# Improved Neisseria gonorrhoeae culture media without atmospheric CO2

## TL;DR

Researchers developed improved culture media for Neisseria gonorrhoeae that eliminates the need for CO2, making bacterial culture easier and more accessible.

## Contribution

The development of CO2-independent culture media for Neisseria gonorrhoeae using sodium bicarbonate.

## Key findings

- Adding 0.75 g/l sodium bicarbonate to Oxoid GC agar base improved growth without CO2.
- CO2-independent versions of Chocolate and Thayer-Martin agars also supported good N. gonorrhoeae growth.
- The new media performed as well as standard media in CO2 incubators.

## Abstract

Bacterial culture on solid media is the crucial step in diagnosing Neisseria gonorrhoeae infections and is the gold standard for determining their antimicrobial resistance profile. However, culture of Neisseria spp. can be challenging in resource poor areas, relying on specialist incubators or other methods of supplying 5% CO2 for growth of the bacteria. Even when such incubators are available, the CO2 to run them may be scarce; there were CO2 shortages during the COVID-19 pandemic, for example. Although culture jars with gas packs or candles can be used, these are inefficient in terms of use of incubator space and researcher time. To achieve simplicity in culturing of N. gonorrhoeae, the standard Oxoid GC agar base medium, made with the Kellogg’s glucose and iron supplements was improved with the addition of 0.75 g/l sodium bicarbonate (NaHCO3), which is inexpensive and readily available. This improved media in a standard incubator performed as well as standard Oxoid GC agar media with supplements in a 5% CO2 incubator. Chocolate agar and Thayer-Martin agar with sodium bicarbonate were also developed, with all showing good growth of N. gonorrhoeae without the need for atmospheric CO2.

• Neisseria spp. (N. gonorrhoeae, N. meningitidis) require atmospheric CO
2
to grow.

• Sources of CO
2
may be scarce depending on geography and lab supply availability.

• We have developed GC, Chocolate, and Thayer-Martin media that does not need CO
2
.