Lacto-fermented foods such as sauerkraut have risen in marketing potential for local food processors; however, safety process parameters and established critical limits that validate process controls during production are limited. This study investigated process parameters such as vessel type (glass and plastic jars), salt concentration (0.0, 1.6, 2.4, 3.2, and 6.4%), use of starter culture, altered starting pH using lactic acid and acetic acid (to pH 5.50-5.00), and incubation temperature (22 degrees C or 30 degrees C) that may affect the fermentation conditions of sauerkraut over time. Samples prepared with 1.6, 2.4, and 3.2% salt significantly lowered the pH below 4.60 by day 5 (4.55 f 0.12), day 7 (4.60 f 0.11), and day 14 (4.45 f 0.16), respectively, compared to other concentrations. LAB counts significantly increased by at least 8-log for 1.6 and 2.4% salt sauerkraut after day 1.5. The pH levels decreased significantly for sauerkraut samples inoculated with starter culture after day 1 (4.55 f 0.23) compared to spontaneously fermented sauerkraut, in which pH decreased by day 2 (4.71 f 0.44). Faster pH drop was observed for sauerkraut samples fermented at 30 degrees C than those at 22 degrees C, as well as higher LAB counts (2-3.5 log difference). Using sauerkraut as a model system, these findings demonstrate how key fermentation parameters influence pH decrease, LAB growth, and spoilage outcomes, and provide technical guidance for establishing process controls in lacto-fermented foods. Additional validation may be required for applying specific controls to other products.

BACKGROUND:In low- and middle-income countries (LMICs), ∼250 million young children are at risk of not reaching their full growth and developmental potential. Early-life gut microbiota is increasingly recognized as a key factor influencing long-term growth and development, yet data from LMICs, including Zambia, remain limited. OBJECTIVES:This substudy, nested within a 2 × 2 cluster-randomized trial (NCT05120427) in Lusaka, Zambia, aimed to characterize early-life gut microbiota composition and diversity; examine associations with breastfeeding status, nutritional status, and maternal HIV status; and explore links between the microbiome and child growth and development. METHODS:Anthropometry and participant characteristics were assessed at baseline (2-11 mo) and after 18 mo of intervention; child development was measured at endline using the Global Scales for Early Development. Rectal swab stool samples were collected from a subsample of 247 infants at baseline and profiled via 16S rRNA gene sequencing. Microbial abundances were visualized with stacked bar plots and uniform manifold approximation and projection for dimensionality reduction. β-Diversity and its associations with explanatory variables were analyzed using distance-based redundancy analysis and nonmetric multidimensional scaling. Multivariate linear models evaluated genus-level associations with growth indicators, whereas random forest regression and partial least squares regression identified predictors of endline growth and development outcomes. RESULTS:We found that breastfeeding and nutritional status were significantly associated with microbiota composition. Mixed breastfeeding showed greater microbial diversity and broader taxa, whereas exclusive breastfeeding had higher Bifidobacterium abundance and increasing diversity with age. Despite these distinct trajectories, early microbiota features were weak predictors of later growth. CONCLUSIONS:Overall, further research is needed to clarify early microbiome drivers and their impact on child outcomes, particularly in LMIC settings. This trial was registered at https://clinicaltrials.gov/study/NCT05120427 as NCT05120427.

PURPOSE:To identify factors affecting macronutrient levels in human milk (HM) from mothers of preterm infants in Thailand by examining maternal diet and body mass index (BMI). BACKGROUND:HM fortification has become standard care to meet preterm infant's nutritional needs, but macronutrient content varies throughout lactation, leading to challenges in managing HM nutrients. Understanding maternal nutrition factors affecting HM macronutrients should be considered. METHOD:In this observational study, 47 mothers of premature infants were selected through convenience purposive sampling between October 2023 and March 2024. HM samples were collected at 2 time points: during 1-2 weeks of lactation (time point 1) and 3-4 weeks of lactation (time point 2) and analyzed using the Miris HM analyzer (Miris HMA™). Maternal dietary intake was assessed using 24-h dietary food recall records from 2 non-consecutive days and food frequency questionnaires (FFQs). Maternal BMI was measured using current body weight and height. RESULTS:Multiple linear regression analysis showed significant positive association between HM macronutrients and maternal dietary intake, including daily intake of carbohydrates, fats, and protein. No association was found between maternal BMI and HM macronutrients. CONCLUSIONS:This study supports that maternal dietary intake can affect the nutritional profile of HM. Monitoring and modifying maternal dietary intake during lactation may enhance macronutrient content of HM for preterm infants.

