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.

Bovine milk mainly contains two types of beta-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 beta-casein, which has been associated with possible gastrointestinal discomfort due to ss beta-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.

Nowadays, the meat industry requires non-destructive, sustainable, and rapid methods that can provide objective and accurate quality assessment with little human intervention. Therefore, the present research aimed to create a model that can classify beef samples from longissimus thoracis muscle according to their tenderness degree based on hyperspectral imaging (HSI). In order to obtain different textures, two main strategies were used: (a) aging type (wet and dry aging with or without starters) and (b) aging times (0, 7, 13, 21, and 27 days). Categorization into two groups was carried out for further chemometric analysis, encompassing group 1 (ngroup1 = 30) with samples with WBSF ˂ 53 N whereas group 2 (ngroup2 = 28) comprised samples with WBSF values ≥ 53 N. Then, classification models were created by applying the partial least squares discriminant analysis (PLS-DA) method. The best results were achieved by combining the following pre-processing algorithms: 1st derivative + mean center, reaching 70.83% of correctly classified (CC) samples and 67.14% for cross validation (CV) and prediction, respectively. In general, it can be concluded that HSI technology combined with chemometrics has the potential to differentiate and classify meat samples according to their textural characteristics.

The protein fraction of β-casein may play a key role in the manifestation of a new intolerance: milk protein intolerance. The most common forms of β-casein among dairy cattle breeds are A1 and A2 β-casein. During gastrointestinal digestion of A1 β-casein, an opioid called peptide β-casomorphin-7 (BCM-7) is more frequently released, which can lead to adverse health outcomes. For that reason, novel products labelled as “A2 milk” or “A1-free dairy products” have appeared on the market. In this context, a bibliometric analysis on A2 β-casein research was carried out through the Web of Science (WoS) database. The main objective of this work was to provide an overview of the state of the art in the field of β-casein A2 by analyzing the number of publications per year, trends in thematic content, the most frequently used terms, and the most important institutions and countries in the field. This bibliometric study showed that a greater effort is needed to determine the possible implications of this novel product for human health and the market.

This study aimed to assess the influence of ageing on the volatile compounds, as well as odour and flavour attributes of lamb meat from the Navarra breed. Twenty-one male lambs were fed a commercial concentrate diet after weaning and were harvested at 101 ± 6.5 days of age. From the Longissimus thoracis, 26 volatile compounds were identified, with hexanal, 2-propanone, and nonanal the most abundant (57.17% relative percentage abundance, RPA). The effect of ageing (1 vs. 4 d) was observed (p < 0.05) in six compounds: 1,4-dimethylbenzene decreased with ageing, while tridecane, 3-methylbutanal, 2-heptanone, 3-octanone, and 1-octen-3-ol increased. In general, ageing was linked to a decrease in livery and bloody flavour, bloody odour and ethanal, and an increase in pentane, hexanal, and heptanal, which are usually associated with fresh green grass and fat descriptors. Consequently, ageing lamb from the Navarra breed for four days might have a positive effect on meat sensory odour and flavour quality.

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.

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.

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 α-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.

BOOK REVIEWS - CRITICA DE LIBROS - CRITICA DE LIVROS Patrick Doyle: Civilising Rural Ireland: The Co-Operative Movement, Development and the Nation-State, 1889-1939 Tony Varley Ana Cristina Roque, Cristina Brito y Cecilia Veracini (Eds.): Peoples, Nature and Environments: Learning to Live Together Antonio Ortega Santos Sylvie Bépoix y Hervé Richard (Dirs.): La forêt au Moyen Âge Juan José Larrea Manuel González de Molina, David Soto, Gloria Guzmán Casado, Juan Infante, Eduardo Aguilera, Jaime Vila y Roberto García Ruíz: Historia de la Agricultura Española desde una perspectiva biofísica, 1900-2010 Ernesto Clar Enric Saguer (Ed.): Els cortals empordanesos, del segle xiii al xxi. Sis estudis Antònia Morey Tous Enric Vicedo (Ed.): Recs històrics: Pagesia, història i patrimonio Mar Ortega-Reig Francisco Acosta (Coord.): La aurora de rojos dedos: El Trienio Bolchevique desde el sur de España Antonio Herrera González de Molina Antonio Luis López Martínez: Las ferias de ganado en Andalucía Occidental y Extremadura, 1830-1958 José Antonio Mendizabal

