ABSTRACT
Bacterial therapeutics are the emergent alternatives in treating autoimmune diseases such as Rheumatoid Arthritis [RA]. P. histicola MCI 001 is one such therapeutic bacterium that has been proven to treat autoimmune diseases such as RA and multiple sclerosis [MS] in animal models. The present study characterized P. histicola MCI 001 isolated from a human duodenal biopsy, and evaluated its impact on the gut microbial and metabolic profile in a longitudinal study using the collagen-induced arthritis model in HLA-DQ8.AEo transgenic mice. P. histicola MCI 001 though closely related to the type strain of P. histicola, DSM 19854, differed in utilizing glycerol. In culture, P. histicola MCI 001 produced vitamins such as biotin and folate, and was involved in digesting complex carbohydrates and production of acetate. Colonization study showed that duodenum was the predominant niche for the gavaged MCI 001. A longitudinal follow-up of gut microbial profile in arthritic mice treated with MCI 001 suggested that dysbiosis caused due to arthritis was partially restored to the profile of naïve mice after treatment. A taxon-level analysis suggested an expansion of intestinal genus Allobaculum in MCI001 treated arthritic mice. Eubiosis achieved post treatment with P. histicola MCI 001 was also reflected in the increased production of short-chain fatty acids [SCFAs]. Present study suggests that the treatment with P. histicola MCI 001 leads to an expansion of Allobaculum by increasing the availability of simple carbohydrates and acetate. Restoration of microbial profile and metabolites like butyrate induce immune and gut homeostasis.
Subject(s)
Biological Therapy/methods , Butyrates/metabolism , Prevotella/physiology , Symbiosis , Adaptation, Physiological , Animals , Arthritis, Rheumatoid/diagnosis , Arthritis, Rheumatoid/etiology , Arthritis, Rheumatoid/metabolism , Arthritis, Rheumatoid/therapy , Bile Acids and Salts/pharmacology , Disease Models, Animal , Fatty Acids, Volatile/metabolism , Gastric Juice , Gastrointestinal Microbiome , Humans , Hydrogen-Ion Concentration , Mice , Mice, Transgenic , Prevotella/classification , Prevotella/drug effects , Prevotella/geneticsABSTRACT
BACKGROUND: The human colon is colonised by a dense microbial community whose species composition and metabolism are linked to health and disease. The main energy sources for colonic bacteria are dietary polysaccharides and oligosaccharides. These play a major role in modulating gut microbial composition and metabolism, which in turn can impact on health outcomes. RESULTS: We investigated the influence of wheat bran arabinoxylan oligosaccharides (AXOS) and maltodextrin supplements in modulating the composition of the colonic microbiota and metabolites in healthy adults over the age of 60. Male and female volunteers, (n = 21, mean BMI 25.2 ± 0.7 kg/m2) participated in the double-blind, cross over supplement study. Faecal samples were collected for analysis of microbiota, short chain fatty acids levels and calprotectin. Blood samples were collected to measure glucose, cholesterol and triglycerides levels. There was no change in these markers nor in calprotectin levels in response to the supplements. Both supplements were well-tolerated by the volunteers. Microbiota analysis across the whole volunteer cohort revealed a significant increase in the proportional abundance of faecal Bifidobacterium species (P ≤ 0.01) in response to AXOS, but not maltodextrin, supplementation. There was considerable inter-individual variation in the other bacterial taxa that responded, with a clear stratification of volunteers as either Prevotella-plus (n = 8; > 0.1% proportional abundance) or Prevotella-minus (n = 13; ≤0.1% proportional abundance) subjects founded on baseline sample profiles. There was a significant increase in the proportional abundance of both faecal Bifidobacterium (P ≤ 0.01) and Prevotella species (P ≤ 0.01) in Prevotella-plus volunteers during AXOS supplementation, while Prevotella and Bacteroides relative abundances showed an inverse relationship. Proportional abundance of 26 OTUs, including bifidobacteria and Anaerostipes hadrus, differed significantly between baseline samples of Prevotella-plus compared to Prevotella-minus individuals. CONCLUSIONS: The wheat bran AXOS supplementation was bifidogenic and resulted in changes in human gut microbiota composition that depended on the initial microbiota profile, specifically the presence or absence of Prevotella spp. as a major component of the microbiota. Our data therefore suggest that initial profiling of individuals through gut microbiota analysis should be considered important when contemplating nutritional interventions that rely on prebiotics. TRIAL REGISTRATION: Clinical trial registration number: NCT02693782 . Registered 29 February 2016 - Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT02693782?term=NCT02693782&rank=1.
