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AGRICULTURAL AND ENVIRONMENTAL MICROBIOLOGY Department of Microbiology , Nutrition and Dietetics. Course syllabus – laboratory exercises Lecturer: Prof. Vojtěch Rada Office No.: 221 rada @ af.czu.cz. Course syllabus – laboratory exercises.

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agricultural and environmental microbiology department of microbiology nutrition and dietetics
AGRICULTURAL AND ENVIRONMENTAL MICROBIOLOGYDepartment of Microbiology, Nutrition and Dietetics
  • Course syllabus – laboratory exercises
  • Lecturer: Prof. Vojtěch Rada
  • Office No.: 221
  • rada@af.czu.cz
course syllabus laboratory exercises
Course syllabus – laboratory exercises
  • Microscopyofbacteria: Negative staining, simplestaining
  • Microscopyofbacteria: Gram staining
  • Yeast study: Methylenebluestaining
  • Mould study: Zygomycetes (Mucor, Rhizopus), Ascomycetes (Eupenicillium) and Deuteromycetes (Penicillium, Aspergillus. Asexual reproduction (conidiospores and sporangiospores)
  • Actinomycetesandantibiotics
course syllabus laboratory exercises1
Course syllabus – laboratory exercises
  • Identification of bacteria (staphylococci)
  • Microbiology of drinking water
  • Microbiology of milk: fermented milk products, starter cultures.
  • Carbon cycle: amylolytic bacteria
  • Nitrogen cycle: Nitrogen fixing bacteria (Azotobacter, Rhizobium)
laboratory safety
LABORATORY SAFETY
  • Do not drink, eat and smoke
  • Protective clothing
  • Aseptic technique
  • Bacteriological loop, needle
  • Bunsen burner
  • Bacteriological stains
brightfield microscopy
Brightfield microscopy
  • low-power objectives (4-20x)
  • high-dry objectives (40-60x)
  • immersion objectives (90-100x)
  • Resolution limit (0.4 μm)
  • Magnification(1500x)
  • Oil immersion technique.
slide6

PROTOZA 100 μm

MOULDS AND YEASTS 5 – 10 μm

BACTERIA - COCCI 1 μm

- RODS 1 x 2 – 4 μm

VIRUSES 0,1 μm

methylene blue staining
Methylene blue staining
  • This method distinguish live (colourless) and dead (coloured) cell.
  • Saccharomyces cerevisiae – yeast; baker, beer production
methylene blue staining1
Methylene blue staining
  • A drop of water is placed in the centre of a slide.
  • Two loopful of yeast are transferred to slide
  • One loopful of methylene blue is added
  • Examine with dry objectives
slide9

YEASTS

budding

slide11

COCCI

pediococci, tetrades

diplococci

sarcina

streptococci

staphylococci

slide12

Axis of division

diplococci

streptokoky

tetrades

sarcinas

staphylococci

slide14

RODS- regular endospore-forming

plektridium

Clostridium

Bacillus

slide15

RODS - curved

vibrio

spirilla

spirochaeta

slide17

RODS- irregular

bifidobacteria

mycobacteria

negative staining
Negative Staining
  • (Background staining)
  • This method consist of mixing the microorganisms in a small amount of nigrosine and spreading the mixture over the surface of a slide.
negative staining1
Negative Staining
  • Drops of water and nigrosine are placed in the centre of a microscopic slide.
  • Remove a small amount of material from between your teeth with a sterile straight toohpick.
  • Spread the mixture of water, nigrosine and sample over the slide.
  • Allow the slide to air-dry and examine with an oil immersion objective