Microbial Metabolism

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4
Applied Microbiology

• Study of practical uses of microbes in food
  processing, industrial production, and
  biotechnology

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Environmental Microbiology

• study of microbes in their natural habitats
• The study of microorganisms and their activities
  in the environment for the purpose of
  understanding their role in maintenance of the
  biosphere and their potential use in remediating
  sites that have been disturbed.

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Microbial Ecology

• Studies the interactions between microbes and
  their environments, involving living and
  nonliving components

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Organization of Ecosystems

• Biosphere thick envelope of life that surrounds
  the earths surface
• Made up of
• hydrosphere (water)
• lithosphere (soil)
• atmosphere (air)
• Maintains and creates the conditions of
  temperature, light, gases, moisture, and minerals
  required for life processes

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Energy and Nutritional Flow in Ecosystems

• Organisms derive nutrients and energy from their
  habitat.
• Food chain or energy pyramid summarizes the
  feeding levels
• producers provide the fundamental energy
  source only organisms that can produce organic
  compounds by assimilating inorganic carbon from
  the atmosphere most are photosynthetic, also
  called autotrophs
• consumers feed on other living organisms and
  obtain energy from chemical bonds primary,
  secondary, tertiary
• decomposers primarily microbes, break down and
  absorb the organic matter of dead organisms
  recycle organic matter into inorganic minerals
  and gases, mineralize nutrients

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9
Insert figure 26.3 Trophic and energy pyramid
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The Natural Recycling of Bioelements

• Processes by which bioelements and essential
  building blocks of protoplasm are recycled
  between biotic and abiotic environments
• Essential elements are cycled through biological,
  geologic, and chemical mechanisms
  biogeochemical cycles.
• Microorganisms remove elements from their
  inorganic reservoirs and convey them into the
  food web.

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Atmospheric Cycles

• Carbon cycle
• Photosynthesis
• Nitrogen cycle

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Lithospheric Cycles

• Sulfur cycle
• Phosphorous cycle

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Cycles in the Hydrosphere
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The Structure of Aquatic Ecosystems

• Lake is stratified vertically into 3 zones or
  strata
• photic zone surface to lowest limit of sunlight
  penetration
• profundal zone edge of the photic zone to lake
  sediment
• benthic zone organic debris and mud forming the
  basin
• Stratified horizontally into 2 zones
• littoral zone shoreline, relatively shallow
  water
• limnetic zone open, deeper water

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15
Insert figure 26.18 Stratification of lake
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Aquatic Communities

• Microbial distribution is associated with
  sunlight, temperature, oxygen levels, and
  available nutrients.
• Photic zone is most productive-contains plankton
• phytoplankton variety of photosynthetic algae
  and cyanobacteria
• zooplankton microscopic consumers

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• Large bodies of standing water develop thermal
  stratification.
• Epilimnion upper region, warmest
• Hypolimnion deeper, cooler
• Thermocline buffer zone between warmest and
  coolest layers

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Insert figure 26.19 Profiles of a lake
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Classification of Microorganisms
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Classification

• Over 1.5 million different organisms have been
  identified to date.
• Many similarities among living organisms
• Made up of cells surrounded by a plasma
  membrane.
• Use ATP as energy source.
• Store genetic information as DNA.
• Ribosomes are the site of protein synthesis.
• Both differences and similarities among
  organisms are caused by natural selection
  (Darwin, 1858).
• Organisms can be classified into taxonomic
  categories (taxa), based on the differences and
  similarities among them.

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Classification

• Ancient Greeks classified all living organisms
  into two groups
• Kingdom Plantae
• Kingdom Animalia
• In 1850s bacteria and fungi were incorrectly
  placed in the Plant Kingdom.
• In 1860s Kingdom Protista was proposed to
  include bacteria, fungi, algae, and protozoa, but
  many scientists still classified bacteria and
  fungi as plants.

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Classification

• In 1930s electron microscopy made it clear that
  bacterial cells lacked a nucleus. The term
  prokaryote was introduced in 1937.
• In 1959 Kingdom Fungi was established.
• In 1961 the current definition of the term
  prokaryote was established.
• In 1968 the Kingdom Prokaryote was accepted by
  biologists.
• In 1969 Robert Whitaker proposed a five-kingdom
  system of biological classification for all
  living organisms.

