Recommended testing and sample requirements:

Test	Tube	Sample Volume	Components
Haematology	Lithium Heparin (preferred) or EDTA+ 2x air-dried smears	Minimum 300 μL	Packed cell volume Estimated white cell count White cell differential Morphology of red and white cells Thrombocyte assessment
Biochemistry*	Lithium Heparin Plain (also accepted)	Minimum 400 μL or 300 μL (no electrolytes)]	Liver disease Renal disease General	AST GLDH Bile acids Uric acid Total protein Glucose Calcium Phosphorus CK

* For small volume samples please indicate biochemistry analytes in order of priority.

Interpretation of Biochemistry

 As interpretation of avian chemistry results differs from mammalian chemistry, a brief explanation of the various tests is included as follows:

 Liver and Muscle

1. AST and Bile Acids are the most sensitive indicators of liver disease in birds.

• AST is not liver specific and can be increased in any septic or inflammatory condition, muscle disease and with certain antibiotics and steroids.
• GLDH is present within hepatocyte mitochondria and is considered the most specific indicator for hepatocellular damage in birds. 
  • GLDH also has high activity in renal tissue in birds but most of the enzyme is excreted directly into urine and never reaches the blood.
•  Serum bile acid concentration is a reliable indicator of liver function.  
  •  Requires serum sample as heparin interferes with the assay.
  • Feeding can increase bile acids up to 1.6-4.5 x the normal reference range therefore fasting samples are preferred – pigeons, ostriches and some parrots lack a gall bladder so fasting is not needed in these species.
  • Amazon parrots normally have slightly higher bile acids than other companion avian species
  • Low bile acid concentrations are common in birds with microhepatica, poor feather formation, and an overgrown malformed beak.

Generally, a bile acid concentration > 100 μmol/L is considered abnormal and > 75μmol/L is suspicious for hepatic insufficiency.

2. CK activity increases with muscle damage and this, along with AST and GLDH, is used to differentiate muscle from liver disease.  

3. The following tests are NOT recommended as indicators of liver disease in birds:

• ALT and GGT occurs in many different tissues and in some species enzyme activity is below the sensitivity of many analysers.
• ALP activity can be increased due to inflammation therefore is considered nonspecific.
• Bilirubin concentrations increase inconsistently in cases of liver disease and some birds (chickens) cannot form bilirubin

 Renal function

1. Uric Acid is the most reliable test of renal disease.

• It requires more serum or plasma than the other tests (a minimum of 50μL)so is often done last.
• Age, diet, sex, and recent feeding may affect results especially in raptors where ppstprandial levels can increase to twice normal for up to 8 hours.
• Elevations are seen with severe dehydration and renal disease.
• It can be used as aprognostic indicator for gout – the solubility of uric acid in plasma in birds is around 600 μmol/L – levels higher than this (1500– 2500μml/L) will lead to precipitation in joints.

2.  Urea and creatinine are not useful tests for monitoring for renal disease.

• Urea can be useful in assessing hydration status – concentrations of 0.4 – 0.7 mmol/L are considered normal, but up to 10-15x increase can be seen in dehydration.
• Creatinine is not synthesised by birds – most muscle breakdown products are excreted as creatine rather than creatinine.

Other Tests

1. Protein concentrations in serum are generally lower in birds than mammals.

• In most avian species levels range between 20-40 g/L (some spp. are as low as 15 g/L)
• Plasma concentrations will be approximately 1-2g/l higher than serum concentrations due to the presence of fibrinogen.

2. Glucose is not utilised by avian erythrocytes therefore concentrations in serum are much more stable in birds.  

• Levels will fall slowly over 24-48 hours if the serum stays on the clot.
• If there is a delay in the sample reaching the laboratory, either spin and separate the sample or use a fluoride oxalate (grey top) tube.

The glucose reference range in birds (11.2-27.7 mmol/L) is generally higher than in mammals

• Stress and postprandial levels can cause transient increases to 24.9 – 33.3 mmol/L.
• Conditions such as egg yolk peritonitis and renal carcinoma can also cause similar increases
• Borderline  hypoglycaemia is common in cockatoos and probably of no clinical significance

3. Calcium measured in serum includes albumin bound and free forms therefore concentration varies with albumin concentration.

• Oestrogen induces hypercalcaemia therefore calcium concentration in serum increases approximately 4 days prior to ovulation.
• Corticosteroids decrease total calcium.
• As in mammals dehydration and some tumors can increase calcium concentrations.
• African Grey Parrots have a described idiopathic hypocalcaemia (a unique form of hypoparathyroidism in which calcium is not properly released from bone).

 Avian Urinalysis

•  Urine collection can be aided by giving water by a crop tube – the bird will often urinate shortly after.
• Avian urine has a concentrated white to off white uric acid component and clear watery component – the watery fraction is assessed.

pH	Ranges from 6.0-8.0 (diet related)
Protein	Trace amounts normal
Glucose	Trace amounts normal in some species. Will occur if blood glucose is >33.3 mmol/L.
Ketones	Negative in normal birds. Increased with diabetes & increased fat metabolism
Specific gravity	1.005-1.020
Bilirubin	Not normally present. Biliverdin is the major bile pigment which does not react with the mammalian urine dip stick
Blood	Negative or trace
Nitrate	Unreliable in birds
Urates	White, pale yellow and pale grey are normal
Liver disease	Urates are yellow to green
Acute lead poisoning	Urates brown or blood tinged
Sediment*	RBC & WBC: normal <3/HPF Epithelial cells: Normally none present Casts: Presence is associated with renal disease Bacteria: False positive with faecal contamination of urine (can proliferate in transit)

 * A fresh or recently refrigerated sample is required. Prolonged storage causes lysis of cells.