Fig 3 - available from: BMC Microbiology
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Sugar transporters identified in B. crudilactis (a) and B. mongoliense (b) genomes. Genes involved in milk sugars transport are indicated in red while the others are indicated in black. An E-value close to zero is indicated in dark yellow while an E-value far from zero is indicated in pale yellow. ORF: open reading frame; TCDB: Transporter classification database; E: E-value
Source publication
![Composition of the modified MRS2 media adapted from Tanimomo et al. [48]](publication/341230262/figure/tbl1/AS:888768143826947@1588910246895/Composition-of-the-modified-MRS2-media-adapted-from-Tanimomo-et-al-48_Q320.jpg)
Background:
Human milk oligosaccharides (HMO) could promote the growth of bifidobacteria, improving young children's health. In addition, fermentation of carbohydrates by bifidobacteria can result in the production of metabolites presenting an antivirulent activity against intestinal pathogens. Bovine milk oligosaccharides (BMO), structurally simi...
Contexts in source publication
Context 1
... were highlighted: one cytoplasmic β-hexosaminidase (GH20), one cytoplasmic α-L-fucosidase (GH95), two cytoplasmic β-galactosidases (GH2 and GH42) and two extracellular α-sialidases (GH33) (Fig. 2). In addition, 36 genes were predicted to be involved in sugars transport, among which 12 encode for HMO or BMO monosaccharides and oligosaccharides (Fig. 3b). Putative transporters belonging to the TCDB family 3.A.1.24, 3.A.1.25 and 3.A.1.2.20 provided glucose transport. Putative transporters 3.A.1.1.18, 3.A.1.2.22 and 3.A.1.1.48 assured transport of GlcNAc, oligosaccharides and lacto-N-biose or galacto-N-biose, ...
Context 2
... 19 genes encoding for GT and CBM were found, respectively (Fig. 1b and c). Two genes encoding for cytoplasmic β-galactosidases (GH2 and GH42) were highlighted (Fig. 2). From the B. mongoliense DSM 21395 genome, 28 genes were predicted to be involved in sugars transport, among which eight encode for HMO or BMO monosaccharides and oligosaccharides (Fig. 3b). Putative transporters belonging to the TCDB family 2.A.1.68.1 and 2.A.7.5.5 provided glucose transport. Putative transporters 2.A.2.2.3 and 3.A.1.1.48 assured transport of galactose and lacto-N-biose or galacto-N-biose, ...
Context 3
... to the schematic representation of the BMO metabolism available in the Additional file 3: Figure S3, B. mongoliense could be able to digest most of the oligosaccharides found in BMO, except the sialic acid, which is supposed to stay in the external environment. In the same way, B. mongoliense could be able to metabolise 3′SL (NeuAcα2-3Galβ1-4Glc) using an extracellular α-sialidase. ...
Context 4
... highlights the putative capacity of B. mongoliense to metabolise the 3′SL and other complex BMO. The carbohydrate residues issuing from this degradation, such as sialic acid, glc, gal or GlcNac, could be involved in this antivirulent effect ( Figure S3). The potential metabolites from 3′SL or whey degradation are probably different from those mentioned above and could explain why the 3′SL and whey fermentation by B. mongoliense FR/49/f/2 CFSM did not have the same antivirulence effect on E. coli O157:H7 compared to B. crudilactis FR/62/b/3 CFSM. ...
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Citations
... HMOs have bactericidal, bacteriostatic, and antibiofilm properties [7]. Existence of α-and β-bonds in HMOs makes them resistant to the upper gut acidic environment and are poorly digested owing to lack of indigenous glycosidases [8]. Thus, they are mostly found as such in later parts of the intestine where they feed commensal gut microbial flora. ...
