E-visitation of practicals A small guide to practical examination by Zahradníček This is the survey of the most imporant things. Nevertheless, you can be asked everything from practical sessions of both Microbiology I and Microbiology II. Microscopy v Gram staining o to be able to perform it o to be able to observe a preparation and to identify G+/G- cocci/bacilli (+arrangement), yeasts, epithelial cells, WBCs o to know the principle v Wet mount, other staining methods perfomed in practicals (survey) v (Ziehl-Neelsen staining, see Acid fast bacteria) v Interpretation of microscopical findings (importance of epithelial cells, leucocytes) Culture v Most important culture media o to be able to recognize blood agar, Endo agar and Mueller Hinton agar o to be able to describe function of all fourteen media from J03 v Inoculation (to be able to inoculate a strain/a swab) v Description of colonies (practically) Biochemical identification v Catalase test o to be able to do it o to understand its practicle o to be able to give an example of its use in diagnostics v Strip tests o to know the most important ones (oxidase, PYR, INAC) and to give examples of using them o to be able to use them practically (incl. reading the results) v Hajna, MIU and similar tests o to know their practical use and what do they detect v Enterotest-like tests o to be able to read an Entero- (Staphy- ...) –test and to describe its principle Outer influences, disinfection and sterilisation v To know the safety rules in the laboratory v To know the most common disinfectants and sterilization methods and the way they are used (chloramin, NaOCl, Ca(OCl)[2], iodine-povidone, hydrogen peroxide, peracetic acid, ajatin, UV-rays disinfection, hot air sterilisation, steam sterilization, radiation sterilization) v To understand the methodological difference between testing of growth limit and survival limit v To be able to read corresponding tests (see Task 1 from P06) v To know how effect of disinfection and sterilization can be tested Antimicrobial drugs v To know principles of microdilution test, diffusion disk test and E-test, to be able to read the results of all of them and to interprete them v To understant the importance of MIC and its comparison with breakpoint level v To know basic methods of testing factors of resistance (betalactamases) Serological tests (J07 to J10) v To be able to read the results any of these tests; students will get the necessary information (dillution in the first well, c. o. counting in ELISA etc.) v To be able to describe the basic indication for the test and to interprete these results in combination with other parameters; including ASO! v To understand principle of antigen/analysis reactions and its use for antigen detection in a specimen/antigen analysis of a strain/antibody detection v To understand major interpretation difference between direct and indirect diagnostical methods v To know principles of agglutination, precipitation, agglutination on carriers, CFT, neutralisation (ASO, HIT, VNT), reactions with labelled components, western blotting, incl. differences between them v To understand titers, titer dynamics, seroconversion, importance of IgM/IgG (and knowing what reactions enable their detection – importance of conjugate), avidity (A-wishing students) v To be able to construct a scheme of HBsAg and anti-HBs testing v To understand terms "heterophilic antibodies" and "anticomplementarity test" Detection of nucleic acid v To know the basic indication for these methods in microbiology v To understand the difference between methods with/without amplification v To know basic principle of the reaction, including two major ways of product detection v To understand the imporance of internal control v To be able to read practically a PCR result (on a picture), including IC result interpretation Virology v To know the ways of isolation of a virus (including individual structures of a fertilized egg) v To be able to differenciate a cell culture with/without CPE (in easy cases only) and to understand, what a CPE is v (plus serology: HIT, VNT, see serology) Easily culturable bacteria and yeasts (P01–P06; P10) v To be able to find out (and utilize practically) a diagnostic algorithm to identify common bacteria except G+ rods (Staphylococcus aureus, coagulase-negative staphylococci, Streptococcus pyogenes, S. agalactiae, S. non-A-non-B, S. pneumoniae, oral streptococci, Enterococcus faecalis, E. faecium, Escherichia coli, Klebsiella pneumoniae, Salmonella enterica, Proteus sp., Pseudomonas aeruginosa, other G- non-fermenters, Haemophilus influenzae, H. parainfluenzae, Pasteurella multocida, Neisseria gonorrhoeae, Neisseria meningitidis, oral neisseriae, Moraxella catarrhalis, Candida albicans, Candida sp.) v For G+ rods: to know their main characteristics; to be able to identify practically coryneform rods according to their pallisade arrangement Anaerobic bacteria v To be able to describe an anaerobic jar and an anaerobic box, their parts and their function v For clostridia: to know their main characteristics; to be able to identify C. tetani according to its sphaerical terminal endospore Acid-fast rods v To know the principle of Ziehl-Neelsen staining, to be able to distinguish between pictures of positive and negative findings and pictures stained using other staining methods v To know the principles of acid-fast rod culture, to know basic media, to be able to distinguish pictures of positive findings/negative findings/pictures describing something other Spiral bacteria v To explain use (and complications in use) of direct methods in spirochete diagnostics v To understand screening/confirmatory reactions for Borrelia and Treponema v To be able to read and interprete the tests (see also Serology) Fungi v To know basic diagnostic methods used in mycology v To be able to read a microprecipitation test for lung aspergillosis and to explain its principle v To know the basic principles of sampling for mycology v See also „Easily culturable bacteria and yeasts (P01–P06; P10)“ Parasites v To know basic methods for parasites (Faust, Kato, Graham; thick and thin smear; C. A. T. swab and Giemsa stained smear for trichomonas; indirect diagnostics of tissue parasites) v To be able to distinguish the most common helmint eggs (tapeworm, pinworm, common roundworm, Trichuris) and tapeworm proglottid v To know the basic principles of sampling for parazitology Biofilm v To know the diagnostic methods of biofilm detection v To know the difference bethween three most typical methods of venous catether microbiological diagnostic v To be able to read the results of a biofilm : glucose/time experiment (see P12 Task 4) v To be able to read MBEC values and to interprete the result (in comparison with MIC) Clinical microbiology v To be able to find a pathogen in phagyngeal flora (and to know the composition of normal pharyngeal flora, and common pharyngeal pathogens) v To be able to read a result of urine culure semiquantitativelly and qualitativelly v For a simple mini-casuistics, to be able to find out the best sampling method, including finding the best swab or container (practically) v To understand basic principles of sampling under various circumstances The taks could be rather practical (e. g. „Gram stain a given strain“, „Read the results of an Enterosest“, „Read and interprete results of tests for syphilis“) or more theoretical („Among three given strains, find a Staphylococcus and determine more precisely“ – here majority of steps would be done only „orally“). 5th December 2008 Ondřej Zahradníček