Inquiry into Bacteria:

These are focus question to help design your Data Sheet

Topic One: Myths and Monera

Prior to making your data sheet consider what comes to your mind when you hear or read the term bacteria?

Some questions you may want to consider while reading the text:

• Where are bacteria found?
• Are most bacteria harmful?
• How are bacteria related to other organisms?
• What is the difference between a prokaryote and eukaryote
• How are the activities of life useful in examining how bacteria interact with other species?

Topic Two: The problems of classification

Here is the first thing to consider:

• What was the original system to classify organisms?
• What properties or observations did this system use for classification?
• Can these observations be used to classify microscopic organism?
• What techniques could be used to establish observational criteria?
• What are some limitations of classifying organism based upon structures?

Topic Three: Metabolic Perspectives

Now we have gone molecular, so how can metabolic pathways provide a new way to classify organism?

Consider the following

• How do organism get or make their energy?
• How do organisms feed themselves?
• How is the environment related to metabolism?

Topic Four Interactions with Humans

Positive Interactions

• What are the top ten positive interactions with bacteria?

Negative Interactions

• How is metabolism and structure related to negative interactions?
• How are negative interactions prevented?

(from a macrobiotic to molecular perspectives)

Topic Five: Playing with critters!

• What is the criteria for sanitary technique?
• How can metabolic activities be used make observations about bacteria?
• What simple tests and ideas can we create?

Topic Six: Creating a testable experiment!

• You will be responsible for creating an experiment!

Bacteria Notes:

Part One: Unity and Diversity

Ways to classify bacteria

• Shape and size
• Gram positive and Gram negative
• Metabolism (include respiration and food source)
• Archeo and Eubacteria

1. Size and Shape

• Bacteria were first described by Leeuwenhoek in 1677 after he had invented the compound microscope.
• Bacteria range in size from about 1 to 10 um long by about 0.2 to 0.3 um across (1 um = .001 mm).

Most true bacteria come in one of three different shapes:

(1) Rod shaped: Bacillus(i) [filaments or single].

(2) Spherical shaped: Coccus(i) [pairs, chains, groups or single].

(3) Spiral shaped: Spirillum(a) [seldom in colonies]

Some bacteria tend to form groups:

Diplococci are pairs of spherical shaped bacteria

Streptococci are chains of spherical shaped bacteria

Staphylococci are clusters of spherical shaped bacteria

B: Gram positive and Gram negative

The term gram positive or negative, refers to both a staining proccess and specific structure of bacterial cell membranes or wall. The gram staining method is one of the more important techniques in microbiology. Yet one has to realize that this technique is not 100 percent fool proof. Differences in results can be due to type of stains and age of bacteria.

• The staining process follows the following protocal:
• Heat fix bacteria to slide
• Stain with purple dye (crystal violet), rinse with distilled water
• Stain with with iodine (marker), rinse with water
• Rinse with alchol wash, functions as a decolorization process in which negative lose colour.
• Stain with safranin (red stain) which is counter stain for gram negative

In regards to cell membrane structures:

Gram positive bacteria have cell walls composed of peptidoglycan (murein) and teichoic acid. ( basically a sugar based structure combined with amino acids)

Gram negative bacteria also have cell walls composed of a peptidoglycan ( in small amounts) but this layer is surrounded by a lipopolysaccharide outer membrane.

Comparison of Characteristic of Gram + and Gram –

C: Metabolism and Nutrition

Cell Metabolism:

Nutrients are ingested and then:

1. broken down by enzymes within cell
2. further breakdown of material is done to produce energy

• Energy is absorbed by biochemical ADP (adenine di phosphate)
• Energy is released by biochemical ATP ( adenine tri phosphate)

Energy can be produced with or without oxygen

1. Anearobic: are bacteria that do not need oxygen for cell metabolism
2. Areorobic: are bacteria that require oxygen for cell metabolism
3. Facualtative: are bacteria that can metabolize with or without oxygen

• Obligate Aerobes are those which must have oxygen
• Obligate Anaerobes are bacteria which cannot tolerate

There are many types of nutrition found among bacteria:

Autotrophic Nutrition:

1. Some are photosynthetic (use sunlight energy to produce their own food).
2. Some are chemosynthetic (oxidize inorganic compounds to obtain their

energy to produce their own food).

