Whole genome sequencing and characterization of Corynebacterium isolated from the healthy and dry eye ocular surface.

BACKGROUND: The purpose of this study was to characterize Corynebacterium isolated from the ocular surface of dry eye disease patients and healthy controls. We aimed to investigate the pathogenic potential of these isolates in relation to ocular surface health. To this end, we performed whole genome sequencing in combination with biochemical, enzymatic, and antibiotic susceptibility tests. In addition, we employed deferred growth inhibition assays to examine how Corynebacterium isolates may impact the growth of potentially competing microorganisms including the ocular pathogens Pseudomonas aeruginosa and Staphylococcus aureus, as well as other Corynebacterium present on the eye. RESULTS: The 23 isolates were found to belong to 8 different species of Corynebacterium with genomes ranging from 2.12 mega base pairs in a novel Corynebacterium sp. to 2.65 mega base pairs in C. bovis. Whole genome sequencing revealed the presence of a range of antimicrobial targets present in all isolates. Pangenome analysis showed the presence of 516 core genes and that the pangenome is open. Phenotypic characterization showed variously urease, lipase, mucinase, protease and DNase activity in some isolates. Attention was particularly drawn to a potentially new or novel Corynebacterium species which had the smallest genome, and which produced a range of hydrolytic enzymes. Strikingly the isolate inhibited in vitro the growth of a range of possible pathogenic bacteria as well as other Corynebacterium isolates. The majority of Corynebacterium species included in this study did not seem to possess canonical pathogenic activity. CONCLUSIONS: This study is the first reported genomic and biochemical characterization of ocular Corynebacterium. A number of potential virulence factors were identified which may have direct relevance for ocular health and contribute to the finding of our previous report on the ocular microbiome, where it was shown that DNA libraries were often dominated by members of this genus. Particularly interesting in this regard was the observation that some Corynebacterium, particularly new or novel Corynebacterium sp. can inhibit the growth of other ocular Corynebacterium as well as known pathogens of the eye.

Genomics of Staphylococcus aureus and Enterococcus faecalis isolated from the ocular surface of dry eye disease sufferers.

Ocular surface inflammatory disorders, such as dry eye, are becoming increasingly prevalent. Developing new treatment strategies targeting harmful bacteria could provide significant therapeutic benefits. The purpose of this study was to characterize the common ocular pathogen Staphylococcus aureus and the rarer endophthalmitis-associated species Enterococcus faecalis isolated from the ocular surface of dry eye disease patients in Norway. Together the 7 isolates (5 S. aureus and 2 E. faecalis) comprise the complete set of members of each species isolated in our previous study of the ocular microbiome of 61 dry eye sufferers. We aimed to investigate the pathogenic potential of these isolates in relation to ocular surface health. To this end, we used whole genome sequencing, multiplex PCR directed at virulence genes and antibiotic susceptibility tests encompassing clinically relevant agents. The E. faecalis isolates showed resistance to only gentamicin. S. aureus isolates displayed susceptibility to most of the tested antibiotics, except for two isolates which showed resistance to trimethoprim/sulfamethoxazole and three isolates which were resistant to ampicillin. Susceptibilities included sensitivity to several first-line antibiotics for treatment of ocular infections by these species. Thus, treatment options would be available if required. However, spontaneous resistance development to gentamicin and rifampicin occurred in some S. aureus which could be a cause for concern. Whole genome sequencing of the isolates showed genome sizes ranging from 2.74 to 2.83 Mbp for S. aureus and 2.86 Mbp for E. faecalis, which is typical for these species. Multilocus sequence typing and phylogenetic comparisons with previously published genomes, did not suggest the presence of eye-specific clusters for either species. Genomic analysis indicated a high probability of pathogenicity among all isolates included in the study. Resistome analysis revealed the presence of the beta-lactamase blaZ gene in all S. aureus isolates and the dfrG gene in two of them; while E. faecalis isolates carried the lsa(A) gene which confers intrinsic resistance to lincosamides and streptogramin A in this species. Screening for virulence factors revealed the presence of various pathogenicity associated genes in both S. aureus and E. faecalis isolates. These included genes coding for toxin production and factors associated with evading the host immune system. Some of the identified genes (tst, hylA & hylB) are suggested to be linked to the pathophysiology of dry eye disease. Lastly, the presence of specific S. aureus virulence genes was confirmed through multiplex PCR analysis.

