Cytology Lab

Front 1

Reporting Yeast

Back 1

Document how many (budding vs. non), bacteria observed (type and amount), and cells (blood vs. epithelial) form the entire slide;

Don't forget which ear

Front 2

Reporting on Otododectes

Back 2

Count the number of mites and eggs you see per slide;

Don't forget which ear

Front 3

Lyme Disease

Back 3

Caused by the bacteria: Borrelia burgdorferi;

Spread by hard ticks (Ixodes scapularis);

When removing a tick, ensure you remove the head because it may cause an abscess

Front 4

Volume of Ejaculate

Back 4

Measured with a volumetric flask;

Total ejaculate for a bull is usually 5mls (male cats= 0.04mls);

Ejaculate volume is larger than spermatozoa and varies amongst collection methods;

Three components: sperm free fraction, sperm rich fraction, and sperm poor fraction

Front 5

Gross Appearance

Back 5

Opacity (reflects spermatozoa concentration);

Words to describe: Thick, creamy, opaque, milky opaque, opalescent milky, watery and white (normal bulls- opaque/creamy/white) and discoloration due to contamination

Front 6

Sperm Motility

Back 6

If correlated with fertility!;

Improper specimen handling adversely affects the motility;

2 assessments: wave motion, and motility

Front 7

Wave Motion

Back 7

Gross motility based on swirling activity observed in a drop of semen on a microscope slide at low magnification (4X);

Very good= vigorous swirling;

Good= moderate slow swirling;

Fair= barely swirling;

Poor= no swirling;

Bulls have high sperm density so should show vigorous swirling;

Wave motion is decreased if sperm concentration is decreased

Front 8

Motility

Back 8

Diluted drop of semen with warm physiological saline with a coverslip and observed at 10X;

1 drop of semen on slide, dilute with saline to produce a monolayer, place coverslip;

Subjective:

Very good= rapid linear activity (80-100%);

Good= moderate linear activity (60-80%);

Fair= slow linear or erratic activity (40-60%);

Poor= very slow, erratic activity (20-40%);

A satisfactory sample should have at least 60%

Front 9

Sperm Concentration

Back 9

Diluent is formalin, Sodium bicarb and distilled water;

Take 0.1mls semen and add 9.9mls diluent;

Gently mix sample until homogeneous;

Place semen on a Hemacytometer and allow to sit for 2-3 minutes;

Count both grid areas at the 10X (withing 10%?- take the average);

Multiply the number b 1000000 and this will result in a # sperm per ml of the diluted semen sample;

Front 10

Normal Sperm Concentration for a Bull

Back 10

1200 million/mL (range 300-2500 million/mL)

Front 11

Live:Dead Sperm Ratio Procedure

Back 11

Have microscope slides and stain prewarmed to body temperature;

Pipette a drop of stain (negrosin/eosin mixture) onto the end of a slide, then pipette a small droplet of semen next to the stain;

Rock that slide back and forth a few times to mix the sperm and stain;

Smear like a blood smear;

Dry the slide rapidly;

Examine using 100X objective lens

Front 12

Live:Dead Sperm Ration Count

Back 12

Observe at 100X and count 200 cells;

result expressed as a percent of live sperm;

Live sperm resist staining, dead sperm pick up the eosin and appear pink, the background should appear blue

Front 13

Sperm Morphology

Back 13

Observe 100-500 cells;

Abnormalities are divided into:

Head, midpiece, and tail;

Generally 20% of sperm are abnormal

Front 14

Sperm Morphology: Primary

Back 14

Serious abnormalities;

Occur during production;

Head too big/small, doubled, oddly shaped;

Midpiece swollen, kinked, double, twisted, essectrically attached to head;

Tails that are coiled

Front 15

Sperm Morphology: Secondary

Back 15

Storage in the epididymus to the smear;

Tailless heads;

Bent or broken tails;

Protoplasmic droplets on midpiece

Front 16

Back 16

Misshapen Head

Front 17

Back 17

Elongated Head

Front 18

Back 18

Double head

Front 19

Back 19

Double Tail

Front 20

Other cells in sperm...

