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1
Q

What is a yeast?

A

Single celled, reproduce by budding, produce moist, opaque, creamy colonies on media

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2
Q

What is a mold?

A

Multi-cellular, more complex than yeasts, looks fluffy, variable in colour

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3
Q

What is a Hyphae?

A

gives mold fuzzy appearance

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4
Q

What are the two types of Hyphae?

A

Vegetative and Aerial
Vegetative is submerged and on the surface
Aerial is above surface (fuzzy) and produces Conida (asexual spore)

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5
Q

4 Groups of identification of Fungi

A
  1. Aspergillus Niger - round ball, black appearance
  2. Penicillum - skeletal shape - most common - produces penicillin - found by Alexander Fleming
  3. Rhizopus - two sacs
  4. Tinea Corporis - Hyphae-like
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6
Q

4 Types of Infections caused by Fungi

A
  1. superficial - dead layer of skin
  2. Cutaneous - first layer, penetrate all keratinized tissue like hair, skin, and nails
  3. Subcutaneous
  4. Systematic - blood, bone, CSF, internal organ
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7
Q

Ideal temperature for Fungal Growth?

A

Ideal temp is RT, 23-25 degrees, not 37 degrees

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8
Q

Dermatophytes are?

A

skin causing infections

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9
Q

Cutaneous Infections are?

A

infect keratinized tissues of body, and cannot live in deeper tissue, caused by dermatophytes, refer to as “ringworms or tinea”

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10
Q

Tinea pedis is known as?

A

Athletes foot

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11
Q

Tinia corporis is?

A

Ring lesion

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12
Q

Tinea unguium is?

A

Thick, discoloured nail

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13
Q

Candidas Albicans is mainly in…?

A

Vaginal infections

Oral or vagina “thrush”, albicans = white

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14
Q

Mycosis is…?

A

infection caused by a fungi

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15
Q

Fungis are…?

A

Eukaryotes - true nucleus, nuclear membrane,

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16
Q

Specimen handing is composed of?

A

Collected aseptically, swab is not accepted except mouth, genital, external ear
tissue and body fluid preferred
collect nail/skin scarping in sterile black paper

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17
Q

Specimen collection is composed of?

A

Any tissue/body fluid can be cultured for fungi
Most species are respiratory secretion, others are hair, skin,nails, and scrapings.
Dermatophytes stored at RT, blood & CSF at 35 degrees, others can be refrigerated

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18
Q

Dimorphic means?

A

can grow in different conditions as fungi can be a yeast at 35 degrees and a mold at RT

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19
Q

Direct microscopy using KOH

A

most common for skin, hair, nails, and tissues.

10-15% KOH breaks down keratin, fungus not broken down due to chitin

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20
Q

Culture media for Mycological samples?

A

SAB - Saboured Dextrose Agar

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21
Q

Steps of Fluorescent Microscopy

A
  1. Energy absorbed by atom - excited
  2. Electron jumps to higher energy level
  3. Electron drops back to ground state, emitting a photons - atom is fluorescent
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22
Q

Principle of Fluorescent Microscopy

A

Material emits detectable visible light when irradiated with light of a specific wavelength.
Certain dyes called fluorochrome can be raised to a higher energy after absorbing UV light
When dye molecule returns to normal, energy is released as a form of light - fluorescence
Intensity of contrast is better than chromogenic dye of gram staining and light microscopy

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23
Q

Fluorochroming is…?

A

Uses fluorescent dye alone and a direct chemical interaction between dye and composition of cell, similar to light microscopy.

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24
Q

Iummunofluorescence is…?

A

Dye is linked (conjugated) to a specific antibody and can only stain specific cell with a specific antigen, specific antibody attached to dye and if the bacteria has the specific antigen, it will fluorescent.

