微生物学课件：10 微生物鉴定.ppt

Chapter 10The Identification of Microorganisms 1 Learning objectives for this sectionFollowing completion of this section you should be able to;Determine the appropriate methods for identification of different MOsDifferentiate between culture methods and biochemical methods of MO identificationUnderstand some of the rapid methods used for MO detection Identification of MicrobesMaintain pure cultureAscertain colony morphologyStaining techniques – cell shape, arrangements and type of cell wallUse of Selective and Differential mediaBiochemical and Serological tests - API kits / Enterotube and BiologRapid methods Identification of MicrobesInitial steps involve establishment of pure culture (single species):Streak platePour platePick isolates (individual colonies) from plates and prepare pure culture by e.g. streaking onto new plate or inoculating sterile broth. Colony MorphologyDifferent types of bacteria have different appearances or “morphology” on different agar.Can distinguish different morphologies based on shape, size, elevation, colour, opacity, edge pattern, surface etcEg: Micrococcus spp on nutrient agar is yellow, circular, smooth edge with convex elevation.Eg: Bacillus spp often has irregular shaped, beige(浅褐色）, lobate（叶状） or curled（卷发状）, slimy colonies on nutrient agar. Micrococcus leuteusBacillus cereusDifferentiation between two bacteria Colony Morphology Staining TechniquesLiving bacteria almost colourlessBacteria are chemically different to their surroundings and therefore react differently to stains than background, making them distinguishable and visible.Commonly used stains include:Basic stainGram stain,Acid-fast stainStructural stains: Endospore, capsule stain Direct staining with Basic dyesBacterial cell mem normally negatively charged in neutral pH environmentTherefore attract positively charged ions, such as those in dyes E.g. methylene blue - +ion.Fix smear of bacterial culture on slideBasic dyes (where colour is in +ve ion of the stain) incl