Diagnosis of Bacterial InfectionsDiagnosis of Bacterial Infections
PRESENTER
Shakira
Ghazanfar
PhDScholar
24-1-2013
ISLAMA...
Diagnosis of BacterialDiagnosis of Bacterial
InfectionsInfections
TYPES OF SAMPLESTYPES OF SAMPLES
EXAMINEDEXAMINED
•Animal Tissues – Blood samples
•Foods of animal origin
•Meat and meat p...
Steps in diagnostic isolation andSteps in diagnostic isolation and
identification of bacteriaidentification of bacteria
 ...
IdentificationIdentification
 Colony CharacterColony Character
 Stainining ReactionStainining Reaction
 MorophologyMoro...
THE GRAM STAINTHE GRAM STAIN
 A colony is dried on a slideA colony is dried on a slide
 Step 1. Staining with crystal vi...
Crystal
violet
Gram's
iodine
Decolorise with
acetone
Counterstain with
e.g. methyl red
Gram-positives
appear purple
Gram-n...
Gram-positive rods
Gram-negative rods
Gram-positive cocci
Gram-negative cocci
Gram-positive
cocci
Anaerobic
Gram-positive rods
Anaerobic
Gram-negative rods
Anaerobic
Gram-positive cocci
Anaerobic
Gram...
Gram-negativeGram-negative
Pink on
Macconkey
Agar
Pale on
Macconkey
Agar
Biochemical TestsBiochemical Tests
VocabularyVocabulary
AerobicAerobic – required oxygen.– required oxygen.
AnaerobicAnaerobic – growing only in the absence ...
Media and CultureMedia and Culture
•Media: Nutrients (agar, pH indicators, proteins
and carbohydrates) used to grow organi...
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Bacterial infection (by shakira)

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  • Here is photograph of colonies growing on SBA. This is the vaccine strain of Bacillus anthracis , but non-vaccine strains look substantially similar. You can see the flat, wavy-edged, ground-glass appearing colonies.
  • Bacterial infection (by shakira)

    1. 1. Diagnosis of Bacterial InfectionsDiagnosis of Bacterial Infections PRESENTER Shakira Ghazanfar PhDScholar 24-1-2013 ISLAMABAD ‫الرحيم‬ ‫الرحمن‬ ‫اللة‬ ‫بسم‬‫الرحيم‬ ‫الرحمن‬ ‫اللة‬ ‫بسم‬
    2. 2. Diagnosis of BacterialDiagnosis of Bacterial InfectionsInfections
    3. 3. TYPES OF SAMPLESTYPES OF SAMPLES EXAMINEDEXAMINED •Animal Tissues – Blood samples •Foods of animal origin •Meat and meat products •Milk and milk products •Fish and fish products •Water •Animal feeds
    4. 4. Steps in diagnostic isolation andSteps in diagnostic isolation and identification of bacteriaidentification of bacteria  Step 1. Samples of body fluids . blood, urine,Step 1. Samples of body fluids . blood, urine, cerebrospinal fluid, Abscess)cerebrospinal fluid, Abscess)  streaked on culture platesstreaked on culture plates  isolated colonies of bacteria appear afterisolated colonies of bacteria appear after incubation for one to several days.incubation for one to several days.  Observation of colonies (size, texture, color, &Observation of colonies (size, texture, color, & hemolysis)hemolysis)  differentiating characteristic- e.g oxygendifferentiating characteristic- e.g oxygen requirementrequirement
    5. 5. IdentificationIdentification  Colony CharacterColony Character  Stainining ReactionStainining Reaction  MorophologyMorophology  Biochemical tests and serological testsBiochemical tests and serological tests -Commercial kits commonly used, e.g. API20-Commercial kits commonly used, e.g. API20
    6. 6. THE GRAM STAINTHE GRAM STAIN  A colony is dried on a slideA colony is dried on a slide  Step 1. Staining with crystal violet.Step 1. Staining with crystal violet.  Step 2. Fixation with iodine stabilizes crystalStep 2. Fixation with iodine stabilizes crystal violet staining. All bacteria remain purple orviolet staining. All bacteria remain purple or blue.blue.  Step 3. Extraction with alcohol or other solvent.Step 3. Extraction with alcohol or other solvent. Decolorizes some bacteria (Gram negative) andDecolorizes some bacteria (Gram negative) and not others (Gram positive).not others (Gram positive).  Step 4. Counterstaining with safraninStep 4. Counterstaining with safranin
    7. 7. Crystal violet Gram's iodine Decolorise with acetone Counterstain with e.g. methyl red Gram-positives appear purple Gram-negatives appear pink The Gram Stain
    8. 8. Gram-positive rods Gram-negative rods Gram-positive cocci Gram-negative cocci
    9. 9. Gram-positive cocci Anaerobic Gram-positive rods Anaerobic Gram-negative rods Anaerobic Gram-positive cocci Anaerobic Gram-negative cocci
    10. 10. Gram-negativeGram-negative Pink on Macconkey Agar Pale on Macconkey Agar
    11. 11. Biochemical TestsBiochemical Tests
    12. 12. VocabularyVocabulary AerobicAerobic – required oxygen.– required oxygen. AnaerobicAnaerobic – growing only in the absence of– growing only in the absence of oxygen.oxygen. Antibiotic susceptibility testingAntibiotic susceptibility testing –– determining the susceptibility ofdetermining the susceptibility of microorganisms to specific antibiotics.microorganisms to specific antibiotics. BacillusBacillus – rod-shaped bacterium– rod-shaped bacterium CoccusCoccus – spherical bacterium– spherical bacterium ColonyColony - defined mass of bacteria assumed to- defined mass of bacteria assumed to assumed to have grown form a singleassumed to have grown form a single organismorganism CommunicableCommunicable – able to be transmitted– able to be transmitted directly of indirectly from one individualdirectly of indirectly from one individual to another.to another. CultureCulture – growth of microorganisms in a– growth of microorganisms in a special medium; the process of growingspecial medium; the process of growing microorganisms in the laboratory.microorganisms in the laboratory. DNADNA – nucleic acid found primarily in the– nucleic acid found primarily in the nucleus of all living cells that carriesnucleus of all living cells that carries genetic information; deoxyribonucleicgenetic information; deoxyribonucleic acid.acid. Fastidious organismFastidious organism – organism that requires– organism that requires special nutritional factors to survive.special nutritional factors to survive. FissionFission – asexual reproduction of a– asexual reproduction of a microorganism.microorganism. FormalinFormalin – solution of formaldehyde used as a– solution of formaldehyde used as a fixative of preservative.fixative of preservative. Gram-negativeGram-negative – designation for bacteria that– designation for bacteria that lose the crystal violet (purple stain) andlose the crystal violet (purple stain) and retain the safranin (red stain) in the Gramretain the safranin (red stain) in the Gram stain procedure.stain procedure. Gram-positiveGram-positive – designation for bacteria that– designation for bacteria that retain the crystal violet( purple stain) in theretain the crystal violet( purple stain) in the Gram stain procedure.Gram stain procedure. Gram stainGram stain – differential stain used to classify– differential stain used to classify bacteria.bacteria. HostHost – organism from which a parasite obtains– organism from which a parasite obtains nutrients and in which some or part of thenutrients and in which some or part of the parasites' life cycle is carried.parasites' life cycle is carried. HyphaeHyphae – filaments of mold that makes up the– filaments of mold that makes up the mycelium.mycelium.
    13. 13. Media and CultureMedia and Culture •Media: Nutrients (agar, pH indicators, proteins and carbohydrates) used to grow organisms outside of their natural habitats •Culture: The propagation of microorganisms using various media
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