Labquality External Quality Assesment Programmes General Bacteriology 1 3/2010

Transcription

1 Labquality External Quality Assesment Programmes General Bacteriology 1 3/2010 Photos and text: Markku Koskela, M.D., Ph.D. Clinical microbiology specialist Oulu, Finland

2 Patient and sample 9/2010 Pus from a skin pustule. Pemphigus in a neonate. Finding: Staphylococcus aureus, pathogen Lab exercises: Isolation and identification of the pathogen. Antimicrobial susceptibility tests.

3 Sample 9/2010. Staphylococcus aureus Growth on sheep blood and chocolated blood agar plates in ambient air with 5% CO 2, at +35 C, after 24 hours. Medium sized, flat, glistening, yellowish, beta-haemolytic colonies -> S.aureus?

4 Sample 9/2010. S.aureus Gram positive cocci in clusters Staphylococcal Latex-coagulase test: positive Negative control

5 Patient and sample 10/2010 Middle ear effusion from a three years old child with recurring acute otitis media Findings: Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus, all three as pathogens Lab exercises: Isolation and identification of the pathogens.

6 Sample 10/2010 Middle ear effusion Growth on a sheep blood agar plate in ambient air with 5% CO 2, at +35 C, within 24 to 48 hours. 1. Flat, glistening, yellowish, beta-haemolytic S.aureuscolonies 2. with Haemophilus sp. as satellites 3. Small, dome-shaped colonies that can be moved on the agar surface -> oxidase + ->Moraxella catarrhalis?

7 Sample 10/2010 Middle ear effusion Growth on a chocolated blood agar plate in ambient air with 5% CO 2, at +35 C, within 24 to 48 hours 2. Dome-shaped, graywhite M.catarrhalis-colonies 3. Great, flat, yellowish S.aureus-colonies 1. Gray, flat Haemophiluscolonies

8 Sample 10/2010 Haemophilus influenzae S aureus Satellite test+ Growth on a chocolated blood agar Small pleomorphic Gram-negative rods Growth factors: X and V +

9 Sample 10/2010 Moraxella catarrhalis Gram-negat diplococci Oxidase+ Tributyrene test +

10 Sample 10/2010. Staphylococcus aureus Coagulase test + Compare with the sample 9/2010

11 Sample 11/2010 Pus from an ovarial abscess of a 43-years old female Findings: Klebsiella pneumoniae and Actinomyces israelii as pathogens. Lab exercises: Isolation and identification of the pathogens. How to isolated fastidious and slowly growing anaerobes from among rapidly and heavily growing aerobic bacteria!?

12 Sample 11/2010 Pus from an ovarial abscess Gram stain of the (simulated) native pus sample: Gram negative rods and thin, branching Gram(?) rods!

13 Sample 11/2010 Pus from an ovarial abscess Growth on a sheep blood agar plate in ambient air with 5% CO 2, at +35 C, within 24 to 48 hours. Aerobic slime forming bacillar colonies. Something else?

14 Sample 11/2010 Pus from an ovarial abscess Gowth on a chocolated agar plate in ambient air with 5% CO 2,, at +35 C, within 24 to 48 hours Aerobic bacillar colonies. More?

15 Sample 11/2010 Pus from an ovarial abscess Growth on an anaerobic blood agar (FAA) plate after anaerobic incubation for 2 and 7 days at +35 C. 2 days 7 days Klebsiella colonies. No signs of anaerobes. Dry, white, molar-like colonies among the Klebsiella colonies.

16 Sample 11/2010 Pus from an ovarial abscess Growth on another anaerobic blood agar (FAA) plate after anaerobic incubation for 2 and 7 days at +35 C. White, molar-like colonies ->Actinomyces sp? See more detail with 200 to 400% magnification!

17 Sample 11/2010 Pus from an ovarial abscess Growth on an anaerobic phenylethylalcohol (PEA) agar plate after anaerobic incubation for 7 days at +35 C. Only Gram-positive bacteria are growing well on PEA agar. Actinomyces colonies

18 Sample 11/2010 K.pneumoniae Fermentation tests findings of Klebsiella pneumoniae: 1) Glucose +/+ (yellow /gas production positive) 2) Indole (colorless) 3) nonmotile 4) citrate + (blue) 5) urea + (red), TSI (yellow/ yellow/gas+, H2S production negative)

19 Sample 1/2010. K.pneumoniae - - +, - + -,+ - -, + - +, + + +, + + +, Oxid API20E Klebsiella pneumoniae (98%).

20 Sample 4/2010. Actinomyces israelii Aerotolerans test (24 to 48 hrs at +35 C) 1. Aerobic blood agar: No growth 2. Chocolated blood agar: No growth 3. Anaerobic FAA-agar: Positive growth +++.

21 Sample 4/2010. Actinomyces israelii Preliminary identification of anaerobes with antibiotic disks: Kanamycin S, Vankomycin S and Colistin R ->A Gram-positive anaerobe Metronidazole R Typical for Actinomyces sp. -like Gram-positive anaerobes.

22 Sample 4/2010. Actinomyces israelii White molars on FAA plate after 7 days anaerobic incubation at +35 C. Gram-positive branching rods Catalase negative

23 Sample 11/2010. Actinomyces israelii - + +, + + +, + - -, - + +, + + +, RapID ANA II Actinomyces israelii (99,7 %)

24 Sample 11/2010. Actinomyces israelii - + +, + - +, + + -, - + +, , - + -, + + -, + + -, Rapid ID32 A Actinomyces israelii (84 %)

25 Sample 12/2010 Sputum sample of a farmer with Hodgkin disease Findings: Rhodococcus equi, pathogen. Alpha-haemolytic streptocci as normal flora of the upper respiratory tract. Lab exercises: Isolation and identification of the pathogen.

26 Sample 12/2010: Sputum Growth on a sheep blood and chocolated blood agar plates in ambient air with 5% CO2, at +35 C, within 24 hours. -> 1) Glistening, slimy, irregular colonies 2) alpha-haemolytic streptococcus colonies

27 Sample 12/2010: Sputum Growth on a sheep blood and chocolated blood agar plates in ambient air with 5% CO2, at +35 C, within 48 hours. -> 1) Glistening, slimy, irregular colonies 2) and some alpha-haemolytic streptococcus colonies

28 Sample 12/2010. Rhodococcus equi Gram-positive coccoid bacillae Catalase+ A salmon-colored pigment of the colonies appeared on a blood agar plate just after aerobic incubation for 5 to 7 days

29 Sample 12/2010. Rhodococcus equi - - -, - + +, - - -, + + +, + - +, RapID CB Plus: Rhodococcus equi ( 99,9 %)

30 Sample 12/2010. Alpha-haemolytic streptococcus An optocin-resistent alpha-haemolytic streptococcus of the normal flora of the upper respiratory tract

31 Autumnal greetings from a Finnish forest covered with Reindeer lichen (Cladolia rangiferina) among which

32 also edible boletus (Boletus edulis) is growing.