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MSystems Dec 2023This study sheds light on that treatment with but not is entitled to protect against necrotizing enterocolitis (NEC) development potentially. The mechanisms behind the...
This study sheds light on that treatment with but not is entitled to protect against necrotizing enterocolitis (NEC) development potentially. The mechanisms behind the opposite effect on NEC may result in different modulation on the level of , which is deeply associated with intestinal homoeostasis. Briefly, through improving the abundance of to alleviate intestinal inflammation and enhance intestinal barrier integrity, supplement may become a promising therapy for NEC.
Topics: Female; Infant, Newborn; Humans; Probiotics; Clostridium butyricum; Clostridium tyrobutyricum; Enterocolitis, Necrotizing; Intestines; Infant, Newborn, Diseases; Fetal Diseases
PubMed: 37921463
DOI: 10.1128/msystems.00732-23 -
Frontiers in Microbiology 2023Poor quality silage can derive from the presence of deleterious microorganisms such as clostridia. Their dissemination along the food chain, especially in milk, causes...
INTRODUCTION
Poor quality silage can derive from the presence of deleterious microorganisms such as clostridia. Their dissemination along the food chain, especially in milk, causes issues such as the cheese late-blowing defect, particularly triggered by . The scope of our study was to determine the occurrence in three different farms across four time periods in relation to the animal diets, specifically the Total Mixed Ration (TMR), by using real-time PCR.
METHODS
For this purpose, molecular-derived data were exploited to optimize a predictive model that simulated the farm conditions favoring the growth of butyric acid bacteria such as .
RESULTS
Our results showed that the originally utilized predictive model strongly underestimated the growth of in comparison to the molecular data. At the same time, our findings uncovered an additional source of contamination in the TMR related to silage and dietary residues that represent a reservoir of microbial contamination during successive TMR preparation. Based on these findings, the optimization of the model parameters such as growth rate range and the inclusion of the residues in the model, allowed a more accurate prediction of the contamination levels. Therefore, this study revealed that proper hygiene practices such as the removal of silage and TMR residues within the farm environment is essential to control the contamination by and avoid food waste and economic losses.
PubMed: 37051517
DOI: 10.3389/fmicb.2023.1118646 -
Nutrients Jan 2021This study aimed to investigate the effects of on colonic immunity and the role of IL-22 in the protective function of . Mice were supplemented with 10 CFU/mL daily...
This study aimed to investigate the effects of on colonic immunity and the role of IL-22 in the protective function of . Mice were supplemented with 10 CFU/mL daily for 20 days, followed by injecting with LPS for 24 h. In vivo interference of IL-22 via injecting with an adeno-associated virus was conducted to elucidate the role of IL-22 in attenuating colonic inflammation. The results showed that decreased the mRNA expression of IL-6 and IL-1β. enhanced the mRNA expression of IL-22 and the expression of MUC2 in the colon. The in vivo interference results showed that enhanced the mRNA expression of IL-6 and IL-1β while decreased the expression of MUC2 after knocking down IL-22. The flow cytometric analysis showed that decreased the proportions of macrophages, DCs, and mast cells and effectively regulated the proportion of Th17 cells, indicating that may stimulate the expression of IL-22 via regulating Th17 cells. Our study concluded that protected against LPS-induced colonic barrier dysfunction and inflammation via IL-22 signaling, suggesting that could be a potential probiotic in regulating colonic health.
Topics: Animals; Clostridium tyrobutyricum; Colitis; Dendritic Cells; Disease Models, Animal; Immunomodulation; Interleukins; Intestinal Mucosa; Lipopolysaccharides; Macrophages; Male; Mast Cells; Mice; Mice, Knockout; Probiotics; Signal Transduction; T-Lymphocyte Subsets; Interleukin-22
PubMed: 33451114
DOI: 10.3390/nu13010215 -
Microorganisms Nov 2020, a Gram-positive, anaerobic, spore-forming bacterium, is considered as one of the main causative agents for spoilage of hard and semihard cheeses. Growth of in cheese...