ObjectivesThe objective of the study was to assess the evolution of the bloom color in beef aged for 8 d to establish the moment of measurement in which values of L*, a* and b* stabilize and are representative of the characteristic color of the meat from beef fed different diets.Materials and MethodsIn the current study, eight young Pirenaica bulls were used. The bulls were born and reared on a private commercial Protected Denomination Origin (PDO)-approved farm located in the region of Navarra (Northern Spain). After weaning at approximately 4 mo of age, the calves were administered the same diet until month 12. The bulls were separated in two groups and each of group was fed a different energy level diet (High energy, H: 2914.2 kcal/kg vs. Low energy, L: 2548.4kcal/kg) until slaughter at 18 mo of age. Diet was based on barley (H: 26% vs. L: 22%), corn (H: 50% vs. L: 45%) and soja (H: 17% and L: 17%). The research was conducted under the highest standards of humane care and use of animals in accordance with European guidelines (EU). Longissimus dorsi muscle was removed after 24 h. post-mortem from the left side of the carcasses, pH was measured, and the meat was transported to the Meat Science Laboratory at the Public University of Navarra (Pamplona, Spain) under refrigeration. Steaks were aged in vacuum for 8 d post-mortem, which is the typical period for this type of meat under the PDO Ternera de Navarra. After aging, L*, a*, and b*were recorded every 3 min (5 repetitions per sample) for 102 min with a Minolta CM 2002 Spectrophotocolorimeter. Data were analyzed using the Linear General Model procedure with the IBM SPSS Statistics 24, and significance was determined at P < 0.05.ResultsThe pH values were 5.56 (H) and 5.50 (L) (P < 0.05) thus, no DFD meat was observed. Color differed depending on diet (L*H: 28.88 vs. L*L: 34.26, P < 0.01; a*H: 26.33 vs. a*L: 18.11, P < 0.001; b*H: 11.58 vs. b*L: 7.94, P < 0.001) even if the initial pigment content was not statistically different (H: 5.34 mg/g vs. L: 4.74 g/g; P = 0.107). In fact, beef from the H diet showed higher a* and b* values, and lower L* values than beef from the L diet (P < 0.05). Nonetheless, the time of stabilization for a practical color measurement did not differ between diets.ConclusionIn conclusion, despite the effect of diet on the initial beef color differences, the results of the current study showed that 15 min of meat exposure to oxygen is the minimum in either cases prior to taking measurements of color on beef aged 8 d.

ResumenEl 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.

Bovine dairy market, after the abolition of milk quotas, is causing dairy sector restructuration. Product differentiation is one of the strategies that are implementing dairy farms, mainly small family farms. In this context, a new type of differentiated cow s milk has appeared known commercially as A2 milk. This type of milk is devoid of A1 beta-casein protein, which can cause intolerance and gastrointestinal discomfort due to higher beta-casomorphin-7 release after digestion. In the present work, the possibilities and implications of a selection in favor of A2 beta-casein were analysed in family farms in Guipuzcoa (Spain). The results obtained from 1868 genotyped females showed that the frequency of allele coding the A2 protein was 0.55 and no unfavorable associations were found between A2 allele and the breeding values for production, functional and morphological traits nor other genes or haplotypes. Under this scenario, and using only A2A2 bulls, the use or not of sexed semen, together with different replacement rates that are applied, will determine the period and the economic cost of herd conversion to A2 milk production. In a reasonable cost strategy, the estimated conversion period is between 8 and 15 years, which, in a 150-cow farm, could imply an approximate cost between 6,000 and 1,000 (sic)/year, respectively.

Cellularity of adipose tissue in domesticated animals varies not only with species, sex, age and management conditions but also with depot. Differences in depots are important in animal production because of the economic and welfare implications and in humans in relation to obesity. The final amount of fat and its composition depends on the differentiation of mesenchymal multipotent precursor cells into mature adipocytes (adipogenesis) capable of fatty acid and triglyceride synthesis (lipogenesis), both processes being regulated by different key adipogenic and lipogenic genes, some of are well known and have been described. Histologically, differences can be classified as hyperplasia (an increase in adipocyte number) and hypertrophy (an increase in adipocyte size), processes that can produce adipocyte size distributions that are not necessarily Gaussian. A detailed description of the type of adipocyte size distribution can help distinguish the different adipocyte populations within depots and characterise each not only in terms of the size but also the number of the constituting cells. This description can help better understand the development and role of the different depots. It can also help when analysing causal relationships with adipogenic drivers and lipogenic enzymes involved in lipid metabolism.

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.

