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Theme J03 Theme J03: Cultivation Task 1: Characterisation of media and their production Task 1a: Most important media in medical microbiology Look at given media and write here the type of medium according to explanation given by your teacher. Do not forgot to write, whether the medium is solid or liquid and whether it is in a Petri dish or in a test tube. Name of medium Liquid/solid Colour Type of medium Used for Petri dish/test t. (selective, …) bacteria: Task 1b: Production of blood agar. Look at the video. Describe the procedure on the opposite side of this sheet. Task 2: Influence of physical and chemical conditions on the bacterial growth Task 2a: Influence to oxygen Five strains (J, K, L, M, N) were cultured on agar plates in four types of conditions: a) normal athmosphere b) elevated CO2 concentration c) decreased oxygen conc. d) no oxygen at all Write „growing“, „not growing“, assess, which strain is strictly aerobic, facultatively anaerobic, strictly anaerobic, microaerophilic, capnophilic normal air elevated traces of no oxygen conclusion CO2 oxygen only Task 2b: Influece of salts and antibiotics You can see three strains on various types of media. Describe the presence/absence of growth (GROWS – DOES NOT GROW) Strain Blood agar BA + NaCl BA + NaCl Slanetz BA with (BA) (6,5 %) (10 %) Bartley agar amicacine E – enterococcus SR – streptoccus ST – staphylococ. Name _________________________ Group______ Gen. med. – stomatology 1 Theme J03 Task 2c: Influence of temperature Like the previous, only with the same medium, but different temperature: Strain 4 °C 37 °C 42 °C Task 3: Properties of the two most common diagnostic and selective-diagnostic media Task 3a: Blood agar – viridation and hemolysis Describe haemolytical properties of four strains on blood agar. Read against light. Streptococcus pyogenes Streptococcus agalactiae Streptococcus pneumoniae Enterococcus faecalis Task 3b: Endo agar – presence/absence of growth, lactose fermentation Describe growth/no growth, and changes of the medium surrounding the colonies. Staphylococcus epidermidis Escherichia coli Salmonella Enteritidis Task 4: Description of morphologic characteristics of colonies Describe three strains of bacteria Strain Size Colour Shape Profile Surface Edges Translucency Consistency Odour Surroundings Strain Strain Task 5: Inoculation of samples and streins on solid media Task 5a: Inoculation of a swab Task 5b: Inoculatiob of a strain Inuculate a swab (a) and a strain (b) on the medium. Draw your result to the opposite side of this sheet. Check-up questions: (Use another sheet of paper or another side of this one for answers) 1. Parafin oil is usually poured on VL-broth when used for bacteria cultivation. Why? 2. Why do we add the red blood cells after (and not before) the decrease of temperature of the agar medium? 3. Why, at bacteriological media preparing, agar is used rather then gelose? 4. Microaerophilia and capnophilia: is it the same? 5. Is the ability of staphylococci to grow at high NaCl concentracions connected with its adaptation to the host organism? 6. Which growth characteristics are not recognizable using your eye? And which one needs a loop contact with a colony? 7. Why is it so important to obtain individual colonies when we culture bacteria on solid media? 8. Blood agar is usually made of „basis for blood agar“ (approximatelly, this one is equivalent to nutrient agar) and defibrinated sheep blood. Is it possible to add sheep blod to other bases than the one listed above? Name _________________________ Group______ Gen. med. – stomatology 2