Back 20

Normal semen has few: WBCs, RBCs, and epithelial cells;

NEVER has: bacteria and yeast

Front 21

Formal Evaluation

Back 21

Signalment;

Volume;

Gross appearance;

Wave motion;

Motility;

Concentration;

Live:Dead Sperm Ratio;

Morphology

Front 22

Normal Total Sperm Ejaculate for a Bull

Back 22

4-5 billion

Front 23

Normal Morphology for a Bull

Back 23

Greater than 70% normal

Front 24

Back 24

Anestrus:

Non-cornified squammous epithelial cells, may contain some neutrophils, no RBCs

Front 25

Back 25

Proestrus:

Cornified squammous epithelial cells replace non cornified, neutrophils decrease, and RBC's increase

Front 26

Back 26

Estrus:

Cornified squammous epithelial cells, no neutrophils, RBCs present in small numbers, neutrophils increase as diestrus approaches

Front 27

Back 27

Diestrus:

Cornified epithelial cells are replaced by non cornified, abundant cytological debris, neutrophils increase in number by the 3rd day of diestrus and decrease to few by about the 10th day, no RBCs

Front 28

Cytology Report: Lower Power (10X)

Back 28

Scan for large objects, cell clusters, parasites, fungal hyphae and crystals

Front 29

Cytology Report: High power (40X)

Back 29

Determine prominent cell type

Front 30

Cytology Report: Oil immersion (100X)

Back 30

Describe cellular characteristics

Front 31

Imprints

Back 31

For Muscle Tissue Samples:

Done on external lesions or from removed tissues;

Easy to collect;

Collects fewer cells;

Greater contamination;

Disadvantages: few cells collected, cells obtained from the surface of a lesion may not be representative of the whole lesion;

Front 32

Procedure for making Imprints

Back 32

Imprint dirty then clean; Make several dirty imprints;

Blot tissue on clean, wet (saline), absorbed material to remove fluid;

Dry lesion;

Make several clean imprints (do not drag);

Dry the slide quickly;

Stain with dirty diff quick;

Observe under 40X and oil;

Front 33

FNA- Adipose Tissue (small mass)

Back 33

21g needle and a 12cc syringe;

Alcohol skin prep;

Mass held securely;

Needle introduced in center of mass;

Strong negative pressure applied (3/4 of syringe);

Sample several areas (avoid surrounding tissue);

Relieve negative pressure before removing;

Several squash preps are made;

Dry quickly and stain with dirty diff quick;

Observe under 40X and oil

Front 34

FNA- Adipose Tissue (large mass)

Back 34

21g needle and a 12cc syringe;

Alcohol skin prep;

Needle stays within mass;

Redirect to other areas;

Negative pressure relieved during redirection and removal;

Several squash preps are made;

Dry quickly and stain with dirty diff quick;

Observe under 40X and oil

Front 35

Non-Aspirate- Bone Marrow

Back 35

Mass held firmly;

Needle with syringe attached is introduced in mass (syringe pre-filled with air);

Needle is moved rapidly back and forth through the mass five to six times along the same tract;

Cells are collected by shearing and capillary action;

Needle removed from mass, material is expelled by depressing plunger;

Repeat in more than one site and create several slide;

Squash prep and air dry quickly;

Stain with dirty diff quick;

Observe under 40X and oil

Front 36

Hair Pluck

Back 36

Grasp hair close to the base and remove in the direction of the follicle;

Place on microscope slide and confirm you did not break the hair and the follicle is intact;

Positive for ringworm- fungal spores attached to the hair shaft (40% accurate)

Front 37

Culture Swabs

Back 37

Use to isolate bacteria and viruses;

Areas that can be swabbed include:

Mucus membranes of the urethra, cervix, rectum, eye, throat, vagina, and penis;

Fluid collected from thoracic or abdominal cavity;

And non purulent wounds;

It is important to follow asepsis in order not to contaminate the sample and to ensure you have the appropriate aerobic/anaerobic medium for the bacteria being tested

Front 38

Procedure for Culture

Back 38

Ensure appropriate sampling (aerobic vs. anearobic or both);

Aseptically handle all materials;

Peel back plastic wrap exposing the caps;

Open tube and discard plug;

Take sample with swab;

Insert swab into tube and secure cap

Front 39

Post Sampling for Culture

Back 39

Refrigerate labeled (Animal ID, site, date and time cultured on each sample) transport media vials containing swabs (do not freeze);

Send with refrigerated transport packs in a styrofoam shipper so that culture is set-up within 48hrs of taking the swab- older samples will be rejected as they are unfit;

Cultures generally take 7 days fom the time of placing the material on the selective culture media;

Negative status cannot be confirmed before 7 days;

Identification of fungal or bacterial contamination which would preclude reading of the test, will be done as early as possible so that re-sampling and resubmission can occur

Front 40

Canine Heartworm Antigen Test Kit

Back 40

Snap test used to test for the presence of adult D. Immitis antigens in dog or cat blood;

Front 41

ERD Health Screen Feline Urine Test

Back 41

Detects level of albumin in feline urine;

A test used to determine if a cat has damaged nephrons due to a variety of reasons

Front 42

Canine Pregnancy Test Kit

Back 42

Determines pregnancy by measuring relaxin levels in plasma or serum;

Valid 20-31 days post LH surge;

Front 43

Back 43

Hymenolepis spp.;

Tapeworm

Front 44

Back 44

Syphacia spp.

