Microbial Nutrition, Ecology, and Growth

Chapter 7

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Microbial Must Obtain Nutrients from Environment

7.1 Microbial Nutrition

All living things require a source of elements Essential Nutrient: any substances that must be provided to an organism Nutrients are processed and transformed into the chemicals of the cell after absorption Can also categorize nutrients according to C content

Inorganic nutrients: A combination of atoms other than C and H

Organic nutrients: Contain C and H, usually the products of living things

Chemical Analysis of Microbial Cytoplasm

Sources of Essential Nutrients

Carbon sources Nitrogen sources

Oxygen sources
Hydrogen sources

Phosphorus sources
Sulfur sources Others

Carbon Sources
The majority of C compounds involved in normal structure and metabolism of all cells are organic

Heterotroph: Must obtain C in organic form (nutritionally dependent on other living things)
Autotroph: (self-feeder) Uses inorganic CO2 as its carbon source (not nutritionally dependent on other living things)

Nitrogen Sources
Main reservoir- N2 Primary nitrogen source for heterotrophsproteins, DNA, RNA Some bacteria and algae utilize inorganic nitrogenous nutrients Small number can transform N2 into usable compounds through nitrogen fixation Regardless of the initial form, must be converted to NH3 (the only form that can be directly combined with C to synthesize amino acids and other compounds)

Oxygen Sources
Oxygen is a major component of organic compounds

Also a common component of inorganic salts

O2 makes up 20% of the atmosphere

Hydrogen Sources
Hydrogen is a major element in all organic and several inorganic compounds

Performs overlapping roles in the biochemistry of cells:

Maintaining pH Forming hydrogen bonds between molecules Serving as the source of free energy in oxidation-reduction reactions of respiration

Phosphorus (Phosphate) Sources

Main inorganic source of phosphorus is phosphate (PO43-)

Derived from phosphoric acid

Found in rocks and oceanic mineral deposits Key component in nucleic acids Also found in ATP Phospholipids in cell membranes and coenzymes

Sulfur Sources
Widely distributed throughout the environment in mineral form

Essential component of some vitamins

Amino acids- methionine and cysteine

Other Nutrients Important in Microbial Metabolism

Potassium- protein synthesis and membrane function Sodium- certain types of cell transport

Calcium- stabilizer of cell walls and endospores

Magnesium- component of chlorophyll and stabilizer of membranes and ribosomes Iron- important component of cytochrome proteins Zinc- essential regulatory element for eukaryotic genetics, and binding factors for enzymes Cooper, cobalt, nickel, molybdenum, manganese, silicon, iodine, and boron- needed in small amounts by some microbes but not others

Growth Factors: Essential Organic Nutrients

Growth factor: An organic compound such as an amino acid, nitrogenous base, or vitamin that cannot be synthesized by an organism and must be provided as a nutrient. For example, many cells cannot synthesize all 20 amino acids so they must obtain them from food (essential amino acids).

Microbial Nutritional Strategies

Carbon source: Autotroph: self-feeders use carbon dioxide

Heterotroph: other-feeders use organic carbon

Energy source: Chemotroph: use organic molecules Phototroph: use light

Lithotroph: use inorganic molecules like H2S

Every combination is possible and does exist

Nutritional Categories

Saprobes
Free-living microorganisms Decomposers of plant litter, animal matter, and dead microbes Most have rigid cell wall, so they release enzymes to the extracellular environment and digest food particles into smaller molecules Obligate saprobes- exist strictly on dead organic matter in soil and water

Other Chemoheterotrophs
Parasites Derive nutrients from the cells or tissues of a host

Also called pathogens because they cause damage to tissues or even death
Ectoparasites- live on the body Endoparasites- live in organs and tissues Intracellular parasites- live within cells

Obligate parasites- unable to grow outside of a living host

Nutrient Transport
Most nutrients are polar Do not cross the membrane alone

Requires a carrier
Need to concentrate essential nutrients

Requires energy

The Movement of Molecules: Diffusion

Diffusion: When atoms or molecules move in a gradient from an area of higher concentration to an area of lower concentration

Will eventually evenly distribute the molecules

Simple or passive diffusion is limited to small nonpolar molecules or lipid soluble molecules

The Movement of Water: Osmosis

Osmosis: Diffusion of water through a selectively permeable membrane

Membrane: is selectively permeable; allows free diffusion of water but can block certain other dissolved molecules
When solute is not diffusible, water will diffuse at a fast rate from the side that has more water to the side that has less water.

