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URINALYSIS
OVERVIEW


1. Urinalysis

2. Serum/plasma urea & creatinine concentrations

3. Urine protein to creatinine ratio (UPC ratio)



                                                   2
1.URINALYSIS
MAIN INDICATIONS ARE:
  • Evaluation of renal & lower urinary tract abnormalities
  • Assessment of some metabolic/endocrine disorders
  • Assessment of state of hydration




                                                              3
Gross evaluation


             Specific gravity (SG)
Urinalysis
             Biochemical analysis


             Sediment examination
                 (microscopy)

                   Culture           4
GROSS EVALUATION: COLOUR




Urine from healthy animals can vary in colour                   Red discolouration
but is usually light, mild or dark yellow


  Red discolouration may indicate haemoglobinuria, myoglobinuria or haematuria.   5
GROSS EVALUATION: TURBIDITY




                                              turbidity in urine from
Upper sample :Clear urine   Turbid urine           a cow with
Lower sample:Turbid         (healthy horse)      pyelonephritis 6
TURBIDITY

 Healthy horses and rabbits may have turbid
 urine due to high concentration of mucin and
 crystals. In other species turbidity can indicate
 the presence of sediment.
 On refrigeration, urine samples may become
 turbid from crystallisation of minerals which
 were in solution, and they may clear when
 returned to room temperature

                                                     7
Gross evaluation


             Specific gravity (SG)
Urinalysis
             Biochemical analysis


             Sediment examination
                 (microscopy)

                   Culture           8
SPECIFIC GRAVITY (SG)
Ratio of weight (density) of urine to that of an equal volume of water
at the same temperature. No units.

     Values depend on:

              - hydration status and water intake

              - the kidney’s concentrating ability

     It is a test of renal tubular function


   Hydration status can be determined by assessing skin turgor or by
   measurement of serum albumin, or PCV and total proteins.            9
SPECIFIC GRAVITY (SG): MEASUREMENT
–   Reagent test strips are unreliable for animals/Always use
    the refractometer
–   If the urine is turbid, centrifuge it before measuring SG of
    the supernatant


                   1.050
                   1.040
                   1.030
                   1.020
                   1.010
                   1.000
                                                             10
SG: INTERPRETATION
HYPERSTHENURIA: concentrated urine
  - >1.012
  -urine of healthy, normally hydrated animals

ISOSTHENURIA: urine neither concentrated nor diluted
  -1.007-1.012 (urine SG = plasma filtrate SG)
  -persistent isosthenuria warrants further investigation

HYPOSTHENURIA: urine is more diluted than plasma
 - <1.007
- persistent hyposthenuria warrants further investigation
                                                        11
SG: INTERPRETATION
 *   The range of values for SG can vary according to water intake and
     hydration status. Usually SG in normal concentrated urine is >1.030
 *   WATER DEPRIVATION TEST
 *   Contra-indication: It should never be carried out in depressed,
     dehydrated or azotemic animals, or if renal failure is suspected.
 *   Indication: Confirmation of the animals ability to concentrate its urine
     when water is withheld.
 *   Protocol: The urine SG is monitored every 2 hours until 5% of body
     weight is lost, or the urine SG is >1.020.
 *   Interpretation:
      *   If the urine SG increases to 1.020, tubular function and ADH availability are
          confirmed.
      *   If the urine SG remains <1.020, diabetes insipidus is suspected.


                                                                                          12
Gross evaluation


             Specific gravity (SG)
Urinalysis
             Biochemical analysis


             Sediment examination
                 (microscopy)

                   Culture           13
BIOCHEMICAL ANALYSIS URINE STRIPS




   Always follow manufacturer instructions
                                             14
EXAMPLE OF A URINE STRIP CHART




                                 15
GLUCOSE
- Glucose is not normally found in urine of healthy animals
- Causes of glycosuria
  • Persistent hyperglycaemia
     - Diabetes mellitus
  • Transient hyperglycaemia
     - Stress in cats
     - Drugs (xylazine, ketamine)
     - IV fluids containing glucose
     - Convulsions
  • Renal tubular disorders
     - Fanconi syndrome
     - Primary glucosuria                                16
BILIRUBIN
–   Not accurate for dogs/cats
    • Tests utilising a tablet (ictotest) can be more accurate than strip-
       tests
    • Light can break down bilirubin
–   Trace to + normal in healthy dogs.No bilirubin present in
    the urine of other healthy animals
–   The bilirubin in the urine is water-soluble conjugated
    bilirubin
–   Causes of bilirubinuria
     • Same as causes of bilirubinaemia

