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Bacto practical > 1st test from plennary > Flashcards

1st test from plennary Flashcards

1
Q

What are the 2 methods when detecting bacteria in a sample

A

Direct and Indirect way

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2
Q

What are the Aims of culturing bacteria

A

• Isolation of causative agents from pathological samples
(DIAGNOSTIC AIM)

• antibiotic susceptibility testing
(for the purpose of targeted ANTIBACTERIAL THERAPY!!!)

• RESEARCH purposes

• INDUSTRY
(food industry, pharmaceutical industry, fermentation, antibiotic production etc.)

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3
Q

What do you do referring to the DIAGNOSTIC AIM

A

Isolation of causative agents from pathological samples

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4
Q

What is the purpose of targeted antibacterial therapy

A

antibiotic susceptibility testing

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5
Q

When performing Detection of bacteria in a sample the Direct way - What are the different types?

A
  1. Smear
  2. Impression Smear
  3. PCR
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6
Q

From where can you collect samples for a SMEAR when performing the direct way of bacterial sample

A
  • Nasal discharge,
  • Vaginal discharge
  • Pus from an abscess,
  • Mastitic milk,
  • Blood (septicemia),
  • Urine
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7
Q

What is impression smear? Example of a examination with an impression smear

A

Quick and simple (shape, size, staining, mixed infection

(eg. anthrax: parenchymal organs – cutted surface

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8
Q

What is the advantage of an impression smear

A

advantage: quick and simple,

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9
Q

What is the disadvantage of an impression smear

A

disadvantage: only: quantity, shape, size of bacteria

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10
Q

What is PCR and what do we use it for

A

PCR (detection of specific DNA-sequences)
– unculturable bacteria!

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11
Q

What is this

A

agar-gel electrophoresis

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12
Q
A

Smear

(sputum sample)

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13
Q
A

Smear, wound secretion

(anthrax, Bacillus anthracis)

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14
Q
A

Smear (cattle, spleen),

toluidine-blue staining

(Bacillus anthracis)

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15
Q
A

Smear from a deep anaerobic wound

(Clostridium tetani)

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16
Q
A

Smear, sheep gastrointestinal tract,

Gram-staining

(Clostridium perfringens)

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17
Q
A

Smear, mastitic milk

(Streptococcus agalactiae)

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18
Q
A

Smear, mastitic milk

(Streptococcus sp.)

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19
Q
A

Smear, human genital secretion

(Neisseria gonorrhoea)

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20
Q
A

Smear, human vaginal secretion

(Lactobacillus sp.)

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21
Q
A

Smear, urinary sediment, dog

(E. coli)

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22
Q
A

Smear, human blood

(plague, Yersinia pestis)

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23
Q

How do you perform detection of bacteria in the

Indirect way

A

- By animal trial

after death or extermination, we try to isolate the causative agent,

**- Inoculation the sample onto a medium**
we culture (isolate) bacteria on a medium, preparing pure culture, identification.
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24
Q

How can we culture Treponema pallidum?

A

INDIRECT METHODE

  • In rabit testicle ONLY
  • Treponema pallidum: human syphilis: culturable in a rabbit testicle „only”.
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25
Q

How can we culture Francisella tularensis

A

INDIRECT WAY

Francisella tularensis: tularemia, inoculation into a MOUSE

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26
Q

Composition of media

Bacteria does not need to be in harmony with the media

TRUE or FALSE

A

FALSE

The connection between bacteria and media is very closed. Must be in harmony with demand of bacteria!

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27
Q

What is the composition of Media

A

Water

C-sources

N-sources:

  • vitamins and other additives

Osmotic pressure

pH: 7.2-7.4

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28
Q

80-90% of the bacterial cell is water!

True or false

A

True

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29
Q

Which organic C-source is needed

A

GLUCOSE!!! (Pyruvic acid!)

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30
Q

Composition of media: N-source

A

N-sources:
- protein-hydrolisates: peptone (digested casein), triptone (digested muscle)

  • native protein: several species need it: eg. Trueperella pyogenes
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31
Q

Composition of media- vitamins and other additives:

B1 vitamin has a impact on which production

A
  • vitamins and other additives:
  • *- B1 vitamin:** bacteria which have impact in cheese production
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32
Q

Composition of the media, vitamins and aditives

  • B2 vitamin has an impact on:
A

most Lactobacilli

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33
Q
  • V (NAD, X-factor (haemin has a impact on which species?
A
  • V (NAD, X-factor (haemin: Glaesserella, Avibacterium, Actinobacillus spp.
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34
Q
  • Brucella: Which components are needed
A