Osmosis

How Osmosis Works

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Osmotic Relationships
Relative concentrations of the solutions on either side of the cell membrane Isotonic: The environment is equal in solute concentration to the cells internal environment No net change in cell volume Generally the most stable environment for cells Hypotonic: The solute concentration of the external environment is lower than that of the cells internal environment Net direction of osmosis is from the hypotonic solution into the cell Cells without cell walls swell and can burst Hypertonic: The environment has a higher solute concentration than the cytoplasm Will force water to diffuse out of a cell Cell will shrink

Osmosis and Cells

Adaptations to Osmotic Variations in the Environment

Example: fresh pond water- hypotonic conditions Bacteria- cell wall protects them from bursting

Amoeba- a water vacuole moves excess water out of the cell

Example: high-salt environment- hypertonic conditions Halobacteria living in the Great Salt Lake- absorb salt to make their cells isotonic with the environment

Concept Check
What are the osmotic conditions in jellies and jams compared to the bacterial cytoplasm?

A. Jelly is hypertonic to the cytoplasm

B. Jelly is isotonic to the cytoplasm C. Jelly is hypotonic to the cytoplasm

Facilitated Diffusion
Used to transport hydrophilic molecules
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Facilitated Diffusion High

Extracellular

Protein carrier Movement down the concentration gradient Specificity

Saturation
Concentration

No energy required

Intracellular

Facilitated Diffusion

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Active Transport
Protein carrier and energy required Movement against the gradient
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Membrane

Membrane

Membrane

Protein Protein

Protein Protein

Protein Protein

Extracellular (a)

Intracellular

Extracellular

Intracellular

Extracellular

Intracellular

Membrane

Membrane

Protein Protein

Protein
Protein

Extracellular (b)

Intracellular

Extracellular

Intracellular

Active Transport

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Endocytosis and Exocytosis

Endocytosis- particles are engulfed Phagocytosis- process carried out by white blood cells to engulf cells or particles Pinocytosis- liquids entering the cell Exocytosis: package and release of substances from a cell

Endo- and Exocytosis

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Concept Check
Facilitated diffusion ____________.

A. requires a carrier and energy

B. requires a carrier by not energy

C. requires neither a carrier nor energy D. modifies the substrate during transport

7.2 Environmental Factors that Influence Microbes - Temperature

The range of temperatures for the growth of a given microbial species can be expressed as three cardinal temperatures: Minimum temperature: the lowest temperature that permits a microbes continued growth and metabolism Maximum temperature: The highest temperature at which growth and metabolism can proceed Optimum temperature: A small range, intermediate between the minimum and maximum, which promotes the fast rate of growth and metabolism Some microbes have a narrow cardinal range while others have a broad one

Another way to express temperature adaptation- to describe whether an organism grows optimally in a cold, moderate, or hot temperature range

Temperature Optima
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Optimum

Rate of Growth

Psychrophile Psychrotroph Thermophile Mesophile Extremethermophile

minimum

maximum

-15 -10 -5

5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 105110115120 125130

Temperature C

Psychrophile
A microorganism that has an optimum temperature below 15C and is capable of growth at 0C. True psychrophiles are obligate with respect to cold and cannot grow above 20C.

Psychrotrophs or facultative psychrophiles- grow slowly in cold but have an optimum temperature above 20C.

