CHAPTER 6 • Sulfur

Microbial Growth
– In amino acids, thiamine, biotin
– Most bacteria decompose proteins
Growth of Microbes
– Some bacteria use SO42 or H2S
• Increase in number of cells, not cell size
• Phosphorus
• One cell becomes colony of millions of cells
– In DNA, RNA, ATP, and membranes
• Control of growth is important for
– PO43 is a source of phosphorus
- infection control
- growth of industrial and biotech • Trace Elements
organisms
• Inorganic elements required in small
Factors Regulating Growth amounts
• Nutrients – Usually as enzyme cofactors
• Environmental conditions: temperature, pH, • Organic Growth Factors
osmotic pressure
– Organic compounds obtained from
• Generation time the environment
Chemical Requirements – Vitamins, amino acids, purines,
pyrimidines
• #1 = water!
• Elements
The Nutritional Diversity of Prokaryotes
- C (50% of cell’s dry weight) HONPS
- Trace elements
• Organic
– Source of energy (glucose)
– Vitamins (coenzymes)
– Some amino acids, purines and
pyrimidines
Nutritional Categories
• Carbon sources
– CO2 = autotroph
Nutritional Categories
– organic = heterotroph
Saprobe – lives on organic matter of dead organisms
• Energy sources
Parasite – lives on organic matter of living host =
– sunlight = phototroph pathogens
– organic = chemotroph Environmental Factors Influencing Growth
• Nitrogen • Temperature
– In amino acids, proteins • O2
– Most bacteria decompose proteins • pH
– Some bacteria use NH4+ or NO3 • Osmotic Pressure
– A few bacteria use N2 in nitrogen • Others: radiation, atmospheric pressure
fixation
Temperature Optima • Pregnant women may suffer miscarriage
Psychrophiles: cold-loving • Listeriosis:
Mesophiles: moderate temperature-loving • 2500 cases/yr
Thermophiles: heat-loving • 500 fatal
• Each has a minimum, optimum, and • Prevention
maximum growth temperature
• Pasteurization
Psychrophiles
• Avoidance
- Grow between 0°C and 15°C
Oxygen Requirements
- Found in the ocean and polar ice caps
• Obligate aerobes – require O2
Psychrotrophs
• Facultative anaerobes – can use O2 but also
- Grow between 0°C and 20-30°C
grow without it
- Cause food spoilage
• Obligate anaerobes – die in the presence of
O2
Oxygen Tolerance
• Aerotolerant – do not use O2 but can grow
when it is present
– Often ferment glucose to lactic acid
• Microaerophiles – require O2 but grow only
in concentrations lower than air
Oxygen Concentration

Temperature Optima Need Prefer Ignore Oxygen

< 2is– 10%
Oxygen Oxygen Oxygen ToxicOxygen
• Optimum growth temperature is usually near
the top of the growth range
• Death above the maximum temp. comes
enzyme inactivation
• Mesophiles most common group of
organisms
• 40ºF (5°C) slows or stops growth of most
microbes Toxic Forms of Oxygen
• Singlet oxygen (O2) – very reactive

Listeria monocytogenes • Superoxide free radicals (O2.)

• Gram + rod – Neutralized by superoxide dismutase

(SOD)
• Common in environment
• Peroxide anions (O2-2)
• Lives in monocytes (WBC)
– H2O2 broken down by catalase and
• Intracellular peroxidase
• Can move through cell membrane to spread • Hydroxyl radical (OH-) –very reactive
from cell to cell
• PSYCHROTROPH
• Listeriosis: fever, aches, GI or CNS symptoms
 • Many bacteria and viruses survive low pH of
stomach to infect intestines
• Helicobacter pylori lives in stomach under
mucus layer

Brewer’s Jar

Osmotic Pressure
– Hypertonic environments, increase salt or
sugar, cause plasmolysis
– Extreme or obligate halophiles require high
osmotic pressure
– Facultative halophiles tolerate high osmotic
pressure
Drying and High Osmolarity

Candle jar • Salted fish, jerky, honey, sweetened

condensed milk are preserved by pulling
water out of bacteria
• Hypotonic medium (low osmolarity) may lyse
bacteria without cell walls
Ecological Associations
• Symbiotic: close nutritional relationship
– Mutualism: both benefit
– Commensalism: commensal benefits,
host not harmed
– Parasitism: parasite benefits, host
harmed

• Provides low O2, high CO2

pH
• Most bacteria grow between pH 6.5 and 7.5
• Acid (below pH 4) good preservative for
pickles, sauerkraut, cheeses
• ACIDOPHILES can live at low pH
Measuring Bacterial Growth Generation Time
• Time required for cell to divide/for
population to double
Bacterial Division
• Average for bacteria is 1-3 hours
• Bacteria divide by binary fission
• E. coli generation time = 20 min
• Alternative means
– 21 generations (7 hours), 1 cell
– Budding becomes 1 million cells!
– Conidiospores (filamentous bacteria)
– Fragmentation

Generation Time

Cf = Ci2n
Cf = final cell number
Ci = initial cell number
n= no. generation
gt = .301t
log Cf-log Ci

Bacterial Growth Curve

 Measurement of Cell Numbers
• Direct cell counts
– counting chambers
– electronic counters
– on membrane filters
• Viable cell counts
– plating methods
– membrane filtration methods

 Counting chambers
o easy, inexpensive, and quick

o useful for counting both eucaryotes

and procaryotes
o cannot distinguish living from dead
cells

Phases of Growth
• Lag phase – making new enzymes in
response to new medium
• Log phase – exponential growth
– Desired for production of products
– Most sensitive to drugs and radiation
during this period
• Stationary phase
– nutrients becoming limiting or waste
products becoming toxic
– death rate = division rate
• Death phase – death exceeds division

Measuring Growth  Electronic counters

• Direct methods – count individual cells o cannot distinguish living from dead
• Indirect Methods – measure effects of cells
bacterial growth (cell mass, metabolic o quick and easy to use
increase)
o useful for large microorganisms and
Measurement of Microbial Growth
blood cells, but not procaryotes
• can measure changes in number of cells in a
 Direct counts on membrane filters
population
• can measure changes in mass of population
 o cells filtered through special – useful if amount of substance in each
membrane that provides dark cell is constant
background for observing cells
• turbidometric measures (light scattering)
o cells are stained with fluorescent dyes
– quick, easy, and sensitive
o useful for counting bacteria

o with certain dyes, can distinguish

living from dead cells
 Plating methods
o simple and sensitive

o widely used for viable counts of

microorganisms in food, water, and
soil
o inaccurate results obtained if cells
clump together
 Membrane filtration methods

Metabolic Activity

Dry Weight

 especially useful for analyzing aquatic

samples
Measurement of Cell Mass
• dry weight
– time consuming and not very
sensitive
• quantity of a particular cell constituent
– e.g., protein, DNA, ATP, or chlorophyll