Pin worm;

Inhabit the cecum and colon of rats and mice;

Nonpathogenic in immunocompeten rodents;

In labs, infections may have effects on behaviour, growth, intestinal physiology, and immunology;

Effective pinworm surveillance and eradication vital in research animal facilities;

Diagnosed by the use of perianal tape impression

Front 45

Back 45

Aspiculuris spp.

Pin worm;

Found in mice and rats;

Located in the colon and cecum

Front 46

Back 46

Cryptosporidium spp.

Protozoa;

Causes diarrhea;

Fatal in immunocompromised;

Thick fecal smear

Front 47

Back 47

Eimeria spp.

Live in the small intestines;

Diagnosed by fecal float;

Can be sporulated or unsporulated

Front 48

Back 48

Trichosomoides spp.

Larvae develop in the striated muscle;

Migrate to he nasal mucosa;

Worms can be found between the incisors in the mucosa;

Under natural conditions, the inflammation of the nasal mucosa that is induced by the parasites might reduce the competitiveness of infected rodents when foraging or looking for potential mates

Front 49

Back 49

Spironucleus spp.

Small symmetrical, flagellated protozoa;

Found in the intestinal tract;

Most common in fish

Front 50

Back 50

Dentostomella spp.

Pin worm of gerbils;

Causes diarrhea;

Fecal float

Front 51

Back 51

Notoedres spp.

Scabby, scaly facial disease known as head mange;

Highly contagious;

Similar to sarcoptes but smaller and rounder

Front 52

Back 52

Tixacarus cavia;

Guinea pigs;

Affects the head, shoulders, back, and flanks of the guinea pig, may spread;

Secondary bacterial infections can occur

Front 53

Back 53

Psoroptes spp.

Rodent and rabbits;

Scraping or exudate from ears

Front 54

Back 54

Cheyletiella spp.

Rabbits;

"Walking dandruff"

Direct observation, mites usually direction surface of skin;

Excessive dandruff

Front 55

What sample do you require for a Canine Pregnancy Test?

Back 55

Serum or Plasma collected in an anti-coagulated with EDTA or Sodium Citrate

Front 56

How many times should the Canine Pregnancy Test be performed and in what time interval, to confirm a negative result?

Back 56

The test should be performed twice, one week apart in order to confirm the status

Front 57

Describe a negative and positive result of the Canine Pregnancy Test

Back 57

Positive= one pink/purple band in both window 2 & 3;

Negative= No band reading in the #2 window but there will be a band in #3

Front 58

What does the Heartworm Antigen Test Diagnose?

Back 58

Dirofilaria immitis antigen

Front 59

Why would we perform the Heartworm test?

Back 59

Routine testing

Front 60

What would a negative test result look like for Heartworm?

Back 60

Negative= only the positive control spot develops

Front 61

What would a positive test result look like for Heartworm?

Back 61

Any color development in the sample spots as well as the positive control

Front 62

When would you deem the test invalid for Heartworm?

Back 62

If there is background color, and if the positive control spot does not appear

Front 63

What does the ERD Health Screen diagnose?

Back 63

Detection of microalbuminuria in cats

Front 64

When would you test a patient with the ERD Health Screen?

Back 64

Health screen and evaluation for early renal damage is indicated

Front 65

What test do you perform before proceeding with the ERD? Why?

Back 65

Prior to performing the ERD, you must obtain the urine specific gravity. The result will determine where to place/add urine in the test and whether to dilute the sample.

Front 66

Name 4 diseases that damage nephrons allowing albumin to leak into the urine

Back 66

Inflammatory disease;

Infectious disease;

Metabolic disease;

Neoplasia

Front 67

What type of sample do you require for a Canine Parvo Antigen Test and how would you collect it?

Back 67

You need a fecal sample collected directly from the anus

Front 68

What form of CPV is prevalent in puppies? Why not the other form?

Back 68

Enteritis is prevalent in puppies because the myocardial form is rare

Front 69

Once the pouch is open, the CPV test should be used within how many minutes?

Back 69

The test should be completed within 10 minutes

Front 70

What does a positive result look like for CPV?

