Alteration of the phospholipid (PL) and triglyceride (TG) content and/or fatty acid pattern was recently documented in the plasma, liver, heart, brain, adipose tissue and muscle of second generation rats depleted in long-chain polyunsaturated w3 fatty acids (w3 rats). The present study extends this knowledge to the submandibular gland. Twelve female control rats (C) were compared to 12 w3 rats of same sex and age. The total PL and TG content of the salivary gland was not significantly different in C and w3. The most abundant fatty acids were C20:4w6, C16:0 and C18:2w6 in the PL and C16:0, C18:1w9 and C18:2w6 in TG. The sole w3 fatty acids found in w3 rats were C22:5w3 and C22:6w3, which in the PL were 3 to 12 times lower than in C rats. This coincided with a C22:5w3/C22:6w3 ratio 3 times higher in w3 than C rats. As judged from the (C16:0 + C16:1w7)/(C18:0 + C18:1w9) ratio in PL and TG, and the C18:0/C20:0, C20:0/C22:0, C22:0/C24:0 and C18:2w6/C20:3w6 ratios in PL, the activity of elongase was comparable in C and w3 rats. The PL C20:4w6/C22:4w6 ratio was twice higher, however, in C than w3 rats. The activity of D9-desaturase, as judged from the PL and TG C16:0/C16:1w7 and C18:0/C18:1w9 ratios was higher in w3 than C rats. No significant difference between C and w3 rats was found in the PL C18:2w6/C18:3w6, C20:2w6/C20:3w6 and C20:3w6/C20:4w6 ratio. Nevertheless the step-wise conversion of C18:2w6 to C20:4w6 appeared facilitated in the w3 rats, as judged from the ratio between these two fatty acids in either PL or TG. Some of the w3 rats were injected intravenously 60–120 min before sacrifice with 1.0 ml of either a medium-chain triglyceride: fish oil emulsion or a control medium-chain triglyceride: olive oil emulsion. The former emulsion increased the amount of C22:5w3 and C22:6w3 in PL. This coincided apparently with a more efficient conversion of C22:5w3 to C22:6w3, at least in young rats. Moreover, the prior injection of the fish oil-containing emulsion prevented the age-related decrease in both the C22:5w3 and C22:6w3 PL content and C16:0/C16:1w7 and C18:0/C18:1w9 ratio otherwise also recorded in PL. In conclusion, these findings, in addition to illustrating the severe depletion in w3 fatty acids of the w3 rats, document an increased activity of D9-desaturase, a more efficient conversion of C18:2w6 to C20:4w6 and C22:4w6 and an apparent impairment of C22:5w3 conversion to C22:6w3 in the same animals.