RESUMEN
Biliary dysbiosis is associated with gallbladder cancer (GBC). We aimed to look for biliary bacteria specifically detected in GBC patients. We used 16S rRNA-based metagenomic analysis to elucidate biliary microbiota in 30 GBC and 30 gallstones-associated chronic cholecystitis patients. Relative abundance of five genera, Streptococcus, Enterococcus, Halomonas, Escherichia and Caulobacter was significantly associated with GBC. Of 15-species, 7 were detected significantly higher in GBC, Streptococcus anginosus, Streptococcus constellatus, Streptococcus intermedius, Actinomyces bowdenii, Actinomyces israelii, Actinomyces gerencseriae, and Escherichia fergusonii were biosafety level-2 infectious bacteria; other 8 species were biosafety level-1 bacteria. These bacterial species may be involved in pathogenesis of GBC.
Asunto(s)
Colecistitis , Neoplasias de la Vesícula Biliar , Cálculos Biliares , Metagenómica , Humanos , Neoplasias de la Vesícula Biliar/microbiología , Colecistitis/microbiología , Cálculos Biliares/microbiología , Femenino , Masculino , Metagenómica/métodos , Persona de Mediana Edad , Anciano , ARN Ribosómico 16S/genética , Enfermedad Crónica , Bacterias/genética , Bacterias/clasificación , Bacterias/aislamiento & purificación , Microbiota , Adulto , Disbiosis/microbiologíaRESUMEN
BACKGROUND: The formation of gallstones is often accompanied by chronic inflammation, and the mechanisms underlying inflammation and stone formation are not fully understood. Our aim is to utilize single-cell transcriptomics, bulk transcriptomics, and microbiome data to explore key pathogenic bacteria that may contribute to chronic inflammation and gallstone formation, as well as their associated mechanisms. METHODS: scRNA-seq data from a gallstone mouse model were extracted from the Gene Expression Omnibus (GEO) database and analyzed using the FindCluster() package for cell clustering analysis. Bulk transcriptomics data from patients with gallstone were also extracted from the GEO database, and intergroup functional differences were assessed using GO and KEGG enrichment analysis. Additionally, 16S rRNA sequencing was performed on gallbladder mucosal samples from asymptomatic patients with gallstone (n = 6) and liver transplant donor gallbladder mucosal samples (n = 6) to identify key bacteria associated with stone formation and chronic inflammation. Animal models were constructed to investigate the mechanisms by which these key pathogenic bacterial genera promote gallstone formation. RESULTS: Analysis of scRNA-seq data from the gallstone mouse model (GSE179524) revealed seven distinct cell clusters, with a significant increase in neutrophil numbers in the gallstone group. Analysis of bulk transcriptomics data from patients with gallstone (GSE202479) identified chronic inflammation in the gallbladder, potentially associated with dysbiosis of the gallbladder microbiota. 16S rRNA sequencing identified Helicobacter pylori as a key bacterium associated with gallbladder chronic inflammation and stone formation. CONCLUSIONS: Dysbiosis of the gallbladder mucosal microbiota is implicated in gallstone disease and leads to chronic inflammation. This study identified H. pylori as a potential key mucosal resident bacterium contributing to gallstone formation and discovered its key pathogenic factor CagA, which causes damage to the gallbladder mucosal barrier. These findings provide important clues for the prevention and treatment of gallstones.
