Fatty Acid Composition of Phospholipids and in the Central and External Positions of Triacylglycerol in Muscle and Subcutaneous Fat of Beef Steers Fed Diets Supplemented with Oil Containing n6 and n3 Fatty Acids While Undergoing One of Three 48 h Feed Withdrawal Treatments.

This study was designed to determine the effects of dietary oil and feed withdrawal treatments on fatty acid composition of phospholipids of triacylglycerol in pars costalis diaphragmatis muscle and subcutaneous fat from the brisket. A 2 × 3 factorial experiment was conducted with crossbred steers with an initial body weight of 280.5 ± 5.8 kg. Steers were fed either a control or an oil containing diet where 5% of the control diet was replaced with an equal mixture sunflower and flax oil while undergoing one of three feed withdrawal treatments: no withdrawal, a single 48 h withdrawal before initiation of fattening at one year of age, or 48 h withdrawal at 8 wk intervals from weaning to initiation of fattening. At time of processing samples of muscle and fat were obtained and analyzed to determine fatty acid composition. Disproportionate distribution of the fatty acids was observed by diet, feed withdrawal regimen and whether the sample was from muscle or fat. Differences are discussed in detail, and our data suggests a special function for the fatty acids that accumulate in specific positions of the triacylglycerol due to treatment.

Feeding flaxseed in grass hay and barley silage diets to beef cows increases alpha-linolenic acid and its biohydrogenation intermediates in subcutaneous fat.

The objective of the study was to determine temporal fat deposition and fatty acid profiles in beef cows fed hay- or barley silage-based diets, with or without flaxseed. Crossbred cull beef cows (n = 64, >30 mo of age, 620 ± 5 kg) were removed from grassland pastures, randomly assigned to 16 pens, and given ad libitum access to 50:50 (wt/wt, DM basis) forage:concentrate diets containing 0 or 15% ground flaxseed (DM basis, 5.2% added fat). Diets consisted of hay control (HC), hay+flaxseed (HF), barley silage control (SC), and silage+flaxseed (SF). Backfat biopsies were obtained from each cow at 0, 6, and 12 wk, and at slaughter (~20 wk) to assess fatty acid composition. With the exception of feed efficiency, flaxseed × forage interactions were not significant for backfat accumulation or performance parameters. Flaxseed improved (P < 0.01) feed conversion when supplemented to hay-based diet and increased ADG (P = 0.03), resulting in a heavier (P = 0.02) BW. Compared with hay, barley silage increased (P < 0.01) DMI, ADG, and feed efficiency. Subcutaneous fat contained 0.68% n-3 fatty acids at wk 0, and reached 0.68, 0.81, and 0.94% in HF cows after 6, 12, and 20 wk, respectively (Y(n-3) = 0.0133X + 0.6491, r = 0.87). It was 0.67% at wk 0, and reached 0.65, 0.77, and 0.90% in SF cows after 6, 12, and 20 wk, respectively (Y(n-3) = 0.0121X + 0.6349, r = 0.75). In contrast, weight percentage of n-3 fatty acids decreased in HC cows from 0.63, 0.50, and 0.47, to 0.43%, and in SC cows from 0.63, 0.40, and 0.36, to 0.33% over the 20 wk. A forage × flaxseed interaction (P < 0.05) occurred for many of the α-linolenic acid (ALA) biohydrogenation intermediates, including vaccenic acid (C18:1 trans-11) and CLA (combined C18:2 trans-7,cis-9 and cis-9,trans-11) in plasma, and in subcutaneous fat this also included non-CLA dienes. Concentrations of most α-linolenic acid biohydrogenation intermediates were greater when feeding flaxseed with hay. In conclusion, forage source altered plasma concentrations and rate of accumulation of ALA biohydrogenation products in subcutaneous fat from beef cows fed flaxseed. Factors responsible for this response are yet to be defined, but may include forage-mediated changes in ruminal biohydrogenation of ALA, as well as alterations in fatty acid metabolism and deposition.

Obesity, metabolic syndrome, and adipocytes.

