Haemophilus influenzae Rd [KW20] was obtained from ATCC (51907). Standard growth and culturing techniques were followed as described previously92 (link). Cultures were grown in Brain Heart Infusion broth (BHI) supplemented with 7.5 μM of hemin and 2 μg/ml NAD with or without the addition of peptide for 24 hours. The number of viable cells for every reaction mixture was then determined by serially diluting and spotting 10-μl aliquots in triplicates on BHI agar plates supplemented with 15 μM hemin and 3 μM NAD.
Porphyromonas gingivalis 2561 was obtained from ATCC (33277). Pre–reduced, anaerobically sterilized Brucella Broth and BRU - Brucella Blood Agar – were purchased from Anaerobe systems (CA, USA). They were opened just before use. Static cultures and plates were incubated at 37°C in an incubation chamber from BD GasPak™ EZ Container Systems. Anaerobic conditions were maintained by using BD BBL CO2 gas generators and BD BB GasPak CO2 indicators. Cultures were grown anaerobically in Brucella Broth with or without addition of peptides for 48 hours and viable cells for every reaction mixture were then determined by serially diluting and spotting 10-μl aliquots in triplicates on Brucella Blood Agar.
Porphyromonas gingivalis 2561 was obtained from ATCC (33277). Pre–reduced, anaerobically sterilized Brucella Broth and BRU - Brucella Blood Agar – were purchased from Anaerobe systems (CA, USA). They were opened just before use. Static cultures and plates were incubated at 37°C in an incubation chamber from BD GasPak™ EZ Container Systems. Anaerobic conditions were maintained by using BD BBL CO2 gas generators and BD BB GasPak CO2 indicators. Cultures were grown anaerobically in Brucella Broth with or without addition of peptides for 48 hours and viable cells for every reaction mixture were then determined by serially diluting and spotting 10-μl aliquots in triplicates on Brucella Blood Agar.