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Antígenos Bacterianos , Proteínas Bacterianas , Células Epiteliales , Vesícula Biliar , Cálculos Biliares , Helicobacter pylori , Animales , Cálculos Biliares/microbiología , Cálculos Biliares/patología , Células Epiteliales/microbiología , Ratones , Humanos , Vesícula Biliar/microbiología , Vesícula Biliar/patología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Antígenos Bacterianos/genética , Antígenos Bacterianos/metabolismo , Helicobacter pylori/genética , Helicobacter pylori/patogenicidad , Helicobacter pylori/fisiología , ARN Ribosómico 16S/genética , Modelos Animales de Enfermedad , Permeabilidad , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/patología , Femenino , Masculino , Ratones Endogámicos C57BLRESUMEN
Introduction: The incidence of biliary system diseases has been continuously increasing in the past decade. Biliary system diseases bring a heavy burden to humanity and society. However, the specific etiology and pathogenesis are still unknown. The biliary system, as a bridge between the liver and intestine, plays an indispensable role in maintaining the physiological metabolism of the body. Therefore, prevention and treatment of biliary diseases are crucial. It is worth noting that the microorganisms participate in the lipid metabolism of the bile duct, especially the largest proportion of intestinal bacteria. Methods: We systematically reviewed the intestinal microbiota in patients with gallstones (GS), non-calculous biliary inflammatory, and biliary tract cancer (BTC). And searched Pubmed, Embase and Web of science for research studies published up to November 2023. Results: We found that the abundance of Faecalibacterium genus is decreased in GS, primary sclerosing cholangitis (PSC), primary biliary cholangitis (PBC) and BTC. Veillonella, Lactobacillus, Streptococcus and Enterococcus genus were significantly increased in PSC, PBC and BTC. Interestingly, we found that the relative abundance of Clostridium was generally reduced in GS, PBC and BTC. However, Clostridium was generally increased in PSC. Discussion: The existing research mostly focuses on exploring the mechanisms of bacteria targeting a single disease. Lacking comparison of multiple diseases and changes in bacteria during the disease process. We hope to provide biomarkers forearly diagnosis of biliary system diseases and provide new directions for the mechanism of intestinal microbiota in biliary diseases.
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Enfermedades de las Vías Biliares , Microbioma Gastrointestinal , Humanos , Enfermedades de las Vías Biliares/microbiología , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacterias/metabolismo , Cálculos Biliares/microbiología , Faecalibacterium , Lactobacillus , Sistema Biliar/microbiología , Neoplasias del Sistema Biliar/microbiología , Clostridium/aislamiento & purificación , Colangitis Esclerosante/microbiología , Enterococcus , Streptococcus/aislamiento & purificaciónRESUMEN
BACKGROUND: Gallstone disease is a common health problem worldwide. The role of the gut microbiota in gallstone pathogenesis remains obscure. Our aim was to evaluate the association and crosstalk between gut microbiota, gut metabolomic, and metabolic parameters in cholesterol gallstone patients, pigmented gallstone patients, and controls. METHODS: We collected stool samples from healthy individuals and patients with gallstones in our hospital from March 2019 to February 2021. 16s rRNA sequencing was performed, followed by differential abundance analyses. Measurement of bile acids and short-chain fatty acids was conducted via targeted metabolomics. RESULT: Thirty healthy individuals and 20 gallstone patients were recruited. The intergroup difference of microbial composition was significant between control and gallstone patients. The control group had more abundant Faecalibacterium , Prevotella 9 , and Bacteroides plebeius DSM 17135 . The cholesterol stones group had higher Desulfovibrionaceae and Bacteroides uniformis than the other two groups, while the pigment stone group had more abundant Escherichia-Shigella . In the analysis of metabolites, only n-butyric acid had a significantly higher concentration in the controls than in the gallstone group ( p < 0.01). The level of 3α-hydroxy-12 ketolithocholic acid, deoxycholic acid, and cholic acid showed no intergroup differences but was correlated to the serum cholesterol level and bacterial richness and evenness. CONCLUSION: Our study revealed the key taxa that can discriminate between individuals with or without gallstones. We also identified metabolites that are possibly associated with metabolic parameter and bacterial diversity. However, the correlation of the metabolites to certain clusters of bacteria should be analyzed in a larger cohort.
