scholarly journals A Solution to Antifolate Resistance in Group B Streptococcus: Untargeted Metabolomics Identifies Human Milk Oligosaccharide-Induced Perturbations That Result in Potentiation of Trimethoprim

mBio ◽  
2020 ◽  
Vol 11 (2) ◽  
Author(s):  
Schuyler A. Chambers ◽  
Rebecca E. Moore ◽  
Kelly M. Craft ◽  
Harrison C. Thomas ◽  
Rishub Das ◽  
...  
Keyword(s):  
Human Milk ◽  
Metabolic Response ◽  
Group B Streptococcus ◽  
Resistance Mechanisms ◽  
Human Milk Oligosaccharides ◽  
Intrinsic Resistance ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Ultrahigh Performance Liquid Chromatography ◽  
Group B

ABSTRACT Adjuvants can be used to potentiate the function of antibiotics whose efficacy has been reduced by acquired or intrinsic resistance. In the present study, we discovered that human milk oligosaccharides (HMOs) sensitize strains of group B Streptococcus (GBS) to trimethoprim (TMP), an antibiotic to which GBS is intrinsically resistant. Reductions in the MIC of TMP reached as high as 512-fold across a diverse panel of isolates. To better understand HMOs’ mechanism of action, we characterized the metabolic response of GBS to HMO treatment using ultrahigh-performance liquid chromatography–high-resolution tandem mass spectrometry (UPLC-HRMS/MS) analysis. These data showed that when challenged by HMOs, GBS undergoes significant perturbations in metabolic pathways related to the biosynthesis and incorporation of macromolecules involved in membrane construction. This study represents reports the metabolic characterization of a cell that is perturbed by HMOs. IMPORTANCE Group B Streptococcus is an important human pathogen that causes serious infections during pregnancy which can lead to chorioamnionitis, funisitis, premature rupture of gestational membranes, preterm birth, neonatal sepsis, and death. GBS is evolving antimicrobial resistance mechanisms, and the work presented in this paper provides evidence that prebiotics such as human milk oligosaccharides can act as adjuvants to restore the utility of antibiotics.

scholarly journals Role of human milk oligosaccharides in Group B Streptococcus colonisation

2016 ◽  
Vol 5 (8) ◽  
pp. e99 ◽  
Author(s):  
Nicholas J Andreas ◽  
Asmaa Al-Khalidi ◽  
Mustapha Jaiteh ◽  
Edward Clarke ◽  
Matthew J Hyde ◽  
...  
Keyword(s):  
Human Milk ◽  
Group B Streptococcus ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Group B

scholarly journals Human Milk Oligosaccharides Reduce Murine Group B Streptococcus Vaginal Colonization with Minimal Impact on the Vaginal Microbiota

mSphere ◽  
2022 ◽  
Author(s):  
Marlyd E. Mejia ◽  
Samantha Ottinger ◽  
Alison Vrbanac ◽  
Priyanka Babu ◽  
Jacob J. Zulk ◽  
...  
Keyword(s):  
Preterm Birth ◽  
Human Milk ◽  
Group B Streptococcus ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Minimal Impact ◽  
Vaginal Colonization ◽  
Fetal Infection ◽  
Serious Disease ◽  
Group B

During pregnancy, GBS ascension into the uterus can cause fetal infection or preterm birth. In addition, GBS exposure during labor creates a risk of serious disease in the vulnerable newborn and mother postpartum.

scholarly journals Variation in Consumption of Human Milk Oligosaccharides by Infant Gut-Associated Strains of Bifidobacterium breve

2013 ◽  
Vol 79 (19) ◽  
pp. 6040-6049 ◽  
Author(s):  
Santiago Ruiz-Moyano ◽  
Sarah M. Totten ◽  
Daniel A. Garrido ◽  
Jennifer T. Smilowitz ◽  
J. Bruce German ◽  
...  
Keyword(s):  
Human Milk ◽  
Intestinal Microbiota ◽  
Glycosyl Hydrolase ◽  
Bifidobacterium Longum ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Bifidobacterium Breve ◽  
Infant Feces ◽  
Breast Fed

