Topic 6: Microbiology and Pathogens Flashcards

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1
Q

What is a microbial culture?

A

A method of multiplying microbial organisms by letting them reproduce in a predetermined culture medium under controlled lab conditions

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2
Q

What factors must be controlled when making a microbial culture?

A

Nutrient levels
Oxygen
pH
Temperature

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3
Q

What are the main problems when culturing microorganisms?

A

1 - Harmless microorganisms might mutate to dangerous ones
2 - Pathogens enter and grow - can cause disease
3 - Contamination with unwanted microorganisms will ruin the investigation

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4
Q

What precautions should be taken when culturing microorganisms?

A

Use sterile equipment
Do not remove culture from lab
Dispose of culture safely

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5
Q

What does sterile mean?

A

Being sterile is to be free from living microorganisms and their spores

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6
Q

How do you dispose of microorganism cultures safely?

A

Seal in a plastic bag and sterilise at 120 degrees for 15 minutes under high pressure and then throw it away

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7
Q

Outline the method to culture microorganisms

A
  1. Choose a microorganism to culture
  2. Obtain a culture of the microorganism
  3. Select and make up the nutrient medium
  4. Innoculate the medium
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8
Q

What are the different types of nutrient medium?

A

Liquid broth, nutrient agar and selective medium

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9
Q

What is nutrient agar?

A

A solid nutrient elly extracted from seaweed used in Petri dishes

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10
Q

How do you make nutrient agar?

A

Pour nutrient broth and molten agar into a Petri dish and let it set at 50C

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11
Q

What is the advantage of agar

A

Sets at 50C but melts at 90C so you can keep it at a high temperature

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12
Q

What is a selective medium?

A

A growth medium with a specific combination of nutrients so only a certain type of microorganism grows on it

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13
Q

Define innoculation

A

The process by which microorganisms are transferred into a culture medium under sterile conditions

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14
Q

Explain how to innoculate a liquid nutrient medium

A

Innoculating loop scrapes bacteria from solid culture to liquid nutrient broth to form innoculating broth.
Flask stoppered with cotton wool
Incubate at suitable temp
Regularly mix

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15
Q

Why do you stopper a flask of liquid culture when culturing it?

A

To prevent contamination by microorganisms in the air

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16
Q

Why do you mix a flask of liquid culture regularly when culture?

A

To ensure the broth is aerated bc microorganisms need oxygen

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17
Q

Explain how to innoculate a solid nutrient medium

A
Sterilise the innoculating loop in a Bunsen
Dip into bacteria suspension
Streak across agar surface
Replace lid, clse tape and label
Turn upside down
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18
Q

What does aseptic mean?

A

Sterile - free from contamination from harmful bacteria

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19
Q

Why are aseptic techniques used when culturing bacteria?

A

To ensure the procedure is safe and to prevent the contamination of the culture.

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20
Q

Give examples of aseptic technique

A

Using sterile equipment
Using flamed equipment
Replacing the lid of the petri dish as soon as possible

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21
Q

What is a pure culture?

A

A culture containing only one type of microorganism

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22
Q

How can a pure culture be made?

A

Making conditions aerobic/anaerobic depending on organism
Making selective medium specific to it
Reinnoculate a plate with the microorganism on it (identified by using an indicator)

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23
Q

What information do we need to measure the growth of a culture?

A

The number of cells at different times

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24
Q

Name some methods used to measure the growth of cultures

A

Cell count
Dilution plating
Turbidimetry
Size/area

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25
Q

What is a cell count?

A

A method of measuring the growth of cultures of bacteria or single selled fungi

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26
Q

What equipment is needed for a cell count?

A

Haemocytometer

Microscope

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27
Q

What is a haemocytometer?

A

A thick glass microscope slife with a rectangular chamber that holds liquid. It has perpendicular lines

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28
Q

What is trypan blue?

A

A dye that stains dead cells blue and leaves living cells white

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29
Q

Why do we dilute cells with trypan blue before putting them into a haemocytometer?

A

To stain them so counting is easier

To separate the cells to make them easier to count

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30
Q

What is the rule for cells on lines when counting cells in a haemocytometer?

A

Count them if they’re on the top or right line

Don’t count them if they’re on the bottom or left

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31
Q

How is a haemocytometer used?