Novel Teixobactin Analogues Show Promising In Vitro Activity on Biofilm Formation by Staphylococcus aureus and Enterococcus faecalis.

The treatment of infections caused by biofilm-forming organisms is challenging. The newly discovered antibiotic teixobactin shows activity against a wide range of biofilm-forming bacteria. However, the laborious and low-yield chemical synthesis of teixobactin complicates its further development for clinical application. The use of more easily synthesized teixobactin analogues may offer promise in this regard. In this article, three newly developed analogues were tested for efficacy against Staphylococcus aureus and Enterococcus faecalis. Minimum inhibitory and -bactericidal concentrations were investigated. MIC values for S. aureus and E. faecalis ranged from 0.5-2 and 2-4 µg/mL, respectively. Moreover, the ability of the analogues to prevent biofilm formation and to inactivate bacterial cells in already established S. aureus biofilm on medical grade materials (PVC and PTFE) used in the production of infusion tubing and catheters were also tested. The analogues showed an ability to prevent biofilm formation and inactivate bacterial cells in established biofilms at concentrations as low as 1-2 µg/mL. Confocal laser scanning microscopy showed that the most promising analogue (TB3) inactivated S. aureus cells in a preformed biofilm and gave a reduction in biovolume. The relative ease of synthesis of the analogues and their in vitro efficacy, makes them promising candidates for pharmaceutical development.

Positioning of community pharmacists in interactions with general practitioners and patients regarding prescribing and using antibiotics.

Interprofessional collaboration between general practitioners (GPs) and community pharmacists (CPs) is important for ensuring antibiotics are used correctly and combating antibiotic resistance. The study's main objective was to investigate how CPs, GPs and patients, respectively, position CPs in their interactions with patients on antibiotic-related matters in Norwegian pharmacies. Seven focus-group interviews were performed. Data were analyzed using systematic text condensation. Positioning theory was used to identify positions assigned to CPs by themselves, by GPs and by patients. CPs position themselves as helpful, accessible drug specialists responsible for advising on antibiotic use, but also consider themselves dependent on GP-supplied information to do so. GPs position CPs as helpful, responsible businesspeople who, however, lack clinical experience and are overzealous gatekeepers. Patients position CPs as helpful people who supply information in "everyday language" and as the GP's extended arm. Patients utter they are best served when GPs and CPs collaborate. This discrepancy is a barrier to optimal service to patients in general, and to proper antibiotic use in particular.

Diversion and phylogenetic relatedness of filterable bacteria from Norwegian tap and bottled waters.

Numerous articles have documented the existence of filterable bacteria. Where filtration is the chosen method of sterilization for medicinal or media components, these bacteria will by definition render products non-sterile. They may further represent a health hazard to the end user. A wide-range of bacterial genera were found in bottled and tap water filtrates from 0.2 µm filters, including genera housing opportunistic pathogens (e.g. Methylobacterium) and endospore formers (Paenibacillus). Two municipal tap water isolates were only distantly related to named species. One of these grew on agar, and could potentially provide hitherto unharvested useful biological products. The other grew only in water, and failed to produce colonies on media targeting either heterotrophs or autotrophs. The present study is one of very few looking at filterable bacteria in bottled waters intended for human consumption and the first identifying the filterable portion. It extends the range of known habitats of filterable bacteria and provides data on two new or novel species.

Implications of stx loss for clinical diagnostics of Shiga toxin-producing Escherichia coli.

The dynamics related to the loss of stx genes from Shiga toxin-producing Escherichia coli remain unclear. Current diagnostic procedures have shortcomings in the detection and identification of STEC. This is partly owing to the fact that stx genes may be lost during an infection or in the laboratory. The aim of the present study was to provide new insight into in vivo and in vitro stx loss in order to improve diagnostic procedures. Results from the study support the theory that loss of stx is a strain-related phenomenon and not induced by patient factors. It was observed that one strain could lose stx both in vivo and in vitro. Whole genome comparison of stx-positive and stx-negative isolates from the same patient revealed that different genomic rearrangements, such as complete or partial loss of the parent prophage, may be factors in the loss of stx. Of diagnostic interest, it was shown that patients can be co-infected with different E. coli pathotypes. Therefore, identification of eae-positive, but stx-negative isolates should not be interpreted as "Shiga toxin-lost" E. coli without further testing. Growth and recovery of STEC were supported by different selective agar media for different strains, arguing for inclusion of several media in STEC diagnostics.