Human milk oligosaccharides (HMOs) are a crucial ingredient in mother’s milk for infant health. These are a structurally diverse group of soluble bioactive glycans having antiviral and anti-bacterial properties and promote the growth of probiotics, particularly Bifidobacteria. They modulate the immune system and are important for infant’s brain development. Not all infants are fortunate enough to receive mother milk, and rely on industrially produced infant formulas which lack HMOs. Thus, synthesizing HMOs through different methods is becoming more appealing form an industrial perspective. Despite huge efforts to obtain predominant HMOs through chemical, chemo-enzymatic, enzymatic, and whole-cell biotransformation of microorganisms, a limited number of HMOs could be produced in sufficient quantities due to lack of canonical enzymes that can add different metabolic pathways together to produce several HMO. Particularly, lacto-N-neotetraose and 2′-fucosyllactose are being added to infant formula and are well-tolerated by infants. Amide other methods, enzymatic and whole-cell biotransformation of microorganisms are promising approaches, but it requires intervention of innovative methodologies of synthetic biology. This review highlights innovative approaches, such as using plant system, and employing synthetic biology including redesigning of enzyme structure for producing HMOs at the industrial level. An interesting approach for synthesizing whole milk components in mammary cell lines is also mentioned.
... A previous investigation of the resident microbiome of 128 Italian raw milk cheeses revealed that B. mongoliense is the most prevalent bifidobacterial species (8). Neverthe less, to date, the genetic information of the B. mongoliense taxon is based on a rather limited number of genotypes (14,15). Consequently, to further investigate the genetic variability of this species, a cultivation-dependent strategy was employed, allowing the isolation of 10 novel B. mongoliense strains from different raw milk cheeses and cheesemaking sites (Table 1). ...
... Overall, qPCR evaluation of B. mongoliense BMONG18 revealed that this taxon is able to utilize 3′-SL and mucin-based medium for growth (Fig. 4). In this context, it has already been demonstrated that B. mongoliense is able to metabolize most oligosaccharides present in bovine milk, including 3′-SL, due to an extracellular α-sialidase (14). Among all possible combinations with bifidobacterial strains, the quantity of BMONG18 cells was shown to increase up to threefold in those bi-associations that include B. longum PRL2022 or B. adolescentis PRL2023 cells when cultivated on MRS supplemented with all different carbohydrates compared to their corresponding mono-associations (Fig. 4). ...
The microbial ecology of raw milk cheeses is determined by bacteria originating from milk and milk-producing animals. Recently, it has been shown that members of the Bifidobacterium mongoliense species may become transmitted along the Parmigiano Reggiano cheese production chain and ultimately may colonize the consumer intestine. However, there is a lack of knowledge regarding the molecular mechanisms that mediate the interaction between B. mongoliense and the human gut. Based on 128 raw milk cheeses collected from different Italian regions, we isolated and characterized 10 B. mongoliense strains. Comparative genomics allowed us to unveil the presence of enzymes required for the degradation of sialylated host-glycans in B. mongoliense, corroborating the appreciable growth on de Man-Rogosa-Sharpe (MRS) medium supplemented with 3’-sialyllactose (3′-SL) or 6’-sialyllactose (6′-SL). The B. mongoliense BMONG18 was chosen, due to its superior ability to utilize 3′-SL and mucin as representative strain, to investigate its behavior when co-inoculated with other bifidobacterial species. Conversely, members of other bifidobacterial species did not appear to benefit from the presence of BMONG18, highlighting a competitive scenario for nutrient acquisition. Transcriptomic data of BMONG18 reveal no significant differences in gene expression when cultivated in a gut simulating medium (GSM), regardless of whether cheese was included or not. Furthermore, BMONG18 was shown to exhibit high adhesion capabilities to HT29-MTX human cells, in line with its colonization ability of a human host.
IMPORTANCE
Fermented foods are nourishments produced through controlled microbial growth that play an essential role in worldwide human nutrition. Research interest in fermented foods has increased since the 80s, driven by growing awareness of their potential health benefits beyond mere nutritional content. Bifidobacterium mongoliense, previously identified throughout the production process of Parmigiano Reggiano cheese, was found to be capable of establishing itself in the intestines of its consumers. Our study underscores molecular mechanisms through which this bifidobacterial species, derived from food, interacts with the host and other gut microbiota members.
... It is a key component in the human milk microbiota, which can modulate newborn immune development [39]. In cow's milk, Bifidobacterium species are also present and able to degrade bovine milk oligosaccharides [40]. In our canine study, while observed in both colostrum and milk, Bifidobacterium prevalence (37.5 and 62.5% respectively) and abundance (0.2 ± 0.3 and 0.1 ± 0.3%, respectively) were very low. ...