Bifidobacterium infantis are the primary colonizers of the infant gut, yet scientific research addressing the transmission of the genus Bifidobacterium to infants remains incomplete. This review examines microbial reservoirs of infant-type Bifidobacterium that potentially contribute to infant gut colonization. Accordingly, strain inheritance from mother to infant via the fecal-oral route is likely contingent on the bifidobacterial strain and phenotype, whereas transmission via the vaginal microbiota may be restricted to Bifidobacterium breve. Additional reservoirs include breastmilk, horizontal transfer from the environment, and potentially in utero transfer. Given that diet is a strong predictor of Bifidobacterium colonization in early life and the absence of Bifidobacterium is observed regardless of breastfeeding, it is likely that additional factors are responsible for bifidobacterial colonization early in life.

Background and Aims: Cannabigerol (CBG) is a nonintoxicating cannabinoid synthesized in the Cannabis sativa plant that is incorporated into dietary supplements. This study investigated the influence of dietary fat and an emulsified delivery vehicle on CBG pharmacokinetics (PKs) after oral ingestion by adults. Materials and Methods: Consented participants were enrolled in a double-crossover pilot study and were blinded to the delivery vehicle type (isolate or emulsification) and isocaloric meal condition (low-fat=<5 g fat/meal or high-fat [HF]=>30 g fat/meal). The concentration of CBG in human plasma was measured after a single 25 mg dose of CBG using liquid chromatography-tandem mass spectrometry (LC-MS/MS). PK parameters were calculated using noncompartmental analysis. Results: The PKs of the two delivery systems (emulsified vs. non-emulsified) were significantly impacted by the HF meal condition. Participants in the HF meal group exhibited significantly higher area under the plasma concentration time curve from time 0 to last quantifiable value, maximum concentration, and terminal half-life. Participants in the HF meal group also had a significantly lower terminal elimination rate constant and time to maximum concentration (T-max), in addition to decreased T-max variation. The threshold for bioequivalence between conditions was not met. An exploratory aim correlated anthropometric measures and previous day's dietary intake on PK parameters which yielded inconsistent results across dietary fat conditions. Conclusions: In aggregate, dietary fat had a greater effect on CBG PKs than the emulsified delivery vehicle. This supports accounting for dietary intake in development of therapeutics and administration guidelines for orally delivered CBG.

Human milk is universally recognized as the preferred food for infants during the first 6 mo of life because it provides not only essential and conditionally essential nutrients in necessary amounts but also other biologically active components that are instrumental in protecting, communicating important information to support, and promoting optimal development and growth in infants. Despite decades of research, however, the multifaceted impacts of human milk consumption on infant health are far from understood on a biological or physiological basis. Reasons for this lack of comprehensive knowledge of human milk functions are numerous, including the fact that milk components tend to be studied in isolation, although there is reason to believe that they interact. In addition, milk composition can vary greatly within an individual as well as within and among populations. The objective of this working group within the Breastmilk Ecology: Genesis of Infant Nutrition (BEGIN) Project was to provide an overview of human milk composition, factors impacting its variation, and how its components may function to coordinately nourish, protect, and communicate complex information to the recipient infant. Moreover, we discuss the ways whereby milk components might interact such that the benefits of an intact milk matrix are greater than the sum of its parts. We then apply several examples to illustrate how milk is better thought of as a biological system rather than a more simplistic "mixture" of independent components to synergistically support optimal infant health.

Human milk guides the structure and function of microbial commensal communities that colonize the nursing infant gut. Indigestible molecules dissolved in human milk establish a microbiome often dominated by bifidobacteria capable of utilizing these substrates. Interestingly, urea accounts for ~15% of total human milk nitrogen, representing a potential reservoir for microbiota that may be salvaged for critical metabolic operations during lactation and neonatal development. Accordingly, B. infantis strains are competent for urea nitrogen utilization, constituting a previously hypothetical phenotype in commensal bacteria hosted by humans. Urease gene expression, downstream nitrogen metabolic pathways, and enzymatic activity are induced during urea utilization to yield elevated ammonia concentrations. Moreover, biosynthetic networks relevant to infant nutrition and development are transcriptionally responsive to urea utilization including branched chain and other essential amino acids. Importantly, isotopically labeled urea nitrogen is broadly distributed throughout the expressed B. infantis proteome. This incisively demonstrates that the previously inaccessible urea nitrogen is incorporated into microbial products available for infant host utilization. In aggregate, B. infantis possesses the requisite phenotypic foundation to participate in human milk urea nitrogen recycling within its infant host and thus may be a key contributor to nitrogen homeostasis early in life.