                                                                       17
KETONES

  –   Accurate test for animals
       • Does not detect β-hydroxybutyric acid

  –   Ketones are not present in the urine of clinically healthy
      animals
       • Trace can be normal in rabbits
  –   Causes of ketonuria
       • Diabetes mellitus, pregnancy, starvation, ketosis, immediately after
         calving in high–producing dairy cows
N.B. Many disorders causing anorexia in cattle (e.g. mastitis, metritis, pneumonia)
will cause ketonaemia and ketonuria, but levels of ketones are generally not as high
as in primary ketosis.
                                                                                       18
BLOOD / HAEMOGLOBIN


- Accurate test for animals

- Detects intact RBCs, haemoglobin or myoglobin

-   Follow-up positive result with sediment examination
-   Interpret positive result in conjunction with the method
    of urine collection (cystocentesis can be a cause of presence of
    blood in urine)


                                                                       19
pH
– Acceptable test for animals
- Carnivores:

      • acidic urine is normal if fed a meat diet
      • alkaline urine usually reflects urinary tract infection
- Herbivores:
      • alkaline urine is normal
      • acidic urine may reflect increased protein catabolism
             e.g. high protein diet, starvation, fever, nursing animals
–   Some drugs can influence pH
–   Not an accurate indicator of systemic acid/base balance
                                                                          20
PROTEIN

–   Acceptable test for animals but can give false positive reaction in
    alkaline samples.
–   Test detects mainly albumin. Does not detect globulins
–   Always interpret in conjunction with SG and sediment examination
    (it is not abnormal to have trace protein in concentrated urine but
    always abnormal finding in diluted urine).
–   Common causes of proteinuria:
       •   urogenital haemorrhage
       •   urogenital inflammation
       •   renal protein loss

                                                                          21
NITRITE, UROBILINOGEN, LEUKOCYTES

- Nitrite
   • Positive results may indicate bacterial infection
   • false negative results occur commonly

- Urobilinogen
   • Questionable clinical usefulness

- Leukocytes
   • False negative results common in dogs
   • False positive results common in cats



   NONE ARE RELIABLE IN EXAMINING ANIMAL URINE
                                                         22
2. Serum urea & creatinine
      concentrations




                             23
UREA
                          LIVER
                                               BLOOD
                           Urea cycle
                                               Urea
    TISSUES                             Urea
                          NH4+
                                               75%

  proteins         NH4+                                KIDNEYS

                                  25%
                                               Urea
    Urea
Dietary Proteins
    bacteria              NH4+
                                                        Urea in
    GASTROINTESTINAL TRACT                              urine
                                                            24
UREA and creatinine

–   Glomeruli: 75% of urea is excreted (excretion ↑ or ↓
    when glomerular filtration rate ↑ or ↓ )
–   Tubules: Urea is reabsorbed (reabsorption ↑ or ↓ when
    glomerular filtration rate ↓ or ↑)
–   Creatinine is derived from creatine-phosphate,
    creatinine is excreted via the glomeruli. It is not
    reabsorbed in the tubules so excretion of creatinine is
    a measure of glomerular filtration rate.

                                                          25
BUN vs. UREA

–   BUN = blood urea nitrogen= concentration of the nitrogen
    component of urea in blood

–   BUN value is Lower than urea value. BUN:Urea ratio is
    approximately 1:2

–   But the term BUN is used interchangeably with urea


                                                            26
CREATININE
                                         KIDNEYS

  creatine     creatinine                creatinine

MUSCLE

             creatinine
  BLOOD
                                           Creatinine in
                                           urine


                     creatinine   NH4+
    INTESTINES
                                                      27
AZOTAEMIA

- Increased serum/plasma urea & creatinine
  concentrations


                    URAEMIA

– Marked azotaemia and clinical signs (vomiting,
  anorexia, gastrointestinal ulceration)