Use of a Commercially Available Microarray to Characterize Antibiotic-Resistant Clinical Isolates of Klebsiella pneumoniae.

A commercially available microarray (IDENTIBAC AMR-ve) for the detection of antibiotic resistance determinants was investigated for its potential to genotype 30 clinical isolates and two control strains of Klebsiella pneumoniae. Resistance profiles and the production of extended-spectrum ß-lactamases were determined by disc diffusion and the results were compared with the microarray profiles in order to assess its scope and limitations. Genes associated with resistance to a wide range of antibiotics, including current first line therapy options, were detected. In addition, the array also detected class 1 integrases. The array is easy to use and interpret, and is useful in providing a general description of the numbers and types of resistance determinants in K. pneumoniae. It also provides an indication of the potential for resistance gene acquisition. However, in most instances detected resistance to specific antibiotics could not unequivocally be assigned to hybridization with a specific array probe. We conclude that the microarray is a valuable and rapid means of investigating the presence of resistance gene classes of therapeutic importance. It can also provide a starting point for selecting analyses of greater resolving power, such as phylogenetic subtyping by PCR sequencing.

Toxin production and antibiotic resistances in Escherichia coli isolated from bathing areas along the coastline of the Oslo fjord.

The presence of enterovirulent and/or antibiotic resistant strains of Escherichia coli in recreational bathing waters would represent a clear health issue. In total, 144 E. coli isolated from 26 beaches along the inner Oslo fjord were examined for virulence determinants and resistance to clinically important antibiotics. No isolates possessed the genetic determinants associated with enterotoxigenic strains and none showed the prototypic sorbitol negative, O157:H7 phenotype. A small number (∼1 %) produced alpha-hemolysin. Occurrences and patterns of antibiotic resistances were similar to those of E. coli isolated previously from environmental samples. In total, 6 % of the strains showed one or more clinically relevant resistances and 1.4 % were multi-drug resistant. Microarray analyses suggested that the resistance determinants were generally associated with mobile genetic elements. Resistant strains were not clonally related, and were, furthermore not concentrated at one or a few beach sites. This suggests that these strains are entering the waters at a low rate but in a widespread manner. The study demonstrates that resistant E. coli are present in coastal bathing waters where they can come into contact with bathers, and that the resistance determinants are potentially transferable. Some of the resistances registered in the study are to important antibiotics used in human medicine such as fluoroquinolones. The spread of antibiotic resistant genes, from the clinical setting to the environment, has clear implications with respect to the current management of bacterial infections and the long term value of antimicrobial therapy. The present study is the first of its kind in Norway.

Culture- and non-culture-based approaches reveal unique features of the ocular microbiome in dry eye patients.

PURPOSE: The purpose of this study was to investigate the ocular microbiome in individuals with dry eye disease and to identify features of their ocular microbiome of possible health and diagnostic significance. METHODS: Conjunctival samples were collected from both eyes in duplicate from 91 individuals (61 dry eye, 30 healthy) and used for both culture-dependent and culture-independent analyses. Samples were either analysed using next generation sequencing (V3-V4 16S rDNA) or inoculated on a wide range of agar types and grown under a broad range of conditions to maximize recovery. Isolates were identified by partial sequencing of the 16S rDNA and rpoB genes and tested for antibiotic susceptibility. We applied a L2-regularized logistic regression model on the next generation sequencing data to investigate any potential association between severe dry eye disease and the ocular microbiome. RESULTS: Culture-dependent analysis showed the highest number of colony forming units in healthy individuals. The majority of isolates recovered from the samples were Corynebacterium, Micrococcus sp., Staphylococcus epidermidis, and Cutibacterium acnes. Culture independent analysis revealed 24 phyla, of which Actinobacteria, Firmicutes and Proteobacteria were the most abundant. Over 405 genera were detected of which Corynebacterium was the most dominant, followed by Staphylococcus and Cutibacterium. The L2-regularized logistic regression model indicated that Blautia and Corynebacterium sp. may be associated with severe DED. CONCLUSIONS: Our study indicates that the ocular microbiome has characteristic features in severe DED patients. Certain Corynebacterium species and Blautia are of particular interest for future studies.

