Maternal breastfeeding is associated with offspring microbiome diversity; a secondary analysis of the MicrobeMom randomized control trial.

Background: Microbial dysbiosis in infancy can influence long-term health outcomes such as childhood obesity. The aim of this study is to explore relationships among maternal well-being during pregnancy, breastfeeding, and the infant gut microbiome. Methods: This is a secondary analysis of healthy pregnant women from the MicrobeMom study, a double-blind randomized control trial of maternal probiotic supplementation (Bifidobacterium breve 702258) versus placebo antenatally and up to 3 months postpartum. Maternal well-being was assessed using the WHO-5 well-being index at 16 weeks' and 34 weeks' gestation. Breastfeeding practices were recorded at discharge from hospital and at 1 month postpartum. Infant stool samples were obtained at 1 month of age. Next generation shotgun sequencing determined infant microbial diversity. Independent sample t-tests and Mann-Whitney U tests informed adjusted regression analysis, which was adjusted for delivery mode, antibiotics during delivery, maternal age and body mass index (BMI), and probiotic vs. control study group. Results: Women (n = 118) with at least one measure of well-being were on average 33 years (SD 3.93) of age and 25.09 kg/m2 (SD 3.28) BMI. Exclusive breastfeeding was initiated by 65% (n = 74). Any breastfeeding was continued by 69% (n = 81) after 1 month. In early and late pregnancy, 87% (n = 97/111) and 94% (n = 107/114) had high well-being scores. Well-being was not associated with infant microbial diversity at 1 month. In adjusted analysis, exclusive breastfeeding at discharge from hospital was associated with infant microbial beta diversity (PC2; 0.254, 95% CI 0.006, 0.038). At 1 month postpartum, any breastfeeding was associated with infant microbial alpha diversity (Shannon index; -0.241, 95% CI -0.498, -0.060) and observed species; (-0.325, 95% CI -0.307, -0.060), and infant microbial beta diversity (PC2; 0.319, 95% CI 0.013, 0.045). Exclusive breastfeeding at 1 month postpartum was associated with infant alpha diversity (Shannon index -0.364, 95% CI -0.573, -0.194; Simpson index 0.339, 95% CI 0.027, 0.091), and infant's number of observed microbial species (-0.271, 95% CI -0.172, -0.037). Conclusion: Breastfeeding practices at 1 month postpartum were associated with lower microbial diversity and observed species in infants at 1 month postpartum, which is potentially beneficial to allow greater abundance of Bifidobacterium. Clinical trial registration: ISRCTN53023014.

Detailed mapping of Bifidobacterium strain transmission from mother to infant via a dual culture-based and metagenomic approach.

A significant proportion of the infant gut microbiome is considered to be acquired from the mother during and after birth. Thus begins a lifelong and dynamic relationship with microbes that has an enduring impact on host health. Based on a cohort of 135 mother-infant (F = 72, M = 63) dyads (MicrobeMom: ISRCTN53023014), we investigated the phenomenon of microbial strain transfer, with a particular emphasis on the use of a combined metagenomic-culture-based approach to determine the frequency of strain transfer involving members of the genus Bifidobacterium, including species/strains present at low relative abundance. From the isolation and genome sequencing of over 449 bifidobacterial strains, we validate and augment metagenomics-based evidence to reveal strain transfer in almost 50% of dyads. Factors important in strain transfer include vaginal birth, spontaneous rupture of amniotic membranes, and avoidance of intrapartum antibiotics. Importantly, we reveal that several transfer events are uniquely detected employing either cultivation or metagenomic sequencing, highlighting the requirement for a dual approach to obtain an in-depth insight into this transfer process.

Ability of Bifidobacterium breve 702258 to transfer from mother to infant: the MicrobeMom randomized controlled trial.

