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Front 1

Sample Preparation

Back 1

Collection after period of rest as it is more likely to be concentrated;

Refrigerate if the sample cannot be analyzed within 20-30 minutes (room temperature results in bacterial growth, chemical breakdown and cell lysis);

Thoroughly mix the sample, transfer the 5 mls to a conical centrifuge tube and centrifuge at specific requirements;

Once centrifuged, discard the supernatant;

Resuspend the sediment in remaining supernatant;

Using a pipette, transfer a small drop of urine to a clean microscope slide and apply a cover slip

Front 2

Stained Sample

Back 2

Sedistain;

With the remaining sediment left over after your unstained sample has been placed on a slide and add one drop of sedistain;

This can be read immediately or can sit for no more than 10 minutes;

Collect one drop of sediment with a pipette and place on the slide, and add the cover slip and analyze

Front 3

Microscopic Examination of Urine Sediment

Back 3

Usually done with low light, partially closed iris and with the condenser down;

Phase-contrast microscopes are a little more ideal with urine- compound microscopes with 2 added elements: a diffraction plate and a specialized condenser diaphragm, this allows details of objects by differences in refractive index

Front 4

Microscopic Examination of Sedistain Urine

Back 4

Stain can create artifacts, add bacteria and can precipitate in time which can be mistaken for bacteria;

Useful in differentiation;

Front 5

Method of Examination

Back 5

First scan the complete slide on 10X objective for large elements: casts, crystals and report in # per LPF;

Scan the entrie coverslip because some elements are pushed to the sides;

The remainder of the exam is done on 40X objective and finding are reported in # per HPF;

Bacteria and sperm described as mild, moderate and marked;

Report the morphology of bacteria;

Collection method is always reported because it affects the cells seen and is considered in final diagnosis

Front 6

Stain Precipitation

Back 6

Can give misleading impressions of bacterial presence

Front 7

Cytology Preparation of Urine Sediment

Back 7

Another approach to cell differentiation;

Used when abnormal or suspicious finding are seen on unstained sediment;

Helps to further differentiate or quantify bacteria;

In neoplastic situation, assess cells for malignancy

Front 8

Procedure of Cytology Prep of Urine

Back 8

Prepared by putting a small drop of sediment on the slide near the frosted end and the sample is spread with another slide usually in a squash, technique, air dry and stain with Dif Quick;

Some elements will wash off staining due to low protein nature of the specimen so serum-coated slides can be used to improve the sticking to the slide

Front 9

RBCs in Urine Sediment

Back 9

RBCs are smaller than WBCs;

Up to 3 to 5 per HPF- normal;

Usually pale yellow in colour;

Can be colourless if hemoglobin is diffused;

Are round, slightly refractile, and no internal structures, uniform and in shape and size;

Watch not to confuse with fat globules and yeast;

Can be confused with fat droplets;

Fat droplets will float in and out of focus, RBCs do not;

Fat droplets vary in size, do not take on a crenated appearance and will stain shades of orange in Sudan III of IV stain;

If still unsure: can add a small amount of 2% acetic acid to the slide and if the structures disapear, then they were RBCs;

Excessive numbers is referred to as Hematuria;

Hematuria can indicate bleeding somewhere in urogenital tract or trauma from expression of cystocentesis;

Can be due to trauma, calculi, infection, and neoplasia

Front 10

RBC Highly Concentrated Samples

Back 10

RBC's shrink and crenate;

Crenated- look darker, may look granular because of membrane irregularity

Front 11

RBC Dilute or Alkaline pH Urine

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RBC swell and may lyse;

Smooth edges look bigger;

May appear as colourless rings (ghost cells) that vary in size;

Can dissolve and not seen at all

Front 12

WBCs in Urine Sediment

Back 12

Round and granular, larger than RBCs but smaller than epithelial cells;

Most are neutrophils;

Normal very few are found;

Increases show active inflammatory condition or can be a contaminant;

Excessive called pyuria or leukocyturia and indicates infection of kidneys or bladder;

Always not presence of bacterial;

Seen with degenerated old urine;

May lyse in low concentrated or alkaline urine

Front 13

Squamous Epithelial Cells In Urine

Back 13

Derived from distal genital area (urethra, vagina, and vulva);

Finding bacteria and squamous cells together indicate contamination from collection;

Largest cells that are found in urine;

Flat straight edges, polygonal, angular cells with small round nucleus;

Prominent in estrus in females;

Usually not seen in samples collected by cystocentesis or catheterization;

