Anaerobic Bacteria Methods for excluding oxygen 1 Fluid

Anaerobic Bacteria

Methods for excluding oxygen 1. Fluid Thioglycollate Media (FTM) • supports aerobic & anaerobic bacteria. • Sodium thioglycollate: removes O 2 from the medium • Resazurin as an oxidation reduction indicator. dye that turns pink in presence of O 2 Mohammed laqqan

Mohammed laqqan

2 - Anaerobic Jar Inoculate FTM tube : B. subtilis or P. aeruginosa (strict aerobes) S. pyogenes (microaerophile) E. coli (facultative anaerobe) C. sporogenes (strict anaerobe) • Place in anaerobic jar. • Incubate overnight at 37 C. Mohammed laqqan

Anaerobic Jar Mohammed laqqan

Anaerobic Jar • For this exercise, we use the Gas. Pak anaerobic system. This system consists of a polycarbonate jar, a lid with a gasket to prevent air flow, a strip infused with methylene blue, and a pouch containing sodium borohydride, sodium bicarbonate, citric acid, and a palladium catalyst. • When water is added to the pouch, the sodium borohydride, sodium bicarbonate, and citric acid react to form hydrogen and carbon dioxide. The palladium catalyzes a reaction between the hydrogen and the oxygen within the jar; this reaction creates water, which forms as condensation on the inside of the jar. • Methylene blue is blue in the presence of oxygen but is colorless in an anaerobic environment. When the oxygen is converted to water and condensation forms on the side of the jar, the indicator strip will turn from blue to white. Mohammed laqqan

Candle jars • The candle flame will consume most of the oxygen in the jar. • Produce an elevated level of carbon dioxide. • These conditions are ideal for the growth of microaerophilic organisms. Mohammed laqqan

Growing Microaerophilic Microbes CO 2 Generating Packet Mohammed laqqan

Anaerobic Chambers • The Vinyl Anaerobic Chambers provide a strict anaerobic atmosphere of 0 -5 parts per million (ppm) using a palladium catalyst and hydrogen gas mix of 5%. Mohammed laqqan

The End Mohammed laqqan