Cannabidiol (CBD) is a non-intoxicating cannabinoid extracted from the cannabis plant that is used for medicinal purposes. Ingestion of CBD is claimed to address several pathologies, including gastrointestinal disorders, although limited evidence has been generated thus far to substantiate many of its health claims. Nevertheless, CBD usage as an over-the-counter treatment for gastrointestinal disorders is likely to expand in response to increasing commercial availability, permissive legal status, and acceptance by consumers. This systematic review critically evaluates the knowledge boundaries of the published research on CBD, intestinal motility, and intestinal motility disorders. Research on CBD and intestinal motility is currently limited but does support the safety and efficacy of CBD for several therapeutic applications, including seizure disorders, inflammatory responses, and upper gastrointestinal dysfunction (i.e., nausea and vomiting). CBD, therefore, may have therapeutic potential for addressing functional gastrointestinal disorders. The results of this review show promising in vitro and preclinical data supporting a role of CBD in intestinal motility. This includes improved gastrointestinal-related outcomes in murine models of colitis. These studies, however, vary by dose, delivery method, and CBD-extract composition. Clinical trials have yet to find a conclusive benefit of CBD on intestinal motility disorders, but these trials have been limited in scope. In addition, critical factors such as CBD dosing parameters have not yet been established. Further research will establish the efficacy of CBD in applications to address intestinal motility.

Bifidobacterium longum subsp. infantis (B. infantis) utilizes oligosaccharides secreted in human milk as a carbohydrate source. These human milk oligosaccharides (HMOs) integrate the nitrogenous residue N-acetylglucosamine (NAG), although HMO nitrogen utilization has not been described to date. Herein, we characterize the B. infantis nitrogen utilization phenotype on two NAG-containing HMO species, LNT and LNnT. This was characterized through in vitro growth kinetics, incorporation of isotopically labeled NAG nitrogen into the proteome, as well as modulation of intracellular 2-oxoglutarate levels while utilizing HMO nitrogen. Further support is provided by comparative transcriptomics and proteomics that identified global regulatory networks deployed during HMO nitrogen utilization. The aggregate data demonstrate that B. infantis strains utilize HMO nitrogen with the potential to significantly impact fundamental and clinical studies, as well as enable applications.

ScopeFucosylated human milk oligosaccharides (fHMOs) are metabolized by Bifidobacterium infantis and promote syntrophic interactions between microbiota that colonize the infant gut. The role of fHMO structure on syntrophic interactions and net microbiome function is not yet fully understood.Methods and resultsMetabolite production and microbial populations are tracked during mono‐ and co‐culture fermentations of 2ʹfucosyllactose (2ʹFL) and difucosyllactose (DFL) by two B. infantis strains and Eubacterium hallii. This is also conducted in an in vitro modeled microbiome supplemented by B. infantis and/or E. hallii. Metabolites are quantified by high performance liquid chromatography. Total B. infantis and E. hallii populations are quantified through qRT‐PCR and community composition through 16S amplicon sequencing. Differential metabolism of 2ʹFL and DFL by B. infantis strains gives rise to strain‐ and fHMO structure‐specific syntrophy with E. hallii. Within the modeled microbial community, fHMO structure does not strongly alter metabolite production in aggregate, potentially due to functional redundancy within the modeled community. In contrast, community composition is dependent on fHMO structure.ConclusionWhereas short chain fatty acid production is not significantly altered by the specific fHMO structure introduced to the modeled community, specific fHMO structure influences the composition of the gut microbiome.

Kombucha is a fermented tea beverage consumed for its probiotics and functional properties. It has a unique sensory profile driven by the properties of tea polyphenols and fermentation products, including organic acids. Fermentation temperature and sucrose content affect the fermentation process and the production of organic acids; yet less is known about their impacts on the sensory profile and consumer acceptance. Thus, we aimed to examine the impact of sucrose concentration and fermentation temperature on sensory attributes and liking. For this study, kombucha tea was fermented at three different concentrations of sucrose and fermented at two temperatures for 11 days. Fermentation was monitored by pH, brix, and titratable acidity, and consumers (n = 111) evaluated the kombucha for sensory attributes and overall liking. The fermentation temperature resulted in significant differences in titratable acidity, with higher temperatures producing more organic acids, resulting in higher astringency, and suppressed sweetness. The lower fermentation was reported as significantly more liked, with no difference in liking between the 7.5% and 10% sucrose kombucha samples. Fermentation temperature had the greatest impact on the sensory profile rather than sucrose concentration, which had a greater effect on the fermentation rate and production organic acids.

