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Bacto practical > 1st test from plennary > Flashcards

1st test from plennary Flashcards

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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
How can we culture Francisella tularensis
INDIRECT WAY Francisella tularensis: tularemia, inoculation into a MOUSE
26
Composition of media Bacteria does not need to be in harmony with the media TRUE or FALSE
FALSE The connection between bacteria and media is very closed. Must be in harmony with demand of bacteria!
27
What is the composition of Media
Water C-sources N-sources: - vitamins and other additives Osmotic pressure pH: 7.2-7.4
28
80-90% of the bacterial cell is water! True or false
True
29
Which organic C-source is needed
GLUCOSE!!! (Pyruvic acid!)
30
Composition of media: N-source
N-sources: - protein-hydrolisates: **peptone** (digested casein), **triptone** (digested muscle) - **native protein**: several species need it: eg. **Trueperella pyogenes**
31
Composition of media- vitamins and other additives: ## Footnote **B1 vitamin has a impact on which production**
- vitamins and other additives: * *- B1 vitamin:** bacteria which have impact in **cheese production**
32
Composition of the media, vitamins and aditives - **B2 vitamin** has an impact on:
**most Lactobacilli**
33
- **V (NAD, X-factor (haemin has a impact on which species?**
- **V (NAD, X-factor (haemin**: Glaesserella, Avibacterium, Actinobacillus spp.
34
- Brucella: Which components are needed
**- Brucella: B1 vitamin, nicotinic-acid-amide, pantothenic-acid is needed**
35
- Erysipelothrix rhusiopathiae: Which components are needed
**- Erysipelothrix rhusiopathiae:** para-amino-benzoic-acid (**PABA**) is needed
36
Composition of media OSMOTIC PRESSURE
In a par with **0.85% NaCl solution**
37
COmposition of media PH
**- pH: 7.2-7.4**
38
Classification (grouping) of media According to
1. ORIGIN 2. STATE 3. AIM of CULTURE
39
Classification (grouping) of media According to • Origin
1- natural (potato slide, blood, serum, milk, bile, urine) 2- artificial 3- synthetic (=chemically defined) - the exact amount of ingredients are known
40
Classification (grouping) of media According to State
**(liquid, semisolid, solid)**
41
Classification (grouping) of media According to **Aim of culture**
**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
Bacterial growt show what in a liquid samlpe
TURBIDITY Sterile are clear
43
Which bacteria
Mycobacterium phlei
44
Name arrows from top to bottom
1. Pellicle 2. No turbidity 3. Sediment
45
Which bacteria
Staphylococcus aureus
46
Name bacteria
Bacillus subtilis
47
Name bacteria
Escherichia coli
48
Which medium is made from Sea weed
Agar - Agar
49
Which bacteria can be cultured in agar agar
Gelidium sp., Gracilaria sp.
50
Name a special characteristic of the agar agar powder mediun
DIFFERENT MELTINGPOINT and FREEZING Point ## Footnote Agar-agar powder
51
DIFFERENT MELTINGPOINT and FREEZING Point in agar agar powder, but what is is
melting-point: 85-90 °C solidifying-point: 45-50 °C
52
Semi solid test are used for
Motillity test
53
Which media Which test Which is positive and negative
SEMI SOLID MEDIUM MOTILLITY TEST LEFT is NEGATIVE and RIGHT is POSITIVE
54
Name of test in which medium
Oxidation/Fermentation test – semisolid medium! (Do NOT mix with oxidase test!!!)
55
Left medium
SOLID MEDIUM Blood agar 10% sheep or ox blood Solid, artificial, differential medium Solid Medium
56
Right medium
Solid, Artificial, Common medium Nutrient agar
57
Non-haemolytic colonies (Pasteurella multocida)
58
Haemolysis ## Footnote Streptococcus pneumoniae alpha (partial, incomplete, biliverdin)
59
Hemolysis Streptococcus equi Beta (compleete)
60
