Environmental Factors that influence Microbes

- Heat
- Cold
- Gases
- Acid
- Radiation
- Osmotic Pressure
- Hydrostatic Pressure
- Other Microbes
- \

Temperature:
Cardinal Temperatures – range of temperatures for the growth of a given microbial species
- Minimum temperature -
- Maximum temperature – highest temp to tolerate for growth and metabolism
(denature enzymes) can proceed, if rises slightly above maximum, growth will stop
- Optimum temperature – temp between minimum and maximum, best range of
temperature where microorganism can grow best at fastest way

Thermophile - - extreme thermophile, organism that could be thriving up to 100+ degrees

Celsius

Psychropiles – microbes that can thrive in very cold temp and regions
Below 15 degrees Celsius
- Cannot grow above 20 degrees Celsius, it will denature their structure
- Psychrotolerant – grow slowly in the cold but have an optimum temp between 15 and
30 degrees Celsius
Mesophiles – medically significant organisms
- Individual species can grow from 10 C to 50 C
- Optimum growth temp: 20C to 40C
 - Most human pathogens 30C to 40C
- Thermoduric microbes – survive short exposure to high temperatures; common
contaminants of heated or pasteurized foods.

Thermophiles – grow optimally at temp above 45C

Live in soil and water associated with volcanic activity, compost piles, habitats directly exposed
to the sun
- General range of growth 45C TO 80c
- Extreme thermophiles grow between 80C and 121 C

Gases – oxygen has the greatest impact on microbial growth

- Microbes fall into one of the 3 categories:
- Those that use oxygen and can detoxify it
- Those that can neither use oxygen nor detoxify it (no enzymes that can detoxify aerobic
products)
- Those that do not use oxygen but can detoxify it

How Microbes process oxygen: Singlet Oxygen

As oxygen enters into cellular reactions, it is transformed into several toxic products

Singlet oxygen (O or 1O2)

- Extremely reactive molecule produed by both living and nonliving processes
- Produced by phagoycytes to kill invading bacteria
- Buildup of singlet oxygen and the oxidation of membrane lipids and other molecules can
destroy the cell

Other ways microbes process oxygen:

-Superoxide ion (O2-), hydrogen peroxide (H2O2) (needs both for bacteria to have hydroxyl
radicals) to scavenge and neutralize , and hydroxyl radicals (OH-)
- destructive metabolic by-products of oxygen
Cells use enzymes to scavenge and neutrlalize them
Step 1: Superoxide dismutase
Step 2 Catalase

Aerobe and Facultative Anaerobe:

Aerobe – use gaseous oxygen in its metabolism
- Enzymes needed to process toxic oxygen products
- Obligate aerobe: an organism that cannot grow without oxygen

Facultative anaerobe
- Does not require oxygen for its metabolism
- Capable of growth in the absence of oxygen
- Metabolizes by aerobic respiration when oxygen is present
- Adopts anaerobic metabolism (fermentation) when oxygen is absent

Microaerophile – small amount of oxygen to grow

- Does not grow at normal atmospheric concentrations of oxygen
- Requires small amount of oxygen in its metabolism
- Live in a habitat that provides a small amount of oxygen but is not directly exposed to
the atmosphere (if exposed to normal atmospheric concentration, they will die)
Anaerobe (anerobic microorganism)
- Does not utilize oxygen
- Lacks metabolic enzyme systems to detoxify oxygen products
- Strict or obligate anaerobes cannot tolerate free oxygen and will die in its presence\

Culturing Techniques for Anaerobes

- Needs specialized chamber to grow
Rubber gasket – airtight seal
- Gas generator envelope
- Anaerobic indicator strip – containing methylene blue = colorless if there is absence of
oxygen
Increases thioglycolate (reduce presence of oxygen) as you go down in test tubes
1. (distinct cloud formation)pseudonomans – bacterium that is aerobic microorganism
2. Staphylococcus aurelius – could still grow (facultative)
3. E coli (facultative)
4. Clostridium Buticerum (anaerobic) – turbid = microbial growth

Aerotolerant Anaerobes
- Do not utilize oxygen
- Can survive and grow to a limited extent in its presence
- Not harmed by oxygen because they possess alternative meechanisms for breaking
down peroxides and superoxides

Capnophiles:
- Grow best at a higher CO2 tension than is normally present in atmosphere
- Has specialized chamber that can carry higher concentration of CO2
- Incubation is carried out in a CO2 incrubator that provides 3 to 10% of CO2

pH – acidity or basicity or alkalinity of a solution

0 – 14 scale
7.0 – netural
Below 7 – acidic
Higher than 7 – alkaline solution
Obligate acidophiles: require an acidic environment for growth
- Molds and yeasts tolerate acid and are common spoilage agents of pickled foods
Alkalinophiles – live in hot pools and soils that contain high levels of basic minerals
- Bacteria that decompose urine create alkaline conditions