BACKGROUND: The composition of the infant microbiome can have a variety of short- and long-term implications for health. It is unclear if maternal probiotic supplementation in pregnancy can affect the infant gut microbiome. OBJECTIVE: This study aimed to investigate if maternal supplementation of a formulation of Bifidobacterium breve 702258 from early pregnancy until 3 months postpartum could transfer to the infant gut. STUDY DESIGN: This was a double-blinded, placebo-controlled, randomized controlled trial of B breve 702258 (minimum 1 × 109 colony-forming units) or placebo taken orally from 16 weeks' gestation until 3 months postpartum in healthy pregnant women. The primary outcome was presence of the supplemented strain in infant stool up to 3 months of life, detected by at least 2 of 3 methods: strain-specific polymerase chain reaction, shotgun metagenomic sequencing, or genome sequencing of cultured B breve. A total of 120 individual infants' stool samples were required for 80% power to detect a difference in strain transfer between groups. Rates of detection were compared using the Fisher exact test. RESULTS: A total of 160 pregnant women with average age of 33.6 (3.9) years and mean body mass index of 24.3 (22.5-26.5) kg/m2, of whom 43% were nulliparous (n=58), were recruited from September 2016 to July 2019. Neonatal stool samples were obtained from 135 infants (65 in intervention and 70 in control group). The presence of the supplemented strain was detected through at least 2 methods (polymerase chain reaction and culture) in 2 infants in the intervention group (n=2/65; 3.1%) and none in the control group (n=0; 0%; P=.230). CONCLUSION: Direct mother-to-infant strain transfer of B breve 702258 occurred, albeit infrequently. This study highlights the potential for maternal supplementation to introduce microbial strains into the infant microbiome.

Interaction of replacing corn silage with soyhulls as a roughage source with or without 3% added wheat straw in the diet: impacts on intake, digestibility, and ruminal fermentation in steers fed high-concentrate diets.

Six ruminally cannulated steers [475.0 ± 49.6 kg initial body weight (BW)] were used in a 6 × 3 incomplete Latin square design (six treatments and three periods), to evaluate the impacts replacing of corn silage with pelleted soyhulls as roughage in high-concentrate finishing diets containing 30% modified distillers grains with solubles. Treatments were based on increasing dietary inclusion of soyhulls and consisted of: (1) Control (0), roughage supplied by dietary inclusion of 20% corn silage [dry matter (DM) basis]; (2) 50% replacement of corn silage with soyhulls (50); (3) 100% replacement of corn silage with soyhulls (100), and the same three treatments repeated with 3% added wheat straw (DM basis) replacing corn in the diet (0S, 50S, and 100S, respectively). Absolute dry matter intake (DMI; kg/d basis) tended to decrease both linearly and quadratically (P ≤ 0.09) and proportional DMI (% of BW) decreased linearly (P = 0.04) with increasing soyhull inclusion but was not affected by the addition of straw in the diet (P = 0.68). Total tract digestibility of organic matter and crude protein were not affected by soyhull inclusion or added straw (P ≥ 0.32). Ruminal pH did not differ (P = 0.65) with increasing soyhull inclusion but increased with the addition of straw (P < 0.01; 5.9 vs. 6.1 for no straw and straw, respectively). Molar proportions of acetate and butyrate decreased while propionate increased with increased soyhull inclusion (P ≤ 0.03; linearly and quadratically, respectively). Ruminal fluid kinetics were unaffected by either rate of replacement of corn silage with soyhulls or wheat straw inclusion (P ≥ 0.13). Decreases in DMI observed in this study would likely decrease finishing cattle performance and underscores the need for additional research before recommending this practice to cattle feeders.

Impacts of added roughage on growth performance, digestibility, ruminal fermentation, and ruminal pH of feedlot steers fed wheat-based feedlot diets containing 30% modified distillers grains with solubles.