A preliminary study of the use of MinION sequencing to specifically detect Shiga toxin-producing Escherichia coli in culture swipes containing multiple serovars of this species.

An important challenge relating to clinical diagnostics of the foodborne pathogen Shiga toxin-producing E. coli (STEC), is that PCR-detection of the shiga-toxin gene (stx) in DNA from stool samples can be accompanied by a failure to identify an STEC isolate in pure culture on agar. In this study, we have explored the use of MinION long-read sequencing of DNA from bacterial culture swipes to detect the presence of STEC, and bioinformatic tools to characterize the STEC virulence factors. The online workflow "What's in my pot" (WIMP) in the Epi2me cloud service, rapidly identified STEC also when it was present in culture swipes together with multiple other E. coli serovars, given sufficient abundance. These preliminary results provide useful information about the sensitivity of the method, which has potential to be used in clinical diagnostic of STEC, particularly in cases where a pure culture of the STEC isolate is not obtained due to the 'STEC lost Shiga toxin' phenomenon.

Exploring patients' adherence to antibiotics by understanding their health knowledge and relational communication in encounters with pharmacists and physicians.

Background: Antibiotics are drugs essential for the treatment of bacterial infections. Widespread and often improper use of antibiotics are driving the emergence of antimicrobial resistance (AMR) globally. A better understanding of the communicated and understood use of antibiotics as well as improved adherence to treatments are needed to meet this public health threat. Objectives: The aim of the study is to explore how knowledge of antibiotic use is collected and communicated between patients, physicians, and pharmacists, and how patients seek, understand and use available information on antibiotics in adherence to prescribed treatment. Methods: Seven focus group interviews were conducted with community pharmacists (three groups, eleven participants), physicians/general practitioners (two groups, thirteen participants), and patients (two groups, eight participants) in Norway. Four focus group interviews were conducted offline and three online. The interview data were analyzed using systematic text condensation in a four-step, descriptive and explorative thematic analysis. Results: Three main themes were developed about patients' adherence to antibiotics: 1. patients' knowledge about antibiotics and AMR; 2. sources of information about antibiotics/AMR; and 3. relational communication. Patient knowledge about both antibiotics and AMR was somewhat limited, and showed considerable variation. Patients relied on the internet, chat sites, printed information, and face-to-face meetings with health professionals for information. Relational communication between patients, physicians, and pharmacists was found to be important in reducing misunderstandings.Vulnerability, limited time, and lack of communication were barriers to receiving and understanding information during patient-physician encounters. Increased knowledge about antibiotics and AMR may result in better adherence to prescribed medications. Conclusions: Patients seek information about antibiotics and AMR in three arenas; digital platforms, printed material and face to face encounters. However, patients often misunderstand important facts relating to this issue. Relational communication between patients, physicians, and pharmacists was important to ensure adherence to treatment regimens. Pharmacists are encouraged to use open-ended questions and build upon the information obtained from the physician to provide patients with tailored advice and ensure proper adherence. Pharmacists' contribution is crucial in optimizing antibiotic use and combating AMR, as they are the final healthcare point of contact before treatment initiation.

Activity of 6-aryl-pyrrolo[2,3-d]pyrimidine-4-amines to Tetrahymena.

A series 6-aryl-pyrrolo[2,3-d]pyrimidine-4-amines (43 compounds), some of which are epidermal growth factor tyrosine kinase inhibitors, were tested for their protozoal toxicity using an environmental Tetrahymena strain as model organism. The protozoacidal activity of the analogues was found to be highly dependent on a 4-hydroxyl group at the 6-aryl ring, and a chiral 1-phenylethanamine substituent in position 4. Further, the potency was affected by the aromatic substitution pattern of the phenylethanamine: the unsubstituted, the meta-fluoro and the para-bromo substituted derivatives had the lowest minimum protozoacidal concentrations (8-16 µg/mL). Surprisingly, both enantiomers were found to have high potency suggesting that this compound class could have several modes of action. No correlation was found between the compounds protozoacidal activity and the in vitro epidermal growth factor receptor tyrosine kinase inhibitory potency. This suggests that the observed antimicrobial effects are related to other targets. Testing towards a panel of kinases indicated several alternative modes of action.

Synthesis and antimycobacterial activities of non-purine analogs of 6-aryl-9-benzylpurines: Imidazopyridines, pyrrolopyridines, benzimidazoles, and indoles.

