The use of 2-dimensional gas chromatography to investigate the effect of rumen-protected conjugated linoleic acid, breed, and lactation stage on the fatty acid profile of sheep milk

In this study, 2-dimensional gas chromatography (GC x GC) was used to obtain a detailed fatty acid (FA) profile of sheep milk and to evaluate the effects of a rumen-protected conjugated linoleic acid (rpCLA) supply, breed, days in milk (DIM), sampling period, and number of lambs suckling on the FA profile. Twenty-four ewes, from 3 autochthonous breeds of the Veneto Alps (Brogna, Foza, and Lamon), were housed in 6 pens (2 pens/breed), according to DIM (38 +/- 23 d) and body weight (61 +/- 13 kg). The ewes and their offspring of 3 pens (1 pen/breed) were fed ad libitum a total mixed ration (control), and the other animals received the same diet supplemented with 12 g/d per ewe, plus 4 g/d for each lamb older than 30 d, of an rpCLA mixture. The study lasted 63 d. Two composite milk samples for each ewe were prepared during the first and second months of the trial. The pooled milk samples were analyzed in duplicate for FA profile by 2-dimensional gas chromatography, which allowed us to obtain a detailed FA profile of sheep milk, with 170 different FA detected, including many that were present in small concentrations. The milk relative proportions of individual FA, groups of FA, or FA indices were analyzed by PROC MIXED of SAS (SAS Institute Inc., Cary, NC), considering diet, breed, DIM, and sampling period as sources of variation. The random effect of animal was used to test diet, breed, and DIM, whereas the effects of period were tested on the residual. Breed had a small influence on milk FA profile, mainly on branched-and odd-chain FA. Within breed, animal repeatability for the relative proportions of milk FA was notable for almost all monounsaturated FA and for saturated FA with 14 to 19 carbon atoms, except C16:0, and less so for polyunsaturated FA. The inclusion of rpCLA (CLA cis-9, trans-11 and CLA trans-10, cis-12) increased the presence of the same CLA isomers in the milk as well as that of CLA trans-9, trans-11, and decreased the proportions of de novo-synthesized short-chain FA. From a cluster analysis based on the matrix of correlation coefficients among all FA relative proportions, 3 main FA groups were observed: the first included mainly odd-or branched-chain saturated FA, C18:0, C16:0 and CLA trans-10, cis-12; the second included monounsaturated FA or polyunsaturated FA with 16 to 20 carbons, CLA cis-9, trans-11, and CLA trans-9, trans-11; and the third included short-to medium-chain saturated FA, polyunsaturated FA with 2 to 5 double bonds, and 3 CLA isomers not affected by rpCLA addition (CLA trans-11, cis-13, CLA cis-9, cis-11, and CLA cis-10, cis-12)