Subject(s)
Dietary Fiber , Gastrointestinal Microbiome/physiology , Oligosaccharides/pharmacology , Prevotella/physiology , Aged , Dietary Supplements , Double-Blind Method , Fatty Acids, Volatile/metabolism , Feces/chemistry , Feces/microbiology , Female , Gastrointestinal Microbiome/drug effects , Humans , Leukocyte L1 Antigen Complex/analysis , Lipids/blood , Male , Middle Aged , Oligosaccharides/chemistry , Polysaccharides/pharmacology , Prebiotics , Prevotella/drug effects , XylansABSTRACT
Our study demonstrated that sleep deprivation resulted in homeostasis disorder of colon. Our study goes deeper into the positive effects of melatonin on small intestinal microbiota disorder caused by sleep deprivation. We successfully established a multiplatform 72 h sleep deprivation mouse model with or without melatonin supplementation, and analyzed the change of small intestinal microbiota using high-throughput sequencing of the 16S rRNA. We found melatonin supplementation suppressed the decrease of plasma melatonin level in sleep deprivation mice. Meanwhile, melatonin supplementation improved significantly the reduction in OTU numbers and the diversity and richness of jejunal microbiota and the abundance of Bacteroidaeae and Prevotellaceae, as well as an increase in the Firmicutes-to-Bacteroidetes ratio and the content of Moraxellaceae and Aeromonadaceae in the jejunum of sleep deprived-mice. Moreover, melatonin supplementation reversed the change of metabolic pathway in sleep deprived-mice, including metabolism, signal transduction mechanisms and transcription etc, which were related to intestinal health. Furthermore, melatonin supplementation inverted the sleep deprivation-induced a decline of anti-inflammatory cytokines (IL-22) and an increase of the ROS and proinflammatory cytokines (IL-17) in jejunum. These findings suggested that melatonin, similar to a probiotics agent, can reverse sleep deprivation-induced small intestinal microbiota disorder by suppressing oxidative stress and inflammation response.
Subject(s)
Antioxidants/pharmacology , Dysbiosis/drug therapy , Gastrointestinal Microbiome/drug effects , Jejunum/microbiology , Melatonin/pharmacology , Sleep Deprivation/microbiology , Aeromonadaceae/drug effects , Aeromonadaceae/isolation & purification , Animals , Bacteroidaceae/drug effects , Bacteroidaceae/isolation & purification , Firmicutes/drug effects , Firmicutes/isolation & purification , Inflammation , Interleukin-17/analysis , Interleukins/analysis , Male , Mice , Mice, Inbred ICR , Moraxellaceae/drug effects , Moraxellaceae/isolation & purification , Oxidative Stress/drug effects , Prevotella/drug effects , Prevotella/isolation & purification , RNA, Ribosomal, 16S/genetics , Signal Transduction/drug effects , Transcription, Genetic/drug effects , Transcription, Genetic/genetics , Interleukin-22ABSTRACT
The polysaccharides and phenylethanoid glycosides from Cistanche deserticola have been demonstrated with various health benefits, however the interactive effect between these two kinds of compounds in vivo are not in detail known. The objective of this study was to investigate the synergistic actions of cistanche polysaccharides with phenylethanoid glycoside and the effects of polysaccharides on gut microbiota. Sprague-Dawley rats were fed with different kinds of cistanche polysaccharides for 20 days, on the last day, all rats were administered the echinacoside at 100 mg/kg. The results were compared mainly on the difference of pharmacokinetic parameters, gut microbiota composition, and short chain fatty acids contents. The results indicated that all the cistanche polysaccharides, including crude polysaccharide, high molecular weight polysaccharide and low molecular weight polysaccharide, could regulate the gut microbiota diversity, increase beneficial bacteria and particularly enhance the growth of Prevotella spp. as well as improve the production of short chain fatty acids and the absorption of echinacoside. By exploring the synergistic actions of polysaccharides with small molecules, these findings suggest that cistanche polysaccharides, particularly low molecular weight polysaccharides, could be used as a gut microbiota manipulator for health promotion.
Subject(s)
Cistanche/chemistry , Gastrointestinal Microbiome/drug effects , Glycosides/metabolism , Physical Phenomena , Plant Extracts/pharmacology , Polysaccharides/pharmacology , Animals , Fatty Acids, Volatile/metabolism , Male , Molecular Weight , Plant Extracts/chemistry , Polysaccharides/chemistry , Prevotella/drug effects , Prevotella/growth & development , RNA, Ribosomal, 16S , Rats , Rats, Sprague-DawleyABSTRACT
Some functional food components may help maintain homeostasis by promoting balanced gut microbiota. Here, we explore the possible complementary effects of d-fagomine and ω-3 polyunsaturated fatty acids (ω-3 PUFAs) eicosapentaenoic acid/docosahexaenoic acid (EPA/DHA 1:1) on putatively beneficial gut bacterial strains. Male Sprague-Dawley rats were supplemented with d-fagomine, ω-3 PUFAs, or both, for 23 weeks. Bacterial subgroups were evaluated in fecal DNA by quantitative real-time polymerase chain reaction (qRT-PCR) and short-chain fatty acids were determined by gas chromatography. We found that the populations of the genus Prevotella remained stable over time in animals supplemented with d-fagomine, independently of ω-3 PUFA supplementation. Animals in these groups gained less weight than controls and rats given only ω-3 PUFAs. d-Fagomine supplementation together with ω-3 PUFAs maintained the relative populations of Bacteroides. ω-3 PUFAs alone or combined with d-fagomine reduced the amount of acetic acid and total short-chain fatty acids in feces. The plasma levels of pro-inflammatory arachidonic acid derived metabolites, triglycerides and cholesterol were lower in both groups supplemented with ω-3 PUFAs. The d-fagomine and ω-3 PUFAs combination provided the functional benefits of each supplement. Notably, it helped stabilize populations of Prevotella in the rat intestinal tract while reducing weight gain and providing the anti-inflammatory and cardiovascular benefits of ω-3 PUFAs.