Two experiments were conducted to evaluate the inclusion rate roughage in wheat-based diets containing modified distillers grains with solubles (MDGS) on feedlot performance (Feedlot Experiment), as well as digestibility, ruminal pH, and ruminal fermentation characteristics (Digestibility Experiment). The feedlot experiment utilized 72 Angus steers (392 ± 46.3 kg initial body weight) which were randomly assigned to 1 of 12 pens, 3 pens per treatment, to evaluate feedlot performance and carcass characteristics. Dietary treatments were 1) control; 10% roughage, 2) 12% roughage, 3) 14% roughage, and 4) 16% roughage. The digestibility experiment used four ruminally and duodenally cannulated steers (393 ± 33.0 kg) in a 4 × 4 Latin Square with either 10%, 12%, 14%, or 16% roughage as in the feedlot experiment. However, dietary roughage source was different between these two experiments and included a combination of grass hay and wheat straw (Feedlot Experiment), and corn silage (Digestibility Experiment). All data were analyzed with the Mixed Procedures of SAS. Feed intake was recorded, with duodenal and fecal output calculated using chromic oxide. Ruminal pH and fermentation were assessed. Growth performance and most carcass characteristics were not affected by increasing roughage (P ≥ 0.11). Marbling tended to decrease linearly (P = 0.10) with increasing roughage inclusion. Increasing dietary roughage content had no effect on organic matter intake (P = 0.60) in the digestibility experiment. Intake, duodenal flow, and digestibility of neutral detergent fiber and acid detergent fiber were not affected by treatment (P ≥ 0.16). Ruminal pH increased linearly (P < 0.01) as rate of roughage inclusion increased. Ruminal concentrations of acetate and butyrate increased, and propionate decreased in a linear fashion (P < 0.01) thereby increasing (P < 0.01) acetate and butyrate to propionate ratio with increasing dietary roughage. Our data indicate that increasing roughage inclusion in wheat-based diets including 30% MDGS increased ruminal pH and shifted ruminal fermentation patterns. Additionally, increasing roughage inclusion did not affect feedlot performance in steers fed wheat at 36% to 42% of dietary dry matter in combination with 30% MDGS.

Diet and mental health in pregnancy: Nutrients of importance based on large observational cohort data.

OBJECTIVE: To determine associations between dietary intake and well-being in pregnancy. METHODS: This retrospective cohort analysis combined three studies: the ROLO Study (a randomized controlled trial of a low-glycemic-index diet in pregnancy), the Pregnancy Exercise and nutrition Research Study with smartphone application support (PEARS), and a randomized controlled trial on probiotics. All data were collected before study interventions (16 wk). Dietary intakes during pregnancy were determined using 3-day food diaries. The five-item World Health Organization Well-Being Index was used to assess mental well-being. Initial associations were evaluated using Pearson correlations and further defined with multiple regression analysis adjusted for age, body mass index (BMI), Pobal Haase and Pratschke deprivation index, and metabolic equivalent of task minutes scores. RESULTS: A total of 1521 women were included in the analysis (mean age, 32+4 y; mean BMI, 27 kg/m2 [IQR, 17-56 kg/m2]). The mean well-being score was 59%. Regression analysis showed that fiber (B = 0.07; P = 0.02), magnesium (B = 0.08; P < 0.01), niacin (B = 0.09; P < 0.01), thiamine (B = 0.07; P = 0.01), and folate (B = 0.08; P = 0.02) were all positively and significantly associated with well-being in a pregnant population. The Benjamini-Hochberg procedure to correct for multiple testing was applied, and significance remained. CONCLUSIONS: Maternal nutrition and well-being are related during early pregnancy. Our findings suggest that fiber, magnesium, and particular B vitamins may be of importance for promoting positive mental well-being during pregnancy.

Impacts of bunk management on steer performance and ruminal hydrogen sulfide concentrations in steers fed modified distillers grains with solubles.