Their presence is not considered signicant

Front 14

Transitional Epithelial Cells in Urine

Back 14

Large variation in size is normal;

From bladder, ureters, renal pelvis and part of urethra;

Small from kidneys and ureters;

Are usually round, pear shaped;

Granular cytoplasm, small nucleus;

Increased numbers in inflammation (cystitis) and are seen in catheterized samples

Front 15

Caudate Epithelial Cells in Urine

Back 15

From kidneys and ureters;

Have a cytoplasmic tail

Front 16

Renal Epithelial Cells in Urine

Back 16

Originate from renal tubules;

The smallest epithelials;

Slightly larger than WBCs;

Round and contain large nucleus;

Non-granular cytoplasm;

Seen in: renal disease and neoplaisa

Front 17

Bacteria

Back 17

Not normal in urine;

Proliferates;

Not detected until numbers reach 10,000/ml;

Refract light, rods or cocci;

Increased WBCs and bacteria indicate infections;

For best assessment do a stained cytologic preparation;

For further identification can do a culture and sensitivity or gram stain

Front 18

Casts

Back 18

Formed in the lumen of distal and collective tubules of the kidney;

Precipitate in acidic conditions;

Cylindrical with parallel sides;

Types include: hyaline, cellular, granular, waxy;

Dissolve in alkaline urine;

Indicate tubular damage (inflammation, irritation, degeneration);

Very fragile

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Hyaline Casts

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Few may be normal;

Clear, colourless and refractile, contain mucoproteins;

Rounded ends;

Increase indicates milk form of renal irritation;

Associated: strenuous exercise, fever, general anesthesia, poor renal perfusion, congestive heart failure

Front 20

Granular Casts

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Most common;

Hyaline casts containing granules;

Granules from epithelial and WBC incorporate in cast then degenerate;

Seen: acute nephritis, toxicity, severe kidney damage;

Describe as either coarse or fine

Front 21

Epithelial Casts

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Always includes renal epithelial cells because only kind in tubules at formation;

Granular casts usually accompany them;

Seen: Acute nephritis, and degeneration of epithelium`

Front 22

WBC Casts

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Contain and are surrounded with WBC, mainly neutrophils;

Inflammation of tubules;

Looks similar to renal epithelial casts;

Form granular casts as WBCs begin to degenerate

Front 23

RBC Casts

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Are yellow-orange in colour;

Indicate renal bleeding from hemorrhage in trauma or from inflammation lesions

Front 24

Waxy Casts

Back 24

Resemble hyaline casts but are wider with square ends;

Waxy appearance- more opaque;

Often see cracks in sides;

Indicate chronic, severe degeneration of renal tubules

Front 25

Fatty Casts

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Contain many small droplets of varying sizes;

Appear refractile;

Often seen in cats with renal disease;

Occasionally in dogs with diabetes;

Indicate degeneration of tubules

Front 26

Mucus Threads

Back 26

Resemble a twisted ribbon;

Large amount in horses because of mucus glands in the kidney and ureters;

In other species indicates urethral irritation or contamination with genital secretions

Front 27

Spermatozoa

Back 27

Seen in urine of intact males;

Have no clinical significance

Front 28

Fat Droplets

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Highly refractile spherical bodies of varying sizes;

Not same focus as other cells;

Contaminants from lubricants used in catheteriazation;

Seen in obesity, diabetes, hypothyroidism and high fat meals

Front 29

Fungi and Yeast

Back 29

Confused with lipid droplets;

Characteristic budding;

Unicellular;

Are contaminants in urine;

Fungi are branching filaments;

Are uncommon but serious when they happen

Front 30

Capillaria Plica

Back 30

Bladder worm of dogs and cats;

Have characteristic caps at both ends of the ova

Front 31

Dioctaphyma renale

Back 31

Kidney worms of dogs;

Have a wavy outer shell

Front 32

Stephanurus dentatus

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Kidney worms of pigs

Front 33

Crystalluria

Back 33

Consequence of elements secreted in urine by normal renal activity or metabolic diseases;

Type of crystals formed depend:

pH, temp, SpG, diet, and medication

Front 34

Acidic Urine

Back 34

Calcium oxalate;

Amorphous urates;

Sodium urates;

Uric acid;

Calcium sulfates;

Cystine

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Alkaline Urine

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Phosphate crystals or struvite;

Amorphous Phosphate;

Calcium carbonate;

