecular organic complex formed by humic acid and tryptophan through iron bridges, and compared their short-term P fertilizer efficacy with that of single superphosphate and rock phosphate. Results: Soil incubation studies showed that the heteromolecular complex humic acid–tryptophan and Tryptophan were able to increase soil CO2 production compared with native soil, rock phosphate, and superphosphate. Likewise, the presence of humic acid–tryptophan complex and Trp significantly increases plant-available phosphate compared with rock phosphate, up to levels similar to those of superphosphate. Plant (ray grass)–soil–pot studies showed that rock phosphate/(humic acid–tryptophan) formulation yielded values for both ray grass dry matter production and shoot P concentration, clearly higher than those of rock phosphate and rock phosphate/tryptophan. In addition, the results associated with rock phosphate/(humic acid–tryptophan) were similar to those for superphosphate, after 3 months of cultivation. Conclusions: Taken together, these results showed the suitability of the use of specific humic acid-based edaphic biostimulants to improve the short-term effect of rock phosphate fertilizers as a phosphate source for pastures cultivated in acid soils.
Open Access
Efecto de una alimentación rica en semilla de lino y microalgas con alto contenido en ácidos grasos poliinsaturados en la expresión génica de enzimas lipogénicas en la grasa subcutánea e intramuscular de la carne de corderos
(2011) Urrutia Vera, Olaia; Arana Navarro, Ana; Escuela Técnica Superior de Ingenieros Agrónomos; Nekazaritza Ingeniarien Goi Mailako Eskola Teknikoa
Entre las causas de la disminución del consumo actual de carne de cordero se encuentra su alto contenido en ácidos grasos saturados ya que existe una preocupación por la salud y este tipo de grasas suponen un mayor riesgo de padecer obesidad y enfermedades cardiovasculares (Baghurst, 2004; Givens, 2005). Actualmente, se utilizan estrategias de alimentación para los rumiantes utilizando materias primas ricas en ácidos grasos (AGs) del tipo omega 3, con el objeto de modificar la composición de los AGs de su carne. Estudios previos han demostrado que este tipo de alimentación rica en ácidos grasos poliinsaturados (PUFAs) aumenta la composición de estos AGs en el músculo pero inhibe la actividad de algunas enzimas implicadas en la síntesis endógena y acumulación de los PUFAs (Ntambi, 1992; Bernard et al., 2005) no obteniendo el enriquecimiento esperado en estos AGs. En este trabajo se ha profundizado en el estudio de la expresión de genes en el músculo y tejido adiposo implicados en el metabolismo de los AGs en corderos alimentados con dietas ricas en PUFAs como el lino, con alto contenido en ácido alfalinolénico (ALA) y microalgas, abundante en ácido docosahexaenoico (DHA) y ácido eicosapentaenoico (EPA). Se utilizaron 33 corderos de Raza Navarra divididos en tres lotes cuya alimentación difería en el contenido y tipo de PUFAs: lote C (control), lote L suplementado con un 10% de lino y lote LA con 5% de lino y 3,75% de microalgas. Mediante RT-PCR a tiempo real se cuantificó la expresión de los genes ACC, LPL y SCoAD. Los resultados muestran diferentes patrones en la expresión de dichos genes en función del depósito. En el tejido adiposo intramuscular, la incorporación de lino y lino y las algas provocaron un descenso en la expresión génica de la ACC y SCoAD y un aumento en la expresión de la LPL respecto al grupo control. En el tejido adiposo subcutáneo, la expresión de la ACC y SCoAD aumentaron y disminuyó la expresión de la LPL en el lote L respecto al control, sin embargo, la inclusión de lino y las algas provocó un aumento en la expresión de la ACC, una disminución en la SCoAD y la LPL no presentó variación respecto al grupo control.
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.