Background
A mother’s milk is considered the gold standard of nutrition in neonates and is a source of cytokines, immunoglobulins, growth factors, and other important components, yet little is known about the components of canine milk, specifically colostrum, and the knowledge related to its microbial and metabolic profiles is particularly underwhelming. In this study, we characterized canine colostrum and milk microbiota and metabolome for several breeds of dogs and examined profile shifts as milk matures in the first 8 days post-whelping.
Results
Through untargeted metabolomics, we identified 63 named metabolites that were significantly differentially abundant between days 1 and 8 of lactation. Surprisingly, the microbial compositions of the colostrum and milk, characterized using 16S rRNA gene sequencing, were largely similar, with only two differentiating genera. The shifts observed, mainly increases in several sugars and amino sugars over time and shifts in amino acid metabolites, align with shifts observed in human milk samples and track with puppy development.
Conclusion
Like human milk, canine milk composition is dynamic, and shifts are well correlated with developing puppies’ needs. Such a study of the metabolic profile of canine milk, and its relation to the microbial community, provides insights into the changing needs of the neonate, as well as the ideal nutrition profile for optimal functionality. This information will add to the existing knowledge base of canine milk composition with the prospect of creating a quality, tailored milk substitute or supplement for puppies.
... Additionally, this genus presented small but noticeable amounts in MC4 and MC5 cheese samples with 2.23% and 2.48%. Previously, this genus with probiotic potential was detected in cheeses made with similar production methods, such as Camembert, Brie, and Reblochon cheeses (Bondue et al., 2020). To the best of our knowledge, there is no report on the presence of Bifidobacterium in Turkish moldripened cheeses. ...
Kuflu cheese, a popular variety of traditional Turkish mold-ripened cheeses, is characterized by its semi-hard texture and blue-green color. It is important to elucidate the microbiota of Kuflu cheese produced from raw milk to standardize and sustain its sensory properties. This study aimed to examine the bacteria, yeasts, and filamentous mold communities in Kuflu cheese using high-throughput amplicon sequencing based on 16S and ITS2 regions. Lactococcus, Streptococcus, and Staphylococcus were the most dominant bacterial genera while Bifidobacterium genus was found to be remarkably high in some Kuflu cheese samples. Penicillium genus dominated the filamentous mold biota while the yeasts with the highest relative abundances were detected as Debaryomyces, Pichia, and Candida. The genera Virgibacillus and Paraliobacillus, which were not previously reported for mold-ripened cheeses, were detected at high relative abundances in some Kuflu cheese samples. None of the genera that include important food pathogens like Salmonella, Campylobacter, Listeria were detected in the samples. This is the first experiment in which the microbiota of Kuflu cheeses were evaluated with a metagenomic approach. This study provided an opportunity to evaluate Kuflu cheese, which was previously examined for fungal composition, in terms of both pathogenic and beneficial bacteria.
... However, most bifidobacteria cannot sustain the presence of oxygen, which makes their industrial application problematic. Nevertheless, some strains of B. mongoliense and B. crudilactis can tolerate the presence of oxygen and a low pH, providing the potential use of these species as probiotics [54]. A MAG affiliated to Brachybacterium nesterenkovii was identified in the metagenome of suluguni-like cheese (03SU). ...