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• Five-Kingdom System of Biological Classification
• Proposed in 1969 by Robert Whitaker
• 1. Kingdom Prokaryotes (Monera) Oldest known
  cells. Lived over 3.5 billion years ago. Lack a
  nucleus and membrane bound organelles.
• The other four kingdoms are eukaryotes. Have a
  true nucleus and membrane bound organelles.
• 2. Kingdom Protista Mostly unicellular, lack
  tissue organization. Most have flagella during
  life.
• 3. Kingdom Fungi May be unicellular (yeasts) or
  multicellular (molds). Many are saprotrophs.
• 4. Kingdom Plantae Multicellular,
  photosynthetic.
• 5. Kingdom Animalia Multicellular, heterotrophs
  that ingest food through a mouth or oral cavity.

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Five-Kingdom Classification System
25

Prokaryotic Organisms

• Recent developments in molecular biology and
  biochemistry have revealed that there are two
  types of prokaryotic cells, based on differences
  in their ribosomes, cell walls, and metabolism.
• 1. Eubacteria True bacteria.
• Cell wall contains peptidoglycan.
• Sensitive to antibiotics.
• 2. Archaeabacteria Ancient bacteria
• Cell walls lack peptidoglycan, resistant to
  antibiotics.
• Live in extreme environments
• Three kingdoms
• 1. Methanogens Strict anaerobes that produce
  methane.
• 2. Extreme Halophiles Require high salt
  concentrations.
• 3. Thermoacidophiles Live in hot, acidic
  environments.

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Eukaryotic Organisms

• Eukaryotic Cells
• Have Complex Internal Membrane
  System          Compartmentalization         Mem
  brane-Enclosed Organelles
• DNA is enclosed in a membrane-bound nucleus
• Includes         Animal Plant
  cells        Eukaryotic Microorganisms

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Prokaryote Eukaryote

• Simpler internal structure
• Absence of nucleus
• One circular chromosome, not in a membrane
• No histones
• No membrane enclosed organelles
• Peptidoglycan cell walls
• Binary fission for cell division
• Smaller

• Contain nucleus
• Paired chromosomes, in nuclear membrane
• Histones
• Membrane enclosed organelles
• Simple (polysaccharide) cell walls
• Cell division by mitosis or meiosis
• Larger

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Structure of Prokaryotic vs. Eukaryotic cell
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Viruses

• Non cellular
• Obligate intracellular parasites
• They must live inside another cell to survive
• Have only one type of nucleic acid
• DNA or RNA (never both)
• Single or Double stranded
• Protein coat (no plasma membrane)
• Few to no enzymes
• Takes enzymes and use host cell metabolic
  machinery
• No metabolic activity
• They require a host cell to exhibit the
  characteristics of life.
• Virus diversity
• Different viruses have different hosts
• Only some viruses cause disease

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Size

• Typical prokaryote ?1 - 5 ?m long
• Typical Eukaryotic cell ?10 - 100 ?m
• Typical virus ? 50 - 80 nm

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Wednesday, February 10, 2021
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• ????????????
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  (???? 1-2- ???? 64)
• ?? ??? ?? ????? ???? ????? ? ?? CO2 ?? ????? ????
  ???? ? ?? H2O, H2 ?? H2S ?? ????? ????? ???????
  ??????? ??????.
• ????????????? ?? ???????????????
• ????? ??????????? ??????? ?? ?? ??? ?? ????? ????
  ????? ? ?? ??????? ??? ?? ????? ????? ???? ?
  ??????? ??????? ??????? ??????.
• ????????? ????????? ???????? ?????????????

33

• ?????????????? (??????????)
• ????? ?? ???? ?????????? ??????? ????? ?? ????
  NH4, NO2, H2S ? ... ? CO2 ?? ????? ???? ????
• ???? ????? ?????.
• ????????? ????????? (???? ????? ???????????? ?
  ?????????? ?? ????? NH4 ? NO2-)
• ????????? ????? (???? ????? ???????? ?? H2S ?
  ?????????? ?? ????? ?????)
• ????????? ??? (???? ????? ??????????? ?? Fe2)
• ????????? ??????? (???? ????? ????????????? ?? H2
  )
• ???????????? (??????????)
• ???? ????????? ?????? ? ?? ????? ??

34
???? ???? ??????????????? ?? ??? ???? ???? ????

