Methods of Classification and identifying microorganisms

Morphology a branch of bioscience dealing with the study of the form and structure of organisms and their specific structural features includes aspects of the outward appearance as well as the form and structure of the internal parts Identification Identification is the determination of whether a microorganism should be placed within a group of microorganisms known to fit within some classification scheme. Identification techniques: different criteria may be employed for identification Techniques include: Morphological identification Differential staining Use of differential media Serological methods Flow cytometry Phage typing Protein analysis Comparisons of nucleotide sequences Morphological identification A number of morphological characteristics are useful in bacterial identification. Eg. The presence or absence of endospores, flagella, glycocalyx Additional considerations include colony morphology, cell shape, cell size Gram Stain Used to identify the cause of an infection. A negative Gram stain is often reported as "no organism seen." Which means there were no or not enough microorganisms present in the sample. Positive Gram stain results usually include a description of what was seen on the slide. This typically includes whether the bacteria are Gram-positive (purple) or Gram-negative (pink) as well as their shape and Sometimes size, relative quantity, and grouping of the bacteria Acid-fast Stain Bacteria with an acid-fast cell wall when stained by the acid-fast procedure, resist decolorization with acid-alcohol and stain red, the color of the initial stain. The genus Mycobacterium and the genus Nocardia are acid-fast. All other bacteria will be decolorized and stain blue. Negative capsule Stain Identify presence of capsules. The background is stained using an acidic stain and the cell itself stained using a basic stain. Spore Stain Identify if there are spores. Positive results are that Spores will be a light green and Vegetative cell walls will pick up the counterstain safranin.

Flagella Stain To stain bacterial flagella, thus to reveal the presence or absence of flagella as well as their arrangement on the perimeter of the cell. Results: Bacteria and flagella will appear golden brown. Excess stain is often observed on the slides and illustrates the necessity of beginning the procedure with thoroughly cleaned microscope slides. Starch Hydrolysis Test Starch agar is a differential medium that tests the ability of an organism to produce certain exoenzymes which some bacteria secrete to degrade starch into subunits that can then be utilized by the organism. Iodine is added after incubation and it turns blue-black in the presence of starch. A clearing around the bacterial growth indicates that the organism has hydrolyzed starch. Coagulase Test Used to differentiate Staphylococcus aureus from coagulase-negative staphylococci. If positive: Macroscopic clumping would be observed in the plasma within 10 seconds and no clumping in the saline drop. If negative: No clumping will be observed. Oxidase Test a test used in microbiology to determine if a bacterium produces certain cytochrome c oxidases. A positive test (OX+) will result in a color change to pink, through maroon and into black, within 1030 seconds. A negative test (OX-) will result in a light-pink or absence of coloration. Citrate Test Detects the ability of an organism to use citrate as the sole source of carbon and energy. Positive: Growth on the medium even without colour change will be considered as positive. Negative: No growth observed. Serological methods Employ antibodies and include agglutination tests, ELISAs, Western blots etc. antibodies are highly selective in terms of the proteins (or other cell structures) to which they bind, to the point that they are able to distinguish the proteins coming from one bacterial species among many species, or even one strain among many strain. Phage typing Bacteriophages (or phages) are viruses that infect bacteria. Phage can be very specific in what bacteria they infect and the pattern of infection by many phages may be employed in phage typing to distinguish bacterial species and strains. Genelogy The study of the patterns of inheritance of specific traits, relating to genes and genetic information. Also known as heredity. Modern theories explain how traits are passed down from parent to offspring. Immunology

A subfield of biology that deals with the study of antigens and the immune process and how humans and higher animals fight off disease.