Obesity and metabolic syndromes are examples whereby excess energy consumption and energy flux disruptions are causative agents of increased fatness. Because other, as yet elucidated, cellular factors may be involved and because potential treatments of these metabolic problems involve systemic agents that are not adipose depot-specific in their actions, should we be thinking of adipose depot-specific (cellular) treatments for these problems? For sure, whether treating obesity or metabolic syndrome, the characteristics of all adipose depot-specific adipocytes and stromal vascular cells should be considered. The focus of this paper is to begin to align metabolic dysfunctions with specific characteristics of adipocytes.

Inclusion of flaxseed in hay- and barley silage diets increases alpha-linolenic acid in cow plasma independent of forage type.

Feeding flaxseed to cattle may be a means of increasing omega-3 fatty acid levels in ruminant products, but possible interactions with conserved forages have not been investigated. Twelve Holstein cows were used in a replicated 4 × 4 Latin Square experiment. Cows were fed one of four 50:50 forage:concentrate diets (DM basis): hay (hay control, HC), hay plus 15% ground flaxseed (hay-flaxseed, HF), barley silage (silage control, SC), and barley silage plus 15% ground flaxseed (silage-flaxseed, SF). Plasma concentrations of alpha-linolenic acid (ALA) did not differ between SC and HC diets. Flaxseed increased ALA (P < 0.05), but levels were not influenced by forage type. Flaxseed slightly increased 18:2n-6 (P < 0.05) and some n-6 and n-3 elongation and desaturation products, particularly arachidonic acid (ARA) and eicosapentaenoic acid (EPA). Flaxseed also increased C18:0 (P < 0.05) with this increase being greater (P < 0.01) for cows fed SF than HF. Feeding flaxseed also increased plasma C18:1-trans isomers (P < 0.01), predominantly vaccenic acid (VAA, 18:1-t11), with this increase being greater (P < 0.05) in cows fed HF than SF. Although conjugated linoleic acid (CLA) was increased (P < 0.001) with flaxseed it was not influenced by forage type (P = 0.06). Overall, feeding flaxseed increased plasma ALA, EPA, ARA and CLA independently of forage type. Feeding flaxseed with silage, however, resulted in more 18:0, while feeding flaxseed with hay resulted in greater accumulations of plasma 18:1-trans isomers mainly in the form of VAA.

Examination of adipose depot-specific PPAR moieties.

Molecular mechanisms of peroxisome proliferator activated receptors (PPARs) are being defined rapidly, as illustrated by the volume of papers published. Much of the research is directed towards a clinical end-point/application; however, the non-homogeneous nature of adipose depots in laboratory animals is spurring similar research in domestic meat animals (such as beef cattle). Moreover, the size of adipose depots in meat animals remains an attractive feature for using them to obtain cells for PPAR research. Examination of meat-animal depot-specific PPAR moieties may provide novel information about adipocyte regulation that might be extrapolated to all animals.

Perspectives on the formation of an interdisciplinary research team.

As research funding becomes more competitive, it will be imperative for researchers to break the mentality of a single laboratory/single research focus and develop an interdisciplinary research team aimed at addressing real world challenges. Members of this team may be at the same institution, may be found regionally, or may be international. However, all must share the same passion for a topic that is bigger than any individual's research focus. Moreover, special consideration should be given to the professional development issues of junior faculty participating in interdisciplinary research teams. While participation may be "humbling" at times, the sheer volume of research progress that may be achieved through interdisciplinary collaboration, even in light of a short supply of grant dollars, is remarkable.

Effect of a seaweed extract on fatty acid accumulation and glycerol-3-phosphate dehydrogenase activity in 3T3-L1 adipocytes.

This study was to determine the effect of a seaweed Ascophyllum nodosum extract (SE) containing 220 mg g(-1) phlorotannins on differentiation and fatty acid accumulation in differentiating 3T3-L1 adipocytes. 3T3-L1 cells (2 x 10(4) mL(-1)) were seeded to 24-well plates and proliferated to reach confluence and then were treated with media containing 0, 12.5, 25, 50, 75 and 100 mug mL(-1) SE for 8 days. Dexamethasone, methyl-isobutylxanthine and insulin (DMI) were added to the media in the first 2 days to induce cell differentiation. On day 8 the adipocytes were harvested for measuring cellular fatty acid concentration and the activity of glycerol-3-phosphate dehydrogenase (GPDH). It was found that treatment with SE increased (P < 0.01, n = 6) cellular myristoleic acid (C14:1), palmitoleic acid (C16:1) and oleic acid (C18:1) and total monounsaturated fatty acids (MUFA) without significantly affecting the cell number and saturated fatty acid (SFA). Ratios of MUFA/SFA, C14:1/C14:0, C16:1/C16:0 and C18:1/C18:0 in cellular lipids increased (P < 0.05, n = 6) with the SE treatment in a dose dependent manner (P < 0.001). Treatment with 75 microg mL(-1) SE depressed (P < 0.05) cellular GPDH activity. The results indicate that the biological factors in the SE may be involved in differentiation and MUFA accumulation in adipocytes.

