Labquality External Quality Assessment Programmes General Bacteriology 1 4/2010
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1 Labquality External Quality Assessment Programmes General Bacteriology 1 4/2010 Photos and text: Markku Koskela, M.D., Ph.D. Clinical microbiology specialist Oulu, Finland
2 Sample 13/2010 Cerebrospinal fluid Meningitis in an adult patient Finding: Streptococcus pneumoniae ie pneumococcus as the pathogen Lab exercises: Isolation and identification of the pathogen. Antimicrobial susceptibility tests.
3 Sample 13/2010. Pneumococcus Growth on sheep blood agar and on chocolated blood agar in 5% CO 2, at +35 C after 24 hours. -> An alpha-haemolytic streptococcus
4 Sample 13/2010. Pneumococcus Gram+diplococcus Lancet shaped Test Negat control Pneumococcus: sensitivity to optochin as a diagnostic test. Bile solubility: positive
5 Sample 14/2010 Pus from leg ulcer of an old farmer Expected findings: Normal bacterial flora of the skin containing group B beta-haemolytic streptococcus (GBS) ie Streptococcus agalactiae as the pathogen. Lab exercises: Isolation and identification of the pathogens.
6 Sample 14/2010 Primary culture Growth on sheep blood agar and on chocolated blood agar in 5% CO 2, at +35 C after 24 hours. ->A slightly beta-haemolytic streptococcus and a staphylococcus.
7 Sample 14/2010 Primary culture Growth on sheep blood agar and on chocolated blood agar in 5% CO 2, at +35 C after 48 hours. ->A slightly beta-haemolytic streptococcus and a staphylococcus.
8 Sample 14/2010 GBS ie Streptococcus agalactiae Catalase: negat Gram+diplococcus or cocci in chains Pure growth on sheep blood agar plate after 24 hours. B A: neg control Streptococcal latex-test.
9 Sample 14/2010 Coagulase negative staphylococcus (Staphylococcus haemolyticus) 1 2 Staphylococcal Pure growth on sheep blood agar plate after 48 hours. Flat, glistening, white and slightly beta-haemolytic colonies resempling (?) S aureus. latex-coagulase teast: 1. Negat (S haemolyticus) 2. Posit (S aureus)
10 Sample 15/2010 Pus from frontal sinus of a 37-year-old male Findings: Fusobacterium necrophorum and Prevotella melaninogenica, as pathogens. In addition, Streptococcus viridans, representing the normal flora of the upper respiratory tract. Copathogen? Lab exercises: Isolation and identification of the pathogens.
11 Sample 15/2010 Primary aerobic culture Growth on sheep blood agar and on chocolated blood agar in 5% CO 2, at +35 C after 24 hours. -> An alpha-haemolytic streptococcus
12 Sample 15/2010. Primary anaerobic culture on anaerobic blood agar plate (FAA) 48 h, at +35 C. Inhibition of streptococcal growth around an aminoglycoside disk. Anaerobes are growing close to the disk. A large inhibition zone around a metronidazole disk caused by anaerobes. Some anaerobes are producing a brown pigment.
13 Sample 15/2010. Primary anaerobic culture Single colonies on anaerobic FAA plate. A brown, flat, glistening colony sensitive to metronidazole -> An anaerobe? A gray, flat, matt colony sensitive to metronidazole ->An anaerobe? A white colony: most probably a facultative anaerobic streptococcus.
14 Sample 15/2010. Primary anaerobic culture Additional incubation of FAA-plate up to 5 days A brown, flat and glistening colony (Prevotella sp) A gray, flat and matt colony (Fusobacterium sp)
15 Sample 15/2010. Prevotella melaninogenica 2-5 days Pigment production not as strong as on FAA plate Anaerobic growth on Kanamycin-vankomycin-laked blood-agar (KVLB)
16 Sample 15/2010. Primary anaerobic culture: EYA-plate 2 days 7 days Lipase positive colonies on Egg yolk agar (EYA). (Fusobacterium)
17 Sample 15/2010. Aerotolenrance test (48 h, +35 C) Aerobic growth on blood agar Microaerophilic growth on chocolated blood agar in 5% CO 2 Anaerobic growth on FAA-plate 1 (Fusobacterium sp) and 2 (Prevotella sp) are growing only anaerobically. Streptococcus viridans (3) is facultatively anaerobic following that it is growing on all the three plates and in all the three athmospheres.
18 Sample 15/2010. Fusobacterium necrophorum Anaerobic identification test (A I): Kanamycin S, vankomycin R, colistin S
19 Sample 15/2010. Fusobacterium necrophorum Anaerobic growth on FAA plate, 72 h. Flat, matt, haemolytic colonies. Positive lipase reaction slowly within (3) to 5 days
20 Sample 15/2010. Fusobacterium necrophorum Rapid indole test: positive Gramnegative rod
21 Sample 15/2010. Fusobacterium necrophorum - - -, - - -, - - -, + - -, - - -, RapID ANA II: Fusobacterium necrophorum (99,8 %)
22 Sample 15/2010. Fusobacterium necrophorum - - -, - - -, - - -, - - -, , - - -, - - -, - - -, Rapid ID 32 A: Fusobaterium necrophorum 71%
23 Sample 15/2010. Prevotella melaninogenica FAA 72 h: Colonies with a brown pigmentation KVLB 72 h: Slowly pigmented colonies
24 Sample 15/2010. Prevotella melaninogenica Anaerobic identification test (A I): Kanamycin S, vankomycin R, colistin S
25 Sample 15/2010. Prevotella melaninogenica Rapid indole test: negative Gramnegative rod
26 Sample 15/2010. Prevotella melaninogenica - - +, + + -, + + +, + + -, - - +, RapID ANA II: Prevotella melaninogenica (98,4 %).
27 Sample 15/2010. Prevotella melaninogenica - - +, + + +, - - -, + - -, , + - +, - - -, - + -, Rapid ID 32A: Prevotella melaninogenica (67,5 %)
28 Sample 15/2010. Streptococcus viridans An alpha-haemolytic streptococcus resistant to optochin.
29 Sample 16/2010 Amnionic fluid Finding: Listeria monocytogenes, as the pathogen Lab exercises: Isolation and identification of the pathogen.
30 Sample 16/2010 Primary aerobic culture 1 2 Catalase test: 1. Listeria monocyt. Posit 2. Negative control (Group B beta-haemolytic streptococcus, GBS) Growth on sheep blood agar plate on 5% CO 2, at +35 C, 24 to 48 h. -> Middle-sized, beta-haemolytic, convex and glistening colonies which are catalase positive.
31 Sample 16/2010: Primary aerobic culture A short grampositive rod Growth on chocolated blood agar plate in 5% CO 2 at +35 C within 24 to 48 hours. ->Middle-sized, convex and glistening colonies.
32 Sample 16/2010: Primary aerobic culture on selective Palcam agar plate Aerobic growth on a listeria-selective Palcam agar plate at +35 C within 24 to 48 hours -> Graygreen, esculin positive colonies.
33 Sample 16/2010. Listeria monocytogenes +30 C +35 C Umbrella motility at +30 C
34 Sample 16/2010. Listeria monocytogenes + - -,+ + +, + - -, - + -, + - -, - - -, + Katalaasi RapID CB Plus: Listeria monocytogenes (42%) Listeria innocua (37%) The strain is beta-haemolytic -> Listeria monocytogenes
35 Sample 16/2010. Listeria monocytogenes - + +, + - +, + - -, - API Listeria: 6510 Listeria monocytogenes ( 99 %)
36 The first snow in November
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