• These organisms are known as Photoautotrophs and Chemoautotrophs in that they manufacture their own food.

Heterotrophic Nutition.

They must obtain their energy and nutrients from other sources.

For example:

• Saprophytic bacteria : digest materials in their environment by releasing powerful digestive enzymes. They then absorb the digested nutrients.
• Parasitic bacteria : rely on other organisms to provide the digested nutrients directly.

Part Two: Interactions

Bacteria can exist everywhere there is life this includes:

• in the air
• in the water
• in the earth
• on plants
• in organism
• without bacteria, we as humans could not exist
• bacteria are the most primitive form of life because they:

1. a) grow and reproduce ( as often as one time every 15-25 minutes)
2. b) they use nutrients to survive
3. c) they have simple cell structures

• it is possible to see bacteria through a light or electron microscope
• Bacteria are both helpful and harmful
• To preserve or stop bacteria metabolism they can be:

chilled

dried out

frozen

heated

Dangerous bacteria are called pathogens because they cause diseases.

• to control pathogenic bacteria you can remove bacteria by:

removing all bacteria using extreme heat

wash with antiseptic soaps

use antibiotics

specific immunization for specific bacteria proteins

• Some bacteria can become resistant to antibodies by altering protein coat or structure of cell wall.
• Bacteria can change into dormant forms called spores, which allow the bacteria to stop metabolism in extremely harsh environments

Useful Bacteria (at least 12 reasons!)

Most bacteria are not pathogenic — include decomposers, nitrogen

fixing bacteria, vitamin producing bacteria, bacteria used to make

insulin and growth hormone, bacteria used in dry cleaning, tanning,

cheese, yogurt

Essential Bacteria:

Bacteria can be helpful because:

• they help plants absorb nutrients from the soil ( specifically nitrogen)
• they are used to make milk products such as yogurt, cheese and butter
• they can be used to manufacture antibiotics
• they can alter biproducts from sewage treatments into non toxic waste
• they can be used to produce specific gases such as methane
• they are used for fermentation

• Escherichia coli Gram – rod shape (bacilli)
• Sarcina lutea Gram + round shape (cocci)
• Bacillus subtilis Gram + rod shape (bacilli)
• Bacillus cereus Gram + rod shape (bacilli)
• Serratia marcescens Gram – rod shape (bacilli)
• Rhodospirillum rubrum Gram – spiral shape (spirilla)

Harmful Bacteria

Harmful bacteria can cause disease (Pathogenic) by interfering with the host’s normal routine, by destroying cells and tissues, by producing endotoxins and exotoxins, and by eliciting an immune response.

• An endotoxin is a toxin within the bacterium that is only released when the bacterium dies and it breaks down.
• An exotoxin is a toxin released by living bacteria.

Koch’s Postulates: –

used to prove that an organism is responsible for a particular disease.

1. must be shown that the organism in question is always present in

the diseased hosts.

2. microbe must be isolated from the host and grown in a pure

culture.

3. microbe from pure culture must be capable of producing the

disease symptoms in a new healthy host.

4. microbes isolated from the newly infected host must be grown in a

pure culture and compared to the original micro-organism.

Examples:

1. a) Respiratory Tract: Strep throat, Rheumatic fever, Scarlet fever;

Pneumonia, Whooping cough, Diphtheria, Tuberculosis…

1. b) Skin: Staph (pimples and boils), Leprosy, Gas gangrene…
2. c) Nervous System: Tetanus, Botulism, Meningitis…
3. d) Digestive System: Typhoid fever, Cholera, Dysentery; (food

poisoning) Salmonella, Botulism, Staph…

1. e) Venereal Diseases: Gonorrhea, Syphilis…

Infection by bacteria

There are three lines of defence through which bacteria must

penetrate:

1. a) through the strong epidermal tissue (skin).. .in wounds, pores,

openings.

