Microbiology Power Point

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Microbiology Power Point

  1. 1. Microbiology partition Prepared by Mostafa Salah El- Din Biochemistry division Cultures and Sensitivity
  2. 2. 1) The cultures : can be used to detect the microorganisms Which cause an infection in any part of the body such as ; urinary tract , throat, GIT, ear, eye and the respiratory tract; etc . 2) The sensitivity test : can be used to detect the most effective antibiotic against the microorganism that cause a certain infection.
  3. 3. 1) Urine culture. 2) Stool (Fecal) culture. 3) Blood culture. 4) Sputum culture. 5) CSF culture. 6) Pus (Wound) culture. Types of cultures
  4. 4. 7) Semen culture. 8) Prostatic culture. 9) Vaginal culture (swab). 10) Cervical culture. 11) Nasal culture (swab). 12) Eye culture (swab). 13) Ear culture (swab). 14) Throat culture (swab).
  5. 5. . Types of media <ul><li>Nutrient agar medium. </li></ul><ul><li>Mackonkey medium. </li></ul><ul><li>SS agar medium ( Salmonella Shigella </li></ul><ul><li>agar medium). </li></ul><ul><li>Blood agar medium. </li></ul><ul><li>Chocolate agar medium. </li></ul>
  6. 6. <ul><li>Nutrient agar medium : is a suitable medium </li></ul><ul><li>for the growth of both gram positive and gram </li></ul><ul><li>negative bacteria . </li></ul><ul><li>Mackonkey medium : is a suitable medium </li></ul><ul><li>for the growth of gram negative bacteria only, so </li></ul><ul><li>it is used to determine which type of bacteria </li></ul><ul><li>cause the infection. </li></ul><ul><li>The 3 other types of media are suitable for </li></ul><ul><li>the growth of both gram positive and gram </li></ul><ul><li>negative bacteria. </li></ul>
  7. 7. <ul><li>1) Nutrient agar medium : </li></ul><ul><li>a) 7 g of agar dissolved in 250 ml distilled water in a conical </li></ul><ul><li>flask. </li></ul><ul><li>b) Boiling in an oven or on a hot plate. </li></ul><ul><li>c) Then, leave it for cooling in the air. </li></ul><ul><li>d) After the cooling, store in the refrigerator for use. </li></ul><ul><li>2) Blood agar medium : </li></ul><ul><li>Can be prepared by pouring of warmed agar in a Petri dish containing 1- 2 cm blood. </li></ul><ul><li>3) Chocolate agar medium : </li></ul><ul><li>Can be prepared by a pouring of boiled agar in a Petri dish containing 1- 2 cm blood. </li></ul>Preparation of the media
  8. 8. 4) Mackonkey medium : a) 12.8 g of Mackonkey powder dissolved in 250 ml distilled water. b) Boiling in an oven or on a hot plate. c) Leave it for cooling in the air. d) After cooling, store in the refrigerator for use.
  9. 9. <ul><li>Each type of the cultures must be performed on </li></ul><ul><li>a suitable medium for the growth of the micro- </li></ul><ul><li>organisms that cause the infection, as follow ; </li></ul><ul><li>A) Urine sample can be cultured on two media: </li></ul><ul><li>1) Nutrient agar medium. </li></ul><ul><li>2) Mackonkey medium. </li></ul><ul><li>B) Stool sample can be cultured on two media: </li></ul><ul><li>1) SS agar medium. </li></ul><ul><li>2) Mackonky medium. </li></ul>
  10. 10. c) Any of other samples can be cultured on the following 3 media : 1) Blood agar medium. 2) Chocolate agar medium. 3) Mackonkey medium.
  11. 11. 1) Sample collection ( from the part where the infection occur) in a sterile container. 2) Pour the suitable previously media in sterile Petri dishes. Example : If the sample is stool, SS agar medium and Mackonkey medium should be poured in Petri dishes. 3) Leave the dishes for 10 minutes to make the media solidified. Steps of culturing
  12. 12. 4) Immerse a sterile swab in the container which containing the specimen, then roll the swab on the surface of a solid medium And spread the specimen in a corner of the dish by the swab, pull the specimen in 3 lines on the surface of the medium by the swab, then pull the 3 lines into another 3 lines (perpendicular to the previous 3 lines), finally draw a zigzag line by the swab on the medium( See Fig.1) . 5) Incubate the cultures at 37 C° for 24-48 hours in an incubator. 6) Observe the results :
  13. 13. a) If the growth occur on all media; hence the sample was infected with a gram positive bacteria. b) If the growth occur on Mackonkey medium only; hence the sample was infected with a gram negative bacteria . Notes : 1) All the above steps must be performed under sterile conditions ( near to the flame). 2) Don't pour the medium in the dish until cooling and before solidification. 3) Don't speak during the culture.
  14. 14. Figure 1 : Fourth-way streak method
  15. 15. <ul><li>It is ordered to enable the doctor choosing the most </li></ul><ul><li>effective Antibiotic against the micro-organism that </li></ul><ul><li>cause the infection with consideration of : </li></ul><ul><li>a) The case of the patient. </li></ul><ul><li>b) The age of the patient. </li></ul>Antibiotics sensitivity testing Procedure : 1) Take a loop of one colony which previously grown on Mackonkey medium or on all media ( blood agar, chocolate agar or SS agar media) by using a sterile swab.
  16. 16. 2) Then by the swab, spread the loop on a nutrient agar medium in 3 direction to ensure confluence 3) By using a dispenser, antibiotic-impregnated disks are placed onto agar surface ( See fig. 2). 4) As the bacteria on the lawn grow, they are inhibited to varying degrees by the antibiotic diffusion from the disk. 5) It has been determined that zones of inhibition of a certain diameter ( varies for antibiotic and to a lesser extent, bacterial species) correlate with sensitivity or resistance to the antibiotic tested.
  17. 17. Figure 2
  18. 18. 6) Incubate the culture at 37C° for 24 hour in an incubator. 7) Observe the results. Note : the larger the inhibition zone, the more the sensitivity the antibiotic (See fig. 3) Figure 3

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