Determination of serum bone-specific alkaline phosphatase isoenzyme activity in captive Asian elephants (<i>Elephas maximus</i>) using an agarose gel electrophoresis method

Abstract

The bone-specific alkaline phosphatase (ALP) isoenzyme activity was measured in 51 serum samples from four captive Asian elephants ( Elephas maximus ) using a conventional method with wheat germ lectin precipitation and a commercial agarose gel electrophoresis (AGE) kit; the isoenzymes were designated as bone-specific ALP (BAP) and ALP isoenzyme 3 (ALP3), respectively. This study examined the suitability of the AGE kit for analyzing blood biochemistry in Asian elephants. The serum ALP3 and BAP activities were strongly positively correlated and met the evaluation criteria for agreement using Bland-Altman analysis. The results indicate that the AGE kit can be used to examine the blood biochemistry in Asian elephants instead of the conventional method.

The serum activities of alkaline phosphatase isoenzymes measured using two approved methods in zoo-managed Asian elephants (<i>Elephas maximus</i>)

“…We previously reported that measurements of ALP and its bone-derived isoenzyme (ALP3) in blood aided assessments of bone growth and health in elephant calves [13].…”

“…Serum total ALP (t-ALP) levels was measured using a blood chemistry analyzer (DRI-CHEM 4000sV; Fujifilm Medical, Tokyo, Japan) employing a DRI-CHEM slide v-ALP-P (JSCC method) (Fujifilm Medical) or an ALP-P slide (IFCC method) (Fujifilm Medical). ALP2 and ALP3 levels were determined using agarose gel electrophoresis (AGE) [13] employing a QuickGel ALP agarose gel kit (J713; Helena Laboratories Japan, Saitama, Japan), QuickGel ALP (bone type) reagent (JSCC) (J871; Helena Laboratories), QuickEP ALP-IF reagent (IFCC) (J890; Helena Laboratories), and a fully automatic electrophoresis system (Epalyzer-2; Helena Laboratories). Briefly, each serum sample (30 µL) was incubated with 4 µL of a 1.5 U/mL protease solution and 2 µL of a separator solution (containing neuraminidase) at room temperature for 30 min.…”

“…The circulating ALP of healthy individuals features several isoenzymes (ALP2, ALP3, and ALP5; the latter is intestine-derived ALP). The predominant isoenzyme differs among species; ALP2 is the major isoenzyme in dogs [6] and cats [8] but ALP3 in cattle [4] and Asian elephants [13]. We earlier found that ALP5 was undetectable in elephant sera [13] but was evident in bovine sera [4].…”

Anthropometric and blood data on a hand-reared captive Asian elephant (<i>Elephas maximus</i>) calf: A retrospective case report

“…The serum biochemistry included the alanine transaminase (ALT), aspartate aminotransferase (AST), calcium (Ca), inorganic phosphorus (iP), glucose (Glu), total cholesterol (t-Cho), blood urea nitrogen (BUN), total alkaline phosphatase (t-ALP), and total protein (TP) levels, which were determined using a blood chemistry analyzer (DRI-CHEM 4000sV; FUJIFILM Medical, Tokyo, Japan). Additionally, frozen (−20°C) serum samples of calves A, 3, and 4 were used to measure the alkaline phosphatase isoenzyme 3 (ALP3) [20] and tartrate-resistant acid phosphatase 5b (TRAP 5b) activities [19] as bone metabolism markers. Protein electrophoretic analysis was done using a QuickGel SP (J711, Helena Laboratory Japan, Saitama, Japan).…”

“…Circulating bone metabolism markers in Asian elephants are useful indices of bone metabolism [2,19,20,22]. ALP3 is a t-ALP isoenzyme that reflects osteoblast function [5,17].…”