1. MICROBIOLOGY LABWORK JPEMS 2012 NANTES Dr C. Bressollette (Virology, Nantes) Dr J. Caillon (Bacteriology, Nantes) Dr M. Eveillard (Bacteriology, Angers) Cécile Peltier (Virology, Nantes)

2. Safety rules  Wash your hands at the beginning and the end of the session  Wear a labcoat  Wear disposables gloves when recommended (IHA, PCR)  For some of the bacteriology procedures, you will work around the flame of a Bunsen burner > long hair must be restrained  Throw the « infected material » in the biohazard container  Clean small spills with bleach  Disinfect work areas after use

3.  Virology –Viral cultures: Observation of a cytopathic effect HSV typing by immunofluorescence –Molecular assay Amplification of a viral gene –Serologic assay: Inhibition of hemagglutination  Bacteriology  Analysis of a bacterial mixture  Gram staining  Bacterial identification  Susceptibility to antibiotics  Analysis of the environment The week schedule

4. Day 1 - Virology The laminar flow hood / biosafety hood Plastic culture flasks

5. Day 1 - Virology  Observation of non infected adherent cells under the microscope Human fibroblasts Primary cells, limited multiplication in vitro HEP-2: epithelial cells, immortalized cell line

6. Day 1 - Bacteriology Study of a mix of two bacterial strains  6 different bacterial mixtures: A, B, C, D, E : mark the letter on your mix tube  Microscopic observation –Smear colored by the method of gram staining See the demonstration and technique Pink bacteria: gram negative Blue bacteria: gram positive  Bacterial isolation on an agar medium (quadrant method) See the demonstration and your workbook 3 plates –Chromogenic media –Drigalski plate (green) – Chapman plate (pink):