6,9-Disubstituted purines and 7-deazapurines are known to be powerful inhibitors of Mycobacterium tuberculosis (Mtb) in vitro. Analogs modified in the six-membered ring (imidazopyridines, pyrrolopyridines, benzimidazoles, and indoles) were synthesized and evaluated as Mtb inhibitors. The targets were prepared by functionalization on the bicyclic heterocycle or from simple pyridines. The results reported herein, indicate that the purine N-1, but not N-3, is important for binding to the unknown target. The 3-deazapurines appears to be slightly more active compared to the parent purines and slightly less active than their 7-deazapurine isomers. Removal of both the purine N-3 and N-7 did not result in further enhanced antimycobacterial activity but the toxicity towards mammalian cells was increased. Both 3-deaza and 3,7-dideazapurines exhibited a modest activity against of the Mtb isolate in the state of non-replicating persistence.

Antimicrobial and antineoplastic activities of agelasine analogs modified in the purine 2-position.

Agelasines are 7,9-dialkylpurinium salts found in marine sponges (Agelas sp.), which display a variety of antimicrobial and cytotoxic effects. We have synthesized simplified agelasine analogs modified in the purine 2-position and examined their antimicrobial and anticancer activities. The compounds were screened against Staphylococcus aureus, Escherichia coli, Mycobacterium tuberculosis, Candida krusei, and Candida albicans, protozoa causing tropical diseases (Plasmodium falciparum, Leishmania infantum, Trypanosoma cruzi, and Trypanosoma brucei), a panel of human cancer cell lines (U-937 GTB, RPMI 8226/s, CEM/s, and ACHN) as well as VERO and/or MRC-5 cells. The results indicate that the introduction of a methyl group in the purine 2-position is beneficial for antimycobacterial and antiprotozoal activity, and that amino groups may enhance activity against several cancer cell lines.

Norwegian Soils and Waters Contain Mesophilic, Plastic-Degrading Bacteria.

Plastic pollution has become one of the most critical environmental issues, as rapidly increasing production, compounded by persistence of plastic wastes in the environment, are outpacing efforts to keep ecosystems plastic-free. A switch to plastics more amenable to microbial attack is one of several possible responses. Against this background, the current study describes the isolation, enumeration and polyphasic characterization of plastic-degrading bacteria present in Norwegian terrestrial and aquatic habits. It shows that these bacteria are present in relatively high numbers, and that plastic-degrading capabilities are found in several taxa, most especially Streptomyces. Some isolates wereable to degrade several plastics. Notably, a Rhodococcus sp. and a Streptomyces sp. degraded, respectively, four and six of the eight plastics investigated and a number of other polymers relevant for plastic blends. The paper also has a methodological aspect, presenting various approaches for assaying plastic-degrading properties and a PCR/sequencing-based approach for the identification of potential polyethylene terephthalate-degrading genes. A candidate gene was detected in several Streptomyces isolates. The study shows that Norwegian environments are a rich source of bacteria with the ability to degrade bioplastics possibly representing a natural remediation capacity, as well as a potential source of useful enzymes.

A simple and novel method for the production of polyethylene terephthalate containing agar plates for the growth and detection of bacteria able to hydrolyze this plastic.

Polyethylene terephthalate is used in the manufacture of many products. Microbes able to hydrolyze the plastic are known, and offer promise for practical waste management. Screening for hydrolysis usually involves unwieldy culturing in the presence of film. A rapid screening method based on culturing on agar plates is here described.

The effect of toll-like receptor ligands on energy metabolism and myokine expression and secretion in cultured human skeletal muscle cells.

Skeletal muscle plays an important role in glycaemic control and metabolic homeostasis, making it a tissue of interest with respect to type 2 diabetes mellitus. The aim of the present study was to determine if ligands of Toll-like receptors (TLRs) could have an impact on energy metabolism and myokine expression and secretion in cultured human skeletal muscle cells. The myotubes expressed mRNA for TLRs 1-6. TLR3, TLR4, TLR5 and TLR6 ligands (TLRLs) increased glucose metabolism. Furthermore, TLR4L and TLR5L increased oleic acid metabolism. The metabolic effects of TLRLs were not evident until after at least 24 h pre-incubation of the cells and here the metabolic effects were more evident for the metabolism of glucose than oleic acid, with a shift towards effects on oleic acid metabolism after chronic exposure (168 h). However, the stimulatory effect of TLRLs on myokine expression and secretion was detected after only 6 h, where TLR3-6L stimulated secretion of interleukin-6 (IL-6). TLR5L also increased secretion of interleukin-8 (IL-8), while TLR6L also increased secretion of granulocyte-macrophage colony stimulating factor (GM-CSF). Pre-incubation of the myotubes with IL-6 for 24 h increased oleic acid oxidation but had no effect on glucose metabolism. Thus IL-6 did not mimic all the metabolic effects of the TLRLs, implying metabolic effects beyond the actions of this myokine.

