Breast Milk Microbiota Research Articles

Allergy development is associated with consumption of breastmilk with a reduced microbial richness in the first month of life.

Early colonization with a diverse microbiota seems to play a crucial role for appropriate immune maturation during childhood. Breastmilk microbiota is one important source of microbes for the infant, transferred together with maternal IgA antibodies. We previously observed that allergy development during childhood was associated with aberrant IgA responses to the gut microbiota already at 1month of age, when the IgA antibodies are predominantly maternally derived in breastfed infants. To determine the microbial composition and IgA-coated bacteria in breastmilk in relation to allergy development in children participating in an intervention trial with pre- and post-natal Lactobacillus reuteri supplementation. A combination of flow cytometric cell sorting and 16S rRNA gene sequencing was used to characterize the bacterial recognition patterns by IgA in breastmilk samples collected one month post-partum from 40 mothers whose children did or did not develop allergic and asthmatic symptoms during the first 7years of age. The milk fed to children developing allergic manifestations had significantly lower bacterial richness, when compared to the milk given to children that remained healthy. Probiotic treatment influenced the breastmilk microbiota composition. However, the proportions of IgA-coated bacteria, the total bacterial load and the patterns of IgA-coating were similar in breastmilk between mothers of healthy children and those developing allergies. Consumption of breastmilk with a reduced microbial richness in the first month of life may play an important role in allergy development during childhood.

Human Milk Microbiota: Transferring the Antibiotic Resistome to Infants.

Commensal bacterial population is believed to be a reservoir for antibiotic resistance genes (ARGs). The infant gut microbiota has relatively higher abundance of ARGs than the adults. These genes can get transferred from commensals to pathogens by horizontal gene transfer, which magnifies the spectrum of antibiotic resistance in the environment. The presence of ARGs in neo-nates and infants, with no prior antibiotic exposure, questions their origin in the naïve commensal population. Breast milk microbiota that is responsible for the initial seeding of infant gut microbiota has also been found to harbour a vast array of ARGs. This review discusses the recent findings that indicate the potential of breast milk microbiota to act as a vehicle for transmission of ARGs to infants.

Breast Milk Microbiota Is Shaped by Mode of Delivery and Intrapartum Antibiotic Exposure.

The mode of delivery has been suggested to modulate the bacterial composition of breast milk but the impact of intrapartum antibiotic use on the milk microbiota is currently not known. The aim of this study was to analyze the effects of the mode of the delivery and intrapartum antibiotic administration on the microbial composition of breast milk. Breast milk samples were collected from 84 healthy mothers 1 month after the delivery. In total, 61 mothers had delivered vaginally, 23 of which had received intrapartum antibiotics, 13 women had delivered with non-elective cesarean section, 7 of which had received antibiotics, and 10 mothers had delivered with elective cesarean section without intrapartum antibiotic treatment. Both mode of delivery and intrapartum antibiotic exposure were significantly associated with changes in the milk microbial composition as assessed by analysis of similarities (ANOSIM) test (p = 0.001). The mode of delivery had a more profound effect on the milk microbiota composition as compared to intrapartum antibiotic exposure. Although the clinical significance of breast milk microbiota is currently poorly understood, this study shows that cesarean section delivery has an independent effect on breast milk microbiota composition. The dysbiosis observed in infants born by cesarean section delivery may be aggravated by the aberrant breast milk microbiota.

Geographical location specific composition of cultured microbiota and Lactobacillus occurrence in human breast milk in China.

