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BACTE STAINING-CONTROLS Flashcards

(110 cards)

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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
MODIFIED ACID-FAST STAINING METHOD
Modified Kinyoun
26
— ID of intestinal coccidian oocysts
Modified Kinyoun
27
— Ideal for cryptosporidia and cyclospora parasites
Modified Kinyoun
28
MODIFIED KINYOUN — Specimen : ______ — Rgt: = __________
stool conventional acid-fast
29
MODIFIED KINYOUN Result: ______ appear as [color]-stained organisms against a [color] background
oocysts magenta blue
30
If the result is: Bacteria: [Method] Parasite: [Method]
Acid-fast/Kinyoun Modified acid-fast/Modified kinyoun
31
Staining technique of the ff: Cell wall Capsule
Dyar Anthony's. Hiss and Gin's +Nigrosin
32
Staining technique of the ff: Metachromatic Granules Endospore
Neisser + Albert Domer+ Schaeffer-Fulton
33
Staining technique of the ff: Flagella Spirochetes
Gray + Leifson Levaditi's + Fontana-Tribondeau
34
Staining technique of the DNA:
Feulgen
35
process that [X] Defined scope of microorganisms including some spores
Disinfection
36
Disinfectant can be either?
Physical Chemical
37
[X] pathogenic bacteria so items are safe to handle or dispose of
Decontamination
38
WHY THE BACTERIA WON’T DIE?
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
39
[+]Spores
Calcium, Protein, Carbs, Lipids, Dipicolinic acid
40
Opposite for virus with lipid-rich envelope detergent + wetting agent.
Mycolic acid in bacteria
41
nonliving; abnormal infectious proteins
Prions
42
Prions resistant to: [3]
heat 121C radiation chemicals [acid/base]
43
- determines the exposure time that is necessary for 99.9% elimination
Microbial load
44
–- Preparation, dilution and use must be followed carefully by this [2 examples]
Disinfecting agent conc. ● 70% alcohol ● Povidone-iodine
45
NO disinfecting agent, full contact= presence of ________
organic material
46
__________________ Nature of the equipment may affect, so it is important to consider whether the material need to be ONLY: ________ or _________?
Nature of surface to be disinfected disinfected sterilized
47
CONTACT TIME –- Identify the ________ present first –- Contact time may ONLY ______ / _______ Betadine - ______ minutes
Mx disinfect/sterilize 1-2 mins
48
Disinfectant:_________°C. ● ____°F (____°C) = Killing activity
2O-22C 49F/ 65C
49
PH –- ______ of bacteria desired to be killed –- _______
ID Surface ID
50
BIOFILMS –- May need to [INC/DEC] CONC of _______ / _______ time
INC Disinfectant Contact time
51
COMPATIBILITY OF DISINFECTANT AND STERILANT — 2 disinfectants [ARE/ ARE NOT] better than 1 –- __________ may negate other disinfectant. ● Example
ARE NOT Disinfectants bleach & quarternary compounf
52
PHYSICAL Sterilization [5]
Incineration Moist Heat Dry Heat Filtration Ionizing Radiation
53
PHYSICAL DIsinfection [3]
Boiling Pasteuring Non-Ionizing radiation
54
CHEMICAL Sterilization [2]
Ethylene Oxide Hydrogen Peroxide
55
CHEMICAL Sterilization/Disinfection [4]
Alcohol Antiseptics Acetic Acid Phenolics Hypochlorite Quarternary Ammonium Compound
56
INCINERATION –- Burned to ____ [____-____] — Good for ______ — ______ method CONS: [2]
87O-98OC prions SAFEST toxic air emissions [+] heavy metals
57
DRY HEAT — [Shorter/Longer] exposure time — Done for ____ hours — [TEMP] Used for [cold/heat-stable] substance such as [2]
Longer 1.5-3hrs 16O-18OC heat: porcelain/glassware
58
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
1 atm/ 15 pounds 121C 4 Faster surface Bacillus stearothermophilus Biological indicator
59
MOIST HEAT ●Culture Media: - ____ °C= 30 mins - ____ °C= 4 mins ● Biological waste: - 132 °C= ______
121 132 30 - 60 mins
60
Moist heat [can also/ cannot] be used for prions
Can also
61
Steps in Moist Heat [6]
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
Liquid Filtration - uses [3]
cellulose acetate nitrate membrane vacuum
63
Filtration Of Bacteria,Yeast,Molds - Uses _____um pores of membrane filter - Range: - [X] # bacteria/fungi but NOT _____
0.45um 0.2-0.45um virus
64
Uses 0.22um membrane filters for parenteral solutions + alcohol
critical sterilizing
65
[X] vegetation cells + spores but NOT virus
critical sterilizing
66
uses HEPA filter - [X] >0.3um sized-Mx
Air filtration
67
under moist heat, it is the fastest and simplest, all organisms + spores [except for prions]
autoclaving
68
Autoclaving is operated by a laboratory technician. Before opening, make sure that the pressure or PSI is completely_____.
0
69
principle is steam under pressure
Autoclaving
70
moist heat/autoclaving biological indicator
Bacillus stearothermophilus
71
Autoclaving for media, liquids, pipettes, utensils, and etc. [Temp] [psi] [time]
121C 15 psi= 15-3Omins 132C= 4mins
72