Osmotic Pressures – live in habitats with high solute concentration

Obligate Halophiles – can live in high sodium concentration
- Require high concentrations of salt for growth (9-25% NACL)
- Have significant modifications to their cell walls and membranes and will lyse in
hypotonic habitats

Facultative halophiles: - resistant to salt, even though they do not normally reside in high salt
environments

Radiation – Phototrophs can use visible light rays as an energy source

Protective measures against radiation:
- Yellow carotenoid pigments absorb and dismantle toxic oxygen

Protective measures against radiation:

- Yellow carotenoid pigments absorb dismantle toxic oxygen
- Other microbes use enzymes to overcome the damaging effects of UV radiation on DNA
Hydrostatic Pressure
Barophiles – deep sea microbes that exist in pressures up to 1000 times atmospheric pressure
(deepest part of the ocean)
- Strictly adapted to high pressures that they rupture when exposed to normal
atmospheric pressure

ASSOCIATION BETWEEN ORGANISMS

Symbiotic – organisms live in close nutritional relationships; required by one or both members
- Mutualism – both members benefit
- Commensalism – the commensal benefits; other member not harmed
- Parasitism – paraise is dependent and benefits; host harmed

Nonsymbiotic – organisms are free-living; relationships not required for survival

- Synergism, - members cooperate and share nutrients
- Antagonism – some members are inhibited or destroyed by others.
Symbiosis – two organisms live together in a close relationship
Mutualism - exists when organisms live in an obligatory but mutually beneficially relationship
Commensalism – benefits one member and not the other, commensal: receives benefits,
coinhabitant: neither harmed nor benefited

Nonsymbiotic Association
Antagonism – members of community compete
- Antibiosis – form of antagonism: production of inhibitory compounds, such as antibiotics
Synergism – help each other but not necessary for their survival

Biofilms – mixed communities of different kinds of bacteria and other microbes:

- Pioneer colonizer – initially attaches to a surface
- Other microbes attach to the pioneer or to the polymeric or the polymeric sugar and
protein substance secreted by the pioneer
- Quorum sensing – cells are stimulated to release chemicals for population

Generation time or doubling time

- Time required for a complete fission cycle
- Each new fission cycle doubles the population
- As long as the environment remains favorable, the doubling effect can continue at a
constant rate
- The length of the generation time is a measure of the growth rate of an organism

32 new daughter cells

Exponential growth – growth pattern of microbes
- Express population of microbes as exponents or logarithms

The population growth curve

Growth Curve: predictable pattern of growth in a population of bacteria
Viable count technique:
- Traditionally used to observe the population growth pattern
- Total number of cells is counted over a given time period
- Fundamental method of laboratory microbiology

After 4 HOURS, NEXT GENERATION STARTS (10^2)

After next 4 hours, third generation starts (10^3)

GROWTH CRUVE IN A BACTERIAL CULTURE

Lag phase – flat, after 5 hours, tail pink picture, enlargement, synthesis of formula, preparation
for their growth or division. Newly inoculated cells require a period of adjustment, enlargement
and synthesis. Cells are not yet multiplying at their maximum rate. Population of cells is so
sparse that the sampling misses them
Exponential growth phase – time where cells are actively dividing, indicated by orange test tube
color, live cells present in growth phase
Stationary phase – cells are still actively dividing, survival mode is present, dead cells organism
present, (closed system), cells mature or aging leading to death. Cells stop growing or grow
slowly. Population enters survival mode
Death cells – some cells remain present. Curve dips downward. Limiting factors intensify

Stages in Normal Growth Curve: Lag and Log phases

Practical Importance of Growth Curve:

- Antimicrobial agents rapidly accelerate death phase (disinfectants)
- Microbes in the exponential phase are more vulnerable to these agents than those in
the stationary phase
- Actively growing cells are more vulnerable to conditions that disrupt cell metabolism
and binary fission.
Growth patterns can correspond with the stages of infection:
Bacterial shedding during the early and middle stages of infection is more likely to spead it to
others
Chemostat – continuous culture system that admits a steady stream of nutrients
- Siphons off used media and old bacterial cells to stabilize growth rate and cell number
Turbidometry – tube of clear nutrient solution becomes cloudy or turbid as microbes grow in it
- Greater the turbidity, the larger the population size

Direct (total) cell count:

- Cells in a sample are counted microscopically utlizes