ABSTRACTHuman milk contains a high concentration of complex oligosaccharides that influence the composition of the intestinal microbiota in breast-fed infants. Previous studies have indicated that select species such asBifidobacterium longumsubsp.infantisandBifidobacterium bifidumcan utilize human milk oligosaccharides (HMO)in vitroas the sole carbon source, while the relatively fewB. longumsubsp.longumandBifidobacterium breveisolates tested appear less adapted to these substrates. Considering the high frequency at whichB. breveis isolated from breast-fed infant feces, we postulated that someB. brevestrains can more vigorously consume HMO and thus are enriched in the breast-fed infant gastrointestinal tract. To examine this, a number ofB. breveisolates from breast-fed infant feces were characterized for the presence of different glycosyl hydrolases that participate in HMO utilization, as well as by their ability to grow on HMO or specific HMO species such as lacto-N-tetraose (LNT) and fucosyllactose. AllB. brevestrains showed high levels of growth on LNT and lacto-N-neotetraose (LNnT), and, in general, growth on total HMO was moderate for most of the strains, with several strain differences. Growth and consumption of fucosylated HMO were strain dependent, mostly in isolates possessing a glycosyl hydrolase family 29 α-fucosidase. Glycoprofiling of the spent supernatant after HMO fermentation by select strains revealed that allB. brevestrains can utilize sialylated HMO to a certain extent, especially sialyl-lacto-N-tetraose. Interestingly, this specific oligosaccharide was depleted before neutral LNT by strain SC95. In aggregate, this work indicates that the HMO consumption phenotype inB. breveis variable; however, some strains display specific adaptations to these substrates, enabling more vigorous consumption of fucosylated and sialylated HMO. These results provide a rationale for the predominance of this species in breast-fed infant feces and contribute to a more accurate picture of the ecology of the developing infant intestinal microbiota.

scholarly journals Oligosaccharides Released from Milk Glycoproteins Are Selective Growth Substrates for Infant-Associated Bifidobacteria

2016 ◽  
Vol 82 (12) ◽  
pp. 3622-3630 ◽  
Author(s):  
Sercan Karav ◽  
Annabelle Le Parc ◽  
Juliana Maria Leite Nobrega de Moura Bell ◽  
Steven A. Frese ◽  
Nina Kirmiz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Human Milk ◽  
Bifidobacterium Longum ◽  
Bovine Milk ◽  
Selective Growth ◽  
Whey Protein Concentrate ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Growth Substrates

ABSTRACTMilk, in addition to nourishing the neonate, provides a range of complex glycans whose construction ensures a specific enrichment of key members of the gut microbiota in the nursing infant, a consortium known as the milk-oriented microbiome. Milk glycoproteins are thought to function similarly, as specific growth substrates for bifidobacteria common to the breast-fed infant gut. Recently, a cell wall-associated endo-β-N-acetylglucosaminidase (EndoBI-1) found in various infant-borne bifidobacteria was shown to remove a range of intactN-linked glycans. We hypothesized that these released oligosaccharide structures can serve as a sole source for the selective growth of bifidobacteria. We demonstrated that EndoBI-1 releasedN-glycans from concentrated bovine colostrum at the pilot scale. EndoBI-1-releasedN-glycans supported the rapid growth ofBifidobacterium longumsubsp.infantis(B. infantis), a species that grows well on human milk oligosaccharides, but did not support growth ofBifidobacterium animalissubsp.lactis(B. lactis), a species which does not. Conversely,B. infantisATCC 15697 did not grow on the deglycosylated milk protein fraction, clearly demonstrating that the glycan portion of milk glycoproteins provided the key substrate for growth. Mass spectrometry-based profiling revealed thatB. infantisconsumed 73% of neutral and 92% of sialylatedN-glycans, whileB. lactisdegraded only 11% of neutral and virtually no (<1%) sialylatedN-glycans. These results provide mechanistic support thatN-linked glycoproteins from milk serve as selective substrates for the enrichment of infant-associated bifidobacteria capable of carrying out the initial deglycosylation. Moreover, releasedN-glycans were better growth substrates than the intact milk glycoproteins, suggesting that EndoBI-1 cleavage is a key initial step in consumption of glycoproteins. Finally, the variety ofN-glycans released from bovine milk glycoproteins suggests that they may serve as novel prebiotic substrates with selective properties similar to those of human milk oligosaccharides.IMPORTANCEIt has been previously shown that glycoproteins serve as growth substrates for bifidobacteria. However, which part of a glycoprotein (glycans or polypeptides) is responsible for this function was not known. In this study, we used a novel enzyme to cleave conjugatedN-glycans from milk glycoproteins and tested their consumption by various bifidobacteria. The results showed that the glycans selectively stimulated the growth ofB. infantis, which is a key infant gut microbe. The selectivity of consumption of individualN-glycans was determined using advanced mass spectrometry (nano-liquid chromatography chip–quadrupole time of flight mass spectrometry [nano-LC-Chip-Q-TOF MS]) to reveal thatB. infantiscan consume the range of glycan structures released from whey protein concentrate.

