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 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.)

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 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

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 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

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 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)

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 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

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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.

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) β-hemolysis (complete)

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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.

Disc diffusion 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.

Etest, Epsilometer test (MIC – minimal inhibitory concentration); MIC = 0,125

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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.

MRSA – Methicillin-resistant Staphylococcus aureus

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

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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).

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

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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).

Rapid distinction between micrococci and staphylococci with furazolidone disc:

micrococci on the left side (resistant), staphylococci on the right side (susceptible)

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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).

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

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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 optochin (OP disc) test for identifying Streptococcus pneumoniae

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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 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)

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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).

Enterococcus faecalis – Coccosel-agar medium.

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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).

Enterococcus faecalis – Gram stain (Gram-positive cocci forming pairs or short

chains)

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 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

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 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)

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 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)

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 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)

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 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)

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 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)

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 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).
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), Gram-
positive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gram-
negative 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), Gram-
positive non-sporulating bacilli (genus Corynebacterium, Actinomyces), Gram-
negative aerobic cocci (genus Neisseria) and spirochetes (genus Treponema).
Niacin test

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 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).
Mycobacterium tuberculosis – Ziehl-Neelsen stain: (acid-fast , red-stained
mycobacteria)

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 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).
The antimicrobial susceptibility of Mycobacterium tuberculosis H37Rv

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 Class 8. Oral microbiology – Bacteriology of dental caries and periodontal
diseases.

Anaerobic Gram-negative bacteria – Gram stain.

Bacteroides sp. (Gram-negative bacteria,short, Fusobacterium sp. ( Gram-negative bacteria, thin,
pleomorphic rods) long, usually sharp-ended rods)

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 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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