• ??? ?? ??? ??? ?? ??? ????? ????????? ????.
• 1- ??? ?? ??? ???? ?????? ??????? ? ???? ???? ??
  ???? ??????? ?????????
• 2- ??? ?? ??? ???? ?????? ??????
• ?? ??? ???? ???????? ?? ??? ????? ?? ?? ????
  ????? ??????? (???? 29 wastewater bacteria)

35
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• Bacteria grow in the presence or absence of free
  molecular oxygen
• Aerobes, Anaerobes, and Facultative anaerobes
• Aerobes require oxygen for the degradation of
  substrate (filamentous organisms
  Haliscomenobacter hydrossis and Sphaerotilus
  natans, the floc former Zoogloea ramigera, and
  the nitrifying bacteria Nitrosomonas and
  Nitrobacter.
• Active aerobes are not found in anaerobic
  digesters.
• Anaerobic bacteria do not use free molecular
  oxygen for the degradation of substrates.
• Anaerobes include sulfate-reducing bacteria and
  methane-forming bacteria that use sulfate (SO42-)
  and carbon dioxide, respectively.

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Anaerobic bacteria

• There are two important groups of anaerobic
  bacteria
• Oxygen-tolerant (aerotolerant) anaerobic bacteria
  and the oxygen-intolerant (obligate) anaerobic
  bacteria.
• Obligate anaerobes such as the methaneforming
  bacteria die in the presence of free molecular
  oxygen.
• Obligate anaerobes are not killed by free
  molecular oxygen. They are killed by superoxide
  (O2-) and hydrogen peroxide (H2O2).
• Most obligate anaerobic bacteria including the
  methane-forming bacteria lack the enzymes
  superoxide dismutase and catalase and succumb to
  the toxic effects of superoxide and hydrogen
  peroxide.

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Facultative anaerobes

• Between the aerobes and anaerobes are the
  facultative anaerobes.
• Facultative anaerobic bacteria have the ability
  to use free molecular oxygen or another molecule
  to degrade substrate.
• Facultative anaerobic bacteria have the most
  complex enzyme systems with respect to
  degradation of substrate. They have one enzyme
  system for the use of free molecular oxygen and
  have another enzyme system for the use of an
  alternate molecule to degrade substrate when
  oxygen is not available.
• They prefer oxygen to other molecules such as
  nitrate (NO3-)
• Denitrifying bacteria including Bacillus,
  Escherichia, and Pseudomonas are facultative
  anaerobic bacteria.

38
???????????????? ??? ????? ?? ?? ? ??????

• ???????? (Bacteria)
• ?????? (Fungi)
• ??????????? ?? ????????(Protozoa)
• ???????? ? ???????(Rotifers and Nematodes)
• ??????(Algae)
• ???????(Viruses)

39
?????? (?????? yeast? ????? mold? ?????? mushroom)

• ???????????? ????????? ????????? ???? ??? ????? ?
  ???????????????
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  ????? ? ????? ??????
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40
Filamentous fungi. Filamentous fungi occasionally
bloom in activated sludge processes due to low pH
or nutrient deficiency. Filamentous fungi are
relatively large in size and display true
branching.
41
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42
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43
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• ????????? (??????)
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  feet)
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  ???????????)
• ??????????? (?????????)
  Mastigophora (flagellates)
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• ????????? (?????????)
  Ciliophora
  (ciliates)
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  ???????)
• ?????????
  
  Sporozoa
• ???? ????? ????? ???? ? ???? ???? ?????.
• ?????????? ?????? (?????? ??????? ?? ???? ???
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44
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  free-swimming
  ciliates
• ??? ????? ?????
  crawling
  ciliates
• ??? ????? ???? ???
  stalked
  ciliates
• ?????? ???? ??? ????? ?? ??? ????
• ?????? ??? ???? ?? ? ??? ?? ?? ????? ???? ????
• ???? ??????? ????????? ??????? ? ???? ???? ????
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• ???? ????? ???? ??? ????? ???? ??? ? ???????? ??
  ??? ???? ????????? BOD ????? ?? ???? (??? 9-8
  ???? 237 ?????)
• ?? ??????? ??? ???? ?? ????.

46
???????? ? ???????(Rotifers and Nematodes)

• ??????? ?????????? (Metazoa) ????????
• ???? ?????? ???? ?? ??? ???? ? ???? ????? ????
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• ??? ????????? ???? ?? ??? ???? ? ??? ?? ?????
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  ??????? (?????? ?? ? ???) ? ?????? ?????? (????
  ???? ???? ??? ?????? ???? ?? ?? ???? ?? ?
  ??????????)

47
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  (???? ???? ?? ??? ???? ????? ? ???? ???????? ???
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• ????? ??????? (??? ?? ????? ???? ????? ? H2O ??
  ????? ????? ???????)
• ???? ?????? ??? ?????????? ???????? ???? ? ??
  ???????? ??? (??? ????? ? ??????? ???) ?? ?????
  ???? ???? ? ?????
• ????? ??? ???? ? ???????