Subject(s)
Body Weight/drug effects , Fatty Acids, Omega-3/pharmacology , Gastrointestinal Microbiome/drug effects , Imino Pyranoses/pharmacology , Administration, Oral , Animals , Bacteroides/drug effects , Dietary Supplements , Fagopyrum/chemistry , Fatty Acids, Omega-3/administration & dosage , Imino Pyranoses/administration & dosage , Male , Prevotella/drug effects , Rats , Rats, Sprague-Dawley , SeafoodABSTRACT
Given the increasing prevalence of antibiotic resistance among bacterial strains and the side effects caused by synthetic drugs, it is increasingly important to investigate potential herbal alternatives. In the present study, antimicrobial, cell cytotoxicity, and cleaning tests were performed to evaluate the potential of Fufang Bingpeng irrigant as a root canal irrigant, in addition to q-PCR and high-throughput sequencing analyses. Our in vitro results showed a low minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Fufang Bingpeng irrigant against Porphyromonas gingivalis ATCC 33277 (6.25 and 12.5%, respectively), Prevotella intermedius ATCC 25611 (6.25 and 6.25%, respectively), Fusobacterium nucleatum ATCC 25286 (6.25 and 6.25%, respectively), Enterococcus faecalis ATCC 19433 (25 and 25%, respectively), and Bacteriodes fragilis ATCC 25285 (12.5 and 12.5%, respectively). Furthermore, it effectively removed the remaining debris and increased the number of open dentinal tubules in root canals compared to the NaCl irrigant (p < 0.05). Fufang Bingpeng irrigant also presented low cytotoxicity to L929 cells compared to the NaClO irrigant. The in vivo results indicated that all irrigants used significantly reduced the number of bacteria compared to the number prior to treatment, and only 1/104.95 bacteria remained in the root canal following the use of Fufang Bingpeng irrigant (p < 0.001). Moreover, the high-throughput sequencing results indicated that all irrigants markedly enhanced the α diversity in the root canal compared to the before preparation control group, while Fufang Bingpeng maintained better microbial diversity than other groups. Therefore, Fufang Bingpeng irrigant presents a promising alternative for use as a root canal irrigant in clinical settings.
Subject(s)
Anti-Infective Agents/administration & dosage , Root Canal Irrigants/administration & dosage , Sodium Hypochlorite/administration & dosage , Ultrasonics/methods , Adolescent , Cell Line/drug effects , Child , DNA, Bacterial/analysis , Dental Pulp Cavity/microbiology , Drugs, Chinese Herbal/administration & dosage , Enterococcus faecalis/drug effects , Fusobacterium nucleatum/drug effects , High-Throughput Screening Assays , Humans , Microbial Sensitivity Tests , Porphyromonas gingivalis/drug effects , Prevotella/drug effects , Root Canal Preparation , Root Canal Therapy , Ultrasonic TherapyABSTRACT
ABSTRACT Many essential oils (EOs) of different plant species possess interesting antimicrobial effects on buccal bacteria and antioxidant properties. Eugenia klotzschiana Berg (pêra-do-cerrado, in Portuguese) is a species of Myrtaceae with restricted distribution in the Cerrado. The essential oils were extracted through the hydrodistillation technique using a modified Clevenger apparatus (2 hours) and chemically characterized by GC-MS. The major compounds were α-copaene (10.6 %) found in oil from leaves in natura, β-bisabolene (17.4 %) in the essential oil from dry leaves and α-(E)-bergamotene (29.9 %) in oil from flowers. The antioxidant activity of essential oils showed similarities in both methods under analysis (DPPH and ABTS˙+) and the results suggested moderate to high antioxidant activity. The antibacterial activity was evaluated by determining minimum inhibitory concentrations (MICs), using the microdilution method. MIC values below 400 µg/mL were obtained against Streptococcus salivarius (200 µg/mL), S. mutans (50 µg/mL), S. mitis (200 µg/mL) and Prevotella nigrescens (50 µg/mL). This is the first report of the chemical composition and antibacterial and antioxidant activities of the essential oils of E. klotzschiana. These results suggest that E. klotzschiana, a Brazilian plant, provide initial evidence of a new and alternative source of substances with medicinal interest.