Breast milk bacteria play an important role in the early development of the gut microbiota and the immune system. Dominant living bacteria of 89 healthy Chinese women from 11 cities in five regions were analysed by broad-range yeast extract, casitone, and fatty acid and de Man, Rogosa, and Sharpe-based culturing coupled with 16S rRNA sequence and quantitative polymerase chain reaction. Principal coordinate analysis showed that human breast milk samples were classified into three groups, driven by Enterococcus (abundance in group 1, 63.13%), Streptococcus (abundance in group 2, 68.16%) and Staphylococcus (abundance in group 3, 55.17%). The microbiota profile was highly region-specific. Samples from the Northwest and North of China showed higher alpha diversity compared to other regions (p < 0.05). Staphylococcus, Streptococcus, and Enterococcus were the dominant genera in all samples. Lactobacillus had a high occurrence in samples from the Northwest and North, dominated by Lactobacillus reuteri and Lactobacillus gasseri. Samples of mothers with a high postpartum body mass index showed more Staphylococcus and less Lactobacillus and Streptococcus. Staphylococcus was negatively correlated with Lactobacillus and Streptococcus. The mode of delivery also affected the composition of microbiota, even after culture. These findings indicate differences between the North and South, provide effective information for collection of samples in which Lactobacillus is the predominant genus, and lower the detection limit for small amounts of bacteria.

938: Maternal IgA targets commensal microbiota in breast milk and the maternal and infant gut microbiomes

938: Maternal IgA targets commensal microbiota in breast milk and the maternal and infant gut microbiomes

HPV infection and bacterial microbiota in breast milk and infant oral mucosa.

ObjectiveWe investigated the association between bacterial microbiota in breast milk and the infant mouth. The influence of human papilloma virus (HPV) infection on infant oral microbiota was also assessed.Material and methodsAltogether 35 breast milk and 35 infant oral samples with known HPV status were selected from the Finnish Family HPV Study cohort. In total, there were 31 mother-infant pairs. The microbiota composition was characterized by 16S rRNA gene sequencing (V3-V4 region).ResultsHPV DNA was present in 8.6% (3/35) of the breast milk and 40% (14/35) of the infant oral samples. Eight shared genera between breast milk and infant oral were found; these included Streptococcus, Staphylococcus, Unclassified Gemellaceae, Rothia, Veillonella, Haemophilus, Propionibacterium and Corynebacterium. HPV status was not associated with either microbiota richness or diversity in the infant mouth. However, the infant oral microbiota clustered in different groups according to HPV status. We detected higher abundance of Veillonella dispar (p = 0.048) at species level in HPV negative infant oral samples. We did not detect differences in the breast milk microbiota composition related to HPV infection due to only three HPV positive milk samples.ConclusionsHPV infection is associated with distinct oral bacterial microbiota composition in infants. The direction of causality underlying the phenomenon remains unclear.

Mother's Milk: A Purposeful Contribution to the Development of the Infant Microbiota and Immunity.

Breast milk is the perfect nutrition for infants, a result of millions of years of evolution. In addition to providing a source of nutrition, breast milk contains a diverse array of microbiota and myriad biologically active components that are thought to guide the infant’s developing mucosal immune system. It is believed that bacteria from the mother’s intestine may translocate to breast milk and dynamically transfer to the infant. Such interplay between mother and her infant is a key to establishing a healthy infant intestinal microbiome. These intestinal bacteria protect against many respiratory and diarrheal illnesses, but are subject to environmental stresses such as antibiotic use. Orchestrating the development of the microbiota are the human milk oligosaccharides (HMOs), the synthesis of which are partially determined by the maternal genotype. HMOs are thought to play a role in preventing pathogenic bacterial adhesion though multiple mechanisms, while also providing nutrition for the microbiome. Extracellular vesicles (EVs), including exosomes, carry a diverse cargo, including mRNA, miRNA, and cytosolic and membrane-bound proteins, and are readily detectable in human breast milk. Strongly implicated in cell–cell signaling, EVs could therefore may play a further role in the development of the infant microbiome. This review considers the emerging role of breast milk microbiota, bioactive HMOs, and EVs in the establishment of the neonatal microbiome and the consequent potential for modulation of neonatal immune system development.

Methods and Strategies to Examine the Human Breastmilk Microbiome.

