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Chapter 7

Microbial Nutrition,
Ecology, and Growth

Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
7.1 Microbial Nutrition
Nutrition: process by which chemical substances
(nutrients) are acquired from the environment and
used in cellular activities
Essential nutrients: must be provided to an
organism
Two categories of essential nutrients:
– Macronutrients: required in large quantities; play
principal roles in cell structure and metabolism
• Proteins, carbohydrates
– Micronutrients or trace elements: required in small
amounts; involved in enzyme function and maintenance
of protein structure
• Manganese, zinc, nickel
2
Figure 7.1 Environmental conditions that influence microbial adaptations

Sunlight supplies the basic source of


energy on earth for most organisms.
Photosynthesizers can use it directly
to produce organic nutrients that feed
other organisms. Non photosynthetic
organisms extract the energy from
chemical reactions to power cell processes.
© Courtesy: Pacific Northwest National Laboratory
Gases: the atmosphere is a reservoir
for nitrogen, oxygen, and carbon dioxide
essential to living processes.

°C K

50 320
40 310
30 300
CO2
20 290
10 280
Nutrients 0
Plant 270
litter -10
260
-20
250
-30
Soil -40 240
microbes Soil community Aquatic microbes
Organic compounds 230
© Kathy Park Talaro
Nutrients are constantly being formed Complex communities of microbes exist in nearly every place on earth. The temperature of
by decomposition and synthesis and Microbes residing in these communities must associate physically and habitats varies to a
released into the environment. Many share the habitat, often establishing biofilms and other inter relationships. significant extent
inorganic nutrients originate from among all places
non-living environments such as the pH 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 on earth, and microbes
air, water, and bedrock. exist at most points
along this wide
temperature scale.
Acidic [H+] Neutral [OH–] Basic
(alkaline)
Acid Base

The acid or base content (pH) can show extreme variations


from habitat to habitat. Microbes are the most adaptable
organisms with regard to pH.
Microbial Nutrition

• Organic nutrients: contain carbon and


hydrogen atoms and are usually the products of
living things
– Methane (CH4), carbohydrates, lipids, proteins, and
nucleic acids
• Inorganic nutrients: atom or molecule that
contains a combination of atoms other than
carbon and hydrogen
– Metals and their salts (magnesium sulfate, ferric
nitrate, sodium phosphate), gases (oxygen, carbon
dioxide) and water

4
Chemical Analysis of Cell Contents

• 70% water
• Proteins
• 96% of cell is composed of 6 elements:
– Carbon
– Hydrogen
– Oxygen
– Phosphorous
– Sulfur
– Nitrogen

• Done to understand the cell’s nutritional make-


up 5
Forms, Sources, and Functions of Essential Nutrients

• Carbon-based Nutritional Types


• Heterotroph: must obtain carbon in an organic
form made by other living organisms such as
proteins, carbohydrates, lipids, and nucleic acids
• Autotroph: an organism that uses CO2, an
inorganic gas as its carbon source
– Not nutritionally dependent on other living things

6
Growth Factors: Essential Organic Nutrients

• Organic compounds that cannot be synthesized


by an organism because they lack the genetic
and metabolic mechanisms to synthesize them
• Growth factors must be provided as a nutrient
– Essential amino acids, vitamins

7
Classification of Nutritional Types

• Main determinants of nutritional type are:


– Carbon source: heterotroph, autotroph
– Energy source:
• Chemotroph – gain energy from chemical
compounds
• Phototrophs – gain energy through
photosynthesis

8
Nutritional Categories

9
Autotrophs and Their Energy Sources
• Photoautotrophs – energy source is sunlight
– Oxygenic photosynthesis
– Anoxygenic photosynthesis
• Chemoautotrophs (lithoautotrophs) – energy source is
inorganic substances, such as hydrogen sulfide
• Methanogens, a kind of chemoautotroph, produce
methane gas under anaerobic conditions

Figure 7.2 Methanococcus


jannaschii

10
© Kathy Park Talaro
Heterotrophs and Their Energy Sources
Figure 7.3 Extracellular Digestion in Bacteria and Fungi

• Majority are chemoheterotrophs


– Aerobic respiration
• Two categories (a) Walled cell is a barrier.
Organic debris

– Saprobes: free-living
Enzymes

microorganisms that feed on


organic detritus from dead
organisms (b) Enzymes are transported outside the wall.