Two experiments were conducted to evaluate the impacts of bunk management on dry matter intake (DMI), growth performance, carcass characteristic, and hydrogen sulfide (H2S) concentrations in beef steers fed modified distillers grains with solubles (MDGS; DM basis). In Experiment 1, 139 steers (440.4 ± 31.0 kg) were randomly assigned to one of 16 pens with pen randomly assigned to one of two treatments: 1) Control (CON, bunks managed to be devoid of feed prior to feeding), or 2) Over-fed (OVF, bunks managed to have minimum of 2.54 cm of feed remaining each morning) during adaptation. Following adaptation all steers in Experiment 1 were transitioned to CON bunks and followed to finishing. In Experiment 2, 126 steers (445.4 ± 40.63 kg) were randomly assigned to one of 16 pens. Treatments in Experiment 2 were arranged in a 2 × 2 factorial and include the two bunk management strategies utilized in Experiment 1 (OVF or CON) and either 25% MDGS or 50% MDGS (DM basis). Ruminal H2S was measured via rumenocentesis during dietary adaptation. There were no differences (P ≥ 0.13) observed in either experiment for growth performance due to bunk management. In Experiment 1, OVF steers had greater (P = 0.001) DMI during adaptation; however, overall DMI was not different (P = 0.14) between treatments. In Experiment 2, DMI (d 0 to 104) tended to decrease (P = 0.09) with greater MDGS inclusion. Hot carcass weight, ribeye area, marbling score, and quality grade were not affected (P ≥ 0.48) by either bunk management or MDGS inclusion. In Experiment 2, back fat (1.30 vs. 1.17 ± 0.042 cm) and yield grade (3.2 vs. 3.0 ± 0.11) were greater (P = 0.03) for CON steers compared with OVF but were not affected (P = 0.59) by MDGS inclusion. In Experiment 1, H2S tended (P = 0.07) to be greater in steers on OVF compared with CON. In Experiment 2, bunk management strategy did not impact (P = 0.82) H2S concentrations. There was a MDGS inclusion × day interaction for H2S with steers fed 50% MDGS having greater (P < 0.01) H2S concentrations compared with steers fed 25% MDGS on d 28 and 35. Bunk management strategy during adaptation did not impact growth performance but did reduce intake in Experiment 1. Yield grade decreased when OVF bunk management was applied throughout Experiment 2. Response of H2S concentrations in the rumen were variable and likely influenced by inconsistencies in bunk management and resulting DMI during the early portions of the feedlot study.

Bacteremia in a Newborn with Hypocalcemic Seizures and Vitamin D Deficiency.

Infants with neonatal hypocalcemia often present with seizures, and neonatal hypocalcemia can be due to parathyroid (PTH) insufficiency or resistance. Causes of hypocalcemia with PTH elevation include increased phosphate load, vitamin D deficiency (VDD) or defects in metabolism, renal dysfunction, hypomagnesemia, genetic mutations resulting in end-organ resistance to PTH, or critical illness. Hypocalcemia has also been shown to be associated with Gram-negative bacteremia and sepsis in adults. We present the case of a full-term, formula-fed newborn presenting with late-onset hypocalcemic seizures and VDD in the setting of Klebsiella pneumoniae bacteremia. This case highlights that newborns presenting with hypocalcemic seizures should undergo a workup for sepsis.

Rubisco substitutions predicted to enhance crop performance through carbon uptake modelling.

Improving the performance of the CO2-fixing enzyme Rubisco is among the targets for increasing crop yields. Here, Earth system model (ESM) representations of canopy C3 and C4 photosynthesis were combined with species-specific Rubisco parameters to quantify the consequences of bioengineering foreign Rubiscos into C3 and C4 crops under field conditions. The 'two big leaf' (sunlit/shaded) model for canopy photosynthesis was used together with species-specific Rubisco kinetic parameters including maximum rate (Kcat), Michaelis-Menten constant for CO2 at ambient atmospheric O2 (Kc21%O2), specificity for CO2 to O2 (Sc/o), and associated heat activation (Ha) values. Canopy-scale consequences of replacing native Rubiscos in wheat, maize, and sugar beet with foreign enzymes from 27 species were modelled using data from Ameriflux and Fluxnet databases. Variation among the included Rubisco kinetics differentially affected modelled carbon uptake rates, and Rubiscos from several species of C4 grasses showed the greatest potential of >50% carbon uptake improvement in wheat, and >25% improvement in sugar beet and maize. This study also reaffirms the need for data on fully characterized Rubiscos from more species, and for better parameterization of 'Vcmax' and temperature response of 'Jmax' in ESMs.

Can a probiotic supplement in pregnancy result in transfer to the neonatal gut: A systematic review.