Back 70

Once pink/purple band in window #2 and window #3;

Front 71

What would an invalid result for CPV look like?

Back 71

There will be no pink/purple band in window #3

Front 72

Transudates

Back 72

Fluid that passes through a membrane which filters out much of the protein and cellular elements to yield a watery solution;

Due to increased pressure in the veins and capillaries pressure forcing fluid through the vessel walls or low levels of protein the blood serum - a filtrate of blood;

causes edema;

SG: <1.017;

TP: <3g/dL

Front 73

Transudates commonly found in...

Back 73

Acites or pleural effusions found with heart failure or other circulatory disorders

Front 74

Modified Transudates

Back 74

Transudate with additional protein and/or cells;

May be in a transitional stage progressing to an exudate;

Moderate cellularity;

TP: 2.5-7.5 g/dl;

SG: 1.017 or less than 1.025;

Sediment shows mesothelial cells and macrophages;

Amber or pink and turbid

Front 75

Exudates

Back 75

Fluid rich in protein and cellular elements that oozes out of blood vessels due to inflammation and is deposited nearby;

Altered permeability of blood vessels permits the passage of large molecules and solid matter through their walls;

Increased cellularity;

TP: >3g/dl;

SG: >1.025;

Sediment shows neutrophils, lymphocytes, and small number of macrophages;

Higher cell count and protein value are indicative of inflammation or injury;

ex: Perspiration, pus, and serum;

Further classified into : septic or non septic and degenerative or non-degenerative

Front 76

Back 76

Septic exudate;

Neutrophils with intracellular bacteria

Front 77

Non septic Exudate

Back 77

Sediment does no show intracellular bacteria

Front 78

Degenerative Exudate

Back 78

Sediment shows nuclear changes such as pyknosis, karyorrhexis, and karyolysis

Front 79

Non-degenerative Exudate

Back 79

sediment does not show nuclear changes

Front 80

Modified Centrifuged Buffy Coat Smear

Back 80

Used to concentrate WBCs and platelets when looking for microfilaria, amastogotes (leishmania), erhlichia, and other blood parasites and mast cells

Front 81

Blood Typing

Back 81

Determine the patient's blood type before a transfusion;

Canine tests for DEA and cats for A & B blood groups

Front 82

Canine Blood type

Back 82

DEA 1.1 positive= RBCs carry that antigen;

Donors are usually DEA 1.1 negative or match the recipient;

If DEA 1.1 negative recipient is given DEA 1.1 positive blood, the recipient may develop antibodies and destroy the RBCs during the second transfusion

Front 83

Feline Blood type

Back 83

No universal donor among cats as they have natural antibodies against the blood antigen that they lack;

Cats are typed to match the donor and recipient;

Even in the first incompatible transfusion with cats, there is a transfusion reaction;

90% of cats are type A

Front 84

Crossmatching

Back 84

Always done prior to transfusion to assess donor and patient serologic compatibility;

If not compatible and blood is administered, the patient will have a hemolytic transfusion reaction (rapid destruction of donor red cells);

Crossmatch does not check for incompatible WBC or platelet antigen so reactions are still a risk with any transfusion

Front 85

Major Crossmatch

Back 85

This compares donor erythrocytes to recipient plasma to test for antibodies;

You are checking for preformed (aquired or naturally occurring) antibodies in recipient plasma against donor erythrocytes

Front 86

Minor Crossmatch

Back 86

This compares donor plasma to recipient erythrocytes and checks for preformed antibodies in donor plasma that could hemolyze recipient red cells;

Usually the donor plasma is markedly diluted after transfusion and is unlikely to produce a significant transfusion reaction, most often done in patients who have already had a transfusion

Front 87

Occult Blood Test on Fecal Sample

Back 87

Reagent tablets are used for detection of RBCs and/or hemoglobin in feces;

The procedure relies on the peroxidase-like activity of hemoglobin to enhance the oxidation of a chromagen in the tablet to orthotoluidine, which has a blue colour;

Small quantity of fresh feces is smeared onto a filter paper, regeat tablet is placed on top, wait 5-10 seconds, if blue = positive;

The test reagent reacts with myoglobin of muscle, carnivores should be fed a diet free of raw meat at least 3 days before testing so this doesn't result in a false positive

Front 88

Fecal wet mounts identify...

Back 88

protozoas (Giardia + Spirochetes), and purposful movement

Front 89

Fecal Smears identify...

Back 89

Clostridium;

Spirochetes;

Campylobacter- confirm with a culture;

Cryptosporidium - diagnose with acid-fast stain