Labquality External Quality Assessment Programmes General Bacteriology 1 2/2014
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1 Labquality External Quality Assessment Programmes General Bacteriology 1 2/2014 Photos and text: Markku Koskela, M.D., Ph.D. Clinical microbiology specialist NordLab Oulu, Finland
2 Specimen 21/2014 Pus from infected stab wound in the leg Finding: Streptococcus pyogenes, significant pathogen. Lab exercises: Isolation and identification of the pathogen. Antimicrobial susceptibility tests.
3 Specimen 21/2014. Pus from stab wound Growth on sheep blood agar plate in 5% CO 2 at 35 C after 24 to 48 hours. -> Middle-sized, beta-haemolytic streptococcal colonies
4 Specimen 21/2014. Pus from stab wound Growth on chocolated blood agar plate in 5% CO 2 at 35 C after 24 to 48 hours. -> Small, gray coccal colonies.
5 Specimen 21/2014 (1/2012). Streptococcus pyogenes Gram-positive cocci in chains A + G - Growth on sheep blood agar plate in 5% CO 2 at 35 C after 24 to 48 hours. -> Middle-sized, gray-white coccal colonies surrounded by a large beta-haemolytic zone. Streptococcal latex agglutination test
6 Specimen 22/2014 Excretion from itching outer auditory canal of a 72-year-old male. Findings: Proteus mirabilis, significant pathogen. Acinetobacter lwoffii, normal flora of outer auditory canal. Lab exercises: Isolation and identification of the bacteria. Interpretation of the findings.
7 Specimen 22/2014 Excretion from itching outer auditory canal of a 72-year-old male. Growth on sheep blood agar plate in 5% CO 2 at 35 C after 24 to 48 hours. 1. Large, gray, convex and glistening bacillar colonies which are spreading all over the plate. (Smell of Proteus sp.) 2. Small, gray, convex and glistening bacillar colonies.
8 Specimen 22/2014 Excretion from itching outer auditory canal Growth on chocolated blood agar plate in 5% CO 2 at 35 C after 24 to 48 hours. 1. Large, gray, convex and spreading bacillar colonies. 2. Small, gray, convex bacillar colonies.
9 Specimen 22/2014 Excretion from itching outer auditory canal Growth on Cled-agar plate, at 35 C, after 24 to 48 hours. 1. Middle-sized, pale green, flat, lactose-negative colonies. 2. Small, green, convex, lactose-negative colonies.
10 Specimen 22/2014 Excretion from itching outer auditory canal Growth on BBL Chromagar Orientation plate, at 35 C,after 24 to 48 hours. 1. Middle-sized, flat, brownyellow colonies. 2. Small, convex, glistening brown-yellow colonies.
11 Specimen 22/2014 (2/2009). 1. Proteus mirabilis Growth on sheep blood agar plate in 5% CO 2 at 35 C after 24 hours. Gram-negative rod 1. Large, gray, convex and glistening bacillar colonies which are rapidly spreading all over the plate. (Smell of Proteus sp.)
12 Specimen 22/ Proteus mirabilis Fermentation test results: 1) Glucose/gas +/- (yellow/no gas) 2) Indole - (colourless) 3) motile 4) citrate + (blue) 5) urea + (red) 6) TSI: red/yellow/gas on bottom, H2S positive (black). A fermentative bacterium. ->API20E
13 Specimen 22/ Proteus mirabilis 1. API20E Proteus mirabilis (99,9%). Oxidase - - -, + + +, + + -, - + +, - - -, - - -, Vitek V2 GN Proteus mirabilis
14 Specimen 22/ Acinetobacter lwoffii Growth on sheep blood agar plate in 5% CO 2 at 35 C after 24 hours. Small, convex and glistening colonies. Gram-negative coccobacillus
15 Specimen 22/ Acinetobacter lwoffii Fermentation test results: 1) Glucose/gas -/- (green/no gas) 2) Indole - (colourless) 3) non-motile 4) citrate - (green) 5) urea - (yellow) 6) TSI: red/red/no gas, H2S negative. A fermentative or a non-fermentative bacterium? ->API20E/NE?
16 Specimen 22/ Acinetobacter lwoffii 1. API20E No result 2. API20NE Acinetobacter lwoffii (99%) (After incubation for 48 hours!). 3. Vitek-2 GN No result
17 Specimen 23/2014 Pus from breast abscess of a 47-year-old female Findings: Prevotella intermedia/nicrescens and Prevotella oralis, significant pathogens. Lab exercises: Isolation and identification of the bacteria. Interpretation of the findings.
18 Specimen 23/2014 Pus from breast abscess Gram-stain of the specimen: Gram-negative rods
19 Specimen 23/2014 Pus from breast abscess. Primary aerobic culture Cultivation on sheep bood and chocolated blood agar plates. Aerobic incubation in 5% CO 2 at 35 C for 7 days. No growth.
20 Specimen 23/2014 Pus from breast abscess. Primary anaerobic culture Growth on anaerobic blood agar platela (FAA) 48 hours at 35 C. Inhibition zone around metronidazole disk ->anaerobes!