1. b) phagocytic white blood cells which engulf foreign materials including bacteria.
2. c) antibodies produced by other white blood cells.

Active Immunity –

– is slow acting (because the body is taking time to produce antibodies against the infection); but long lasting (because the body produces “memory cells” which “remember” how to produce these specific antibodies again).

Normally produced by actual initial infection (chicken pox), or by using a vaccine (polio, smallpox). The vaccine consists of either weakened (attenuated) bacteria, dead bacteria, or artificial products which resemble the actual foreign invader (antigen).

Passive Immunity

This is fast acting (because the person is injected with antiserum containing the necessary antibodies or antitoxin); but short-lived (because the person does not actually produce the antibodies – hence no memory cells for the future!). The protein antibodies are often provided from the blood of a larger animal such

as a horse! (Tetanus, Rabies). New techniques have been designed so that bacteria can produce specific antibodies. Newborns initially have Passive Immunity through the passage of antibodies across the placenta, and in the Mother’s milk. In some cases, injection of Toxoids stimulate the production of natural antitoxins.

Antibiotics a biological substance which will kill or slow (inhibit) the growth of an organism.

e.g. Penicillin, Tetracycline, Bacitracin, Ampicillin,

Erythromycin…

1. a) must be bacteria-specific.
2. b) some people are allergic to certain antibiotics.
3. c) some could kill off useful bacteria.
4. d) may reduce the competitive pressure and allow

harmful bacteria to survive.

1. e) may cause resistant strains to develop.

Other biocides include: Sulfur Drugs, antitoxins, various

bacteriocides, disinfectants…

Part Three: Changes with time

Reproduction

1. Bacteria reproduce mostly asexually by a process called BinaryFission. In this method, the circular ring of DNA replicates, and then the cell divides into two daughter cells — each with its own DNA.

2. Some bacteria can also undergo sexual reproduction by a process called Conjugation. In this method, the “male” is connected to the “female” by a tube called a Pilus. The DNA from the “male” then travels through the tube to the “female”. Here, it recombines with the “female” DNA and the “female” bacterium then divides.

• Some bacteria can be Transformed into a different cell by absorbing fragments of DNA of other cells. In another method of producing recombined bacterial DNA, bacteriophages (viruses) carry portions of the bacterial DNA from one cell to another. This process is called Transduction.

4. When environmental conditions are not favourable, some bacteria are capable of forming highly resistant thick-walled Endospores until conditions once again return to normal.

Part Four Form and Function

Cell Structure:

A bacteria cell has the following structures:

• Nuclear material in the form of DNA to pass on genetic information
• Cell membrane: which controls the flow of material in and out of a cell
• Ribosome: which assist in making cell proteins
• Cell wall
• Flagella
• Endospores

Bacteria Lab Questions:

1. Why must you always use sterile technique when working with bacteria?
2. What does “pathogenic” mean?
3. What is an “inoculum”?
4. List three general rules for handling bacteria.
5. When is the Pour Plate Method of culturing bacteria used?
6. Why must you be careful in regulating the temperature of the melted agar in the Pour Plate Method?
7. In procedure 7 in each of the labs, why did you have to flame the mouths of the test tubes?
8. What is the reason for using the streak plate method?
9. What is the reason for using the Pour plate method?

BIOLOGY 11 UNIT E – MICROBIOLOGY

1. PRESCRIBED LEARNING OUTCOMES

By the end of this unit, you must be able to:

1. show an understanding of characteristics and functions of viruses and bacteria.

