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Date: 2076/2/26 Experiment No-1: Typical Composition

The document summarizes an experiment on preparing different culture media, evaluating quality, and enumerating microbes from contamination. Potato dextrose agar and nutrient agar were prepared following standard procedures and compositions. Observations showed 3 whitish yellow curled colonies on potato dextrose agar and 1 slightly pink cocci cluster colony on nutrient agar that was determined to be gram-positive bacteria. The experiment demonstrated preparation of common media and basic microbial analysis.

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Umesh Poudel
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59 views3 pages

Date: 2076/2/26 Experiment No-1: Typical Composition

The document summarizes an experiment on preparing different culture media, evaluating quality, and enumerating microbes from contamination. Potato dextrose agar and nutrient agar were prepared following standard procedures and compositions. Observations showed 3 whitish yellow curled colonies on potato dextrose agar and 1 slightly pink cocci cluster colony on nutrient agar that was determined to be gram-positive bacteria. The experiment demonstrated preparation of common media and basic microbial analysis.

Uploaded by

Umesh Poudel
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Date: 2076/2/26

Experiment no- 1
Name of experiment- Preparation of different culture media, quality evaluation and enumeration
of microbes from contamination

Objectives

1. To know about culture media

2. To know about quality evaluation

3. To enumerate the microbes from food

Theory

A growth medium or culture medium is a solid, liquid or semi-solid designed to support the
growth of microorganisms or cells, or small plants like the moss Physcomitrella patens. Different
types of media are used for growing different types of cells. The two major types of growth
media are those used for cell culture, which use specific cell types derived from plants or
animals, and microbiological culture, which are used for growing microorganisms, such as
bacteria or fungi. The most common growth media for microorganisms are nutrient broths and
agar plates; specialized media are sometimes required for microorganism and cell culture
growth. Some organisms, termed fastidious organisms, require specialized environments due to
complex nutritional requirement.

Potato dextrose agar

Potato dextrose agar and potato dextrose broth are common microbiological growth

media made from potato infusion, and dextrose. Potato dextrose agar is the most widely

used medium for growing fungi and bacteria.

Typical composition

1000ml Water

4g (from 200g infused potato) Potatoes (sliced washed unpeeled)

20g Dextrose

15g agar powder

5.6±0.2 Final ph
Nutrient agar

Nutrient agar is a general purpose medium supporting growth of a wide range of non

fastidious organisms. It typically contains (mass/volume)

 0.5% Peptone - this provides organic nitrogen


 0.3% beef extract/yeast extract - the water-soluble content of these contribute vitamins,
 carbohydrates, nitrogen, and salts
 1.5% agar - this gives the mixture solidity
 0.5% Sodium Chloride - this gives the mixture proportions similar to those found in
 the cytoplasm of most organisms
 Distilled water - water serves as a transport medium for the agar's various substances
 pH adjusted to neutral (6.8) at 25 °C (77 °F).

Procedure

Preparation of media plate

1. Potato dextrose agar


 39 g of commercial PDA powder was added to 1000 ml of distilled water.
 Mixture was dissolved by boiling.
 Then, autoclave was done for 15 min at 1210C.
 Finally, the solution was poured in the sterile petri plate and allowed
 to cool. The prepared PDA was incubated at 370C for 48 hours.
2. Nutrient agar
 28 g of nutrient agar was mixed in 1000 ml distilled water.
 Mixture was dissolved by heating.  Media was autoclave at 1210C for 15 min.
 The media was poured in the sterile petri plate and allowed to cool.
 The prepared NA media was incubated at 370C for 48 hours.

3. The incubated plate was observed after 48 hours.

4. Non- contaminated plated were stored at refrigerator and contaminated plateb were placed
for further steps.

5. Simple gram staining was done for bacteria, yeast and mold.
Observation:

PDA colony

No of colony 3

Color of colony Whitish yellow

Morphology of colony Curled, circular

NA colony

No of colony 1

Color of colony Slight pink

Cocci in clusters Cocci in clusters

Gram positive or negative Gram positive


Gram positive

Result and Discussion

Hence, from the above observation 3 colonies were observed having curled and circular

morphology. Also 1 colony was observed in the nutrient agar plate, which seems cocci

in cluster, and is determined as the presence of gram positive bacteria.

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