Subject(s)
Streptococcus/drug effects , Prevotella/drug effects , Plant Leaves/chemistry , Flowers/chemistry , Eugenia/chemistry , Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacology , Microbial Sensitivity Tests , Gas Chromatography-Mass Spectrometry , Anti-Bacterial Agents/isolation & purification , Antioxidants/isolation & purificationABSTRACT
Many essential oils (EOs) of different plant species possess interesting antimicrobial effects on buccal bacteria and antioxidant properties. Eugenia klotzschiana Berg (pêra-do-cerrado, in Portuguese) is a species of Myrtaceae with restricted distribution in the Cerrado. The essential oils were extracted through the hydrodistillation technique using a modified Clevenger apparatus (2 hours) and chemically characterized by GC-MS. The major compounds were α-copaene (10.6 %) found in oil from leaves in natura, ß-bisabolene (17.4 %) in the essential oil from dry leaves and α-(E)-bergamotene (29.9 %) in oil from flowers. The antioxidant activity of essential oils showed similarities in both methods under analysis (DPPH and ABTSË+) and the results suggested moderate to high antioxidant activity. The antibacterial activity was evaluated by determining minimum inhibitory concentrations (MICs), using the microdilution method. MIC values below 400 µg/mL were obtained against Streptococcus salivarius (200 µg/mL), S. mutans (50 µg/mL), S. mitis (200 µg/mL) and Prevotella nigrescens (50 µg/mL). This is the first report of the chemical composition and antibacterial and antioxidant activities of the essential oils of E. klotzschiana. These results suggest that E. klotzschiana, a Brazilian plant, provide initial evidence of a new and alternative source of substances with medicinal interest.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacology , Eugenia/chemistry , Flowers/chemistry , Plant Leaves/chemistry , Prevotella/drug effects , Streptococcus/drug effects , Anti-Bacterial Agents/isolation & purification , Antioxidants/isolation & purification , Gas Chromatography-Mass Spectrometry , Microbial Sensitivity TestsABSTRACT
This study investigated the effects of ensiled mulberry leaves (EML) and sun-dried mulberry fruit pomace (SMFP) on the ruminal bacterial and archaeal community composition of finishing steers. Corn grain- and cotton meal-based concentrate was partially replaced with EML or SMFP. The diets had similar crude protein (CP), neutral detergent fiber (NDF), and metabolizable energy. Following the feeding trial, the steers were slaughtered and ruminal liquid samples were collected to study the ruminal microbiome. Extraction of DNA, amplification of the V4 region of the 16S rRNA gene, and Illumina MiSeq pyrosequencing were performed for each sample. Following sequence de-noising, chimera checking, and quality trimming, an average of 209,610 sequences were generated per sample. Quantitative real-time PCR was performed to examine the selected bacterial species in the rumen. Our results showed that the predominant phyla were Bacteroidetes (43.90%), Firmicutes (39.06%), Proteobacteria (4.31%), and Tenericutes (2.04%), and the predominant genera included Prevotella (13.82%), Ruminococcus (2.51%), Butyrivibrio (2.38%), and Succiniclasticum (2.26%). Compared to the control group, EML and SMFP groups had a higher abundance of total bacteria (p < 0.001); however, the bacterial community composition was similar among the three groups. At the phylum level, there were no significant differences in Firmicutes (p = 0.7932), Bacteroidetes (p = 0.2330), Tenericutes (p = 0.2811), or Proteobacteria (p = 0.0680) levels among the three groups; however, Fibrobacteres decreased in EML (p = 0.0431). At the genus level, there were no differences in Prevotella (p = 0.4280), Ruminococcus (p = 0.2639), Butyrivibrio (p = 0.4433), or Succiniclasticum (p = 0.0431) levels among the groups. Additionally, the dietary treatments had no significant effects on the archaeal community composition in the rumen. Therefore, EML and SMFP supplementation had no significant effects on the ruminal bacterial or archaeal community composition of finishing steers.
Subject(s)
Archaea/drug effects , Bacteria/drug effects , Diet/veterinary , Gastrointestinal Microbiome/drug effects , Morus/chemistry , Plant Extracts/pharmacology , Rumen/microbiology , Animals , Archaea/isolation & purification , Bacteria/isolation & purification , Bacteroidetes/drug effects , Bacteroidetes/isolation & purification , Butyrivibrio/drug effects , Butyrivibrio/isolation & purification , Cattle , Firmicutes/drug effects , Firmicutes/isolation & purification , Fruit/chemistry , Plant Leaves/chemistry , Prevotella/drug effects , Prevotella/isolation & purification , Proteobacteria/drug effects , Proteobacteria/isolation & purification , Real-Time Polymerase Chain Reaction , Ruminococcus/drug effects , Ruminococcus/isolation & purification , Tenericutes/drug effects , Tenericutes/isolation & purificationABSTRACT
Chondroitin sulfate (CS) as a dietary supplement and a symptomatic slow acting (SYSA) drug has been used for years. Recently, CS has been demonstrated to be readily degraded and fermented in vitro by specific human gut microbes, hinting that dietary CS may pose a potential effect on gut microbiota composition in vivo. However, until now, little information is available on modulations of gut microbiota by CS. In the present study, modulations of gut microbiota in Kunming mice by CS and its oligosaccharide (CSO) were investigated by high-throughput sequencing. As evidenced by Heatmap and principal component analysis (PCA), the female microbiota were more vulnerable than the male microbiota to CS and CSO treatment. Besides, it is of interest to found that CS and CSO had differing effects on the abundance of Bacteroidales S24-7, Bacteroides, Helicobacter, Odoribacter, Prevotellaceae and Lactobacillus in male mice versus female mice. Collectively, we demonstrated a sex-dependent effect on gut microbiota of CS and CSO. In addition, since gut microbiota exerts a major effect on host physiology, our study highlighted that certain beneficial effects of CS may be associated with modulations of gut microbiota, which merits further investigation.
