Microbial Research Articles

The effect of in vitro simulated colonic pH gradients on microbial activity and metabolite production using common prebiotics as substrates

BackgroundThe interplay between gut microbiota (GM) and the metabolization of dietary components leading to the production of short-chain fatty acids (SCFAs) is affected by a range of factors including colonic pH and carbohydrate source. However, there is still only limited knowledge on how the GM activity and metabolite production in the gastrointestinal tract could be influenced by pH and the pH gradient increases along the colon.ResultsHere we investigate the effect of pH gradients corresponding to levels typically found in the colon on GM composition and metabolite production using substrates inulin, lactose, galactooligosaccharides (GOS), and fructooligosaccharide (FOS) in an in vitro colon setup. We investigated 3 different pH regimes (low, 5.2 increasing to 6.4; medium, 5.6 increasing to 6.8 and high, 6.0 increasing to 7.2) for each fecal inoculum and found that colonic pH gradients significantly influenced in vitro simulated GM structure, but the influence of fecal donor and substrate was more pronounced. Low pH regimes strongly influenced GM with the decreased relative abundance of Bacteroides spp. and increased Bifidobacterium spp. Higher in vitro simulated colonic pH promoted the production of SCFAs in a donor- and substrate-dependent manner. The butyrate producer Butyricimonas was enriched at higher pH conditions, where also butyrate production was increased for inulin. The relative abundance of Phascolarctobacterium, Bacteroides, and Rikenellaceae also increased at higher colonic pH, which was accompanied by increased production of propionate with GOS and FOS as substrates.ConclusionsTogether, our results show that colonic substrates such as dietary fibres influence GM composition and metabolite production, not only by being selectively utilized by specific microbes, but also because of their SCFA production, which in turn also influences colonic pH and overall GM composition and activity. Our work provides details about the effect of the gradients of rising pH from the proximal to distal colon on fermenting dietary substrates in vitro and highlights the importance of considering pH in GM research.

Passivating the Background of Living Microbes with a Zwitterionic Peptide for Therapies.

Living microbial therapies have been proposed as a course of action for a variety of diseases. However, problematic interactions between the host immune system and the microbial organism present significant clinical concerns. Previously, we developed a genetically encoded superhydrophilic zwitterionic peptide, termed EKP, to mimic low-immunogenic zwitterionic materials, which have been used for the chemical modification of biologics such as protein and nucleic acid drugs to increase their in vivo circulation time and reduce their immunogenicity. Herein, we demonstrate the protective effects of the EKP polypeptide genetically cloaking the surface of Saccharomyces cerevisiae as a model microbe in both in vitro and in vivo systems. First, we show that EKP peptide cloaking suppresses the interactions between yeast cells and their specific antibodies, thereby illustrating its cloaking behavior. Then, we examine the in vitro interactions between EKP peptide surface cloaked yeast cells and murine macrophage cells, which exhibit phagocytotic behavior in the presence of foreign microbes. Our results indicate that EKP cloaking suppresses macrophage interactions and thus reduces phagocytosis. Furthermore, EKP cloaked yeast cells demonstrate a prolonged circulation time in mice in vivo.

Current trends in medium‐chain‐length polyhydroxyalkanoates: Microbial production, purification, and characterization

AbstractPolyhydroxyalkanoates (PHAs) have gained interest recently due to their biodegradability and versatility. In particular, the chemical compositions of medium‐chain‐length (mcl)‐PHAs are highly diverse, comprising different monomers containing 6–14 carbon atoms. This review summarizes different feedstocks and fermentation strategies to enhance mcl‐PHA production and briefly discusses the downstream processing. This review also provides comprehensive details on analytical tools for determining the composition and properties of mcl‐PHA. Moreover, this study provides novel information by statistically analyzing the data collected from several reports on mcl‐PHA to determine the optimal fermentation parameters (specific growth rate, PHA productivity, and PHA yield from various structurally related and unrelated substrates), mcl‐PHA composition, molecular weight (MW), and thermal and mechanical properties, in addition to other relevant statistical values. The analysis revealed that the median PHA productivity observed in the fed‐batch feeding strategy was 0.4 g L−1 h−1, which is eight times higher than that obtained from batch feeding (0.05 g L−1 h−1). Furthermore, 3‐hydroxyoctanoate and ‐decanoate were the primary monomers incorporated into mcl‐PHA. The investigation also determined the median glass transition temperature (−43°C) and melting temperature (47°C), which indicated that mcl‐PHA is a flexible amorphous polymer at room temperature with a median MW of 104 kDa. However, information on the monomer composition or heterogeneity and the associated physical and mechanical data of mcl‐PHAs is inadequate. Based on their mechanical values, the mcl‐PHAs can be classified as semi‐crystalline polymers (median crystallinity 23%) with rubber‐like properties and a median elongation at break of 385%. However, due to the limited mechanical data available for mcl‐PHAs with known monomer composition, identifying suitable processing tools and applications to develop mcl‐PHAs further is challenging.

