BACTE STAINING-CONTROLS Flashcards

1
Q
  • Directed towards coloring the cells’ shape/forms.
A

simple staining

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2
Q

example of simple staining

A

methylene blue

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3
Q

— Divide bacteria → separate groups

A

differential staining

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4
Q

— Directed towards coloring the elements’ components.

A

differential staining

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5
Q

examples of differential staining

A

gram staining
AFB staining

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6
Q

differential staining steps

A

primary stain > mordant > decolorizing agnet > secondary stain/counterstain

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7
Q

GRAM STAIN
— #____________
— Primary stain: __________
— Secondary stain: __________
— Mordant: __________
— Decolorizing agent:__________

A

differential stain

crystal violet
safranin
iodine
acetone OH

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8
Q

cell effects of crystal violet

A

purple or blue

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9
Q

cell effects of iodine

A

remain purple/blue

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10
Q

cell effects of acetic OH

A

gram +: purple or blue
gram -: colorless

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11
Q

cell effects of safranin:

A

gram +: purple or blue
gram -: pink or red

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12
Q

all cocci= gram [+/-]
except__________ [+specific 2:]

A

gram [+]
neisseria
veilonella
branhamella

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13
Q

all bacilli= gram [+/-]
except [1O]

A

arcanobacterium
bacillus
clostridium
corynebacterium
listeria
mycobaterium
nocardia
streptomyces
erysipelothrix
trophyrema whipplei

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14
Q

— Anionic dyes with ANODE groups that adhere → STAINS to CATHODE molecules.

A

Acidic dyes

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15
Q

acidic dyes example

A

acid fuchsin
eosin

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16
Q

— Cationic dyes with CATHODE groups that adhere → STAINS to ANODE molecules [nucleic acids, proteins].

A

basic dyes

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17
Q

basic dyes

A

methylene blue
malachite green
crystal violet
safranin

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18
Q

WHY GRAM-POSITIVE BECOME GRAM-NEGATIVE BACTERIA?
[main reason]

A

Heads-up

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19
Q

HEADS-UP
— Removal of ______ [X]
* ______cells- these cells may lose their
ability to [X]retain strains
— Antibiotic-treated bacterial
cells have [Reaction]
— Using ______________ [during staining]
— Due to a _________ error or the wrong use of __________

A

MgRNA
Old/Autolyzing/Dying cells
Atypical stain rxn
acidic iodine
technical
stains

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20
Q

3 EXCEPTION IN GRAM STAINING
Organisms that _______________
[Bacteria]

Organisms that _____________
[2 Bacterias]

Organisms with __________ to be resolved by __________
[Bacteria]

A

Chlamydia

mycoplasma, Ureaplasma

insufficient dimension
LM
Spirochetes

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21
Q

—- Used to stain bacteria that have high lipid contents in their cell wall

A

ACID-FAST STAINING

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22
Q

Primary stain: __________
Secondary stain: __________
— Mordant: _________ [Method]
— Tergitol: _________ [Method]

A

carbol fuchsin
methylene blue/malachite green
heat [Ziehl-Neelsen method]
cold [KInyoun method]

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23
Q

— Cell wall of _____ resists the acid- OH in decolorizing step.

A

AFB

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24
Q

ACID-FAST STAINING METHOD
1. [Method]
2. [Method]
3. [Method] - differentiate [2 Bacterias]
4. [Method] - differentiate [2 Bacterias]
5. [Method] - selective for the cell wall of ______.

A

heat/Ziehl-Neelsen Method
cold/Kinyoun
Pappenheim
Baumgarten
Auramine-rhodamine

AFB

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25
Q

MODIFIED ACID-FAST STAINING METHOD

A

Modified Kinyoun

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26
Q

— ID of intestinal coccidian oocysts

A

Modified Kinyoun

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27
Q

— Ideal for cryptosporidia and cyclospora parasites

A

Modified Kinyoun

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28
Q

MODIFIED KINYOUN
— Specimen : ______
— Rgt: = __________

A

stool
conventional acid-fast

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29
Q

MODIFIED KINYOUN
Result: ______ appear as [color]-stained organisms against a [color] background

