Laboratory Training and Procedures Bacteriological Techniques Sputum smear Antoine Pierson (BiolTrop)

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1 Laboratory Training and Procedures Bacteriological Techniques Sputum smear Antoine Pierson (BiolTrop) Cocci gram positif Bacterial species Macroscopique Culture media Microscopic appearance Reference characteristics aspect Routine Oxidase Catalase Respiration Staphylococcu Aureus Creamy yellow colonies, ± hemolytic, sometimes white, Chocolate agar, Chapman DE : small cocci in clusters or isolated - + Aero-anaerobic Negative coagulase Creamy white colonies, slightly or not hemolytic, Chocolate agar, Chapman DE: cocci in clusters or isolated ± large - + Aero-anaerobic Micrococcus White colonies, fine, sometimes dry, Chocolate agar, Chapman DE: large cocci in clusters or tetrads + (-) + Strictly aerobic Stomatococcus Viscous, elastic colonies (look like chewing-gum), Chocolate agar, Chapman DE: large cocci in clusters - + / - Aero-anaerobic Streptococaceae Very fine grey colonies Alpha hemolysis (green) Cocci in chains, sometimes very long - + Aero-anaerob Strep. Alpha-hemolytic Very fine (Alpha) or grey Cocci in chains,

2 Strep. Beta-hemolytic (Beta) colonies sometimes long Beta hemolysis Streptococaceae Pneumoniae Fine colonies, sometimes mucoid colonies, referred to as dew drop-like, or slightly pitted at the center Alpha hemolysis Cocci in chains, Diplococci (candle flame) Short chains Occasionally agglutinated on rich culture (lysis) Enterococcus Grey medium-sized colonies, sometimes hemolytic BCP (urine) Cocci in short chains, slightly elongated Streptococcaceae Aerococcus Fine white colonies, Alpha hemolysis Cocci in clusters or tetrads - - Micro-aerophilic Leuconostoc Fine whitish colonies, nonhemolytic Coccobac. cocci, sometimes in short chains, diplococci or clusters Pediococcus Idem Large cocci in clusters, pairs or tetrads - - Micro-aerophilic Peptostreptococcacea Peptostreptococcus or Fine colonies Cocci in chains, diplococci - - Anaerobic Peptococcus

3 Gram negative cocci Bacterial species Macroscopique Culture media Microscopic appearance Reference characteristics aspect Routine Oxidase Catalase Respiration Neisseriaceae Neisseria Grey mucoid mediumsized colonies Diplococci (coffee beans) + + Strictly aerobic Meningitidis Gonorrhae Grey small-sized colonies Idem + + Strictly aerobic Saprophytes (mucosa, perflava) Rough, yellow or white small-sized colonies Idem + + Strictly aerobic Diplococci Brahamella catarrhalis White medium-sized colonies of dryish aspect (coffee beans), sometimes slightly + + Strictly aerobic elongated Veillonellaceae White anaerobic colonies Diplococci + + Anaerobic Veillonella

4 Gram positive bacilli Bacterial species Macroscopique Culture media Microscopic appearance Reference characteristics aspect Routine Oxidase Catalase Respiration Corynebacteriaceae Corynebacterium Fine grey or white colonies Tapered rocket-shaped bacilli, of varying length, arranged in vertical rows - + Aero-anaerobic Arcanobacterium Fine colonies Coccobacilli - Strictly aerobic Arcanobacterium Small colonies sometimes beta hemolytic Small bacilli - Aero-anaerobic Rhodococcus Small colonies Tapered bacilli + Aero-anaerobic Oerskovia Brilliant colonies pigmented with yellow Mobile bacilli + Aero-anaerobic Bcillaceae Bacillus Large mucoid colonies, sometimes hemolytic Large bacilli, sometimes spore-forming, mobile, sometimes Gram - + / - + Aero-anaerobic Lactobacillaceae Lactobacillus Fine colonies, sometimes hemolytic Long fine bacilli, also maybe polymorphous depending on species Medium-sized colonies, Small bacilli with specific - + Aero-anaerobic