INTRODUCTION: The establishment of the neonatal gut microbiome is a crucial step that may have lifelong health implications. We aimed to systematically review evidence on maternal probiotic supplementation during pregnancy and vertical transfer of the corresponding strain to the infant gut. MATERIAL AND METHODS: Medline, CINAHL, Embase, Web of Science, and OVID were searched from inception to September 2018. Studies of maternal probiotic supplementation for a minimum duration of 2 weeks and analyses of neonatal stool samples were included. The primary outcome was presence of the specific probiotic strain in the infant stool. Electronic databases were searched for relevant studies and references were cross-checked. Risk of bias among included studies was assessed and data were extracted independently by two authors. RESULTS: Three studies were included in the review. Only one study was identified involving prenatal maternal probiotic supplementation alone. Neonatal colonization with the maternally administered probiotic was not demonstrated but supplementation with the probiotic influenced levels of a bacterial strain other than that found in the probiotic product. The other two studies identified included both prenatal and postnatal supplementation of either mother or infant. All three studies reported employing strain-specific isolation methodology to isolate the supplemented bacterial strain in infant stool but none used whole metagenome shotgun sequencing. CONCLUSIONS: Few studies investigating transfer of a specific probiotic bacterial strain from mother to infant were identified, showing inconclusive evidence of vertical transfer.

The association between the maternal diet and the maternal and infant gut microbiome: a systematic review.

During pregnancy, changes occur to influence the maternal gut microbiome, and potentially the fetal microbiome. Diet has been shown to impact the gut microbiome. Little research has been conducted examining diet during pregnancy with respect to the gut microbiome. To meet inclusion criteria, dietary analyses must have been conducted as part of the primary aim. The primary outcome was the composition of the gut microbiome (infant or maternal), as assessed using culture-independent sequencing techniques. This review identified seven studies for inclusion, five examining the maternal gut microbiome and two examining the fetal gut microbiome. Microbial data were attained through analysis of stool samples by 16S rRNA gene-based microbiota assessment. Studies found an association between the maternal diet and gut microbiome. High-fat diets (% fat of total energy), fat-soluble vitamins (mg/day) and fibre (g/day) were the most significant nutrients associated with the gut microbiota composition of both neonates and mothers. High-fat diets were significantly associated with a reduction in microbial diversity. High-fat diets may reduce microbial diversity, while fibre intake may be positively associated with microbial diversity. The results of this review must be interpreted with caution. The number of studies was low, and the risk of observational bias and heterogeneity across the studies must be considered. However, these results show promise for dietary intervention and microbial manipulation in order to favour an increase of health-associated taxa in the gut of the mother and her offspring.

Immobilisation of Delta-like 1 ligand for the scalable and controlled manufacture of hematopoietic progenitor cells in a stirred bioreactor.

BACKGROUND: Umbilical cord blood provides a source of hematopoietic stem cells for transplantation with immunological and availability advantages over conventional bone marrow sources. Limited cell numbers and slower engraftment from umbilical cord blood units has led to the clinical development of immobilised Notch ligand Delta-Like 1 to promote ex vivo expansion of a rapidly engrafting cell population. However, current immobilisation methods are not simple to scale in a controlled manner. RESULTS: Delta-Like 1 was immobilised onto streptavidin coated magnetic particles via a heterobifunctionalised polyethylene glycol linker molecule to provide an easily manipulated format of surface protein presentation. CD34+ enriched cord blood cells were treated with Delta-Like 1 immobilised particles, and immunophenotypic markers measured to monitor population distributions using cluster identification, characterization, and regression software. The amenability of the approach to scalability was evaluated in a micro-scale stirred tank bioreactor. Surface concentration of Delta-Like 1 was well controlled used differing stoichiometric reagent ratios. Protein immobilisation was a cost effective process and particles were efficiently removed from the final cell product. Immobilised Delta-Like 1 is functional and stimulates qualitatively similar CD34hi, CD38lo, CD90lo, CD133hi, CD135hi progenitor expansion in both static culture and scalable stirred culture platforms. CONCLUSIONS: Immobilised Delta-Like 1 in this form has the potential to improve the manufacturing efficiency and control of final ex vivo expanded cell product through compatibility with highly controlled and characterised suspension culture systems.

