Anaerobic Culture Techniques.ppt

Anaerobic Culture Techniques 3rd Science Practical Seminar 1 Vincent O’Flaherty Anaerobic microorganisms are widespread and very important Do not require oxygen for growth - often extremely toxic Facultative anaerobes - can grow in the presence or absence of oxygen Obtain energy by both respiration and fermentation Oxygen not toxic, some use nitrate (NO3-) or sulphate (SO42-) as a terminal electron acceptor under anaerobic conditions Obligate (strict) anaerobes - oxygen is toxic to these organisms, do not use oxygen as terminal electron acceptor Archaea such as methanogens and Bacteria, e.g Clostridia, Bacteriodes etc. etc. Microaerophilic organisms - require low levels of oxygen for growth, but cannot tolerate the levels present in the atmosphere Aerotolerant Anaerobes: Metabolism is anaerobic but they are unaffected by the presence of oxygen. Oxygen Toxicity Oxygen is used by aerobic and facultatively anaerobic organisms as its strong oxidising ability makes it an excellent electron acceptor During the stepwise reduction of oxygen, which takes place in respiration toxic and highly reactive intermediates are produced reactive oxygen species (ROS). Anaerobic and Aerobic Respiration ROS production during respiration O2 + e- = O2- superoxide anion O2- + e- + 2H+ = H2O2 hydrogen peroxide H2O2 + e- + H+ = H2O + OH. Hydroxyl radical OH. + e- + H+ = H2O water Organisms that use O2 have developed defence mechanisms to protect themselves from these toxic forms of oxygen - enymes Catalase: H2O2 + H2O2 = 2H2O + O2 Peroxidase: H2O2 + NADH + H+ = 2H2O + NAD+ Superoxide dismutase: O2- + O2- + 2H+ = H2O2 + O2 Anaerobic environments Anaerobic environments (low reduction potential) include: Sediments of lakes, rivers and oceans; bogs, marshes, flooded soils, intestinal tract of animals; oral cavity of animals, deep undrground areas, e.g. oil packets and some aquifers Anaerobes also important i