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Heces , Cálculos Biliares , Microbioma Gastrointestinal , Humanos , Cálculos Biliares/microbiología , Heces/microbiología , Heces/química , Persona de Mediana Edad , Femenino , Masculino , Adulto , Anciano , Metaboloma , Taiwán , Ácidos y Sales Biliares/metabolismo , Ácidos y Sales Biliares/análisis , ARN Ribosómico 16S/análisisRESUMEN
Gallstones are crystalline deposits in the gallbladder that are traditionally classified as cholesterol, pigment, or mixed stones based on their composition. Microbiota and host metabolism variances among the different types of gallstones remain largely unclear. Here, the bile and gallstone microbial species spectra of 29 subjects with gallstone disease (GSD, 24 cholesterol and 5 pigment) were revealed by type IIB restriction site-associated DNA microbiome sequencing (2bRAD-M). Among them (21 subjects: 18 cholesterol and 3 pigment), plasma samples were subjected to liquid chromatography-mass spectrometry (LC-MS) untargeted metabolomics. The microbiome yielded 896 species comprising 882 bacteria, 13 fungi, and 1 archaeon. Microbial profiling revealed significant enrichment of Cutibacterium acnes and Microbacterium sp005774735 in gallstone and Agrobacterium pusense and Enterovirga sp013044135 in the bile of cholesterol GSD subjects. The metabolome revealed 2296 metabolites, in which malvidin 3-(6''-malonylglucoside), 2-Methylpropyl glucosinolate, and ergothioneine were markedly enriched in cholesterol GSD subjects. Metabolite set enrichment analysis (MSEA) demonstrated enriched bile acids biosynthesis in individuals with cholesterol GSD. Overall, the multi-omics analysis revealed that microbiota and host metabolism interaction perturbations differ depending on the disease type. Perturbed gallstone type-related microbiota may contribute to unbalanced bile acids metabolism in the gallbladder and host, representing a potential early diagnostic marker and therapeutic target for GSD.
Asunto(s)
Cálculos Biliares , Humanos , Cálculos Biliares/química , Cálculos Biliares/metabolismo , Cálculos Biliares/microbiología , Ácidos y Sales Biliares/análisis , Bilis/química , Bilis/metabolismo , Colesterol/metabolismoRESUMEN
Gallstones are a prevalent biliary system disorder that is particularly common in women. They can lead to various complications, such as biliary colic, infection, cholecystitis, and even gallbladder cancer. However, the etiology of gallstones remains incompletely understood. The significant role of bacteria in gallstone formation has been demonstrated in recent studies. Certain bacteria not only influence bile composition and the gallbladder environment but also actively participate in stone formation by producing enzymes such as ß-glucuronidase and mucus. Therefore, this review aimed to analyze the mechanisms involving the types and quantities of bacteria involved in gallstone formation, providing valuable references for understanding the etiology and clinical treatment of gallstones.
Asunto(s)
Cálculos Biliares , Femenino , Humanos , Cálculos Biliares/microbiología , Bacterias/genética , Bilis/microbiologíaRESUMEN
INTRODUCTION: Salmonellosis, which is typically distinguished by an immediate onset of fever, abdominal pain, diarrhea, nausea, and vomiting, is a bacterial infection caused by Salmonella. The rising incidence of antibiotic resistance in Salmonella Typhimurium is a major worldwide problem, and a better knowledge of the distribution of antibiotic resistance patterns in Salmonella Typhimurium is critical for selecting the best antibiotic for infection treatment. In this work, the efficiency of bacteriophage therapy of vegetative cells and biofilms of S. Typhimurium was investigated. METHODS: Based on their host ranges, five Bacteriophages were chosen for therapy against 22 Salmonella isolates collected from various sources. PSCs1, PSDs1, PSCs2, PSSr1, and PSMc1 phages were found to exhibit potent anti-S. Typhimurium properties. In a 96-well microplate, the efficacy of bacteriophage therapy (105-1011 PFU/mL) against S. Typhimurium biofilm formers was first tested. A bacteriophage treatment (109 PFU/mL) was subsequently applied in the laboratory for 24 hours to minimize Salmonella adhering to the surfaces of gallstones and teeth. In 96-well microplate experiments, bacteriophage treatment inhibited biofilm development and reduced biofilm by up to 63.6% (P ≤ 0.05). RESULT: When compared to controls, bacteriophages (PSCs1, PSDs1, PSCs2, PSSr1, PSMc1) demonstrated a rapid drop in the populations of S. Typhimurium biofilms generated on the surfaces of gallstones and teeth where the structure of the Salmonella bacteria in the biofilm was broken and holes were created. CONCLUSION: Clearly, this study indicated that phages might be employed to eliminate S. Typhimurium biofilms on gallstone and tooth surfaces.
