slide1
Download
Skip this Video
Download Presentation
All media must be sterile & the basic conditions for autoclaving 

Loading in 2 Seconds...

play fullscreen
1 / 50

All media must be sterile & the basic conditions for autoclaving  - PowerPoint PPT Presentation


  • 152 Views
  • Uploaded on

All media must be sterile & the basic conditions for autoclaving   Temperature = 121 o C  Pressure = 15 PSI  Time = 20 minutes. Ordinary أكتر ميديا بتستخدم في نمو البكتريا Enriched غنية فيها اضافات البكتريا محتاجاها علشان تنمو وبالذات البكتريا الممرضة Enrichment

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about ' All media must be sterile & the basic conditions for autoclaving ' - evonne


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
slide1

All media must be sterile & the basic conditions for autoclaving 

 Temperature = 121oC  Pressure = 15 PSI  Time = 20 minutes

slide2

Ordinary

  • أكتر ميديا بتستخدم في نمو البكتريا
  • Enriched
  • غنية فيها اضافات البكتريا محتاجاها علشان تنمو وبالذات البكتريا الممرضة
  • Enrichment
  • فيها حاجات تزود العدد بتاع البكتريا اللي موجودة بكمية قليلة علشان أزود عددها عن الـ N. Flora
  • Selenite broth  allow growth of Salmonella & prevent E. Coli
  • Selective
  • فيها مادة تخلي MO واحد يعيش والباقي يموت (تختارة)
  • Differential
  • فيها مادة تخلي شكل الـ MO مختلف عن الـ MO التاني غالباً يكون الاختلاف في اللون
  • Characteristic
  • ميديا بستخدمها في التعرف على البكتريا عن طريق الميتابوليزم بتاعها (Sugar fermentation )
slide7

Cultivation of bacteria

  • meat extract & Pepton &
  • 0.5% NaCl, neutral PH
  • light yellow transparent fluid
  • Indole Production
  • Base for sugar media
  • 1% Pepton + 0.5% NaCl + water
slide8

N.B + 1-5 – 2 % Agar

N.B + 10-15 % Gelatin

slide10

Differentiate M.O according 2 Hemolytic activity

N. Agar  100 C 55 C  5-10 % Sheep or Ox Blood

simmon citrate agar
Simmon Citrate Agar
  • Upon citrate utilization the PH of the media will be increased causing change in color of the media into  blue
  • Due to Bromothymol Blue
  • Ability of MO to use citrate as carbon source for energy.
  • Degrade citrate producing CO2 which react with Na & water forming Na carbonate (alkaline product) which change color or BTB from green into deep prussian blue
slide20

Broth media + Sugars

(Glucose & Galactose & Lactose & Mannose & Maltose)

+

Phenol Red (Yellow in Acidic PH & Purple/Red in Basic PH)

+ Durham\'s tube (Gas indicator)

slide25

Characteristic media

Used in

Identification of Enteric organisms

slide27

Starch Hydrolysis

  • Test the ability of the organism to produce:
  • ExoenzymeAmylase which breaks down the Starch (Complex CHO of large molecule  Cannot pass through the cytoplasmic membrane) into Monosaccharide (MS = Simple can be used by the organism)

 Inoculate the Organism in Starch agar + add I2

 Amylase producing organism is surrounded by a clear zone(MS)

while the remaining of the media will stain with the violet color

slide28

Casein Hydrolysis

  • Test the ability of the organisms to produce:
  • Proteolyticexoenzymes (Proteinase which hydrolyze casein)
  • Casein  Main protein of milk Responsible for the white color of milk.
  • Hydrolysis of casein  Form more soluble & transparentcompounds (peptides &aa)
  • Upon growing the organism on casein media the area surrounding the proteinase producing organism will appear transparent.
  • Casein hydrolysis is called Peptonization or Proteolysis.
slide29

Gelatin Hydrolysis (Liquefaction)

  • Test the ability of the organism to produce:
  • ExoenzymeGelatinsae which liquefy gelatin.
  • Gelatin hydrolysis (Liquefaction) is indicated by:
  • loss in ability to solidify even after refrigeration
slide31

Catalase Production

  • Test the ability of the organism to produce:
  • Catalase enzyme that degradatesH2O2 O2 + H2O + Air bubbles.
  • H2O2 is added to the bacterial media
  • Presence of gas bubblesmeans that the organism produces catalase
slide32

