Laboratory Report

DISCUSSION Microorganism are organism that are too small and cannot be seen with naked eyes. The phrase of ubiquity of microorganism refers to the concept that microorganism are everywhere in our daily life surrounding. In our everyday common life ,microbes are virtually ubiquitous. They are in the air we breath,the foods we eat and as well as the skin of our fingers. Aseptic transfer is the transference of bacteria or other microbial cultures fromone container to another while maintaining purity of the culture. Pure cultures–consistof only one type of bacteria ideally the descendants from a single bacterial cell.

Because microbes are present everywhere – in the air, the work area, clothes, bodies,etc. , – it is important to follow the rules for aseptic transfer at all times. This is the onlyway of controlling Contamination–Maintaining purity of culture is essential in microbiology if the biologist is to beable to identify bacteria, test for antibiotic sensitivity, or maintain stock cultures. Oftenin nature a pure culture is impossible to come by because species live together. Thescientist is left working with mixed cultures.

Pure cultures can be derived from mixedcultures through isolation of cultures and this also requires that sterile (aseptic)techniques to be used. Normally transference is done from colonies. A colony consists of usuallyseveral million cells that are assumed to be the descendants from one cell. Inoculations from one media to another, therefore, is usually done by removal of a fewmillion cells from one colony into a new environment. This must be done with theintegrity of all colonies remaining intact. Through the use of sterile techniques, this canbe accomplished successfully.

There are a number of tools that are used for inoculation procedures. Inoculating loops are used when transferring members of a broth culture to another broth, platedmedia or an agar slant. Inoculating needles-are used when inoculating a broth culturefrom a colony on plated media or when making a stab in an agar deep or agar slantfrom broth or solid media. Forceps -are used to place sterile disks containing sometesting agent in a broth culture or on a solid media culture. Pipets-are used when transferring liquids into other liquids or onto solid media.

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Flaming-is used to incinerate any microbes left on loops and needles. Alcohol flaming-is used to sterilize forceps. When flaming inoculating loops and needles, careshould be taken avoid burning the plastic handle at the end of each. The metal of theloop or needle should glow red hot and then be allowed to cool before dipping it into any cultures – if the metal is too hot it will kill the organisms that are to be used for inoculation. Alcohol flaming for the forceps is done by dipping the forceps into a smallamount of alcohol and then burning the alcohol off.

The forceps should be dipped andburned three times. Care should be taken to avoid alcohol running up toward the hand. The flame will follow the alcohol and burns will result. Pipets normally used in lab are prepackaged, sterile, disposable pipets. Sometimes glass pipets are used and these are stored in cans. The glass pipets arediscarded into a pipet jar filled with disinfectant. Disposable pipets are deposited inbiohazard bags. It is important that pipettors are always used and pipetting by mouth isprohibited.

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