                                                   28
AZOTAEMIA


CAUSES

 PRE-RENAL

 RENAL

 POST-RENAL



              29
PRE-RENAL AZOTAEMIA
- DECREASED RENAL PERFUSION
 - Hypovolaemia, dehydration, cardiovascular disease
       Urea is ↑ and creatinine Normal / ↑
       Urine specific gravity is ↑
- INCREASED UREA PRODUCTION
    - G.I. TRACT HAEMORRHAGE
       Urea is ↑ and creatinine is Normal
    - HIGH PROTEIN DIET
       Urea is ↑ and creatinine is Normal
                                                       30
RENAL AZOTAEMIA



- RENAL DISEASE
  - ONLY evident when more than 60-75% of nephrons
    are compromised


  •   Urea and creatinine are ↑
  •   Urine is inadequately concentrated


                                                     31
POST-RENAL AZOTAEMIA

- URINARY TRACT OBSTRUCTION
 • ureter, urethra

- URINARY TRACT RUPTURE
 • ureter, bladder, urethra



 - Urea and creatinine are ↑

                               32
LOW SERUM/PLASMA UREA: CAUSES

–   Decreased Liver Function
–   Portosystemic shunt
–   Increased Excretion
     • Extreme PU/PD
     • Overhydration
–   Low protein intake

Young Animals have a lower reference range


                                             33
RUMINANTS / HORSES


- Excrete most of urea via the gut (very little via kidneys). So

  blood urea can be normal despite severe renal disease.

- Therefore blood creatinine is a more sensitive indicator of

  renal disease



                                                             34
3.Urine protein to urine
creatinine ratio (UPC ratio)




                               35
UPC RATIO
–   Used to assess the clinical significance of proteinuria
–   Total protein & creatinine concentrations are measured in a single
    urine sample and expressed in the same units

        UPC RATIO               INTERPRETATION
        < 1.0                   Proteinuria if present is not significant

        MILD/MODERATE           Significant proteinuria from:
        INCREASES (1-2)             urogenital haemorrhage
                                    urogenital inflammation
                                    glomerular protein loss

        MODERATE/SEVERE Usually glomerular protein loss
        INCREASES (>2)
                                                                            36
Urine Sediment Examination
 Produced by centrifugation of urine onto a slide
 Looking for:
     Abnormal     crystals
     Cells
     Infectious   agents
     Casts
     Contaminants


                                                     37
Ammonium biurate: suggestive of liver dysfunction and/or portosystemic shunting




                                                                                  38
Conjugated bilirubin: normal in dogs, but abnormal in other species. Suggests
conjugated bilirubinemia



                                                                                39
Calcium oxalate: suggestive of hypercalciuria, hyperoxaluria (dietary, ethylene
glycol) or calcium oxalate uroliths

Can occur normally if urine has been stored

                                                                                  40
Magnesium ammonium phosphate (struvite) – present in normal animals but
excessive amounts can be an indicator of struvite uroliths or mixed uroliths

Common in bacterial-induced alkalinuria

                                                                               41
Calcium oxalate monohydrate – suggestive of hypercalciuria or EG toxicity




                                                                            42
Cysteine – indicative of disruption of cysteine metabolism/catabolism (drug induced
or inborn error in metabolism



                                                                                      43
Drug-associated - sulfonamides




                                 44
Tyrosine – associated with severe liver disease




                                                  45
Tyrosine – associated with severe liver disease




                                                  46
RBCs – up to 5 RBC per high power field is normal. Increased RBCs in urine
generally indicates hemorrhage into the bladder or urethra (artifact with catheter
collection in some cases)


                                                                                     47
WBCs – up to 5 WBC per high power field is normal, higher levels suggest infection
somewhere within the urinary tract



                                                                                     48
Transitional epithelial cells from renal pelvis, ureters, urinary bladder or urethra

Morphology used for detection of neoplastic change (cytology)


                                                                                       49
Squamous epithelial cells – indicator of contamination from skin, prepuce etc




                                                                                50
Neoplastic cells – commonly transitional cell carcinomas




                                                           51
Bacteria – bacili on the right, cocci on the left. Presence is always abnormal – either
infection or contamination (storage, collection)



                                                                                          52
Fungi – presence is always abnormal – Candidiasis or Aspergillus sp. In the case of
infection. Be aware: contamination, storage, growth on microscope slides or in stain
preparations
                                                                                       53
Parasites




            54
Casts:

Hyaline casts = precipitated protein

Cellular casts: These have distinct cells within
the protein matrix - if the cells are of epithelial
origin (i.e., not WBCs or RBCs), they are called
epithelial casts..