1. Viruses

• evaluate the evidence used to classify viruses as living or non-living
• describe the structure of viruses
• describe viral reproduction
• evaluate the effects of viruses on humans

1. Monera

• analyse monerans as a lifeform at the prokaryotic level of organization
• describe the structure and function of bacteria
• describe moneran diversity
• describe the roles and effects of bacteria
• evaluate the effectiveness of various antibiotics, disinfectants, or antiseptics on bacterial cultures

1. VOCABULARY

By the end of this unit, you must be able to define the following:

True Bacteria Notes:

Part One: Unity and Diversity

Ways to classify bacteria

• Shape and size
• Gram positive and Gram negative
• Metabolism (include respiration and food source)

A. Size and Shape

• Bacteria were first described by Leeuwenhoek in 1677 after he had invented the compound microscope.

• Bacteria range in size from about 1 to 10 um long by about 0.2 to 0.3 um across (1 um = .001 mm).

Most bacteria come in one of three different shapes:

(1) Rod shaped:                             Bacillus(i)     [filaments or single].

(2) Spherical shaped:                    Coccus(i)     [pairs, chains, groups or single].

(3) Spiral shaped:                         Spirillum(a)     [seldom in colonies]

Some bacteria tend to form groups:

Diplococci                                        are pairs of spherical shaped bacteria

Streptococci                                    are chains of spherical shaped bacteria

Staphylococci                                 are clusters of spherical shaped bacteria

B: Gram positive and Gram negative

The term gram positive or negative, refers to both a staining proccess and specific structure of bacterial cell membranes or wall. The gram staining method is one of the more important techniques in microbiology. Yet one has to realize that this technique is not 100 percent fool proof. Differences in results can be due to type of stains and age of bacteria.

• The staining process follows the following protocal:

1. Heat fix bacteria to slide
2. Stain with purple dye (crystal violet), rinse with distilled water
3. Stain with with iodine (marker), rinse with water
4. Rinse with alchol wash, functions as a decolorization process in which negative lose colour.
5. Stain with safranin (red stain) which is counter stain for gram negative

In regards to cell membrane structures:

• Gram positive bacteria have cell walls composed of peptidoglycan (murein) and teichoic acid. ( basically a sugar based structure combined with amino acids)
• Gram negative bacteria also have cell walls composed of a peptidoglycan ( in small amounts) but this layer is surrounded by a lipopolysaccharide outer membrane.

Comparison of Characteristic of Gram + and Gram –

C: Metabolism and Nutrition

Cell Metabolism:

Nutrients are ingested and then:

1. broken down by enzymes within cell
2. further breakdown of material is done to produce energy

• Energy is absorbed by biochemical ADP (adenine di phosphate)
• Energy is released by biochemical ATP ( adenine tri phosphate)

Energy can be produced with or without oxygen

1. Anearobic: are bacteria that do not need oxygen for cell metabolism

2. Areorobic: are bacteria that require oxygen for cell metabolism

3. Facualtative: are bacteria that can metabolize with or without oxygen

• Obligate Aerobes are those which must have oxygen
• Obligate Anaerobes are bacteria which cannot tolerate

There are many types of nutrition found among bacteria:

Autotrophic Nutrition:

1. Some are photosynthetic (use sunlight energy to produce their own food)

.

2. Some are chemosynthetic (oxidize inorganic compounds to obtain their

energy to produce their own food).

• These organisms are known as Photoautotrophs and Chemoautotrophs in that they manufacture their own food.

Heterotrophic Nutrition.

They must obtain their energy and nutrients from other sources.

For example:

• Saprophytic bacteria : digest materials in their environment by releasing   powerful digestive enzymes. They then absorb the digested nutrients.

• Parasitic bacteria : rely on other organisms to provide the digested nutrients directly.

Part Two: Interactions

Bacteria can exist everywhere there is life this includes:

in the air

in the water

in the earth

on plants

in organism

• without bacteria, we as humans could not exist
• bacteria are the most primitive form of life because they:

1. a) grow and reproduce ( as often as one time every 15 minutes)
2. b) they use nutrients to survive
3. c) they have simple cell structures

• it is possible to see bacteria through a light or electron microscope
• Bacteria are both helpful and harmful
• To preserve or stop bacteria metabolism they can be:

chilled

dried out

frozen

heated

Dangerous bacteria are called pathogens because they cause diseases.