Production of a Rich Fertilizer Base for Plants from Waste Organic Residues by Microbial Formulation Technology.

This review explores different methods of sustainably introducing nutrients from agro-industrial waste into the soil. The focus is on sustainable agriculture and how the soil system can be modified by introducing secondary raw materials and beneficial microorganisms. Soil is a nexus between plants and microorganisms that must be balanced. The article emphasizes the importance of maintaining the microbiological balance when supplying nutrients. This review is focused on the possible techniques involved in the production of biofertilizers and their mode of application into the soil system and on plants. We addressed several advantages concerning the use of beneficial microorganisms in waste management by microbial formulation techniques. Not only the advantages but several limitations and challenges were also discussed in regard to the large scale production of microbial products. Therefore, the proper treatment of industrial waste is essential so that we can preserve the environment and human safety and also achieve sustainable agriculture.

Isolation and characterization of multiple-stress tolerant bacteria from radon springs.

Radon springs, characterized by their high concentrations of radon gas (Rn222), are extreme environments with unique physicochemical conditions distinct from conventional aquatic ecosystems. Our research aimed to investigate microbial life in radon springs, focusing on isolating extremophilic bacteria and assessing their resistance to adverse conditions. Our study revealed the prevalence of Actinomycetia species in the radon spring environment. We conducted various tests to evaluate the resistance of these isolates to oxidative stress, irradiation, desiccation, and metal ion content. These extremophilic bacteria showed overall higher resistance to these stresses compared to control strains. Lipidomic analysis was also employed to provide insights into the adaptive mechanisms of these bacteria which were found mainly in the correlations among individual clusters and changes in content of fatty acids (FA) as well as differences between content and type of FAs of environmental isolates and type strains.

Investigating the spatiotemporal dynamics of apple tree phyllosphere bacterial and fungal communities across cultivars in orchards.

The phyllosphere, a reservoir of diverse microbial life associated with plant health, harbors microbial communities that are subject to various complex ecological processes acting at multiple scales. In this study, we investigated the determinants of the spatiotemporal variation in bacterial and fungal communities within the apple tree phyllosphere, employing 16S and ITS amplicon sequencing. Our research assessed the impact of key factors - plant compartment, site, time, and cultivar - on the composition and diversity of leaf and flower microbial communities. Our analyses, based on samples collected from three cultivars in three orchards in 2022, revealed that site and time are the strongest drivers of apple tree phyllosphere microbial communities. Conversely, plant compartment and cultivar exhibited minor roles in explaining community composition and diversity. Predominantly, bacterial communities comprised Hymenobacter (25%) and Sphingomonas (10%), while the most relatively abundant fungal genera included Aureobasidium (27%) and Sporobolomyces (10%). Additionally, our results show a gradual decrease in alpha-diversity throughout the growth season. These findings emphasize the necessity to consider local microbial ecology dynamics in orchards, especially as many groups worldwide aim for the development of biocontrol strategies (e.g., by manipulating plant-microbe interactions). More research is needed to improve our understanding of the determinants of time and site-specific disparities within apple tree phyllosphere microbial communities across multiple years, locations, and cultivars.

Valorisation of Sugarcane Bagasse for the Sustainable Production of Polyhydroxyalkanoates

With the world shifting towards renewable and sustainable resources, polyhydroxyalkanoates (PHAs) have attracted significant interest as an alternative to synthetic plastics. While possessing promising properties suitable for various applications, the production of PHAs has not yet reached a global commercial scale. The main reason is the high cost of production, which represents a major limitation. Sugarcane bagasse (SCB) is an abundant lignocellulosic waste around the world. Its use to produce PHA enhances the feasibility of producing PHAs at commercial scale. However, SCB requires pretreatment and hydrolysis steps to release the sugars prior to the microbial fermentation. The cost associated with these steps poses additional challenges for large-scale production. Another challenge is the release of inhibitors during the pretreatment process which can result in a low PHA yield. The development of a low cost, co-culture strategy for the bioconversion of SCB into PHAs, can represent a pivotal step towards the large-scale production of bioplastics. This review highlights the advancements made in recent years on the microbial production of PHA using SCB as potential feedstock, with a proposed biological strategy and circular economy model.