Subject(s)
Chondroitin Sulfates/administration & dosage , Gastrointestinal Microbiome/drug effects , Oligosaccharides/administration & dosage , Animals , Chondroitin Sulfates/chemistry , Female , Gastrointestinal Microbiome/genetics , Helicobacter pylori/drug effects , Helicobacter pylori/genetics , Host-Pathogen Interactions/drug effects , Humans , Male , Mice , Oligosaccharides/chemistry , Prevotella/drug effects , Prevotella/geneticsABSTRACT
Nonalcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease in developed countries. Recent studies indicated that the modification of gut microbiota plays an important role in the progression from simple steatosis to steatohepatitis. Epidemiological studies have demonstrated consumption of fish oil or perilla oil rich in n-3 polyunsaturated fatty acids (PUFAs) protects against NAFLD. However, the underlying mechanisms remain unclear. In the present study, we adopted 16s rRNA amplicon sequencing technique to investigate the impacts of fish oil and perilla oil on gut microbiomes modification in rats with high-fat diet- (HFD-) induced NAFLD. Both fish oil and perilla oil ameliorated HFD-induced hepatic steatosis and inflammation. In comparison with the low-fat control diet, HFD feeding significantly reduced the relative abundance of Gram-positive bacteria in the gut, which was slightly reversed by either fish oil or perilla oil. Additionally, fish oil and perilla oil consumption abrogated the elevated abundance of Prevotella and Escherichia in the gut from HFD fed animals. Interestingly, the relative abundance of antiobese Akkermansia was remarkably increased only in animals fed fish oil compared with HFD group. In conclusion, compared with fish oil, perilla oil has similar but slightly weaker potency against HFD-induced NAFLD and gut dysbiosis.
Subject(s)
Diet, High-Fat/adverse effects , Fish Oils/administration & dosage , Non-alcoholic Fatty Liver Disease/drug therapy , alpha-Linolenic Acid/administration & dosage , Animals , Dysbiosis/drug therapy , Dysbiosis/microbiology , Escherichia/drug effects , Escherichia/pathogenicity , Fatty Acids, Omega-3 , Female , Gram-Positive Bacteria/drug effects , Gram-Positive Bacteria/pathogenicity , High-Throughput Nucleotide Sequencing , Liver/drug effects , Liver/microbiology , Liver/pathology , Male , Non-alcoholic Fatty Liver Disease/pathology , Plant Oils/administration & dosage , Prevotella/drug effects , Prevotella/isolation & purification , Prevotella/pathogenicity , RNA, Ribosomal, 16S/genetics , RatsABSTRACT
Alterations in gut microbiota composition offer insights that may be relevant for several chronic conditions, including obesity. This study aimed to evaluate the effect of (-)-epigallocatechin gallate (EGCG) on the modulation of gut microbiota and biomarkers of colonic fermentation end-products in rats. Rats were fed an assigned diet of either a control diet, a 0.3% (w/w) EGCG diet, or a 0.6% (w/w) EGCG diet for 4 wk. Compared to the control group, the addition of 0.6% EGCG to the diet brought about a significant increase in the starch and protein contents in the feces collected in the fourth week of feeding, but the relative weights of abdominal adipose tissues of rats were inversely suppressed. Host-specific bacterial community composition, as determined by terminal restriction fragment length polymorphism (T-RFLP) patterns for fecal 16S ribosomal RNA, showed a significant response in the reduced occupation of Clostridium spp. and an increased trend of Bacteroides by dietary supplementation with EGCG. The 0.6% EGCG diet also influenced the status of Bifidobacterium and Prevotella to a lesser extent. Interestingly, the cecum of rats fed the 0.6% EGCG diet contained lower levels of acetic and butyric acids, whereas EGCG had little influence on the cecal level of propionic acid. EGCG also reduced the cecal p-cresol concentration in a dose-dependent fashion. In conclusion, dietary EGCG affects the growth of certain species of gut microbiota in rats and is associated with the cecal pattern of short chain fatty acids which could be responsible for regulating energy metabolism in the body.
