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AGRICULTURAL AND ENVIRONMENTAL MICROBIOLOGY Department of Microbiology, Nutrition and Dietetics

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AGRICULTURAL AND ENVIRONMENTAL MICROBIOLOGY Department of Microbiology, Nutrition and Dietetics Course syllabus laboratory exercises Lecturer: Prof. Vojt ch Rada – PowerPoint PPT presentation

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Title: AGRICULTURAL AND ENVIRONMENTAL MICROBIOLOGY Department of Microbiology, Nutrition and Dietetics


1
AGRICULTURAL AND ENVIRONMENTAL
MICROBIOLOGYDepartment of Microbiology,
Nutrition and Dietetics
  • Course syllabus laboratory exercises
  • Lecturer Prof. Vojtech Rada
  • Office No. 221
  • rada_at_af.czu.cz

2
Course syllabus laboratory exercises
  • Microscopy of bacteria Negative staining, simple
    staining
  • Microscopy of bacteria Gram staining
  • Yeast study Methylene blue staining
  • Mould study Zygomycetes (Mucor, Rhizopus),
    Ascomycetes (Eupenicillium) and Deuteromycetes
    (Penicillium, Aspergillus. Asexual reproduction
    (conidiospores and sporangiospores)
  • Actinomycetes and antibiotics

3
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)

4
LABORATORY SAFETY
  • Do not drink, eat and smoke
  • Protective clothing
  • Aseptic technique
  • Bacteriological loop, needle
  • Bunsen burner
  • Bacteriological stains

5
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.

6
PROTOZA 100 µm MOULDS AND YEASTS 5 10
µm BACTERIA - COCCI 1 µm - RODS 1 x
2 4 µm VIRUSES 0,1 µm
7
Methylene blue staining
  • This method distinguish live (colourless) and
    dead (coloured) cell.
  • Saccharomyces cerevisiae yeast baker, beer
    production

8
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

9
YEASTS
budding
10
Bacteria
11
COCCI
pediococci, tetrades
diplococci
sarcina
streptococci
staphylococci
12
Axis of division
diplococci
streptokoky
tetrades
sarcinas
staphylococci
13
RODS regular - nonsporing


14
RODS- regular endospore-forming
plektridium
Clostridium
Bacillus
15
RODS - curved
vibrio
spirilla
spirochaeta
16
SPIROCHAETA
VIBRIO
17
RODS- irregular
bifidobacteria
mycobacteria
18
Actinomycetes
19
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.

20
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
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