This article is available online at http://www.jlr.org ing in cell membranes. Specifi c functions and variations of the various phospholipids (PL), the most abundant lipids in eukaryotic cell membranes, are, however, still poorly understood ( 1 ). A diversity of PL in a fi nely balanced equilibrium is used by cells to construct stable and functional membranes, and PL composition determines most of the physico-chemical cell membrane properties such as fl uidity, permeability and thermal phase behavior ( 2 ).Knowledge of the function of lipids within the cell has benefi ted from the development of increasingly sensitive and selective analytical techniques, particularly those based on mass spectrometry ( 3, 4 ). Among these techniques, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) ( 5 ) has been very successful in studies of the compositions of lipids and other crucial biological molecules. MALDI-MS has allowed direct analysis of complex and unfractionated samples, such as the study of peptide profi les below the level of a single cell ( 6 ). In lipidomics, MALDI-MS has provided fast and simple acquisition of mass spectra with lipid profi les of cells, tissues and body fl uids ( 7 ).MALDI-MS lipid fi ngerprinting can, for example, help studies aimed at understanding the effect of membrane lipid composition on cell membrane behavior after temperature changes. This knowledge is essential for cryopreservation studies of a variety of cells, including oocytes The double molecular layer of polar lipids is a marvelous architectural feature of exquisite biological engineerThis work was supported by the Brazilian research foundations FAPESP (Grant 2008/10756-7) and CNPq.

The objectives were to evaluate the effects of injectable vitamin E during the last 3 wk prepartum on the incidence of retained fetal membranes (RFM) and reproductive performance. Dairy cows (n=890), 390 Holsteins (132 nulliparous and 258 parous) and 500 crossbred Holstein × Gyr (199 nulliparous and 301 parous), from 3 dairy farms in Brazil were assigned to the study. In all 3 farms, from October to March, prepartum cows grazed tropical grasses and received 2 kg/d of a mixture of finely ground corn, soybean meal, and minerals and vitamins. From April to September prepartum cows received a total mixed ration composed of corn silage, finely ground corn, soybean meal, and minerals and vitamins. During the prepartum period, cows were fed 280 (farm 1), 390 (farm 2), and 480 IU (farm 3) of supplemental vitamin E per day, and throughout postpartum, cows were fed 370 (farm 1), 500 (farm 2), and 600 (farm 3) IU of supplemental vitamin E. Within each farm, cows were randomly assigned to remain as untreated controls or to receive 3 i.m. injections of 1,000 IU each of dl-α-tocopherol administered at 19.2 ± 4.3, 12.9 ± 3.3, and 6.2 ± 2.9 d before calving (VitE). Blood was sampled from 141 cows immediately before enrollment to determine the α-tocopherol and cholesterol statuses. Blood was also sampled and analyzed for concentrations of cortisol and nonesterified fatty acids in the last 3 wk of gestation. The serum concentration of α-tocopherol or α-tocopherol:cholesterol ratio did not differ between treatments and averaged 2.97 ± 0.10 μg/mL and 4.46 ± 0.16 × 10(-3), respectively. In total, 53.2% of the cows had an inadequate concentration of serum α-tocopherol based on the 3.0 μg/mL cut-off for adequacy. The risk of RFM decreased as serum α-tocopherol increased. Milk production did not differ between controls and VitE cows. Treatment with injectable α-tocopherol decreased RFM from 20.1 to 13.5%, decreased incidence of stillbirth from 14.9 to 6.8%, and tended to decrease death by 200 d postpartum. VitE cows tended to have improved pregnancy per insemination at first AI (36.7 vs. 30.1%) because of decreased pregnancy loss from 31 to 62 d of gestation (12.5 vs. 20.5%). Despite a similar insemination rate, VitE cows had 22% greater pregnancy rate than control cows. Cows receiving vitamin E had decreased circulating cortisol and nonesterified fatty acids around calving. In summary, when cows were fed limited amounts of supplemental vitamin E, 28 to 48% of the recommendations, prepartum supplementation with injectable α-tocopherol decreased incidence of RFM and improved reproduction.

The objective of this study was to test the hypothesis that high-producing dairy cows become increasingly resistant to insulin throughout lactation and that, consequently, oocyte quality is compromised. We used Holstein cows at 50 (51.5±3.7; n=30), 100 (102.3±9.4; n=30), and 150 (154.5±18.9; n=30) days in milk (DIM). We measured circulating insulin and glucose and performed a glucose tolerance test (GTT) after 5h of fasting. To evaluate oocyte quality, we performed ovum pickup on the day before the GTT (581 oocytes). We performed statistical analyses using the MIXED procedure of SAS. The model included the fixed effects of DIM, period, time, parity, and an interaction between DIM and time. We observed no difference in the GTT between groups for any variable related to circulating glucose (for example, glucose peak=203.3±7.2, 208.8±6.3, and 194.3±5.9mg/dL). However, various measures of circulating insulin were different in cows at 150 DIM compared with 50 or 100 DIM: higher basal insulin (8.8±0.9, 8.8±0.8, and 11.9±0.8 µIU/mL), peak insulin (61.9±6.2 69.1±5.7, and 89.0±6.1 µIU/mL), delta maximum insulin (51.1±5.5 59.4±5.0, and 73.5±5.4 µIU/mL), and area under the curve 5-60 (1,874.8±171.0 2,189.5±157.8, and 2,610.5±174.0 µIU/mL × min). Nevertheless, we observed no difference among groups in the number of viable oocytes (3.2±0.7, 3.9±0.7, and 3.6±0.7 per cow per ovum pickup) or percentage of viable oocytes (49.3, 52.2, and 51.8%). Increased circulating insulin before and throughout the GTT in cows at 150 DIM indicates that cows develop increasing insulin resistance with increasing DIM; however, increased insulin resistance was not associated with a detectable alteration in the quality of oocytes aspirated from small and medium-sized follicles.

Dietary rumen-protected fat rich in linoleic acid may affect the superovulatory response and embryo yield; however, its effects on in vivo embryo cryotolerance are unknown in zebu cattle. The present study evaluated the production and cryotolerance after freezing or vitrification of embryos from Nelore heifers supplemented with rumen-protected polyunsaturated fatty acids (PUFA). Forty heifers kept in pasture were randomly distributed into two groups according to the type of feed supplement (F, supplement with rumen-protected PUFA, predominantly linoleic; C, control fat-free supplement with additional corn). Supplements were formulated to be isocaloric and isonitrogenous. Each heifer underwent both treatments in a crossover design with 70 days between replicates. After 50 days feeding, heifers were superovulated. Embryos were evaluated morphologically and vitrified or frozen. After thawing or warming, embryo development was evaluated in vitro. There was no difference between the F and C groups (P>0.10) in terms of embryo production. Regardless of the cryopreservation method used, Group C embryos had a greater hatching rate after 72h in vitro culture than Group F embryos (44.3±4.2% (n=148) vs 30.9±4.0% (n=137), respectively; P=0.04). Moreover, vitrified and frozen embryos had similar hatching rates (P>0.10). In conclusion, dietary rumen-protected PUFA rich in linoleic acid did not improve embryo production and compromised the cryotolerance of conventionally frozen or vitrified embryos from Nelore heifers.

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