Subject(s)
Biomarkers/metabolism , Catechin/analogs & derivatives , Dietary Supplements , Gastrointestinal Tract/drug effects , Gastrointestinal Tract/microbiology , Microbiota , Acetic Acid/metabolism , Animals , Bifidobacterium/drug effects , Bifidobacterium/isolation & purification , Butyric Acid/metabolism , Catechin/administration & dosage , Clostridium/drug effects , Clostridium/isolation & purification , Cresols/metabolism , Feces/microbiology , Fermentation , Male , Organ Size , Prevotella/drug effects , Prevotella/isolation & purification , Rats , Rats, Wistar , Weight Gain/drug effectsABSTRACT
BACKGROUND: In previous works we have shown that a low-molecular-mass (LMM) fraction from mushroom (Lentinus edodes) homogenate interferes with binding of Streptococcus mutans to hydroxyapatite and Prevotella intermedia to gingival cells. Additionally, inhibition of biofilm formation of both odonto- and periodonto-pathogenic bacteria and detachment from preformed biofilms have been described for this compound. Further purification of mushroom extract has been recently achieved and a sub-fraction (i.e. # 5) has been identified as containing the majority of the mentioned biological activities. The aim of this study was to characterise the bacterial receptors for the purified mushroom sub-fraction #5 in order to better elucidate the mode of action of this compound when interfering with bacterial adhesion to host surfaces or with bacteria-bacteria interactions in the biofilm state. METHODS: Candidate bacterial molecules to act as target of this compound were bacterial surface molecules involved in cell adhesion and biofilm formation, and, thus, we have considered cell wall associated proteins (CWPs), teichoic acid (TA) and lipoteichoic acid (LTA) of S. mutans, and outer membrane proteins (OMPs) and lipopolysaccharide (LPS) of P. intermedia. RESULTS: Fifteen S. mutans CWPs and TA were capable of binding sub-fraction #5, while LTA did not. As far as P. intermedia is concerned, we show that five OMPs interact with sub-fraction # 5. Capacity of binding to P. intermedia LPS was also studied but in this case negative results were obtained. CONCLUSIONS: Binding sub-fraction # 5 to surface molecules of S. mutans or P. intermedia may result in inactivation of their physiological functions. As a whole, these results indicate, at molecular level, the bacterial surface alterations affecting adhesion and biofim formation. For these antimicrobial properties, the compound may find use in daily oral hygiene.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteria/drug effects , Bacterial Adhesion/drug effects , Biological Products/pharmacology , Dental Caries/microbiology , Gingivitis/microbiology , Shiitake Mushrooms , Agaricales , Bacterial Proteins/metabolism , Biofilms/drug effects , Dental Caries/drug therapy , Gingivitis/drug therapy , Lipopolysaccharides/metabolism , Membrane Proteins/metabolism , Prevotella/drug effects , Streptococcus mutans/drug effects , Teichoic Acids/metabolismABSTRACT
BACKGROUND AND OBJECTIVE: There is a paucity of data in relation to the possible emergence of triclosan (TCS)-resistant bacteria following long-term exposure to TCS toothpaste. Therefore, this study investigated whether long-term continuous exposure to TCS in toothpaste selects for TCS-resistant bacteria within the oral biofilm. MATERIAL AND METHODS: Dental plaque samples were collected from 40 individuals during year 5 of a randomised controlled trial. Participants had been randomly assigned to use TCS (3000 µg/mL TCS) (n = 18) or placebo toothpaste (n = 22). Diluted plaque samples were plated on to Wilkins-Chalgren agar plates containing 5% (v/v) laked sheep red blood cells and TCS (concentrations ranging from 25 to 150 µg/mL) and incubated at 37 °C under microaerophilic and anaerobic conditions for 2-10 d. Selected bacterial isolates were identified by partial 16S rDNA sequencing and TCS minimum inhibitory concentration (MIC) determined for each isolate. RESULTS: At 3000 µg/mL TCS no growth was observed under microaerophilic or anaerobic conditions in either group. The MICs of TCS for all isolates ranged from 125 to 1000 µg/mL in both groups. Species common to both groups had similar MICs. Veillonella parvula and Campylobacter gracilis were the most frequent isolates from both groups, with similar MICs in both groups. CONCLUSION: The use of TCS-containing toothpaste did not appear to lead to an increase in MIC of TCS of oral bacterial isolates.
Subject(s)
Anti-Infective Agents, Local/therapeutic use , Bacteria/drug effects , Biofilms/drug effects , Drug Resistance, Bacterial , Toothpastes/therapeutic use , Triclosan/therapeutic use , Aggregatibacter actinomycetemcomitans/drug effects , Aggregatibacter actinomycetemcomitans/isolation & purification , Bacteria/classification , Bacteriological Techniques , Campylobacter/drug effects , Campylobacter/isolation & purification , Chronic Periodontitis/prevention & control , Dental Plaque/microbiology , Dental Plaque/prevention & control , Follow-Up Studies , Fusobacterium nucleatum/drug effects , Fusobacterium nucleatum/isolation & purification , Humans , Microbial Sensitivity Tests , Placebos , Porphyromonas gingivalis/drug effects , Porphyromonas gingivalis/isolation & purification , Prevotella/drug effects , Prevotella/isolation & purification , Streptococcus anginosus/drug effects , Streptococcus anginosus/isolation & purification , Streptococcus mutans/drug effects , Streptococcus mutans/isolation & purification , Veillonella/drug effects , Veillonella/isolation & purificationABSTRACT
BACKGROUND AND OBJECTIVE: Beta-lactam antibiotics prescribed in periodontal therapy are vulnerable to degradation by bacterial ß-lactamases. This study evaluated the occurrence of ß-lactamase-positive subgingival bacteria in chronic periodontitis subjects of USA origin, and assessed their in vitro resistance to metronidazole at a breakpoint concentration of 4 µg/mL. MATERIAL AND METHODS: Subgingival plaque specimens from deep periodontal pockets with bleeding on probing were removed from 564 adults with severe chronic periodontitis before treatment. The samples were transported in VMGA III and then plated onto: (i) nonselective enriched Brucella blood agar (EBBA) and incubated anaerobically for 7 d; and (ii) selective trypticase soy-bacitracin-vancomycin (TSBV) and incubated for 3 d in air + 5% CO2 . At the end of the incubation periods, the bacterial test species were identified and quantified. Specimen dilutions were also plated onto EBBA plates supplemented with 2 µg/mL of amoxicillin, a combination of 2 µg/mL of amoxicillin plus 2 µg/mL of the ß-lactamase inhibitor clavulanic acid, or 4 µg/mL of metronidazole, followed by anaerobic incubation for 7 d. Bacterial test species presumptively positive for ß-lactamase production were identified by growth on EBBA primary isolation plates supplemented with amoxicillin alone and no growth on EBBA primary isolation plates containing both amoxicillin plus clavulanic acid. A subset of such isolates was subjected to nitrocefin-based chromogenic disk testing to confirm the presence of ß-lactamase activity. In vitro resistance to 4 µg/mL of metronidazole was noted when growth of test species occurred on metronidazole-supplemented EBBA culture plates. RESULTS: Two-hundred and ninety-four (52.1%) of the study subjects yielded ß-lactamase-producing subgingival bacterial test species, with Prevotella intermedia/nigrescens, Fusobacterium nucleatum and other Prevotella species most frequently identified as ß-lactamase-producing organisms. Of the ß-lactamase-producing bacterial test species strains recovered, 98.9% were susceptible in vitro to metronidazole at 4 µg/mL. CONCLUSION: The occurrence of ß-lactamase-positive subgingival bacterial species in more than half of the subjects with severe chronic periodontitis raises questions about the therapeutic potential of single-drug regimens with ß-lactam antibiotics in periodontal therapy. The in vitro effectiveness of metronidazole against nearly all recovered ß-lactamase-producing subgingival bacterial species further supports clinical periodontitis treatment strategies involving the combination of systemic amoxicillin plus metronidazole.
