Medical Microbiology Culture Media :

Transcription

1 Lecture 3 Dr. Ismail I. Daood Medical Microbiology Culture Media : Culture media are used for recognition and identification (diagnosis) of microorganisms. The media are contained in plates (Petri dishes), in test tubes, flasks or screw capped bottles. Used of media as : a- Simple or basal e.g., Nutrient agar, Nutrient broth. b- Special-purpose media e.g., selective media, enriched media. Types of culture media : 1- Liquid (fluid) media e.g., Nutrient broth, Peptone water. 2- Solid media, e.g., Nutrient agar, MacConkey agar. 3- Semisolid media : e.g., motility media. ÐÒ- -

2 Solid media as agar in special purpose as : 1. Enriched media, simple media enriched with substances e.g., added blood 5-10% added glucose 1-2%. 2. selective media containing inhibitory substance as : e.g., bile salts, antibiotic, dyes,.etc., which favors the growth of the concerned microorganism and inhibit the growth of others, e.g., MacConkey agar, Bismuth Sulphate agar or SS agar. 3. Differential media, certain species produce characteristic growth that can easily recognized or can produce certain effects in the media, e.g., Triple sugar Iron agar (TSI), hemolytic and non-hemolytic species on blood agar. Bacterial Growth Curve : During typical bacteria growth (growth cycle) bacteria cell divide by binary fission and their mass and number increase in an exponential manners. Bacterial growth in culture can be separated into at least four distinct phases : 1. Lag phase : This is period of intense physiologic adjustment involving the induction of new enzymes and the synthesis and assembly of ribosome. There would be increase in the size without any multiplication for same time. In lag phase and during this phase there occur : 1. increase in size of cells 2. increase in metabolic rate 3. adaptation to new environment and necessary enzymes. The length of lag phase depend upon a. type of bacteria b. better the medium, shorter the lag phase. c. The phase of culture from which inoculation in taken ÐÓ - -

3 d. Size or volume of inoculums e. Environmental factors like temperature. 2. Logarithmic (Expontial) phase : This phase is characterized by maximal rate of cell diction and followed by multiplication and increase in number of bacteria. The generation time (i.e., the time required for doubling the number of bacteria cell). During this phase is constant for given bacterial species grown under the same of condition but varies among species for example : the G.T. of Pseudomonas may be as brief as 14 minutes. The G.T. of M. tuberculosis as long 24 hours. In logarithmic phase the bacterial cell start dividing and their number increase by geometric progression with time and during this periods. a. bacteria have high rate of metabolism b. bacteria are more sensitive to antibiotics c. rate of penetration of the medium it depends on the concentration of material in the media. 3. Stationary phase : After some time growth rate becomes stationary and during this period there is a balance between cell growth and division and cell death. In stationary phase after some time a stage comes when rate of multiplication and death becomes almost equal it may be due to : a. depletion of nutrient b. accumulation of toxic products and sporulation may occur during this stage. Bacillus and Clostridium are capable of undergoing sporulation. 4. Decline or death phase : During this phase bacterial cells undergo lyses which reduces the number of viable cells. In decline (death) phase, during this phase ÐÔ - -

4 population decreases due to death of cells the factors responsible for are : a. nutritional exhaustion b. toxic accumulation c. autolysis enzymes. Evolution is common in phase of decline. Counting of bacteria of different period after in occultation and then events of sequences are represented on a graph is called "Growth Curve". See the figure of growth curve below : ÐÕ- -

5 Bacterial Classification : The general classification of the bacterium isolated from an infected patients. Readily observable properties usually are the basis for the grouping into large groups according : 1. Staining properties : a. Gram ( + ve ) bacteria e.g. Staphylococcus, Streptococcus... b. Gram ( - ve ) bacteria e.g. E. coli, Salmonella. c. Acid-fast stain e.g., Myco. Tuberculosis. ÐÖ- -

6 Fig. Bacterial cell wall of gram ( + ve ), up and gram ( - ve ), dawn 2. Morphological features : a. Shape e.g., Spherical ( cocci ), Bacilli, Coccobacilli b. Arrangement : as e.g., single cell cocci two Diplococci or more as Streptococcus. 3. Metabolic activity : a. Oxygen (O 2 ) requirements : e.g. Aerobes, as Vibrio cholera, An aerobes as Clostridium,Facultative as Enterobacter, Microaerophilic and Aerotolerent. b. metabolic carbohydrate utilization : e.g. fermented lactose as E. coli, Klebsiellia or non fermented lactose as : Salmonella, Shigella. c. Enzyme production : a. Oxidase e.g., Staphylococcus, Moraxella b. Catalase as Moraxella. 4. Serologic reactivity as genus specific - species-specific - specific for subgroup (serotype). 5. Genetic Classify on base of genetic a. ability to exchange genetic information as : transformation, conjugation. Ð - -

7 b. nucleotide base composition % G=C, A=T. c. nucleic acid homology hybridization study. d. nucleic acid sequences. Glossary Allergy : Hypersensitivity especially of the IgE mediated type mediated type Antibody: A protein produced by the body response to a foreign substance that reacts specifically with that substance. Antigen : A substance that can cause the production of specific antibodies and combine with those antibodies. Envelop : The structure which surround the cytoplasm of the cell. It includes the capsule cell wall, and cytoplasmic membrane. Colony: A population of bacterial cells arising from a single cell. Colonization: Establishment of a site of reproduction of microbes on a material animal or person without necessarily resulting in tissue invasion or damage. Spike : A protein on the surface of a virus that is important in attachment to the host cell. Spore : A specialized reproductive cell. Toxic : Poisonous. Toxin : A poisonous chemical substance. - - ÑÎ