Nonionophore antibiotics do not affect the trans-18:1 and conjugated linoleic acid composition in beef adipose tissue.

The common practice in North American feedlot industries is to add antibiotics to the diet to prevent disease and improve both BW gain and feed efficiency. In this study, 240 crossbred steer calves were backgrounded on a 54% silage diet for 80 d and fed a finishing diet consisting of 81% barley grain, 10% barley silage, and 7.5% supplement (DM basis) with and without in-feed antibiotics for approximately 120 d. Calves were assigned to 1 of 5 treatments: a control with no antibiotics, 11 mg/kg of chlortetracycline, 44 mg/kg of chlortetracycline, 44 mg/kg of chlortetracycline plus 44 mg/kg of sulfamethazine, and 11 mg/ kg of tylosin phosphate. A combination of GLC and silver-ion HPLC methods was used to analyze the fatty acid composition of brisket adipose tissue, with emphasis on trans-18:1 and CLA isomers. The inclusion of nonionophore antibiotics in the diet had little effect on the fatty acid composition, except that feeding either 44 mg/kg of chlortetracycline or 11 mg/kg of tylosin caused small increases in 9c-14:1 and 16:0 relative to the control (0.26 and 0.9 g/100 g of total fatty acids, respectively). Likewise, profiles of trans-18:1 and CLA isomers were unchanged by antibiotics, but across treatments the predominant trans-18:1 isomer was 10t-18:1 (where t = trans; 3.22%) at 3 times the concentration of the second most abundant isomer (11t-18:1; vaccenic acid, 1.05%). Rumenic acid (9c,11t-18:2, where c = cis) was the major CLA isomer at 61% of total CLA, followed by 7t,9c-18:2 at 9%. Because no other effects on fatty acid composition were evident, data for trans-18:1 and CLA were pooled across treatments to investigate possible relationships among rumen PUFA metabolites. The total trans-18:1 content in brisket adipose tissue was positively correlated with 10t-18:1, but not with 11t-18:1, whereas the total CLA was positively correlated with 9c,11t-18:2, but not with 7t,9c-18:2. The 7t,9c-18:2 was, however, positively correlated with 10t-18:1 and 6t/7t/8t-18:1 but was negatively correlated with rumenic acid. These metabolic interrelationships suggest the presence of bacterial populations with distinct pathways for PUFA biohydrogenation in which either 10t-18:1 or 11t-18:1 predominate. Overall, the nonionophore antibiotics tested did not appreciably change adipose tissue composition and consequently could not be used to improve the trans-18:1 or CLA profile (i.e., increase vaccenic and rumenic acids at the expense of 10t-18:1).

Effects of dietary sunflower seeds and tylosin phosphate on production variables, carcass characteristics, fatty acid composition, and liver abscess incidence in crossbred steers.