Subject(s)
Chronic Periodontitis/microbiology , Gram-Negative Bacteria/enzymology , beta-Lactamases/biosynthesis , Adult , Aged , Aged, 80 and over , Amoxicillin/pharmacology , Amoxicillin-Potassium Clavulanate Combination/pharmacology , Anti-Bacterial Agents/pharmacology , Anti-Infective Agents/pharmacology , Bacteriological Techniques , Dental Plaque/microbiology , Drug Resistance, Bacterial , Enzyme Inhibitors/pharmacology , Female , Fusobacterium nucleatum/drug effects , Fusobacterium nucleatum/enzymology , Fusobacterium nucleatum/isolation & purification , Gingiva/microbiology , Gram-Negative Bacteria/drug effects , Gram-Negative Bacteria/isolation & purification , Humans , Male , Metronidazole/pharmacology , Middle Aged , Periodontal Pocket/microbiology , Prevotella/classification , Prevotella/drug effects , Prevotella/enzymology , Prevotella intermedia/drug effects , Prevotella intermedia/enzymology , Prevotella intermedia/isolation & purification , Prevotella nigrescens/drug effects , Prevotella nigrescens/enzymology , Prevotella nigrescens/isolation & purification , beta-Lactamase InhibitorsABSTRACT
AIMS: The objective of this study was to comprehensively evaluate quillaja (QSP) and yucca saponin (YSP) products with respect to their effects on diversity of rumen bacteria and archaea, abundance of selected microbes, and feed degradability and fermentation. METHODS AND RESULTS: Both QSP and YSP at doses 0-0.6 g l(-1) tended to increase degradability of feed substrate in in vitro rumen cultures, but to different extents. Neither one of the saponins affected the concentrations of ammonia, total volatile fatty acids, or molar proportion of acetate. However, QSP increased molar proportion of propionate and decreased that of butyrate, whereas YSP tended to decrease that of butyrate. As determined by qPCR, QSP and YSP did not affect the abundance of total bacteria or Ruminococcus albus. The QSP did not affect the abundances of Fibrobacter succinogenes or genus Prevotella, but tended to decrease that of Ruminococcus flavefaciens, whereas YSP significantly increased the abundance of R. flavefaciens and Prevotella, and numerically increased that of F. succinogenes. Both saponins increased archaeal abundance, although to small magnitudes (0.3-0.4 log). The protozoal populations were decreased significantly by QSP, but not by YSP. Based on DGGE and T-RFLP analysis, both saponins altered the bacterial community and species organization, but less so the archaeal community. CONCLUSIONS: This study demonstrated that saponins, although not effective in mitigating methane emission, may improve feed utilization at low doses, and modulate ruminal microbial communities in a dose-dependent manner. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study suggest that saponins at low doses may directly stimulate the growth of some rumen bacteria including cellulolytic bacteria, thus improving digestibility of feeds, independent of their defaunation activity. In contrast, saponins at high doses modulate rumen fermentation characteristically similar to defaunation.
Subject(s)
Archaea/drug effects , Bacteria/drug effects , Fermentation , Rumen/microbiology , Saponins/pharmacology , Ammonia/metabolism , Animal Feed , Animals , Biodiversity , Cattle , Fatty Acids, Volatile/analysis , Fatty Acids, Volatile/metabolism , Fibrobacter/drug effects , Methane/biosynthesis , Prevotella/drug effects , Quillaja/chemistry , Quillaja Saponins/pharmacology , Ruminococcus/drug effects , Yucca/chemistryABSTRACT
The compounds 2',6'-dihydroxy-4'-geranyloxyacetophenone (1) and 2',6'-dihydroxy-4'-farnesyloxy-acetophenone (2) are oxyprenylated secondary metabolites extracted from plants belonging to the Rutaceae family. In this study, 1 and 2 were synthesized and tested for their antimicrobial activity toward major oral pathogens. Compounds 1 and 2 were synthesized by selective prenylation of 2,4,6-trihydroxyacetophenone at the 4' position with geranyl and farnesyl bromide, respectively. Compound 1 showed stronger antimicrobial activity than 2 against major oral pathogens, including Gram positive bacteria (Streptococcus mutans, Streptococcus sobrinus), Gram negative bacteria (Prevotella intermedia, Porphyromonas gingivalis) and Candida albicans. Evidences were obtained that the mode of action of 1 and 2 may be related to their iron-chelating property. This study suggests that 1 and 2 may represent potential natural molecules for the prevention/treatment of common oral infections, including dental caries, periodontal disease, and candidiasis.