Immobilized hematopoietic growth factors onto magnetic particles offer a scalable strategy for cell therapy manufacturing in suspension cultures.

Hematopoietic therapies require high cell dosages and precise phenotype control for clinical success; scalable manufacturing processes therefore need to be economic and controllable, in particular with respect to culture medium and growth factor (GF) strategy. The aim of this work was to demonstrate the biological function, and integration within scalable systems, of a highly controllable immobilized growth factor (iGF) approach. GFs were biotinylated and attached to streptavidin coated magnetic particles. GF concentration during biotinylation, GF-biotin ratio, and GF lysine content were shown to control iGF surface concentration and enable predictable co-presentation of multiple GF on a single bead. Function was demonstrated for immobilized GMCSF, SCF, TPO and IL-3 in GF dependent cell lines TF-1 and M-07e. Immobilized GMCSF (iGMCSF) was analyzed to show sustained activity over 8 days of culture, a 2-3 order of magnitude potency increase relative to soluble factor, and retained functionality under agitation in a micro-scale stirred tank bioreactor. Further, short exposure to iGMCSF demonstrated prolonged growth response relative to soluble factor. This immobilization approach has the potential to reduce the manufacturing costs of scaled cell therapy products by reducing GF quantities and offers important process control opportunities through separation of GF treatments from the bulk media.

Clinician impression versus prescription drug monitoring program criteria in the assessment of drug-seeking behavior in the emergency department.

STUDY OBJECTIVE: We compare emergency provider impression of drug-seeking behavior with objective criteria from a state prescription drug monitoring program, assess change in opioid pain reliever prescribing after prescription drug monitoring program review, and examine clinical factors associated with suspected drug-seeking behavior. METHODS: This was a prospective observational study of emergency providers assessing a convenience sample of patients aged 18 to 64 years who presented to either of 2 academic medical centers with chief complaint of back pain, dental pain, or headache. Drug-seeking behavior was objectively defined as present when a patient had greater than or equal to 4 opioid prescriptions by greater than or equal to 4 providers in the 12 months before emergency department evaluation. Emergency providers completed data forms recording their impression of the likelihood of drug-seeking behavior, patient characteristics, and plan for prescribing pre- and post-prescription drug monitoring program review. Descriptive statistics were generated. We calculated agreement between emergency provider impression of drug-seeking behavior and prescription drug monitoring program definition, and sensitivity, specificity, and positive predictive value of emergency provider impression, using prescription drug monitoring program criteria as the criterion standard. A multivariate logistic regression analysis was conducted to determine clinical factors associated with drug-seeking behavior. RESULTS: Thirty-eight emergency providers with prescription drug monitoring program access participated. There were 544 patient visits entered into the study from June 2011 to January 2013. There was fair agreement between emergency provider impression of drug-seeking behavior and prescription drug monitoring program (κ=0.30). Emergency providers had sensitivity 63.2% (95% confidence interval [CI] 54.8% to 71.7%), specificity 72.7% (95% CI 68.4% to 77.0%), and positive predictive value 41.2% (95% CI 34.4% to 48.2%) for identifying drug-seeking behavior. After exposure to prescription drug monitoring program data, emergency providers changed plans to prescribe opioids at discharge in 9.5% of cases (95% CI 7.3% to 12.2%), with 6.5% of patients (n=35) receiving opioids not previously planned and 3.0% (n=16) no longer receiving opioids. Predictors for drug-seeking behavior by prescription drug monitoring program criteria were patient requests opioid medications by name (odds ratio [OR] 1.91; 95% CI 1.13 to 3.23), multiple visits for same complaint (OR 2.5; 95% CI 1.49 to 4.18), suspicious history (OR 1.88; 95% CI 1.1 to 3.19), symptoms out of proportion to examination (OR 1.83; 95% CI 1.1 to 3.03), and hospital site (OR 3.1; 95% CI 1.76 to 5.44). CONCLUSION: Emergency providers had fair agreement with objective criteria from the prescription drug monitoring program in suspecting drug-seeking behavior. Program review changed management plans in a small number of cases. Multiple clinical factors were predictive of drug-seeking behavior.