, a Gram-positive, anaerobic, spore-forming bacterium, is considered as one of the main causative agents for spoilage of hard and semihard cheeses. Growth of in cheese is critically influenced by ripening temperature and time, pH, salt and lactic acid concentration, moisture and fat content, and the presence of other microorganisms. Previous studies revealed high intraspecies diversity of strains and variable tolerance toward pH, temperatures, and salt concentrations. These findings indicate that strain-dependent characteristics may be relevant to assess the risk for cheese spoilage if clostridial contamination occurs. In this study, we aimed to compare the phenotypes of 12 strains which were selected from 157 strains on the basis of genotypic and proteotypic variability. The phenotypic analysis comprised the assessment of gas production and organic acid concentrations in an experimental cheese broth incubated at different temperatures (37, 20, and 14 °C). For all tested strains, delayed gas production at lower incubation temperatures and a strong correlation between gas production and the change in organic acid concentrations were observed. However, considering the time until gas production was visible at different incubation temperatures, a high degree of heterogeneity was found among the tested strains. In addition, variation among replicates of the same strain and differences due to different inoculum levels became evident. This study shows, that, among other factors, strain-specific germination and growth characteristics should be considered to evaluate the risk of cheese spoilage by .
PubMed: 33266400
DOI: 10.3390/microorganisms8111836 -
MBio Jun 2016Clostridium tyrobutyricum is a Gram-positive anaerobic bacterium that efficiently produces butyric acid and is considered a promising host for anaerobic production of...
UNLABELLED
Clostridium tyrobutyricum is a Gram-positive anaerobic bacterium that efficiently produces butyric acid and is considered a promising host for anaerobic production of bulk chemicals. Due to limited knowledge on the genetic and metabolic characteristics of this strain, however, little progress has been made in metabolic engineering of this strain. Here we report the complete genome sequence of C. tyrobutyricum KCTC 5387 (ATCC 25755), which consists of a 3.07-Mbp chromosome and a 63-kbp plasmid. The results of genomic analyses suggested that C. tyrobutyricum produces butyrate from butyryl-coenzyme A (butyryl-CoA) through acetate reassimilation by CoA transferase, differently from Clostridium acetobutylicum, which uses the phosphotransbutyrylase-butyrate kinase pathway; this was validated by reverse transcription-PCR (RT-PCR) of related genes, protein expression levels, in vitro CoA transferase assay, and fed-batch fermentation. In addition, the changes in protein expression levels during the course of batch fermentations on glucose were examined by shotgun proteomics. Unlike C. acetobutylicum, the expression levels of proteins involved in glycolytic and fermentative pathways in C. tyrobutyricum did not decrease even at the stationary phase. Proteins related to energy conservation mechanisms, including Rnf complex, NfnAB, and pyruvate-phosphate dikinase that are absent in C. acetobutylicum, were identified. Such features explain why this organism can produce butyric acid to a much higher titer and better tolerate toxic metabolites. This study presenting the complete genome sequence, global protein expression profiles, and genome-based metabolic characteristics during the batch fermentation of C. tyrobutyricum will be valuable in designing strategies for metabolic engineering of this strain.
IMPORTANCE
Bio-based production of chemicals from renewable biomass has become increasingly important due to our concerns on climate change and other environmental problems. C. tyrobutyricum has been used for efficient butyric acid production. In order to further increase the performance and expand the capabilities of this strain toward production of other chemicals, metabolic engineering needs to be performed. For this, better understanding on the metabolic and physiological characteristics of this bacterium at the genome level is needed. This work reporting the results of complete genomic and proteomic analyses together with new insights on butyric acid biosynthetic pathway and energy conservation will allow development of strategies for metabolic engineering of C. tyrobutyricum for the bio-based production of various chemicals in addition to butyric acid.
Topics: Anaerobiosis; Butyric Acid; Clostridium tyrobutyricum; Fermentation; Gene Expression Profiling; Genome, Bacterial; Glucose; Metabolic Networks and Pathways; Plasmids; Proteome; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA
PubMed: 27302759
DOI: 10.1128/mBio.00743-16 -
Frontiers in Microbiology 2023has considerable prospect in the production of organic acids. Globally, refinery final molasses is rich in sugar and reported to have high levels of accumulation and...
INTRODUCTION
has considerable prospect in the production of organic acids. Globally, refinery final molasses is rich in sugar and reported to have high levels of accumulation and high emission costs, recognized as an excellent substrate for fermentation, but there is no suitable method available at present.