Granular casts: As cells within the protein cast
matrix break down, the cast becomes coarsely
then finely granular.

Waxy casts: Waxy casts are the final stage of
cast degeneration (usually originating from
cellular and granular casts). Compared to
hyaline casts, they are readily observable
because they have a smooth appearance, no
internal texture, and are more refractile than the
surrounding urine.

                                             55
56
57
Questions?




             58

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Urinalysis

  • 2. OVERVIEW 1. Urinalysis 2. Serum/plasma urea & creatinine concentrations 3. Urine protein to creatinine ratio (UPC ratio) 2
  • 3. 1.URINALYSIS MAIN INDICATIONS ARE: • Evaluation of renal & lower urinary tract abnormalities • Assessment of some metabolic/endocrine disorders • Assessment of state of hydration 3
  • 4. Gross evaluation Specific gravity (SG) Urinalysis Biochemical analysis Sediment examination (microscopy) Culture 4
  • 5. GROSS EVALUATION: COLOUR Urine from healthy animals can vary in colour Red discolouration but is usually light, mild or dark yellow Red discolouration may indicate haemoglobinuria, myoglobinuria or haematuria. 5
  • 6. GROSS EVALUATION: TURBIDITY turbidity in urine from Upper sample :Clear urine Turbid urine a cow with Lower sample:Turbid (healthy horse) pyelonephritis 6
  • 7. TURBIDITY  Healthy horses and rabbits may have turbid urine due to high concentration of mucin and crystals. In other species turbidity can indicate the presence of sediment.  On refrigeration, urine samples may become turbid from crystallisation of minerals which were in solution, and they may clear when returned to room temperature 7
  • 8. Gross evaluation Specific gravity (SG) Urinalysis Biochemical analysis Sediment examination (microscopy) Culture 8
  • 9. SPECIFIC GRAVITY (SG) Ratio of weight (density) of urine to that of an equal volume of water at the same temperature. No units. Values depend on: - hydration status and water intake - the kidney’s concentrating ability It is a test of renal tubular function Hydration status can be determined by assessing skin turgor or by measurement of serum albumin, or PCV and total proteins. 9
  • 10. SPECIFIC GRAVITY (SG): MEASUREMENT – Reagent test strips are unreliable for animals/Always use the refractometer – If the urine is turbid, centrifuge it before measuring SG of the supernatant 1.050 1.040 1.030 1.020 1.010 1.000 10
  • 11. SG: INTERPRETATION HYPERSTHENURIA: concentrated urine - >1.012 -urine of healthy, normally hydrated animals ISOSTHENURIA: urine neither concentrated nor diluted -1.007-1.012 (urine SG = plasma filtrate SG) -persistent isosthenuria warrants further investigation HYPOSTHENURIA: urine is more diluted than plasma - <1.007 - persistent hyposthenuria warrants further investigation 11
  • 12. SG: INTERPRETATION * The range of values for SG can vary according to water intake and hydration status. Usually SG in normal concentrated urine is >1.030 * WATER DEPRIVATION TEST * Contra-indication: It should never be carried out in depressed, dehydrated or azotemic animals, or if renal failure is suspected. * Indication: Confirmation of the animals ability to concentrate its urine when water is withheld. * Protocol: The urine SG is monitored every 2 hours until 5% of body weight is lost, or the urine SG is >1.020. * Interpretation: * If the urine SG increases to 1.020, tubular function and ADH availability are confirmed. * If the urine SG remains <1.020, diabetes insipidus is suspected. 12
  • 13. Gross evaluation Specific gravity (SG) Urinalysis Biochemical analysis Sediment examination (microscopy) Culture 13
  • 14. BIOCHEMICAL ANALYSIS URINE STRIPS Always follow manufacturer instructions 14
  • 15. EXAMPLE OF A URINE STRIP CHART 15
  • 16. GLUCOSE - Glucose is not normally found in urine of healthy animals - Causes of glycosuria • Persistent hyperglycaemia - Diabetes mellitus • Transient hyperglycaemia - Stress in cats - Drugs (xylazine, ketamine) - IV fluids containing glucose - Convulsions • Renal tubular disorders - Fanconi syndrome - Primary glucosuria 16