• to control pathogenic bacteria you can remove bacteria by:

removing all bacteria using extreme heat

wash with antiseptic soaps

use antibiotics

specific immunization for specific bacteria proteins

• Some bacteria can become resistant to antibodies by altering protein coat or structure of cell wall.

• Bacteria can change into dormant forms called spores, which allow the bacteria to stop metabolism in extremely harsh environments

Useful Bacteria

Most bacteria are not pathogenic — include decomposers, nitrogen

fixing bacteria, vitamin producing bacteria, bacteria used to make

insulin and growth hormone, bacteria used in dry cleaning, tanning,

cheese, yogurt

Essential Bacteria:

Bacteria can be helpful because:

• they help plants absorb nutrients from the soil ( specifically nitrogen)
• they are used to make milk products such as yogurt, cheese and butter
• they can be used to manufacture antibiotics
• they can alter biproducts from sewage treatments into non toxic waste
• they can be used to produce specific gases such as methane
• they are used for fermentation

Escherichia coli                              Gram – rod shape   (bacilli)

Sarcina lutea                                    Gram + round shape (cocci)

Bacillus subtilis                               Gram + rod shape   (bacilli)

Bacillus cereus                                Gram + rod shape   (bacilli)

Serratia marcescens                      Gram – rod shape   (bacilli)

Rhodospirillum rubrum                 Gram – spiral shape (spirilla)

Harmful Bacteria

Harmful bacteria can cause disease (Pathogenic) by interfering with the host’s normal routine, by destroying cells and tissues, by producing endotoxins and exotoxins, and by eliciting an immune response.

• An endotoxin is a toxin within the bacterium that is only released when the bacterium dies and it breaks down.
• An exotoxin is a toxin released by living bacteria.

Koch’s Postulates: –

used to prove that an organism is responsible for a particular disease.

1. must be shown that the organism in question is always present in

the diseased hosts.

2. microbe must be isolated from the host and grown in a pure

culture.

3. microbe from pure culture must be capable of producing the

disease symptoms in a new healthy host.

4. microbes isolated from the newly infected host must be grown in a

pure culture and compared to the original micro-organism.

Examples:

1. a) Respiratory Tract: Strep throat, Rheumatic fever, Scarlet fever;

Pneumonia, Whooping cough, Diphtheria,                                                                    Tuberculosis…

1. b) Skin: Staph (pimples and boils), Leprosy, Gas gangrene…

1. c) Nervous System: Tetanus, Botulism, Meningitis…

1. d) Digestive System: Typhoid fever, Cholera, Dysentery; (food

poisoning) Salmonella, Botulism, Staph…

1. e) Venereal Diseases: Gonorrhea, Syphilis…

Infection by bacteria

There are three lines of defence through which bacteria must

penetrate:

1. a) through the strong epidermal tissue (skin).. .in wounds, pores,

openings.

1. b) phagocytic white blood cells which engulf foreign materials including
2. c) antibodies produced by other white blood cells.

Active Immunity –

Is slow acting (because the body is taking time to produce antibodies against the infection); but long lasting (because the body produces “memory cells” which “remember” how to produce these specific antibodies again).

Normally produced by actual initial infection (chicken pox), or by using a vaccine (polio, smallpox). The vaccine consists of either weakened (attenuated) bacteria, dead bacteria, or artificial products which resemble the actual foreign invader ( biological antigen).

Passive Immunity

This is fast acting (because the person is injected with antiserum containing the necessary antibodies or antitoxin); but short-lived (because the person does not actually produce the antibodies – hence no memory cells for the future!). The protein antibodies are often provided from the blood of a larger animal such

as a horse! (Tetanus, Rabies). New techniques have been designed so that bacteria can produce specific antibodies. Newborns initially have Passive Immunity through the passage of antibodies across the placenta, and in the Mother’s milk. In some cases, injection of Toxoids stimulate the production of natural antitoxins.

