B. Soret

Meat traceability using DNA markers: application to the beef industry.

Consumer concerns about beef demands instruments to assure its traceability. A methodology using DNA markers is proposed for beef identification focussing on a Spanish beef certification, Ternera de Navarra (Beef of Navarra). To validate this methodology the number of markers used and the implications of population structure in individual identification were evaluated. In order to get practical implementation, the sampling levels required, depending on the number of markers and amount of possible fraud, is also discussed. Using at least eight very informative markers the origin of retailed meat is always found independent of genetic population structure. The total control of fraud would be very expensive using large-scale application of DNA analyses and a strategy based on anonymous sampling is proposed.

Influence of sex on cellularity and lipogenic enzymes of Spanish lamb breeds (Lacha and Rasa Aragonesa)

The effect of sex on the size and number of adipocytes and on the lipogenic enzyme activity in different fat depots in Lacha (L) and Rasa Aragonesa (RA) lambs was studied. Male and female L lambs were fed on ewe milk and were slaughtered at 25 and 24 days of age corresponding to 11·4 and 10·9 kg live weight (UN), respectively. Male and female RA lambs were weaned at 58 days (16·0 kg LW) and were then given concentrates and barley straw until slaughtered at 89 and 91 days of age corresponding to 24·5 and 23·1 kg LW, respectively. A number of parameters were studied in omental (OM), mesenteric (MES) and kidney knob and channel fat (KKCF) depots including the amount of fat, the number and size of adipocytes and the activity of the following enzymes: glycerol 3-phosphate dehydrogenase (G3PDH), fatty acid synthetase (FAS), glucose 6-phosphate dehydrogenase (G6PDH) and NADPmalate dehydrogenase (MD). In subcutaneous (SC) and intermuscular (IM) depots, all the former parameters except the adipocyte number were studied. Females of both breeds had higher amounts of adipose tissue than males in the internal fat depots (P as well as larger adipocytes, mainly in the KKCF (P and P for L and RA lambs, respectively) and OM (P in the RA lambs) depots. There were no differences between sexes in the number of adipocytes. The activity of the G3PDH enzyme was higher in females than in males in OM and SC depots (P in L lambs, and in KKCF, IM (P OM and MES (P depots in RA lambs. Thus, the sex effect on adiposity in both breeds studied involved a greater fattening of the females which was consistent with a greater hypertrophy and a higher G3PDH activity .

Breed effects on cellularity and lipogenic enzymes in growing Spanish lambs

Abstract Size and number of adipocytes and lipogenic enzyme activity in different adipose depots in Lacha (L) and Rasa Aragonesa (RA) lambs has been compared. 42 L males were assigned into three different groups and were slaughtered at the following live weights (LW): 11.4 kg (L12), 24.6 kg (L24) and 35.3 kg (L36). Forty-five RA males were also assigned into three groups and were slaughtered at similar LW than L lambs: 11.70 kg (RA12), 24.5 kg (RA24) and 35.8 kg (RA36). L12 and RA12 lambs were fed on ewe milk until they were slaughtered. After weaning, L24, L36, RA24 and RA36 lambs were fed on commercial concentrate and barley straw, both ad libitum, until they were slaughtered. The parameters studied in omental (OM), mesenteric (MES) and perirenal (PR) adipose depots were: the weight of adipose tissue, the number and size of adipocytes and the activity of Glycerol 3-phosphate dehydrogenase (G3PDH), Fatty acid synthetase (FAS), NADP–malate dehydrogenase (MD) and Glucose 6-phosphate dehydrogenase (G6PDH); in subcutaneous (SC) and intermuscular (IM) depots the same parameters were determined except the adipocyte number. Internal adipose tissue weight in RA lambs was higher than in L lambs due to a greater number of adipocytes at 12 kg LW and to larger adipocytes size at 24 and 36 kg LW. Besides the greater adiposity in RA lambs was consistent with a higher G3PDH enzyme activity, which is tightly correlated with triacylglycerol esterification. Nevertheless, FAS activity was higher in L lambs. In conclusion, it is noted that the different intensity of hypertrophy–hyperplasia and enzyme activities observed in the lambs was the cause for the different adiposity that has been found.