48
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?????? ????? ? ????? ???? ????? ?? ?????)

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• ????? ?????? ?? ????? ???? ????? ??
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• ????? ????????? (?? ????? ?????)

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• ????? ?????? ?? ????? ???? ????? ??
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• ???? ?????????? a ? d ? ???? ???????? ?????
  ???????????
• ?????? ????? ??????
• ????? ?????? ?? ????? ???? ????? ??
• ???? ???????? (??????? ???? ??)
• ???? ?????????? a ? c ? ???????????
• ?????? ????? ??????
• ????? ?????????

49
?????? ?????? ?? ????? ????

• ???? ??????? ????? ??????? ???- ??? ???? ???????
  ??????????? ?????????? ? ?????????
• ????? ???????????? ? ???????????? ? ????? ??????
  ??? ????????????

??????? ????? ?? ???????? ?????

• ???????? ??? ????????? ?????????? ??????????
  ?????? ? ????? ?? ????? ???? ??????? ????? (??
  ???? ??? ????).
• ??????? ???????????????? ????? ?? ???????? ?????
  ?? ???? ????????
• ???????? ????? ????? ?? ??????? ???? ???????
  ???-???? ??? ? ??????????
• ????? ???? 66-10 ??? ?? ??? ???? ?? ???
• ???????? ??????? ???? ????????????? ???????
  ?????? ???????? ?????????????? ??????????? ?
  ???????????
• ?????????? ?? ????? ???? ?? ??????? ???-??? ????
  ????????.

50
??? ?????? ?? ???????? ?????

• ????? ?????? ?? ??? ??? ???? ???????
  ???????????????? ????????
• ????? ??????? ?????? ???? ?? ??????? ???- ???
• ?????? ?? ?????? ??????? ? ?????? ???? ???????.
• ?????? pH ?? ??? ??????? ? ??? ?? ??? ???? ????
  ?? ?????? ?? ??????? ?? (????? ???? ????? ?????
  ????? ? ???? ??? ??????? ????? ???????)
• ????????? ???????????? ???? H2O ?? H2S ???????
  ???? ? ???? ??? ?? ???????.
• ?? ?????? ???????? ???? ????? ?????? ? ?????????
  ???????????? ???? ???????.

51
?????? ????? ?? ??????? ????? ??

• ?????? ???????? ??? ????? ?? ??? ??? ???
• ?????? ?? ? ??? (???????????? ?? ???? ???????
  ??????????? ? ??????????? ????? ?? ? ??? ?? ??
  ?????)
• ???? ???? ???????? ???? ??????? ????? ???
  ????????? ???? ?? ????? ???????? ???.
• ??????? ?? ?????? ???? ??????????? ????? ?????
  ??? ???? ??? ????
• ?????? ??????????????? ?? ?? ?????? ??
• ????? ???? ??????? ?????????? ????????????
  ?????????? ? ??????? (???? ????????????)

??? ?????? ?? ??

• ?????? ? ?????????? ? ?????? (??? ??????? ?????)
• ?????????? ???? ???? ?? ??????? (??? ???? ?????)
• ???????? ???? ?????
• ??????? ?? ??????? ??

52
???????

• ????????? ?? ???? ???? ??????
• ???????? ?????? (???????????? ??????????? ? ...)
• ???????? ????? (?????????? ? ...)
• ???????????? (?????? ? ...)
• ??????? ?? ?????? ??? ????
• ?????????? ?????? ????????? ???? ??? ??????? (???
  ?? 90 ???? ????? ?? ?????? ???? ? ?? ??? ?????
  ??????)
• ??? 90 ?? 99 ???? ??????? ?????? ?????? ??? ????
  ??????
• ??????? ?? ???????? ?? ????? ????? ??? ????
  ?????? ??? ????? ?? ?????? ??? ????
• ??? ??? ????? ?? ?????? ??? ????
• 1- ??? ???? ????? ?? ???? ?? ?? ?????? ???
• 2- ??????????? ????? ???? ????????? ??????
• 3- ????? ??? ???????? ?????? ???????????
  (?????????) ? ??????????? ???? (???????)
• ????? ?? ????? ???? ?? ???????? ?????? ??????? ??
  ??? ????? ? ???? ?????? ? pH ????? ???? ?? ???
  ?????? ?? ??? ???????.

53
??? ??????? ?? ?? ????????

