Synovial fluid (SF) was collected at 2, 12 and 26 h post racing from 5 Thoroughbred horses (6 joints) with degenerative joint disease. The effects of serial arthrocentesis on SF TNF alpha levels were controlled for by testing, in parallel, site- and time-matched samples from clinically normal horses (i.e. without arthritis). A significant induction in TNF alpha bioactivity was detected in SF from arthritic joints (compared to the control joints) over the 26 h following racing. After subtraction of values for the arthrocentesis control SF, TNF alpha and protein levels and WBC and mononuclear cell numbers each peaked at 12 h in the SF from the degenerative joints, although there were no statistically significant correlations between any of these parameters. The presence in the SF of TNF alpha, as well as immunoreactive IL-1 beta and IL-6, was confirmed through use of specific anti-human cytokine IgG antibodies in neutralisation and slot-blot radioimmunoassays. TNF beta was not detected in the SF by slot-blot radioimmunoassay. These results suggest that a significant increase in intra-articular TNF alpha occurs during acute inflammatory arthritis in horses. The lack of correlation between infiltrating inflammatory cells and SF TNF alpha levels further suggests that the source of TNF alpha may be resident cells of the joint, as opposed to infiltrating cells found within the joint fluids. SF from clinically normal and arthritic joints of racing and hospitalised horses were also screened for bioactive TNF alpha.(ABSTRACT TRUNCATED AT 250 WORDS)