Calf intestinal mucin: Isolation, partial characterization, and measurement in heal digesta with an enzyme-linked immunosorbent assay

Abstract

The aim of this study was to develop a specific ELISA for calf intestinal mucin to quantify its contribution to ileal endogenous losses. Mucin was isolated from intestinal scrapings by cesium chloride density gradient ultracentrifugation. The isolated mucin had a high concentration of glutamic and aspartic acids, threonine, and serine (13.2, 11.2, 9.6, and 9.2 mol % of total amino acids assayed, respectively). The carbohydrates present were (mol % of total hexoses):galactose 42.1, N-acetylglucosamine 24.1, N-acetylgalactosamine 23.6, fucose 4.7, mannose 3.1, and sialic acids 2.4. Amino acids and carbohydrates represented 52.6 and 47.4% of the mucin by weight, respectively. A rabbit hyperimmune plasma was raised against purified mucin and used to set up an ELISA. The linear range of this assay was 20 to 640 ng/ml. The plasma cross-reacted with calf abomasal and colonic mucin. It showed no cross-reactivity with nonmucin components and no reactivity with intestinal mucin from other animal species except for the rat. Mucin accounted for approximately 3.5% of the dry matter output at the ileum of calves fed a substitute milk based on skim milk powder. This represented a flow of 3.4 g of mucin/kg of dry matter intake. Mucin flow increased when dietary protein was provided by cow's colostrum. Finally, the developed assay is a suitable tool to investigate the impact of dietary factors on the flow of mucin along the gut of preruminant calves.