• Opportunistic pathogen
• Facultative parasite
– Parasites: derive nutrients from (c) Enzymes hydrolyze the bonds on nutrients.

host
• Pathogens
• Some are obligate parasites 11
(d) Smaller molecules are transported across the
wall and cell membrane into the cytoplasm.
7.3 Environmental Factors That Influence Microbes

• Niche: totality of adaptations organisms make to


their habitat
• Environmental factors affect the function of
metabolic enzymes
• Factors include:
– Temperature
– Oxygen requirements
– pH
– Osmotic pressure
– Barometric pressure

12
Adaptations to Temperature

Three cardinal temperatures:


• Minimum temperature – lowest temperature
that permits a microbe’s growth and metabolism

• Maximum temperature – highest temperature


that permits a microbe’s growth and metabolism

• Optimum temperature – promotes the fastest


rate of growth and metabolism

13
Three Temperature Adaptation Groups
Psychrophiles – optimum temperature below 15oC; capable of
growth at 0oC
Mesophiles – optimum temperature 20o-40oC; most human
pathogens
Thermophiles – optimum temperature greater than 45oC

Figure 7.9 Psychrophile


Ecological groups Optimum Mesophile
by temperature of Thermophile
adaptation

Minimum Maximum

-15-10 -5 0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 14
Temperature °C
Gas Requirements
Oxygen
• As oxygen is utilized it is transformed into
several toxic products:
– Singlet oxygen (1O2), superoxide ion (O2-), peroxide
(H2O2), and hydroxyl radicals (OH-)
• Most cells have developed enzymes that
neutralize these chemicals:
– Superoxide dismutase, catalase
• If a microbe is not capable of dealing with toxic
oxygen, it is forced to live in oxygen free habitats

15
Categories of Oxygen Requirement
• Aerobe – utilizes oxygen and can detoxify it
• Obligate aerobe – cannot grow without oxygen
• Facultative anaerobe – utilizes oxygen but can
also grow in its absence
• Microaerophilic – requires only a small amount
of oxygen
• Anaerobe – does not utilize oxygen
• Obligate anaerobe – lacks the enzymes to
detoxify oxygen so cannot survive in an oxygen
environment
• Aerotolerant anaerobes – do not utilize oxygen
but can survive and grow in its presence
16
Culturing by Oxygen Requirement
Figure 7.12 Growth medium to determine oxygen
requirements. +O2 -O2 usage by bacteria

Figure 7.11 Culturing anaerobic bacteria

Photo by Keith Weller, USDA/ARS

© Terese M. Barta, Ph.D.


Carbon Dioxide Requirement
All microbes require some carbon dioxide in their
metabolism
• Capnophile – grows best at higher CO2 tensions
than normally present in the atmosphere

Figure 7.11b. a CO2


incubator

18
Courtesy and © Becton, Dickinson and Company
Effects of pH
• Majority of microorganisms grow at a pH between
6 and 8 (neutrophiles)
• Acidophiles – grow at extreme acid pH
• Alkalinophiles – grow at extreme alkaline pH

19
Osmotic Pressure
• Most microbes exist under hypotonic or isotonic
conditions
• Halophiles – require a high concentration of salt
• Osmotolerant – do not require high
concentration of solute but can tolerate it when it
occurs

20
Miscellaneous Environmental Factors

• Barophiles – can survive under extreme


pressure and will rupture if exposed to normal
atmospheric pressure

21
7.4 Ecological Associations Among Microorganisms

Microbial Associations

Symbiotic Nonsymbiotic

Organisms live in close Organisms are free-living;


nutritional relationships; relationships not required
required by one or both members. for survival .

Mutualism Commensalism Parasitism Synergism Antagonism


Obligatory, The commensal Parasite is Members Some members
dependent; benefits; dependent cooperate are inhibited
both members other member and benefits; and share or destroyed
benefit. not harmed. host harmed. nutrients. by others.

22
Ecological Associations

• Symbiotic – two organisms


Figure 7.13 Satellitism, a type of commensalism

Staphylococcus Haemophilus
live together in a close aureus
growth
satellite
colonies

partnership
– Mutualism: obligatory,
dependent; both
members benefit
– Commensalism:
commensal member © Science VU/Fred Marsik/Visuals Unlimited

benefits, other member


neither harmed nor
benefited
– Parasitism: parasite is
dependent and
benefits; host is harmed Courtesy Arthur Hauck (Germany)

23
Interrelationships Between Microbes and Humans

• Human body is a rich habitat for symbiotic


bacteria, fungi, and a few protozoa - normal
microbial flora
• Types of relationships: commensal, parasitic,
and synergistic relationships

24
Microbial Biofilms – A Meeting Ground

• Biofilms result when organisms attach to a


substrate by some form of extracellular matrix
that binds them together in complex organized
layers
• Dominate the structure of most natural
environments on earth
• Communicate and cooperate in the formation
and function of biofilms – quorum sensing

25
Figure 7.14 Biofilm Formation and Quorum Sensing

Quorum-dependent
proteins
Chromosome

Inducer
1 molecule
5
4

Matrix 3

1 Free-swimming cells settle on a surface and remain there.


2 Cells synthesize a sticky matrix that holds them tightly to
the substrate.
3 When biofilm grows to a certain density (quorum), the cells release
inducer molecules that can coordinate a response.
4 Enlargement of one cell to show genetic induction. Inducer molecule
stimulates expression of a particular gene and synthesis of a protein
product, such as an enzyme. 26
5 Cells secrete their enzymes in unison to digest food particles.
7.5 The Study of Microbial Growth
• Microbial growth occurs at two levels: growth at
a cellular level with increase in size, and
increase in population
• Division of bacterial cells occurs mainly through
binary fission (transverse)
– Parent cell enlarges, duplicates its chromosome, and
forms a central transverse septum dividing the cell
into two daughter cells

27
The Basis of Population Growth: Binary Fission
Figure 7.15
1 A young cell at early phase of cycle

2 A parent cell prepares for division by


enlarging its cell wall, cell membrane, and
overall volume. Midway in the cell, the wall
develops notches that will eventually form
the transverse septum, and the duplicated
chromosome becomes affixed to a special
membrane site.