Red Snow- Psychrophile

Mesophile
An organism that grows at intermediate temperatures

Optimum growth temperature of most: 20C to 40C

Temperate, subtropical, and tropical regions Most human pathogens have optima between 30C and 40C

Thermophile
A microbe that grows optimally at temperatures greater than 45C

Vary in heat requirements

General range of growth of 45C to 80C Hyperthermophiles- grow between 80C and 120C

Environmental Factors- Gas

Atmospheric gases that most influence microbial growth- O2 and CO2 Oxygen gas has the greatest impact on microbial growth As oxygen enters into cellular reactions, it is transformed into several toxic products

Most cells have developed enzymes that go about scavenging and neutralizing these chemicals
Superoxide dismutase Catalase

Several General Categories of Oxygen Requirements

Aerobe: can use gaseous oxygen in its metabolism and possesses the enzymes needed to process toxic oxygen products Obligate aerobe: cannot grow without oxygen Facultative anaerobe: an aerobe that does not require oxygen for its metabolism and is capable of growth in the absence of it Microaerophile: does not grow at normal atmospheric concentrations of oxygen but requires a small amount of it in metabolism

Gas Requirements
Anaerobe: lacks the metabolic enzyme systems for using oxygen in respiration Strict or obligate anaerobes: cannot tolerate any free oxygen in the immediate environment and will die if exposed to it. Aerotolerant anaerobes: do not utilize oxygen but can survive and grow to a limited extent in its presence

Carbon Dioxide
All microbes require some carbon dioxide in their metabolism Capnophiles grow best at a higher CO2 tension than is normally present in the atmosphere

Effects of pH
Majority of organisms live or grow in habitats between pH 6 and 8

Acidophiles, Neutrophiles, Alkaliphiles

Obligate acidophiles Euglena mutabilis- alga that grows between 0 and 1.0 pH Thermoplasma- archae that lives in hot coal piles at a pH of 1 to 2, and would lyse if exposed to pH 7

Osmotic Pressure
Most microbes live either under hypotonic or isotonic conditions

Osmophiles- live in habitats with a high solute concentration

Halophiles- prefer high concentrations of salt Obligate halophiles- grow optimally in solutions of 25% NaCl but require at least 9% NaCl for growth

 Nonphotosynthetic microbes tend to be damaged by the toxic oxygen products produced by contact with light

Miscellaneous Environmental Factors

Other types of radiation that can damage microbes are ultraviolet and ionizing rays
Barophiles: deep-sea microbes that exist under hydrostatic pressures ranging from a few times to over 1,000 times the pressure of the atmosphere All cells require water- only dormant, dehydrated cells tolerate extreme drying

Concept Check
What sort of microbe only grows in the presence of oxygen?

A. Anaerobe B. Facultative anaerobe C. Aerobe

D. Halophile

Ecological Associations Among Microorganisms

Most microbes live in shared habitats. Interactions can have beneficial, harmful, or no particular effects on the organisms involved. They can be obligatory or nonobligatory to the members. They often involve nutritional interactions.

Symbiosis
A general term used to denote a situation in which two organisms live together in a close partnership Mutualism: when organisms live in an obligatory but mutually beneficial relationship Commensalism: the member called the commensal receives benefits, while its coinhabitant is neither harmed nor benefited Satellitism: when one member provides nutritional or protective factors needed by the other Parasitism: a relationship in which the host organism provides the parasitic microbe with nutrients and a habitat

Satellitism

Nonsymbiotic Relationships
Synergism an interrelationship between two or more freeliving organisms that benefits them but is not necessary for their survival Antagonism

an association between free-living species that arises when members of a community compete

Biofilms
Estimated to contribute to 80% of chronic infections Resistant to most antibiotic treatments

Mixed communities of organisms

Quorum sensing

Interrelationships Between microbes and Humans

Normal microbiotia: microbes that normally live on the skin, in the alimentary tract, and in other sites in humans Can be commensal, parasitic, and synergistic relationships

7.3 The Study of Microbial Growth

Growth takes place on two levels Cell synthesizes new cell components and increases in size The numger of cells in the population increases The Basis of Population Growth: Binary Fission

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Cell wall

Binary Fission
1 A young cell.