A

Put under a microscope and viewed under a low power so the cells can be seen

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32
Q

Why is it important to not move the haemocytometer when counting?

A

Will mve the cells so counting becomes incorrect

Air bubble may be introduced

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33
Q

What is turbidity?

A

The cloudiness of a liquid caused by a large number of individual particles in the solution

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34
Q

What is turbidimetry?

A

A method of measuring the concentration of a substance by measuring the amount of light that passes through it. A specialised form of colorimetry

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35
Q

What are calibration curves?

A

Graphs of known concentrations against their absorbances

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36
Q

How are calibration curves made?

A

Control culture is made and samples taken at regular intervals
Put into colorimeter and turbidity is measured
Do cell count of the same sample
Repeat over time
Plot graph of turbidity against cell count and compare unknown turbidities to get cell count directly

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37
Q

How is turbidity measured?

A

The absorbance of light of a sample is measured by a colorimeter

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38
Q

What is dilution plating?

A

A method used to obtain a culture plate with a countable number of bacterial colonies

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39
Q

What assumption is made during dilution plating?

A

One colony comes from one bacterial cell

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40
Q

Define total viable cell count

A

A measure of the number of cells that are alive in a given volume of culture.

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41
Q

Why is dilution plating needed?

A

Because directly counting the number of colonies from a concentrated solution is difficult bc there are so many so they clump together. By dilution you can get a smaller number of cells

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42
Q

What calculation can be done to get the total viable cell count?

A

number of colonies x dilution factor

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43
Q

How can you check the accuracy of dilution plating?

A

Doing a cell count with a haemocytometer

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44
Q

What do you do if there are two or more plates with enough colonies to count in dilution plating?

A

Count the number of colonies in each plate then calculate a mean

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45
Q

How can you find the mass of microorganisms/fungi in a culture?

A

Sample of broth centrifuged/filtered to remove bacteria from liquid
Material dried until mass stabilises
Measure the dry mass overtime
Increase in mass = increase in growth

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46
Q

By what process do bacteria reproduce?

A

Binary fission

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47
Q

Define generation time

A

The time between divisions of a bacteria

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48
Q

What kind of scale is used for bacterial growth and why?

A

A logarithmic scale because the numbers are huge bc bacteria reproduce hella fast

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49
Q

What equation can be used to find the number of bacteria in a population?

A
Nt = No x 2^kt
Nt - no of organisms at time t
No - number of organisms at the start
k - exponentional growth constant
t - time
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50
Q

What equation can be used to find the exponential growth constant?

A

k=(log10Nt - log10No)/(tlog10(2))

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51
Q

How can the growth of a bacterial population be shown

A

A graph with axis time (t) against log10Nt (y)

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52
Q

Name the phases in the bacterial growth curve

A

Lag phase
Log phase
Stationary phase
Death phase

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53
Q

Explain the lag phase of the bacterial growth phase

A

Line is flat because bacteria are adjusting to their environment so growth is slow
Enzymes and genes still activating

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54
Q

Explain the log phase of the bacterial growth phase

A

Bacteria growing really fast

Growth time depends on number of nutrients and amount of space

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55
Q

Explain the stationary phase of the bacterial growth phase

A

Growth rate is zero - new cells forming = cells dying
Nutrients being used up
Waste products starting to build up

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56
Q

Explain the death phase of the bacterial growth phase

A

No. of cells dying exceeds the no of new cells forming
Nutrients have run out
Waste products have built up too high

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57
Q

Why doesn’t the exponential growth of bacteria continue forever?

A

Nutrient exhaustion - nutrient levels can’t support growth

Build up of waste products - levels can become toxic and inhibits growth (eg CO2 lowers pH)

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58
Q

How can bacteria cause infection in the body?

A

Destroying host tissue

Releasing toxins

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59
Q

How does TB cause infection?

A

Destroys host tissue

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60
Q

Name ways bacteria can enter the body

A

Wounds

Natural openings

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61
Q

Name the ways in which bacteria can be transferred

A

Droplet infection
Vector (intermediate carrier)
Direct contact
Touching the surface someone infected has touched

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62
Q

Why do bacteria produce toxins?

A

By products of their metabolism

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63
Q

Name the different types of toxins

A

Endotoxins

Exotoxins

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64
Q

What are exotoxins?