Asunto(s)
Bacteriófagos , Cálculos Biliares , Humanos , Cálculos Biliares/microbiología , Biopelículas , Salmonella typhimurium , Antibacterianos/farmacologíaRESUMEN
Introduction: Cholelithiasis has a high incidence worldwide and limited treatment options due to its poorly understood pathogenesis. Furthermore, the role of biliary microbiota in cholelithiasis remains understudied. To address these questions, we performed microbial sequencing from biliary samples from primary bile duct stone (PBDS) and secondary bile duct stone (SBDS) patients. Results: We analyzed in total 45 biliary samples, including those from cholelithiasis patients with PBDS or SBDS and people with other digestive diseases. 16S rRNA sequencing showed the bacteria family Alcaligenaceae increased in relative abundance in the lithiasis group compared with the non-lithiasis group. In addition, the PBDS group showed significantly lower bacterial diversity than SBDS, with Propionibacteriaceae, Sphingomonadaceae, and Lactobacillaceae as the most significant bacteria families decreased in relative abundance. We further performed whole metagenomic shotgun sequencing (wMGS) and found increased ability of biofilm synthesis and the ability to sense external stimuli in PBDS based on functional annotation of mapped reads. From genome-resolved analysis of the samples, we identified 36 high-quality draft bacterial genome sequences with completion ≥70% and contamination ≤10%. Most of these genomes were classified into Proteobacteria, Firmicutes, or Actinobacteria. Conclusions: Our findings indicated that there is a subtle impact on biliary microbiome from cholelithiasis while the difference is more pronounced between the PBDS and SBDS. It was revealed that the diversity of biliary microbiota in PBDS is lower, while some metabolic pathways are up-regulated, including those linked to higher incidence of different types of cancer, providing new insights for the understanding of cholelithiasis with different origin.
Asunto(s)
Cálculos Biliares , Microbiota , Bacterias/genética , Cálculos Biliares/genética , Cálculos Biliares/microbiología , Humanos , Metagenoma , ARN Ribosómico 16S/genéticaRESUMEN
Background: Hepatic lipid metabolism regulates biliary composition and influences the formation of cholesterol gallstones. The genes Hmgcr and Cyp7a1, which encode key liver enzymes, are regulated by circadian rhythm-related transcription factors. We aimed to investigate the effect of circadian rhythm disruption on hepatic cholesterol and bile acid metabolism and the incidence of cholesterol stone formation. Methods: Adult male C57BL/6J mice were fed either a lithogenic diet (LD) only during the sleep phase (time-restricted lithogenic diet feeding, TRF) or an LD ad libitum (non-time-restricted lithogenic diet feeding, nTRF) for 4 weeks. Food consumption, body mass gain, and the incidence of gallstones were assessed. Circulating metabolic parameters, lipid accumulation in the liver, the circadian expression of hepatic clock and metabolic genes, and the gut microbiota were analyzed. Results: TRF caused a dysregulation of the circadian rhythm in the mice, characterized by significant differences in the circadian expression patterns of clock-related genes. In TRF mice, the circadian rhythms in the expression of genes involved in bile acid and cholesterol metabolism were disrupted, as was the circadian rhythm of the gut microbiota. These changes were associated with high biliary cholesterol content, which promoted gallstone formation in the TRF mice. Conclusion: Disordered circadian rhythm is associated with abnormal hepatic bile acid and cholesterol metabolism in mice, which promotes gallstone formation.
Asunto(s)
Trastornos Cronobiológicos/complicaciones , Cálculos Biliares/etiología , Microbioma Gastrointestinal , Metabolismo de los Lípidos , Hígado/metabolismo , Animales , Colesterol/metabolismo , Trastornos Cronobiológicos/etiología , Trastornos Cronobiológicos/metabolismo , Trastornos Cronobiológicos/microbiología , Ritmo Circadiano/fisiología , Dieta/efectos adversos , Cálculos Biliares/metabolismo , Cálculos Biliares/microbiología , Microbioma Gastrointestinal/fisiología , Metabolismo de los Lípidos/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Factores de TiempoRESUMEN
Salmonella enterica serovar Typhi causes chronic infections by establishing biofilms on cholesterol gallstones. The production of extracellular polymeric substances (EPSs) is key to biofilm development, and biofilm architecture depends on which EPSs are made. The presence and spatial distribution of Salmonella EPSs produced in vitro and in vivo were investigated in Salmonella enterica serovar Typhimurium and S. Typhi biofilms by confocal microscopy. Comparisons between serovars and EPS-mutant bacteria were carried out by examining growth on cholesterol-coated surfaces, with human gallstones in ox or human bile, and in mice with gallstones. On cholesterol-coated surfaces, no major differences in EPS biomass were found between serovars. Cocultured biofilms containing wild-type (WT) and EPS-mutant bacteria demonstrated WT compensation for EPS mutations. Analysis of biofilm EPSs from gallbladder-mimicking conditions found that culture in human bile more consistently replicated the relative abundance and spatial organization of each EPS on gallstones from the chronic mouse model than culture in ox bile. S. Typhimurium biofilms cultured in vitro on gallstones in ox bile exhibited colocalized pairings of curli fimbriae/lipopolysaccharide and O-antigen capsule/cellulose, while these associations were not present in S. Typhi biofilms or in mouse gallstone biofilms. In general, the inclusion of human bile with gallstones in vitro replicated biofilm development on gallstones in vivo, demonstrating the strength of this model for studying biofilm parameters or EPS-directed therapeutic treatments.