Oxidase Production

  • Test the presence of Cytochrome Cin the respiratory chain.
  • Aerobic organisms with Cytochrome Ccan oxidize amines to form coloredproducts.
  • This Test is specific for Pseudomonas Aeruginosa.
  • Wet F. Paper with
  • 1% N,N,N\',N\' Tetra methyl - P-Phenylene-Diamine (TMPD) (KovacOxidase reagent)
  • allow to dry & Pick bacterial colony with sterile toothpick  add to F. Paper
  • A purple color is produced
slide33

Hemolysin Production

  • Test the ability of the organism to produce:
  • ExoenzymeHemolysin which has destructive effect on the blood cells
slide34

Urease Production

  • Test the ability of the organism to produce:
  • Urease enzyme which splits urea in urea media to form Ammonia + CO2
  • Accumulation of Ammonia will produce alkaline PH  Turns the color of indicator (phenol red) into Pink
slide35

H2S Production

  • H2S from Organic S or Inorganic S
  • Hydrogen Sulfide is detected by iron salt.
  • The presence of black precipitate is indication of H2S production.
  • Inoculate media peptone iron agar or TSI (Na2S2O3)
  • Black color will indicate H2S production
slide36

Sugar (CHO) Fermentation

  • Test the ability of the organism to produce:
  • Acid or Acid & Gas upon sugar fermentation
slide37

Nitrate Reduction

  • Test the ability of the organism to produce:
  • Nitrate reductaseenzyme which can reduce nitrate into nitrite

 Inoculate organism into nitrate broth

 Incubate at 37 C for 48 hrs

 Add 1 ml of coupling reagent

(sulfanilic acid & 1 ml of dimethyl alpha naphthyl amine reagent)

If the organism produce nitrate reductase the nitrate in the media will be reduced into nitrite & Color become red precipitate

slide38

Ammonia Production

  • Test the ability of the organism to:
  • Degradate the organic nitrogen in the protein into ammonia.

 Inoculate organism in 4% peptone water

 Incubate at 37 c for 2, 4, 7 days

 Add Nessler’s reagent.

Appearance of Yellow-Orange or brown color indicates +Ve test

slide39

IMViC tests used for  Identification & Differentiation of Enterobacteriaceae (Klebsiella & Enterobacter & E. Coli) ( All are Lactose Fermenters)

slide40

Indole Test

  • Test the ability of organism to break down tryptophan into indole.
  • Incubate tryptophan (Peptone) broth media with the tested organism.
  • The Presence of indole can be detected by Kovac’s reagent
  • (Para DimethylAminobenzaldehyde in amyl alcohol)

Kovac\'s reagent (yellow color) reacts with indole & produce

(red color) on the surface of the test tube.

slide43

Methyl Red Test

  • Test the ability of organism to ferment the glucose & produce acids which will change the color of M.R (PH indicator) into red color
slide44

VogasProskaurTest

  • Test the ability of organism to ferment the glucose & produce neutral products which will change the color of indicator into Pink color
  • The reagents used for the VP test are
  • Barritt\'s A (Alpha-Napthol) & Barritt\'s B (Potassium-Hydroxide)
slide45

MR & VP tests is done on MR-VP broth media

  • (contains glucose & peptone)
  • MR & VP tests:
    • E. Coli is (MR+/VP-)
    • Klebsiella & Enterobacteraerogenes is (MR-/VP+)
slide46

Citrate Test

  • Test the ability of organism to utilize citrate as its only source of carbon.
  • Simmon’s Citrate media used in this test
  • Bacteria can break citrate into organic acids & CO2 CO2 form a basic compound (Na2CO3)

Adding Bromothymol Blue Detects the presence of Na2CO3 by turning into blue (+Ve test)

eosin methylene blue medium
Eosin-Methylene blue medium
  • Lactose / Esoin & MB
  • Permit differentiation between enteric lactose fermenters and non-fermenters
  • Alos in identification of E. coli
slide48

Lactose fermenter: purple black

  • Non- Lactose fermenter: colorless
  • E.coli: metallic green sheen
motility test
Motility Test
  • The medium contain triphenyltetrazolium which is reduced into red color by the stabbed bacterial growth.
  • Motile bacteria appear as diffused growth (with red color)
  • Non-Motile bacteria appear as single line of growth (the original stabbed line with pin color)
ad