J. A. Mendizabal

Effect of whole linseed and rumen-protected conjugated linoleic acid enriched diets on feedlot performance, carcass characteristics, and adipose tissue development in young Holstein bulls

Forty-eight young Holstein bulls (slaughtered at 458.6±9.79 kg body weight) were used to evaluate the effect of whole linseed and conjugated linoleic acid (CLA) supplementation on animal performance, adipose tissue development, and carcass characteristics. The animals were fed with one of four isoenergetic and isoproteic diets: control (0% linseed, 0% CLA), linseed (10% linseed, 0% CLA), CLA (0% linseed, 2% CLA), and linseed plus CLA (10% linseed, 2% CLA). Animal performance and carcass characteristics were unaffected by diet composition. Adding linseed or CLA to the concentrate diet did not result in significant differences in adipocyte size and number or lipogenic enzyme activity. However, while the frequency distribution of subcutaneous adipocyte diameters followed a normal distribution, the frequency distribution of intramuscular adipocyte diameters was not normal in any dietary group (skewness coefficients: 0.8, 1.2, 0.9, 0.8 for control, linseed, CLA, and linseed plus CLA, respectively; P<0.05), indicative of adipocyte proliferation in the intramuscular adipose tissue.

Predicting meat yields and commercial meat cuts from carcasses of young bulls of Spanish breeds by the SEUROP method and an image analysis system.

The SEUROP system is currently in use for carcass classification in Europe. Image analysis and other new technologies are being developed to enhance and supplement this classification system. After slaughtering, 91 carcasses of local Spanish beef breeds were weighed and classified according to the SEUROP system. Two digital photographs (a side and a dorsal view) were taken of the left carcass sides, and a total of 33 morphometric measurements (lengths, perimeters, areas) were made. Commercial butchering of these carcasses took place 24 h postmortem, and the different cuts were grouped according to four commercial meat cut quality categories: extra, first, second, and third. Multiple regression analysis of carcass weight and the SEUROP conformation score (x variables) on meat yield and the four commercial cut quality category yields (y variables) was performed as a measure of the accuracy of the SEUROP system. Stepwise regression analysis of carcass weight and the 33 morphometric image analysis measurements (x variables) and meat yield and yields of the four commercial cut quality categories (y variables) was carried out. Higher accuracy was achieved using image analysis than using only the current SEUROP conformation score. The regression coefficient values were between R(2)=0.66 and R(2)=0.93 (P<0.001) for the SEUROP system and between R(2)=0.81 and R(2)=0.94 (P<0.001) for the image analysis method. These results suggest that the image analysis method should be helpful as a means of supplementing and enhancing the SEUROP system for grading beef carcasses.

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.

The effect of vitamin A supplementation on postnatal adipose tissue development of lambs.

Vitamin A (retinoic acid) is known to be an adipogenic factor influencing both in vitro and in vivo cell development. This study aimed to determine its effect on lamb adipose tissue development during the early phase of postnatal development until 100 d of age. Male lambs (n = 24) of the Rasa Aragonesa breed were used. At birth, lambs were assigned to 1 of 2 experimental groups: 1) the control (C) group, which received feed without vitamin A supplementation, and 2) the vitamin A (V) group, which received a supplement of 500,000 IU/animal twice per week from birth to slaughter. The effect of vitamin A supplementation was studied at 16.8 +/- 0.35 kg of BW (58 +/- 0.7 d of age) and at 27.8 +/- 0.78 kg of BW (101 +/- 6.5 d of age). The variables of lamb growth, carcass, LM area, and lipid content were analyzed. To study adipose tissue development, the amount of adipose tissue accumulated, the size and number of adipocytes, and lipogenic enzyme activities (glycerol 3-phosphate dehydrogenase, fatty acid synthase, and glucose 6-phosphate dehydrogenase) of the omental, perirenal, and s.c. depots were quantified. Results showed that vitamin A supplementation had no influence on growth, carcass variables, LM area, and lipid content during lamb growth but that the number of adipocytes in the perirenal depot was 30% greater in lambs of the V group (P < 0.05) and that these lambs had smaller adipocytes in the omental and perirenal depots (P = 0.06) at 28 kg of BW (101 d of age). These results suggest that the intake of this level of vitamin A during the whole period of growth of the lambs influenced the processes of hyperplasia and hypertrophy in the different adipose depots, depending on their degree of maturity.

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.

Low-fat beef patties with augmented omega-3 fatty acid and CLA levels and influence of grape seed extract.

The effects of raising the omega-3 fatty acid (FA), conjugated linoleic acid (CLA), or omega-3 FA plus CLA levels on beef by means of dietary supplementation and of adding grape seed extract (250 mg/kg meat product) in beef patties stored at 2 ± 1 °C in aerobic packaging under simulated retail display conditions for 6 d was evaluated by measuring the thiobarbituric acid reactive substances (TBARS), pH, and instrumental color measurement values and by means of sensory analysis. The pH, instrumental color measurements, and sensory attribute values for patties made from beef with augmented omega-3 FA and/or CLA contents were similar to the values for the control patties made from beef from animals fed a conventional diet. Adding GSE lowered oxidation levels on day 6 (P < 0.001) and did not affect the instrumental color or sensory analysis results during the display period. This suggests that omega-3 FA and CLA-augmented beef could be used to make low-fat beef patties having characteristics similar to those of conventional beef patties while being more in keeping with currently recommended nutritional guidelines.

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.