Effects of pectin methyl-esterification on intestinal microbiota and its immunomodulatory properties in naive mice

Pectins are dietary fibers that are attributed with several beneficial immunomodulatory effects. Depending on the degree of esterification (DE), pectins can be classified as high methoxyl pectin (HMP) or low methoxyl pectin (LMP).The aim of this study was to investigate the effects of pectin methyl-esterification on intestinal microbiota and its immunomodulatory properties in naive mice.Supplementation of the diet with LMP or HMP induced changes in the composition of the intestinal microbiota in mice toward Bacteroides, which was mainly promoted by HMP. Metabolome analysis of stool samples from pectin-fed mice showed a different effect of the two types of pectin on the levels of short-chain fatty acids and bile acids, which was consistent with highly efficient in vivo fermentation of LMP. Analysis of serum antibody levels showed a significant increase in IgG and IgA levels by both pectins, while FACS analysis revealed a decrease of infiltrating inflammatory cells in the intestinal lamina propria by HMP.Our study revealed that the structural properties of the investigated pectins determine fermentability, effects on microbial composition, metabolite production, and modulation of immune responses. Consumption of HMP preferentially altered the gut microbiota and suppressed pro-inflammatory immune responses, suggesting a beneficial role in inflammatory diseases.

Sequential recovery of extracellular alginate and intracellular polyhydroxyalkanoate (PHA) from mixed microbial culture PHA production system

Polyhydroxyalkanoate (PHA) is a biodegradable plastic that exhibits properties similar to petroleum-based plastics. The mixed microbial culture (MMC) process utilized a waste carbon source to produce PHA, reducing the raw material costs associated with pure bacterial technology. However, the high cost of PHA recovery and environmental pollution are significant challenges to the sustainable development of MMC technology. In this study, a sequential recovery strategy was proposed. This strategy involved the use of Na2CO3 to break down the structural extracellular polymeric substances and rhamnolipid to digest non-PHA cell material (NPCM). The recovery rate of PHA in this strategy achieved 61.29 ± 5.92% and a purity of 97.55 ± 0.92%. Additionally, the strategy allowed the sequential recovery of the by-product alginate-like extracellular polymers. FT-IR analysis confirmed that the recovered biopolymer was PHA material, possessing mechanical and thermal properties similar to those obtained through CHCl3 extraction. However, it was important to note that this method had the potential to alter the distribution structure of the PHA polymer, although it successfully avoids the issue of reducing the molecular weight of PHA during NPCM digestion. A comprehensive evaluation demonstrates that the cost of PHA production using this method was only 14.61 CNY/kg, with a more positive environmental impact in terms of global warming potential and carbon emission reduction. Therefore, the sequential recovery strategy presents a feasible alternative for the downstream recovery process and overall economy of the MMC PHA process.

Concentration-dependent dual roles of proanthocyanidins on oxidative stress and docosahexaenoic acid production in Schizochytrium sp. ATCC 20888

Antioxidant addition is an effective strategy to achieve docosahexaenoic acid (DHA) overproduction in oleaginous microorganisms. Nevertheless, antioxidants like phenolic compounds sometimes exert pro-oxidant activity. In this work, effects of proanthocyanidins (PAs) on fermentation performance and oxidative stress in Schizochytrium sp. were investigated. Low PAs addition (5 mg/L) reduced reactive oxygen species and enhanced lipogenic enzymes activities and NADPH, resulting in significant increase in lipid (20.3 g/L) by 33.6 % and DHA yield (9.8 g/L) by 53.4 %. In contrast, high PAs addition (500 mg/L) exerted pro-oxidant effects, aggravated oxidative damage and lipid peroxidation, leading to sharp decrease in biomass (21.3 g/L) by 35.1 %, lipid (8.2 g/L) by 46.0 %, and DHA (2.9 g/L) by 54.8 %. Therefore, the antioxidant concentration is especially crucial in DHA production. This study is the first to report concentration-dependant dual roles of PAs in oxidative stress and DHA production in Schizochytrium sp., providing new insights into microbial DHA production.

Systematic Engineering of Genistein Biosynthetic Pathway through Genetic Regulators and Combinatorial Enzyme Screening.