A

Soluble proteins that are produced and released into the body outside of the bacterial cell

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65
Q

What effect do exotoxins have on the body and why?

A

Widespread effect because they can travel through body fluids so they have a range of effects

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66
Q

What are endotoxins?

A

Lipopolysaccharides in the membrance of Gram negative bacteria that may be released from the bacteria if it breaks down

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67
Q

What effects do endotoxins have on the body and why?

A

Local to the site of infection because the toxins are in the bacteria for the majority of the time

68
Q

What effect does salmonella have on the body?

A

Inflammation in the gut lining

69
Q

What are antibiotics?

A

Drugs that destroy microorganisms or prevent them from reproducing

70
Q

What kind of illnesses are antibiotics used for?

A

Bacterial not viral

71
Q

What is selective toxicity?

A

Damage is done to the pathogen not the host cell

72
Q

Name the first antibiotic

A

Penicillin

73
Q

How can antibiotics be classified?

A

Bacteriocidal or bacteriostatic

74
Q

Define bacteriostatic antibiotics

A

Completely inhibit the growth of microorganisms

75
Q

Name a bacteriostatic antibiotic and its mechanism

A

Tetracyline - interferes with protein synthesis in small ribosomes in bacteria

76
Q

Define bacteriocidal antibiotics

A

Antibiotics that kill bacteria

77
Q

Name a bacteriocidal antibiotic and its mechanism

A

Penicillin breaks open bacteria cell walls

78
Q

Define broad-spectrum antibiotics

A

Destroys a wide range of harmful pathogens

79
Q

Define narrow-spectrum antibiotics

A

Targets one or two pathogens

80
Q

What does the effectiveness of a drug depend on?

A

Drug concentration
pH
Susceptibility of pathogen (is it antibiotic resistant)
Whether pathogen can destroy the antibiotic

81
Q

What is antibiotic resistance?

A

When bacteria mutate so drugs are no longer effective on them

82
Q

Why are mutations common in bacteria?

A

Because bacteria reproduce rapidly

83
Q

How does natural selection lead to antibiotic resistance?

A

Allele which causes resistance allows bacteria to survive. When bacteria reproduce, the allele is passed on quickly through other generations. Allele frequency increases.

84
Q

What is horizontal transmission?

A

When bacteria can acquire resistance from other species through the transmission of their plasmids

85
Q

How can we control the spread of antibiotic resistance?

A
  • Prescribe antibiotics sparingly
  • Complete the course of antibiotics
    -Good hygiene measures:
    Alcohol gels in hospitals
    Staff wearing clean clothing
    Isolating ill patients
    Screening patients for infection before they enter the hospital
    Monitoring levels of healthcare acquired infections
86
Q

Why are viral infections specific to particular tissues?

A

Host cells have antigens and only a few viruses have complementary receptors

87
Q

What is influenza

A

A virus which infects the tissues of the breathing system - especially the lungs

88
Q

What are the modes of transmission of influenza?

A
Droplet infection
Direct contact with:
-animal droppings
-virus filled mucus
-surfaces contaminated with the virus
89
Q

What are the modes of infection of influenza

A

Ciliated epithelial cells of the lungs are infected. Viral RNA takes over the cell, reproduces and cell releases viruses and die

90
Q

What are the pathogenic effects of influenza?

A

Headaches, runny nose, coughing, vomiting, fever etc

91
Q

How is influenxa treated?

A

No cure but antivirals used to relieve symptoms

Vaccine used as a preventative measure

92
Q

What is stem rust fungus

A

A fungus which affects cereal crops

93
Q

What are the modes of transmission of stem rust infection

A

Wind carries pores

Infected plant fragments in the soil

94
Q

What are the modes of infection of stem rust infection

A

Spores germinate in water on the plant, the fungus enters the plant through stomata and enzymes digest plant cells and nutrients are absorbed

95
Q

What are the pathogenic effects of stem rust fungus

A

Nutrients lost from plant
Stem weakened
Blisters on leaf surface
Plant loses control over water loss

96
Q

How can stem rust be controlled?

A

Dont use nitrate rich fertilisers
Use earlier maturing plants
Bigger spaces between plants
Fungicides

97
Q

What is malaria?