scholarly journals Reconstruction of the Bifidobacterial Pan-Secretome Reveals the Network of Extracellular Interactions between Bifidobacteria and the Infant Gut

2018 ◽  
Vol 84 (16) ◽  
Author(s):  
Gabriele Andrea Lugli ◽  
Walter Mancino ◽  
Christian Milani ◽  
Sabrina Duranti ◽  
Francesca Turroni ◽  
...  
Keyword(s):  
Human Milk ◽  
Protein Translocation ◽  
Bifidobacterium Longum ◽  
Extracellular Proteins ◽  
Glycoside Hydrolases ◽  
Secreted Proteins ◽  
Human Milk Oligosaccharides ◽  
Secretion Systems ◽  
Milk Oligosaccharides ◽  
Content Type

ABSTRACT The repertoire of secreted proteins decoded by a microorganism represents proteins released from or associated with the cell surface. In gut commensals, such as bifidobacteria, these proteins are perceived to be functionally relevant, as they regulate the interaction with the gut environment. In the current study, we screened the predicted proteome of over 300 bifidobacterial strains among the currently recognized bifidobacterial species to generate a comprehensive database encompassing bifidobacterial extracellular proteins. A glycobiome analysis of this predicted bifidobacterial secretome revealed that a correlation exists between particular bifidobacterial species and their capability to hydrolyze human milk oligosaccharides (HMOs) and intestinal glycoconjugates, such as mucin. Furthermore, an exploration of metatranscriptomic data sets of the infant gut microbiota allowed the evaluation of the expression of bifidobacterial genes encoding extracellular proteins, represented by ABC transporter substrate-binding proteins and glycoside hydrolases enzymes involved in the degradation of human milk oligosaccharides and mucin. Overall, this study provides insights into how bifidobacteria interact with their natural yet highly complex environment, the infant gut.IMPORTANCE The ecological success of bifidobacteria relies on the activity of extracellular proteins that are involved in the metabolism of nutrients and the interaction with the environment. To date, information on secreted proteins encoded by bifidobacteria is incomplete and just related to few species. In this study, we reconstructed the bifidobacterial pan-secretome, revealing extracellular proteins that modulate the interaction of bifidobacteria with their natural environment. Furthermore, a survey of the secretion systems between bifidobacterial genomes allowed the identification of a conserved Sec-dependent secretion machinery in all the analyzed genomes and the Tat protein translocation system in the chromosomes of 23 strains belonging to Bifidobacterium longum subsp. longum and Bifidobacterium aesculapii.

scholarly journals Bifidobacterium longum subsp. infantis ATCC 15697 α-Fucosidases Are Active on Fucosylated Human Milk Oligosaccharides

2011 ◽  
Vol 78 (3) ◽  
pp. 795-803 ◽  
Author(s):  
David A. Sela ◽  
Daniel Garrido ◽  
Larry Lerno ◽  
Shuai Wu ◽  
Kemin Tan ◽  
...  
Keyword(s):  
Human Milk ◽  
Turnover Rate ◽  
Bifidobacterium Longum ◽  
Small Mass ◽  
Human Milk Oligosaccharides ◽  
High Turnover ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Biochemical Kinetics ◽  
Time Dependent Effect

