Practical examination
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1 Practical examination
2 I. Sterile media
3 1. Bouillon, 2. Slant agar, tube agar
4 4. Enrichment media: meat bouillon
5 3., 5., 6.: Agar, blood agar and chocolate agar plates
6 7. Selective and differentiating media c. Eosin methylene blue (EMB) Lactose + colonies Lactose - colonies
7 7. Selective and differentiating media g. TCBS Vibrio cholerae
8 8. Transport media Transport swabs (e.g. Stuart) Transport growth media (e.g. Uricult)
9 9. Blood culture
10 10. Virus transport medium
11 II. Cultures
12 Staphylococcus aureus and S. epidermidis on agar plate and blood agar plate
13 4. S. epidermidis (S) and S. saprophyticus (R) with novobiocin disc
14 5. Streptococcus pyogenes on blood agar
15 6. Str. mitis on blood and chocolate plates
16 7. Streptococcus pneumoniae on blood agar plate (optochin S)
17 8. Haemophilus influenzae on chocolate agar
18 9/12. E. coli and Klebsiella on agar plate
19 9/12. E. coli and Klebsiella on EMB plate
20 10. Proteus on agar and blood agar (swarming!)
21 11. Pseudomonas on agar plate
22 13. Salmonella on bismuth sulphite medium
23 14. E. coli and Salmonella on brillant green medium
24 15. E. coli and Shigella on DC medium
25 16. Faeces of patient with dysentery on DC medium (E. coli + Shigella)
26 17. Russel medium Glucose (or dextrose), saccharose (or sucrose), lactose fermentation of lactose: the whole medium turns deep red fermentation of other sugars only: little red explosion due to gas formation
27 18. Urease test NH 2 -CO-NH 2 (ureum) CO 2 + NH 3 (ammonia) Christensen medium: indicator (phenol red) urease +: purple urease -: citric yellow Urease +: Proteus, Klebsiella, Helicobacter
28 19. Corynebacterium on Clauberg and Löffler medium No picture Clauberg Löffler
29 20. Mycobacterium tuberculosis on Löwenstein-Jensen medium
30 21. Leptospira on Korthof medium
31 22. Bacillus cereus on agar plate
32 Bacillus cereus on blood agar
33 Clostridium tetani and C. perfringens on Holman and thioglycolate media
34 GasPak
35 III. Reactions and methods
36 1. Elek test Aim: test for toxin production Filter paper with antitoxin Precipitation arch Corynebacterium
37 2. Widal-type reaction Gruber-Widal: abdominal typhus (Salmonella) Weil-Felix: epidemic typhus (Rickettsia) Wright: Brucella / Francisella With Proteus OX19 antigen! titre: the smallest amount of antibody, which can cause positive reaction!!!
38 3. Wasserman test for syphilis (CF) Positivity: no hemolysis 1 2 2: 4-fold increase in titre compared to 1 - indicates infection
39 4. Hemagglutination (HA) and -inhibition (HAI) hemagglutination hemagglutination inhibition CF test hemolysis (-) no hemolysis (+)
40 8. Phage typing
41 10. Steps of Gram staining 1/a. Crystal violet 2 1/b. Lugol solution 1 2. Differentiation with 96 % ethanol 3. Washing with water 4. Safranine 1 5. Washing 6. Drying 7. Microscopic examination (immersion)
42 11. Steps of Ziehl-Neelsen staining acid fast staining Carbol-fuchsin (on filter paper!) heating 3x (until steaming) Differentiation: acidic alcohol 3% HCl, 96% ethanol acid fast thorough water washing counter staining 1-2 methylene blue or malachite green careful rinsing, drying with filter paper
43 Antibiotic susceptibility test with disc diffusion ( antibiogram )
44 13. Commercial kits for identification
45 IV. Microscopic preparations
46 1.a Staphylococcus (Gram +)
47 1.b Streptococcus pyogenes (Gram +)
48 1.c Streptococcus pneumoniae (G +)
49 1.d Neisseria (Gram -)
50 1.e E. coli (Gram -)
51 1.f Haemophilus influenzae (Gram -)
52 1.g Bacillus (Gram + spore forming)
53 1.h Clostridium tetani and perfringens (Gram + spore forming)
54 1.i Corynebacterium - Neisser staining
55 2.a Neisseria gonorrheae in pus
56 2.b Candida albicans from culture
57 3. Silver impregnation: Leptospira
58 4. Ziehl-Neelsen staining: Mycobacterium tuberculosis
59 5.a Plasmodium falciparum in thin blood film (Giemsa staining)
60 5.b Trypanosoma brucei in thin blood film (Giemsa staining)
61 6. Anorectal cellulose-tape preparation (Enterobius)
62 VI. Tests performed by the students
63 Catalase test Catalase +: Staphylococci Catalase -: Streptococci performance: adding a few drops of H 2 O 2 to the culture, or smear bacteria into a drop of H 2 O 2 on slide: strong bubbling if positive - + H 2 O 2 H 2 O + O 2
64 Clumping test - Coagulase test Coagulase +: Staphylococcus aureus Coagulase -: all other Staphylococci ( CNS ) Staphaurex : latex agglutination performance: mixing bacteria with fibrinogenbound latex particles on microscope slide clumping
65 Oxidase test Oxidase +: Pseudomonas, Neisseria, Vibrio, Campylobacter purpose: detection of cytochrome-oxidase enzyme diagnostic use: differentiation of obligate aerobes performance: filter paper on microscope slide add oxidase reagent + - (= parephenylene-diamine derivative) add bacteria positivity: pink / purple colour
66 Indol test triptophane indol (triptophanase enzyme) Indol +: E. coli amylalcoholic (=organic) reagent indol is dissolved here after mixing intensive red ring on top (alcohol is lighter then water!) can be performed in ureumindol tube
67 6. Slide agglutination with E. coli Bacterium suspension (control) NEGATIVE POSITIVE White particles, clear solution
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