METHODS
In this study, an acid-base treatment combined with a new green membrane treatment technology - a dynamic ion-exchange membrane -was used to pretreat refinery final molasses, so that it could be used for to produce butyric acid. A high-performance liquid chromatography method was established to determine the conversion of a large amount of sucrose into fermentable sugars (71.88 g/L glucose and 38.06 g/L fructose) in the treated refinery final molasses. The process of sequential filtration with 3, 1, and 0.45 μm-pore diameter dynamic ion-exchange membranes could remove impurities, pigments, and harmful substances from the refinery final molasses, and retain the fermentable sugar.
RESULTS AND DISCUSSION
This means that refinery final molasses from the sugar industry could be utilized as a high-value by-product and used for the growth of , with industrial feasibility and economic competitiveness. Using the treated refinery final molasses as a carbon source, was screened by the method of adaptive evolution. The strain with butyric acid yielded 52.54 g/L, and the yield of the six carbon sugar was increased from 0.240 to 0.478 g/g. The results showed that combination of and ionic membrane technology broke through the bottleneck of its utilization of refinery final molasses. This study provided an innovative idea for the fermentation to produce butyric acid.
PubMed: 36825085
DOI: 10.3389/fmicb.2023.1065953 -
Food Research International (Ottawa,... Jan 2023Lytic bacteriophages (phages) offer a great potential as biocontrol agents for spoilage Clostridium tyrobutyricum, responsible for butyric acid fermentation in semi-hard...
Lytic bacteriophages (phages) offer a great potential as biocontrol agents for spoilage Clostridium tyrobutyricum, responsible for butyric acid fermentation in semi-hard and hard ripened cheeses, resulting in late gas blowing defect. With this aim, we have isolated, identified and characterized new lytic phages of C. tyrobutyricum, and have evaluated their efficacy to control cheese late blowing by adding them to manufacture milk. Silage, soil, milk and cheese from dairy farms were screened for anti-clostridial phages, obtaining 96 isolates active against C. tyrobutyricum. According to host range, source and plaque morphology, we obtained 20 phage profiles, 8 of them (represented by phages FA3, FA21, FA29, FA52, FA58, FA67, FA70 and FA88) showing a wider host range and high quality lysis, which were further characterized. Selected isolates showed a non-contractile tail, belonging to the Siphoviridae family, and were grouped into 3 restriction profiles. Viable phages were detected after storage in sodium-magnesium buffer (SM buffer), skim milk and acidified skim milk (pH 5) for 7 d at 4 °C, 12 °C and 37 °C, although a decline in infectivity was observed in some cases. Good phage survival was also detected during semi-hard cheese manufacture and ripening (60 d), and cheese lactococci counts, pH, dry matter values, and volatile compounds were not affected by phage addition. In semi-hard cheese, phage FA67 impaired the early germination of C. tyrobutyricum spores and caused a significant decrease in clostridial vegetative cells counts at 14 d of ripening, delaying by 2 weeks the consumption of lactic acid, formation of butyric acid and appearance of late blowing symptoms, compared to the spoilt control cheese without the phage. This is the first report on the application of phage to control C. tyrobutyricum in cheese.
Topics: Clostridium tyrobutyricum; Butyric Acid; Cheese; Bacteriophages; Clostridium
PubMed: 36596151
DOI: 10.1016/j.foodres.2022.112222 -
Frontiers in Microbiology 2022Probiotics contribute a lot to human health and the occurrence of diseases. Correspondingly, probiotics' safety evaluation and probiotic properties have received...
Probiotics contribute a lot to human health and the occurrence of diseases. Correspondingly, probiotics' safety evaluation and probiotic properties have received increasing attention in the food industry and disease treatment. L319 is a short-chain fatty acid (SCFA)-producing strain isolated from Grana Padano cheese with a blowing defect. Our previous study has shown its safety at the genomic level. This study focused more on the safety evaluation and probiotic properties . According to the results, this strain has no potential virulence factors or the possibility of antibiotic resistance genes propagation. It also fulfilled several criteria to be used as a probiotic, including significant hydrophobicity under an acidic condition (pH 5.0) and resistance to simulate gastric juice and intestinal juice. Additionally, this strain was found to be tolerant to the harsh conditions of the external environment, including resistance to low (20°C) and high (50°C) temperatures, high salts (3% NaCl), and low pH (pH 5.0). Finally, we found that this strain could ferment prebiotics, such as chito-oligosaccharides, to produce SCFAs. It exhibited excellent growth performance whether using chito-oligosaccharide as a sole carbon source or combining glucose as the mixed carbon source. Furthermore, chito-oligosaccharide and glucose (1:1) mixed carbon sources were the optimal strategy for the production of SCFAs. Our findings demonstrated that this strain might be considered a promising candidate for future use as a probiotic to promote health benefits.