A

Fever caused by a parasite called Plasmodium spp.

98
Q

What is the mode of transmission of malaria?

A

Mosquito vectors transfers parasite to humans

Mouthpiece pierces the skin and the saliva has anticoagulants so blood doesnt clot while it gets its blood meal.

99
Q

What is the mode of infection of malaria?

A

Parasites travel to the liver –> RBC –> reproduce asexually then burst out of RBC —> eventually transmitted back to the mosquito taking the blood meal

100
Q

What are the pathogenic effects of malaria?

A

Shaking, fever, sweatin
Muscle pains
Liver damage
Anaemia

101
Q

Define an endemic disease

A

A disease that is constantly present in an area or country

102
Q

Given an example of an endemic disease

A

Malaria

103
Q

Why is it difficult to treat endemic diseases?

A

Widespread
Hard to get people to cooperate
No. of hosts
Expensive

104
Q

Name two ways we can control malaria

A

Prevent bites

Control number of mosquitos

105
Q

How can we prevent mosquito bites

A

Insect repellents
Mosquito nets with insecticides
Screens on door
Clothing to cover skin

106
Q

How can we control the number of mosquitos

A

Avoid standing water/sewage
Introduce predators
Pesticides
Water treatment to kill larvae

107
Q

Name some of the ethical issues with controlling diseases

A

People unsure about safety of vaccines
Difficult to obtain consent where medical education is little
Money can be spent better elsewhere

108
Q

Name some of the social issues with controlling diseases

A

People need to change habits

Change social attitudes - difficult

109
Q

Name some of the economic issues with controlling diseases

A

Treatment, control and prevention is expensive and countries are normally poor
Difficult to allocate resources

110
Q

What is the role of the scientific community in controlling diseases

A

Prevention: getting insecticide filled nets
Treatment: using drugs in combination is more effective

111
Q

What are antigens?

A

Any substance that stimulates an immune response from the body

112
Q

What substances can be antigens?

A

Proteins, glycoproteins, carbohydrates, toxins and even whole organisms

113
Q

What is a non-specific response?

A

The body’s defence against pathogens which is triggered by any pathogen

114
Q

What are the three main types of non-specific response?

A

Inflammation
Fever
Phagocytosis

115
Q

Explain the stages leading to inflammation

A

Tissue damage causes mast cells and basophils to release histamines
Blood vessels dilate - local heat and redness to reduce effectiveness of pathogens
Leaky capillaries cause leucocytes to be released and engulf pathogens and antibodies to disable them

116
Q

What are histamines?

A

Chemicals which cause blood vessels to dilate and cause capillaries to leak

117
Q

Explain why you get a fever while ill

A

Hypothalamus increases body temperature:

  • reduce the effectiveness of pathogens
  • immune system works better at a higher temperature
118
Q

What is a phagocyte?

A

A general term for an WBC that engulfs and digests pathogens any other foreign material in the blood

119
Q

Name the 2 types of phagocytes

A

Neutrophils and macrophages

120
Q

Name some characteristics of neutrophils

A

Granulocyte
Can only ingest a few pathogens before it dies
Cannot renew lysosyme stores

121
Q

Name some characteristics of macrophages

A

Agranulocyte
Made from monocytes from the tissue
Can renew their lysosyme stores

122
Q

Explain how phagocytes work

A

Engulfs pathogen
Phagosome surrounds pathogen and fuses with lysosom
Enzymes break down pathogen
Cytokines released

123
Q

What do cytokines do?

A

Signal for other WBCs

Stimulate immune response

124
Q

What are opsonins?

A

Chemicals which bind to pathogens and label them so WBCs can detect them more easily

125
Q

What is the immune response?

A

Body’s specific response to invasion by pathogens

126
Q

Name the four characteristics of the immune system

A

Distinguish self from non-self
Diverse
Specific
Immunological memory

127
Q

What are the two main types of WBC in the immune system?

A

Lymphocyte and macrophage

128
Q

What are the two types of lymphocytes in the immune system?

A

B lymphocytes

T lymphocytes

129
Q

Name the types of B lymphocytes

A

B effectors
Plasma cells
B memory

130
Q

Name the types of T lymphocytes

A

T killer
T helper
T memory

131
Q

Where do B cells get made and develop?