It has recently been discovered that breastmilk is not sterile, but contains a vast array of microbes, known collectively as the breastmilk microbiome. The breastmilk microbiome field is in its infancy, but over the last decade, our understanding of the microbial communities that inhabit the human body has increased exponentially, due in large part to novel next-generation sequencing technologies. These culture-independent, high-throughput molecular technologies have allowed biologists to investigate the entirety of microbiota present in breastmilk, which was previously poorly known. These approaches are novel and the methodologies surrounding the exploration of the breastmilk microbiota remain in flux. The objectives of this chapter are to outline what is known thus far and detail the optimal methods and strategies to conducting a breastmilk microbiome study from subject recruitment and milk collection to DNA extraction, high-throughput sequencing and bioinformatics analyses.

Breastfeeding-associated microbiota in human milk following supplementation with Lactobacillus rhamnosus GG, Lactobacillus acidophilus La-5, and Bifidobacterium animalis ssp. lactis Bb-12

Breastfeeding is one of the major factors affecting the early development of the infant gut microbiota, and weaning is associated with a shift in the gut microbiota toward a more adult composition. Through breastfeeding, infants receive bioactive components that shape their microbiota while also being exposed to the breast milk and breast surface microbial communities. Recent studies have suggested the possibility of an entero-mammary route of microbial transfer, opening the possibility of infant gut microbiota modulation through maternal probiotic supplementation. In this study, we have analyzed breast milk samples collected at 10 d and 3 mo postpartum from women participating in the Probiotics in the Prevention of Allergy among Children in Trondheim placebo controlled trial. Women who were randomized to the probiotic arm of the Probiotics in the Prevention of Allergy among Children in Trondheim trial received a fermented milk supplemented with Lactobacillus rhamnosus GG, Lactobacillus acidophilus La-5, and Bifidobacterium animalis ssp. lactis Bb-12, consuming this daily from 4 wk before their expected due date until 3 mo after birth. In total, 472 breast milk samples were assessed for the administered bacteria using quantitative real-time PCR and the microbiota transferred during breastfeeding was analyzed using 16S ribosomal RNA gene sequencing of 142 samples. We found that breastfeeding is unlikely to be a significant source of L. rhamnosus GG, L. acidophilus La-5, and B. animalis ssp. lactis Bb-12 for infants in the probiotic arm of the trial. Furthermore, maternal supplementation did not significantly affect the overall composition of the breast milk microbiota transferred during breastfeeding. We also present a descriptive analysis of this microbiota, which was largely dominated by Streptococcus and Staphylococcus genera at both 10 d and 3 mo postpartum. Samples collected at 3 mo postpartum had a statistically significant lower presence and relative abundance of the Staphylococcus genus. These samples also had a greater number of observed species and diversity, including more operational taxonomic units from the Rothia, Veillonella, Granulicatella, and Methylbacterium genera.

Bacterial Composition and Diversity in Breast Milk Samples from Mothers Living in Taiwan and Mainland China.

Human breast milk is widely recognized as the best source of nutrients for healthy growth and development of infants; it contains a diverse microbiota. Here, we characterized the diversity of the microbiota in the breast milk of East Asian women and assessed whether delivery mode influenced the microbiota in the milk of healthy breast-feeding mothers. We profiled the microbiota in breast milk samples collected from 133 healthy mothers in Taiwan and in six regions of mainland China (Central, East, North, Northeast, South, and Southwest China) by using 16S rRNA pyrosequencing. Lactation stage (months postpartum when the milk sample was collected) and maternal body mass index did not influence the breast milk microbiota. Bacterial composition at the family level differed significantly among samples from the seven geographical regions. The five most predominant bacterial families were Streptococcaceae (mean relative abundance: 24.4%), Pseudomonadaceae (14.0%), Staphylococcaceae (12.2%), Lactobacillaceae (6.2%), and Oxalobacteraceae (4.8%). The microbial profiles were classified into three clusters, driven by Staphylococcaceae (abundance in Cluster 1: 42.1%), Streptococcaceae (Cluster 2: 48.5%), or Pseudomonadaceae (Cluster 3: 26.5%). Microbial network analysis at the genus level revealed that the abundances of the Gram-positive Staphylococcus, Streptococcus, and Rothia were negatively correlated with those of the Gram-negative Acinetobacter, Bacteroides, Halomonas, Herbaspirillum, and Pseudomonas. Milk from mothers who had undergone Caesarian section (C-section group) had a significantly higher abundance of Lactobacillus (P < 0.05) and a higher number of unique unclassified operational taxonomic units (OTUs) (P < 0.001) than that from mothers who had undergone vaginal delivery (vaginal group). These findings revealed that (i) geographic differences in the microbial profiles were found in breast milk from mothers living in Taiwan and mainland China, (ii) the predominant bacterial families Streptococcaceae, Staphylococcaceae, and Pseudomonadaceae were key components for forming three respective clusters, and (iii) a significantly greater number of unique OTUs was found in the breast milk from mothers who had undergone C-section than from those who had delivered vaginally.

