All media must be sterile & the basic conditions for autoclaving
This presentation is the property of its rightful owner.
Sponsored Links
1 / 50

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


  • 87 Views
  • Uploaded on
  • Presentation posted in: General

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

Download Presentation

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

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


All media must be sterile the basic conditions for autoclaving

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

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


All media must be sterile the basic conditions for autoclaving

  • 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 )


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

N.B + 1-5 – 2 % Agar

N.B + 10-15 % Gelatin


All media must be sterile the basic conditions for autoclaving

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


All media must be sterile the basic conditions for autoclaving

Broth media + Sugars

(Glucose & Galactose & Lactose & Mannose & Maltose)

+

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

+ Durham's tube (Gas indicator)


All media must be sterile the basic conditions for autoclaving

Cooked meat for Anaerobic ONLY

Glutathione


All media must be sterile the basic conditions for autoclaving

Characteristic media

Used in

Identification of Enteric organisms


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • 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.


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • Hemolysin Production

  • Test the ability of the organism to produce:

  • ExoenzymeHemolysin which has destructive effect on the blood cells


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • Sugar (CHO) Fermentation

  • Test the ability of the organism to produce:

  • Acid or Acid & Gas upon sugar fermentation


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

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


All media must be sterile the basic conditions for autoclaving

  • 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.


Mr test

MR Test


Vp test

VP Test


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • 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)


All media must be sterile the basic conditions for autoclaving

  • 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+)


All media must be sterile the basic conditions for autoclaving

  • 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


All media must be sterile the basic conditions for autoclaving

  • 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)


  • Login