Subject(s)
Acetophenones/pharmacology , Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Mouth/microbiology , Plant Extracts/pharmacology , Rutaceae/chemistry , Acetophenones/chemical synthesis , Acetophenones/chemistry , Acetophenones/therapeutic use , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/therapeutic use , Antifungal Agents/chemical synthesis , Antifungal Agents/therapeutic use , Bacteria/drug effects , Candida albicans/drug effects , Candidiasis/drug therapy , Chelating Agents/chemical synthesis , Chelating Agents/pharmacology , Chelating Agents/therapeutic use , Dental Caries/drug therapy , Iron/metabolism , Microbial Sensitivity Tests , Periodontal Diseases/drug therapy , Phytotherapy , Plant Extracts/chemistry , Plant Extracts/therapeutic use , Porphyromonas/drug effects , Prenylation , Prevotella/drug effects , Streptococcus/drug effectsABSTRACT
Linalool and α-terpineol exhibited strong antimicrobial activity against periodontopathic and cariogenic bacteria. However, their concentration should be kept below 0.4 mg/ml if they are to be used as components of toothpaste or gargling solution. Moreover, other compounds with antimicrobial activity against periodontopathic and cariogenic bacteria should be used in combination.
Subject(s)
Anti-Bacterial Agents/pharmacology , Cyclohexenes/pharmacology , Monoterpenes/pharmacology , Oils, Volatile/pharmacology , Acyclic Monoterpenes , Anti-Bacterial Agents/toxicity , Cell Line, Tumor , Cell Survival/drug effects , Cyclohexane Monoterpenes , Cyclohexenes/toxicity , Dental Caries/microbiology , Drug Evaluation, Preclinical , Fusobacterium/drug effects , Humans , Microbial Sensitivity Tests , Monoterpenes/toxicity , Oils, Volatile/toxicity , Pasteurellaceae/drug effects , Periodontal Diseases/microbiology , Porphyromonas/drug effects , Prevotella/drug effects , Streptococcus mutans/drug effects , Streptococcus sobrinus/drug effectsABSTRACT
Control of dental plaque-related diseases has traditionally relied on non-specific removal of plaque by mechanical means. As our knowledge of oral disease mechanisms increases, future treatment is likely to be more targeted, for example at small groups of organisms, single species or at key virulence factors they produce. The aim of this review is to consider the current status as regards novel treatment approaches. Maintenance of oral hygiene often includes use of chemical agents; however, increasing problems of resistance to synthetic antimicrobials have encouraged the search for alternative natural products. Plants are the source of more than 25% of prescription and over-the-counter preparations, and the potential of natural agents for oral prophylaxis will therefore be considered. Targeted approaches may be directed at the black-pigmented anaerobes associated with periodontitis. Such pigments provide an opportunity for targeted phototherapy with high-intensity monochromatic light. Studies to date have demonstrated selective killing of Porphyromonas gingivalis and Prevotella intermedia in biofilms. Functional inhibition approaches, including the use of protease inhibitors, are also being explored to control periodontitis. Replacement therapy by which a resident pathogen is replaced with a non-pathogenic bacteriocin-producing variant is currently under development with respect to Streptococcus mutans and dental caries.
Subject(s)
Anti-Bacterial Agents , Dental Plaque/microbiology , Dental Plaque/prevention & control , Periodontal Diseases/drug therapy , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Bacteroidaceae Infections/drug therapy , Bacteroidaceae Infections/microbiology , Humans , Light , Myrtaceae/chemistry , Periodontal Diseases/microbiology , Periodontal Diseases/prevention & control , Plant Oils/therapeutic use , Porphyromonas gingivalis/drug effects , Prevotella/drug effects , Probiotics , Protease Inhibitors/therapeutic use , Treatment OutcomeABSTRACT
The purpose of this study is to formulate in situ implants containing doxycycline hydrochloride and/or secnidazole that could be used in the treatment of periodontitis by direct periodontal intrapocket administration. Biodegradable polymers [poly (lactide) (PLA) and poly (lactide-co-glycolide) (PLGA)], each polymer in two concentrations 25%w/w, 35%w/w were used to formulate the in situ implants. The rheological behavior, in vitro drug release and the antimicrobial activity of the prepared implants were evaluated. Increasing the concentration of each polymer increases the viscosity and decreases the percent of the drugs released after 24 h. PLA implants showed a slower drugs release rate than PLGA implants in which the implants composed of 25% PLGA showed the fastest drugs release. The in vitro drug release and antimicrobial activity results were compared with results of Atridox. Results revealed that the pharmaceutical formulation based on 25% PLGA containing secnidazole and doxycycline hydrochloride has promising activity in treating periodontitis in comparison with Atridox.