PubMed: 35783414
DOI: 10.3389/fmicb.2022.926710 -
Nutrients Feb 2024Obesity and its complications constitute a main threat to global human health. The purpose of this investigation was to explore the influences of (Ct) on lipid...
Obesity and its complications constitute a main threat to global human health. The purpose of this investigation was to explore the influences of (Ct) on lipid metabolism, intestinal barrier function, and intestinal microbiome in obese mice induced by a high-fat diet (HFD). After establishing the obesity model, 10 CFU/mL and 10 CFU/mL were used to intervene in HFD-fed mice by gavage for six weeks, and indexes related to obesity were measured. In the liver of HFD-fed mice, the results revealed that reduced liver weight and the levels of triglyceride (TG), total cholesterol (TC), and nonesterified fatty acid (NEFA), along with decreasing red lipid droplets and fat vacuoles. After intervention, the mRNA expression of peroxisome proliferator-activated receptor-γ (PPARγ) was downregulated, and AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor-α (PPARα), adipose triglyceride lipase (ATGL), and hormone-sensitive lipase (HSL) were upregulated in the liver. Additionally, alleviated intestinal morphology injury caused by HFD, decreased the expression of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), and IL-1β in the colon, and upregulated tight junction protein expression. In addition, 16S rRNA sequencing revealed that increases the diversity of intestinal microbiota. Overall, improved HFD-induced lipid metabolism disorders, preserved the intestinal barrier's integrity, and modulated the structure of the intestinal microbiome. These findings provide a novel insight into the role of as a probiotic in regulating lipid metabolism.
Topics: Humans; Animals; Mice; Diet, High-Fat; Lipid Metabolism; Gastrointestinal Microbiome; Mice, Obese; Clostridium tyrobutyricum; Intestinal Barrier Function; RNA, Ribosomal, 16S; Peroxisome Proliferator-Activated Receptors; Liver; Obesity; Mice, Inbred C57BL
PubMed: 38398817
DOI: 10.3390/nu16040493 -
Biotechnology For Biofuels and... Jun 2022Butyric acid, an essential C4 platform chemical, is widely used in food, pharmaceutical, and animal feed industries. Clostridium tyrobutyricum is the most promising...
BACKGROUND
Butyric acid, an essential C4 platform chemical, is widely used in food, pharmaceutical, and animal feed industries. Clostridium tyrobutyricum is the most promising microorganism for industrial bio-butyrate production. However, the metabolic driving mechanism for butyrate synthesis was still not profoundly studied.
RESULTS
This study reports a first-generation genome-scale model (GEM) for C. tyrobutyricum, which provides a comprehensive and systematic analysis for the butyrate synthesis driving mechanisms. Based on the analysis in silico, an energy conversion system, which couples the proton efflux with butyryl-CoA transformation by two redox loops of ferredoxin, could be the main driving force for butyrate synthesis. For verifying the driving mechanism, a hydrogenase (HydA) expression was perturbed by inducible regulation and knockout. The results showed that HydA deficiency significantly improved the intracellular NADH/NAD rate, decreased acetate accumulation (63.6% in serum bottle and 58.1% in bioreactor), and improved the yield of butyrate (26.3% in serum bottle and 34.5% in bioreactor). It was in line with the expectation based on the energy conversion coupling driving mechanism.
CONCLUSIONS
This work show that the first-generation GEM and coupling metabolic analysis effectively promoted in-depth understanding of the metabolic driving mechanism in C. tyrobutyricum and provided a new insight for tuning metabolic flux direction in Clostridium chassis cells.
PubMed: 35752796
DOI: 10.1186/s13068-022-02169-z