A

oocysts
magenta
blue

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30
Q

If the result is:

Bacteria: [Method]
Parasite: [Method]

A

Acid-fast/Kinyoun
Modified acid-fast/Modified kinyoun

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31
Q

Staining technique of the ff:
Cell wall
Capsule

A

Dyar
Anthony’s. Hiss and Gin’s +Nigrosin

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32
Q

Staining technique of the ff:
Metachromatic Granules
Endospore

A

Neisser + Albert
Domer+ Schaeffer-Fulton

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33
Q

Staining technique of the ff:
Flagella
Spirochetes

A

Gray + Leifson
Levaditi’s + Fontana-Tribondeau

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34
Q

Staining technique of the DNA:

A

Feulgen

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35
Q

process that [X] Defined scope of microorganisms including some spores

A

Disinfection

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36
Q

Disinfectant can be either?

A

Physical
Chemical

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37
Q

[X] pathogenic bacteria so items are safe to handle or dispose of

A

Decontamination

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38
Q

WHY THE BACTERIA WON’T DIE?

A

organism type,number
disinfecting agent conc
nature of surface to be disinfected
presence of organic material
contact time
temp
pH
biofilms
compatibility of disinfectant <> sterilant

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39
Q

[+]Spores

A

Calcium, Protein, Carbs, Lipids, Dipicolinic acid

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40
Q

Opposite for virus with lipid-rich envelope detergent + wetting agent.

A

Mycolic acid in bacteria

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41
Q

nonliving; abnormal infectious proteins

A

Prions

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42
Q

Prions resistant to: [3]

A

heat 121C
radiation
chemicals [acid/base]

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43
Q
  • determines the exposure time that is necessary for 99.9% elimination
A

Microbial load

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44
Q

–- Preparation, dilution and use must be followed carefully by this
[2 examples]

A

Disinfecting agent conc.
● 70% alcohol
● Povidone-iodine

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45
Q

NO disinfecting agent, full contact= presence of ________

A

organic material

46
Q

__________________
Nature of the equipment may affect, so it is important to consider whether the material need to be ONLY: ________ or _________?

A

Nature of surface to be disinfected

disinfected
sterilized

47
Q

CONTACT TIME
–- Identify the ________ present first
–- Contact time may ONLY ______ / _______

Betadine - ______ minutes

A

Mx
disinfect/sterilize

1-2 mins

48
Q

Disinfectant:_________°C.
● ____°F (____°C) = Killing activity

A

2O-22C

49F/ 65C

49
Q

PH
–- ______ of bacteria desired to be killed
–- _______

A

ID
Surface ID

50
Q

BIOFILMS
–- May need to [INC/DEC] CONC of _______ / _______ time

A

INC

Disinfectant
Contact time

51
Q

COMPATIBILITY OF DISINFECTANT AND STERILANT
— 2 disinfectants [ARE/ ARE NOT] better than 1
–- __________ may negate other disinfectant.
● Example

A

ARE NOT

Disinfectants

bleach & quarternary compounf

52
Q

PHYSICAL
Sterilization [5]

A

Incineration
Moist Heat
Dry Heat
Filtration
Ionizing Radiation

53
Q

PHYSICAL
DIsinfection [3]

A

Boiling
Pasteuring
Non-Ionizing radiation

54
Q

CHEMICAL
Sterilization [2]

A

Ethylene Oxide
Hydrogen Peroxide

55
Q

CHEMICAL
Sterilization/Disinfection [4]

A

Alcohol
Antiseptics
Acetic Acid
Phenolics
Hypochlorite
Quarternary Ammonium Compound

56
Q

INCINERATION
–- Burned to ____ [____-____]
— Good for ______
— ______ method
CONS:
[2]

A

87O-98OC
prions
SAFEST

toxic air emissions
[+] heavy metals

57
Q

DRY HEAT
— [Shorter/Longer] exposure time
— Done for ____ hours
— [TEMP]
Used for [cold/heat-stable] substance such as [2]

