We studied the effects of Salmonella typhimurium lipopolysaccharide (LPS) on in vivo cytokine mRNA levels in chickens, and investigated whether these levels could be altered by different nutrients. Two hundred and forty chicks were assigned in a 2 x 4 factorial design of treatments. Factors were intravenous injection with S. typhimurium LPS, or saline (control), and four dietary fat sources: corn oil (CO), linseed oil (LO), menhaden oil and beef tallow (BT). Two hours after injection birds were killed and their spleens removed for RNA extraction. Quantitative real-time RT-PCR assays for mRNA of chicken IFN-gamma, IL-6, IL-8, IL-15, IL-18 and 28S rRNA were used to obtain the in vivo splenic cytokine profiles. Expression levels of IL-6, IL-8, IL-18 and IFN-gamma mRNA increased, but IL-15 mRNA decreased 2h after challenge with LPS compared with saline controls. In saline-injected control chickens, the dietary oil source did not affect the splenic mRNA level of any cytokine. In LPS challenged chickens IFN-gamma mRNA was significantly higher in the chickens fed the fish oil enriched diet compared with the LO, CO and BT enriched diets. The present data imply that avian IL-15 has, at least partially, a different function compared to its mammalian counterpart, and in addition, chicken innate immune responses might be affected differently by n-3 PUFA compared to mammals.