Changes in adipose tissue accumulation in Rasa Aragonesa breed lambs during growth and fattening

Changes during growth and fattening in the number and size of adipocytes and in the activity of the lipogenic enzymes glycerol 3-phosphate dehydrogenase (G3PDH), fatty acid synthetase (FAS) and lipoprotein lipase (LPL) were studied in perirenal (PR) and subcutaneous (SO adipose depots of 28 male lambs of Rasa Aragonesa Spanish breed. Three groups of animals were slaughtered at: 32 (s.d. 6) (no. = 10), 89 (s.d. 8) (no. = 10) and 120 (s.d. 8) (no. = 8) days of age. A significant increase in the quantity of fat was observed as the age of the lambs increased ( P P P P

Effects of Addition of Linseed and Marine Algae to the Diet on Adipose Tissue Development, Fatty Acid Profile, Lipogenic Gene Expression, and Meat Quality in Lambs

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.

Expression of genes involved in adipogenesis and lipid metabolism in subcutaneous adipose tissue and longissimus muscle in low-marbled Pirenaica beef cattle.

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 breeds low tendency to accumulate IMF and the breeds 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.

Single nucleotide polymorphisms in the Melanocortin 1 Receptor gene are linked with lightness of fibre colour in Peruvian Alpaca (Vicugna pacos)

Melanocortin 1 receptor (alpha melanocyte stimulating hormone receptor) (MC1R) is a gene-controlling melanogenesis in mammals. However, it is not well characterized in alpacas and its association with colour is not known. The aim of this study was to look for polymorphisms in the MC1R gene in Peruvian Huacaya alpacas and to analyse the relationship between MC1R single nucleotide polymorphisms (SNPs) and the variations in the instrumental measurement of colour of alpaca fibre. Sixty alpaca fibre samples from black, brown, cream and white animals (15 for each colour) were used to extract DNA from hair bulbs. Colour was measured with a spectrophotometer to obtain quantitative values (CieL*a*b*). Sixteen samples, four of each colour group, were sequenced. Eighteen SNP mutations, 10 not previously described, were found in these 16 sequences. Three of them were chosen (c.82A>G, c.865C>T, c.901C>T) to analyse genotypes by PCR-RFLP in the other 44 fibre samples and to determine the association of mutations with instrumental colour. These three polymorphisms showed association with fibre lightness (P < 0.05), although there was no correlation with colour groups.

Adiposity and adipogenic gene expression in four different muscles in beef cattle

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.

Differentiation in vitro of omental and subcutaneous pre-adipocytes from Spanish Lacha and Rasa Aragonesa sheep

Factors responsible for breed- and depot-specific differences in the development of lipogenic enzymes, and hence lipogenic capacity of adipocytes, in sheep adipose tissue have been investigated using a serum-free cell culture system. Effects of insulin, tri-iodothyronine and exogenous lipid on the development in vitro of the lipogenic enzymes glycerol 3-phosphate dehydrogenase (G3PDH), fatty acid synthetase (FAS), NADP-malate dehydrogenase (ME), glucose 6-phosphate dehydrogenase (G6PDH), and isocitrate dehydrogenase (ICDH) in omental and subcutaneous pre-adipocytes from Lacha and Rasa Aragonesa lambs were investigated. Addition of insulin plus tri-iodothyronine caused pre-adipocyte differentiation, which was enhanced by addition of a lipid supplement. G3PDH activities achieved by differentiation of pre-adipocytes in vitro were similar to those found in vivo; furthermore after differentiation in vitro adipocytes from Rasa Aragonesa lambs had a greater G3PDH activity than adipocytes from Lacha lambs, as found in vivo. In contrast activities of FAS, G6PDH and ME achieved by differentiation in vitro were much greater than those found previously in vivo. While breed- and depot-specific changes in G6PDH observed after differentiation in vitro were similar to those observed in vivo, changes in FAS induced in vitro differed from those found during development in vivo. The study shows that pre-adipocytes from Rasa Aragonesa and Lacha lambs have intrinsic depot- and breed-specific differences in their ability to differentiate and express lipogenic enzymes. The combination of insulin, tri-iodothyronine and a lipid supplement appears to be sufficient to account for in vivo G3PDH activities but other factors are required to explain activities of FAS, G6PDH and ME found in vivo.