A

Longer
1.5-3hrs
16O-18OC
heat: porcelain/glassware

58
Q

MOIST HEAT [HEAT UNDER STEAM PRESSURE]
–- [#] atm / [#] pounds per sq in. [TEMP] [#] minutes
— [Slower/Faster] than dry heat
–- Moist heat must come in contact with ______.
- [Bacteria example]
- Biological Indicator: checks if it is sterilized on the inside

A

1 atm/ 15 pounds
121C
4
Faster
surface

Bacillus stearothermophilus
Biological indicator

59
Q

MOIST HEAT
●Culture Media:
- ____ °C= 30 mins
- ____ °C= 4 mins
● Biological waste:
- 132 °C= ______

A

121
132

30 - 60 mins

60
Q

Moist heat [can also/ cannot] be used for prions

A

Can also

61
Q

Steps in Moist Heat [6]

A
  1. Autoclave [134C: 18mins, 132C: 1hr]
  2. Immerse in 1n NaOH for 1hr
  3. Rinse
  4. Autoclave [134C/121C: 1hr]
  5. Repeat Step 2-4[121C]
  6. Clean routinely
62
Q

Liquid Filtration
- uses [3]

A

cellulose acetate
nitrate membrane
vacuum

63
Q

Filtration Of Bacteria,Yeast,Molds
- Uses _____um pores of membrane filter
- Range:
- [X] # bacteria/fungi but NOT
_____

A

0.45um

0.2-0.45um

virus

64
Q

Uses 0.22um membrane filters for parenteral solutions + alcohol

A

critical sterilizing

65
Q

[X] vegetation cells + spores but NOT virus

A

critical sterilizing

66
Q

uses HEPA filter
- [X] >0.3um sized-Mx

A

Air filtration

67
Q

under moist heat, it is the fastest and simplest, all organisms + spores [except for prions]

A

autoclaving

68
Q

Autoclaving is operated by a laboratory technician. Before opening, make sure that the pressure or PSI is completely_____.

A

0

69
Q

principle is steam under pressure

A

Autoclaving

70
Q

moist heat/autoclaving biological indicator

A

Bacillus stearothermophilus

71
Q

Autoclaving for media, liquids, pipettes, utensils, and etc.
[Temp]
[psi]
[time]

A

121C 15 psi= 15-3Omins
132C= 4mins

72
Q

Autoclaving for biological wastes
[Temp]
[psi]
[time]

A

132C 15psi= 3O-6Omins

73
Q

Indication of the ff in Bacillus stearothermophilus:

Black Masking tape
Black Stripes

A

exposed to extreme convention

sterilized

74
Q

— made up of fibrous or granular materials
Example [3]

A

Depth Filters

Berkefield/Chamberland filter
Abestos

75
Q

__________filter for Porous membranes [0.1 um thick]
[1 example]

A

membrane filters

filter paper

76
Q

cellulose acetate, polycarbonate

A

membrane filters

77
Q
  • sterilize pharmaceuticals, ophthalmic solution, culture media, antibiotics and oil products
A

membrane filters

78
Q

____________
* bacteriostatic
* [-] metabolism rate
* Important in food microbiology
* Blood products

A

Low/Cold Temperature

79
Q

__C -__C for ___ hours – kills syphilis

A

2
8

72

80
Q

disruption of metabolism involves removing of water from microbes.

A

Dessication

81
Q

protein changes + DEC. chemical reaction.
* Powdered form

A

Lyophilization

82
Q

RADIATION-IONIZING
–- [Hot/Cold] Sterilization
— Causes ____ mutation +_______ prod.
— destroys [2]
Biological Indicator:
- __________

A

Cold

DNA
Peroxidase

vegetative
endospores of both prokaryotes & eukaryotes

Bacillus pumilus

83
Q

Radiation Ionizing
— Gamma rays [1500 to_______ radiation] & Xrays

A

2500

84
Q

Best for:
- gloves, plastic syringe, catheters

A

Radiation-Ionizing

85
Q

BOILING
— ____ °C for ___ mins.
— kills only _______ bacteria [sporulating/non-sporulating].
* [Example]

A

1OOC
15mins

vegetative
non-sporulating

Water bath

86
Q

PASTEURIZATION
— ____°C for ____ mins.
— kills ______ pathogens +Mx W/O DEC. ______ nutrition or _____ for _______.
— Sterilize [3]
— CANNOT eliminate bacterial _________
* [Partial/Total] Sterilization