Microbial production of genistein, an isoflavonoid primarily found in soybeans, is gaining prominence in the food industry due to its significant nutritional and health benefits. However, challenges arise in redesigning strains due to intricate regulatory nodes between cell growth and genistein production and in systematically exploring core enzymes involving genistein biosynthesis. To address this, this study devised a strategy that simultaneously and precisely rewires flux at both acetyl-CoA and malonyl-CoA nodes toward genistein synthesis. In particular, naringenin, the primary precursor of genistein, was accumulated 2.6 times more than the unoptimized strain through transcriptional repressor-based genetic regulators. Building upon this, a combination of isoflavone synthase and cytochrome P450 reductase with the remarkable conversion of naringenin to genistein was screened from enzyme homologue libraries. The integrated metabolic engineering strategy yields the highest reported production (98 mg/L of genistein) to date, providing a framework for the biosynthesis of diverse flavonoids, including genistein.

PEMANFAATAN LIMBAH SERBUK GERGAJIAN KAYU ULIN (Eusideroxylon zwageri T & B) UNTUK MENURUNKAN KADAR FECAL COLIFORM AIR SUNGAI

Martapura river water is included in categories that have a fairly high bacterial content, which is 21,333 colonies/100 ml. The optimal value of water quality standards to be met with water treatment businesses is the use of ironwood sawdust waste. Bacteria in water attach most of their lives to suspension particles, so microscopic organisms are also separated or filtered. The purpose of this study was to analyze fecal coliform in river water and analyze the volume of ironwood sawdust to reduce fecal coliform levels in river water. Martapura river water contains the highest fecal 1,800MPN/100ml and the lowest 1,600MPN/100 ml, according to the Minister of the Republic of Indonesia No. 22 of 2021 concerning the implementation of environmental protection and management related to standards, including category two which is used as water tourism facilities/infrastructure, freshwater fish farming, animal husbandry, plant irrigation and other activities requiring the same water quality and use. Ironwood sawdust as much as 25% or 90 grams proved most effective to be used to reduce fecal levels with an average value on the first day of 1,650 MPN/100ml and on the third day with treatment carried out up to indigo 350 MPN/100ml, so that the difference reached a value of 1,300 MPN/100ml.

Maternal smoking during pregnancy increases the risk of gut microbiome-associated childhood overweight and obesity

ABSTRACT Childhood obesity is linked to maternal smoking during pregnancy. Gut microbiota may partially mediate this association and could be potential targets for intervention; however, its role is understudied. We included 1,592 infants from the Canadian Healthy Infants Longitudinal Development Cohort. Data on environmental exposure and lifestyle factors were collected prenatally and throughout the first three years. Weight outcomes were measured at one and three years of age. Stool samples collected at 3 and 12 months were analyzed by sequencing the V4 region of 16S rRNA to profile microbial compositions and magnetic resonance spectroscopy to quantify the metabolites. We showed that quitting smoking during pregnancy did not lower the risk of offspring being overweight. However, exclusive breastfeeding until the third month of age may alleviate these risks. We also reported that maternal smoking during pregnancy significantly increased Firmicutes abundance and diversity. We further revealed that Firmicutes diversity mediates the elevated risk of childhood overweight and obesity linked to maternal prenatal smoking. This effect possibly occurs through excessive microbial butyrate production. These findings add to the evidence that women should quit smoking before their pregnancies to prevent microbiome-mediated childhood overweight and obesity risk, and indicate the potential obesogenic role of excessive butyrate production in early life.

Microbial production of dextran using pineapple waste extract: a two-step statistical optimization of submerged fermentation conditions and structural characterization

Microbial production of dextran using pineapple waste extract: a two-step statistical optimization of submerged fermentation conditions and structural characterization

Activity-Based Protein Profiling Identifies Protein Disulfide-Isomerases as Target Proteins of the Volatile Salinilactones.