  • 17. BILIRUBIN – Not accurate for dogs/cats • Tests utilising a tablet (ictotest) can be more accurate than strip- tests • Light can break down bilirubin – Trace to + normal in healthy dogs.No bilirubin present in the urine of other healthy animals – The bilirubin in the urine is water-soluble conjugated bilirubin – Causes of bilirubinuria • Same as causes of bilirubinaemia 17
  • 18. KETONES – Accurate test for animals • Does not detect β-hydroxybutyric acid – Ketones are not present in the urine of clinically healthy animals • Trace can be normal in rabbits – Causes of ketonuria • Diabetes mellitus, pregnancy, starvation, ketosis, immediately after calving in high–producing dairy cows N.B. Many disorders causing anorexia in cattle (e.g. mastitis, metritis, pneumonia) will cause ketonaemia and ketonuria, but levels of ketones are generally not as high as in primary ketosis. 18
  • 19. BLOOD / HAEMOGLOBIN - Accurate test for animals - Detects intact RBCs, haemoglobin or myoglobin - Follow-up positive result with sediment examination - Interpret positive result in conjunction with the method of urine collection (cystocentesis can be a cause of presence of blood in urine) 19
  • 20. pH – Acceptable test for animals - Carnivores: • acidic urine is normal if fed a meat diet • alkaline urine usually reflects urinary tract infection - Herbivores: • alkaline urine is normal • acidic urine may reflect increased protein catabolism e.g. high protein diet, starvation, fever, nursing animals – Some drugs can influence pH – Not an accurate indicator of systemic acid/base balance 20
  • 21. PROTEIN – Acceptable test for animals but can give false positive reaction in alkaline samples. – Test detects mainly albumin. Does not detect globulins – Always interpret in conjunction with SG and sediment examination (it is not abnormal to have trace protein in concentrated urine but always abnormal finding in diluted urine). – Common causes of proteinuria: • urogenital haemorrhage • urogenital inflammation • renal protein loss 21
  • 22. NITRITE, UROBILINOGEN, LEUKOCYTES - Nitrite • Positive results may indicate bacterial infection • false negative results occur commonly - Urobilinogen • Questionable clinical usefulness - Leukocytes • False negative results common in dogs • False positive results common in cats NONE ARE RELIABLE IN EXAMINING ANIMAL URINE 22
  • 23. 2. Serum urea & creatinine concentrations 23
  • 24. UREA LIVER BLOOD Urea cycle Urea TISSUES Urea NH4+ 75% proteins NH4+ KIDNEYS 25% Urea Urea Dietary Proteins bacteria NH4+ Urea in GASTROINTESTINAL TRACT urine 24
  • 25. UREA and creatinine – Glomeruli: 75% of urea is excreted (excretion ↑ or ↓ when glomerular filtration rate ↑ or ↓ ) – Tubules: Urea is reabsorbed (reabsorption ↑ or ↓ when glomerular filtration rate ↓ or ↑) – Creatinine is derived from creatine-phosphate, creatinine is excreted via the glomeruli. It is not reabsorbed in the tubules so excretion of creatinine is a measure of glomerular filtration rate. 25
  • 26. BUN vs. UREA – BUN = blood urea nitrogen= concentration of the nitrogen component of urea in blood – BUN value is Lower than urea value. BUN:Urea ratio is approximately 1:2 – But the term BUN is used interchangeably with urea 26
  • 27. CREATININE KIDNEYS creatine creatinine creatinine MUSCLE creatinine BLOOD Creatinine in urine creatinine NH4+ INTESTINES 27
  • 28. AZOTAEMIA - Increased serum/plasma urea & creatinine concentrations URAEMIA – Marked azotaemia and clinical signs (vomiting, anorexia, gastrointestinal ulceration) 28
  • 30. PRE-RENAL AZOTAEMIA - DECREASED RENAL PERFUSION - Hypovolaemia, dehydration, cardiovascular disease  Urea is ↑ and creatinine Normal / ↑  Urine specific gravity is ↑ - INCREASED UREA PRODUCTION - G.I. TRACT HAEMORRHAGE  Urea is ↑ and creatinine is Normal - HIGH PROTEIN DIET  Urea is ↑ and creatinine is Normal 30
