Glycosphingolipids from bovine milk and milk fat globule membranes: a comparative study. Adhesion to enterotoxigenic <i>Escherichia coli</i> strains

Sánchez-Juanes, Fernando; Alonso, Josefa María; Zancada, L.; Hueso, Pablo

doi:10.1515/bc.2009.003

Cited by 52 publications

(45 citation statements)

References 46 publications

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“…From this original bacterial suspension, several dilutions were made to find the minimum number of CFU required to agglutinate erythrocytes. Hemagglutination tests were carried out at 4 °C on sterile V-shaped 96-well plates (Nalge Nunc International, Roskilde, Denmark), as previously reported (Sánchez-Juanes et al, 2009b). We assayed individual and total PL (25 and 50 µg).…”

Section: Hemagglutination Assaysmentioning

confidence: 99%

Phospholipid classes and fatty acid composition of ewe’s and goat’s milk

et al. 2013

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RESUMEN Contenido de ácidos grasos de las diferentes clases de fosfolípidos de la leche de oveja y de cabraSe ha analizado el contenido, distribución de las especies individuales y la composición en ácidos grasos de los fosfolípidos (FL) de la leche de oveja y de cabra. Se ha estudiado también la unión de cepas enterotoxigénicas y uropatogénicas de Escherichia coli a estos compuestos y el efecto de los FL sobre la hemaglutinación provocada por estas bacterias mediante inmunodetección en placa y ensayos en placas multipocillo, respectivamente. La leche de oveja y de cabra contiene más FL que la de vaca, pero menos que la leche humana. El perfil de FL individuales es similar en la leche de oveja y de cabra e incluye esfingomielina, fosfatidilcolina, fosfatidiletanolamina, fosfatidilserina y fosfatidilinositol. En cuanto a la composición en ácidos grasos, los FL de ambos tipos de leche presentan un contenido elevado de ácidos grasos de cadena larga (más de 16 carbonos) y no saturados, siendo el más abundante C18:1. La leche de oveja contiene ácidos grasos más largos y menos saturados, mientras que los de la leche de cabra son más cortos y más saturados. No se ha encontrado adhesión de ninguna de las cepas bacterianas estudiadas a los FL individuales de la leche de oveja o de cabra .Tampoco se ha observado inhibición de la hemaglutinación producida por las bacterias por los FL. Estos compuestos son constituyentes fundamentales de la membrana de los glóbulos de grasa de la leche, pero a la vista de los resultados obtenidos, no parece que participen en la defensa del recién nacido frente a infecciones bacterianas. PALABRAS CLAVE: Ácidos grasos -Adhesión bacteriana -Fosfolípidos -Leche de cabra -Leche de oveja. SUMMARY Phospholipid classes and fatty acid composition of ewe's and goat's milksThe content, distribution of individual species, and the fatty acid composition of phospholipids (PL) from ewe's and goat's milk were analyzed. The binding of enterotoxigenic and uropathogenic Escherichia coli strains to PL and the inhibition of bacterial hemagglutination by PL were addressed using high performance thin-layer chromatography-overlay assays and microtiter plates, respectively. Ovine and caprine milk contained more PL than bovine milk but less than human milk. The profile of individual PL was similar, including sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol in both ovine and caprine milk. Regarding the fatty acid composition, a high content of long-chain fatty acids (more than C16) and unsaturated fatty acids, with C18:1 as the most abundant was found in ovine and caprine milk PL. Ovine milk has longer and less saturated fatty acids while caprine milk has shorter and more saturated ones. Neither the adhesion of any bacterial strains assayed to the individual PL from ovine or caprine milk nor the inhibition of bacterial hemagglutination by PL were observed. These are important constituents of the milk fat globule membrane, but it seems that they do not play a ro...

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Section: Hemagglutination Assaysmentioning

confidence: 99%

Phospholipid classes and fatty acid composition of ewe’s and goat’s milk

et al. 2013

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“…However, the neutral GSL fatty acid content seemed to be quite different. Whole milk had fewer very long-chain fatty acids (18.1 versus 46.4% in MFGMs) and more mediumchain and unsaturated C18:1 and C18:2 fatty acids [65].…”

Section: Gc-msmentioning

confidence: 95%

“…Starting from these observations, the GSL content of MFGMs and of the raw milk of Holstein-Friesian cows was investigated [65]. Although the GSL fractions isolated from milk were evaluated by TLC and densitometric analysis, the fatty acid contents of GSL were studied by GC-MS, after methanolysis with hydrochloric acid (20 h at 801C).…”

Section: Gc-msmentioning

confidence: 99%

High‐performance separation techniques hyphenated to mass spectrometry for ganglioside analysis

et al. 2011

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Gangliosides, sialic-acid-containing glycosphingolipids are involved in numerous biological processes and play essential roles in severe pathologies, with predilection in those of the central nervous system. Formerly, ganglioside composition and quantity were assessed exclusively by thin-layer chromatographic (TLC), immunochemical, and immunohistochemical methods, which have limited effectiveness being unable to detect minor components in mixtures of high heterogeneity. Increased awareness of the biological importance of gangliosides stimulated the development of analytical methods that are better amenable to complex ganglioside mixtures. More recently, MS in online conjunction with high-performance separation techniques brought a significant progress to the field. This review highlights the state-of-the-art development and application of separation methods online coupled to MS for ganglioside analysis. Most original and successful protocols based on GC-MS, LC-MS, and CE-MS are presented here together with the special instrumental and sample preparation requirements to be met for effective ganglioside separation, detection, and structural identification. Finally, the advantages and downsides of each methodology as well as the perspectives for simplification, standardization, and upgrading are assessed.

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“…As regards dairy products, due to their complex composition, high fat content and gangliosides located in the membrane surrounding fat globules, more complex lipid extraction is required (acetone/chloroform:methanol in different proportions) [45,62,63].…”

Section: Extraction Isolation and Purification Of Gangliosidesmentioning

confidence: 99%

“…Total gangliosides could be quantified in isolated and purified fraction determining the sialic acids content (see Section 2.2)-the results being expressed as lipid bound sialic acid (LBSA) [45,63].…”

Section: Determination Of Gangliosidesmentioning

confidence: 99%