This study aims to determine the fatty acid (FA) composition and compare FA-based nutritional indices/ratios of colostrum and milk obtained on the 30th, 60th, and 90th day of lactation from 27 Murrah and 18 “Murrah × Carabao” crossbred buffaloes. The major saturated FA (SFA) in colostrum and milk is palmitic acid (C16:0), comprising 32–33% by weight of total FAs. Other important SFAs – myristic acid (C14:0), stearic acid (C18:0), and lauric acid (C12:0) – were lower (P < 0.05) in colostrum than in milk. Oleic acid (C18:1-n9c), a monounsaturated fatty acid (MUFA), is the second most abundant FA in colostrum (28.5%) and milk (18.0–18.8%). Arachidonic acid (C20:4-n6, AA) is the dominant polyunsaturated fatty acid (PUFA) in colostrum (0.92%) and milk (0.42–0.45%). Conjugated linoleic acid (C18:2-c9t11, CLA) was higher in colostrum (0.64%) than in milk (0.14–0.16%). Colostrum and milk had a very low PUFA/SFA ratio (0.02–0.06: 1). The linoleic acid (C18:2-n6, LA) to α-linolenic acid (C18:3-n3, ALA) ratio was higher in colostrum (3.21: 1) than in milk (0.62–1.55: 1). The omega-6 (LA and AA) to omega-3 [ALA and docosahexaenoic acid (C22:6-n3, DHA)] or n-6/n-3 ratio was more balanced for milk (1.76-2.34: 1) than colostrum (3.37: 1). Colostrum had lower atherogenicity (2.53 vs. 4.50–4.66), lower thrombogenicity (2.68 vs. 4.48–4.59), and higher health-promoting index (0.39 vs. 0.21–0.22) than milk. The hypocholesterolemic/hypercholesterolemic (h/H) ratio was higher for colostrum (0.64: 1) than milk (0.34–0.36: 1). Except for AA, the FA composition of colostrum and milk were not significantly different between Murrah and “Murrah × Carabao” crosses (P > 0.05).
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