Carbohydrate broths Mannitol, Dextrose, Lactose, Sucrose Liquid, artificial, differential medium (Phenol red – pH indicator) UNINOCULATED - NEGATIVE - POSITIVE
61
Inhibitory materials: pH indicator:
MAC CONKEYS AGAR ## Footnote Solid Artificial Selective Differential medium Lactose neg - inoculated - positive Inhibitory materials: - bile salts crystal violet pH indicator: - neutral red
62
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65
PREPARATION of MEDIA
Dehydrated media - wide range - easy to use - dissolved in bidest. water, boiled - Sterilisation (autoclaving!)
66
AUTOCLAVE!!! **(121degreed, 15 minutes)**
67
Plastic Petri-dishes - Gamma-steril (irradiated) - 90 mm in diameter - 20 ml medium
68
BLOOD AGAR **10% defibrinated sheep or ox blood** **Add 10% blood** to the c**ooled medium (45-50 °**
69
Chocolate agar Heat treatment: **8 0 ° C , in 20 m i n**
70
Bacterias suitable for chocolate agar
Avibacterium paragallinarum Actinobacillus pleuropneumoniae Glaesserella (Haemophilus) parasuis Taylorella equigenitalis Histophilus somni
71
Inoculating loop!!!
72
Inocculating loop and bunsen burner
73
Fowl typhoid (Salmonella enterica (st. Gallinarum))
74
75
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
Laminar flow box (biosafety cabinet)
77
Incubator (37 °C)
78
What kind of culture
PURE CULTURE!!! * descendants of the same bacterium cell * same genotype * same phenotype (cultural, morphological, biochemical characteristics)
79
What heppend here
Airborn contamination
80
What are the circumstances of incubation
- atmosphere - temperature - time of incubation
81
What does Obligate aerobe bacteria mean
They can grow in the presence of O2 only.
82
83
Name Obligate GRAM + bacterias
* Bacillus * Dermatophilus • Micrococcus * Nocardia * Rhodococcus
84
Name Obligate GRAM - bacterias
* Brucella * Bordetella bronchiseptica • Francisella * Moraxella * Pseudomonas
85
Staining of Obligate bacterias MYCOBACTERIUM
ZIEHL- NEELSEN staing
86
87
What does obligate anaerobe bacteria mean
They can not tolerate O2 at all.
88
Name Obligate anaerobe Gram + bacteria
* Actinomycesbovis * Peptostreptococcus **• Clostridium**
89
Name obligate gram - anaerobe bacterias
* Bacteroides * Dichelobacter * Eubacterium * Fusobacterium * Brachyspira * Campylobactermucosalis
90
What does ## Footnote Aerobe, facultative anaerobe bacteria mean
They can grow in the presence or in the abscense of O2, too.
91
What are the facultative anaerobe GRAM + bacteria
Corynebacterium Erysipelothrix rhusiopathiae Listeria **Staphylococcus** **Streptococcus**
92
What are the facultative anaerobe GRAM - bacteria
• Actinobacillus • Aeromonas **• Enterobacterium** • Haemophilus **• Pasteurella** • Vibrio
93
What is ## Footnote Microaerophile bacteria
4-6% partial O2 pressure is needed
94
Microaerophile GRAM + and GRAM -bacteria
GRAM + * several Actinomycessp. * Lactobacillus GRAM - • Campylobacter spp.
95
What is ## Footnote Capnophile bacteria
They need 5-10% CO2
96
Caphnophile GRAM - bacterias
97
Caphnopile GRAM + bacterias
Gram+ * Actinomycesviscosus * Dermatophilus congolensis
98
What does the picture show
Where the bacteria grow 1. AEROBE 2. Microaerophile 3. Faculative Anaerobe 4. Anaerobe
99
Anaerobic cultures
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
Light microsope visible light **(400-700 nm)** immersion objective and **imm. oil r**esolving power: 0.2-0.4 μm magnification: **1200-1500×**
101
How is this image taken
Darkfield microscope **Special condenser,** illuminate with **oblique ray of light**, dark background, **corpuscular elements (bacteria) are glittering,** examination of **bacterial motility**
102
What do we examine with dark field microscope
examination of **bacterial motility**

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