Synthesis of non-purine analogs of 6-aryl-9-benzylpurines, and their antimycobacterial activities. Compounds modified in the imidazole ring.

Purine analogs modified in the five-membered ring have been synthesized and examined for antibacterial activity against Mycobacterium tuberculosis H(37)Rv in vitro employing the microplate alamar blue assay (MABA). The 9-deaza analogs were only found to be weak inhibitors, but the 8-aza-, 7-deaza- and 8-aza-7-deazapurine analogs studied displayed excellent antimycobacterial activities, some even substantially better than the parent purine. In the 7-deazapurine series, MIC values between 0.08 and 0.35 µM, values comparable or better than the reference drugs used in the study (MIC rifampicin 0.09 µM, MIC isoniazid 0.28 µM and MIC PA-824 0.44 µM). The five most active compounds were also examined against a panel of drug-resistant Mtb strain, and they all retained their activity. The compounds examined were significantly less active against M. tuberculosis in a state of non-replicating persistence (NRP). MIC in the low-oxygen-recovery assay (LORA) ≥ 60 µM. The 7-deazapurines were somewhat more toxic towards mammalian cells, but still the selectivity indexes were excellent. The non-purine analogs exhibit a selective antimycobacterial activity. They were essentially inactive against Staphylococcus aureus and Escherichia coli.

A preliminary study on the potential of Nanopore MinION and Illumina MiSeq 16S rRNA gene sequencing to characterize building-dust microbiomes.

There is a growing awareness of the importance of indoor microbiomes for human health. Given their complexity, these microbiomes can only be adequately surveyed using high throughput sequencing techniques. Oxford Nanopore's MinION is the newest third generation sequencing technology on the market. With its many advantages such as portability, user friendliness, simplicity, speed of sequencing and long read length, the technology is now an actual contender to established sequencing platforms. MinION's main disadvantage is a relatively low read accuracy compared to several other platforms, although this is constantly improving. The present study, which appears to be the first of its kind, provides the results of a preliminary analysis of the microbial communities in indoor environments based on 16S rRNA gene amplicon sequencing, using both the Oxford Nanopore Technologies (ONT) MinIOn and the Illumina MiSeq DNA sequencers. At the level of family and above, there was no significant difference between the microbial compositions as revealed by the two platforms. However, at the genus, and particularly at the species level, the ONT MinION reported greater taxonomic resolution than Illumina MiSeq.

Dietary Fiber, Gut Microbiota, and Metabolic Regulation-Current Status in Human Randomized Trials.

New knowledge about the gut microbiota and its interaction with the host's metabolic regulation has emerged during the last few decades. Several factors may affect the composition of the gut microbiota, including dietary fiber. Dietary fiber is not hydrolyzed by human digestive enzymes, but it is acted upon by gut microbes, and metabolites like short-chain fatty acids are produced. The short-chain fatty acids may be absorbed into the circulation and affect metabolic regulation in the host or be a substrate for other microbes. Some studies have shown improved insulin sensitivity, weight regulation, and reduced inflammation with increases in gut-derived short-chain fatty acids, all of which may reduce the risk of developing metabolic diseases. To what extent a dietary intervention with fiber may affect the human gut microbiota and hence metabolic regulation, is however, currently not well described. The aim of the present review is to summarize recent research on human randomized, controlled intervention studies investigating the effect of dietary fiber on gut microbiota and metabolic regulation. Metabolic regulation is discussed with respect to markers relating to glycemic regulation and lipid metabolism. Taken together, the papers on which the current review is based, suggest that dietary fiber has the potential to change the gut microbiota and alter metabolic regulation. However, due to the heterogeneity of the studies, a firm conclusion describing the causal relationship between gut microbiota and metabolic regulation remains elusive.