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12i)

Introduction Pure culture technique Streak plate method

ii) Pour plate method iii) Spread Plate method 3Reference

Pure culture techniqueIntroductionMicro-organism isolated from various substrate or sources (soil air water food sewage decomposing matter, body surfaces) by growing then. Under more or less well defined conditions as culture contain millions or billions of individuals a culture that contain only one kind of micro organisms is called pure culture. A culture which contain more than one kinds of micro organism is called mixed culture if it contain only two kinds of micro organism that are deliberately maintained in association with one another is called a two member culture. Pure culture are essential in order to study any of the following colony character tics biochemical properties morphology staining reactions and immunological reaction or the susceptibility to antimicrobial agents of a particular strain of fungus bacteria or actionomycetes. Two kinds of operations ie. Isolation and cultivation .

Isolation- the separation of particular microorganism from the mixed population

that exists in nature.

Cultivation- the growth of microbial population in artificial environments

(culture media) under laboratory condition. Pure culture of micro-organism that form discrete colnies an solid media may be most simply obtained by one of the modification of the plating method. The three most commonly used method for obtaining pure culture. of micro organism are1 Streak plate 2- Pour plate and spread plate.

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The Streak plate technique

This method involve the separation and immobilization of individual organism on nutrients medium, solidified with agar. Viable organism give rise through growth to a colony. In this method a sterilized bent wives is tipped into a suitable dilute suspension of organisms and then used to make a series of parallel non over lapping streaks on the surface of an already solidified agar plate. The medium progressively dilute with each successive streak so that isolated conies develop along the lines of later streaks. The streak plate method offense a most practical method of obtaining discrete conies and pure culture. It was originally developed by two bacteriologist Loeffler and Gaffkey in the laboratory of Robert Koch.

ii)

Pour plate method

In the pour plate method the mixed culture is diluted directly in tubes of liquid agar medium. The medium is maintained in a liquid state at a temperature of 45 degree centigrate to allow through distribution of the inoculum. The inoculated medium is dispensed intopetri dishes allow to solidify and then incubated . a series of agar plate showing decreasing number of colones resulting from the dilution procedure in the pour plate technique. The pour plate technique has certain disadvantages for inance, some of the organism are trapped benath the surface of the medium when it gets and there

fore both surface and sub surface colonies develop. The sub surface colonies can be transfer to fresh media. Another disadvantage is that the organism being isolate must be able to withstand temporary exposure to the 45 degree centigrate temperature of the liquid agar medium. For intense the pour plate method would be unsuitable for isolating psychrophilic bacteria.

3- Spread Plate methodIn the spread plate method the mixed culture is not diluted in the culture medium it is diluted in a serried of tubes containing a sterile liquid usually water or physiological saline. A sample is removed from each tube placed into the surface of an agar plate and spread evenly over the surface by mean of a sterile bent glass rod. On at least one plate of the series of bacteria will be in numbers sufficiently low as to allow the development of well separated colonies. In contrast to the pour culture technique only surface colonies develop more over the organism are not required to withstand the temperature of liquid agar. Unlike the streak plate technique the pour plate and the spread plate technique may be performed in a quantitative manner to determine the number of bacteria.

ReferenceMicrobiology(Fifth edition)Michel J. PELCZAR, J.R. E.C.S. CHAN NOELR. KRIEG General microbiology(Fifth edition)Roger Y. Stainer John L. Ingraham Mark. L. Wheelis Page R. Painter

AbstractMicro organism isolated from various substrate or sources (soil, air water, food sewage, decomposing matter body surface) by growing them under more or less well defined condition as cultures contains millions or billions of individuals. A pure culture that contain only one kind of micro organism is called pure culture.

BySarita Pal M.Sc. 1st year Roll No- 31