Todd-Hewitt Broth (T20-106)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-GAS is grown on agar plates produced from Todd-Hewitt broth supplemented with 0.2% yeast extract (THY). All steps should be performed using sterile technique. GAS will remain viable on plates for only 5–7 days after streaking if stored at room temperature. GAS does not survive at 4°C.Sterile wood sticks or inoculating loops used.Optimal growth of GAS is seen in 5% CO2; however, GAS will also grow in ambient air albeit a little slow. -Final pH: 7.8 ± 0.2 at 25°C.

Protocol tips

-GAS is grown on agar plates produced from Todd-Hewitt broth supplemented with 0.2% yeast extract (THY). All steps should be performed using sterile technique. GAS will remain viable on plates for only 5–7 days after streaking if stored at room temperature. GAS does not survive at 4°C.Sterile wood sticks or inoculating loops used.Optimal growth of GAS is seen in 5% CO2; however, GAS will also grow in ambient air albeit a little slow. -Final pH: 7.8 ± 0.2 at 25°C.

Publication protocol

Addition of 1.4% agar to growth media allows the generation of solid media plates. GAS grows best on complex “rich” medium such as Trypticase Soy Agar (TSA) supplemented with 5% Sheep Blood, where it typically produces large zones of β-hemolysis (the complete disruption of erythrocytes and the release of hemoglobin). Therefore, S. pyogenes is also called the Group A β-hemolytic Streptococcus (GABHS). GAS is also commonly grown on agar plates produced from Todd-Hewitt broth supplemented with 0.2% yeast extract (THY). Plates derived from Brain Heart Infusion (BHI) also support the growth of GAS. (See Appendix 2C for list of all media recipes found in CPMC.)

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