Quantitative determination of equine alkaline phosphatase isoenzymes in foal and adult serum.
Abstract: Automated and semiautomated assays were developed and validated for the determination of equine alkaline phosphatase isoenzymes including intestinal (IALP), bone (BALP), and liver (LALP). The addition of levamisole selectively inhibited more than 97% of LALP while inhibiting only 55% of IALP. Because these percentages were highly reproducible in an automated system, the IALP activity could be calculated in a sample. Bone alkaline phosphatase isoenzyme was selectively precipitated by adding an equal volume of wheat germ agglutinin (5 mg/mL), incubating for 30 minutes at 37 degrees C, and centrifugating. The LALP activity was determined from the supernatant fluid and BALP activity was calculated by subtraction from total ALP activity. The within-run coefficient of variation for determination of BALP activity was 4.7%. These assays were used to identify and quantify the isoenzymes present in pony foal sera through the first 21 days of life, in horse foal sera before colostrum ingestion and during the first 21 days of life, and in adult horse and pony sera. Intestinal ALP activity was not found in sera of any of the foals or adult ponies or horses. A majority of serum ALP activity of newborn foals is of bone origin (80 to 92%) which decreases markedly over the first 21 days. In adults, only 17.9% (51.2 +/- 18.1 U/L) of serum ALP is derived from bone. The absolute LALP activity in foal serum is similar to that in adults.
Publication Date: 1993-01-01 PubMed ID: 8455179DOI: 10.1111/j.1939-1676.1993.tb03164.xGoogle Scholar: Lookup
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- Journal Article
Summary
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The research article presents the development and validation of methods to determine the specific types and amounts of alkaline phosphatase enzymes in horses, particularly in young and adult horses. The study also explores how these enzyme levels change as the animal matures.
Development and Validation of Assays
- Assays were designed to identify and quantify three types of enzymes, namely intestinal alkaline phosphatase (IALP), bone alkaline phosphatase (BALP), and liver alkaline phosphatase (LALP).
- These assays were made possible by using levamisole which selectively inhibits LALP and IALP to varying degrees which allows for the measurement of each enzyme’s activity.
- For BALP, a process utilizing wheat germ agglutinin was developed. This process involves precipitation, incubation, and centrifugation. The remaining LALP activity was then determined from the supernatant fluid, and the BALP activity was calculated by subtraction from the total ALP activity.
Application of Assays
- The assays were used to identify and quantify these enzymes in both young (foal) and adult horse serum, considering the first 21 days of life for the foals, and before and after the ingestion of colostrum by horse foals.
- No intestinal ALP activity was found in the serum of any of the foals, or adult ponies or horses.
Findings on ALP Isoenzymes Variation
- In newborn foals, the majority of their serum ALP activity (80 to 92%) came from bone. This marker of bone growth decreased significantly over the first 21 days, implying rapid bone development immediately after birth.
- In adult horses, the bone-derived ALP only comprises about 18% of serum ALP showing that bone growth substantially slows down in adulthood.
- The research also discovered that the absolute LALP (liver enzyme) activity in foal serum is similar to that in adults, which means the liver is functional at an early age.
Cite This Article
APA
Hank AM, Hoffmann WE, Sanecki RK, Schaeffer DJ, Dorner JL.
(1993).
Quantitative determination of equine alkaline phosphatase isoenzymes in foal and adult serum.
J Vet Intern Med, 7(1), 20-24.
https://doi.org/10.1111/j.1939-1676.1993.tb03164.x Publication
Researcher Affiliations
- College of Veterinary Medicine, University of Illinois, Urbana.
MeSH Terms
- Alkaline Phosphatase / blood
- Animals
- Animals, Newborn / blood
- Bone and Bones / enzymology
- Horses / blood
- Intestinal Mucosa / enzymology
- Isoenzymes / blood
- Liver / enzymology
- Reproducibility of Results
- Sensitivity and Specificity
Citations
This article has been cited 4 times.- Drozdzewska K, Gehlen H. Markers for internal neoplasia in the horse. Vet Med Sci 2023 Jan;9(1):132-143.
- Jia Y, Son K, Burris WR, Bridges PJ, Matthews JC. Forms of selenium in vitamin-mineral mixes differentially affect serum alkaline phosphatase activity, and serum albumin and blood urea nitrogen concentrations, of steers grazing endophyte-infected tall fescue. J Anim Sci 2019 May 30;97(6):2569-2582.
- De Palo P, Maggiolino A, Albenzio M, Casalino E, Neglia G, Centoducati G, Tateo A. Survey of biochemical and oxidative profile in donkey foals suckled with one natural and one semi-artificial technique. PLoS One 2018;13(6):e0198774.
- Carrillo-Muro O, Hernández-Briano P, Correa-Aguado PI, Rivera-Villegas A, Sánchez-Barbosa OY, Lazalde-Cruz R, Barreras A, Plascencia A, Rodríguez-Cordero D. Enzymic Activity, Metabolites, and Hematological Responses Changes of Clinical Healthy High-Risk Beef Calves During Their First 56-Days from Arrival. Animals (Basel) 2025 Jan 8;15(2).
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