Culture Media Sputum examination checks the quality and quantity of mucus. Visual examination of the sputum determines whether the sputum sample is lacy or thin or contains mucopurulent or saliva. They are contaminated with blood.
Culture Media:
Medium: This is a preparation in which bacteria grow. There are nutrients for bacteria.
Culture Medium: A medium in which bacteria have been added. It is essential for the growth and recognition of bacteria. Different bacteria form colonies of various types and colors.
Blood Agar And Chocolate Agar
This is the most commonly used medium for the growth of certain types of bacteria on which H-influenza is grown. In addition to this to grow Staphylococcus, Pneumococcus, and Streptococcus
It is used for Crystal Violet is added to this medium and it becomes a special type of media on which Streptococcus can be grown.
Chocolate Blood Agar
If blood agar is heated, the R.B.C. inside it breaks down and its color turns brown, in this condition, it is called chocolate agar. Some factors are necessary for its development.
- Preparation Nitrate agar 500mg
- Sterile Blood
- Nitrate agar 2.1g/mm
The prepared nitrate agar is placed in a water hand at 50°C. When the temperature reaches 50°C, blood is slowly added and shaken, followed by 15 mm of med in each dish. or pour to cool and leave to set and cover and refrigerate when cool Bring to normal room temperature when using.
This Stain was introduced by a scientist in 1884. Although the exact mechanism of gram stain cannot be known, it has been found that gram-positive bacteria take color during staining and do not become colorless when acetone is added. They become colorless on pouring and take on the color of the stain that is added later. This difference is between both types. It is caused by bacteria present in cell membranes.
Equipment needed:
- Crystal violet
- Iodine solution
- Lugal iodine solution
- Amarshan oil
- Microscope
- Gram stain
- Acetone
Procedure
Mix the sample well take a little of the material place it in the middle of the slide and make a thick blood film. Then pass the slide through the flame three times to fix the slide. Now place this slide on Costain and cover it with a crystal violet stain. Leave this stain on the slide for 30 to 60 seconds. After that, wash the slide lightly.
Now cover the slide with Lugal iodine Solution. Now wash it for 30 seconds. Now put this slide in Acetone or Acid Alcohol for 30 seconds. Clean with water. Finally, cover the slide with Neutral Red and let the stain stand for two minutes. After two minutes, wash the slide with water dry it, now examine it under a microscope.
Gram Positive Bacteria.
- Stephlycocci
- Streptococci
Some Important Culture Media and Their Preparation.
Alkaline Peptone Water:
This media is of a transparent type, usually for the cholera bacterium Vibra Cholerae
is used.
Method of preparation:
1. Distilled water 300 mm
2. Petone 5 grams
3. Sodium Chloride 5 grams
Its pH is kept between 8 and 01. If the pH is increased, 10/N sodium hydroxide is added and if the pH is decreased, 10/N hydrochloric acid is added. is poured in such a way that there is enough for 50 bottles, Bottles are autoclaved (the lid is softened) and kept at 121°C for 19 minutes and the lid is put on the bottle and the label is placed on the bottle. And it is kept in a cold place. It does not spoil for 2 years.
Cooked Meat Medium:
It is a broth media, it promotes the rapid growth of bacteria and provides them with adequate food.
- Minced meat (heart herb) 454 grams
- Sodium chloride 10 grams
- Petone 5 grams
- Glucose 2 grams
One gram of granulated saffron is placed in a bottle, and 10 ml of distilled water is added and kept for 5 minutes to complete the process. The media is ready.
It is used for the gram-positive bacteria Clostridium.
Lowenstein Jense Acid Medium:
It is a special type of medium used for Mycobacteria, especially Mycobacterium Tuberculosis.
- Chicken eggs 8 numbers
- Hydrochloric acid 8 mm
- Solid glycerol solution 150 mm
- Melajite Green 206/2 3 mm
- Pislin CL 25000
Mix all these ingredients and add 4 ml per bottle into a screw cap bottle, then autoclave and keep the slop and screw the cap. Mycobacterium Tuberculosis Grows in 21 days and colonies appear.
Macconkeys Agar:
It is a specially selected medium that can differentiate between Lactose Fermenter and Non-Lactose Fermenter types of bacteria. Bacteria can grow well on it.
- Pepton 20 grams
- Lactose 10 grams
- Bile salt 5 grams
- Sodium chloride 5 grams
- Neutral Red 0.075 g
- eggs 12 grams
Dissolve all the above-mentioned ingredients in one liter of distilled water and autoclave it. When it gets cold, put it in a petri dish and store it in the refrigerator. This medium does not spoil for about 4 weeks.
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