Human Breast Milk Microbiota is Influenced by Maternal Age and BMI, Stage of Lactation and Infant Feeding Practices

BackgroundHuman breast milk contains a diverse population of bacteria, but factors influencing the milk microbiota have not been described.ObjectiveWe explored if maternal age, BMI, stage of lactation and infant feeding practices influenced bacterial communities in breast milk of indigenous mothers in the western highlands of Guatemala.MethodsIn this cross‐sectional study, unilateral milk samples were collected from Mam‐Mayan women during early (5‐46d, n=33) or established (4‐6mo, n=43) lactation. Maternal age, BMI and feeding practices (exclusive, predominant and mixed feeding) were recorded. Milk bacterial communities were characterized by 16S ribosomal RNA amplicon sequencing using Illumina MiSeq platform. Sequence treatment, taxonomy assignment and operational taxonomic unit counts were performed with customized scripts based on Mothur and Dada2 MiSeq protocols. Taxonomy assignment was based on a &gt;95% confidence threshold. We compared normal (BMI 18.5–24.9) vs. overweight (BMI 25–30), early vs. established lactation and exclusive/predominant breastfeeding vs. mixed feeding. For all statistical tests, an effect size &gt;1.0, a confidence interval &gt;95% and a p‐value and q‐value &lt;0.05 were considered significant.ResultsHuman milk bacterial community was altered by maternal BMI, stage of lactation, infant feeding practices whereas bacterial diversity was only affected by maternal age. The highest bacterial diversity occurred in mothers aged 22–24 (Shannon index 1.8) and the lowest bacterial diversity occurred in mothers aged 13–15 (Shannon index 1). At the phylum level, stage of lactation modified the proportion of Firmicutes and Proteobacteria. Firmicutes was &gt;20% higher in early lactation and Proteobacteria was 30% higher in established lactation. A normal BMI was associated with higher proportions of Alphaproteobacteria and Betaproteobacteria at the class level. At the family level, exclusive/predominant breastfeeding when to compared to mixed feeding was associated with higher proportions of Corynebacteriaceae, Lactobacillaceae and Rhodobacteraceae.ConclusionHuman breast milk bacterial communities are determined by a complex interplay of maternal factors, stage of lactation and infant feeding practices. Exclusive breastfeeding might be necessary for the shaping of a healthy microbiota in the nursing infant. Finally, our data demonstrate the need to better understand the factors that contribute to the establishment of human breast milk microbiota, which could in turn influence infant growth and development.Support or Funding InformationNatural Sciences and Engineering Research Council of Canada (NSERC)

Breast Milk Polyamines and Microbiota Interactions: Impact of Mode of Delivery and Geographical Location