The ability to accumulate intramuscular fat (IMF) is a highly variable characteristic in beef cattle. In breeds with a low tendency to accumulate IMF, this can lead to compromised meat quality because of the contribution of fat to such organoleptic attributes as juiciness and taste. This study considered adiposity and gene expression of some of the main markers involved in adipogenesis and lipid metabolism in the subcutaneous (SC) adipose tissue (AT) and the longissimus thoracis muscle (LM) and investigated differences in adipogenic regulation between the tissues during growth and fattening under different conditions. Pirenaica beef cattle were chosen for the study due to the breed's low tendency to accumulate IMF and the breed's regional importance. The young Pirenaica bulls used (n=16) were allocated to four groups and slaughtered at 6, 12 and 18 months. From 12 months onwards the bulls slaughtered at 18 months were fed diets having different energy densities. Backfat thickness increased from 6 to 12 months (P<0.05) but then was unchanged, while other fattening parameters such as percentage chemical fat and marbling did not vary. The adipose cell size distribution displayed a bimodal distribution for SC adipocytes and a unimodal distribution for IMF cells, suggestive of tissue-specific hyperplasia. Gene expression of peroxisome proliferator-activated receptor γ (PPARG), CCAAT/enhancer-binding protein α (CEBPA), sterol regulatory element-binding transcription factor 1 (SREBF1), wingless-type MMTV integration site family 10B (WNT10B), fatty acid-binding protein 4 (FABP4), acetyl Co-A carboxylase α, lipoprotein lipase and fatty acid synthase (FASN) were determined by real-time quantitative PCR. Expression did not differ between the experimental groups within the tissues but did differ between the tissues: PPARG, FABP4 and FASN were upregulated in the SC AT, while CEBPA, WNT10B and SREBF1 were upregulated in the LM. Although age and diet energy density did not have a significant effect on increasing the amount of IMF, these factors could have influenced adipocyte development in this tissue differently than in the SC AT. This was evidenced by the different size distributions of the cells in the two tissues, and the differing expression patterns of certain markers in the SC AT and the LM, which may indicate a differential role of PPARG and WNT10B in triggering adipocyte proliferation and fat accumulation capacity.

ResumenEl tamaño de los adipocitos, principales células constituyentes del tejido adiposo, ha sido objeto de numerosos estudios por el hecho de estar relacionado con el grado de desarrollo y la actividad metabólica de ese tejido.Su análisis resulta en ocasiones complejo dada la bimodalidad de su distribución.Este trabajo presenta un método de análisis basado en contrastar, en primer lugar la unimodalidad de la distribución del tamaño de los adipocitos, frente a una distribución bimodal.Posteriormente propone algunos parámetros para describir adecuadamente la distribución bimodal.Para su aplicación se desa rrolló un sencillo programa informático que se utiliza, a modo de ejemplo, para analizar los datos de dos trabajos, previamente publicados, sobre el desarrollo adipocitario.Los resultados muestran el interés de utilizar contrastes de bimodalidad frente a la mera inspección visual de los histogramas de distribución.Además, cuando la hipótesis de unimodalidad es rechazada, la utilización de parámetros descriptivos de bimodalidad, como el porcentaje de adipocitos por encima del punto de inflexión entre

This study examined the effect of linseed and algae on growth and carcass parameters, adipocyte cellularity, fatty acid profile and meat quality and gene expression in subcutaneous and intramuscular adipose tissues (AT) in lambs. After weaning, 33 lambs were fed three diets up to 26.7 ± 0.3 kg: Control diet (barley and soybean); L diet (barley, soybean and 10% linseed) and L-A diet (barley, soybean, 5% linseed and 3.89% algae). Lambs fed L-A diet showed lower average daily gain and greater slaughter age compared to Control and L (P < 0.001). Carcass traits were not affected by L and L-A diets, but a trend towards greater adipocyte diameter was observed in L and L-A in the subcutaneous AT (P = 0.057). Adding either linseed or linseed and algae increased α-linolenic acid and eicosapentaenoic acid contents in both AT (P < 0.001); however, docosahexaenoic acid was increased by L-A (P < 0.001). The n-6/n-3 ratio decreased in L and L-A (P < 0.001). Algae had adverse effects on meat quality, with greater lipid oxidation and reduced ratings for odor and flavor. The expression of lipogenic genes was downregulated in the subcutaneous AT (P < 0.05): acetyl-CoA carboxylase 1 (ACACA) in L and L-A and lipoprotein lipase (LPL) and stearoyl-CoA desaturase (SCD) in L-A. Fatty acid desaturase 1 (FADS1), fatty acid desaturase 2 (FADS2) and fatty acid elongase 5 (ELOVL5) were unaffected. In the subcutaneous AT, supplementing either L or L-A increased peroxisome proliferator-activated receptor gamma (PPARG) and CAAT-enhancer binding protein alpha (CEBPA) (P < 0.05), although it had no effect on sterol regulatory element-binding factor 1 (SREBF1). In the intramuscular AT, expression of ACACA, SCD, FADS1 and FADS2 decreased in L and L-A (P < 0.001) and LPL in L (P < 0.01), but PPARG, CEBPA and SREBF1 were unaffected.