Practical Microbiology

based on the Hungarian practical notes entitled "Mikrobiológiai Laboratóriumi Gyakorlatok"

Erika M. Tóth, Andrea K. Borsodi, Tamás Felföldi, Balázs Vajna, Rita Sipos and Károly Márialigeti

Authors of the Hungarian and English versions 

Csaba Romsics, Judit Makk, Katalin Jáger, Márton Palatinszky and Éva Ács

Authors of the Hungarian version 

Erika M. Tóth and Károly Márialigeti

Editors of the English version 

Attila Náfrádi

Language assistant 

Table of Contents

1. PREFACE
2. WORK IN A MICROBIOLOGICAL LABORATORY
2.1. Biological safety level categories and the airborne route of pathogen transmission
2.2. The principle of containment, the setup and basic pieces of equipment of a microbiological laboratory
2.3. Biological safety cabinets
2.4. Behaviour and work in a microbiological laboratory
2.4 1. What to do in case of biological spill (involving BSL2 microorganisms)
2.4.2. Decontaminating hygienic hand wash and personal decontamination
2.4.3. Emergency and first aid guide
3. STERILISATION AND DISINFECTION
3.1. Procedures of sterilisation
3.1.1. Sterilisation by heat
3.1.2. Sterilisation by radiation
3.1.3. Filter sterilisation
3.1.4. Sterilisation by chemicals
3.2. Procedures of disinfection
3.3. Control of the efficacy of sterilisation equipment
3.4. Determination of the microbiological efficacy of disinfectants
4. SAMPLING METHODS IN MICROBIOLOGY
4.1. Sampling for diagnostic purposes
4.2. Sampling from various environments
4.2.1. Collection of air samples
4.2.2. Collection of soil samples
4.2.3. Collection of water samples
4.2.4. Sampling the surface of objects
4.2.5. Hygienic control of the hands of operators
5. INTRODUCTION TO THE USE OF PRACTICAL LABORATORY MICROSCOPES
5.1. Bright-field light microscopy
5.2. Fluorescence microscopy
6. CELL- AND GERM-COUNTING METHODS
6.1. Determination of cell counts with microscope
6.2. PCR-based cell counts
6.3. Determination of germ counts based on cultivation
6.3.1. CFU-counting techniques
6.3.2. The membrane filter technique
6.3.3. The end point dilution technique (MPN method)
7. STRAIN CULTURE AND CULTIVATION-BASED TECHNIQUES
7.1. Demonstration of microbes in the environment
7.2. Preparation of microbiological culture media
7.3. Basic bacterial cultivation techniques
7.3.1. Enrichment
7.3.2. Spread plate and pour plate methods
7.3.3. Isolation and streak plate technique
7.3.4. Anaerobic cultivation techniques
7.3.5. Transfer, maintenance and storage of pure cultures
7.4. Pheno- and genotypic characterisation of bacterial strains
7.4.1. Colony- and cell morphology, staining procedures
7.4.2. Study of bacterial enzymes
7.4.3. Physiological and ecological studies on bacterial strains
7.4.4. Study of the effect of antimicrobial compounds and antibiotics
7.4.5. Chemotaxonomical studies of bacterial strains
7.4.6. Bacterial species identification based on 16S rDNA sequence homology
8. ANALYSIS OF THE MICROBIOLOGICAL QUALITY OF DIFFERENT ENVIRONMENTS
8.1. Microbiology of surface waters and wastewaters, hygienic control
8.2. Soil examinations
8.3. Examination of microorganisms participating in the nitrogen cycle
8.4. Examination of microorganisms participating in the sulphur cycle
9. FERMENTATION PROCESSES IN BIOTECHNOLOGY
10. DATA ANALYSIS – TAXOMETRICS
11. BASIC ALGOLOGICAL METHODS
12. SUPPLEMENTARY MATERIAL
12.1 Supplementary exercises
12.2. Taxon information sheets with supplementary figures
12.3. Test results with supplementary figures
13. LIST OF EXERCISES
14. APPENDIX
14.1. Using LABOVAL 4 type microscopes for bright-field light microscopic observations
14.2. Culture media used in the practical
14.3. Dyes, reagents and solutions used in the practical
14.4. Tables
15. WORKING DEFINITIONS
16. REVIEW QUESTIONS
17. REFERENCES