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Microbiology ppt[1]

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This document discusses methods for cultivating anaerobic bacteria. It defines anaerobes as microorganisms that do not require oxygen for growth and can be killed by oxygen. It then describes five common methods for cultivating anaerobes: the candle jar method, pre-reduced media, the anaerobic jar, the anaerobic chamber, and anaerobic bags and pouches. Each method aims to remove oxygen and maintain an oxygen-free environment needed for the growth of anaerobic bacteria.

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z ASSIGNMENT ON PHARMACEUTICAL MICROBIOLOGY TOPIC:- CULTIVATION OF ANAEROBES Presented by: Sunam jangmu sherpa Roll no:19GPC059 2nd year (3rd semester)
 INTRODUCTION  ANAEROBIC CULTURE METHODS 1. Candle jar method 2. Pre-reduced media 3. Anaerobic jar 4. Anaerobic chamber 5. Anaerobic bags and pouches CONTENT
 An Anaerobes is any unicellular or multicellular microorganism that does not require oxygen for growth ,infact they die the presence of oxygen.  Anaerobes generally rely on carbohydrates so, as to get their energy.  Why they die in the presence of oxygen?  The oxygen molecules that are present in the environment gives out a toxic superoxide radicals that produce hydrogen peroxide,That is toxic for the bacteria so, anaerobes die in the presence of o2.  Anaerobes doesn’t contain a superoxide dismutase and catalase enzyme so, they cannot grow in the presence of o2 Whereas aerobes contains those enzymes and destroy the toxic radicals and can grow in the presence of o2.
1.CANDLE JAR METHOD:-  It is one of the simplest method for obtaining carbon dioxide environment for the growth of anaerobes .  Organisms like Neisseria gonorrhea, steptococcus,etc. can be easily grown by this method.  Enclose the innoculated plate in an airtight jar with a burning candle, As the candle burns the oxygen content is reduced leaving a carbondioxide.  Candle itself act as a indicator as conc. of CO2 Increses candle gets extinguished.  Disadvantages: Some oxygen is always left behind and provides a conc. Of Co2 (5to 10%)that stimilates the growth of microaerophilic bacteria.
2.Pre- REDUCED MEDIA:-  In this methods liquid culture media is used .  Liquid culture media is boiled for 10min, over boiling water to remove most of the dissolved oxygen.Then to this media reducing agents are added(cysteine0.1%),ascorbic acid (0.1%),sodiumthioglycollate(0.1%) , to lower the oxygen content. Eg.,:Robertson cooked meat medium.  Lastly O2 Free nitrogen is bubbled through the medium so as to maintain the anaerobic condition.Then the medium is transferred to tubes and tightly stoppered and sterlised by autoclaving.  Then allowed for incubation.  During incubation ,the tube are flushed with O2 Free carbondioxide.
 It is the most reliable and widely used anaerobic method.  The anaerobic jar contain gas pack that liberated H2 which in turn converts O2 to H2O in the presence of the catalyst(palladium).  The reaction formula is( 2H2+O2 gives 2H2O).  In the presence of water, chemicals present inside the sachet i.e.sodium bicarbonate and sodium borohydride reacts chemically producing hydrogen and carbondoixide gas.  It contain Indicator strip (a indicator such as methylene blue ).  Anaerobic indicator strip included to monitor the anaerobic condition as methyleneblue remains colouless in anaerobic condition.  Its disadvantage is plates have to be removed from the jar to be examined.
Microbiology ppt[1]
 It is a plastic anaerobic glove box that contain an atmosphere of Hydrogen,carbondioxide and nitrogen.  There is an air-lock with inner and outerdoors, the culture media is placed within the air- lock with the innerdoor.  Air of the chamber is removed by vaccum pump and replaced with nitrogen through outerdoor .  Later when the box environment gets favorable then the culture media is placed in the incubator present within the box.
 Anaerobic bags and pouches can be used when few samples are to be incubated anaerobically .  The bags or pouches can remove oxygen as they have catalyst and calcium carbonate that can produce anaerobic environment i.e., carbondioxide rich environment .  One or two incubated plates are placed in the bag , oxygen removal system is activated and the bag is sealed and incubated.  The disadvantage of bags and pouches is it can only be used when few samples are to be incubated.
Have a nice Time ahead. THANK YOU

