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Do yourself a favor. Use the…

Virtual Microbiology
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including:
• practice test questions
• review questions
• study guides and learning objectives
• photos of bacterial colonies, stains, media and other
lab-related material

You can access the VMC through Moodle, or by going to www.ScienceProfOnline.com

 Microbial
Growth
Lecture 8

Image: Pearson Education Inc. (2004) publishing as Benjamin Cummings

 Life and Metabolism
• All living organisms obtain
___________(_________)
from their environment.

• Nutrients are needed as

building materials for the cell
and as a source of energy to
do cellular work.

• Nutrients are metabolized

(broken down) into simpler
molecules and
_____________.

GlowingColonyEColi : A false-colored image from fluorescence microscopy of a growing colony of E coli cells. "Aging and
Death in E. coli" (2005) PLoS Biol 3(2): e58 doi:10.1371/journal.pbio.0030058.
 Microbial Growth
• Refers to increase in the
____________ of microbes
(reproduction) rather than an
increase in ____ of the
microbe.

• Result of microbial growth is

the ________ = aggregation
of cells arising from single
parent cell.

• The time required for growth

and reproduction is known as
the doubling or ___________
______.
GlowingColonyEColi : A false-colored image from fluorescence microscopy of a growing colony of E coli cells. "Aging and
Death in E. coli" (2005) PLoS Biol 3(2): e58 doi:10.1371/journal.pbio.0030058.
 Growth of Microbial Populations

Image: Pearson Education Inc. (2004) publishing as Benjamin Cummings

 ___________________ in Cell Count
From Binary Fission

Generation Number Cell Count

0 1
1 2
2 4
3 8
4 16
5 32
10 1,024
20 1,048,576

View: http://www.cellsalive.com/qtmovs/ecoli_mov.htm
Image: GlowingColonyEColi : A false-colored image from fluorescence microscopy of a growing colony of E coli cells.
"Aging and Death in E. coli" (2005) PLoS Biol 3(2): e58 doi:10.1371/journal.pbio.0030058.
 The Population Growth Curve

Image: Pearson Education Inc. (2004) publishing as Benjamin Cummings

 Generation Time Under Optimal
Conditions (at 37oC)
Organism Generation
Time (min)
Bacillus cereus 28

Escherichia coli 12.5

Staphylococcus aureus (causes many infections: toxic shock syndrome one example) 27-30

Mycobacterium tuberculosis (agent of Tuberculosis) 792 – 932

Treponema pallidum (agent of Syphilis) 1,980

Mycolic Acid, Generation Time & Disease
Meet the Microbe!
TB Culture

Phylum Actinobacteria, HIGH G+C “GRAM-POSITIVE”

BACTERIA

_______________
Genus of rod-shaped, acid-fast bacteria.

The presence of mycolic acid in the cell wall gives

Mycobacteria many characteristics that defy
medical treatment.

They give the organism increased resistance to Acid-fast

chemical damage and dehydration. stain

The mycolic acid also allows the bacterium to grow

inside macrophages, effectively hiding it from the
host's immune system.

M. tuberculosis doubles population every 18-24 hours,

while M. leprae doubles population about every 14
days.
Extremely long generation time; contributes to the
chronic nature of both diseases.
Images: TBCulture : http://phil.cdc.gov/ (image #4428) CDC/Dr.
George Kubica. Wiki.
Man with
MycobacteriumTuberculosis : CDC Public Health Image Library,
Leprosy
Elizabeth White (PHIL #8433).
Leprosy : Public Domain, Wiki
 Factors Influencing Microbial Growth
Nutritional requirements
•

Oxygen requirements
•

Temperature
•

pH
•

Osmotic Pressure Staphylococcus: Janice Haney Car , CDC PHIL#10046

• This 2005 scanning electron micrograph (SEM) depicted
numerous clumps of methicillin-resistant Staphylococcus
aureus bacteria, commonly referred to by the acronym,
MRSA
 Nutrition - General

• Organisms use a variety of

nutrients for their energy
needs and to build organic
molecules and cellular
structures.

• Most common nutrients – those

containing necessary elements
such as __________ (1/2 of the
dry weight of a prokaryotic cell), oxygen,
nitrogen, and hydrogen.
SalmonellaTyphimuniumYellow :
Rocky Mountain Labs NIAID_NIH
• Microbes obtain nutrients from
variety of sources.
 Nutrition – Carbon
Four Basic Groups of Organisms
Organisms can be categorized based on energy source & carbon (organic molecule) source.

inorganic
source

Autotroph means organism can make its own food

organic
molecules

Heterotrophs break down materials that they obtain from other organisms
Image: Pearson Education Inc. (2004) publishing as Benjamin Cummings
Nutrition - Other Chemical Requirements
1. ________________
2. ________________
3. ________________
4. ________________

make up a total of 95% of the dry

weight of a bacterial cell.