- Brucella: B1 vitamin, nicotinic-acid-amide, pantothenic-acid is needed

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35
Q
  • Erysipelothrix rhusiopathiae: Which components are needed
A

- Erysipelothrix rhusiopathiae: para-amino-benzoic-acid (PABA) is needed

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36
Q

Composition of media

OSMOTIC PRESSURE

A

In a par with 0.85% NaCl solution

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37
Q

COmposition of media

PH

A

- pH: 7.2-7.4

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38
Q

Classification (grouping) of media

According to

A
  1. ORIGIN
  2. STATE
  3. AIM of CULTURE
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39
Q

Classification (grouping) of media

According to • Origin

A

1- natural (potato slide, blood, serum, milk, bile, urine)

2- artificial

3- synthetic (=chemically defined) - the exact amount of ingredients are known

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40
Q

Classification (grouping) of media

According to

State

A

(liquid, semisolid, solid)

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41
Q

Classification (grouping) of media

According to

Aim of culture

A

1- common (=basic nutrient) - these are capable of sustaining growth of the less fastidious bacteria

2- selective - contain inhibitory substances that prevent the growth of unwanted bacterial species

3- differential (=indicator) - They are designed to give a presumptive identification of bacterial colonies due to the biochemical reactions in the media. They often contain a fermentable sugar plus a pH indicator that gives a colour change in the media.

42
Q

Bacterial growt show what in a liquid samlpe

A

TURBIDITY

Sterile are clear

43
Q

Which bacteria

A

Mycobacterium phlei

44
Q

Name arrows from top to bottom

A
  1. Pellicle
  2. No turbidity
  3. Sediment
45
Q

Which bacteria

A

Staphylococcus aureus

46
Q

Name bacteria

A

Bacillus subtilis

47
Q

Name bacteria

A

Escherichia coli

48
Q

Which medium is made from Sea weed

A

Agar - Agar

49
Q

Which bacteria can be cultured in agar agar

A

Gelidium sp., Gracilaria sp.

50
Q

Name a special characteristic of the agar agar powder mediun

A

DIFFERENT MELTINGPOINT and FREEZING Point

Agar-agar powder

51
Q

DIFFERENT MELTINGPOINT and FREEZING Point

in agar agar powder, but what is is

A

melting-point:

85-90 °C

solidifying-point:

45-50 °C

52
Q

Semi solid test are used for

A

Motillity test

53
Q

Which media

Which test

Which is positive and negative

A

SEMI SOLID MEDIUM

MOTILLITY TEST

LEFT is NEGATIVE and RIGHT is POSITIVE

54
Q

Name of test in which medium

A

Oxidation/Fermentation test

– semisolid medium!

(Do NOT mix with oxidase test!!!)

55
Q

Left medium

A

SOLID MEDIUM

Blood agar

10% sheep or ox blood

Solid, artificial, differential medium

Solid Medium

56
Q

Right medium

A

Solid, Artificial, Common medium

Nutrient agar

57
Q
A

Non-haemolytic colonies (Pasteurella multocida)

58
Q
A

Haemolysis

Streptococcus pneumoniae

alpha (partial, incomplete, biliverdin)

59
Q
A

Hemolysis

Streptococcus equi

Beta (compleete)

60
Q
A

Carbohydrate broths

Mannitol, Dextrose, Lactose, Sucrose

Liquid, artificial, differential medium

(Phenol red – pH indicator)

UNINOCULATED - NEGATIVE - POSITIVE

61
Q

Inhibitory materials:

pH indicator:

A

MAC CONKEYS AGAR

Solid Artificial Selective Differential medium

Lactose neg - inoculated - positive

Inhibitory materials:

  • bile salts crystal violet

pH indicator:

  • neutral red
62
Q
A
63
Q
A
64
Q
A
65
Q

PREPARATION of MEDIA

A

Dehydrated media

  • wide range
  • easy to use
  • dissolved in bidest. water, boiled - Sterilisation (autoclaving!)
66
Q
A

AUTOCLAVE!!! (121degreed, 15 minutes)

67
Q
A

Plastic Petri-dishes

  • Gamma-steril (irradiated)
  • 90 mm in diameter
  • 20 ml medium
68
Q
A

BLOOD AGAR

10% defibrinated sheep or ox blood

Add 10% blood
to the cooled medium (45-50 °

69
Q
A

Chocolate agar

Heat treatment:

8 0 ° C , in 20 m i n

70
Q

Bacterias suitable for chocolate agar

A

Avibacterium paragallinarum

Actinobacillus pleuropneumoniae

Glaesserella (Haemophilus) parasuis

Taylorella equigenitalis

Histophilus somni

71
Q
A

Inoculating loop!!!