Cell membrane Chromosome 1 Chromosome 2 Ribosomes

Asexual process

Within 24 hours, one cell can result in

4.7 x 1021 cells 5,100 tons

2 Chromosome is replicated and new and old chromosomes move to different sides of cell.

3 Protein band forms in center of cell.

4 Septum formation begins.

5 When septum is complete, cells are considered divided. Some species will separate completely as shown here, while others remain attached, forming chains or doublets, for example.

Bacterial Cell Cycle

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The Rate of Population Growth

Generation or doubling time: The time required for a complete fission cycle Each new fission cycle or generation increases the population by a factor of 2 The length of the generation time- a measure of the growth rate of an organism
Average generation time- 30 to 60 minutes under optimum conditions
Can be as short as 10 to 12 minutes

This growth pattern is termed exponential

Graphing Bacterial Growth

The data from growing bacterial populations are graphed by plotting the number of cells as a function of time

If plotted logarithmically- a straight line

If plotted arithmetically- a constantly curved slope To calculate the size of a population over time: = (Ni)2n Nf

Nf is the total number of cells in the population at some point in the growth phase Ni is the starting number N denotes the generation number

Population Growth

Concept Check
If you have 100 bacteria with a doubling time of 20 minutes in media, how many cells would there be in two hours under optimal growth conditions?

A. 600
B. 1,200 C. 3,200 D. 6,400

The Population Growth Curve

A population of bacteria does not maintain its potential growth rate and double endlessly A population displays a predictable pattern called a growth curve The method to observe the population growth pattern: Place a tiny number of cells in a sterile liquid medium

Incubate this culture over a period of several hours

Sampling the broth at regular intervals during incubation

Plating each sample onto solid media

Counting the number of colonies present after incubation

Stages in the Normal Growth Curve

Lag Phase Exponential Growth Phase

Stationary Growth Phase

Death Phase

Growth Curve

Lag Phase
Relatively flat period Newly inoculated cells require a period of adjustment, enlargement, and synthesis The cells are not yet multiplying at their maximum rate

The population of cells is so sparse that the sampling misses them

Length of lag period varies from one population to another

Exponential Growth (Logarithmic or log) Phase

When the growth curve increases geometrically Cells reach the maximum rate of cell division Will continue as long as cells have adequate nutrients and the environment is favorable

Stationary Growth Phase

The population enters a survival mode in which cells stop growing or grow slowly

The rate of cell inhibition or death balances out the rate of multiplication
Depleted nutrients and oxygen Excretion of organic acids and other biochemical pollutants into the growth medium

Death Phase
The curve dips downward Cells begin to die at an exponential rate

Potential Importance of the Growth Curve

Implications in microbial control, infection, food microbiology, and culture technology

Growth patterns in microorganisms can account for the stages of infection

Understanding the stages of cell growth is crucial for working with cultures In some applications, closed batch culturing is inefficient, and instead, must use a chemostat or continuous culture system

Methods of Analyzing Population Growth

Turbidometry- a tube of clear nutrient solution becomes turbid as microbes grow in it

Use spectrophotometer

Enumeration of Bacteria
Direct or total cell count- counting the number of cells in a sample microscopically

Uses a special microscope slide (cytometer)

Used to estimate the total number of cells in a larger sample

Automated Counting
Coulter counter- electronically scans a culture as it passes through a tiny pipette

Flow cytometer also measures cell size and differentiates between live and dead cells
Real-time PCR allows scientists to quantify bacteria and other microorganisms that are present in environmental or tissue samples without isolating or culturing them

Automated Counting

Concept Check
Suppose that you have a suspension that contains both live and dead microbial cells. What method would be best to determine the number of live?

A. Cytometer
B. Coulter counter C. Spectrophotometer D. Colony counts after dilution