Bovine tuberculosis (BTB), caused by
Mycobacterium bovis
, is a disease that was introduced relatively recently into the Kruger National Park (KNP) lion population. Feline immunodeficiency virus (FIV
ple
) is thought to have been endemic in lions for a much longer time. In humans, co-infection between
Mycobacterium tuberculosis
and human immunodeficiency virus increases disease burden. If BTB were to reach high levels of prevalence in lions, and if similar worsening effects would exist between FIV
ple
and BTB as for their human equivalents, this could pose a lion conservation problem. We collected data on lions in KNP from 1993 to 2008 for spatio-temporal analysis of both FIV
ple
and BTB, and to assess whether a similar relationship between the two diseases exists in lions. We found that BTB prevalence in the south was higher than in the north (72 versus 19% over the total study period) and increased over time in the northern part of the KNP (0–41%). No significant spatio-temporal differences were seen for FIV
ple
in the study period, in agreement with the presumed endemic state of the infection. Both infections affected haematology and blood chemistry values, FIV
ple
in a more pronounced way than BTB. The effect of co-infection on these values, however, was always less than additive. Though a large proportion (31%) of the lions was co-infected with FIV
ple
and
M. bovis
, there was no evidence for a synergistic relation as in their human counterparts. Whether this results from different immunopathogeneses remains to be determined.