  • 31. RENAL AZOTAEMIA - RENAL DISEASE - ONLY evident when more than 60-75% of nephrons are compromised • Urea and creatinine are ↑ • Urine is inadequately concentrated 31
  • 32. POST-RENAL AZOTAEMIA - URINARY TRACT OBSTRUCTION • ureter, urethra - URINARY TRACT RUPTURE • ureter, bladder, urethra - Urea and creatinine are ↑ 32
  • 33. LOW SERUM/PLASMA UREA: CAUSES – Decreased Liver Function – Portosystemic shunt – Increased Excretion • Extreme PU/PD • Overhydration – Low protein intake Young Animals have a lower reference range 33
  • 34. RUMINANTS / HORSES - Excrete most of urea via the gut (very little via kidneys). So blood urea can be normal despite severe renal disease. - Therefore blood creatinine is a more sensitive indicator of renal disease 34
  • 35. 3.Urine protein to urine creatinine ratio (UPC ratio) 35
  • 36. UPC RATIO – Used to assess the clinical significance of proteinuria – Total protein & creatinine concentrations are measured in a single urine sample and expressed in the same units UPC RATIO INTERPRETATION < 1.0 Proteinuria if present is not significant MILD/MODERATE Significant proteinuria from: INCREASES (1-2) urogenital haemorrhage urogenital inflammation glomerular protein loss MODERATE/SEVERE Usually glomerular protein loss INCREASES (>2) 36
  • 37. Urine Sediment Examination  Produced by centrifugation of urine onto a slide  Looking for:  Abnormal crystals  Cells  Infectious agents  Casts  Contaminants 37
  • 38. Ammonium biurate: suggestive of liver dysfunction and/or portosystemic shunting 38
  • 39. Conjugated bilirubin: normal in dogs, but abnormal in other species. Suggests conjugated bilirubinemia 39
  • 40. Calcium oxalate: suggestive of hypercalciuria, hyperoxaluria (dietary, ethylene glycol) or calcium oxalate uroliths Can occur normally if urine has been stored 40
  • 41. Magnesium ammonium phosphate (struvite) – present in normal animals but excessive amounts can be an indicator of struvite uroliths or mixed uroliths Common in bacterial-induced alkalinuria 41
  • 42. Calcium oxalate monohydrate – suggestive of hypercalciuria or EG toxicity 42
  • 43. Cysteine – indicative of disruption of cysteine metabolism/catabolism (drug induced or inborn error in metabolism 43
  • 45. Tyrosine – associated with severe liver disease 45
  • 46. Tyrosine – associated with severe liver disease 46
  • 47. RBCs – up to 5 RBC per high power field is normal. Increased RBCs in urine generally indicates hemorrhage into the bladder or urethra (artifact with catheter collection in some cases) 47
  • 48. WBCs – up to 5 WBC per high power field is normal, higher levels suggest infection somewhere within the urinary tract 48
  • 49. Transitional epithelial cells from renal pelvis, ureters, urinary bladder or urethra Morphology used for detection of neoplastic change (cytology) 49
  • 50. Squamous epithelial cells – indicator of contamination from skin, prepuce etc 50
  • 51. Neoplastic cells – commonly transitional cell carcinomas 51
  • 52. Bacteria – bacili on the right, cocci on the left. Presence is always abnormal – either infection or contamination (storage, collection) 52
  • 53. Fungi – presence is always abnormal – Candidiasis or Aspergillus sp. In the case of infection. Be aware: contamination, storage, growth on microscope slides or in stain preparations 53
  • 54. Parasites 54
  • 55. Casts: Hyaline casts = precipitated protein Cellular casts: These have distinct cells within the protein matrix - if the cells are of epithelial origin (i.e., not WBCs or RBCs), they are called epithelial casts.. Granular casts: As cells within the protein cast matrix break down, the cast becomes coarsely then finely granular. Waxy casts: Waxy casts are the final stage of cast degeneration (usually originating from cellular and granular casts). Compared to hyaline casts, they are readily observable because they have a smooth appearance, no internal texture, and are more refractile than the surrounding urine. 55
  • 56. 56
  • 57. 57