5 AeG+ incertaesedis beta hemolytic, similar to mobility, arranged in strep colonies vertical rows Listeria Erysipelothrix Very fine colonies Fine bacilli, sometimes granular and filamentous Clostridiaceae Mucoid colonies, sometimes iridescent, of Large bacilli with square ends, sometimes spore- Clostridium varying size forming ANAG+ non sporulé Actinomyces Creamy white colonies, very slow to grow (5-10 days) Small straight or curved bacilli with ramifications or "branched" bacilli - + Anaerobic (facultative) Bifidobacterium Fine colonies Small bacilli with 2 branches, polymorphous - - Anaerobic Eubacterium Fine colonies Small coccobacilli or bacilli, depending on the species - - Strictly anaerobic Propionibacterium Fine colonies, grow in 2-4 days Rocket-shaped bacilli (see Corynebacteria) - + Strictly anaerobic Mobiluncus Fine colonies, grow in 4-5 days Incurved, very mobile bacilli - - Strictly anaerobic Actinomycetales White colonies Sabouraud agar Filamentous, granulous alcohol resistant bacilli - + Aerobic Nocardia

6 Gram negative bacilli Bacterial species Macroscopique Culture media Microscopic appearance Reference characteristics aspect Routine Oxidase Catalase Respiration Enterobactériaceae (Escherichiae, Salmonella, Shigella, Proteus, Klebsiella, Enterobacter...) Large colonies, often mucoid colonies Hektoën agar Polymorphous bacilli, mobile (unless exceptional) - + Aero-anaerobic Moraxellaceae Acinetobacter Large mucoid colonies Coccobacilli, often isolated Capsulated - + Strictly aerobic Moraxella Fine colonies Blood aga Coccobacilli + + Strictly aerobic Pseudomonaceae (Pseudomonas, Xanthomonas, Alcaligenes, Flavobacterium) Mucoid colonies of varing size, brilliant Columbia agar Very mobile fine bacilli + unless excep. + Strictly aerobic Vibrionaceae (Vibrion, Aeromonas, Large flat smooth colonies Very mobile bacilli + + Strictly aerobic Plesiomonas) Haemophilaceae Haemophilus Fine grey colonies (mucoid or dry) Long fine bacilli, sometimes entangled + (slow) + Aero-anaerobic Brucella Small, grey colonies (slow to culture - 3 weeks) Very small coccobacilli +- + Aero-anaerobic

7 Pasteurellaceae Pasteurella Grey mucoid colonies of varying sizes Small cocci-shaped bacilli, more or less isolated + - Strictly aerobic Francisella Difficult to grow Small bacilli - + Strictly aerobic AeG-incertaesedis Gardnerella Fine hemolytic colonies Human blood agar Small Gram- coccishaped bacilli - (=Haemophilus), but also Gram + bacilli (Corynebacteria) - - Aero-anaerobic prefers anaerobic Eikenella Fine dry colonies encrusting in the agar, slow to culture (2 to 3 days) Small bacilli + - Fines grey colonies, slow Kingella and difficult to culture. One species is Beta hemolytic Columbia/blood agar Coccobacilli, sometimes in pairs or short chains + + Bacteroïdaceae: Bacteroïde Medium-sized mucoid colonies sometimes pigmented Small bacilli + / - Strictly anaerobic Prevotella Small, grey, brilliant colonies, sometimes pigmented, slow to grow Very small coccobacilli - - Capnocytophaga Small colonies, slow to grow (2-3 days) Small bacilli + + / - Microaerophilic Mucoid medium-sized Elongated rocket- - - Strictly anaerobic

8 Fusobacteriaceae Fusobacterium colonies, strong odour shaped bacilli Campylobacteraceae Genre : Campylobacte Grey, medium-sized colonies Campylo selective medium Curved, spiral, mobile bacilli + + Micro-aerophilic Helicobacter Grey mucoid colonies, slow to grow (3-4 days) in microaerophilic medium Curved, spiral seagull wing-shaped bacilli + + Micro-aerophilic