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Biopelículas/crecimiento & desarrollo , Matriz Extracelular de Sustancias Poliméricas/fisiología , Cálculos Biliares/microbiología , Salmonella typhi/fisiología , Salmonella typhimurium/fisiología , Animales , Colesterol/metabolismo , Femenino , Humanos , RatonesRESUMEN
Extrahepatic cholangiocarcinoma (CCA) accounts for 3% of digestive cancers. The role of biliary microbiota as an environment-related modulator has been scarcely investigated in CCA, and the putative impact of associated diseases has not been yet assessed. We characterized the biliary microbiota in CCA patients in order to identify a specific CCA-related dysbiosis. The biliary effluents were collected through an endoscopic retrograde pancreatic cholangiography (ERCP) examination involving 28 CCA and 47 patients with gallstones, herein considered as controls. The biliary effluents were submitted to bacterial DNA extraction and 16S rRNA sequencing, using Illumina technology. Overall, 32% of CCA and 22% of controls displayed another associated disease, such as diabetes, pancreatitis, inflammatory bowel disease, or primary sclerosing cholangitis. Such associated diseases were considered in the comparisons that were made. Principal coordinate analysis (PCoA) detected a significant disparity of biliary microbiota composition between CCA patients and controls without an associated disease. Amongst the most abundant phyla, Proteobacteria did not significantly differ between CCA patients and controls, whereas Firmicutes levels were lower and Bacteroidetes higher in CCAs' biliary microbiota than in the controls' microbiota. The most abundant genera were Enterococcus, Streptococcus, Bacteroides, Klebsiella, and Pyramidobacter in CCA's biliary microbiota. Additionally, levels of Bacteroides, Geobacillus, Meiothermus, and Anoxybacillus genera were significantly higher in CCA patients' biliary microbiota, without an associated disease, in comparison with controls. A specific CCA-related dysbiosis was identified as compared to controls independently from associated diseases. This suggests that a microorganism community may be involved in CCA pathogenesis.
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Neoplasias de los Conductos Biliares/microbiología , Colangiocarcinoma/microbiología , Disbiosis/microbiología , Microbiota , Adulto , Anciano , Bacteroidetes/aislamiento & purificación , Bilis/microbiología , Neoplasias de los Conductos Biliares/complicaciones , Colangiocarcinoma/complicaciones , Disbiosis/complicaciones , Femenino , Firmicutes/aislamiento & purificación , Cálculos Biliares/microbiología , Humanos , Masculino , Persona de Mediana Edad , Proteobacteria/aislamiento & purificaciónRESUMEN
Cholelithiasis with chronic cholecystitis is prevalent and threatens human health. Most cholecystitis caused by bacterial infection or biofilms is accompanied by gallstones in the clinic, making gallbladder removal the only effective solution. Here, we provide a strategy to eliminate gallstone biofilms and dissolve gallstones by oral administration of a supernatant derived from nanoscale iron sulfide (nFeS supernatant). First, by using gallstones obtained from the clinic, we simulated biofilm formation on gallstones and tested the antibacterial activity of a nFeS supernatant in vitro. We found that the supernatant kills bacteria with a 5-log reduction in viability and destroys the biofilm structure. Smashed gallstones coincubated with E. coli biofilms promote gallstone formation, while nFeS supernatant can inhibit this process. Second, by using a murine (C57BL/6) model of cholelithiasis and cholecystitis, we tested the antibacterial efficacy and therapeutic effects of nFeS supernatant on cholelithiasis in vivo. Animal experimental data show that oral administration of nFeS supernatant can reduce 60% of bacteria in the gallbladder and, remarkably, remove gallstones with 2 days of treatment compared with clinical drug combinations (chenodeoxycholid acid and ciprofloxacin). Third, by performing protein abundance analysis of L02 cells and mouse livers, we observed the changes in CYP7a1, HMGCR, and SCP2 expression, indicating that the nFeS supernatant can also regulate cholesterol metabolism to prevent gallstone formation. Finally, hematologic biochemistry analysis and high-throughput sequencing technology show that the nFeS supernatant possesses high biocompatibility. Therefore, our work demonstrates that the nFeS supernatant may be a potential regimen for the treatment of cholelithiasis and cholecystitis by oral administration.
