Preliminary Evaluation of Serum Protein Electrophoresis as a Diagnostic Tool in the Black Rhinoceros (Diceros bicornis)
Abstract
Serum protein electrophoresis (SPE) is a valuable tool for the diagnosis of certain diseases in humans and animals.1,3-8,12 This technique could be particularly beneficial when applied to health monitoring in the black rhinoceros (Diceros bicornis), a species which is predisposed to a number of diseases in captivity, the etiologies of which are not yet fully understood.2,14-19 Serum protein fractionation varies widely between species, thus it is necessary to establish a reference range in order to interpret SPE for a given species.10,13,20 This study has two purposes. The first is to determine normal ranges and patterns of serum proteins separated by SPE in the black rhinoceros. The second is to make a preliminary evaluation of the diagnostic potential of SPE for this species.
Serum protein electrophoresis was performed on samples from both clinically healthy (n=38) and clinically ill (n=16) black rhinoceroses. Data gathered from healthy individuals were used to establish reference ranges for SPE. The absolute ranges for total protein, albumin, and gamma (γ) globulins in the black rhinoceros are similar to those of domestic mammals, however, the albumin to globulin ratio and alpha (α) globulins tend to be lower while the beta (β) globulins tend to be higher in the black rhinoceros than in domestic mammals. Preliminary evaluation of data gathered from ill and subclinically ill individuals indicates that serum protein electrophoresis is helpful in the diagnosis of clinically inapparent disease in the black rhinoceros. Research toward identifying and characterizing electrophoretic patterns associated with subclinical and clinical disease in the black rhinoceros continues.
Table 1. Serum protein electrophoresis: protein fractions and components9,11
Protein fraction
|
Major protein constituents
|
Albumin
|
Albumin
|
|
α1 Globulins
|
α1-Acid glycoprotein α1-Antichymotrypsin
|
α1-Antithrombin III α1-Antitrypsin
|
α1-Lipoproteina
|
α2 Globulins
|
α2-Globulin α2-Lipoproteinb
|
α2-Macroglobulin Ceruloplasmin
|
Haptoglobin Protein C
|
β Globulins
|
Amyloid A β2-Lipoproteinc C3 Complement Transferrin
|
C4 Complement C-Reactive protein Ferritin
|
Hemopexin Plasminogen
|
γ Globulins
|
Immunoglobin A Immunoglobin E
|
Immunoglobin G Immunoglobin M
|
|
aHigh density lipoprotein (HDL)
bVery low density lipoprotein (VLDL)
cLow density lipoprotein (LDL)
Table 2. Differential diagnoses of serum protein alterations9,11
Protein fraction
|
Differential diagnosesa
|
Albumin
|
Increase
|
Dehydration
|
|
Decrease
|
Hepatic disease, gastrointestinal disease, renal disease, internal parasites, overhydration, malnutrition, blood or plasma loss
|
α1 Globulins
|
Increase
|
Acute inflammatory disease, pregnancy
|
|
Decrease
|
Hepatic disease, pulmonary disease, nephrotic syndrome
|
α2 Globulins
|
Increase
|
Acute inflammatory disease, nephrotic syndrome, hepatic disease, diabetes mellitus, hypothyroidism
|
β Globulins
|
Increase
|
Acute hepatitis, chronic active hepatitis, nephrotic syndrome, suppurative dermatopathy, anemias
|
|
Decrease
|
Autoimmune disease
|
γ Globulins
|
Increase
|
