Medical Microbiology Microscopic slides and media

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1 Medical Microbiology Microscopic slides and media Head of Microbiology Department and Laboratory Medical Immunology : Janina Grzegorczyk MD, PhD, professor Implementators: Małgorzata Brauncajs MD Zbigniew Krzemiński MD, PhD, professor

2 Class 1. General microbiology Morphology of bacterial cell, staining procedures for bacteria, culture media for bacteria, cultivation and isolation techniques for bacteria, preparation of pure cultures. Loeffler medium (slant for the detection of corynebacteria) Enriched media Loewenstein-Jensen medium (slant for the detection of mycobacteria)

3 Class 1. General microbiology Morphology of bacterial cell, staining procedures for bacteria, culture media for bacteria, cultivation and isolation techniques for bacteria, preparation of pure cultures. The pigment does not diffuse into the culture medium (Micrococcus sp., Staphylococcus sp.) Chromogenous bacterial strains The pigment diffuses into the culture medium (Pseudomonas sp.) 3

4 Class 1. General microbiology Morphology of bacterial cell, staining procedures for bacteria, culture media for bacteria, cultivation and isolation techniques for bacteria, preparation of pure cultures. Negatively stained Bacillus sp. cylindrical shape Microscopic slides negative stain Negatively stained cocci spherical shape 4

5 Class 1. General microbiology Morphology of bacterial cell, staining procedures for bacteria, culture media for bacteria, cultivation and isolation techniques for bacteria, preparation of pure cultures. Microscopic slide capsule Positive-negative stain 5

6 Class 1. General microbiology Morphology of bacterial cell, staining procedures for bacteria, culture media for bacteria, cultivation and isolation techniques for bacteria, preparation of pure cultures. A stained preparation of Bacillus subtilis showing endospores as uncoloured and the vegetative cell as violet (Gram endospore stain) Microscopic slides endospores A stained preparation of Bacillus subtilis showing endospores as green and the vegetative cell as red (Schaeffer-Fulton endospore stain) 6

7 Class 1. General microbiology Morphology of bacterial cell, staining procedures for bacteria, culture media for bacteria, cultivation and isolation techniques for bacteria, preparation of pure cultures. Microscopic slides flagella 7

8 Class 3. General microbiology Identification and classification of bacteria according to their biochemical and antigenic properties, determination of bacterial resistance to the drugs. Identification of bacteria based on its biochemical properties API test 8

9 Class 3. General microbiology Identification and classification of bacteria according to their biochemical and antigenic properties, determination of bacterial resistance to the drugs. α-hemolysis (incomplete/green hue) Two types of hemolysis β-hemolysis (complete) 9

10 Class 3. General microbiology Identification and classification of bacteria according to their biochemical and antigenic properties, determination of bacterial resistance to the drugs. Disc diffusion test 10

11 Class 3. General microbiology Identification and classification of bacteria according to their biochemical and antigenic properties, determination of bacterial resistance to the drugs. Etest, Epsilometer test (MIC minimal inhibitory concentration); MIC = 0,125 11

12 Class 3. General microbiology Identification and classification of bacteria according to their biochemical and antigenic properties, determination of bacterial resistance to the drugs. MRSA Methicillin-resistant Staphylococcus aureus Determination of S. aureus sensitivity to methicillin with cefoxitin disc Determination of S. aureus sensitivity review method 12

13 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). Identification of staphylococci in selective-differential medium mannitol salt agar (Chapman s medium). Mannitol-positive staphylococci (S.aureus) a change in the pigmentation of the medium from pink to yellow can be seen; mannitol-negative staphylococci (S.epidermidis) no change in the pigmentation 13

14 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). Rapid distinction between micrococci and staphylococci with furazolidone disc: micrococci on the left side (resistant), staphylococci on the right side (susceptible) 14

15 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). Staphylococcus aureus Gram stain (Gram-positive cocci forming structures similar to wine grapes) 15

16 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). The bacitracin (SP disc) test for identifying Streptococcus pyogenes 16

17 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). The optochin (OP disc) test for identifying Streptococcus pneumoniae 17

18 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). Streptococcus pyogenes Gram stain (Gram-positive cocci forming long chains) 18

19 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). Streptococcus pneumoniae in tissue Gram stain (Gram-positive cocci forming diplococci with capsule) 19

20 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). Enterococcus faecalis Coccosel-agar medium. 20

21 Class 5. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; Gram-positive facultative cocci (genus Staphylococcus, Streptococcus and Enterococcus). Enterococcus faecalis Gram stain (Gram-positive cocci forming pairs or short chains) 21

22 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). Neisseria gonorrhoeae Gram-stained urethral discharge 22

23 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). Treponema pallidum Giemsa stain (spiral shape) 23

24 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). C.diphtheriae Neisser stain (violet storage cells) C.diphtheriae Gram stain (Gram-positive, highly pleomorphic organisms with no particular arrangement) 24

25 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). C.pseudodiphteriticum Neisser stain (without storage cells) C.pseudodiphteriticum Gram stain (Grampositive rods, frequently show club-shaped swellings and palisade arrangement) 25

26 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). Actinomyces israelii Gram stain (Gram-positive bacteria that form long, branching filaments resembling the fungi) 26

27 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). Growth of the species belonging to Mycobacterium in the Loewenstein-Jensen medium: (test tube nr 1 Nocardia spp. agar slant; test tubes nr 2 and 3 mycobacteria in the L-J medium) 27

28 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). Bogen s test 28

29 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). Biochemical tests for mycobacteria (test tube nr 1 niacin test, nr 2 Nocardia spp. slant, test tube nr 3 Bogen s test) 29

30 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). Niacin test 30

31 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). Mycobacterium tuberculosis Ziehl-Neelsen stain: (acid-fast, red-stained mycobacteria) 31

32 Class 6. Oral microbiology Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema). The antimicrobial susceptibility of Mycobacterium tuberculosis H37Rv 32

33 Class 8. Oral microbiology Bacteriology of dental caries and periodontal diseases. Bacteroides sp. (Gram-negative bacteria,short, pleomorphic rods) Anaerobic Gram-negative bacteria Gram stain. Fusobacterium sp. ( Gram-negative bacteria, thin, long, usually sharp-ended rods) 33

34 Class 8. Oral microbiology Bacteriology of dental caries and periodontal diseases. Anaerobic Gram-positive bacteria Gram stain Propionibacterium sp. (Gram-positive bacteria, cylindrical, pleomorphic cells) 34

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