Survival of Human Milk Oligosaccharides in the Intestine of Infants

Chaturvedi, Prasoon; Warren, Christopher D.; Buescher, Christine R.; Pickering, Larry K.; Newburg, David S.

doi:10.1007/978-1-4615-1371-1_39

Cited by 107 publications

(110 citation statements)

References 8 publications

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“…Some analytical studies have indicated that there may be as much as 20 g/L of oligosaccharides in breast milk which would make them the third most concentrated component after lactose and fat [1,2]. However, unlike lactose and fat, these molecules do not appear to provide energy to infants as they resist the action of human digestive enzymes [3,4]. Most human milk oligosaccharides (HMO) are elongation products of lactose, and are synthesized from glucose, galactose, glucosamine, fucose and sialic acid [2].…”

Section: Introductionmentioning

confidence: 99%

Isolation of Milk Oligosaccharides using Solid-Phase Extraction

Ward¹

2009

TOGLYJ

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Abstract:A method was developed to isolate mono-and disaccharide-free oligosaccharides from human and bovine milk using a combination of enzymatic digestion of lactose and solid-phase extraction. In the initial trial, 2.5 g of oligosaccharides were isolated from one liter of human milk. In subsequent trials this was increased to over 5 g of oligosaccharides per liter. Compared to filtration-based extraction methods, this procedure allows for further isolation of oligosaccharide fractions via modulation of the column rinsing solvent. Neutral monosaccharide composition of the oligosaccharide polymers was investigated using gas chromatographic analysis of the monosaccharides as alditol acetate derivatives. Results indicate oligosaccharides are approximately made up of 24% fucose, 41% galactose, 22% glucose and 13% glucosamine. Isolated bovine and human milk oligosaccharides were compared to lactose as fermentation substrates for Bifidobacterium longum biovar infantis. Lactose fermentation yielded the greatest production of biomass followed by bovine and human milk oligosaccharides.

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Section: Introductionmentioning

confidence: 99%

Isolation of Milk Oligosaccharides using Solid-Phase Extraction

Ward¹

2009

TOGLYJ

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“…HMO are the third most abundant component of human milk (9), and at least 130 different masses have been identified (14). In vitro, HMO are resistant to catabolism by host hydrolases (3,4), and based on the mass balance between consumption and excretion, Chaturvedi et al (1) calculated that 97% of HMO pass through infants undigested, while Coppa et al (2) estimated that 40 to 50% of HMO pass through infants undigested.…”

mentioning

confidence: 99%

In Vitro Fermentation of Breast Milk Oligosaccharides by Bifidobacterium infantis and Lactobacillus gasseri

Ward¹,

Niñonuevo²,

Mills

et al. 2006

Appl Environ Microbiol

266

229

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It has been proposed that human milk oligosaccharides (HMO) function as a prebiotic for bifidobacteria, yet this activity has not been adequately investigated. In this study, Bifidobacterium infantis was shown to ferment purified HMO as a sole carbon source, while another gut commensal, Lactobacillus gasseri, did not ferment HMO. Our results support the hypothesis that HMO selectively amplify bacterial populations in the infant intestine.Human milk is unique because of the high concentration and diversity of human milk oligosaccharides (HMO). HMO are the third most abundant component of human milk (9), and at least 130 different masses have been identified (14). In vitro, HMO are resistant to catabolism by host hydrolases (3, 4), and based on the mass balance between consumption and excretion, Chaturvedi et al.(1) calculated that 97% of HMO pass through infants undigested, while Coppa et al. (2) estimated that 40 to 50% of HMO pass through infants undigested.A prebiotic function was attributed to HMO based on studies done in the 1950s with Bifidobacterium bifidus subsp. pennsylvannicus (6, 7), yet the measurements were based on growth enhancement, not fermentability, as the media contained lactose. Direct fermentation of HMO by Bifidobacterium spp. and/or Lactobacillus spp. has not been demonstrated yet, and there are many questions about the metabolic fate of these molecules. Do subpopulations mediate specific functions, such as pathogen binding or prebiotic activity? Are specific bacterial species required for catabolism in the gut? The goals of this investigation were to determine the fermentability of HMO by two representative species of breast-fed infant microbiota, Bifidobacterium infantis and Lactobacillus gasseri (8,10) and to characterize the changes in the HMO after bacterial growth. Determination of the biological basis underlying the HMO abundance in human milk should be of general interest in human nutrition (15).Pooled milk was provided by the Mother's Milk Bank of San Jose, CA, and the Mother's Milk Bank of Austin, TX. Oligosaccharides were extracted as described by Gnoth et al. (5), with modifications. One liter of milk was centrifuged at 5,000 ϫ g for 30 min at 4°C, and the fat was removed. Ethanol (2 liters) was added, and the solution was incubated overnight at 4°C. The precipitate was removed by centrifugation at 5,000 ϫ g for 30 min at 4°C, and the solvent was removed by rotary evaporation. The concentration of the solution was adjusted to 0.05 M with phosphate buffer (pH 6.8), 3,000 U ␤-galactosidase (Kluyveromyces fragilis) was added, and the solution was incubated for 1 h at 37°C. Then the solution was extracted with 4 volumes of chloroform-methanol (2:1, vol/vol), and the aqueous layer was collected.As described by Redmond and Packer (12), monosaccharides and disaccharides were removed by selective adsorption of HMO, using solid-phase extraction with graphitized nonporous graphitized carbon cartridges (Supelco Inc., Bellefonte, PA). The oligosaccharides that were retained were eluted wit...