Asthma is associated with significant morbidity. The gut microbiome has been shown to effect asthma development and exacerbation. In this study, we tested an oral supplement containing Boswellia serrata (Indian frankincense) tree resin on allergic pulmonary inflammation and gut microbiome. An OVA based allergic airway model was used and mice were orally gavaged 100 mg/kg of Boswellia serrata as a supplementation throughout asthma sensitization and challenge. Treated mice showed significant weight loss, lower total lung leukocytes, eosinophil and Th2 cytokines, improved histology scoring and reduced reactivity to methacholine challenge. Asthmatic mice without Boswellia serrata supplementation showed a decrease in overall bacterial diversity, while treated mice were protected against loss. Boswellia serrata treated mice had a significant increase in Bifidobacterium, which was identified as Bifidobacterium pseudolongum. Oral administration of B. pseudolongum also reduced airway inflammation, suggesting Boswellia serrata may work as an anti-asthma agent via increases in B. pseudolongum from prebiotic influences.

The US government and many professional societies recommend that human milk be the sole source of nutrition for infants up to 6 months-of-age.1,2 However, only about 25% of infants in the US meet this goal signaling an urgent need for improved multi-level support for breastfeeding among societies, communities, and families.2 In addition, when breastfeeding is not possible, access to a safe and effective source of nutrition—provided by infant formula (IF) is critical. The search for bioactive ingredients found in human milk (HM) that could be added to formula has been intense among academic institutions, governmental research institutes, and industry.

Establishment of the gut microbiome during early life is a complex process with lasting implications for an individual's health. Several factors influence microbial assembly; however, breast-feeding is recognized as one of the most influential drivers of gut microbiome composition during infancy, with potential implications for function. Differences in gut microbial communities between breast-fed and formula-fed infants have been consistently observed and are hypothesized to partially mediate the relationships between breast-feeding and decreased risk for numerous communicable and noncommunicable diseases in early life. Despite decades of research on the gut microbiome of breast-fed infants, there are large scientific gaps in understanding how human milk has evolved to support microbial and immune development. This review will summarize the evidence on how breast-feeding broadly affects the composition and function of the early-life gut microbiome and discuss mechanisms by which specific human milk components shape intestinal bacterial colonization, succession, and function.

Plant-based foods contain bioactive compounds such as polyphenols that resist digestion and potentially benefit the host through interactions with their resident microbiota. Based on previous observations, we hypothesized that the probiotic Lactobacillus plantarum interacts with cranberry polyphenols and dietary oligosaccharides to synergistically impact its physiology. In this study, L. plantarum ATCC BAA-793 was grown on dietary oligosaccharides, including cranberry xyloglucans, fructooligosaccharides, and human milk oligosaccharides, in conjunction with proanthocyanidins (PACs) extracted from cranberries. As a result, L. plantarum exhibits a differential physiological response to cranberry PACs dependent on the carbohydrate source and polyphenol fraction introduced. Of the two PAC extracts evaluated, the PAC1 fraction contains higher concentrations of PACs and increased growth regardless of the oligosaccharide, whereas PAC2 positively modulates its growth during xyloglucan metabolism. Interestingly, fructooligosaccharides (FOS) are efficiently utilized in the presence of PAC1, as this L. plantarum strain does not utilize this substrate typically. Relative to glucose, oligosaccharide metabolism increases the ratio of secreted acetic acid to lactic acid. The PAC2 fraction differentially increases this ratio during cranberry xyloglucan fermentation compared with PAC1. The global transcriptome links the expression of putative polyphenol degradation genes and networks and metabolic phenotypes.

