Todd hewitt broth medium

Manufactured by Thermo Fisher Scientific

Sourced in Australia

Todd-Hewitt broth medium is a nutrient-rich culture medium used for the isolation and cultivation of various bacteria, particularly streptococcus and other fastidious organisms. It provides the necessary nutrients and growth factors required for the optimal growth of these microorganisms.

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Lab products found in correlation

Freestyle cho expression medium, by Thermo Fisher Scientific (1 mentions) Columbia blood agar cba, by Thermo Fisher Scientific (1 mentions)

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Todd-Hewitt broth (61 citations) Todd-Hewitt medium (3 citations)

2 protocols using todd hewitt broth medium

Streptococcus pyogenes Strain Culture

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S. pyogenes strains AP1 (S. pyogenes strain 40/58 serotype M1) and AP18 (strain 8/69, serotype M18), both from the WHO Collaborating Centre for Reference and Research on Streptococci, Prague, Czech Republic), MC25 (lacks M1 protein;(36 (link))), BM27.6 (lacks protein H and M1;(37 (link))) and BMJ71 (lacks protein H and M1, SIC, C5a peptidase;(38 (link))) were grown in Todd-Hewitt broth medium (Oxoid) overnight at 37°C, 5% CO₂ without shaking. Mutant strains were cultured in medium supplemented with 150μg/ml kanamycin (MC25), 1μg/ml erythromycin (BM27.6) or 5 μg/ml tetracycline (BMJ71). Overnight bacteria cultures were diluted to OD₆₀₀=0.1 in fresh medium and grown with same conditions until exponential phase growth was reached at OD₆₀₀=0.3–0.4. After centrifugation and washing with 1× PBS, bacteria were ready to use.
Freestyle CHO-S suspension cells (Life Technologies) were used for expression of fusion proteins. Cells were cultured in FreeStyle™ CHO Expression Medium (Thermo Fisher) supplemented with 8mM L-Glutamine (Hyclone) at 37°C, 8% CO₂, 130 rpm shaking.

Blom A.M., Magda M., Kohl L., Shaughnessy J., Lambris J.D., Ram S, & Ermert D. (2017). Factor H – IgG chimeric proteins as a therapeutic approach against the Gram-positive bacterial pathogen Streptococcus pyogenes. Journal of immunology (Baltimore, Md. : 1950), 199(11), 3828-3839.

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Preparation of S. pyogenes for Skin Challenge

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S. pyogenes pNS1 (emm100) was obtained from the Menzies School of Health Research (Darwin, NT, Australia). The isolate had previously been serially passaged in mice to ensure virulence. To prepare for challenge, the isolate was grown overnight in a liquid Todd-Hewitt broth medium (Oxoid, Australia) supplemented with 1% yeast and 1% neopeptone (Difco, Australia). To determine the number of colony forming units (CFUs), the overnight culture was 10-fold serially diluted, plated in duplicate on Columbia Blood Agar (CBA; Oxoid, UK) and supplemented with 5% defibrinated horse blood (Equicell, Australia). The culture was then adjusted to obtain the optimal inoculum^{30 (link)} for skin challenge.

Mills J.L., Jayashi Flores C.M., Reynolds S., Wun C., Calcutt A., Baker S.B., Murugappan S., Depelsenaire A.C., Dooley J., Fahey P.V., Forster A.H., Pandey M, & Good M.F. (2020). M-protein based vaccine induces immunogenicity and protection from Streptococcus pyogenes when delivered on a high-density microarray patch (HD-MAP). NPJ Vaccines, 5, 74.

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