• ???? ???????????? ?? ????? 3 ?? 20 ????? ?? 1000
  ???? ?? ???????? ????? ??? ?? ??????????? ???????
  ????????? ?????????? ???? ??????? ? ?????? ?????
• ??? ??????? ???? ?????? ????????? ?????????
  ?????? ? ????????? ? ???????? ? ????????

54
????????

• ??????? ?????????? ?? ?????????? ?????? (????
  ????????? ? ?????????????)
• ???? ?? ?????? ?? ???? ???? ???? ????? ? I/I
• ???????? ???? ??????? ??? ??????????????? ?
  ??????????
• ???? ????????? ?? ?????????? ?????? ????? ?????
  ????? (????? ????? biogas ?? marsh gas).
• ????????? (extremophiles) ?? ????? ??? ?? ????
  ????? 3.5 ???? (???? ?? ???? ????? ??? ???? ????
  ??? ?????? ????? ? ???? ????? ??? ?????? ???
  ????????? ??????)
• ????????? ??????? ?? ????? ???????????? ?
  ????????? ????????????? ????????? ??? ?????
  ??????.
• ?????????????? ??? ???? ?? ?????????????? ??????
  ??????.
• ????????? (????????? ?????)

55
... ????????

• ????? ???? ?? ???? ?????? (morphology)?
  ?????????? ????? ? ?????????
• ?????? ????? 0.3-3 µ ?? ?? ????? ?????????? 100 µ
  ) (Sphaerotilus natans? ???????????? (5-50 µ) ?
  ?????????? (up to 50 µ)
• ????????? ?????? ?? ?? ???? ????? ?????? ?????? ?
  ???? ??? ????? (???? ??? ?? ??? ?????)
• ???? ???????? ?? ?? ???? ???? bacillus? coccus ?
  spirillum
• ?????? ????? (80 ?? ? 20 ???? ???)(???? ???
  ???? 90 ??? ? 10 ??????)
• ????? ???? C60H87O23N12P ????? ???? C5H7O2N
• ????? ????????? ??????? ????????? (???? 3.2 ????
  25 ????)
• TRANSPORT MECHANISMS
• Diffusion
• Facilitated diffusion
• Active transport
• Motility and Nutrition

56
... ????????

• ????????? ???????? ?? ???? ???? ?? ?? ?????
  ?????? ???? ??????? (???? ????? ??? ???? ? ????
  ????? ??? ????) (???? 4.3 ???? 35 ????)
• Acetogenic bacteria (??? ?????????) (??? 4.1 ????
  35)
• Coliforms (??? ???????)
• Cyanobacteria (??? ???????????)
• Denitrifying bacteria (??? ???????)
• Fecal coliforms (??? ???????)
• Fermentative (Acid-forming) bacteria (???
  ???????)
• Filamentous bacteria (??? ???????????????)
• Gliding bacteria (??? ???????)
• Gram-negative, aerobic cocci and rods (???
  ?????????)
• Gram-negative, facultative anaerobic rods (???
  ?????????)
• Hydrolytic bacteria (??? ?????????)
• Methan-forming bacteria (??? ?????????????)
• Nitrifying bacteria (??? ????????????)

57
... ????????

• Nocardioforms (??? ????????)
• Pathogenic bacteria (??? ????????????)
• Poly-P bacteria (??? ??????????)
• Saprophytic bacteria (??? ??????????)
• Sheath bacteria (??? ????????????)
• Spirochetes (??? ??????????)
• Sulfur oxidizing bacteria (??? ??????????)
• Sulfur reducing bacteria (??? ?? ???????????)

58
?????? ??? ??????

• ??????? ???????? (???? 62 ?????)
• ????? ??????
• ????? ??? ????????
• ????? ????
• ????? ???
• ??????? ?????? (???? 64 ????? ? 65 wastewater
  bacteria)
• ???? ?????????? ?????? (???? 65 ?????)
• ?????? (?????) ???
• ????? ??? ???? ???????
• ????? ??? ????
• ????? ?????? ? ??????? ???? ?? ??? ?????? (????
  67 ?????)
• ???? ???????
• ???? ?????
• pH
• ????? ??????

59
???????? ?? ????? ????????? ??????

• ????? ?????? ?????
• ????????? (40-60)
• ???????????? (25-50)
• ?????? (8-12)
• ????? ???? ??? ????? ?? ???? ??? ?????
  ???????????? ??? ? ??????? ??? ?????? ???
• ????????? ???? ???? ????? ???? ??? ????? ??
  ??????
• BOD5 (???? 216 ???? ?????)
• TOC
• COD (????? ?????? ???? ???? ????? ???? ???? ???
  ?? CO2? H2O ? ???????) (???? 219 ???? ?????)

