In vitro sensitivity of hemophilus influenzae and streptococcus pyogenes to co-trimoxazole.

The invitro testing of Hemophilus influenzae and Streptococcus pyogenes for co-trimoxazole sensitivity requires certain "defined" media that have to be free of inhibitory substances. The use of Columbia agar base with Fildes extract for H. influenzae or of blood agar for S. pyogenes may produce "false-resistant" strains. The addition of thymidine phosphorylases in the form of gentlylysed horse blood (2 to 10%) does not remove all inhibitors in those tests, especially where "undefined" agar bases are used, and results in scanty growth of H. influenzae; the addition of more than 2% results in dark plates, making reading of sensitivities difficult. Fildes agar for testing H. influenzae may be made with enriched sheep or horse blood if the proper "defined" agar base is used. The use of Wellcotest or DST (Oxoid) agar is recommended with Fildes extract for H. influenzae or with blood for S. pyogenes for in vitro testing for co-trimoxazole sensitivity. The addition of thymidine phosphorylase in the form of 2% lysed horse blood does not interfere with reading. However, it results in scanty growth of H. influenzae. Proper inoculation of plates is important. The growth on the plates should be light, dense, but not confluent. Heavy growth may render some strains "false-resistant" even when defined media are used. Our results indicate that many of the previously reported resistant strains of H. influenzae and S. pyogenes may have been "false-resistant" because of the use of "undefined" media. We believe that, in view of our results, respiratory infections may be treated with co-trimoxazole until bacteriologic studies prove that this treatment is contraindicated, since H. influenzae and S. pyogenes are usually found sensitive in vitro under proper conditions.