Background/Aims: The aim of the present study was to identify and quantify the polyamine levels in human milk obtained from different countries and through different modes of delivery, and to investigate their association with breast milk microbes. Methods: Mature breast milk samples were obtained from 78 healthy mothers after 1 month of lactation from 4 different geographical locations: Finland, Spain (Europe); South Africa (Africa); and China (Asia). Polyamines were determined using HPLC after dansyl derivatization and milk microbiota was obtained by 16S rRNA gene sequencing. Results: The mean values of polyamines in breast milk were 70.0, 424.2, and 610.0 nmol/dL for putrescine, spermidine and spermine, respectively, and 1,170.9 nmol/dL of total polyamines. The levels of putrescine were significantly higher in Spain (p < 0.05) and spermidine levels were significantly higher in Finland (p < 0.05) compared with other countries. Cesarean delivery had an impact on polyamine levels and it was related to an increase in the putrescine concentration being significant in Spanish samples (p < 0.01). Furthermore, putrescine levels were correlated positively with Gammaproteobacteria (r = 0.46, p < 0.001), especially with Pseudomonas fragi (r = 0.40, p < 0.001). Conclusions: The results demonstrate significant effect of geographical variations in human milk polyamine concentrations, being correlated with human milk microbiota composition. These differences may have an impact on infant development during lactation.

Mastitis in women: a new look at the old problem

Breast milk contains representatives of the microbiota which affect not only the health of child but also the condition of mammary glands in breastfeeding women both in the short and long term. In fact, dysbiosis of the breast milk microbiota caused by the mother’s body dysfunction as well as exposure to microbial and health factors, often contributes to the development of acute, subacute or subclinical mastitis. The pathology frequently results in the cessation of breastfeeding. Antibiotic resistance, ability to form biofilms, iron binding capacity, as well as the impact on the host’s immune response, are the common features of today’s bacterial pathogens (mainly staphylococci, streptococci and corynebacteria) which may also cause mastitis. This explains why people may become immune to antibiotic therapy, and why the development of new approaches in mastitis treatment using probiotics is becoming very attractive. Some strains of lactobacilli isolated from milk have already demonstrated high efficacy in the prevention and treatment of mastitis in lactating women.

Distinct Patterns in Human Milk Microbiota and Fatty Acid Profiles Across Specific Geographic Locations.

Breast feeding results in long term health benefits in the prevention of communicable and non-communicable diseases at both individual and population levels. Geographical location directly impacts the composition of breast milk including microbiota and lipids. The aim of this study was to investigate the influence of geographical location, i.e., Europe (Spain and Finland), Africa (South Africa), and Asia (China), on breast milk microbiota and lipid composition in samples obtained from healthy mothers after the 1 month of lactation. Altogether, 80 women (20 from each country) participated in the study, with equal number of women who delivered by vaginal or cesarean section from each country. Lipid composition particularly that of polyunsaturated fatty acids differed between the countries, with the highest amount of n-6 PUFA (25.6%) observed in the milk of Chinese women. Milk microbiota composition also differed significantly between the countries (p = 0.002). Among vaginally delivered women, Spanish women had highest amount of Bacteroidetes (mean relative abundance of 3.75) whereas Chinese women had highest amount of Actinobacteria (mean relative abundance 5.7). Women who had had a cesarean section had higher amount of Proteobacteria as observed in the milk of the Spanish and South African women. Interestingly, the Spanish and South African women had significantly higher bacterial genes mapped to lipid, amino acid and carbohydrate metabolism (p < 0.05). Association of the lipid profile with the microbiota revealed that monounsaturated fatty acids (MUFA) were negatively associated with Proteobacteria (r = -0.43, p < 0.05), while Lactobacillus genus was associated with MUFA (r = -0.23, p = 0.04). These findings reveal that the milk microbiota and lipid composition exhibit differences based on geographical locations in addition to the differences observed due to the mode of delivery.

Microbiota in Breast Milk of Chinese Lactating Mothers.