A

Bone marrow then develop in the lymph gland

132
Q

Where do T cells get made and develop?

A

Bone marrow then develop in the thymus gland

133
Q

What do plasma cells do?

A

Produce antibodies

134
Q

What do T killer cells do?

A

Produce chemicals to kill infected cells

135
Q

What do T helper cells do?

A

Activate plasma cells and B cell APCs

136
Q

What are the body’s two specific responses to infection?

A

The humoral response and the cell mediated response

137
Q

When is the humoral response activated?

A

When the antigens of the pathogens are outside of body cells

138
Q

Name the two main stages of the humoral response

A

T helper activation

B effector stage

139
Q

Summarise the stages of the T helper activation stage

A

Macrophage engulfs pathogen
Surrounds with phagosome and lysosome
Separates antigen and binds it to MHC
Becomes APC
T cell with complementary receptor binds to antigen
T cell divides and replicates into activated T memory and T helpers

140
Q

Summarise the stages of the effector stage

A

B cells act as macrophages and become APCs
T helper binds to B cells and releases cytokines
B cells divide into B effector and B memory
B effector –> plasma cell —> antibodies

141
Q

What is clonal selection?

A

The selection of cells that carry the right antibody for a specific antigen

142
Q

What are antibodies?

A

Glycoproteins that are released into circulation and bind to the antigen of pathogens

143
Q

How can antibodies cause pathogen deaths?

A

1) Agglutination - pathogens clump then get engulfed
2) Opsonisation - acts as an opsonin and signals for phagocytes
3) Neutralisation - neutralises toxins

144
Q

Name adaptations of plasma cells

A

Hella endoplasmic reticulum

Ribosomes

145
Q

When is the cell mediated response used?

A

When the pathogen enters the body cells

146
Q

Summarise the stages of the cell mediated response

A

Body cells acts as an APC
T killer w complementary receptor binds to APC
T helper releases cytokines so T killer rapidly divides
Release enzymes which make pores in membrane so water +ions enter membrane and cause cell to burst
Pathogens destroyed
T memory cells made

147
Q

What is the difference between the response of T memory cells and B memory cells

A

T memory cells release T killer cells

B memory cells produce antibodies

148
Q

What is the primary immune response?

A

When the body has come into contact with a pathogen and its antigen for the first time. Involves the production of antibodies and memory cells

149
Q

What is the secondary immune response?

A

When the body comes into contact with a pathogen again and memory cells are used

150
Q

What is natural active immunity?

A

When the body produces its own antibodies against a pathogen and its antigen encountered naturally

151
Q

What is natural passive immunity?

A

When antibodies are made by the mother and passed onto the baby via breastmilk or placenta

152
Q

What is immunisation?

A

The process of protecting people from infection by giving them passive or active artificial immunity

153
Q

What is a vaccination?

A

The procedure of giving someone dead or inactive pathogen to stimulate an immune response

154
Q

What is passive artificial immunity?

A

When the antibodies from one individual are extracted and injected into another

155
Q

What is active artificial immunity?

A

When the body produces its own antibodies and memory cells to an antigen from a vaccination

156
Q

How is the pathogen in a vaccination made non-infectious?

A

Toxin detoxified
Inactive/dead pathogen
Attentuated pathogens - viable but modified so they dont cause disease

157
Q

What is the only disease to be eradicated?

A

Smallpox

158
Q

Why is it not possible to totally eradicate many diseases?

A

They have multiple hosts - animals, soils and waters

159
Q

What is the elimination of a disease?

A

Where it is no longer seen in humans but exists in the environment or animal so vaccinations still needed

160
Q

What is meant by a controlled disease?

A

When a disease is still occurring but not to the extent that it is a health problem

161
Q

Define herd immunity?

A

When a large proportion of a population is immune to a pathogen (usually by vaccination) which lowers the risk of infection

162
Q

What are the pros of vaccination?

A

Person is protected against infection - dont die
Herd immunity - those who cant get immunised are still safe
Costs of treating disease minimised

163
Q

What are the cons of vaccination?

A

Some children cant be vaccinated bc of allergies or extreme immune response

164
Q

Name a bacteria which produces exotoxins

A

Staphylococcus aureus

165
Q

Name a bacteria which produces endotoxins

A

Salmonella