A 2 x 2 factorial experiment with 48 crossbred steers (with Hereford, Angus, and Charolais genetics, and an initial BW of 373 +/- 8.4 kg) was conducted to evaluate the effects of dietary sunflower seeds (SS) and tylosin phosphate (TP) on production factors, carcass characteristics, liver abscess incidence, and fatty acid composition of the muscle (pars costalis diaphragmatis; PCD) and subcutaneous fat. Individually penned steers were fed either a control diet of 84.5% rolled barley, 14% barley silage, and 1.5% mineral and vitamin mix on a DM basis, or an SS diet, in which SS replaced 15% of the diet. Half the animals fed each diet received TP at 11 mg/kg of DM as a top dressing. Interactions were significant for all production factors. A reduction (P = 0.008) in DMI was observed from 10.1 +/- 0.4 kg/d, in steers fed the control diet, to 8.9 +/- 0.3 and 8.6 +/- 0.3 kg/d, in steers fed the SS and SS + TP diets, respectively. Greater (P = 0.014) ADG was observed for steers fed the control diet than for those fed the SS or SS + TP diet (1.4 vs. 1.1 and 1.2, SE = 0.1 kg/d, respectively); however, G:F ratios were greater (P = 0.011) in steers fed the control diets than in those fed the SS diets. Steers fed the control and SS diets had the heaviest and lightest HCW (347 +/- 6.9 vs. 325 +/- 8.4 kg; P = 0.025), respectively. Lean meat yield (%) of steers fed SS was greater (P = 0.117) than in steers fed the control diets, whereas total lean yield [(HCW x lean meat yield)/100] was similar (P = 0.755). Provision of the SS or SS + TP diet eliminated (P = 0.08 for interaction) liver abscesses compared with the 36 and 9% incidence in steers fed the control or control + TP diet, respectively. Fatty acid weight percentages (wt%) followed similar patterns in PCD and subcutaneous fat. Feeding the SS diets led to greater (P = 0.001) wt% of 18:0 and 18:2n-6, but reduced the wt% of 16:0, 9-cis (c)-18:1, and 18:3n-3 in PCD compared with that in steers fed the control diets, but the wt% of 9c,11-trans (t), and 10t,12c CLA were increased (P = 0.001) by 36 and 400% in PCD. Dietary SS increased (P < 0.001) the wt% of trans-18:1 isomers. The 10t-18:1 and 11t-18:1 isomers were the greatest, but dietary TP elevated (P = 0.004) only 10t-18:1, and total trans-18:1 (excluding 11t-18:1) was 0.47 +/- 0.06 g/100 g of PCD. Dietary SS for finishing steers reduced the incidence of liver abscesses without affecting total lean yield of the carcass, with modest increases in trans fatty acids and in potentially beneficial fatty acids (11t-18:1 and CLA).

Effect of conjugated linoleic acid type, treatment period, and dosage on differentiation of 3T3 cells.

This study was conducted to determine effect of CLA and linoleic acid (LA) on cell differentiation, cellular glycerol-3-phosphate dehydrogenase (GPDH) activity, and FA accumulation in differentiating 3T3-L1 cells (3 isomers x 3 treatment periods x 4 doses). The cells were cultured in 24-well plates for proliferation until confluence. Then they were treated with media containing 0, 10, 35, or 70 mg/L (0, 35, 125, or 250 mmol/L, respectively) of LA, cis9,trans11- or trans10,cis12-CLA during early (day 0-2), intermediate and late (day 3-8), or overall (day 0-8) differentiation periods. Dexamethasone, methyl-isobutylxanthine, and insulin were supplemented to the media only for the early period to induce the differentiation. On day 8 of postconfluence the cells were harvested for Oil Red O staining, analysis of GPDH activity, and determination of the FA Concentration. Cellular LA or CLA was found to accumulate in a dose-response manner, mainly during the intermediate/late period. Treatment with trans10,cis12-CLA lowered (P < 0.05) GPDH activity and the concentration of FA including palmitic acid (16:0) and palmitoleic acid (16:1), especially during the intermediate/late and overall periods, or whenever a high dose of 70 mg/L was applied. This also resulted in a higher (P < 0.05) ratio of saturated FA to monounsaturated FA. Treatment with LA or cis9,trans11-CLA lowered cellular FA only when they applied during the early period at a dose of 70 mg/L. The results demonstrated that the inhibitory effects of CLA on differentiation, GPDH activity, and FA accumulation of 3T3-L1 cells are dependent on the isomer type, treatment period, and dose.

Effects of flaxseed on protein requirements and N excretion of dairy cows fed diets with two protein concentrations.