Fermented milk products (FMPs) contain probiotics that are live bacteria considered to be beneficial to human health due to the production of various bioactive molecules. In this study, nine artisanal FMPs (kefir, ayran, khurunga, shubat, two cottage cheeses, bryndza, khuruud and suluguni-like cheese) from different regions of Russia were characterized using metagenomics. A metagenomic sequencing of ayran, khurunga, shubat, khuruud and suluguni-like cheese was performed for the first time. The taxonomic profiling of metagenomic reads revealed that Lactococcus species, such as Lc. lactis and Lc. cremoris prevailed in khuruud, bryndza, one sample of cottage cheese and khurunga. The latter one together with suluguni-like cheese microbiome was dominated by bacteria, affiliated to Lactobacillus helveticus (32–35%). In addition, a high proportion of sequences belonging to the genera Lactobacillus, Lactococcus and Streptococcus but not classified at the species level were found in the suluguni-like cheese. Lactobacillus delbrueckii, as well as Streptococcus thermophilus constituted the majority in another cottage cheese, kefir and ayran metagenomes. The microbiome of shubat, produced from camel’s milk, was significantly distinctive, and Lentilactobacillus kefiri, Lactobacillus kefiranofaciens and Bifidobacterium mongoliense represented the dominant components (42, 7.4 and 5.6%, respectively). In total, 78 metagenome-assembled genomes with a completeness ≥ 50.2% and a contamination ≤ 8.5% were recovered: 61 genomes were assigned to the Enterococcaceae, Lactobacillaceae and Streptococcaceae families (the Lactobacillales order within Firmicutes), 4 to Bifidobacteriaceae (the Actinobacteriota phylum) and 2 to Acetobacteraceae (the Proteobacteria phylum). A metagenomic analysis revealed numerous genes, from 161 to 1301 in different products, encoding glycoside hydrolases and glycosyltransferases predicted to participate in lactose, alpha-glucans and peptidoglycan hydrolysis as well as exopolysaccharides synthesis. A large number of secondary metabolite biosynthetic gene clusters, such as lanthipeptides, unclassified bacteriocins, nonribosomal peptides and polyketide synthases were also detected. Finally, the genes involved in the synthesis of bioactive compounds like β-lactones, terpenes and furans, nontypical for fermented milk products, were also found. The metagenomes of kefir, ayran and shubat was shown to contain either no or a very low count of antibiotic resistance genes. Altogether, our results show that traditional indigenous fermented products are a promising source of novel probiotic bacteria with beneficial properties for medical and food industries.
... are widely used as probiotic microorganisms. Bifidobacterium mongoliense strains showing in vitro resistance to gastric and pancreatic juices, and bile salts, and having ability to digest milk oligosaccharides and produce antivirulent metabolites, have been reported [53,55]. Therefore, efforts should be made in the future to recover and investigate the probiotic potential of bifidobacteria from IBT and IT cheeses. ...
High-throughput DNA sequencing (HTS) was used to study the microbial diversity of commercial traditional Izmir Tulum (IT) and Izmir Brined Tulum (IBT) cheeses from Izmir, Türkiye. Simultaneously, cultivation-dependent methods were used to isolate, identify and characterize bacterial strains displaying probiotic potential. At the phylum level, Firmicutes dominated the microbiota of both cheese types comprising >98% of the population. Thirty genera were observed, with Streptococcus being the most abundant genus and with Streptococcus thermophilus and S. infantarius subsp. infantarius being the most abundant species. Genera, including Bifidobacterium and Chryseobacterium, not previously associated with IT and IBT, were detected. IT cheeses displayed higher operational taxonomic units (OTUs; Richness) and diversity index (Simpson) than IBT cheeses; however, the difference between the diversity of the microbiota of IT and IBT cheese samples was not significant. Three Lacticaseibacillus paracasei strains isolated from IBT cheeses exhibited probiotic characteristics, which included capacity to survive under in vitro simulated gastrointestinal conditions, resistance to bile salts and potential to adhere to HT-29 human intestinal cells. These findings demonstrate that Tulum cheeses harbor bacterial genera not previously reported in this cheese and that some strains display probiotic characteristics.
... Interestingly, in human milk, Bi dobacterium is a central bacterial player, known to metabolize human milk oligosaccharides, a key component in human milk, known, amongst other things, to modulate newborn immune development [39]. In cow's milk Bi dobacterium species are also present and able to degrade bovine milk oligosaccharides [40]. In our canine study, while observed in both colostrum and milk, Bi dobacterium prevalence (37.5 and 62.5% respectively) and abundance (0.2 ± 0.3 and 0.1 ± 0.3%, respectively) were very low. ...
... The available supernatant volume(10,25,40,50, 80, or 100 µL) was collected and moved to a separate tube. Cold aqueous methanol (80%) was added to the supernatant, and each sample was immediately vortexed for 30 s after the addition of methanol. ...
Background
A mother’s milk is considered the gold standard of nutrition in neonates and is a source of cytokines, immunoglobulins, growth factors, and other important components, yet little is known about the components of canine milk, specifically colostrum, and the knowledge related to its microbial and metabolic profiles is particularly underwhelming. In this study we characterized canine colostrum and milk microbiota and metabolome of several breeds of dogs and examined profile shifts as milk matures in the first 8 days post-whelping.