60
???????? ????? ????????? ??? ????

• ????? ??? ????
  Activated sludge
• ?? ??? ???? ?? ????? ??? ???? 1- ?????????? ????
  ??? ????? ????
• 
  2- ???? ????
• ????? ??? ???? ??????
  Conventional activated sludge
• ???? ?????? (???? 1500-2500 mg/L ???? ???? ?????
  ?????? ???????? ??????? ???? ????? ?? ???? ????
  ???? ???? ????? ????? ????? ?? ???? ????
  ?????????? ???? ???? ????????? ?? ????? ??????
  ??? 4 ?? 8 ????)
• ???? ?? ?????
• ?????? ???? ???? ????? (MLSS)
• ?????? ???? ???? ???? ????? (MLVSS)(65 ?? 75 ????
  MLSS)
• ???? ??? ?? ????????????? (???? 227 ?????)
• ???? ???? ?????????
• ?? ???

61
.... ???????? ????? ????????? ??? ????

• ???????? ??? ????? ?? ??????? ??? ????
• ??????? ??? ???? ???? ??????? ????????? ? ????
  ?????? ? ???
• ?????? ???? ?? 1 ?????? ?? 1000 ?????? ?? ?????
• 5 ?? 20 ???? ?? ??????? ????? ?? ???? ?? ???????
  ???? ????? ??????.
• ???????? ?? ???? ????????? ??? ???? ????? ????
  ??????? ??? ???? ?????.
• ?????? ???? ????????? ??? ???? ???? ????????
  ??????????? ?????????????? ?????????? ????????
  ???????????? ?????? ????????? ??????????
  ????????????? ??????????? ? ??? ????????????????
  ???? ?? ?? ???? ???????????? ??????? ? ??????????
• ????? ????????? ???? ??? ?? ????????? ???? ?????
  ?????? (SRB) ?? ??? ????? ??? ?????? ?????? ??
  ??????
• ????? ?? ????????? ????? ?? ??? ???? ???? 108 CFU
  ?? ??????? ???
• ???? ????????? ????????? ???? ?????? ?????
  (???????????? ? ??????????)
• ???? ?????? ?? ????? pH ?????? ???? ? ??? ???????
  (?????? ??????? ?? ??? ????????? ????????)
• ?? ????? ?? ?? ???? ??? ?????
• ????????

62
.... ???????? ????? ????????? ??? ????

• ???? CNP ???? ???????? ????? ??? ???? 10051
• ?? ????? ??? ??? ?? ??? ??????? (????? ???? ?
  ????? ????? ? ??? ?????? ?????) ? ???? F/M ?????
  (???? MLSS ????)
• ????? ???? ???? ????? ???? ?? ????? ??? ???? 3 ??
  4 ???
• ????? ???? ???? ??? ???? ???? ?????? ?? ????? ???
  (???? 243 ?????)
• SVI ??? ?? mL/g 150 ????????? ???? ???(bulking)
• ????????? ???? ?? ??? ????? ?????? ????? ??? F/M
  ?????? ???? ???? ?? ? ????? ?????? ???? ???? ??
  ???? (F/M ????? ???? ?????? ???)
• ?? ???? (foaming) ??? ?? ??? ????? ???????? ?
  ??????????

63
??????? ?????? ? ????????? ???????????? ??

• ???????? ?? ???????? ????
• ???? ??? ????????? ??????? ??????? ?????? ????.
• ?? ???? ???????????????? ???????? ???? ????? ? ??
  ????????? ???????? ?? ????????? ???? ?????? ????.
• ???? ?? ????? ?????? ???? ?? ???????? ???? ?????
  ????.
• ???? ?????? ????? ????? ?? ????????????????
  ???????? ???? ?? ????? ??????? ????? ? ?????????
  ????? ?? ??????? ????? ?? ? ?????? ????? ????.
• ????? ?? ???? ????? ????.
• ???? ?????? ?????? ????? ???? ? ????? ???? ??????
  ????.
• ???????? ???? ???? ????????? ????.

64
... ??????? ?????? ? ????????? ???????????? ??

• ???????????????? ???? ??????? ?? ????? ????
• ?????????? ??
• ?????????? ??????
• ???????????? ??????
• ?????????? ????????
• ????????????
• ?????? ? ???????????? ?????????