Protection against indomethacin-induced loss of intestinal epithelial barrier function by a quercetin oxidation metabolite present in onion peel: In vitro and in vivo studies.

Oxidative stress is directly implicated in the loss of intestinal epithelial barrier function (IEBF) induced by non-steroidal anti-inflammatory drugs (NSAIDs). Previous studies by our research team demonstrated that 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxy-3(2H)-benzofuranone (BZF), a quercetin oxidation metabolite that naturally occurs in onion peels, exhibits an antioxidant potency notably higher than quercetin. Thus, we assessed the potential of BZF and a BZF-rich onion peel aqueous extract (OAE) to protect against the loss of IEBF in Caco-2 cell monolayers and in rats exposed to indomethacin. In vitro, pure BZF and OAE standardized in BZF (100 nM), protected against the drop in transepithelial electrical resistance by 70 - 73%. Likewise, it prevented the increase in fluorescein-isothiocyanate labelled dextran (FITC-dextran) paracellular transport by 74% and oxidative stress by 84 - 86%. In vivo, BZF, given orally at a dose 80 µg/Kg bw as OAE, totally abolished a 30-fold increase in FITC-dextran serum concentration induced by indomethacin. This effect was dose-dependent and largely conserved (85%) when OAE was given 180-min prior to indomethacin. The IEBF-protective effect of OAE was accompanied by a full prevention of the NF-ĸB activation, and the increases in interleukine-8 secretion and myeloperoxidase activity induced by indomethacin. The protection was also associated with a 21-fold increase in Nrf2, and a 7-fold and 9-fold increase in heme oxygenase-1 and NAD(P)H-quinone oxidoreductase 1, respectively. The IEBF-protecting effect of OAE involves, most likely, its dual capacity to activate Nrf2 while inhibiting NF-ĸB activation. The extremely low doses of BZF needed to promote such actions warrants extending its IEBF-protective effects to other NSAIDs.

[Advances in the knowledge about human milk proteins]. / Avances en el conocimiento de las proteínas de la leche materna.

The mammary gland and maternal milk are the product of millions of years of evolution that resul ted in an optimal composition that sustains the growth and development of newborns and infants. Maternal milk supports the growth, adaptation and survival of this immature organism. Recent studies have detected 1606 different proteins in human milk, most of them synthesized in the acini of the glandular tissue while others originate from distant organs such as the lymphoid tissue and the digestive tract. Maternal milk enzymes modify its proteins and liberate peptides with antimicrobial, antihypertensive or stimulatory activities. This proteolytic activity occurs at specific sites in peptide chains. To prevent the extemporaneous activation of these proteolytic enzymes, that would result in inflammatory processes, maternal milk also contains inhibitory peptides that together with the stimulatory peptides conform a complex regulatory system. Some enzymes in maternal milk main tain their activity in the gastrointestinal tract of infants and compensate for the decreased activity of digestive tract enzymes in newborns. Thus, the milk enterokynase stimulates the release of pancreatic proteases as it induces the liberation of cholecystokynin/pancreozymin. The bile salt-activated lipase of human milk is activated in the duodenum by the infants' bile salts and partially compensates for the low levels of pancreatic lipase in newborns. These milk enzymes probably contribute to the nutrition of premature infants as they increase the availability of amino acids and peptides in their upper gastrointestinal tract; furthermore, as their intestinal epithelium is more permeable to peptides and partially digested protein this may help induce immune tolerance. The most relevant issues in the physiology and composition of the maternal milk are presented in this review.

Avances en el conocimiento de las proteínas de la leche materna / Advances in the knowledge about human milk proteins

La glándula mamaria y la leche materna son el resultado de millones de años de una evolución que llevó a una composición óptima para el crecimiento y desarrollo de recién nacidos y lactantes; la leche materna favorece el crecimiento, la adaptación y la supervivencia de su organismo y de sus órganos inmaduros. Análisis recientes han demostrado en ella la presencia de 1606 proteínas que en su mayoría son sintetizadas en los acinos de la glándula mamaria aunque otras proteínas y péptidos provienen de órganos como el sistema linfático y el aparato digestivo. La composición de la leche materna incluye enzimas que modifican sus proteínas y originan péptidos antimicrobianos, antihipertensivos y estimuladores del metabolismo. Esta actividad proteolítica actúa en sitios específicos de las cadenas peptídicas de la proteína de la leche. La activación extemporánea de estos enzimas en los acinos es regulada por péptidos inhibidores y activadores que previenen procesos inflamatorios. Algunos enzimas de la leche actúan en el tubo digestivo de recién nacidos y lactantes y complemen tan la menor concentración y actividad de sus propios enzimas digestivos. Así, la enteroquinasa de la leche estimula la liberación de enzimas pancreáticos (mediada por el estímulo de la colecistoquinina-pancreozimina); la lipasa activada por las sales biliares complementa la baja producción de lipasa pancreática. Estas actividades probablemente facilitan la nutrición de los prematuros, cuyo tubo di gestivo es más permeable a las proteínas parcialmente hidrolizadas y cuyas actividades enzimáticas y factores defensivos locales no han alcanzado su plena madurez. Esto también puede estimular en ellos la tolerancia inmunológica. En este artículo se presentan los aspectos fisiológicos relevantes de la leche materna, y los avances en el conocimiento de su composición, para el cabal conocimiento del pediatra de esta importante materia.

The mammary gland and maternal milk are the product of millions of years of evolution that resul ted in an optimal composition that sustains the growth and development of newborns and infants. Maternal milk supports the growth, adaptation and survival of this immature organism. Recent studies have detected 1606 different proteins in human milk, most of them synthesized in the acini of the glandular tissue while others originate from distant organs such as the lymphoid tissue and the digestive tract. Maternal milk enzymes modify its proteins and liberate peptides with antimicrobial, antihypertensive or stimulatory activities. This proteolytic activity occurs at specific sites in peptide chains. To prevent the extemporaneous activation of these proteolytic enzymes, that would result in inflammatory processes, maternal milk also contains inhibitory peptides that together with the stimulatory peptides conform a complex regulatory system. Some enzymes in maternal milk main tain their activity in the gastrointestinal tract of infants and compensate for the decreased activity of digestive tract enzymes in newborns. Thus, the milk enterokynase stimulates the release of pancreatic proteases as it induces the liberation of cholecystokynin/pancreozymin. The bile salt-activated lipase of human milk is activated in the duodenum by the infants' bile salts and partially compensates for the low levels of pancreatic lipase in newborns. These milk enzymes probably contribute to the nutrition of premature infants as they increase the availability of amino acids and peptides in their upper gastrointestinal tract; furthermore, as their intestinal epithelium is more permeable to peptides and partially digested protein this may help induce immune tolerance. The most relevant issues in the physiology and composition of the maternal milk are presented in this review.

[Probiotics: innocuousness, prevention and risks]. / Inocuidad, prevención y riesgos de los probióticos.

Probiotics have been defined as live microorganisms which, when ingested in adequate numbers, confer health benefits to the host. They are currently consumed without any age restrictions and adverse effects such as sepsis, a marker of the risk of invasion of the bloodstream, are extremely infrequent. However, some health professionals express doubts about probiotics being truly innocuous. This review discusses the incidence of sepsis secondary to probiotics use, mainly lactobacilli and bifidobacteria, evaluated through molecular biology or classic culture techniques, showing that sepsis in large numbers of individuals along decennia is extremely low, of the order of 0,02% en some centers or as low as 1 case/million population in France. These data are important considering the use different species and strains of these microorganisms. Few studies which have reported other adverse effects but many of these have problems with their design that cast doubt about the validity of their results. On the contrary, it has been shown that probiotic microorganisms exert positive stimulatory effects on innate and acquired immunity, with decrease of the manifestations of atopy and eczema. These positive effects are further evidenced by the beneficial effects of many species of probiotics in preventing necrotizing enterocolitis in patients as functionally labile as premature-born babies.

Inocuidad, prevención y riesgos de los probióticos / Probiotics: innocuousness, prevention and risks

Los probióticos son microorganismos vivos que cuando son administrados en cantidades adecuadas confieren beneficios a la salud del huésped. Su empleo es muy amplio en todas las edades, y los episodios adversos asociados con ellos, incluyendo casos de sepsis, que hagan suponer un riesgo real y manifiesto de invasión del torrente sanguíneo o algún tipo de contagio son muy infrecuentes. Sin embargo, algunos profesionales de la salud manifiestan dudas respecto de su inocuidad. Esta revisión revela que la incidencia de sepsis por probióticos, principalmente lactobacilos y bifidobacterias, evaluada en casuísticas numerosas a lo largo de decenios usando métodos de biología molecular o cultivos, es muy baja: del orden de 0,021% en algunas estadísticas y podría llegar hasta 1 caso/1.000.000 de habitantes. Estos datos son importantes teniendo en cuenta el empleo masivo de diversas especies y cepas, sin restricciones de ninguna especie y en todas las edades. Algunos estudios que comunican otros efectos adversos tienen problemas de diseño que ponen en duda su validez. Por el contrario, se ha demostrado que los probióticos ejercen efectos positivos estimulando la inmunidad innata y adquirida, en el tratamiento de la atopia y eccema y en la prevención de la enterocolitis necrosante en sujetos tan vulnerables como los prematuros.

Probiotics have been defined as live microorganisms which, when ingested in adequate numbers, confer health benefits to the host. They are currently consumed without any age restrictions and adverse effects such as sepsis, a marker of the risk of invasion of the bloodstream, are extremely infrequent. However, some health professionals express doubts about probiotics being truly innocuous. This review discusses the incidence of sepsis secondary to probiotics use, mainly lactobacilli and bifidobacteria, evaluated through molecular biology or classic culture techniques, showing that sepsis in large numbers of individuals along decennia is extremely low, of the order of 0,02% en some centers or as low as 1 case/million population in France. These data are important considering the use different species and strains of these microorganisms. Few studies which have reported other adverse effects but many of these have problems with their design that cast doubt about the validity of their results. On the contrary, it has been shown that probiotic microorganisms exert positive stimulatory effects on innate and acquired immunity, with decrease of the manifestations of atopy and eczema. These positive effects are further evidenced by the beneficial effects of many species of probiotics in preventing necrotizing enterocolitis in patients as functionally labile as premature-born babies.

Probiotic screening and safety evaluation of Lactobacillus strains from plants, artisanal goat cheese, human stools, and breast milk.

The aim of this study was to select autochthonous strains of Lactobacillus from stools of healthy infants and adults, human milk, artisanal goat cheese, and fruits and vegetables according to their probiotic properties and safety. From 421 strains of Lactobacillus isolated, 102 (24.2%) were shown to be tolerant to gastric pH and bile salts; they were used to determine their anti-Helicobacter pylori (agar diffusion assay), antioxidant (oxygen radical absorption capacity), and anti-inflammatory (inhibition of interleukin-8 release by tumor necrosis factor-α-stimulated HT-29 cells) activities as well as their ability to adhere to intestinal (Caco-2) and gastric (AGS) epithelial cells. Results obtained were compared with three commercial probiotic Lactobacillus rhamnosus GG, L. plantarum 299v, and L. johnsonii NCC533. The five strains most efficient according to these activities were subsequently identified by sequencing their 16S rRNA gene, their susceptibility to antibiotics was determined, and their safety evaluated in mice. One strain of L. plantarum was discarded due to the higher prevalence of liver bacterial translocation observed in the animals fed this strain. In conclusion, four autochthonous strains of L. rhamnosus were finally selected with probiotic properties and safety allowing their eventual use in human studies. These results contribute to increase the diversity of probiotic strains available for the development of nutraceuticals and functional foods.

Identification of Lactobacillus spp. in colostrum from Chilean mothers.

The biodiversity of Lactobacillus spp. in colostrum samples from 116 Chilean mothers was analyzed by PCR and 16S rDNA sequencing. Lactobacilli were isolated in 55.3% of the samples, with concentrations of 3.33 +/- 0.55 (log CFU/ml). The predominant species were L. plantarum (64%), L. fermentum (16%) and L. pentosus (9%). 28% of the isolated strains were resistant to gastric pH and bile salts, suggesting that they could be used as probiotics.

Effect of Helicobacter pylori infection on iron absorption in asymptomatic adults consuming wheat flour fortified with iron and zinc.

Helicobacter pylori infection could impair iron absorption from fortified products. The objective of the study was to determine the effect of H. pylori infection on iron absorption from asymptomatic adults consuming wheat flour fortified with iron and zinc. The (13)C urea breath test was used to assess H. pylori infection. Twenty-four H. pylori-positive and 26 H. pylori-negative volunteers completed the study. On day 1, the subjects were randomized to receive for breakfast bread fortified with either ferrous sulfate and zinc sulfate or ferrous fumarate and zinc oxide. Bread fortified with ferrous sulfate was labeled with (59)Fe as sulfate, and bread fortified with ferrous fumarate was labeled with (55)Fe as fumarate. On day 3, they received the other type of bread, with the respective tracers. On days 18-23, a proton pump inhibitor was administered to all subjects. On day 24, all subjects received bread fortified with ferrous fumarate and zinc oxide labeled with (55)Fe as fumarate. H. pylori prevalence was 77.6%. The geometric mean (±1 SD) of iron absorption was significantly higher for ferrous sulfate than fumarate (6.9 ± 2.9% vs. 0.5 ± 3.5%, p < 0.001). The H. pylori-negative subjects absorbed significantly more iron from bread fortified with either ferrous sulfate (10.5 ± 3.1% vs. 4.4 ± 2.2%, p < 0.001) or ferrous fumarate (0.6 ± 3.9% vs. 0.4 ± 3.1%, p < 0.001). Iron absorption was not significantly different between groups after administration of a proton pump inhibitor (0.3 ± 3.3% vs. 0.3 ± 2.7%, p = 0.11). H. pylori infection has a negative effect on iron absorption in asymptomatic adults consuming iron- and zinc-fortified wheat flour.

Trabajos de Investigación Increase of plasma fatty acids without changes in n-6/n-3-PUFA ratio in asymptomatic obese subjects / Aumento de las concentraciones plasmáticas de ácidos grasos sin cambio en el ratio de n-6/n-3 PUFA en sujetos obesos asintomáticos

Obesity is associated with a low grade inflammation which contributes to the development of insulin resistance and diabetes. The aim of this study was to assess the total saturated (SFAs), monounsaturated (MUFAs) and polyunsaturated fatty acids (PUFAs) in plasma from asymptomatic obese subjects and to determine the arachidonic/eicosapentanoic acid ratio [ARA/EPA] as a marker of inflammation, and its eventual association with ultrasensitive CRP. Fourteen obese (34.4 ± 11.1y.; BMI: 36.0 ± 4,5 kg/m2) and 12 normal-weight (30.6 ± 7.8y.; BMI: 23,6± 2,4 kg/m2) subjects were recruited and their plasma fatty acids were determined by gas chromatography. usCRP was higher in the obese subjects (p=0,01) and correlates with their body fat content. The percentages of SFAs, MUFAs, PUFAs were not affected in the obese subjects but their concentrations were increased, compared with the control group. However, no differences in the long chain PUFAs (DHA and EPA) concentrations or in the plasmatic ARA/EPA ratio were observed in these subjects. These observations do not support a relation between the ARA/EPA ratio and the presence of low grade inflammation evaluated by plasma usCRP in this group of asymptomatic obese subjects.

La obesidad está asociada con una inflamación de bajo grado que contribuye al desarrollo de la insulino-resistencia y de la diabetes. El objetivo de este estudio fue evaluar las concentraciones plasmáticas de ácidos grasos saturados (AGS), monoinsaturados (AGMI) y poliinsaturados (AGPI) en sujetos obesos asintomáticos y determinar el ratio ácido araquidónico/ácido eicosapentanoico [ARA/EPA] como un posible marcador de inflamación, con su eventual asociación con los niveles de proteína C reactiva ultrasensible (PCRus). Se reclutaron 14 sujetos obesos (34,4 ± 11.1 años; índice de masa corporal: 36,0 ± 4,5 kg/m2) y 12 normopeso (30,6 ± 7.8 años; índice de masa corporal: 23,6 ± 2,4 kg/m2); las concentraciones plasmáticas de ácidos grasos fueron determinados por cromatografía de gases. Los niveles de PCRus fueron más elevadas en los sujetos obesos (p=0,01) y correlacionaron con el contenido de grasa corporal. Los porcentajes relativos de AGS, AGMI, AGPI no se vieron afectados en los sujetos obesos, pero sus concentraciones plasmáticas se incrementaron en comparación con el grupo control. Sin embargo, no se observaron diferencias en las concentraciones de PUFAs de cadena larga (DHA, EPA y ARA) ni en el ratio ARA/EPA en los sujetos obesos. Estas observaciones no apoyan el uso del ratio ARA/EPA como un marcador de inflamación de bajo grado evaluada por PCRus en este grupo de sujetos obesos asintomáticos.

Apple peel polyphenols protect against gastrointestinal mucosa alterations induced by indomethacin in rats.

The stability of an apple peel polyphenol extract (APPE) with powerful antioxidant activity was evaluated under acidic conditions in vitro, and its protective effect against gastrointestinal damage was investigated in rats treated with indomethacin. The antioxidant activity of APPE remained stable at pH 2.0 for 4 h. In rats treated with indomethacin (40 mg/kg ig), the previous administration of APPE protected the gastric, intestinal, and colonic mucosa from oxidative stress by preventing increased malondialdehyde concentrations and decreasing the GSH/GSSG ratio. APPE also displayed anti-inflammatory effects by preventing neutrophil infiltration in the mucosa, as evidenced by the lower myeloperoxidase activity. These protective effects of APPE resulted in the prevention of macro- and microscopic damage and of barrier dysfunction along the gastrointestinal tract of the indomethacin-treated animals. This study supports the concept that apple peel polyphenols may be useful in the prevention and/or treatment of nonsteroidal anti-inflammatory drug-associated side effects.

Identification of Lactobacillus spp. in colostrum from Chilean mothers

The biodiversity of Lactobacillus spp. in colostrum samples from 116 Chilean mothers was analyzed by PCR and 16S rDNA sequencing. Lactobacilli were isolated in 55.3% of the samples, with concentrations of 3.33 ± 0.55 (log CFU/ml). The predominant species were L. plantarum (64%), L. fermentum (16%) and L. pentosus (9%). 28% of the isolated strains were resistant to gastric pH and bile salts, suggesting that they could be used as probiotics.

Se analizó la biodiversidad de especies de Lactobacillus en muestras de calostro de 116 madres chilenas mediante PCR y secuenciación del rDNA 16S. Se aislaron lactobacilos en 55,3% de las muestras, con concentraciones de 3,33 ± 0,55 (log UFC / ml). Las especies predominantes fueron L. plantarum (64%), L. fermentum (16%) y L. pentosus (9%). 28% de las cepas aisladas fueron resistentes a pH gástrico y a las sales biliares, lo que sugiere que podrían ser utilizados como probióticos.

Increase of plasma fatty acids without changes in n-6/n-3-PUFA ratio in asymptomatic obese subjects.

Obesity is associated with a low grade inflammation which contributes to the development of insulin resistance and diabetes. The aim of this study was to assess the total saturated (SFAs), monounsaturated (MUFAs) and polyunsaturated fatty acids (PUFAs) in plasma from asymptomatic obese subjects and to determine the arachidonic/eicosapentanoic acid ratio [ARA/EPA] as a marker of inflammation, and its eventual association with ultrasensitive CRP. Fourteen obese (34.4 +/- 11.1y.; BMI: 36.0 +/- 4,5 kg/m2) and 12 normal-weight (30.6 +/- 7.8y.; BMI: 23,6 +/- 2,4 kg/m2) subjects were recruited and their plasma fatty acids were determined by gas chromatography. usCRP was higher in the obese subjects (p = 0.01) and correlates with their body fat content. The percentages of SFAs, MUFAs, PUFAs were not affected in the obese subjects but their concentrations were increased, compared with the control group. However, no differences in the long chain PUFAs (DHA and EPA) concentrations or in the plasmatic ARA/EPA ratio were observed in these subjects. These observations do not support a relation between the ARA/EPA ratio and the presence of low grade inflammation evaluated by plasma usCRP in this group of asymptomatic obese subjects.

Saccharomyces boulardii in childhood.

INTRODUCTION: Probiotics are live microorganisms which confer a health benefit on the host. Saccharomyces boulardii, a yeast, has been found to be an effective probiotic in double-blind placebo-controlled randomized clinical studies. MATERIALS AND METHODS: We reviewed the established mechanisms of actions and clinical efficacy in children of S. boulardii. CONCLUSIONS: The mechanisms of action of S. boulardii depend mainly on the inhibition of some bacterial toxins, anti-inflammatory effects, and on stimulating effects on the intestinal mucosa such as trophic effects on the brush border enzymes and immunostimulatory effects. At present, in pediatric populations, there is evidence that S. boulardii is beneficial for the treatment of acute gastroenteritis and the prevention of antibiotic-associated diarrhea. More data are needed in other indications such as traveller's diarrhea, Helicobacter pylori eradication, and inflammatory bowel disease. S. boulardii is a yeast strain that has been extensively studied in vitro and in vivo. Recent data have opened the door for new therapeutic indications.

Urinary D-lactate excretion in infants receiving Lactobacillus johnsonii with formula.

BACKGROUND/AIMS: Supplementation with certain probiotics can improve gut microbial flora and immune function but should not have adverse effects. This study aimed to assess the risk of D-lactate accumulation and subsequent metabolic acidosis in infants fed on formula containing Lactobacillus johnsonii (La1). METHODS: In the framework of a double-blind, randomized controlled trial enrolling 71 infants aged 4-5 months, morning urine samples were collected before and 4 weeks after being fed formulas with or without La1 (1 x 10(8)/g powder) or being breastfed. Urinary D- and L-lactate concentrations were assayed by enzymatic, fluorimetric methods and excretion was normalized per mol creatinine. RESULTS: At baseline, no significant differences in urinary D-/L-lactate excretion among the formula-fed and breastfed groups were found. After 4 weeks, D-lactate excretion did not differ between the two formula groups, but was higher in both formula groups than in breastfed infants. In all infants receiving La1, urinary D-lactate concentrations remained within the concentration ranges of age-matched healthy infants which had been determined in an earlier study using the same analytical method. Urinary L-lactate also did not vary over time or among groups. CONCLUSIONS: Supplementation of La1 to formula did not affect urinary lactate excretion and there is no evidence of an increased risk of lactic acidosis.

FAO Technical meeting on prebiotics.

Recognizing the possible beneficial effect of prebiotics in food, the Food and Agriculture Organization of the United Nations (FAO) convened a Technical meeting to start work on the evaluation of the functional and health properties of prebiotics. A group of international experts agreed on guidelines, recommended criteria, and methodology for conducting a systematic approach for the evaluation of prebiotics leading to its safe use in food. It was recommended that a full expert consultation be convened under the auspices of FAO. This work provides governments, industry, and consumers with scientific advice in relation to functional and health aspects of prebiotics and general guidance for the assessment of prebiotics in relation to their nutritional properties or safety. These guidelines may also be used by Member Countries and Codex Alimentarius to identify and define what data need to be available to accurately substantiate health and nutrition claims.

Effect of a lactose-free milk formula supplemented with bifidobacteria and streptococci on the recovery from acute diarrhoea.

Probiotics have been proposed for the management and prevention of acute diarrhoea in infants. A double-blind, randomised, placebo controlled study was carried out in 224 Chinese infants 6 to 36 months of age with severe acute diarrhoea and free from moderate or severe malnutrition. After oral or parenteral rehydration, they were allocated to one of three groups: a lactose-free formula (Control); the same formula but with viable 10(8)CFU B. lactis Bb12 and 5x10(7)CFU St. thermophilus TH4 per gram of powder and, the same formula with the same microorganisms, but with 10(9)CFU/g and 5x10(8)CFU, respectively. Anthropometric parameters, duration of the diarrhoea and rotavirus shedding were evaluated. Eighty seven percent of the episodes were associated with rotavirus infection. The duration of the diarrhoea was not influenced by the intake of probiotics. However, a decrease of rotavirus shedding was observed in infants fed the formula with 10(9) Bb12/g, a finding of probable epidemiological importance in the transmission of this agent.

Iron, copper and immunocompetence.

Microminerals including copper and iron are essential to immunity and health in human beings. The development of powerful tools in analytical cell biology and molecular genetics has facilitated efforts to identify specific cellular and molecular functions of trace elements in the maturation, activation and functions of host defence mechanisms. Selected recent reports about the role of copper and iron nutrition on immune functions are critically analysed here. Effects of trace element supplementation on infectious morbidity are also reviewed. While micromineral deficiencies, in general, may have widespread effects on nearly all components of immune response, these effects can be reversed by supplementation. However, the conflicting effects of iron deficiency and iron supplementation in vitro on the defensive systems reveals the urgent need for further additional information on the in vivo situation. In the elderly, vaccination against respiratory infections is likely to protect only 30-70% of the population. However, it may be possible to modulate immune function and ultimately reduce the severity of infections through micronutrient supplementation. Thus, microminerals contribute to the maintenance of the balance between immunity and health in humans.

Effects of probiotic or prebiotic supplemented milk formulas on fecal microbiota composition of infants.

The aim of the study was to evaluate whether supplementation of milk-formulas with prebiotic fructo-oligosaccharides or a probiotic, Lactobacillus johnsonii La1 (La1), could modulate the composition of the fecal microbiota of formula-fed infants, compared to breastfed (BF) infants. Ninety infants close to 4 months of age were randomized into one of three groups to be blindly assigned to receive for 13 weeks: a) an infant formula (Control), b) the same formula with fructo-oligosaccharides (Prebio), or c) with La1 (Probio). At the end of this period, all infants received the control formula for 2 additional weeks. Twenty-six infants, breastfed throughout the study, were recruited to form group BF. Fecal samples were obtained upon enrolment and after 7 and 15 weeks. Bacterial populations were assessed with classical culture techniques and fluorescent in situ hybridisation (FISH). Seventy-six infants completed the study. On enrolment, higher counts of Bifidobacterium and Lactobacillus and lower counts of enterobacteria were observed in BF compared to the formula-fed infants; these differences tended to disappear at weeks 7 and 15. No major differences for Clostridium, Bacteroides or Enterococcus were observed between the groups or along the follow up. Probio increased fecal Lactobacillus counts (p<0.001); 88% of the infants in this group excreted live La1 in their stools at week 7 but only 17% at week 15. Increased Bifidobacterium counts were observed at week 7 in the 3 formula groups, similar to BF infants. These results confirm the presence of higher counts of bifidobacteria and lactobacilli in the microbiota of BF infants compared to formula-fed infants before dietary diversification, and that La1 survives in the infant digestive tract.

Effect of a milk formula with prebiotics on the intestinal microbiota of infants after an antibiotic treatment.

Antibiotics exert deleterious effects on the intestinal microbiota, favoring the emergence of opportunistic bacteria and diarrhea. Prebiotics are nondigestible food components that stimulate the growth of bifidobacteria. Our aim was to evaluate the effects on the intestinal microbiota of a prebiotic-supplemented milk formula after an antibiotic treatment. A randomized, double-blind, controlled clinical trial was carried out in 140 infants 1-2 y of age distributed into two groups after a 1-wk amoxicillin treatment (50 mg/kg/d) for acute bronchitis. The children received for 3 wk >500 mL/d of a formula with prebiotics (4.5 g/L) or a control without prebiotics. Fecal samples were obtained on d -7 (at the beginning of the antibiotic treatment), on d 0 (end of the treatment and before formula administration), and on d 7 and 21 (during formula administration). Counts of Bifidobacterium, Lactobacillus-Enterococcus, Clostridium lituseburiense cluster, Clostridium histolyticum cluster, Escherichia coli, and Bacteroides-Prevotella were evaluated by fluorescent in situ hybridization (FISH) and flow cytometry. Tolerance and gastrointestinal symptoms were recorded daily. Amoxicillin decreased total fecal bacteria and increased E. coli. The prebiotic significantly increased bifidobacteria from 8.17 +/- 1.46 on d 0 to 8.54 +/- 1.20 on d 7 compared with the control 8.22 +/- 1.24 on d 0 versus 7.95 +/- 1.54 on d 7. The Lactobacillus population showed a similar tendency while the other bacteria were unaffected. No gastrointestinal symptoms were detected during the prebiotic administration. Prebiotics in a milk formula increase fecal bifidobacteria early after amoxicillin treatment without inducing gastrointestinal symptoms.