Fisiologi Pertumbuhan Kuman

Nataniel Tandirogang
Bagian Mikrobiologi Program Studi Kedokteran Umum
Universitas Mulawarman

Nataniel tandirogang 2007

 Ciri-Ciri Mahluk Hidup ?

Apa Yang Dimaksud Dengan

Pertumbuhban?

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 Microbial Growth
► Microbial growth = increase in
number of cells, not cell size

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 GROWTH OF BACTERIA

STATIONER PHASE
TOTAL BACTERIA

LOG OF COUNT
DECLINE PHASE

LOG PHASE VIABLE BACTERIA

Survival Phase

LAG PHASE

TIME

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 Komponen Yang dibutuhkan
• Air
• Garam-Garam Anorganik
• Mineral
• CO2
• Faktor pertumbuhan :
O2
Eh
pH
Tekanan Osmotik
Suhu
Vitamin, dll

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 The Requirements for Growth:
Chemical Requirements

► Carbon
 Structural organic molecules, energy source
 Heterotrophs use organic carbon sources
 Autotrophs use CO2

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► Nitrogen
 In amino acids, proteins
 Most bacteria decompose proteins
 Some bacteria use NH4+ or NO3
 A few bacteria use N2 in nitrogen fixation

► Sulfur
 In amino acids, thiamine, biotin
 Most bacteria decompose proteins
 Some bacteria use SO42 or H2S

► Phosphorus
 In DNA, RNA, ATP, and membranes
 PO43 is a source of phosphorus
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► Trace Elements
 Inorganic elements required in small amounts
 Usually as enzyme cofactors
 Mg, Fe, K, Ca, dll

► Organic Growth Factors

 Organic compounds obtained from the
environment
 Vitamins, amino acids, purines, pyrimidines

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 METABOLIC FUNCTIONS OF THE VITAMIN B GROUP

B VITAMIN ORGANISM REQUIRING

• THIAMINE STAPH. AUREUS

• NICOTINIC ACID L. ARABINOSUS
• RIBOFLAVIN L. CASEI
• PYRIDOXAL CL. WELCHII
• PANTHOTHENIC ACID BRUCELLA ABORTUS
• P AMINOBENZOIC ACID CL. ACETOBULYTICUM
• FOLIC ACID CL. TETANI
• BIOTIN CL. TETANI
• VITAMIN B 12 L. LACTIS

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► Oxygen (O2)

obligate Faultative Obligate Aerotolerant

Microaerophiles
aerobes anaerobes anaerobes anaerobes

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 Toxic Forms of Oxygen
► Singletoxygen: O2 boosted to a higher-
energy state
► Superoxide free radicals: O2

Peroxide anion: O22

Hydroxyl radical (OH)

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 The Requirements for Growth:
Physical Requirements
► Temperature
 Minimum growth temperature
 Optimum growth temperature
 Maximum growth temperature

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 Temperature

Nataniel tandirogang 2007 Figure 6.1

 Psychrotrophs

Nataniel tandirogang 2007 Figure 6.2

 ► pH
 Most bacteria grow between pH 6.5 and 7.5
 Molds and yeasts grow between pH 5 and 6
 Acidophiles grow in acidic environments

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► Osmotic Pressure
 Hypertonic environments, increase salt or sugar,
cause plasmolysis
 Extreme or obligate halophiles require high
osmotic pressure
 Facultative halophiles tolerate high osmotic
pressure

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 Culture Media

► Culture Medium: Nutrients prepared for

microbial growth
► Sterile: No living microbes
► Inoculum: Introduction of microbes into
medium
► Culture: Microbes growing in/on culture
medium

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 Agar

► Complex polysaccharide
► Used as solidifying agent for culture media
in Petri plates, slants, and deeps
► Generally not metabolized by microbes
► Liquefies at 100°C
► Solidifies ~40°C

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 Culture Media

► Chemically Defined Media: Exact chemical

composition is known
► Complex Media: Extracts and digests of
yeasts, meat, or plants
 Nutrient broth
 Nutrient agar

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 Culture Media

Nataniel tandirogang 2007 Table 6.2 & 6.4

 Anaerobic Culture Methods

► Reducing media
 Contain chemicals (thioglycollate or oxyrase)
that combine O2
 Heated to drive off O2

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 Anaerobic Culture Methods
► Anaerobi
c jar

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 Anaerobic Culture Methods
► Anaerobic
chamber

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 Capnophiles require high CO2
► Candle jar

► CO2-packet

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 Selective Media
► Suppress
unwanted
microbes and
encourage
desired microbes.

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 Differential Media
► Make it easy to distinguish colonies of
different microbes.

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 Enrichment Media
► Encourages growth of desired microbe
► Assume a soil sample contains a few
phenol-degrading bacteria and thousands of
other bacteria
 Inoculate phenol-containing culture medium
with the soil and incubate
 Transfer 1 ml to another flask of the phenol
medium and incubate
 Transfer 1 ml to another flask of the phenol
medium and incubate
 Only phenol-metabolizing bacteria will be
growing
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 ►A pure culture contains only one species or
strain
► A colony is a population of cells arising from
a single cell or spore or from a group of
attached cells
► A colony is often called a colony-forming
unit (CFU)

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 Streak Plate

Nataniel tandirogang 2007 Figure 6.10a, b

 Reproduction in Prokaryotes

► Binary fission
► Budding (Streptomycetacea)
► Conidiospores (actinomycetes)
► Fragmentation of filaments (H.influenzae)

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 Binary Fission

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Nataniel tandirogang 2007
 If 100 cells growing for 5 hours produced
1,720,320 cells:

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Nataniel tandirogang 2007 Figure 6.13
Nataniel tandirogang 2007 Figure 6.14
 Direct Measurements of Microbial Growth
► Plate Counts: Perform serial dilutions of a
sample

Nataniel tandirogang 2007 Figure 6.15, top portion

 Plate Count
► Inoculate Petri
plates from
serial dilutions

Nataniel tandirogang 2007 Figure 6.16

 Plate Count
► After incubation, count colonies on plates
that have 25-250 colonies (CFUs)

Nataniel tandirogang 2007 Figure 6.15

Direct Measurements of Microbial Growth
► Filtration

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 Direct Measurements of Microbial Growth
► Multiple
tube MPN
test
► Count
positive
tubes and
compare
to
statistical
MPN
table.
Nataniel tandirogang 2007 Figure 6.18b
 Direct Measurements of Microbial
Growth
► Direct Microscopic Count

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 Direct Measurements of Microbial Growth

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 Estimating Bacterial Numbers by Indirect
Methods
► Turbidity

Nataniel tandirogang 2007 Figure 620

 Estimating Bacterial Numbers by
Indirect methods
► Metabolicactivity
► Dry weight

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