Thiamine pyrophosphate (cocarboxylase) as a growth factor for Haemophilus somnus

The effect of a commercially available, chemically defined enrichment (Iso-VitaleX; BBL Microbiology Systems, Cockeysville, Md.) on the growth of 10 strains of Haemophilus somnus was studied. A 6- to 10-fold increase in growth, as measured turbidimetrically, was observed when Iso VitaleX was added to a basal medium of brain heart infusion broth to a final concentration of 1% (vol/vol). Thiamine pyrophosphate (cocarboxylase), a constituent component of Iso VitaleX, was found to be the only growth-promoting factor, and it could be used as a substitute for Iso VitaleX. An equimolar concentration (2.2 microM) of thiamine monophosphate promoted growth equal to that of thiamine pyrophosphate. Thiamine was nonstimulatory for all 10 strains tested. When alkaline thermal-treated brain heart infusion broth was used as the basal medium, 7 of the 10 strains had an absolute requirement for thiamine monophosphate or thiamine pyrophosphate. The three remaining strains showed minimal growth when thiamine was added to this basal medium; however, excellent growth was observed when thiamine monophosphate or thiamine pyrophosphate was utilized. Factor X (hemin) was found to further enhance the growth when concentrations of 5 to 10 micrograms/ml were coupled with thiamine pyrophosphate. No increase in growth was observed when factor V (nicotinamide adenine dinucleotide) was coupled with thiamine pyrophosphate. This is the first report of a growth factor requirement for H. somnus.