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“…Each single oligosaccharide varies dynamically during the different phases of lactation (9). From the quantitative point of view, oligosaccharides, all together, represent the third component of human milk, besides lactose and lipids (10,11).Breast-fed infants ingest daily several grams of oligosaccharides and these substances have been found in feces in large quantities (12)(13)(14). It follows that most oligosaccharides contained in human milk are not digested by intestinal enzymes, and so are present in large quantity both in the small and large intestine of neonates (15,16).…”

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confidence: 99%

Human Milk Oligosaccharides Inhibit the Adhesion to Caco-2 Cells of Diarrheal Pathogens: Escherichia coli, Vibrio cholerae, and Salmonella fyris

et al. 2006

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Breast-fed children, compared with the bottle-fed ones, have a lower incidence of acute gastroenteritis due to the presence of several antiinfective factors in human milk. The aim of this work is to study the ability of human milk oligosaccharides to prevent infections related to some common pathogenic bacteria. Oligosaccharides of human milk were fractionated by gel-filtration and characterized by thin-layer chromatography and highperformance anion exchange chromatography. Fractions obtained contained, respectively, 1) acidic oligosaccharides, 2) neutral highmolecular-weight oligosaccharides, and 3) neutral low-molecularweight oligosaccharides. Experiments were carried out to study the ability of oligosaccharides in inhibiting the adhesion of three intestinal microorganisms (enteropathogenic Escherichia coli serotype O119, Vibrio cholerae, and Salmonella fyris) to differentiated Caco-2 cells. The study showed that the acidic fraction had an antiadhesive effect on the all three pathogenic strains studied (with different degrees of inhibition). The neutral high-molecular-weight fraction significantly inhibited the adhesion of E. coli O119 and V. cholerae, but not that of S. fyris; the neutral low-molecular-weight fraction was effective toward E. coli O119 and S. fyris but not V. cholerae. Our results demonstrate that human milk oligosaccharides inhibit the adhesion to epithelial cells not only of common pathogens like E. coli but also for the first time of other aggressive bacteria as V. cholerae and S. fyris. Consequently, oligosaccharides are one of the important defensive factors contained in human milk against acute diarrheal infections of breast-fed infants. (Pediatr Res 59: 377-382, 2006) I nfectious diarrheal diseases constitute a leading cause of infant morbidity not only in developing countries but also in developed areas (1). In fact, in the United States, acute diarrhea represents an important cause of childhood morbidity, especially during the first years of life, determining an economical loss that has been estimated in the order of several millions of dollars (2).In infants, infections of the gastrointestinal tract are caused by a wide variety of enteropathogens, including bacteria, viruses, and parasites. There is now strong evidence supporting a relationship between breast-feeding and a lower incidence in diarrhea. Breast-feeding offers protection with different mechanisms against diarrhea due to several antiinfective substances present in human milk, such as secretory antibodies, lactoferrin, lysozyme, etc. (3-5).In the last 15 y, evidence has also emerged on the protective role of another group of substances, oligosaccharides (6 -8). They are synthesized in a large number by specific glycosyltransferases present in the mammary gland through the sequential addition to lactose of fucose, galactose, Nacetylglucosamine, and sialic acid. Each single oligosaccharide varies dynamically during the different phases of lactation (9). From the quantitative point of view, oligosaccharides, all tog...

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Survival of Human Milk Oligosaccharides in the Intestine of Infants

Cited by 107 publications

References 8 publications

Isolation of Milk Oligosaccharides using Solid-Phase Extraction

Isolation of Milk Oligosaccharides using Solid-Phase Extraction

In Vitro Fermentation of Breast Milk Oligosaccharides by Bifidobacterium infantis and Lactobacillus gasseri

Human Milk Oligosaccharides Inhibit the Adhesion to Caco-2 Cells of Diarrheal Pathogens: Escherichia coli, Vibrio cholerae, and Salmonella fyris

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