Thirty-eight midlactating Holstein cows averaging 597 kg of body weight (SD = 59) were used to determine the effects of dietary flaxseed on protein requirement and N excretion in urine and feces. Milk yield and composition, intake, and digestibility were also determined. Cows were allotted from wk 20 to 30 of lactation to 1 of 4 TMR containing 1) no flaxseed (control) and 16% protein (MPC), 2) whole flaxseed and 16% protein (MPF), 3) no flaxseed (control) and 18% protein (HPC), and 4) whole flaxseed and 18% protein (HPF). Cows fed high protein diets had greater feed intake than those fed medium protein diets (20.2 vs. 18.4 kg/d), and cows fed no flaxseed had greater dry matter intake than those fed flaxseed (20.1 vs. 18.5 kg/d). Milk yield was lower for cows fed MPF (20.3 kg/d) than for those fed HPC (24.4 kg/d), HPF (24.9 kg/d), or MPC (24.0 kg/d). Milk protein and lactose concentrations were similar for cows fed MPC and HPC, but flaxseed decreased milk protein concentration in cows fed MPF or HPF compared with cows fed the control diets. Milk fat concentration was similar in cows fed diets with or without flaxseed, but it was decreased by higher protein concentration. Digestibility was generally reduced when diets contained flaxseed and lower protein concentration. Dietary protein had no effect while dietary flaxseed increased fecal N excretion. Retention of N was lower in cows fed flaxseed compared with cows fed the control diets. Feeding flaxseed decreased milk concentrations of short- and medium-chain fatty acids and increased those of long-chain fatty acids. Flaxseed had no effect on the dietary requirement of N by midlactating dairy cows.

Value of sunflower seed in finishing diets of feedlot cattle.

The value of sunflower seed (SS) in finishing diets was assessed in two feeding trials. In Exp. 1, 60 yearling steers (479 +/- 45 kg) were fed five diets (n = 12). A basal diet (DM basis) of 84.5% steam-rolled barley, 9% barley silage, and 6.5% supplement was fed as is (control), with all the silage replaced (DM basis) with rolled SS, or with grain:silage mix replaced with 9% whole SS, 14% whole SS, or 14% rolled SS. Liver, diaphragm, and brisket samples were obtained from each carcass. In Exp. 2, 120 yearling steers (354 +/- 25 kg) were fed corn- or barley-based diets containing no SS, high-linoleic acid SS, or high-oleic acid SS (a 2 x 3 factorial arrangement, n = 20). Whole SS was included at 10.8% in the corn-based and 14% in the barley-based diets (DM basis). In Exp. 1, feeding whole SS linearly increased DMI (P = 0.02), ADG (P = 0.01), and G:F (P = 0.01). Regression of ME against level of whole SS indicated that SS contained 4.4 to 5.9 Mcal ME/kg. Substituting whole for rolled SS did not significantly alter DMI, ADG, or G:F (8.55 vs. 8.30 kg/d; 1.36 vs. 1.31 kg; and 0.157 vs. 0.158, respectively). Replacing the silage with rolled SS had no effect on DMI (P = 0.91) but marginally enhanced ADG (P = 0.10) and improved G:F (P = 0.01). Dressing percent increased linearly (P = 0.08) with level of SS in the diet. Feeding SS decreased (P < 0.05) levels of 16:0 and 18:3 in both diaphragm and subcutaneous fats, and increased (P = 0.05) the prevalence of 18:1, 18:2, cis-9,trans-11-CLA and trans-10,cis-12-CLA in subcutaneous fat. In Exp. 2, barley diets supplemented with high-linoleic SS decreased DMI (P = 0.02) and ADG (P = 0.007) by steers throughout the trial, whereas no decrease was noted with corn (interaction P = 0.06 for DMI and P = 0.01 for ADG). With barley, high-linoleic SS decreased final live weight (554 vs. 592 kg; P = 0.01), carcass weight (329 vs. 346 kg; P = 0.06), and dressing percent (58.5 vs. 59.4%; P = 0.04). Steers fed high-linoleic SS plus barley had less (P < 0.05) backfat than those fed other SS diets. No adverse effects of SS on liver abscess incidence or meat quality were detected. Although they provide protein and fiber useful in formulating finishing diets for cattle, and did improve performance in Exp. 1, no benefit from substituting SS for grain and roughage was detected in Exp. 2. Because of unexplained inconsistencies between the two experiments, additional research is warranted to confirm the feeding value of SS in diets for feedlot cattle.

Effects of dietary sunflower seeds on rumen protozoa and growth of lambs.

