Person: Alfonso Ruiz, Leopoldo
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Alfonso Ruiz
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Leopoldo
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Agronomía, Biotecnología y Alimentación
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IS-FOOD. Research Institute on Innovation & Sustainable Development in Food Chain
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0000-0002-5662-9997
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1974
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Publication 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 ElikaduraFiber 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.Publication Open Access DNA extraction procedures and validation parameters of a real-time PCR method to control milk containing only A2 β-casein(Elsevier, 2022) Jiménez Montenegro, Lucía; Mendizábal Aizpuru, José Antonio; Alfonso Ruiz, Leopoldo; Institute on Innovation and Sustainable Development in Food Chain - ISFOOD; Gobierno de Navarra / Nafarroako Gobernua; Universidad Pública de Navarra / Nafarroako Unibertsitate PublikoaBovine milk mainly contains two types of β-casein: A1 and A2 variants. In recent years, a new variety of cows’ milk has emerged in the dairy sector called “A2 milk”. This novel product is characterised by the absence of A1 β-casein, which has been associated with possible gastrointestinal discomfort due to β-casomorphin-7 (BCM-7) release during gastrointestinal digestion. In this context, methods to verify the A1 allele absence in A2 milk are required as a quality control in the A2 milk commercialisation. Therefore, the aim of the present study was to develop a locked nucleic acid (LNA) probe-based duplex real-time PCR (qPCR) assay for A1 allele detection in A2 milk samples. Firstly, four DNA isolation methods from milk somatic cells were optimised and evaluated. The results suggests that the commercial kit NucleoSpin Tissue was the most suitable method in terms of DNA quality and amplificability for downstream applications. Then, optimisation and validation of the qPCR assay were carried out. For both A1 and A2 alleles, the absolute limits of detection of this qPCR assay were 7.3 DNA copies/reaction (2 x 10−5 ng DNA) and 30.4 DNA copies/reaction (0.1 ng DNA) at a 95% confidence level with synthetic reference DNA samples and heterozygous genotyped DNA sample, respectively. The relative limits of detection were 2% (15 copies) and 5% (152 copies) for the A1 allele in A2 samples at 95% confidence with synthetic reference and genotyped DNA samples, respectively. The qPCR assay was robust, with intra- and inter-assay variability below 4.3%, and specific, differentiating between A1 and A2 alleles with 100% genotyping accuracy. In conclusion, this cost-effective and fast method could be used to discriminate A1 allele in A2 samples and, consequently, to verify the A1 allele absence in “A2 milk” by screening commercial products on the market.Publication Open Access Análisis de la relación entre el espesor de la grasa dorsal y el tamaño de los adipocitos en cerdas reproductoras(Asociación Interprofesional para el Desarrollo Agrario, 2007) Mendizábal Aizpuru, José Antonio; Abadía Durán, Silvia; Abaurrea Aramburu, Eneko; Alfonso Ruiz, Leopoldo; Producción Agraria; Nekazaritza EkoizpenaLa correcta gestión de las reservas grasas durante el ciclo productivo de las cerdas reproductoras es un factor clave para la obtención de unos buenos resultados productivos. El principal depósito graso en las cerdas lo constituye el tejido graso subcutáneo, por lo que la medición del espesor graso dorsal es un buen indicador del estado de las reservas corporales. Aunque existen diversos puntos para la medición del espesor, se recomienda hacerlo a nivel de la última costilla flotante, punto que se conoce como P2 (Quiniou, 2004). En esta localización se pueden distinguir hasta tres capas distintas de grasa subcutánea (Moody y Zobrisky, 1966). Distintos trabajos han observado un desarrollo diferencial de las tres capas durante el crecimiento de los animales (Alfonso, 2004; Fortin, 1986), así como diferencias en su actividad lipogénica (Cámara et al., 1996). Por tanto, el estudio de las características de cada capa considerada como distinto depósito graso puede ser importante para conocer los mecanismos de acumulación y movilización de las reservas grasas. En este sentido, en el presente trabajo se estudia la relación entre el espesor de la grasa y el tamaño de los adipocitos de cada una de las capas descritas en la grasa subcutánea dorsal.Publication Open Access Ayuda a las decisiones de apareamiento en poblaciones animales con información genealógica incompleta(Asociación Interprofesional para el Desarrollo Agrario, 2018) Alfonso Ruiz, Leopoldo; Institute on Innovation and Sustainable Development in Food Chain - ISFOODLos coeficientes de parentesco y de relación media de parentesco son dos herramientas útiles para determinar apareamientos con el objetivo de controlar, de forma práctica, el aumento de la consanguinidad en poblaciones animales. No obstante, su utilidad se ve condicionada por la cantidad de información genealógica disponible, pues ésta determina, en ausencia de información genómica, la precisión con que podemos estimarlos. Cuando la información genealógica sea incompleta, afectando de forma distinta a los animales, la precisión de las estimaciones de los coeficientes variará entre animales, comprometiendo el resultado de las decisiones de apareamiento. El objetivo del trabajo es calcular algunas medidas de la fiabilidad de la estimación de los coeficientes de parentesco y de relación media de parentesco. Se plantea la utilización de dos medidas basadas en la proporción de antecesores conocidos en las cinco generaciones anteriores de un animal y en la incorporación de información conocida, pero no contenida en la genealogía, a través del concepto de grupos genéticos. Los resultados muestran que estas medidas pueden ayudar a mejorar las decisiones de elección de reproductores y de acoplamiento.Publication 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 EkoizpenaBreeding 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.Publication 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 - ISFOODGenetic 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.Publication 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 EkoizpenaAnatomical 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.Publication 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 AlimentaciónAdipogenesis, 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.Publication Open Access Conversión de las explotaciones de vacuno de leche a la producción de leche A2 ante una posible demanda del mercado: posibilidades e implicaciones(Asociación Interprofesional para el Desarrollo Agrario, 2019) Alfonso Ruiz, Leopoldo; Urrutia Vera, Olaia; Mendizábal Aizpuru, José Antonio; Institute on Innovation and Sustainable Development in Food Chain - ISFOODEl mercado de la leche de vaca, tras la desaparición de las cuotas lácteas, está provocando una importante restructuración en el sector. Una de las alternativas que están desarrollando, fundamentalmente las pequeñas explotaciones familiares, es la diferenciación de sus productos. En ese contexto, ha aparecido un tipo diferenciado de leche conocida como leche A2. Esta se caracteriza por estar libre de la variante A1 de la proteína β-caseína que, debido a una mayor producción de β-casomorfina-7 tras la digestión, puede dar lugar a intolerancias y afecciones gastrointestinales. En este trabajo se analizan las posibilidades e implicaciones de una selección a favor de la β-caseína A2 en el caso concreto de las explotaciones familiares de Guipúzcoa. Los resultados obtenidos a partir de 1868 hembras genotipadas muestran que la frecuencia del alelo que codifica la proteína A2 es 0,55, y no se han encontrado asociaciones desfavorables entre el alelo A2 y el valor genético para caracteres productivos, funcionales y morfológicos, ni con otros genes ni haplotipos. Bajo esta situación de partida, y empleando exclusivamente toros A2A2, la utilización o no de semen sexado, junto con las diferentes ratios de reposición que se apliquen, determinarán el periodo y el coste económico de la conversión de los rebaños a la producción de leche A2. En un escenario de costes razonables esta se estima entre los 8 y los 15 años, pudiendo llegar a suponer a una explotación de 150 vacas unos costes de unos 6.000 y 1.000 €/año, respectivamente.Publication 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 GobernuaCharacteristics 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.