Chronic inflammatory disease, immune mediated disease, infectious disease, suppurative disease, connective tissue disease, multiple myeloma, lymphosarcoma
|
|
Decrease
|
Immune deficiency diseases
|
aEvaluation of serum protein status should include assessment of the albumin to globulin ratio (A:G)
Table 3. Reference ranges for SPE in the black rhinoceros (n=33)
|
Quantity (g/dl)
|
Percent of total protein (%)
|
Protein fraction
|
Range
|
Mean
|
SD
|
Range
|
Mean
|
SD
|
Total protein
|
6.00–8.40
|
7.37
|
0.71
|
N/A
|
N/A
|
N/A
|
A:G ratio
|
0.31–0.79
|
0.53
|
0.13
|
N/A
|
N/A
|
N/A
|
α Globulins
|
1.79–3.41
|
2.49
|
0.37
|
23.6–44.1
|
33.5
|
5.20
|
α1 Globulins
|
0.09–0.27
|
0.16
|
0.05
|
1.2–4.7
|
2.3
|
0.84
|
α2 Globulins
|
0.23–0.68
|
0.45
|
0.11
|
3.9–9.2
|
6.0
|
1.31
|
β Globulins
|
1.69–3.33
|
2.51
|
0.38
|
26.9–43.8
|
34.3
|
5.05
|
γ Globulins
|
1.08–2.58
|
1.76
|
0.43
|
15.4–34.4
|
23.8
|
4.58
|
Table 4. SPE reference ranges (g/dl) for the black rhinoceros compared to domestic animals
Protein fraction
|
Black rhinocerosa,b
|
Horsec
|
Cowc
|
Dogc
|
Catc
|
Total protein
|
6.00–8.40
|
5.20–7.90
|
6.74–7.46
|
5.40–7.10
|
5.40–7.80
|
A:G ratio
|
0.31–0.78
|
0.62–1.46
|
0.84–0.94
|
0.59–1.11
|
0.45–1.19
|
Albumin
|
1.79–3.02
|
2.60–3.70
|
3.03–3.55
|
2.60–3.30
|
2.10–3.30
|
α Globulins
|
0.33–1.00
|
0.37–2.01
|
0.75–0.88
|
0.50–1.60
|
0.50–1.60
|
α1 Globulins
|
0.10–0.32
|
0.06–0.70
|
|
0.20–0.50
|
0.20–0.50
|
α2 Globulins
|
0.23–0.68
|
0.31–1.31
|
|
0.30–1.10
|
0.30–1.10
|
β Globulins
|
1.81–3.33
|
0.69–2.47
|
0.80–1.12
|
1.30–2.70
|
1.30–2.70
|
γ Globulins
|
1.08–2.58
|
0.55–1.90
|
1.69–2.25
|
0.90–2.20
|
1.70–4.40
|
aReference ranges for the black rhinoceros are repeated from Table 4 for ease of comparison
bRhinoceros values determined by agarose gel SPE
cDomestic animal values by cellulose acetate SPE9
Acknowledgments
The authors would like to thank Howard Gilman, John Lukas, Dave Thompson, Cyd Teare, and the staff of White Oak Conservation Center; and Carolyn Cray from the University of Miami, Division of Comparative Pathology, for making this research possible. The authors also thank Eric Miller and the staff of the Saint Louis Zoo; Dave Jessup of the California Department of Fish and Game; Robin Radcliffe and the staff of Fossil Rim Wildlife Center; Mitch Finnegan and the staff of the Oregon Zoo; Tom Alvarado and the staff of the Dallas Zoo; Chris Miller and the staff of the Miami Metro Zoo; Peregrine Wolff, Michelle Miller, and the staff of Disney’s Animal Kingdom; Nancy Lung and the staff of the Fort Worth Zoo; Ray Ball and the staff of Busch Gardens, Tampa; Nadine Lamberski and the staff of Riverbanks Zoological Park; and Roberta Wallace and the staff of the Milwaukee County Zoo for their time and effort in collecting and sending samples. Tom Foose of the International Rhino Foundation and The Wilds for providing studbook information and current locations of black rhinoceroses, and Eric Lumis Shapiro of the Hospital of the University of Pennsylvania, Department of Anesthesiology, for reviewing this manuscript. Special thanks go to Lonnie McCaskill and Tortoise at White Oak Conservation Center for providing the inspiration to begin this work.
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