Biopolymer microgels were designed to encapsulate and protect probiotics during storage and passage through the upper human gastrointestinal tract (GIT), but then release them in the colon. An anaerobic probiotic strain (Bifidobacterium pseudocatenulatum G7; BPG7) was encapsulated within calcium alginate microgels. In some cases, either colloidal antacid (CaCO3) or colloidal antacid and nanoemulsion (nE) lipid droplets were co-encapsulated with the probiotics. These additives were used to control the porosity and/or internal pH of the microgels. Initially, the viability of free and encapsulated probiotics was evaluated when they were stored at 4 degrees C for 28 days. The free probiotics rapidly degraded during storage. Encapsulation greatly improved the stability of the probiotics, with the effectiveness depending on the formulation: antacid-microgels > control microgels > antacid-nE-microgels. These effects were mainly attributed to the ability of the colloidal CaCO3 particles to inhibit molecular diffusion processes. The ability of the microgels to protect the probiotics under simulated gastrointestinal tract (GIT) conditions was then assessed. The free probiotics were completely inactivated when exposed to gastric and small intestine conditions. In contrast, the antacid- and antacid-nE-microgels were highly effective at improving the viability of the probiotics during passage through the upper GIT. Moreover, these microgels were fermented under simulated colonic conditions, thereby releasing the probiotics. These results suggest that antacid-loaded calcium alginate microgels can be used for the encapsulation, protection, and colonic delivery of probiotics. Nevertheless, further studies are required using in vivo models.

BACKGROUND:White blood cell (WBC) DNA may contain methylation patterns that are associated with subsequent breast cancer risk. Using a high-throughput array and samples collected, on average, 1.3 years prior to diagnosis, a case-cohort analysis nested in the prospective Sister Study identified 250 individual CpG sites that were differentially methylated between breast cancer cases and noncases. We examined five of the top 40 CpG sites in a case-control study nested in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO) Cohort. METHODS:We investigated the associations between prediagnostic WBC DNA methylation in 297 breast cancer cases and 297 frequency-matched controls. Two WBC DNA specimens from each participant were used: a proximate sample collected 1 to 2.9 years and a distant sample collected 4.2-7.3 years prior to diagnosis in cases or the comparable timepoints in controls. WBC DNA methylation level was measured using targeted bisulfite amplification sequencing. We used logistic regression to obtain ORs and 95% confidence intervals (CI). RESULTS:A one-unit increase in percent methylation in ERCC1 in proximate WBC DNA was associated with increased breast cancer risk (adjusted OR = 1.29; 95% CI, 1.06-1.57). However, a one-unit increase in percent methylation in ERCC1 in distant WBC DNA was inversely associated with breast cancer risk (adjusted OR = 0.83; 95% CI, 0.69-0.98). None of the other ORs met the threshold for statistical significance. CONCLUSIONS:There was no convincing pattern between percent methylation in the five CpG sites and breast cancer risk. IMPACT:The link between prediagnostic WBC DNA methylation marks and breast cancer, if any, is poorly understood.

BACKGROUND:Preexisting immunity to SARS-CoV-2 could be related to cross-reactive antibodies to common human-coronaviruses (HCoVs). This study aimed to evaluate whether human milk antibodies against to S1 and S2 subunits SARS-CoV-2 are cross-reactive to S1 and S2 subunits HCoV-OC43 and HCoV-229E in mothers with a confirmed COVID-19 PCR test, in mothers with previous viral symptoms during COVID-19 pandemic, and in unexposed mothers; Methods: The levels of secretory IgA (SIgA)/IgA, secretory IgM (SIgM)/IgM, and IgG specific to S1 and S2 SARS-CoV-2, and reactive to S1 + S2 HCoV-OC43, and HCoV-229E were measured in milk from 7 mothers with a confirmed COVID-19 PCR test, 20 mothers with viral symptoms, and unexposed mothers (6 Ctl1-2018 and 16 Ctl2-2018) using ELISA; Results: The S2 SARS-CoV-2 IgG levels were higher in the COVID-19 PCR (p = 0.014) and viral symptom (p = 0.040) groups than in the Ctl1-2018 group. We detected a higher number of positive correlations between the antigens and secretory antibodies in the COVID-19 PCR group than in the viral symptom and Ctl-2018 groups. S1 + S2 HCoV-OC43-reactive IgG was higher in the COVID-19 group than in the control group (p = 0.002) but did not differ for the other antibodies; Conclusions: Mothers with a confirmed COVID-19 PCR and mothers with previous viral symptoms had preexisting human milk antibodies against S2 subunit SARS-CoV-2. Human milk IgG were more specific to S2 subunit SARS-CoV-2 than other antibodies, whereas SIgA and SIgM were polyreactive and cross-reactive to S1 or S2 subunit SARS-CoV-2.