Ammonium biurate

Front 36

Struvite Crystals

Back 36

Normally in alkaline urine, however can be found in slightly acidic urine;

Magnesium Ammonium phosphate, triple phosphate;

Are eight sided prisms- described as 'cofin lids';

Associated with urease producing bacteria of lower urinary tract disease;

Associated with uroliths;

Fern leaf shape in high ammonia concentrations;

Amorphous phosphate crystals appear as granular precipitate- are soluble in acetic acid so will dissolve in vinegar

Front 37

Amorphous Phosphate/Urate

Back 37

Phosphate found in alkaline urine and appear as granular precipitate;

Urates similar but found in acidic urine

Front 38

Calcium Carbonate

Back 38

Commonly seen in urine of horses, rabbits and guinea pigs;

Round with many lines radiating from center;

Also may have dumbbell shape

Front 39

Ammonium Biurate

Back 39

Are round and brown coloured with long spicules- thorn apple shape or mite appearance;

Common in animals with severe liver disease- portocaval shunts;

Dalmatians have uric acid metabolism predisposition- sometimes in English bulldogs;

Amorphous urates- granular like amorphous phosphates but in acidic urine;

In cats tend to be spherical

Front 40

Uric Acid Crystals

Back 40

Associated with dalmatians metabolic defects;

Flat square looking;

Common in humans

Front 41

Calcium Oxalate Dihydrate

Back 41

Appear as small squares containing an x inside;

Found in acidic and neutral urine;

Common in horses and cattle;

Min. Schnauzers prone to calcium oxalate uroliths;

Seen in calcium metabolism disorders;

Increased numbers in canine/felines indicate acute renal failure

Front 42

Calcium Oxalate Monohydrate

Back 42

Appear as elongated with pointed ends;

Seen in large numbers in ethylene glycol poisonings

Front 43

Tryosine and Leucine

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Tryosine are dark and needle like;

Leucine are small, round with sectioned centers;

Often found in clusters;

Seen in animals with liver disease

Front 44

Cystine

Back 44

Appear as six sided, hexagonal, flat plates;

Are indicative of metabolic disorders of tubules in kidneys;

Possible association with uroliths

Front 45

Bilirubin Crystals

Back 45

Often found in bilirubinuria;

Can indicate an underlying cholestatic process;

Yellowish-brown in color;

Most commonly appear as small, needle like crystals;

Cylindrical bilirubin crystals can form in association with droplets of fat resulting in a "flashlight" appearance;

'Bilirubin staining'

Front 46

Undiagnostic Crystals

Back 46

May form in standing urine;

Due to exposure of air resulting in moisture loss and crystal formation;

Can happen in refrigerated samples and room temperature samples;

Warming of the urine after refrigeration may cause these crystals to dissolve

Front 47

Artifacts

Back 47

Enter the sample collection and transportation;

Can be a source of confusion so recognition is important;

Air bubbles;

Oil droplets-from surgical gloves;

Hair;

Fecal material and possible parasite ova;

Dust;

Plant spores and pollens

Front 48

Urolithiasis

Back 48

Uroliths are calculi-stones composed of various minerals from urinary tract;

May block urethra or remain in bladder and cause inflammation and bleeding;

Determining composition important to determine therapy;

Must remove and prevent reoccurence;

Type of urolith can be determined by it's gross appearance, radiographic appearance and crystal types in sediment

Front 49

Triple Phosphate (Struvite) Uroliths

Back 49

Triple phosphate (struvite);

Most common in dogs;

Second most common in cats;

Radioopaque, hard, white, or yellow, powdery when crushed;

In alkaline urine

Front 50

Urate Uroliths

Back 50

Radiolucent, yellow brittle;

Common in Dalmatians

Front 51

Calcium Oxalate Uroliths

Back 51

Most common in cats;

Second most common in dogs;

Radioopaque, hard, not crush easily, have protrusions on surface;

Traumatizes bladder more

Front 52

Special Functional Tests

Back 52

These tests are performed on urine but serve to complement some chemistry tests;

Ex: urine protei/creatinine ratio;

Serves as a quantitive assessment for proteinuria;

To verify glomerular disease;

Based on fact that tubular concentration of urine increases both protein and creatinine concentrations equally in normal circumstances;

5 mls collected between 10 am and 2 pm by cystocentesis;

Sample is kept cool-usually sent to reference lab or centrifuged and supernatant is used in a commercial testing kit;

Normal P/C ratio is less than 1;

1-5 indicates prerenal or functional origin;

>5 indicates renal disease