72
Q
A

Inocculating loop and bunsen burner

73
Q
A

Fowl typhoid

(Salmonella enterica
(st. Gallinarum))

74
Q
A
75
Q
A

Inoculation of a sample onto the surface of a solid medium

  • *- specimen** (liver, spleen, lung, pus, mastitic milk, urine)
  • *- Petri dish** is first labelled, on the agar side

(type of specimen, date of inoculation - waterproof marker pen)

  • burn the surface of the organ with a hot plate shaped metal instrument (e.g. with an old scalpel), decontamination
  • stab with the inoculating loop in the organ through the burned surface
  • pull out the inoculating loop from the organ, and streak the agar plates
76
Q
A

Laminar flow box (biosafety cabinet)

77
Q
A

Incubator (37 °C)

78
Q

What kind of culture

A

PURE CULTURE!!!

  • descendants of the same bacterium cell
  • same genotype
  • same phenotype (cultural, morphological, biochemical characteristics)
79
Q

What heppend here

A

Airborn contamination

80
Q

What are the circumstances of incubation

A
  • atmosphere
  • temperature
  • time of incubation
81
Q

What does Obligate aerobe bacteria mean

A

They can grow in the presence of O2 only.

82
Q
A
83
Q

Name Obligate GRAM + bacterias

A
  • Bacillus
  • Dermatophilus • Micrococcus
  • Nocardia
  • Rhodococcus
84
Q

Name Obligate GRAM - bacterias

A
  • Brucella
  • Bordetella bronchiseptica • Francisella
  • Moraxella
  • Pseudomonas
85
Q

Staining of Obligate bacterias

MYCOBACTERIUM

A

ZIEHL- NEELSEN staing

86
Q
A
87
Q

What does obligate anaerobe bacteria mean

A

They can not tolerate O2 at all.

88
Q

Name Obligate anaerobe Gram + bacteria

A
  • Actinomycesbovis
  • Peptostreptococcus

• Clostridium

89
Q

Name obligate gram - anaerobe bacterias

A
  • Bacteroides
  • Dichelobacter
  • Eubacterium
  • Fusobacterium
  • Brachyspira
  • Campylobactermucosalis
90
Q

What does

Aerobe, facultative anaerobe bacteria mean

A

They can grow in the presence or in the abscense of O2, too.

91
Q

What are the facultative anaerobe GRAM + bacteria

A

Corynebacterium

Erysipelothrix

rhusiopathiae

Listeria

Staphylococcus

Streptococcus

92
Q

What are the facultative anaerobe GRAM - bacteria

A

• Actinobacillus
• Aeromonas
• Enterobacterium

• Haemophilus
• Pasteurella
• Vibrio

93
Q

What is

Microaerophile bacteria

A

4-6% partial O2 pressure is needed

94
Q

Microaerophile GRAM + and GRAM -bacteria

A

GRAM +

  • several Actinomycessp.
  • Lactobacillus

GRAM -

• Campylobacter spp.

95
Q

What is

Capnophile bacteria

A

They need 5-10% CO2

96
Q

Caphnophile GRAM - bacterias

A
97
Q

Caphnopile GRAM + bacterias

A

Gram+

  • Actinomycesviscosus
  • Dermatophilus congolensis
98
Q

What does the picture show

A

Where the bacteria grow

  1. AEROBE
  2. Microaerophile
  3. Faculative Anaerobe
  4. Anaerobe
99
Q

Anaerobic cultures

A

Anaerobic culture

  • candle jar method
  • chemical
  • pirogallic acid + KOH
  • H2 + palladium catalisator
  • Na-borohidrid + NaHCO3 + ascorbic acid + H2O •other

•evacuation of air with a vacuum pump

  • anaerobic broth
  • deep semisolid agar
  • pre-reduced media
100
Q
A

Light microsope

visible light (400-700 nm) immersion objective and imm. oil resolving power: 0.2-0.4 μm magnification: 1200-1500×

101
Q

How is this image taken

A

Darkfield microscope

Special condenser, illuminate with oblique ray of light, dark background, corpuscular elements (bacteria) are glittering,

examination of bacterial motility

102
Q

What do we examine with dark field microscope

A

examination of bacterial motility

Bacto practical (9 decks)

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