A

7OC 3Omins
food
food
flavor
spoilage

milk, dairy products, OH

endospores

87
Q

TYNDALIZATION
—_____°C for ____ minutes for ___ consecutive days
— kills [2]
— __________ Sterilization.
* Example: _______ sterilizer [free-flowing steam]

A

1OOC 3O mins

vegetative, endospores

Fractional/Intermittent

Arnold’s

88
Q

NON-IONIZING RADIATION
— LONG ________ LOW ______
— [penetrate/do not penetrate] well
—- Damage to cellular ____ by producing _______.

A

wavelength
energy

do not penetrate

DNA
thymine dimers

89
Q

— Used on:

—- exposed surface + operating rooms

—- gloves, plastic syringe, catheters

A

Non-Ionizing Radiation

Ionizing Radiation

90
Q

— # chemical sterilant

A

Ethylene oxide

91
Q

Used for sterilizing heat-sensitive objects

A

Ethylene Oxide

92
Q

CONS: lengthy cycle time + potential health hazardq

A

Ethylene Oxide

93
Q

–-used if stabilized w/ phytate, citrate or malonate.

A

Hydrogen peroxide

94
Q

This form of Hydrogen peroxide is used to sterilize HEPA filters

A

Vapor form

95
Q

It has NO toxic by-products

A

Hydrogen peroxide

96
Q

–- Non-sporicidal and evaporates quickly.

A

Alcohols

97
Q

— As antiseptic/ thermometer /injection vial rubber septa :limited to the skin

A

Alcohols

98
Q

Alcohols’ Water content– hydrolyzes ______ in bacterial protein

A

H bonds

99
Q

— ALL Mx-cidal + sporicidal

A

Hypochlorite

100
Q

HYPOCHLORITE
— _____ ratio [bleach to water]
— Inexpensive yet ______.
— [X] DEC. capacity based on quality of ______.

CONS:
–[2] [3 physiologic] +
–addition with [2] = toxic gas

A

1:1O
effective
water

corrodes metal
discolors fabrics

oropharyngeal irritation
esophageal irritation
ocular irritation

ammonia/acids

101
Q

ALDEHYDE
— [X] [3]
–- __________Technique
Example: _________

A

corrode lenses
rubber
metals

Cold Sterilization Technique

Glutaraldehyde

102
Q

2 CHEMICAL METHODS STERILIZATION / DISINFECTION that use Cold Sterilization Technique

A

Aldehyde
Periacetic Acid

103
Q

it is best used if the equipment has organic material

A

Periacetic acid

104
Q

HEAVY METALS
— [X] microorganisms by ________________.
-– [5 metals examples]

— __________: eye drop for Neisseria gonorrhoeae.
— __________: antiseptic.

A

inactivating/precipitating cell proteins

Copper
Arsenic
Mercury
Silver
Zinc

AgNO3/Silver Nitrite
Mercuric chloride

105
Q

— 1ST widely used antiseptic/
disinfectant

A

Phenols

106
Q

PHENOLS
– ______-% Phenol
- ____ to ____ mins contact
time
–- Effective against ________
— [X] [2]

A

5
1O-3O mins

Mycobacteria

plasma membrane
protein

107
Q

IODINE
— Prepared either as _______ w/ [2] coupled with [+] _______ polymer=____________.

A

tincture

alcohol
iodophor

neutral
Povidone-iodine

108
Q

In Iodine, when used in compromised skin, like in _____ [high skin permeability] → possibility of _______application or [less/excessive] skin application could lead to
[5]

A

infants
mucosal
excessive

Neutropenia
Dermatitis
Goiter
Hypothyroidism
Liver function abnormalities

109
Q

IODINE
● Absorbing agent: [2]
● Can dffiuse thru _______.

A

charcoal
milk

breastmilk

110
Q

ANTISEPTICS:
____________
– Not used anymore

____________
– Not used anymore for ophthalmia neonatorum
– REPLACED by ____________

A

Mercury

1% silver nitrate
erythromycin