The microbiota of breast milk from Chinese lactating mothers at different stages of lactation was examined in the framework of a Maternal Infant Nutrition Growth (MING) study investigating the dietary habits and breast milk composition in Chinese urban mothers. We used microbiota profiling based on the sequencing of fragments of 16S rRNA gene and specific qPCR for bifidobacteria, lactobacilli and total bacteria to study microbiota of the entire breast milk collected using standard protocol without aseptic cleansing (n = 60), and the microbiota of the milk collected aseptically (n = 30). We have also investigated the impact of the delivery mode and the stage of lactation on the microbiota composition. The microbiota of breast milk was dominated by streptococci and staphylococci for both collection protocols and, in the case of standard collection protocol, Acinetobacter sp. While the predominance of streptococci and staphylococci was consistently reported previously for other populations, the abundance of Acinetobacter sp. was reported only once before in a study where milk collection was done without aseptic cleansing of the breast and rejection of foremilk. Higher bacterial counts were found in the milk collected using standard protocol. Bifidobacteria and lactobacilli were present in few samples with low abundance. We observed no effect of the stage of lactation or the delivery mode on microbiota composition. Methodological and geographical differences likely explain the variability in microbiota composition reported to date.

Administration of a multistrain probiotic product (VSL#3) to women in the perinatal period differentially affects breast milk beneficial microbiota in relation to mode of delivery

Probiotic supplementation to a mother during the perinatal period can have a positive impact on the breast milk composition. The aim of our study was to evaluate the effect of oral supplementation with the probiotic VSL#3, during late pregnancy and lactation, on breast milk levels of beneficial bacteria and some functional components (oligosaccharides and lactoferrin) potentially able to have a positive influence on the microbiota. Breast milk microbiota was analyzed by conventional and quantitative real-time PCR. In a double-blind, placebo-controlled, randomized trial, 66 women took daily either the probiotic (n=33) or a placebo (n=33). Intergroup analysis demonstrated that the amounts of both lactobacilli and bifidobacteria were significantly higher in the colostrum and mature milk of the mothers taking VSL#3 in comparison to those taking placebo. The analysis of bacterial strains and species present in breast milk of VSL#3 supplemented mothers indicated that the administered probiotic microorganisms did not pass from maternal gut to mammary gland. In women with vaginal delivery, significantly higher amounts of lactobacilli and bifidobacteria were detected in colostrum and mature milk of probiotic treated group in comparison to placebo group, whereas no significant difference was observed between groups in women who had caesarean section, neither in colostrum nor in mature milk. Milk levels of oligosaccharides and lactoferrin were similar in placebo and probiotic supplemented groups at all timepoints and regardless of the mode of delivery. Our results indicate a probiotic-dependent modulation of breast milk microbiota in vaginally delivering women, possibly exerted through a systemic effect.

Correlation between lactoferrin and beneficial microbiota in breast milk and infant's feces.

Lactoferrin (LF) is a natural component of human milk with antimicrobial, immunostimulatory and immunomodulatory properties. Several in vitro studies suggest that LF could promote an environment in the gut of neonates that favors colonization with beneficial bacteria. However, clinical studies on the correlation between the concentration of LF in breast milk and feces of infants and the gut microbiota in infants are lacking. In our study we analyzed the content of LF and the microbiota of breast milk and feces of infants of 48 mother-infant pairs (34 full-term and 14 pre-term infants) at birth and 30days after delivery. In the term group, a significant decrease of mean LF concentration between colostrum (7.0±5.1mg/ml) and mature milk (2.3±0.4mg/ml) was observed. In pre-term group, breast milk LF levels were similar to those observed in full-term group. Fecal LF concentration of healthy infants was extremely high both in term and pre-term infants, higher than the amount reported in healthy children and adults. In term infants mean fecal LF levels significantly increased from birth (994±1,828μg/ml) to 1month of age (3,052±4,323μg/ml). The amount of LF in the feces of 30day-old term infants was significantly associated with maternal mature milk LF concentration (p=0.030) confirming that breast milk represents the main source of LF found in the gut of infants. A linear positive correlation between colostrum and mature milk LF concentration was observed (p=0.008) indicating that milk LF levels reflect individual characteristics. In pre-term infants higher mean concentrations of fecal LF at birth (1,631±2,206μg/ml) and 30days after delivery (7,633±9,960μg/ml) were observed in comparison to full-term infants. The amount of fecal bifidobacteria and lactobacilli resulted associated with the concentration of fecal LF 3days after delivery (p=0.017 and p=0.026, respectively). These results suggest that high levels of fecal LF in neonates, particularly in the first days of life, could represent an important factor in the initiation, development and/or composition of the neonatal gut microbiota. Since early host-microbe interaction is a crucial component of healthy immune and metabolic programming, high levels of fecal LF in neonates may beneficially contribute to the immunologic maturation and well-being of the newborn, especially in pre-term infants.