Autoclaving for biological wastes [Temp] [psi] [time]
132C 15psi= 3O-6Omins
73
Indication of the ff in Bacillus stearothermophilus: Black Masking tape Black Stripes
exposed to extreme convention sterilized
74
— made up of fibrous or granular materials Example [3]
Depth Filters Berkefield/Chamberland filter Abestos
75
__________filter for Porous membranes [0.1 um thick] [1 example]
membrane filters filter paper
76
cellulose acetate, polycarbonate
membrane filters
77
* sterilize pharmaceuticals, ophthalmic solution, culture media, antibiotics and oil products
membrane filters
78
____________ * bacteriostatic * [-] metabolism rate * Important in food microbiology * Blood products
Low/Cold Temperature
79
__C -__C for ___ hours – kills syphilis
2 8 72
80
disruption of metabolism involves removing of water from microbes.
Dessication
81
protein changes + DEC. chemical reaction. * Powdered form
Lyophilization
82
RADIATION-IONIZING –- [Hot/Cold] Sterilization — Causes ____ mutation +_______ prod. — destroys [2] Biological Indicator: - __________
Cold DNA Peroxidase vegetative endospores of both prokaryotes & eukaryotes Bacillus pumilus
83
Radiation Ionizing --- Gamma rays [1500 to_______ radiation] & Xrays
2500
84
Best for: - gloves, plastic syringe, catheters
Radiation-Ionizing
85
BOILING — ____ °C for ___ mins. — kills only _______ bacteria [sporulating/non-sporulating]. * [Example]
1OOC 15mins vegetative non-sporulating Water bath
86
PASTEURIZATION — ____°C for ____ mins. — kills ______ pathogens +Mx W/O DEC. ______ nutrition or _____ for _______. — Sterilize [3] — CANNOT eliminate bacterial _________ * [Partial/Total] Sterilization
7OC 3Omins food food flavor spoilage milk, dairy products, OH endospores
87
TYNDALIZATION —_____°C for ____ minutes for ___ consecutive days — kills [2] — __________ Sterilization. * Example: _______ sterilizer [free-flowing steam]
1OOC 3O mins vegetative, endospores Fractional/Intermittent Arnold's
88
NON-IONIZING RADIATION — LONG ________ LOW ______ — [penetrate/do not penetrate] well —- Damage to cellular ____ by producing _______.
wavelength energy do not penetrate DNA thymine dimers
89
— Used on: ---- exposed surface + operating rooms ---- gloves, plastic syringe, catheters
Non-Ionizing Radiation Ionizing Radiation
90
— # chemical sterilant
Ethylene oxide
91
Used for sterilizing heat-sensitive objects
Ethylene Oxide
92
CONS: lengthy cycle time + potential health hazardq
Ethylene Oxide
93
–-used if stabilized w/ phytate, citrate or malonate.
Hydrogen peroxide
94
This form of Hydrogen peroxide is used to sterilize HEPA filters
Vapor form
95
It has NO toxic by-products
Hydrogen peroxide
96
–- Non-sporicidal and evaporates quickly.
Alcohols
97
— As antiseptic/ thermometer /injection vial rubber septa :limited to the skin
Alcohols
98
Alcohols' Water content– hydrolyzes ______ in bacterial protein
H bonds
99
— ALL Mx-cidal + sporicidal
Hypochlorite
100
HYPOCHLORITE — _____ ratio [bleach to water] — Inexpensive yet ______. — [X] DEC. capacity based on quality of ______. CONS: –[2] [3 physiologic] + –addition with [2] = toxic gas
1:1O effective water corrodes metal discolors fabrics oropharyngeal irritation esophageal irritation ocular irritation ammonia/acids
101
ALDEHYDE — [X] [3] –- __________Technique Example: _________
corrode lenses rubber metals Cold Sterilization Technique Glutaraldehyde
102
2 CHEMICAL METHODS STERILIZATION / DISINFECTION that use Cold Sterilization Technique
Aldehyde Periacetic Acid
103
it is best used if the equipment has organic material
Periacetic acid
104
HEAVY METALS — [X] microorganisms by ________________. -– [5 metals examples] — __________: eye drop for Neisseria gonorrhoeae. — __________: antiseptic.
inactivating/precipitating cell proteins Copper Arsenic Mercury Silver Zinc AgNO3/Silver Nitrite Mercuric chloride
105
— 1ST widely used antiseptic/ disinfectant
Phenols
106
PHENOLS -- ______-% Phenol - ____ to ____ mins contact time –- Effective against ________ — [X] [2]
5 1O-3O mins Mycobacteria plasma membrane protein
107
IODINE — Prepared either as _______ w/ [2] coupled with [+] _______ polymer=____________.
tincture alcohol iodophor neutral Povidone-iodine
108
In Iodine, when used in compromised skin, like in _____ [high skin permeability] → possibility of _______application or [less/excessive] skin application could lead to [5]
infants mucosal excessive Neutropenia Dermatitis Goiter Hypothyroidism Liver function abnormalities
109
IODINE ● Absorbing agent: [2] ● Can dffiuse thru _______.
charcoal milk breastmilk
110
ANTISEPTICS: ____________ -- Not used anymore ____________ – Not used anymore for ophthalmia neonatorum – REPLACED by ____________
Mercury 1% silver nitrate erythromycin

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