2. Identification of bacterial cells Flashcards

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1
Q

name two culture collections across the world

A

American type culture collection (ATCC)

national collection of type cultures (NCTC) -> public health england

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2
Q

what does NCIMB stand for

A

national collection of industrial, food and marine bacteria

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3
Q

how many classes of bacteria are there

A

4

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4
Q
which class of bacteria:
1. can cause severe disease?
A

class 3

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5
Q
which class of bacteria:
2. is safe to use (non-pathogenic)
A

class 1

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6
Q
which class of bacteria:
3. is incurable
A

class 4

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7
Q

how many strains of class 4 bacteria are there

A

none

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8
Q

name a class 4 virus

A

ebola

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9
Q

what are class 2 bacteria

A

cause mild, treatable disease, can be worked with in the lab

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10
Q

how does Gram staining work

A

crystal violet applied to samples, if Gram-positive, crystal violet will bind tightly to peptidoglycan and following an ethanol wash, be stained purple. (positively purple)

if not, following ethanol wash, and counter staining with safranin, will stain pink
(negative pink)

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11
Q

we can identify shapes based on morphology, name some shapes

A

cocci (circle)
rods
spiral
corkscrew

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12
Q

how do we measure oxygen tolerance

A

inoculate broths with bacterium, note location of growth

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13
Q

where will obligate aerobe’s grow in broth medium, what does this show?

A

at the top

- preference for oxygen

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14
Q

where will obligate anaerobe’s grow in broth medium, what does this show?

A

growth at the bottom

- no tolerance for oxygen

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15
Q

where will aerotolerant anaerobe grow in broth medium, what does this show?

A

bacterium distributed throughout, tolerance for presence and absence of oxygen

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16
Q

how can we identify the presence of endospores

A

malachite green stains bacteria black and endospores appear white

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17
Q

what type of cells are endospores found in

A

Gram positive bacteria only

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18
Q

where do endospores form under, what does this mean?

A

unfavourable conditions

they are highly resilient to heat, drying, radiation and chemicals

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19
Q

how do you remove endospores

A

a strict course of antibiotics taken for months to eradicate further infections

20
Q

describe the structure of endospores (2)

A
  1. contain calcium dipicolinate

2. have special proteins to protect their DNA.

21
Q

how can cell motility be measured

A

place a sterile needle containing the bacterium colony into sloppy agar. this medium contains a dye which stains cells red.

non-motile bacterium will remain in one place, motile bacterium will move around the medium.

22
Q

what does catalase convert

A

toxic hydrogen peroxide into water and oxygen

23
Q

how do you perform the catalase test, what would a positive result show?

A
  • adding a bacterial culture to a bubble of hydrogen peroxide
  • if catalase is present, bubbles of oxygen will be produced
24
Q

what does kovac’s reagent test for

A

the presence of cytochrome C and oxidase

25
Q

what is a positive oxidase test result

A

change in colouration from white to purple

26
Q

what can a melt curve show to help us classify bacteria

A
  • melting temperature is relative to nucleotide composition

- higher guanine-cytosine concentration = higher melting temperature = more energy required to separate the strands

27
Q

how can we test whether a bacterium uses lactose

A

placing bacterium in a growth medium containing only lactose as the sugar source , we can then measure growth

28
Q

how can we detect growth in lactose medium

A

when there is growth, the pH changes and oxygen is produced, the solution turns from red to yellow

29
Q

what infection is known to produce black insoluble hydrogen sulphide

A

salmonella

30
Q

what is hydrogen sulphide produced from

A

fe2+ and S2-

31
Q

how do media kits work for identifying clinical isolates

A

test kit contains various dehydrated tests
you add the bacterium to each test strip and incubate for 24 hrs

then measure colour change
each change is assigned a number value which using the website helps inform what microbe you have

32
Q

how do macConkey agar plates help to distinguish bacteria

A

contains a pH indicator,
lactose fermenting bacteria produce pink colonies,
non-lactose fermenting bacteria will produce yellow colonies

33
Q

how do chromagar plates help to distinguish bacteria

A

medium contains artificial substrates that when hydrolysed by yeast produce specific colourations

e.g. candida tropicalis = blue

34
Q

what do beta haemolysis zones show, what bacteria does this

A

show where RBCs have been digested,

e.g. streptococcus

35
Q

what medium enhances N. gonorrhoeae growth

A

thaymer-martin medium

36
Q

Name 2 selective media

A

Thaymer-Martin (gonorrhoea)

Mac Conkey agar

37
Q

How is macconkey agar selective

A

Contains bile salts = prevents Gram-positive growth

38
Q

Name two differential media

A

Blood agar

Chromagenic

39
Q

Chromagar is both a nutrient and differential media, why?

A

Funghal produce enzymes that react with different compounds in the medium, producing colonies of different colours

40
Q

What is Ziehl- Neesen another name for

A

Acid fast technique

41
Q

How does ziehl-neesen work

A

Uses fluorescence or stain to highlight microorganisms

42
Q

What is Ziehl- neesen used for

A

Cryptosporidium

43
Q

What’s is an advantage of Ziehl- neesen

A

Rapid results, cheap, targeted to the pathogen

44
Q

Give an example of a coloured colony produced on chromager

A

Candida tropicalis = blue colonies

45
Q

Describe chocolate agar, give an example of the colonies produced

A

Differential media - Identification of colony colour
Contains lysed RBCs

Neisseria gonorrhoea = white colonies
Haemophililus = colourless

46
Q

How is blood agar differential

A

3 different types of haemolysis can be seen on this plate

- alpha, beta, gamma (no haemolysis)

47
Q

Give an example of a pathogen thAt conducts beta haemolysis

A

Streptococcus pyogenes