Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

FISH 241 > Chapter 4 - Diagnostics > Flashcards

Chapter 4 - Diagnostics Flashcards

1
Q

what is the purpose of ‘Observation’?

A

recognize abnormal behaviour, physiology as indicators of possible pathogen or disease

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

what would you look for in your observations?

A 
behavior
-spinning, flashing
color
external lesions
internal pathology
bleeding
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

can you use observation as sole method for diagnosis?

A

nope

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What is the purpose of Microscopy?

A

-magnify image of specimen or host tissues for examination and identification

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

what are the two types of electron microscopy?

A

SEM: scanning
TEM: transmission

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

how do you manipulate the following for microscopy observation?

  • Viruses
  • Bacteria
  • Fungi
  • Protists
  • Histology
A 
V: need EM
B: Gram stain
F: Wet mount + Stain
P: wet mount + Stain
H: stain fish tissues to detect cell and tissue damage.
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

what is the purpose of Culture Techniques?

A

demonstrate pathogen is present and viable by growing it.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

four media types for culture techniques

A
  • general
  • selective
  • enrichment
  • diferencial
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

for Culture Techniques, what is API 20E?

A

20 biochemical tests condensed on one strip (differential media)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

what sort of media would you use for viruses?

A

Live media (cell culture): use living cells as growth medium.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

CPE. name, what does it do

A

Cytopathic effect.
Concept: pathogen destroys cells if present.

Use: Viruses

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

give some example types of live media

A
  • CHSE: chinook salmon embryo
  • SHK: salmon head kidney
  • RTG: rainbow trout gonad
  • FHM: fathead minnow
  • SSN-1: striped snakehead
  • EPC: epithelioma papillosum cyprini
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

CHSE:

A

Chinook salmon embryo. LIVE MEDIA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

SHK:

A

Salmon head kidney. LIVE MEDIA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

RTG:

A

Rainbow trout gonad. LIVE MEDIA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

FHM:

A

fathead minnow: LIVE MEDIA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

SSN-1:

A

striped snakehead. LIVE MEDIA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

EPC:

A

epithelioma papillosum cyprini: LIVE MEDIA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

What is the purpose of Serological Techniques?

A

detect presence or part of pathogen by using commercial antibodies to attach to pathogen

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

what do serological tests do?

A

detect antigens in sample or detect specific antibodies in the fish

21
Q

FAT: acronym

A

fluorescent antibody technique. SEROLOGICAL TECHNIQUE

22
Q

IFAT: acronym

A

Indirect Florescent Antibody Technique.

SEROLOGICAL TECHNIQUE

23
Q

ELISA: acronym

A

Enzyme Linked Immunosorbent Assay

SEROLOGICAL TECHNIQUE

24
Q

describe FAT (5 points)

A

fluorescent antibody technique

1: commerical antibodies (Ab) prepared by injecting lab rabbit with fish pathogen
2: rabbit produces Ab to fish pathogen, then a fluorescent dye is added.
3: Obtain sample of fish tissue suspect with pathogen, smear on slide
4: Add commercial labeled rabbit Ab to slide, then rinse
5: If pathogen present, stick to Ab and reacts with Ab. see glow, using flourescent microscope.

25
Q

describe IFAT (7 points)

A

indirect fluorescent antibody technique

1: rabbit blood injected into goat, goat produces Ab against rabbit blood, a florescent dye is added
2: rabbit injected w fish pathogen, rabbit produces Ab to fish pathogen (commercially prepared)
3: Obtain sample of fish tissue suspect w pathogen, smear on slide
4: Add rabbit Ab to slide, then rinse
5: Add labelled goat Ab to slide, then rinse
6: If pathogen present, sticks to rabbit Ab and reacts with rabbit Ab, then goat Ab attach to rabbit Ab- see glow
7: With indirect method, get enhanced signal and sometimes more cost effective.

26
Q

describe ELISA (5 points)

A

enzyme linked immnosorbent assay

1) Plate coated with a capture antibody, rinse
2) sample added, any antigen present binds to capture antibody, rinse
3) Detecting antibody added and binds to antigen, rinse
4) Enzyme- linked secondary antibody added, binds to detecting antibody, rinse
5) Substrate added, converted by enzyme to detectable form = glow or colour

27
Q

3 types of Commercial Antibody Kits. Why use these? How do you tell if they’re working?

A
  • Serum Agglutination
  • Latex Agglutination
  • Co-agglutination Test (antibody-sensitized staphylococci)
  • > real simple kits. good for a ‘quick test’ then you would do more samples or send off to lab.
  • > if the target organism is there, you get clumping. If not, smooth. Can be variable
28
Q

What is the purpose of Genetic Techniques?

A

detect a specific genetic code of the pathogen or its antigenic component.

29
Q

What does Genetic Techniques DO?

A

provides genetic signal, but not necessarily mean pathogen was viable and virulent.

30
Q

what sort of technique is PCR?

A

genetic technique

31
Q

PCR: acronym

A

Polymerase Chain Reaction

32
Q

what does PCR do?

A

amplifies target genome (via thermal cycling) to produce a copy large enough to be detected. 20 cycles= 10^6 copies of target DNA

33
Q

what are the three steps of PCR?

A

1) DENATURING @ 94-96C 20-30s DNA is split via melting
2) ANNEALING @ ~ 65C (20-40s) DNA target binds to original DNA strand
3) ELONGATION at 72*C (~ 1000 bp min^-1) DNA target is copied.

((heating, cooling, replication))

34
Q

name two types of PCR

A

RT-PCR: reverse transcription polymerase chain reaction (for RNA viruses)
Q-PCR: quantitative, real-time PCR measures amount of DNA amplified over times

35
Q

what does Sensitivity mean? (interpreting diagnosis tests)

A

% true positive.

reflects how well tests perform among group of DISEASED fish

36
Q

what does Specificity mean? (interpreting diagnosis tests)

A

% true negatives

reflects how well tests performs among a group of NON-DISEASED fish

37
Q

what are both sensitivity and specificity dependant on?

A

prevalence and number of fish sampled.

38
Q

what happens to specificity in moribund (almost dead, close to dead) fish?

A

secondary invaders can mask primary pathogen. specificity DECREASES

39
Q

what are the implications for ELISA when measuring specificity vs sensitivity?

A

It measures ANTIBODY response.
false positive: prior exposure.
false negative: no measurable antibody response

40
Q

what are the implications for PCR or RT-PCR when measuring specificity vs sensitivity?

A

It measures GENETIC code

  • false positive: genome present, but inactive
  • false negative: genome present, but insufficient quantity
41
Q

What is the implication for Culture Media when measuring specificity vs sensitivity?

A

Growth

-false negative. no growth, expired reagents, temperature, fastidious species, contamination.

42
Q

how would you test for covert (hidden) infections?

A

you could stress your fish and see if disease occurs. separate the stock, crank up the heat, wait and see.

43
Q

what do all diagnoses start with?

A

Observation

44
Q

when beginning your diagnosis, how should you get to know your fish?

A

check records

45
Q

what is adequate for diagnoses of many parasites, but not sufficient for viruses and most bacteria?

A

microscopy

46
Q

what method of diagnoses is very important for viruses and bacteria?

A

Culture

47
Q

how is culture important for viruses and bacteria?

A

demonstrates viability

48
Q

what sort of techniques are great for commercially important pathogens?

A

Serological techniques

Genetic techniques

FISH 241 (18 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions