Suckling Infant Research Articles

Acute decrease in mothers' cortisol following nursing and milk expression

The glucocorticoid hormone cortisol is an integral component of signaling pathways related to stress reactivity, energy balance, immune function, and other processes. In animal models, lactation is robustly associated with alterations in glucocorticoid signaling, and limited data suggest that similar changes may occur across human lactation. We asked whether milk letdown/secretion in breastfeeding mothers was associated with changes in cortisol, and whether such effects required presence of an infant. We measured changes in maternal salivary cortisol concentrations before and after nursing, the expression of breastmilk with an electric pump, or control activities. Participants conducted pre-session and post-session sampling (at 30 min) for all conditions, and provided a sample of pumped milk from one session. Both nursing and mechanical expression of breastmilk but not control were associated with equivalent declines in maternal cortisol concentration from pre-session values, indicating an effect of milk letdown on circulating cortisol independent of infant contact. Pre-session maternal salivary cortisol concentration was strongly and positively correlated with cortisol concentration in pumped milk samples, indicating that cortisol ingested by offspring provides a signal of maternal cortisol levels. Self-reported maternal stress was associated with higher pre-session cortisol concentrations, as well as with a larger drop in cortisol following nursing or pumping. These findings demonstrate that milk release—in the presence or absence of a suckling infant—regulates cortisol in mothers, and supports the potential for maternal signaling through breastmilk.

Postnatal exposure to organic pollutants in maternal milk in north-western Spain

Evaluation of postnatal exposure to organic pollutants is especially important for suckling infants during breastfeeding, a crucial perinatal growth period when organs and hormonal systems develop. We determined levels of 60 pollutants, including organochlorine pesticides (OCPs), organophosphorus pesticides (OPPs), pyrethroids (PYRs), polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and polybrominated diphenyl ethers (PBDEs), in 81 breast milk samples from breastfeeding mothers from Santiago de Compostela (north-western Spain). For most detected organic pollutants, levels were correlated with the season of milk sampling, maternal age at delivery, and place of residence. Dietary consumption habits (eggs, molluscs, and vegetable oils) were also correlated with OCP, OPP, PCB, PBDE and PYR levels. We also assessed the risk to infant health of exposure to organic pollutants in breast milk. PAHs, OCPs, OPPs, and PYRs accounted for almost 95% of the targeted organic pollutants in the samples analysed.

Human Breast Milk Contamination with Aflatoxins, Impact on Children’s Health, and Possible Control Means: A Review

Aflatoxins are natural toxicants produced mainly by species of the Aspergillus genus, which contaminate virtually all feeds and foods. Apart from their deleterious health effects on humans and animals, they can be secreted unmodified or carried over into the milk of lactating females, thereby posing health risks to suckling babies. Aflatoxin M1 (AFM1) is the major and most toxic aflatoxin type after aflatoxin B1 (AFB1). It contaminates human breast milk upon direct ingestion from dairy products or by carry-over from the parent molecule (AFB1), which is hydroxylated in the liver and possibly in the mammary glands by cytochrome oxidase enzymes and then excreted into breast milk as AFM1 during lactation via the mammary alveolar epithelial cells. This puts suckling infants and children fed on this milk at a high risk, especially that their detoxifying activities are still weak at this age essentially due to immature liver as the main organ responsible for the detoxification of xenobiotics. The occurrence of AFM1 at toxic levels in human breast milk and associated health conditions in nursing children is well documented, with developing countries being the most affected. Different studies have demonstrated that contamination of human breast milk with AFM1 represents a real public health issue, which should be promptly and properly addressed to reduce its incidence. To this end, different actions have been suggested, including a wider and proper implementation of regulatory measures, not only for breast milk but also for foods and feeds as the upstream sources for breast milk contamination with AFM1. The promotion of awareness of lactating mothers through the organization of training sessions and mass media disclosures before and after parturition is of a paramount importance for the success of any action. This is especially relevant that there are no possible control measures to ensure compliance of lactating mothers to specific regulatory measures, which can yet be appropriate for the expansion of breast milk banks in industrialized countries and emergence of breast milk sellers. This review attempted to revisit the public health issues raised by mother milk contamination with AFM1, which remains undermined despite the numerous relevant publications highlighting the needs to tackle its incidence as a protective measure for the children physical and mental health.

Transfer of Zolpidem to Cord Blood and Breast Milk: A Case Series Evaluating Zolpidem Serum Levels and Outcomes in Birth and Suckling Infants.

Background: Zolpidem is used for insomnia in pregnant and lactating women. Although zolpidem has been shown to cross the placenta and to be secreted into breast milk, it would not be expected to cause any adverse effects in newborn and breastfed infants. However, there is no relevant information on serum zolpidem levels in the newborn and breastfed infant from zolpidem-treated mother. This study aimed to present the outcomes of zolpidem exposure into infant who was delivered or breastfed by a zolpidem-treated mother. Methods: In this case series, zolpidem-treated pregnant women were recruited between September 2019 and April 2022, and maternal serum, cord blood, breast milk, and infants' serum were collected, and the zolpidem concentration in each sample was evaluated. Childbirth outcomes, including 1-month health care checkup, were also evaluated. Results: Three cases were recruited during investigation period. No spontaneous abortion or preterm live deliveries occurred. Oxygen intervention was required in one term infant, but the findings resolved on postpartum day 1. No medical intervention was required in other three infants. Zolpidem was not detected in infants' serum even after breastfeeding. There are no abnormal developmental findings in any of the infants in their 1-month health checkups. Conclusions: Zolpidem transferred into fetal circulation in utero and breast milk, however no harmful findings existed in infants during pregnancy and lactation. Exposure doses through breastfeeding is small, which may be a cause of rare detection from the infants' serum. Due to the limited number of cases, larger studies and integrated review are needed.

Quando non trascurare la splenomegalia nel lattante

Splenomegaly in children is really common and its main cause is infectious disease. The aetiology in newborns and older children is well established, while it is less defined in suckling infants. The paper describes three cases of splenomegaly in infants hospitalized in Oncohaematology Department of Sant’Orsola Malpighi Hospital. These infants presented with splenomegaly associated with other signs and symptoms that should prompt further investigations to exclude severe diseases. The aim of this paper is to offer paediatricians some indications for the diagnostic approach in infants with enlarged spleen. In particular, it reports the first level exams, the differential diagnosis and the red flags, the signs and symptoms that require special attention in infants with splenomegaly.

Colonic Handling of Calcium Appears To Occur Via Different Pathways in Mouse Models of Infants and Adults

ObjectivesCalcium (Ca2+) is a vital micronutrient for many physiological functions with the greatest rate of accumulation occurring during the critical period of infancy. Previous work has demonstrated that molecular mechanisms of intestinal Ca2+ absorption across the small intestine are significantly different in animal models of infants and adults to permit greater absorption early in life. The colon contributes to overall Ca2+ balance in adults via transcellular, TRPV6 mediated and paracellular claudin-2 and -12 mediated pathways. Whether these same colonic pathways contribute to overall Ca2+ absorption in infants is not known. Here we aimed to investigate the molecular details of Ca2+ absorption across the large intestine in murine models of infancy relative to older mice. MethodsMice at 14 days (P14) were used as a model of suckling infants and mice at 2 months were employed to represent adult physiology. Wildtype and mutant mice with a non-functioning Trpv6 or deletions of Cldn2 or Cldn12 were used. Net 45Ca2+ flux (JCa) and Ca2+ permeability (PCa) were measured in Ussing chambers. Gene expression was determined by real-time PCR. ResultsJCa indicates net absorption across the colon at both P14 and 2 months. While gene expression of Trpv6 and S100g suggest greater cellular uptake of Ca2+ into colonocytes at P14, net JCain vitro was not different than at 2 months. In contrast to previous work in mice at 2 months, TRPV6 does not mediate JCa at P14. PCa was 20% greater at P14 than 2 months, suggesting greater capacity for bidirectional diffusion of Ca2+ down an electrochemical gradient in younger mice. In contrast to previous work in mice at 2 months, claudin-2 and claudin-12 do not mediate PCa at P14 and, expression of Cldn2 and Cldn12 were significantly reduced in younger mice. ConclusionsThese results improve our understanding of intestinal Ca2+ handling during a critical age early in life. Future work is required to delineate molecular details under in vivo conditions of colonic Ca2+ transport in infants. Funding SourcesThis work was funded by grants from the Women and Children’s Health Research Institute, which is supported by the Stollery Children’s Hospital Foundation, and the National Sciences and Engineering Research Council to RTA, who is the Canada Research Chair in Renal Epithelial Transport Physiology.

Therapeutic monitoring of carbamazepine and its active metabolite during the 1st postnatal month: Influence of drug interactions

ObjectiveTo receive information about carbamazepine and its active metabolite 10,11-epoxide transport into mature milk and suckling infants. MethodsIn this cohort study, maternal serum, mature milk, and infant serum carbamazepine and epoxide levels were measured between the 6th and 29th postnatal day (carbamazepine in 1990–2017, epoxide in 1997–2017). Paired maternal serum, infant serum and milk levels were used for the assessment of ratios of this levels. The influence of combined treatment with enzyme-inducing antiepileptic drugs and valproic acid was assessed. Relationship between maternal serum, infant serum, and milk levels was also evaluated. ResultsMaternal carbamazepine levels were 1.4–10.4 mg/L, milk 0.5–6.7 mg/L and infant 0.5–2.6 mg/L. Maternal 10,11-epoxide levels were 0.3–5.4 mg/L, milk 0.3–3.7 mg/L and infant 0.3–0.6 mg/L. Highly significant correlations were observed exclusively between milk and maternal serum levels of both carbamazepine and 10,11-epoxide. Concomitant administration of enzyme-inducing antiepileptic drugs significantly increased the maternal apparent oral clearance of carbamazepine by approximately 130%. Carbamazepine combined with valproic acid significantly increased epoxide levels in milk and maternal serum but not in breastfed infants. ConclusionsIn breastfed infants, carbamazepine levels did not reach the lower limit of the therapeutic range used for the general epileptic population, and the majority of epoxide levels were less than the lower limit of quantification. Routine monitoring of carbamazepine in these infants is not compulsory. However, observation of breastfed infants is desirable. If signs of potential adverse reactions are evident, infant serum concentrations should be monitor.

Neonatal Suckling, Oxytocin, and Early Infant Attachment to the Mother.

The neuropeptide oxytocin (OT) promotes maternal care and social affiliation in adults but its importance in infant attachment still remains unknown. True animal models of infant attachment are extremely rare, and the sheep (in complement to non-human primates) is one of the few that provides the opportunity to investigate its neuroendocrinological basis. In the lamb, access to the udder has strong rewarding properties for the establishment of a preferential relationship with the mother. Therefore, the present study explored the possible involvement of OT through its release during close social contacts with the mother. The first experiment revealed that lambs having free access to the udder from birth onward developed, by 12 h of age, a clear preference for their mothers over another maternal ewe. Delaying access to the udder for six, four or even only 2 h starting at birth, by covering the ewe’s udder, resulted in the lack of such a preference without affecting general activity. These effects persisted in most cases at 24 h but by 72 h of age a bond with the mother was clearly expressed. Experiment two showed that social interactions with the mother were followed by a release of OT in the plasma when lambs had the possibility to suckle. Non-nutritive interactions were without effects. Preliminary data on two subjects suggested that OT might also increase in the cerebrospinal fluid after suckling. Finally, in the third experiment, oral administration of a non-peptide OT receptor antagonist (L-368-899, Merck) over the first 4 h after birth led to decreased exploration of the mother’s body compared to lambs receiving saline, and impaired the expression of a preference for the mother at 24 h. The effects were no longer observed at 48 h. Our findings demonstrate that both delayed access to the mother’s udder and OT receptor antagonist alter the onset of mother preference in newborn lambs. This suggests that central OT facilitates the development of filial attachment through its release during suckling.

Isolation and identification of milk oligosaccharide-degrading bacteria from the intestinal contents of suckling rats.

We report the isolation of bacteria capable of degrading milk oligosaccharides from suckling infant rats. The bacteria were successfully isolated via a selective enrichment method, in which the serially diluted intestinal contents of infant rats were individually incubated in an enrichment medium containing 3′-sialyllactose (3′-SL), followed by the isolation of candidate strains from streaked agar plates and selection of 3′-SL-degrading strains using thin-layer chromatography. Subsequent genomic and phenotypic analyses identified all strains as Enterococcus gallinarum. The strains were capable of degrading both 3′-SL and 6′-SL, which was not observed with the type strain of E. gallinarum used as a reference. Furthermore, a time-course study combining high-performance anion-exchange chromatography with pulsed amperometric detection revealed that the representative strain AH4 degraded 3′-SL completely to yield an equimolar amount of lactose and an approximately one-fourth equimolar amount of sialic acid after 24 hr of anaerobic incubation. These findings point to a possibility that the enterococci degrade rat milk oligosaccharides to “cross-feed” their degradants to other members of concomitant bacteria in the gut of the infant rat.

DOHaD: a disease-oriented, epoch-making, British-originated theory with Hungarian roots

On the basis of comparative epidemiological statistical studies, the ’developmental origin of health and disease’ (DOHaD) theory was published in 1986, testifying the interrelation between certain perinatal events, like under- and overfeeding as well as infant mortality with cardiovascular lethality in adults (Barker DJ, Osmond C, Lancet, 1986; 137: 1077–1081) – and at present it is widely extended. The theory is rather similar to the hormonal imprinting concept, which had been published 6 years earlier (Csaba G, Biol Rev Cambr Philos Soc. 1980; 55: 47–63 and Horm Metab Res. 1984; 16: 329–335). This demonstrated the role of perinatal encounter with hormones or hormone-like molecules with adult’s endocrino-pathological events based on animal experiments. Barker hypothesized the role of hormonal imprinting in DOHaD (Phillips DI, Barker DJ, Osmond C, Acta Endocrinol (Copenh). 1993; 129: 134–138: „A possible explanation is that thyroid hormones present in the breast milk and absorbed by the suckling infant could, by the process of hormonal imprinting, permanently down-regulate the set point of thyroid homeostasis”, and this could have encouraged him to create the theory. Both theories suggest the relationship between the perinatal events and adult-age disease manifestation, however, in the case of faulty imprinting, perinatal disease does not have a role in the provocation by imprinters (only the encounter between the imprinter and the hormone receptor), but in the case of DOHaD, this seems to be involved in the process. The whole process points to the disturbance of epigenetic programming. On the basis of the present standpoint, DOHaD is valid in non-communicative diseases, however, considering the impact of faulty hormonal imprinting to the immune system, the extension to communicative diseases is expected and likely also the involvement of lifespan. Further critical developmental period is the adolescence (puberty), when similar reprogramming could be possible and also in certain cases (e.g., in the immune system) disease-causing reprogramming could occur during the whole life. The two concepts are not racing, but using different methods for verification supplement and support each other, by building up identical conclusions (faulty reprogramming) giving epigenetical explanation for numerous diseases. DOHaD and its antecedent, hormonal imprinting are not only theories, but realities, which are commendable to consider in diagnosis and therapy. Studying the tendencies of human creativeness, in all probability, the importance of DOHaD (and faulty imprinting) will be growing in the near and far future. Orv Hetil. 2020; 161(16): 603–609.

Distinct expression patterns of aquaporin 3 and 5 in ductal and alveolar epithelial cells in mouse mammary glands before and after parturition.

Milk osmolarity maintains an isotonic status for suckling infants during lactation. However, it remains unclear how the water content in milk is regulated in lactating mammary glands. In lactating mammary alveoli and ducts, mammary epithelial cells (MECs) are in direct contact with milk. In this study, we focus on two types of water channels, aquaporin 3 (AQP3) and AQP5, in alveolar and ductal MECs before and after parturition. AQP3 showed diffuse localization in the cytoplasm of ductal MECs but concentrated localization in the basolateral membrane of alveolar MECs during the late pregnancy and lactation periods. Translocation of AQP5 from the cytoplasm toward the apical membrane occurred in ductal MECs immediately before parturition. Subsequently, we examined the hormonal influences on the expression of AQP3 and AQP5 in cultured MECs in vitro. Progesterone and estrogen distinctly increased AQP3 and AQP5 in cultured MECs, respectively. Cotreatment with prolactin and dexamethasone significantly decreased both AQP3 and AQP5. Prolactin also facilitated the translocation of AQP5 into the apical membrane of MECs. In cultured MECs, AQP3 was homogeneously expressed in MECs, whereas AQP5 showed different expression levels between MECs regardless of the hormonal treatment. Different activation states of the prolactin/STAT5 pathway were also observed between ductal and alveolar MECs. These findings suggest that the expression pattern of AQP3 and AQP5 is distinctly regulated by lactogenic hormones in alveolar and ductal MECs before and after parturition. AQP5 expressed in ductal MECs may function as a water channel to regulate milk osmolarity in mice.

Predictors of neonatal mortality: development and validation of prognostic models using prospective data from rural Bangladesh

ObjectiveTo assess the extent to which maternal histories of newborn danger signs independently or combined with birth weight and/or gestational age (GA) can capture and/or predict postsecond day (age>48 hours)...

Evaluation of Human Milk Microbiota by 16S rRNA Gene Next-Generation Sequencing (NGS) and Cultivation/MALDI-TOF Mass Spectrometry Identification.

The aim of the present study was to characterize human milk microbiota (HMM) with 16S rRNA gene amplicon next-generation sequencing and cultivation/matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF) mass spectrometry (MS) identification approaches. We analyzed 31 human milk samples from healthy Slovenian mothers. To check the accuracy of MALDI-TOF MS identification, several colonies representing most abundant genera and those, which could not be reliably identified by MALDI-TOF, were subjected to Sanger sequencing of their 16S rRNA gene. We showed that cultivation/MALDI-TOF MS was a suitable tool for culture-dependent determination of HMM. With both approaches, Staphylococcus and Streptococcus were found as predominant genera in HMM and the abundance of Staphylococcus was associated with decreased microbial diversity. In addition, we characterized factors that might influence HMM. The use of a breast pump was significantly associated with composition of HMM, higher microbial load, and lower abundance of cultivable staphylococci. Moreover, our study suggests that administration of probiotics to the suckling infant might influence HMM by increased abundance of lactobacilli and the presence of viable probiotic bacteria in human milk. However, since our study was observational with relatively small sample size, more targeted studies are needed to study possible transfer of probiotics to the mammary gland via an external route and the physiological relevance of these events.

Valproic acid concentrations in nursing mothers, mature milk, and breastfed infants in monotherapy and combination therapy

Valproic acid (VPA) is currently one of the four most often prescribed antiepileptic drugs (AEDs) in pregnancy. However, only a small number of studies have measured suckling infant serum levels of the drug. We studied the transport of VPA from breastfeeding mothers to the mature milk and breastfed infants and the influence of comedication with enzyme-inducing AEDs. The data of 30 nursing women treated by VPA were analyzed retrospectively. Mature milk, maternal, and infant serum levels were collected between the 6th and 32nd postnatal day and measured by gas chromatography during the years 1996–2017.Valproic acid levels varied from 5.4 to 69.0 mg/L (mean: 39.0 ± 16.1 mg/L) in the maternal serum, from <1.0 to 16.7 mg/L (mean: 1.6 ± 3.9 mg/L) in the milk, and from <1.0 to 17.5 mg/L (mean: 4.2 ± 4.3 mg/L) in the infant serum. The milk/maternal serum level ratio ranged from <0.03 to 0.25 (mean: 0.03 ± 0.06) and the infant/maternal serum level ratio from <0.03 to 0.61 (mean: 0.11 ± 0.13). Sixty-seven percent of milk and 33% of infant VPA concentrations were below the limit of quantification. No correlations were observed between maternal serum and milk levels or between maternal and infant serum levels. In conclusion, none of the milk or infant serum VPA levels reached the lower limit of the reference range used for the general population with epilepsy, so the degree of VPA exposure in breastfed infants is less than during gestation. Nevertheless, if signs of potential adverse reactions manifest, infant serum concentrations should be measured.

Dietary iron absorption during early postnatal life.

Inadequate iron levels during early life can have adverse consequences for the developing infant. Iron deficiency during this critical period of growth can affect brain development and cognitive function, problems that can be lifelong despite subsequent correction of the iron deficit. Therefore, it is critical that the suckling infant has sufficient iron for their developmental needs. Much of the iron used in the immediate post-natal period is stored iron that was acquired from the mother in the final trimester of pregnancy, however, despite having low iron levels, breast milk can also make a significant contribution to infant iron needs. This reflects the ability of the suckling infant to absorb dietary iron far more efficiently than is possible after weaning. The mechanisms underlying this enhanced iron absorption are poorly understood. The iron export protein ferroportin is essential for this process, as it is in adults, however, the role of other molecules normally involved in iron absorption following weaning is less clear. The composition and distribution of iron in breast milk may be important, as could the contribution of more distal parts of the gastrointestinal tract. This review discusses the potential role of each of the above components in intestinal iron absorption during suckling and highlights the need for further research into this important process.

Optimization and validation of a fast digestion method for the determination of major and trace elements in breast milk by ICP-MS

Breast milk guarantees all the nutrients required by infants during their first few months of life and remains the most important food source for their health and growth. However, the mother may transfer potentially toxic chemicals to the suckling infant through breastfeeding. The aim of this study was to optimize and validate a fast method for the determination of a total content of 34 elements (Al, As, B, Ba, Be, Bi, Ca, Cd, Co, Cr, Cs, Cu, Fe, K, Li, Mg, Mn, Mo, Na, Ni, P, Pb, Rb, Sb, Se, Si, Sn, Sr, Te, Ti, Tl, U, V, and Zn) in liquid and lyophilized breast milk. The samples were subjected to HNO3:H2O2 (2:1) digestion in an open vessel heated in a water bath (WBD; 80 °C) and subsequently analysed by quadrupole inductively coupled plasma mass spectrometry equipped with a collision-reaction interface.The performance of the proposed method was evaluated in terms of selectivity, detection and quantification limits, linearity, accuracy, and robustness by using standard reference materials and filed samples of breast milk. Compared to microwave-assisted acid digestion, the proposed open vessel digestion allows a significant reduction in treatment time and sample manipulation, while maintaining a similar analytical performance. Masses of 0.5 g of breast milk were efficiently digested with the WBD treatment allowing a residual carbon content lower than 60 mg L−1 and a residual acidity lower than 0.87 mol L−1 in final digested samples. Thus, it shows great potential for application to routine analysis. The method provides satisfactory detection limits and good performance (trueness and recovery percentages 80–111%; coefficient of variation <10%; and relative repeatability <15%) and allows a high sample throughput for multi-elemental determination in human biomonitoring studies.

Physiologically based pharmacokinetic (PBPK) modeling of human lactational transfer of methylmercury in China

Methylmercury can readily cross the human placental barrier and the blood-brain barrier and cause damage to the vulnerable developing brains of the fetus and infants. Most of the previous studies on the maternal transfer of methylmercury to the next generation have focused on the prenatal period. In this study, human physiologically based pharmacokinetic (PBPK) models of methylmercury were established for breastfeeding mothers and suckling infants based on the existing model prototypes of previous studies. Relevant parameters of the models were modified, and the validation was conducted based on measured data in North China. The models could effectively describe the human lactational transfer of methylmercury, including the time-dependent methylmercury levels in different tissues and organs of the breastfeeding mothers and suckling infants. The results indicated that 77.2% and 14.9% of methylmercury were excreted via hair and breast milk, respectively, from breastfeeding mothers during the first year after delivery. Meanwhile, 79.2% was excreted from the suckling infants during the first year after delivery via hair. Lactational transfer of methylmercury was considered an important pathway of methylmercury exposure for the breastfeeding infants, which accounted for approximately 80% of the accumulated adverse impacts at the early stages of human development.

Production of Chicken Egg Yolk Antibody (IgY) Against Recombinant Cholera Toxin B Subunit and Evaluation of Its Prophylaxis Potency in Mice.

Cholera toxin (CT), responsible for the harmful effects of cholera infection, is made up of one A subunit (enzymatic), and five B subunits (cell binding). The release of cholera toxin is the main reason for the debilitating loss of intestinal fluid. Inhibition of the B subunit (CTB) may block CT activity. To determine the effect of anti CTB-IgY against oral challenge with V. cholera in suckling infant mice. The binding domain of cholera toxin was amplified and ligated into pET28a vector. The pET28a (+)/ctb expression vector was confirmed by endonuclease digestion and sequence analysis. The expression of recombinant CTB in E. coli was performed by induction with IPTG. After immunizing the chickens with recombinant CTB, IgY was purified by water dilution method and NaCl precipitation and analyzed by SDS-PAGE. Moreover, the activity and specificity of the IgY antibody were assessed by ELISA. The SDS-PAGE and western blot techniques showed that CTB protein was successfully expressed and specifically recognized by polyclonal antibodies against the cholera toxin. The oral administration of anti- (V. cholera+CTB) in infant mice in challenge with active V. cholera bacterium demonstrated high rate of survival. The increase in the number of antibiotic resistant bacteria implies the necessity of finding novel antibiotics. Our results suggest the possibility of passive protection from purified IgY, hence implying that anti CTB-IgY may be useful in the treatment of cholera infections.

Mothers' Consumption of Soy Drink But Not Black Tea Increases the Flavonoid Content of Term Breast Milk: A Pilot Randomized, Controlled Intervention Study

Objective: We performed a pilot RCT to prove the hypothesis that a controlled ingestion of polyphenol-rich beverages (soy drink, decaffeinated black tea) in nutritive dosages by nursing women has an effect on the composition (flavonoid concentration, total antioxidant capacity) of breast milk. Methods: Healthy nursing women were supplemented with either 250 mL of a soy drink (12 mg isoflavones; n = 18), 300 mL decaffeinated black tea (67 mg catechins; n = 18), or 300 mL water (n = 8, control) for 6 days. Milk samples were collected before, during, and after intervention. Flavonoid content (isoflavones/catechins, HPLC) and total antioxidant capacity of milk and test drinks in milk specimens were assessed. Results: Isoflavone content (genistein and daidzein) in breast milk increased up to 12 nmol/L after soy drink consumption; the major flavonoids constituents of black tea (catechin, epicatechin, and respective conjugates) could not be detected in milk samples. With both interventions, the total antioxidant capacity of breast milk was not affected. Conclusions: Mothers' daily consumption of a soy drink considerably increases isoflavone content of breast milk resulting in an estimated daily exposure of 9.6 nmol isoflavones in a 4-month-old suckling infant. Luminal flavanol uptake from black tea consumed by the nursing mother may be too low to affect flavanol concentrations in breast milk.

Comparative analysis of native and permethylated human milk oligosaccharides by liquid chromatography coupled to high resolution mass spectrometry.

Human milk oligosaccharides (HMOs) represent the third most abundant components of milk after lactose and lipids. HMOs are indigestible by the suckling infant but can act as prebiotics and have significant biological functions regarding the organism defense against pathogens (such as bacteria or viruses) by preventing interactions with their receptors. Although constituted of only five distinct monosaccharide building blocks, HMOs are highly structurally diverse compounds with many co-existing structural isomers. Here we report the development and comparison of two distinct glycomic platforms based on liquid chromatography coupled to high resolution mass spectrometry (LC-MS) for analyzing HMOs. We have implemented and thoroughly compared the LC-MS of permethylated and native HMOs on reversed phase (RP) and porous graphitic carbon (PGC) columns for their ability to resolve the natural heterogeneity of milk oligosaccharides at the highest sensitivity. Our data essentially underlines the usefulness of analyzing HMOs as permethylated derivatives especially for getting more precise structural information at high sensitivity. For instance, permethylation annihilates gas-phase fucose migration during MS/MS experiments, thus facilitating spectra interpretation and giving access to relevant information regarding oligosaccharide branching and isomer distinction. At the opposite, LC-MS profiling of native HMOs (using PGC) in milk performed best in terms of detected species, while also being much faster in terms of sample preparation. Although less efficient than PGC chromatography, RPLC proved successful for separating pairs of permethylated isomeric HMOs. A key advantage of RP over PGC liquid chromatography is that retention times can be correlated to molecular weights, which can greatly facilitate further HMO identification using retention time prediction. Altogether these data lead us to think that LC-MS analysis of native HMOs (using PGC) can be used as first-line profiling approach while permethylation can be performed afterwards for facilitating structural characterization.