Antibiotics a biological substance which will kill or slow (inhibit) the growth

of an organism.

e.g.                             Penicillin, Tetracycline, Bacitracin, Ampicillin,

Erythromycin…

1. a) must be bacteria-specific.
2. b) some people are allergic to certain antibiotics.
3. c) some could kill off useful bacteria.
4. d) may reduce the competitive pressure and allow

harmful bacteria to survive.

1. e) may cause resistant strains to develop.

Other biocides include: Sulfur Drugs, antitoxins, various

bacteriocides, disinfectants…

Part Three: Changes with time

Reproduction

1. Bacteria reproduce mostly asexually by a process called BinaryFission. In this method, the circular ring of DNA replicates, and then the cell divides into two daughter cells — each with its own DNA.

2. Some bacteria can also undergo sexual reproduction by a process called Conjugation. In this method, the “male” is connected to the “female” by a tube called a Pilus. The DNA from the “male” then travels through the tube to the “female”. Here, it recombines with the “female” DNA and the “female” bacterium then divides.

1. Some bacteria can be Transformed into a different cell by absorbing fragments of DNA of other cells. In another method of producing recombined bacterial DNA, bacteriophages (viruses) carry portions of the bacterial DNA from one cell to another. This process is called Transduction.

4. When environmental conditions are not favourable, some bacteria are capable of forming highly resistant thick-walled Endospores until conditions once again return to normal.

Part Four Form and Function

Cell Structure: A bacteria cell has the following structures:

• Nuclear material      in the form of DNA to pass on genetic information
• Cell membrane:      which controls the flow of material in and out of a cell
• Ribosome:                which assist in making cell proteins
• Cell wall
• Flagella
• Endospores

Response to Stimulus:

Bacteria Lab Questions:

1. Why must you always use sterile technique when working with

bacteria?

2. What does “pathogenic” mean?

3. What is an “inoculum”?

4. List three general rules for handling bacteria.

5. When is the Pour Plate Method of culturing bacteria used?

6. Why must you be careful in regulating the temperature of the melted agar in the Pour Plate Method?

7. In procedure 7 in each of the labs, why did you have to flame the mouths of the test tubes?

8. What is the reason for using the streak plate method?

1. What is the reason for using the Pour plate method?

Designing an experiment.

• From these biological facts, could you design a lab to test a simple hypothesis?
• After you have a simple hypothesis…design a simple lab that can be done in less than one week. It should include both quantitative and qualitative data.

Inquiry into Bacteria:

Topic One:    Myths and Monera

Prior to doing this assignment consider what comes to your mind when you hear or read the term bacteria?

Some questions you may want to consider while reading the text:

• Where are bacteria found?
• Are most bacteria harmful?
• How are bacteria related to other organisms?
• What is the difference between a prokaryote and eukaryote
• How are the activities of life useful in examining how bacteria interact with other species?

Topic Two:    The problems of classification

Here is the first thing to consider:

• What was the original system to classify organisms?
• What properties or observations did this system use for classification?
• Can these observations be used to classify microscopic organism?
• What techniques could be used to establish observational criteria?
• What are some limitations of classifying organism based upon structures?

Topic Three:             Metabolic Perspectives

Now we have gone molecular, so how can metabolic pathways provide a new way to classify organism?

           Consider the following

• How do organism get or make their energy?
• How do organisms feed themselves?
• How is the environment related to metabolism?

Topic Four    Interactions with Humans

           Positive Interactions

• What are the top ten positive interactions with bacteria?

           Negative Interactions

• How is metabolism and structure related to negative interactions?
• How are negative interactions prevented?

(from a macrobiotic to molecular perspectives)

Topic Five:    Playing with critters!

• What is the criteria for sanitary technique?
• How can metabolic activities be used make observations about bacteria?
• What simple tests and ideas can we create?

Topic Six:      Creating a testable experiment!

• You will be responsible for creating an experiment!

Refernces for all these questions can be found in:

Bacteria Notes