• The other 5% is composed of

Calcium, Copper, Iron, Magnesium,
Manganese, Phosphorus and Iron.

• Other elements that are needed are

__trace elements__.

• These elements are needed in

extremely small amounts.

• Most trace elements can be found

dissolved in water.
 Microbes & Oxygen
• Obligate ____________ – Need oxygen to stay alive.
Aerobic respiration = Use of O2 to break down food into
useable energy.

• Obligate ____________ – Die in presence of oxygen.

O2
It is poisonous to them.
Anaerobic respiration = break down food into useable energy
without the use of O2.

• _____________ Anaerobes – Not strict aerobes or

anaerobes.
Many yeasts and enteric bacteria…Escherichia coli and
Staphylococcus aureus.

• ________________ bacteria – Require oxygen levels

lower that that found under normal atmospheric
conditions (Helicobacter pilori – found in stomach).

• ________________ Anaerobes – Don’t use oxygen,

but are not killed by it.
(Lactobacilli - This genus will make pickles from
cucumbers and cheese from milk.)

List on the board for next slide.

 Microbes & Oxygen

Aerobic and anaerobic bacteria can be identified by growing them in liquid culture:
1: Obligate aerobic bacteria gather at top of test tube to absorb maximal amount of oxygen.
2: Obligate anaerobic bacteria gather at bottom to avoid oxygen.
3: Facultative anaerobes gather mostly at the top, since aerobic respiration is most beneficial; but as
lack of oxygen does not hurt them, they can be found all along the test tube.
4: Microaerophiles gather at upper part of test tube, not at top. Require O2, but at low concentration.
5: Aerotolerant bacteria are not affected by oxygen, and they are evenly spread along the test tube.
 Using oxygen (1/2 O2) in metabolism
creates toxic waste.
Microbes that are able to use aerobic
respiration produce enzymes to detoxify
oxygen:

Catalase: H2O2 ------- H20 and 02

Superoxide dismutase (SOD): oxygen radical ----- H20 and O2

Microbes that don’t make these enzymes

cannot exist in the presence of oxygen.
 Meet the Microbe! Campylobacter jejuni
Gram negative, curved rod. Deltaprotobacteria.
Close relative of Helicobacter.
Microaerophilic bacterium.

Campylobacter infections are ____________.

Commonly found in animal feces. We catch this
from animals, particularly chickens.

Causes “food poisoning”. One of the most common

causes of human gastroenteritis in the world.

2 – 4 million cases in US per year, peaking in summer months. Usually not life-
threatening. Resolves within 24 hours – one week.

We now have issues with C. jejuni being antibiotic resistant because we put antibiotics
in animal feed.
 Microbes & Temperature
_____________

Three-dimensional shape because of the

temperature sensitive hydrogen bonds.
These bonds will usually break at higher
temperatures, and protein become
__________.
Denatured proteins lose function.

____________
Also temperature sensitive.
Become brittle if temperature is too low.

If temperature too high, lipids will be more

liquid in form.
Outside membrane cannot preserve the
integrity of the cell and it will disintegrate.
Images:
Myoglobin: Thomas Splettstoesser Wiki PublicDomain
Lipid Bylayer Cholesterol:
cellbio.utmb.edu/cellbio/membrane_intro.htm
 Effects of Temperature on Growth

40oF 77oF 95oF

Most of our plates are incubated at 37oC

(98.6oF).
Conversion C to F = 1.8xC + 32
http://www.sciencemadesimple.com/conversions.html
Image: Pearson Education Inc. (2004) publishing as Benjamin Cummings
 Categories of Microbes Based on Temperature Range

Image: Pearson Education Inc. (2004) publishing as Benjamin Cummings

 Meet the Microbe!

Psychrophilic ________: Chlamydomonas nivalis

Images: ChlamydomonasNivalis: Green algae psychrophile, www-es.s.chiba-u.ac.jp/.../snowalgae_ak.html

WatermelonSnow: Will Beback, Wiki
WatermelonSnowball: http://exviking.net/mflowers/small/Chlamydomonas_nivalis.htm
WatermelonSnowFootprint : https:/.../seki/planyourvisit/wheretoeat.htm
 Meet the Microbe! Listeria monocytogenes
Gram positive, rod-shaped psychrophile.