Gut and Breast Milk Microbiota and Their Role in the Development of the Immune Function

The gastrointestinal immune system consists of both innate and adaptive immunity. The microbiota plays an important role in gastrointestinal immune function. In newborns, the maternal gut microbiota may either come across into the amniotic fluid or secrete factors that enter the amniotic fluid and affect the development of oral tolerance in utero. Adequate colonization must occur in the immediate postpartum period for the appropriate mucosal immune response since it is apparent that later attempts at colonization do not have the same impact. Human milk, being also a direct source of microbes, represents one of the main factors that play a critical role in influencing infant’s microbiota composition.

Human milk composition differs in healthy mothers and mothers with celiac disease

To investigate whether breast-milk composition and microbiota differ in healthy mothers and mothers with celiac disease (CD) to ultimately contribute to identify additional factors determining CD risk. Breast-milk samples from healthy mothers (n = 12) and mothers with CD (n = 12) were collected. Cytokines and secretory immunoglobulin A (sIgA) were analyzed by bead-arrays and flow cytometry and human milk oligosaccharides (HMOs) were assessed by capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection. Breast-milk microbiota composition was analyzed by conventional and quantitative real-time PCR. Breast milk from CD mothers showed significantly lower levels of interleukin (IL) 12p70 (P < 0.042), transforming growth factor (TGF)-β1 (P < 0.018) and sIgA (P < 0.003) and almost significantly lower levels of interferon (IFN)-γ (P < 0.058). Six mothers in each group belonged to the secretor Le(a-b+) type, one to the secretor Le(a-b-) type and five to the non-secretor Le(a+b-) type. CD mothers of non-secretor Le(a+b-) type showed increased Lacto-N-tetraose content (P < 0.042) compared with healthy mothers. CD mothers' milk showed reduced gene copy numbers of Bifidobacterium spp. (P < 0.026) and B. fragilis group (P < 0.044). CD mothers' breast milk is characterized by a reduced abundance of immunoprotective compounds (TGF-β1 and sIgA) and bifidobacteria. The reduction in these components could theoretically diminish the protective effects of breast-feeding on the child's future risk of developing CD.

Impact of lactation stage, gestational age and mode of delivery on breast milk microbiota

There is an increasing evidence of the immunological role of breast milk (BM) microbiota on infant health. This study aims to analyze several determining factors of milk microbiota. A total of 96 milk samples from 32 healthy mothers (19 preterm vs 13 at term gestations; and 15 vaginal deliveries vs 17 Cesarean sections) were longitudinally collected. Microbiota composition was studied by quantitative PCR and the influence of lactation stage, gestational age and delivery mode was evaluated. Globally, Lactobacillus, Streptococcus and Enterococcus spp. were the predominant bacterial groups. Total bacteria, Bifidobacterium and Enterococcus spp. counts increased throughout the lactation period. At all lactation stages, Bifidobacterium spp. concentration was significantly higher in milk samples from at term gestations than in preterm gestations. Higher bacterial concentrations in colostrum and transitional milk were found in Cesarean sections. Nevertheless, Bifidobacterium was detected more frequently in vaginal than in Cesarean deliveries. Lactation stage, gestational age and delivery mode all influence the composition of several bacteria inhabiting BM: Bifidobacterium, Lactobacillus, Staphylococcus, Streptococcus and Enterococcus spp., and, consequently, may affect the infant's early intestinal colonization.