Samuel Martín-Sosa

Bovine milk gangliosides: Changes in ceramide moiety with stage of lactation

The stage of lactation is one of the most important factors that influence milk composition. Changes in fatty acids from triacylglycerols and phospholipids have already been reported. In this study, we looked for a lactational change in the ganglioside lipid moeity since ganglioside contents and patterns vary strongly with stage of lactation. Individual gangliosides from four stages were isolated, methanolyzed to cleave the bonds between individual constituents, and derivatized for gas-liquid chromatography and gas chromatography/mass spectrometry analyses. Ceramide components, both fatty acids (as methyl esters derivatives) and long-chain bases, were identified and quantified. The results pointed to a marked change in ceramide from colostrum to milk that was characterized by a dramatic decrease in saturated and the longest-chain fatty acids as well as an increase in 18∶1 and 18∶2. The major long-chain base along lactation was a recently described structure, 3-ethoxy-15∶0 sphinganine. Other new long-chain base structures appeared in these gangliosides. All these changes suggest differences in the fluidity of the fat globule membrane, reflecting physiological variations in cows with respect to milk production.

Distribution of sialic acids in the milk of spanish mothers of full term infants during lactation.

Objectives: The protective effect of human milk against infection is well known. Several non-immunologic components, including complex carbohydrates, have been described. The present study was undertaken to determine the sialic acid distribution in different milk fractions (complex carbohydrates). Methods: Milk samples from 12 Spanish women at three different lactational stages (colostrum, transitional milk and mature milk) were analyzed. Total and glycoprotein-bound, oligosaccharide-bound, casein-bound, and lipid-bound sialic acids were determined. Results: Sialic acids from human milk are mainly bound to oligosaccharides and only a small amount is present bound to glycoproteins or in the free form. All the fractions analyzed showed a similar trend: sialic acids decrease rapidly along lactation. Casein-bound sialic acid does not follow this trend. We detected the presence of an O-acetylated species of N-acetylneuraminic acid. Conclusions: In human milk from Spanish women we observed slightly different values than those previously reported. This could be a result of population differences but nutritional or methodological aspects can not be discarded.

Binding of milk oligosaccharides by several enterotoxigenic Escherichia coli strains isolated from calves

Milk oligosaccharides have been proposed to play an important role in newborn defense, blocking bacterial adhesion to the intestinal mucosa and preventing infections. Some studies have been performed on human milk oligosaccharides. Here we checked whether bovine milk oligosaccharides would achieve the same protective action against the most common calf enteric pathogens. Seven enterotoxigenic Escherichia coli strains, isolated from diarrheic calves, were selected. All strains managed to agglutinate horse erythrocytes, and we therefore used the inhibition of hemagglutination in the presence of oligosaccharides as an indicator of the union between oligosaccharide and bacterial adhesins. Oligosaccharides from different stages of bovine lactation and standard oligosaccharides were assayed. Midlactation milk, in particular that corresponding to the transition period, proved to be the most efficient at inhibiting hemagglutination. The standard oligosaccharides used pointed to the preference of several strains (K99-, F41-, and F17-fimbriated) for α2,6-linked sialic acid. By contrast, B23 fimbriae exhibited higher affinity for α2,3-sialylated isomers and B64 seemed to require N-acetylglucosamine for binding.Our results suggest a general trend for milk oligosaccharides. Probably they participate in the protection of newborn mammals from pathogens.

Enterotoxigenic Escherichia coli strains bind bovine milk gangliosides in a ceramide-dependent process

Diarrhea caused by enterotoxigenic Escherichia coli (ETEC) is the main infectious disease of newborn calves. The first step of infection involves bacterial attachment to the intestinal mucosa. This adhesion is mediated by fimbriae that recognize some glycoconjugates on the host cell surface, in particular, several gangliosides. Because milk also contains gangliosides, these have been suggested to serve as ligands for bacterial fimbriae and thus prevent the bacterial attachment to mucosa. The most relevant ETEC strains in calves, including those with K99 and F41 fimbriae, were assayed to determine whether they are able to bind gangliosides isolated from several stages of bovine lactation. Both GM3 and GD3, the main gangliosides of milk, were recognized by ETEC strains, although the different fimbriae showed diverse levels of affinity. Unexpectedly, the adhesion to colostral gangliosides was considerably weaker than that to gangliosides from the other stages of lactation. Because the carbohydrate moiety did not change and because differences in the percentages of unsaturated FA and sphingosine between colostrum and other stages were observed, we conclude that the differences in adhesion could be due to a different composition of the ganglioside caramide.