The salinilactones, volatile marine natural products secreted from Salinispora arenicola, feature a unique [3.1.0]-lactone ring system and cytotoxic activities through a hitherto unknown mechanism. To find their molecular target, an activity-based protein profiling with a salinilactone-derived probe is applied that disclosed the protein disulfide-isomerases (PDIs) as the dominant mammalian targets of salinilactones, and thioredoxin (TRX1) as secondary target. The inhibition of protein disulfide-isomerase A1 (PDIA1) and TRX1 is confirmed by biochemical assays with recombinant proteins, showing that (1S,5R)-salinilactone B is more potent than its (1R,5S)-configured enantiomer. The salinilactones bound covalently to C53 and C397, the catalytically active cysteines of the isoform PDIA1 according to tandem mass spectrometry. Reactions with a model substrate demonstrated that the cyclopropyl group is opened by an attack of the thiol at C6. Fluorophore labeling experiments showed the cell permeability of a salinilactone-BODIPY (dipyrrometheneboron difluoride) conjugate and its co-localization with PDIs in the endoplasmic reticulum. The study is one of the first to pinpoint a molecular target for a volatile microbial natural product, and it demonstrates that salinilactones can achieve high selectivity despite their small size and intrinsic reactivity.

Microbiome abundance patterns as attractors and the implications for the inference of microbial interaction networks

Inferring microbial interaction networks from abundance patterns is an important approach to advance our understanding of microbial communities in general and the human microbiome in particular. Here we suggest discriminating two levels of information contained in microbial abundance data: (1) the quantitative abundance values and (2) the pattern of presences and absences of microbial organisms. The latter allows for a binary view on microbiome data and a novel interpretation of microbial data as attractors, or more precisely as fixed points, of a Boolean network.Starting from these attractors, our aim is to infer an interaction network between the species present in the microbiome samples. To accomplish this task, we introduce a novel inference method that combines the previously published ESABO (Entropy Shifts of Abundance vectors under Boolean Operations) method with an evolutionary algorithm. The key idea of our approach is that the inferred network should reproduce the original set of (observed) binary abundance patterns as attractors.We study the accuracy and runtime properties of this evolutionary method, as well as its behavior under incomplete knowledge of the attractor sets. Based on this theoretical understanding of the method we then show an application to empirical data.

Two-Phase Fermentation Systems for Microbial Production of Plant-Derived Terpenes.

Microbial cell factories, renowned for their economic and environmental benefits, have emerged as a key trend in academic and industrial areas, particularly in the fermentation of natural compounds. Among these, plant-derived terpenes stand out as a significant class of bioactive natural products. The large-scale production of such terpenes, exemplified by artemisinic acid-a crucial precursor to artemisinin-is now feasible through microbial cell factories. In the fermentation of terpenes, two-phase fermentation technology has been widely applied due to its unique advantages. It facilitates in situ product extraction or adsorption, effectively mitigating the detrimental impact of product accumulation on microbial cells, thereby significantly bolstering the efficiency of microbial production of plant-derived terpenes. This paper reviews the latest developments in two-phase fermentation system applications, focusing on microbial fermentation of plant-derived terpenes. It also discusses the mechanisms influencing microbial biosynthesis of terpenes. Moreover, we introduce some new two-phase fermentation techniques, currently unexplored in terpene fermentation, with the aim of providing more thoughts and explorations on the future applications of two-phase fermentation technology. Lastly, we discuss several challenges in the industrial application of two-phase fermentation systems, especially in downstream processing.

New trends in microbial lipid-based biorefinery for fermentative bioenergy production from lignocellulosic biomass

Using oleaginous microbial lipid-based biorefinery from lignocellulosic biomass (LCB) to produce fermentative bioenergy (i.e., biodiesel) represents an innovative second-generation fuel production technology. These lipids are predominantly intracellular triglycerides that accumulate through the metabolism of sugars in fermentation following pretreatment and enzymatic hydrolysis of LCB. This review investigates the recent advances in the microbial lipid production from LCB, focusing on the factors influencing the lead microbial lipid producers, different pretreatment methods (i.e., physical, chemical, biological, and combined pretreatment), enzymatic hydrolysis approaches, novel bioprocessing strategies (i.e., microbes-specific and fermentation model specific), and engineering techniques of the oleaginous microbes (i.e., genetic and metabolic alterations). The study demonstrates that oleaginous yeasts can synthesize significantly higher quantities of lipids when incorporated into the system, known as separated hydrolysis and lipid production, following various combined pretreatment methods. Interestingly, CRISPR is found to be the most suitable way of engineering microbes genetically and metabolically for increased lipid synthesis. The study also explores economically viable strategies for fermentative lipid production, addressing associated challenges, and outlines future directions, including comprehensive techno-economic and life cycle assessments. This review offers invaluable insights into microbial lipid production from LCB, highlighting the potential for significant technological and environmental enhancements through ongoing research and development efforts.