Linoleic acid-rich sunflower-seed supplements (SSS) were used in two experiments (experiment 1, high-concentrate diets; experiment 2, high-forage diets) to study effects on rumen protozoa and the growth of lambs. Both experiments consisted of four treatments, two with a low-protein diet (120 g/kg) and two with a high-protein diet (160 g/kg). For both diets, one treatment was without (control) and one with the SSS (140 g/kg dietary DM). The lambs were fed ad libitum for 70 and 140 d in experiments 1 and 2, respectively. Thereafter, the digestibility of organic matter (OM), acid-detergent fibre and neutral-detergent fibre were determined for each diet with four lambs, and then all lambs were slaughtered and rumen fluid samples were collected and analysed. The results showed substantial decreases (P < 0.001) or total elimination of protozoa in the rumen fluid of the SSS-receiving lambs. In the first experiment the SSS also decreased (P < 0.05) feed intake, but an increase in average daily gain (P < 0.06) resulted in an improved (P < 0.05) feed:gain ratio. Also, the SSS increased (P < 0.05) the digestibility of fibre. In the second experiment the SSS decreased (P < 0.05) the OM digestibility, feed intake and growth of lambs. It was concluded that the use of sunflower-seed supplementation in high-concentrate diets of ruminants reduces rumen fauna, resulting in savings on dietary protein supplements and an increased digestion of feed.

Effects of biological type and dietary fat treatment on factors associated with tenderness: I. Measurements on beef longissimus muscle.

The objective was to evaluate chemical, mechanical, and sensory attributes associated with tenderness in divergent cattle breeds--Wagyu (W; n = 12), Limousin (L; n = 12) and F1-cross (WxL; n = 12)--fed two dietary treatments (0 or 6% sunflower oil (DM basis)). A randomized complete block repeated measures design in a 3 x 2 factorial arrangement of treatments was used, and effects of breed, diet, block, and associated interactions were tested. Cattle were fed barley-based diets for an average of 259 d. Twenty-four hours postmortem (PM), steaks from the longissimus muscle (LM) were sliced, vacuum-packaged, aged (1, 3, 7, 14, 28, and 56 d PM) at 2 degrees C, and frozen (-40 degrees C) until analyzed. Wagyu steaks had lower (P < 0.05) Warner-Bratzler shear force (WBSF) values than L steaks across all aging times. At 1 d PM, W steaks required slightly more (P > 0.05) force to shear than WxL or L (0.30 and 0.11 kg, respectively); however, by d 14 PM, W steaks required 0.77 kg less (P < 0.05) force to shear than L. Wagyu steaks received higher (P < 0.05) sensory panel sustained tenderness scores at d 14 PM than L. The pH decline was slower (P < 0.05), and temperature decline more (P < 0.05) rapid, in W carcasses than L or WxL carcasses. Breed and diet did not affect (P > 0.10) free calcium levels (FCL) over time (0, 1, 3, 7, and 14 d PM), 0-h calpastatin activity (CA), d-1 percent collagen (OH-PRO), or d-1 collagen cross-linking (HP). Western blot analysis for the presence of the troponin-T (TNT) 30-kDa fragment, conducted only on samples from steers fed the 0% sunflower oil diet, demonstrated more proteolysis by d 3 PM in L than W or WxL. Overall, breed differences in mechanical and sensory measures of tenderness were not explained by FCL, CA, OH-Pro, and HP. Even though the initial appearance of the TNT 30-kDa fragment was greater in L, linear slopes for appearance of TNT degradation product across aging time were greater for W and WxL (P < 0.01 and P = 0.056, respectively) than for L, suggesting that tenderness differences due to breed may have been facilitated by more-rapid proteolytic degradation over time.

Effects of biological type and dietary fat treatment on factors associated with tenderness: II. Measurements on beef semitendinosus muscle.

