We evaluated two ELISA assays for detecting bovine antibodies to Mycobacterium paratuberculosis which were adapted for detecting llama and alpaca (LA) antibodies to M. paratuberculosis. A protoplasmic antigen-based assay (Allied Mptb-ELISA) which provides an index score and a whole cell lysate antigen-based assay (IDEXX Mptb-ELISA) which provides an OD value were used. Among a herd of 100 LAs classified as culture negative (CN) for M. paratuberculosis and with no history of exposure to the organism, 52 false-positives occurred with the Allied Mptb-ELISA (48% specificity) and 8 false-positives occurred with the IDEXX Mptb-ELISA (92% specificity). For CN LA, the range for Allied Mptb-ELISA scores was 0.1 - 7.1, and the range for IDEXX Mptb-ELISA OD values was 0.034 - 0.384. For the Allied Mptb-ELISA, there was no significant difference (P=0.48) in the distribution of values for CN LA and 9 culture positive (CP) LA, and no CP LA had values greater than 4.7. In contrast, preliminary results indicate that increasing values for the IDEXX Mptb-ELISA are more closely associated with CP LA than are CN LA. For both ELISAs, males and adults (>2 y) had significantly higher values than females and juveniles (<1 y), respectively. This suggests that age and sex specific cutoffs may be needed for interpreting serologic results. Further evaluations of the IDEXX Mptb-ELISA are currently being conducted, and will include determination of the relative performance of a protein G and an anti-llama IgG conjugate. Until additional evaluations can be completed with larger sample sizes, bovine M. paratuberculosis ELISAs adapted for use on LA should be interpreted with caution. Mptb-ELISA values classified as positive should be confirmed by culture.