3 The septum wall grows inward, and the


chromosomes are pulled toward opposite
cell ends as the membrane enlarges. Other
cytoplasmic components are distributed
(randomly) to the two developing cells.

4 The septum is synthesized completely


through the cell center, and the cell
membrane patches itself so that there
are two separate cell chambers.

5 At this point, the daughter cells are divided.


Some species will separate completely as
shown here, while others will remain attached,
forming chains or doublets, for example.

Cell wall Cell membrane Chromosome 1 Chromosome 2 Ribsomes


28
The Rate of Population Growth
• Time required for a complete fission cycle is called
the generation, or doubling time
• Each new fission cycle increases the population
by a factor of 2 – exponential growth
• Generation times vary from minutes to days

Figure 7.16 *12


4500*
4000
3500

Log of 3000
( ) number Number
2500 of cells ( )
of cells
using the 11 2000
power 1500
of 2
10 1000
9 500
0 0
(b) Time
Number 1 2 4 8 16 32
of cells
Number of
1 2 3 4 5
generations
Exponential 21 22 23 24 25
value (21) (22) (222) (2222) (22222) 29
(a)
Determinants of Population Growth
The Viable Plate Count: Batch Culture Method
Flask inoculated

Samples taken at equally spaced intervals


(0.1 ml)

60 min 120 min 180 min 240 min 300 min 360 min 420 min 480 min 540 min 600 min
0.1
500 ml ml

Sample is
diluted in
liquid agar
medium
and poured
or spread
over surface
of solidified
medium
Plates are
incubated, None
colonies
are counted
Number of
colonies (CFU) <1* 2 4 7 13 23 45 80 135 230
per 0.1 ml
Total estimated
cell population <10,000 5,000 20,000 35,000 65,000 115,000 225,000 400,000 675,000 1,150,000
in flask
*Only means that too few cells are present to be assayed.
Figure 7.18 The Population Growth Curve
In laboratory studies, populations typically display a
predictable pattern over time – growth curve
Stages in the normal growth curve:
1. Lag phase – “flat” period of adjustment, enlargement;
little growth

10
Stationary phase
9
8
Logrithm (10n) of Viable Cells

7
6
5

4 The final
outcome
varies with
3 the culture.

0
5 10 15 20 25 30 35 40
Hours
Total cells in population, live and dead, at each phase
31
Few cells Live cells Dead cells (not part of count)
The Population Growth Curve
Stages in the normal growth curve:
1. Lag phase
2. Exponential growth phase – a period of maximum
growth will continue as long as cells have adequate
nutrients and a favorable environment

10
Stationary phase
9
8
Logrithm (10n) of Viable Cells

7
6
5

4 The final
outcome
varies with
3 the culture.

0
5 10 15 20 25 30 35 40
Hours
Total cells in population, live and dead, at each phase
32
Few cells Live cells Dead cells (not part of count)
The Population Growth Curve
Stages in the normal growth curve:
1. Lag phase
2. Exponential growth phase
3. Stationary phase – rate of cell growth equals rate of cell
death caused by depleted nutrients and O2, excretion of
organic acids and pollutants
10
Stationary phase
9
8
Logrithm (10n) of Viable Cells

7
6
5

4 The final
outcome
varies with
3 the culture.

0
5 10 15 20 25 30 35 40

Total cells in population, live and dead, at each phase


Hours 33
Few cells Live cells Dead cells (not part of count)
The Population Growth Curve
Stages in the normal growth curve:
1. Lag phase
2. Exponential growth phase
3. Stationary phase
4. Death phase – as limiting factors intensify, cells die
exponentially
10
Stationary phase
9
8
Logrithm (10n) of Viable Cells

7
6
5

4 The final
outcome
varies with
3 the culture.

0
5 10 15 20 25 30 35 40

Total cells in population, live and dead, at each phase


Hours 34
Few cells Live cells Dead cells (not part of count)
Other Methods of Analyzing Population Growth
• Turbidometry – most simple, use a spectrophotometer
• Degree of cloudiness, turbidity, reflects the relative
population size

Figure 7.19
Percent of light
transmitted

(1)

© Kathy Park Talaro/Visuals Unlimited (2)


(a) (b)

T ~ A or T ~ A 35