21 Specimen 23/2014 Pus from breast abscess. Primary anaerobic culture Growth on anaerobic blood agar platela (FAA) 48 hours at 35 C. Small and very small colonies ->Aerotolerance tests and Gram-stain!
22 Specimen 23/2014 Pus from breast abscess. Primary anaerobic culture Anaerobic culture on KVLB-plate at 35 C for 48 hours. 1. Dark, strongly pigmented colonies. 2. Very small, light colonies. ->Aerotolerance test and Gram-stain.
23 Specimen 23/2014 Pus from breast abscess. Primary anaerobic culture Anaerobic culture on KVLB-plate at 35 C for 4 days. 1. Dark, strongly pigmented colonies turning to yellow. 2. Very small, light yellow colonies.
24 Specimen 23/2014 Aerotolerance tests Blood agar: aerobic; chocolated blood agar: 5% CO 2, FAA: anaerobic 1. Left: The bacterium is growing only on the anaerobic FAA-plate. It is an obligative anaerobe. (Prevotella intermedia). 2. Rigth: The bacterium is growing only on the anaerobic FAA-plate. It is an obligative anaerobe. (Prevotella oralis).
25 Specimen 23/2014. (Anaerobe course 2011). 1. Prevotella intermedia/nicrescens FAA KVLB Anaerobic growth on FAA- and KVLB-plates at 35 C after 2 to 3 days. Small to middle-size, brown-pigmented, glistening, convex colonies. The pigmentation is stronger on a KVLB-plate.
26 Specimen 23/2014. (Anaerobe course 2011). 1. Prevotella intermedia/nicrescens EYA after 4 days Catalase negative (No bubling) Spot indole test positive (blue) Growth on EYA-plate after 4 days at 35 C. Lipase-reaction positive: oily colonies. Lesithinase-reaction negative: The colonies are not surrounded by a white precipitate.
27 Specimen 23/2014. (Anaerobe course 2011). 1. Prevotella intermedia/nicrescens Anaerobe identification test (A I): Kanamycin R, vankomycin R, colistin S. Metronidazole S. A Gram-negative anaerobic bacillus. Gram-negative rod
28 Specimen 23/2014 (Anaerobe course 2011). 1. Prevotella intermedia/nicrescens 1. RapID ANA II Prevotella intermedia (100%) - - -, - + -, - + -, + + -, - - +, - - +
29 Specimen 23/ Prevotella intermedia/nicrescens 1. Rapid ID 32A Prevotella intermedia (99%)
30 Specimen 23/ Prevotella oralis Catalase negat. (No bubling) Anaerobic growth on FAA-plate at 35 C, after 3 days. Convex, glistening, yellowish colonies. Spot indole test negative (colourless)
31 Specimen 23/ Prevotella oralis Anaerobe identification test (A I): Gram-negative rod Kanamycin R, vankomycin R, colistin S. Metronidazole S. A Gram-negative anaerobic bacillus.
32 Specimen 23/ Prevotella oralis 1. RapID ANA II Incorrect result, but if Prevotella oralis 100% - - -, + + -, + + +, + + +, - - -, - - -
33 Specimen 23/ Prevotella oralis 1. Rapid ID32A Prevotella oralis 73%
34 Specimen 24/2014. A sample taken during surgery operation from ureter of a 57-year-old female. Finding: Oligella ureolytica, significant pathogen. Lab exercises: Isolation and identification of the bacterium. Interpretation of the finding.
35 Specimen 24/2014. A pus sample taken during surgery operation from ureter of a 57-year-old female. Gram-stain of the specimen: Gram-negative rods
36 Specimen 24/2014. Pus from ureter. Primary aerobic culture Growth on sheep blood agar plate in 5% CO 2 at 35 C after 24 to 48 hours. -> Small, dry, convex colonies, which are oxidase positive.
37 Specimen 24/2014. Pus from ureter. Primary aerobic culture Growth on chocolated blood agar plate in 5% CO 2 at 35 C after 24 to 48 hours. -> Small, dry, convex colonies, which are catalase positive.
38 Specimen 24/2014. Pus from ureter. Primary aerobic culture No growth on Cled-agar at 35 C after 48 hours.
39 Specimen 24/2014. Pus from ureter. Primary aerobic culture No growth on BBL Chromagar Orientation-plate at 35 C after 48 hours.
40 Specimen 24/2014. Pus from ureter. Primary anaerobic culture No growth on anaerobic blood agar plate after 7 days.
41 Specimen 24/2014. Oligella ureolytica Oxidase + Aerobic growth on sheep blood agar at 35 C after 48 hours. Small, convex colonies. Gram-negative rod
42 Specimen 23/ Oligella ureolytica 1. RapID NH 2001 Oligella ureolytica (99%) - + -, - - -, - - -, API20NE Oligella ureolytica (97%) 3. Vitek-2 No result
43 Have a Nice Summertime!
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