The objective of this study was to evaluate attributes in semitendinosus muscle (ST) associated with tenderness in divergent breeds--Wagyu (W; n = 12), Limousin (L; n = 12), and Wagyu x Limousin cross cattle (WxL; n = 12)--fed two dietary treatments (0 or 6% sunflower oil, DM basis). A randomized complete block repeated measures design with a 3 x 2 factorial arrangement of treatments was used to measure effects of breed, diet, block, and associated interactions. Cattle were fed barley-based diets for an average of 259 d. Temperature and pH were measured at 0, 1, 3, 6, 12, and 24 h postmortem (PM). Steaks from the ST were removed 24 h postmortem, vacuum-packaged, aged (1, 3, 7, 14, 28, and 56 d postmortem) at 2 degrees C, and frozen (-40 degrees C) until analyzed. Dietary treatment did not (P > 0.10) affect Warner-Bratzler shear force (WBSF), collagen amount (OH-PRO) or cross-linking (HP), temperature, or pH. Steaks from WxL aged 14 d postmortem had lower (P < 0.05) WBSF values than L (W were intermediate). Cooking time was longer (P < 0.01) in W and WxL than in L; however, breed did not affect (P > 0.10) cooking loss. Cooking time was not influenced by diet, but steaks from cattle fed 6% sunflower oil had lower (P < 0.05) cooking losses. Temperature decreased more (P < 0.05) rapidly, and pH more slowly (P < 0.05), in W and WxL than L in the first 24 h postmortem. Limousin steaks were lighter (higher L*) and more yellow (higher b*) in color than steaks from W and WxL (P < 0.05). The control diet (no oil added) resulted in steaks that were lighter (P < 0.05) than the treatment diet (6% added sunflower oil). Neither breed nor diet affected (P > 0.10) OH-PRO or HP concentration. The results of this study indicate that biological type differences may not be as great in the ST as in longissimus muscle; thus, to increase tenderness in ST, emphasis may need to be placed on processing and cooking techniques rather than genetic selection.

Primary adipocyte culture: adipocyte purification methods may lead to a new understanding of adipose tissue growth and development.

In the present manuscript, the methods required to generate purified cultures of mature adipocytes, as well as stromal vascular cells, from the same isolation are detailed. Also, we describe the in vitro conditions for the dedifferentiation of the isolated mature adipocytes. These two types of cells may be used to reevaluate differences between presently available cellular models for lipogenesis/lipolysis and might provide a new cellular physiological system for studies utilizing the proliferative progeny from mature adipocyte dedifferentiation. Alternative possibilities to the dedifferentiation phenomenon are proposed, as this new area of research is novel.

Growth, carcass characteristics, muscle conjugated linoleic acid (CLA) content, and response to intravenous glucose challenge in high percentage Wagyu, Wagyu x Limousin, and Limousin steers fed sunflower oil-containing diet.

The effect of breed and diet on insulin response to glucose challenge and its relation to intramuscular fat deposition was determined in 36 steers with 12 each of greater than 87% Wagyu (referred to as Wagyu), Wagyu x Limousin, and Limousin breeds. Weaned steers were blocked by weight into heavy, medium, and light calves and placed in six pens with two pens per weight type and with two steers of each breed per pen. Three pens with steers from each weightclass were fed backgrounding and finishing diets for 259 d, while the other three pens were fed the same diets where 6% of the barley grain was replaced with sunflower oil. Prior to initiation of the finishing phase of the study the intravenous glucose tolerance test (VGTIT) was conducted in all steers. Once steers were judged as carrying adequate 12th-rib fat, based on weight and days on feed, they were harvested and graded and samples of the longissimus muscle were procured for determination of fat content and fatty acid composition. Dietary oil improved (P = 0.011; 0.06) ADG and feed conversion efficiency of steers during the latter part of backgrounding and only ADG during early part ofthe finishing period. Generally percent kidney, pelvic, and heart fat was the only adiposity assessment increased (P = 0.003) by dietary oil. The IVGTT results indicated that insulin response to intravenous glucose was lower in Limousin steers than in Wagyu steers. Dietary oil decreased (P = 0.052) fasting plasma insulin concentration in Wagyu steers compared with Limousin steers. The correlation coefficients among the IVGTT measures and intramuscular fat content or marbling score were less than 0.4, and only a negative trend existed between fasting insulin and USDA marbling scores. However, the carcasses of the Wagyu steers graded US Choice, and 66% of the Wagyu carcasses graded US Prime, which were substantially better than the quality grades obtained for the carcasses from the other breed types. Dietary oil did not affect muscle fat content but increased (P = 0.01) conjugated linoleic acid (CLA) concentrations by 339%. Results indicated that IVGTT measures were not appropriate indices of marbling potential in cattle and that dietary oil can enhance CLA content of beef.