Results
Through untargeted metabolomics, we identified 63 named metabolites that were significantly differentially expressed between days 1 and 8 of lactation, whereas the microbial compositions of the colostrum and milk, characterized using 16S rRNA gene sequencing, were, surprisingly, largely similar, with only two differentiating genera. The shifts observed, mainly increases in several sugars and amino sugars over time and shifts in amino acid metabolites, align with shifts observed in human milk samples and track with puppy development.
Conclusion
Like human milk, canine milk composition is dynamic, and shifts are well correlated with developing puppies’ needs. Such a study of the metabolic profile of canine milk, and its relation to the microbial community, provides insights into the changing needs of the neonate, as well as the ideal profile for optimal functionality. This information will add to the existing knowledge base of canine milk composition with the prospect of creating a quality, tailored milk substitute or supplement for puppies.
... An increase of the abundance of Bifidobacterium mongoliens is also observed in soft cheese during ripening. This last species could have an interest for human health, by the production of antivirulent metabolites (Bondue et al., 2020). ...
... In any case, the importance of these bacterial strains, capable of effectively metabolizing the oligosaccharides of breast milk, is due to the fact that they produce numerous essential substances for the human being, including some vitamins, protective factors against infections and short-chain fatty acids (SCFAs) [44,51,52]. Among these, the most important are: propionate, butyrate and acetate, which represent 90-95% of the SCFAs present in the colon [53]. ...
Human milk oligosaccharides (HMOs) are the third most represented component in breast milk. They serve not only as prebiotics but they exert a protective role against some significant neonatal pathologies such as necrotizing enterocolitis. Furthermore, they can program the immune system and consequently reduce allergies and autoimmune diseases’ incidence. HMOs also play a crucial role in brain development and in the gut barrier’s maturation. Moreover, the maternal genetic factors influencing different HMO patterns and their modulation by the interaction and the competition between active enzymes have been widely investigated in the literature, but there are few studies concerning the role of other factors such as maternal health, nutrition, and environmental influence. In this context, metabolomics, one of the newest “omics” sciences that provides a snapshot of the metabolites present in bio-fluids, such as breast milk, could be useful to investigate the HMO content in human milk. The authors performed a review, from 2012 to the beginning of 2021, concerning the application of metabolomics to investigate the HMOs, by using Pubmed, Researchgate and Scopus as source databases. Through this technology, it is possible to know in real-time whether a mother produces a specific oligosaccharide, keeping into consideration that there are other modifiable and unmodifiable factors that influence HMO production from a qualitative and a quantitative point of view. Although further studies are needed to provide clinical substantiation, in the future, thanks to metabolomics, this could be possible by using a dipstick and adding the eventual missing oligosaccharide to the breast milk or formula in order to give the best and the most personalized nutritional regimen for each newborn, adjusting to different necessities.
... α2-6-Linked sialylated oligosaccharides were present in greater proportion than the α2-3-linked structures during early lactation (167,168). Recently, Bondue et al. demonstrated the ability of a specific Bifidobacteria, Bifidobacterium mongoliense, to utilize 3 ′ -SL as the main source of carbon (169). ...
The assembly of the newborn's gut microbiota during the first months of life is an orchestrated process resulting in specialized microbial ecosystems in the different gut compartments. This process is highly dependent upon environmental factors, and many evidences suggest that early bacterial gut colonization has long-term consequences on host digestive and immune homeostasis but also metabolism and behavior. The early life period is therefore a “window of opportunity” to program health through microbiota modulation. However, the implementation of this promising strategy requires an in-depth understanding of the mechanisms governing gut microbiota assembly. Breastfeeding has been associated with a healthy microbiota in infants. Human milk is a complex food matrix, with numerous components that potentially influence the infant microbiota composition, either by enhancing specific bacteria growth or by limiting the growth of others. The objective of this review is to describe human milk composition and to discuss the established or purported roles of human milk components upon gut microbiota establishment. Finally, the impact of maternal diet on human milk composition is reviewed to assess how maternal diet could be a simple and efficient approach to shape the infant gut microbiota.
