Medical Microbiology Microscopic slides and media

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
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
of corynebacteria)
Loewenstein-Jensen medium (slant for
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
culture medium (Micrococcus sp.,
Staphylococcus sp.)
3
The pigment diffuses into the culture
medium (Pseudomonas sp.)
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
shape
4
Negatively stained cocci – spherical shape
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
endospores as uncoloured and the vegetative cell as
violet – (Gram endospore stain)
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A stained preparation of Bacillus subtilis showing
endospores as green and the vegetative cell as red –
(Schaeffer-Fulton endospore stain)
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
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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)
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β-hemolysis (complete)
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
methicillin with cefoxitin disc
12
Determination of S. aureus sensitivity – review
method
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)
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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)
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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), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative 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), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative 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), Grampositive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gramnegative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema).
C.diphtheriae – Neisser stain (violet storage cells)
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C.diphtheriae – Gram stain (Gram-positive, highly
pleomorphic organisms with no particular
arrangement)
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)
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C.pseudodiphteriticum – Gram stain (Grampositive rods, frequently show club-shaped
swellings and palisade arrangement)
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
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
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
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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)
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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
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
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
Class 8. Oral microbiology – Bacteriology of dental caries and periodontal
diseases.
Anaerobic Gram-negative bacteria – Gram stain.
Bacteroides sp. (Gram-negative bacteria,short,
pleomorphic rods)
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Fusobacterium sp. ( Gram-negative bacteria, thin,
long, usually sharp-ended rods)
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)
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