Medical Microbiology Microscopic slides and media Head of Micro - PowerPoint PPT Presentation

Medical Microbiology Microscopic slides and media Head of Micro robio biolog logy y Depar artm tment nt and d Laborat ratory ry Medic ical al Immuno munolo logy : Janina ina Grzegorc rczyk yk MD, PhD, , professor Implem plementato ntators rs: Małgorzata Braun aunca cajs js MD MD Zbigniew Krzemiński MD, PhD, , professor

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. Enriched media Loeffler medium (slant for the detection Loewenstein-Jensen medium (slant for of corynebacteria) the detection of mycobacteria)

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. Chromogenous bacterial strains The pigment does not diffuse into the The pigment diffuses into the culture culture medium ( Micrococcus sp., medium ( Pseudomonas sp.) Staphylococcus sp.) 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. Microscopic slides – negative stain Negatively stained Bacillus sp. – cylindrical Negatively stained cocci – spherical shape shape 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. Microscopic slide – capsule Positive-negative stain 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 slides – endospores A stained preparation of Bacillus subtilis showing A stained preparation of Bacillus subtilis showing endospores as uncoloured and the vegetative cell as endospores as green and the vegetative cell as red – violet – (Gram endospore stain) (Schaeffer-Fulton endospore stain) 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. Microscopic slides – flagella 7

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

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

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

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

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 Determination of S. aureus sensitivity – review methicillin with cefoxitin disc method 12

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

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

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

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

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

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

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

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

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

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

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

Class 6 . Oral microbiology – Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Gram- positive non-sporulating bacilli ( genus Corynebacterium, Actinomyces), Gram- negative 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

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

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

Class 6 . Oral microbiology – Bacteria causing inflammations and abscesses within the oral cavity; acid-fast bacteria (genus Mycobacterium), Gram- positive non-sporulating bacilli ( genus Corynebacterium, Actinomyces), Gram- negative 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