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25
Acridine Orange is...?
non-specific, binds to nucleic acid, stain all nuclei, stain bright orange, confirm with blood culture, but is not selective between neg. and pos.
26
Auramine-Rhodamine
For mycobacterium culture - mycolic acids in cell walls have affinity for this and is a direct detection of mycobacterium, ONLY bacilli will stain, more specific than Acid Fast
27
Calcofluor White
enhance visibility of fungal elements, binds to chitin in cell walls, will cause fungi to fluorescent, easier to identify
28
KOH
destroys keratin, leaves fungal hyphae, can add chitin specific stain, stain lyses cells, doesn't stain, non-specific
29
Direct Immunofluorescence
direct immunofluorescence (IF) uses single aB directed against target of interest, primary aB directly conjugates to fluorophore
30
Indirect Immunofluorescence
uses two aB, primary aB is unconjugated, secondary fluorophore aB directed against primary aB used for detection
31
Four types of Fluorochroming
Acridine Orange Auramine-Rhodamine Calcofluor White KOH Prep
32
Types of Automation in Gram Staining in Micro
Previ Colour Gram AGS-100 Aeropray
33
What does Automatic ID/MIC do?
Continuous monitoring, results release when done, rapid TAT, some results in 3hrs, link to LIS so it is paperless
34
What does Vitek-MS do?
Uses mass spec - MALDI-TOF - Matrix Assisted Lazer Desorption Ionization - Time of Flight, it is easy, rapid, and reproducible.
35
Types of Automation in Micro?
Vitek Phoenix System Siemans Microscan Walkway
36
Procedure using Abscess-Deep
Anaerobic Transporter Within 24hrs/RT 24/RT BA, CA, MAC, ANA, THIO
37
Procedure using Blood or Bone Marrow
Culture Bottle Media Set (aerobic and anaerobic) Within 2hr/RT Incubate @37 degree
38
Procedure using Body Fluids
Sterile screw cap tubes or Anaerobic Transporters Immediately/RT Plate as soon as arrival BA, CA, MAC, ANA, THIO
39
Procedure using CSF
Sterile screw cap tubes Immediately/RT Plate as soon as arrival BA, CA, THIO
40
Procedure using Rectal Swab
Swab in enteric transport medium Within 24hr/4 degrees 72hrs/4 degrees MAC, SMAC, HEK/SS, Campy BA
41
Procedure using Ova & Parasites
O&P transporter - 10% formalin and PVA Within 24hr/RT Indefinite/RT
42
Procedure using Vaginal Swab
moistened with Stuart's or Amie's medium or JEMBEC transport medium Within 24hrs/RT 24hr/RT BA, CA, MAC, TM
43
Procedure using Sputum
Sterile screw cap container Within 24hr/RT 24hr/4 degrees BA, CA, MAC
44
Procedure using In-Dwelling Cetheter
Sterile screw cap container Within 2hrs/4 degrees 24hrs/degrees BA, MAC
45
What are Superficial Infections?
Mild infections, and are non-dermatophyte agents
46
What are Cutaneous Infections?
Infect keratinized tissues of body and cannot live in deeper tissue. Caused by dermatophytes which causes skin infections disease refer to ringworms or tinea
47
What are Systematic Infections?
blood, bone, internal organs, CSF, Urine and are associated with dimorphs
48
Bottom part of the Hyphae is referred to as?
Vegetative - submerged on surface
49
Top part of the Hyphae is referred to as?
Aerial - above - fuzzy - product of Conida
50
Round ball, gives black appearance for fungi
Aspergillus Niger
51
Skeletal shape for fungi
Penicillum
52
Two sac shape for fungi
Rhizopus
53
Hyphae-like for fungi
Tinea Corporis
54
CLED Medium
Cysteine Lactose Electrolyte Deficiency plate Differential, identify bacteria in urine Support growth or urinary pathogens & contamination Prevents swarming of Proteus species due to lack of electrolytes
55
EMB Medium
``` Eosin Methylene Blue Gram (-) isolation, similar to MAC NLF = clear LF = precipitate Lactose (+) = metallic green ```
56
Multiplier using 10uL loop
1:100
57
Multiplier using 1uL loop
1:1000
58
CLED Principle
Use lactose - fermenation Cysteine permits growth of dwarf colonies Bromothymol blue - pH indicator to differentiate Organism that ferments lactose will lower pH, change media from green to yellow. Electrolytes reduced to restrict swarms of Proteus species.
59
Coagulase test
S. aureus known to produce coagulase
60
Catalase test
Breakdown of H2O2 - most likely Strep species
61
Differentiating Growth on BA & MAC
``` If gram (+), catalaze and coagulase test done to differentiate S. aureus and streptococcus Gram stain to confirm ```
62
What is composed in Culture and Sensitivity Test?
Colony Count and Antibiotic Susceptibility Test
63
How many CFU colonies indicates an UTI infection?
>10^5 or 100,000 CFU/mL
64
How many CFU colonies indicate vaginal contamination?
<10^4 to 10,000 CFU/mL
65
How many CFU colonies needs to be re-evaluated?
Between 10^4 and 10^5 or 10,000 to 100,000 CFU/mL
66
Types of Specimen Transportation
``` Amie's or Stuart's transport media Cary-Blair medium Sterile containers Blood culture bottles Anaerobic transport media Syringes (no needles) for sterile biopsy fluid ```
67
Criteria for Transport Medium
Sterile Maintains viability of microorganism No Nutrients
68
Nose swabs can mainly be used for?
Staphylococcus aureus and whooping cough
69
Mouth swabs can be mainly be used for?
Yeast infections such as C. Albicans
70
Cary-Blair Medium is?
For rectal and fecal swabs - enhance viability of SS species, buffered to prevent pH shift, low in nutrients to inhibit species.
71
MAC
Isolate and differentiate Enterobacteriaceae Gram (-) selective Uses Lactose as energy Acid produced when lactose ferments, pH drops and causes a change in pH indicator + bile precipitates
72
Selective agents of MAC
Bile salts, oxgall, CV
73
Salmon-Shigella Plate
isolate SS species, contains lactose and Na thiolate as differentiating agent Neutral red as pH indicator
74
Selective agent of SS Plate
Bile salts and brilliant green dye
75
Salmon in SS plate gives what colour colonies and why?
Black colonies because Salmonella reduces HS producing H2S
76
Shigella in SS plate gives what colour colonies and why?
Non-black colonies - because it cannot reduce HS
77
What enables the detection of SS colonies?
Detection of HS from Na Thiosulphate and Ferric Citrate
78
Thayer-Martin Plate
Chocolate agar with antibiotics Isolates Neisseria bacteria such as gonorrhea and meningitis Consist of Vancomysin which kills gram (+), Collistin which kills gram (-) except Niesseria, but is not active against Proteus, and Nystatin which kills fungi
79
Alpha hemolysis
Zone of greenish clearing due to complete breakdown of hemoglobin such as Strep species or normal flora
80
Beta hemolsis
Clear zoning around colonies due to hemolysis such as Strep pyogens causing Strep Throat
81
Is Blood agar selective or differential?
Differential
82
Selective media allows...?
Certain types of organisms to grow, and inhibit the growth of other organisms.
83
Differential media allows...?
Qualitative changes on colony growth to target organisms, does not kill like selective, but indicate the presence of target organism

MLT (4 decks)

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