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Medical MicrobiologyMicroscopic slides and mediaHead of Microbiology Department and Laboratory Medical Immunology : Janina Grzegorczyk MD, PhD, professorImplementators:Małgorzata Brauncajs MDZbigniew Krzemiński MD, PhD, professor

Class 1. General microbiology – Morphology of bacterial cell, stainingprocedures for bacteria, culture media for bacteria, cultivation and isolationtechniques for bacteria, preparation of pure cultures.Enriched mediaLoeffler medium (slant for the detectionof corynebacteria)Loewenstein-Jensen medium (slant forthe detection of mycobacteria)

Class 1. General microbiology – Morphology of bacterial cell, stainingprocedures for bacteria, culture media for bacteria, cultivation and isolationtechniques for bacteria, preparation of pure cultures.Chromogenous bacterial strainsThe pigment does not diffuse into theculture medium (Micrococcus sp.,Staphylococcus sp.)3The pigment diffuses into the culturemedium (Pseudomonas sp.)

Class 1. General microbiology – Morphology of bacterial cell, stainingprocedures for bacteria, culture media for bacteria, cultivation and isolationtechniques for bacteria, preparation of pure cultures.Microscopic slides – negative stainNegatively stained Bacillus sp. – cylindricalshape4Negatively stained cocci – spherical shape

Class 1. General microbiology – Morphology of bacterial cell, stainingprocedures for bacteria, culture media for bacteria, cultivation and isolationtechniques for bacteria, preparation of pure cultures.Microscopic slide – capsulePositive-negative stain5

Class 1. General microbiology – Morphology of bacterial cell, stainingprocedures for bacteria, culture media for bacteria, cultivation and isolationtechniques for bacteria, preparation of pure cultures.Microscopic slides – endosporesA stained preparation of Bacillus subtilis showingendospores as uncoloured and the vegetative cell asviolet – (Gram endospore stain)6A stained preparation of Bacillus subtilis showingendospores as green and the vegetative cell as red –(Schaeffer-Fulton endospore stain)

Class 1. General microbiology – Morphology of bacterial cell, stainingprocedures for bacteria, culture media for bacteria, cultivation and isolationtechniques for bacteria, preparation of pure cultures.Microscopic slides – flagella7

Class 3. General microbiology – Identification and classification of bacteriaaccording to their biochemical and antigenic properties, determination ofbacterial resistance to the drugs.Identification of bacteria based on its biochemical properties – API test8

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

Class 3. General microbiology – Identification and classification of bacteriaaccording to their biochemical and antigenic properties, determination ofbacterial resistance to the drugs.Disc diffusion test10

Class 3. General microbiology – Identification and classification of bacteriaaccording to their biochemical and antigenic properties, determination ofbacterial resistance to the drugs.Etest, Epsilometer test (MIC – minimal inhibitory concentration); MIC 0,12511

Class 3. General microbiology – Identification and classification of bacteriaaccording to their biochemical and antigenic properties, determination ofbacterial resistance to the drugs.MRSA – Methicillin-resistant Staphylococcus aureusDetermination of S. aureus sensitivity tomethicillin with cefoxitin disc12Determination of S. aureus sensitivity – reviewmethod

Class 5. Oral microbiology – Bacteria causing inflammations and abscesseswithin the oral cavity; Gram-positive facultative cocci (genus Staphylococcus,Streptococcus and Enterococcus).Identification of staphylococci in selective-differential medium – mannitol salt agar (Chapman’smedium). Mannitol-positive staphylococci (S.aureus) – a change in the pigmentation of themedium from pink to yellow can be seen; mannitol-negative staphylococci (S.epidermidis) – nochange in the pigmentation13

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

Class 5. Oral microbiology – Bacteria causing inflammations and abscesseswithin the oral cavity; Gram-positive facultative cocci (genus Staphylococcus,Streptococcus and Enterococcus).The bacitracin (SP disc) test for identifying Streptococcus pyogenes16

Class 5. Oral microbiology – Bacteria causing inflammations and abscesseswithin the oral cavity; Gram-positive facultative cocci (genus Staphylococcus,Streptococcus and Enterococcus).The optochin (OP disc) test for identifying Streptococcus pneumoniae17

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

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

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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 discharge22

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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 abscesseswithin 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)24C.diphtheriae – Gram stain (Gram-positive, highlypleomorphic organisms with no particulararrangement)

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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 (withoutstorage cells)25C.pseudodiphteriticum – Gram stain (Grampositive rods, frequently show club-shapedswellings and palisade arrangement)

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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, branchingfilaments resembling the fungi)26

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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-Jensenmedium: (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 abscesseswithin 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 test28

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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 test30

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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-stainedmycobacteria)31

Class 6. Oral microbiology – Bacteria causing inflammations and abscesseswithin 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 H37Rv32

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

Class 8. Oral microbiology – Bacteriology of dental caries and periodontaldiseases.Anaerobic Gram-positive bacteria – Gram stainPropionibacterium sp. (Gram-positive bacteria, cylindrical, pleomorphic cells)34

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 .