Characterization of microsomal diacylglycerol acyltransferase activity from bovine adipose and muscle tissue.

The activity of the triacylglycerol bioassembly enzyme, diacylglycerol acyltransferase (DGAT), was characterized in microsomal fractions prepared from bovine subcutaneous (SC) adipose, intramuscular (IM) adipose, and muscle (pars costalis diaphragmatis) tissue. The activity of DGAT was generally higher from SC adipose tissue than from IM adipose or muscle tissue. The characteristics of DGAT activity from the three bovine tissues resembled the activity characteristics observed in previous studies from various other organisms and tissues; the pH optimum was near neutrality, the activity was almost completely inhibited by pre-incubation with N-ethylmaleimide (NEM), and the enzyme accepted a broad range of acyl-CoAs and sn-1,2-diacylglycerols. In some aspects, the SC adipose tissue DGAT activity was different from the DGAT activity from the other two tissues. The SC adipose tissue DGAT activity was not as susceptible to inhibition by NEM as the enzymes from the two other tissue sources, and it exhibited increased specificity for substrates containing oleoyl moieties. The differences in DGAT properties between the three bovine tissues may account to some extent for the differences in the relative fatty acid composition and the positional distribution of fatty acids in triacylglycerol between bovine tissues. The observed differences in enzymatic properties also support recent biochemical and molecular genetic observations that imply the existence of multiple DGAT genes and/or isoforms.

Ten commandments for preventing contamination of primary cell cultures.

Procedures for preventing contamination in primary cell cultures must be carefully defined and strictly followed in order to obtain healthy cells. Protocols have been developed and refined in our laboratory for establishing primary cultures of muscle and fat stem cells without contamination from a variety of animals. Contamination of cell cultures is not only frustrating, but is also very expensive both in time and loss of materials. Through the consistent use of proper aseptic techniques, most instances of contamination may be avoided. We suggest that the basic principles detailed here will find wide applicability in the culturing of primary cells without contamination from many different types of animals and tissues.

Relationship of fatty acid composition to intramuscular fat content in beef from crossbred Wagyu cattle.

The deposition of i.m. fat, or marbling, in cattle is recognized as a desirable carcass trait in North American beef grading schemes. In order to investigate the relationship between degree of marbling and fatty acid composition of whole bovine muscle, we extracted the total lipid from pars costalis diaphragmatis (PCD) (n = 23) and longissimus (n = 36) muscles from Wagyu crossbred cattle that were assigned Canadian Grading Agency marbling scores ranging from 1 to 8 on an inverse 10-point scale (i.e., a score of 1 indicated "very abundant" marbling and a score of 10 would be assigned to a carcass "devoid" of marbling). Fatty acid methyl esters (FAME) of the total lipid and triacylglycerol fractions were resolved and quantified through GLC. Marbling scores were negatively associated with total lipid from both PCD (r = -.57, P < .01) and longissimus (r = -.80, P < .001). Differences between PCD and longissimus were found for almost all FAME studied from both lipid fractions, but no differences (P > .05) were seen when the monounsaturated:saturated fatty acid (MUFA/SFA) ratios were compared. Heifers had higher (P < .05) oleic acid content and lower (P < .05) palmitic acid content in lipid extracted from both muscles, resulting in higher (P < .05) MUFA/SFA ratios than those for steers. The relative amount of myristic acid increased as the lipid content (total lipid and triacylglycerol) increased in either longissimus (r values from .48 to .55; n = 36; P < .01) or PCD muscles (r from .67 to .76; n = 23; P < .001). The relative amount of linoleic acid (cis-9, cis-12 isomer) from total lipid was negatively associated with all chemical measurements of lipid from the longissimus (r from -.52 to -.64; n = 36; P < .001) and PCD muscles (r from -.75 to -.85; n = 23; P < .001). This association was not significant (P > .1) for either muscle when linoleic acid from the triacylglycerol fraction was examined, suggesting the negative association between this fatty acid and lipid content was due to a dilution of membrane phospholipids with increasing triacylglycerol. Indices of fatty acid elongase activity, calculated from FAME data, implicated the balance between this enzyme activity and fatty acid synthase as a source of variation between animals displaying various degrees of marbling and worthy of further investigation to better understand the process of marbling fat deposition in beef cattle.