Microwave disinfection of facial silicone prostheses, Part 1: Color stability

The choice of cleaning method is an important consideration for lengthening the serviceable time of facial prostheses as microbial organisms and biofilms could degrade facial prostheses and cause skin irritation. Whether microwave disinfection is a suitable cleaning method without degradation of the properties of a prosthesis is unclear. The purpose of this in vitro study was to measure the color stability of 6 commonly used facial silicone elastomers after microwave disinfection over a simulated 1.5-year period. Six different facial silicone elastomers: MDX4-4210, MDX4-4210/Type A, M511, A-2186, VST-50, and A-2000 were mixed with intrinsic silicone white opacifier (except for the control group) and subsequently combined with 4 silicone intrinsic pigment color groups: red (R), yellow (Y), burnt sienna (B), and a mixture of R+Y+B (M). The control group was a silicone elastomer without opacifier or pigment. Each of the 30 experimental groups consisted of 5 specimens (N=150). Five specimens were placed in a 250-mL Erlenmeyer flask filled with 160mL of tap water. Seven flasks were then placed in a 660-W microwave oven. An exposure of 6 minutes was used according to the antimicrobial efficacy of microwave disinfection protocol on facial silicone prostheses with a final water temperature of 60°C for 18 times (simulating 1.5 years of microwave disinfection with one 6-minute exposure monthly). A spectrophotometer was used to measure reflectance color change values (∆E). Color differences were calculated following CIELab (∆E*ab) and CIEDE2000 (∆E00) formulae. ∆E*ab and ∆E00 were statistically analyzed by a linear mixed effects model with 3 factors (silicone type, color shade, and time) using the R Statistical software program (α=.05). Both ∆E*ab and ∆E00 of all silicone elastomers studied were less than the visual perceptibility thresholds (∆E*ab<1.1 and E00<0.7) and were considered clinically acceptable (∆E*ab<3.0 and E00<2.1) after the 1.5-year simulation of microwave disinfection. Yellow and blue pigments had more effect on MDX4-4210 and M511, while red pigment had more effect on MDX4-4210, MDX4-4210/Type A, and M511 (P<.05). Nevertheless, the values were still below the perceptibility threshold (∆E*ab≤1.0 and E00<0.6). All 6 facial silicone elastomers maintained clinically acceptable color after 18 months of exposure to microwave disinfection.

High fat diet is associated with gut microbiota dysbiosis and decreased gut microbial derived metabolites related to metabolic health in young Göttingen Minipigs.

The objectives were 1) to characterize a Göttingen Minipig model of metabolic syndrome regarding its colon microbiota and circulating microbial products, and 2) to assess whether ovariectomized female and castrated male minipigs show similar phenotypes. Twenty-four nine-week-old Göttingen Minipigs were allocated to four groups based on sex and diet: ovariectomized females and castrated males fed either chow or high-fat diet (HFD) for 12 weeks. At study end, body composition and plasma biomarkers were measured, and a mixed meal tolerance test (MMT) and an intravenous glucose tolerance test (IVGTT) were performed. The HFD groups had significantly higher weight gain, fat percentage, fasting plasma insulin and glucagon compared to the chow groups. Homeostatic model assessment of insulin resistance index (HOMA-IR) was increased and glucose effectiveness derived from the IVGTT and Matsuda´s insulin sensitivity index from the MMT were decreased in the HFD groups. The HFD groups displayed dyslipidemia, with significantly increased total-, LDL- and HDL-cholesterol, and decreased HDL/non-HDL cholesterol ratio. The colon microbiota of HFD minipigs clearly differed from the lean controls (GuniFrac distance matrix). The main bacteria families driving this separation were Clostridiaceae, Fibrobacteraceae, Flavobacteriaceae and Porphyromonadaceae. Moreover, the species richness was significantly decreased by HFD. In addition, HFD decreased the circulating level of short chain fatty acids and beneficial microbial metabolites hippuric acid, xanthine and trigonelline, while increasing the level of branched chain amino acids. Six and nine metabolically relevant genes were differentially expressed between chow-fed and HFD-fed animals in liver and omental adipose tissue, respectively. The HFD-fed pigs presented with metabolic syndrome, gut microbial dysbiosis and a marked decrease in healthy gut microbial products and thus displayed marked parallels to human obesity and insulin resistance. HFD-fed Göttingen Minipig therefore represents a relevant animal model for studying host-microbiota interactions. No significant differences between the castrated and ovariectomized minipigs were observed.