ABSTRACTBifidobacterium longumsubsp.infantisATCC 15697 utilizes several small-mass neutral human milk oligosaccharides (HMOs), several of which are fucosylated. Whereas previous studies focused on endpoint consumption, a temporal glycan consumption profile revealed a time-dependent effect. Specifically, among preferred HMOs, tetraose was favored early in fermentation, with other oligosaccharides consumed slightly later. In order to utilize fucosylated oligosaccharides, ATCC 15697 possesses several fucosidases, implicating GH29 and GH95 α-l-fucosidases in a gene cluster dedicated to HMO metabolism. Evaluation of the biochemical kinetics demonstrated that ATCC 15697 expresses three fucosidases with a high turnover rate. Moreover, several ATCC 15697 fucosidases are active on the linkages inherent to the HMO molecule. Finally, the HMO cluster GH29 α-l-fucosidase possesses a crystal structure that is similar to previously characterized fucosidases.

scholarly journals Human Milk Oligosaccharides Promote the Growth of Staphylococci

2012 ◽  
Vol 78 (14) ◽  
pp. 4763-4770 ◽  
Author(s):  
K. M. Hunt ◽  
J. Preuss ◽  
C. Nissan ◽  
C. A. Davlin ◽  
J. E. Williams ◽  
...  
Keyword(s):  
Human Milk ◽  
Bacterial Communities ◽  
Bacterial Species ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Nutritional Conditions ◽  
Culture Independent ◽  
Lactating Mammary Gland

ABSTRACTHuman milk oligosaccharides (HMO), which constitute a major component of human milk, promote the growth of particular bacterial species in the infant's gastrointestinal tract. We hypothesized that HMO also interact with the bacterial communities present in human milk. To test this hypothesis, two experiments were conducted. First, milk samples were collected from healthy women (n= 16); culture-independent analysis of the bacterial communities was performed, HMO content was analyzed, and the relation between these factors was investigated. A positive correlation was observed between the relative abundance ofStaphylococcusand total HMO content (r= 0.66). In a follow-up study, we conducted a series ofin vitrogrowth curve experiments utilizingStaphylococcus aureusorStaphylococcus epidermidisand HMO isolated from human milk. HMO exhibited stimulatory effects on bacterial growth under various nutritional conditions. Analysis of culture supernatants from these experiments revealed that HMO did not measurably disappear from the culture medium, indicating that the growth-enhancing effects were not a result of bacterial metabolism of the HMO. Instead, stimulation of growth caused greater utilization of amino acids in minimal medium. Collectively, the data provide evidence that HMO may promote the growth ofStaphylococcusspecies in the lactating mammary gland.

scholarly journals Human milk oligosaccharides reduce murine group B Streptococcus vaginal colonization with minimal impact on the vaginal microbiota

2021 ◽  
Author(s):  
Marlyd E Mejia ◽  
Samantha Ottinger ◽  
Alison Vrbanac ◽  
Priyanka Babu ◽  
Jacob Zulk ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Human Milk ◽  
Biofilm Formation ◽  
Group B Streptococcus ◽  
Human Milk Oligosaccharides ◽  
Vaginal Colonization ◽  
Vaginal Epithelial Cells ◽  
Group B

Group B Streptococcus (GBS) colonizes the vaginal mucosa of a significant percentage of healthy women and is a leading cause of neonatal bacterial infections. Currently, pregnant women are screened in the last month of pregnancy and GBS-positive women are given antibiotics during parturition to prevent bacterial transmission to the neonate. Recently, human milk oligosaccharides (HMOs) isolated from breastmilk were found to inhibit GBS growth and biofilm formation in vitro, and women that make certain HMOs are less likely to be vaginally colonized with GBS. Using in vitro human vaginal epithelial cells and a murine vaginal colonization model, we tested the impact of HMO treatment on GBS burdens and the composition of the endogenous microbiota by 16S rRNA amplicon sequencing. HMO treatment reduced GBS vaginal burdens in vivo with minimal alterations to the vaginal microbiota. HMOs displayed potent inhibitory activity against GBS in vitro, but HMO pretreatment did not alter adherence of GBS or the probiotic Lactobacillus rhamnosus to human vaginal epithelial cells. Additionally, disruption of a putative GBS glycosyltransferase (Δsan_0913) rendered the bacterium largely resistant to HMO inhibition in vitro and in vivo but did not compromise its adherence, colonization, or biofilm formation in the absence of HMOs. We conclude that HMOs are a promising therapeutic bioactive to limit GBS vaginal colonization with minimal impacts on the vaginal microenvironment.

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