L. monocytogenes is widely distributed;

found in soil, water, animals, birds, insects.

Responsible for listeriosis.

Rarely pathogenic in healthy adults (mild flu-like

symptoms).

Can be lethal in pregnant women, fetuses,

newborns elderly and immune compromised,
causing meningitis or bacterimia.

Transmitted from environment (contaminated food an

water) to human, except in the case of pregnant
woman passing on to fetus.

In vulnerable populations can have a case fatality

rate of 25%.

Facultative intracellular pathogen. Triggers its own

phagocytosis.

Lysteria are very hardy. Can grow in temperatures

ranging from 39°F (the temperature of a
refrigerator) to 99°F.
Extreme _______________

Thermophiles produce
Grand Prismatic some of the bright colors
Spring, Yellowstone (the orange colors) seen in
National Park
hot springs.

How do they tolerate

the high temperatures?
• cell membranes don’t
contain fatty acids
• special enzymes fold their
DNA into special heat-stable
coils
• enzymes themselves are
heat stable with extra bonds
between amino acids.
 pH
Meet the
Microbe!

As with temperature, bacteria have minimum, Helicobacter

optimum and maximum pH ranges. pylori

__________________

• Protozoans and most bacteria have an optimum

pH range of 6.5 to 7.5.

• pH range of human organs and tissues.

• Gram-negative, microaerophilic,
________________
and acidophilic bacterium.
• Most fungi & some bacteria grow best in acid
niches.
• Can thrive in the stomach and
upper small intestines and cause
• Obligate acidophiles have to live in an acidic ulcers.
environment.
• However, many who are infected
• Acid-tolerant Microbes will survive in an acid do not show any symptoms.
environment, but do not prefer that.
• Helicobacter spp. only known
microorganisms to thrive in highly
Images: HelicobacterPylori : Electron micrograph of H. pylori possessing multiple acidic environment of stomach.
flagella. Yutaka Tsutsumi, M.D. Professor Department of Pathology Fujita Health
University School of Medicine
 pH : _________
Meet the Microbe!
Vibrio cholerae (Cholera bacteria) will
grow outside the body at a pH of 9.0.

• Gram negative, vibrio-shaped,

alkilinophile in class
Gammaproteobacteria.

• In environment, prefers warm, salty

alkaline water and many Vibrio species
can multiply in shellfish.

• Vibrio cholerae can thrive in fresh

water.
 Physical Effects of Water

• Water is important reactant in many

metabolic reactions.

• Most cells die in absence of water.

- Some have cell walls that retain
water. What genus comes to mind?
- Endospores and cysts can cease most
metabolic activity for years.
What organisms make endospores? Which make
cysts?

• Cell walls of bacteria prevent them

from exploding in a hypotonic
environment, but most bacteria are
vulnerable in hypertonic environments.
 Osmotic
Pressure

Many bacteria can be plasmolyzed by

high concentrations of solutes.

The water moves out of the bacterium

and it dies of ‘hyperosmostic shock’
(desiccation).

Photo: Journal of Bacteriology

http://jb.asm.org/content/vol188/issue10/cover.shtml

Why can you keep honey on the cupboard

for months, even years, without it spoiling?
 Osmotic Pressure & Salt Environments

Obligate ______________
• Must live in a niche of high salt
content.

• Can grow in an environment up to

30% salt.

• If placed within a freshwater

environment, they will burst and die.

____________ Halophiles
• Can survive and tolerate high salt
niches, but do not require them to
living.

Image: Staphylococcus Epidermis: CDC Public Health Image Library, Segrid McAllister (PHIL #259)
 Glycocalyx & Osmotic Pressure
Some bacteria have an additional layer
outside of the cell wall called the
glycocalyx.

One type of glycocalyx is called a

______________________ .

• glycoproteins loosely associated with

the cell wall.

• cause bacteria to adhere to solid

surfaces and help prevent the cell from
drying out

Meet the Microbe!

The slime layer of Staphylococcus
epidermidis allows it to exist on the salty
environment of the skin.
Mannitol
Salt

STRUCTURE OF MICOBIAL CELLS

 Do you want to make this class easier?

Do yourself a favor. Use the…

Virtual Microbiology
Classroom (VMC) !
Remember, every class meeting
adds pieces to the puzzle!

The VMC is full of resources to help you succeed,

including:
• practice test questions
• review questions
• study guides and learning objectives
• photos of bacterial colonies, stains, media and other
lab-related material

You can access the VMC through Moodle, or by going to www.ScienceProfOnline.com