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Microbiology ppt[1]

  • 1. z ASSIGNMENT ON PHARMACEUTICAL MICROBIOLOGY TOPIC:- CULTIVATION OF ANAEROBES Presented by: Sunam jangmu sherpa Roll no:19GPC059 2nd year (3rd semester)
  • 2.  INTRODUCTION  ANAEROBIC CULTURE METHODS 1. Candle jar method 2. Pre-reduced media 3. Anaerobic jar 4. Anaerobic chamber 5. Anaerobic bags and pouches CONTENT
  • 3.  An Anaerobes is any unicellular or multicellular microorganism that does not require oxygen for growth ,infact they die the presence of oxygen.  Anaerobes generally rely on carbohydrates so, as to get their energy.  Why they die in the presence of oxygen?  The oxygen molecules that are present in the environment gives out a toxic superoxide radicals that produce hydrogen peroxide,That is toxic for the bacteria so, anaerobes die in the presence of o2.  Anaerobes doesn’t contain a superoxide dismutase and catalase enzyme so, they cannot grow in the presence of o2 Whereas aerobes contains those enzymes and destroy the toxic radicals and can grow in the presence of o2.
  • 4. 1.CANDLE JAR METHOD:-  It is one of the simplest method for obtaining carbon dioxide environment for the growth of anaerobes .  Organisms like Neisseria gonorrhea, steptococcus,etc. can be easily grown by this method.  Enclose the innoculated plate in an airtight jar with a burning candle, As the candle burns the oxygen content is reduced leaving a carbondioxide.  Candle itself act as a indicator as conc. of CO2 Increses candle gets extinguished.  Disadvantages: Some oxygen is always left behind and provides a conc. Of Co2 (5to 10%)that stimilates the growth of microaerophilic bacteria.
  • 5. 2.Pre- REDUCED MEDIA:-  In this methods liquid culture media is used .  Liquid culture media is boiled for 10min, over boiling water to remove most of the dissolved oxygen.Then to this media reducing agents are added(cysteine0.1%),ascorbic acid (0.1%),sodiumthioglycollate(0.1%) , to lower the oxygen content. Eg.,:Robertson cooked meat medium.  Lastly O2 Free nitrogen is bubbled through the medium so as to maintain the anaerobic condition.Then the medium is transferred to tubes and tightly stoppered and sterlised by autoclaving.  Then allowed for incubation.  During incubation ,the tube are flushed with O2 Free carbondioxide.
  • 6.  It is the most reliable and widely used anaerobic method.  The anaerobic jar contain gas pack that liberated H2 which in turn converts O2 to H2O in the presence of the catalyst(palladium).  The reaction formula is( 2H2+O2 gives 2H2O).  In the presence of water, chemicals present inside the sachet i.e.sodium bicarbonate and sodium borohydride reacts chemically producing hydrogen and carbondoixide gas.  It contain Indicator strip (a indicator such as methylene blue ).  Anaerobic indicator strip included to monitor the anaerobic condition as methyleneblue remains colouless in anaerobic condition.  Its disadvantage is plates have to be removed from the jar to be examined.
  • 8.  It is a plastic anaerobic glove box that contain an atmosphere of Hydrogen,carbondioxide and nitrogen.  There is an air-lock with inner and outerdoors, the culture media is placed within the air- lock with the innerdoor.  Air of the chamber is removed by vaccum pump and replaced with nitrogen through outerdoor .  Later when the box environment gets favorable then the culture media is placed in the incubator present within the box.
  • 9.  Anaerobic bags and pouches can be used when few samples are to be incubated anaerobically .  The bags or pouches can remove oxygen as they have catalyst and calcium carbonate that can produce anaerobic environment i.e., carbondioxide rich environment .  One or two incubated plates are placed in the bag , oxygen removal system is activated and the bag is sealed and incubated.  The disadvantage of bags and pouches is it can only be used when few samples are to be incubated.
  • 10. Have a nice Time ahead. THANK YOU