Asunto(s)
Antibacterianos/farmacología , Materiales Biocompatibles/farmacología , Colecistitis/tratamiento farmacológico , Compuestos Ferrosos/farmacología , Cálculos Biliares/tratamiento farmacológico , Nanopartículas/química , Administración Oral , Animales , Antibacterianos/administración & dosificación , Materiales Biocompatibles/administración & dosificación , Biopelículas/efectos de los fármacos , Línea Celular , Colecistitis/microbiología , Enfermedad Crónica , Modelos Animales de Enfermedad , Escherichia coli/efectos de los fármacos , Compuestos Ferrosos/administración & dosificación , Cálculos Biliares/microbiología , Humanos , Masculino , Ensayo de Materiales , Ratones , Ratones Endogámicos C57BL , Pruebas de Sensibilidad Microbiana , Nanopartículas/administración & dosificación , Tamaño de la PartículaRESUMEN
BACKGROUND: Cholesterol gallstones account for over 80% of gallstones, and the pathogenesis of gallstone formation involves genetic and environmental factors. However, data on the evolution of cholesterol gallstones with various densities are limited. This study aimed to determine the roles of microbiota and mucins on the formation of calcified cholesterol gallstones in patients with cholelithiasis. METHODS: Paired gallbladder tissues and bile specimens were obtained from cholelithiasis patients who were categorized into the isodense group and calcified group according to the density of gallstones. The relative abundance of microbiota in gallbladder tissues was detected. Immunohistochemistry and enzyme-linked immunosorbent assay were performed to detect the expression levels of MUC1, MUC2, MUC3a, MUC3b, MUC4, MUC5ac and MUC5b in gallbladder tissues and bile. The correlation of microbiota abundance with MUC4 expression was evaluated by linear regression. RESULTS: A total of 23 patients with gallbladder stones were included. The density of gallstones in the isodense group was significantly lower than that of the calcified group (34.20 ± 1.50 vs. 109.40 ± 3.84 HU, P < 0.0001). Compared to the isodense group, the calcified group showed a higher abundance of gram-positive bacteria at the fundus, in the body and neck of gallbladder tissues. The concentrations of MUC1, MUC2, MUC3a, MUC3b, MUC5ac and MUC5b in the epithelial cells of gallbladder tissues showed no difference between the two groups, while the concentrations of MUC4 were significantly higher in the calcified group than that in the isodense group at the fundus (15.49 ± 0.69 vs. 10.23 ± 0.54 ng/mL, P < 0.05), in the body (14.54 ± 0.94 vs. 11.87 ± 0.85 ng/mL, P < 0.05) as well as in the neck (14.77 ± 1.04 vs. 10.85 ± 0.72 ng/mL, P < 0.05) of gallbladder tissues. Moreover, the abundance of bacteria was positively correlated with the expression of MUC4 (r = 0.569, P < 0.05) in the calcified group. CONCLUSIONS: This study showed the potential clinical relevance among biliary microbiota, mucins and calcified gallstones in patients with gallstones. Gram-positive microbiota and MUC4 may be positively associated with the calcification of cholesterol gallstones.
Asunto(s)
Bilis/microbiología , Calcinosis/clasificación , Colesterol/metabolismo , Cálculos Biliares/clasificación , Regulación de la Expresión Génica , Microbiota , Mucina 4/genética , Adulto , Bilis/metabolismo , Calcinosis/genética , Calcinosis/microbiología , Femenino , Vesícula Biliar/microbiología , Cálculos Biliares/genética , Cálculos Biliares/microbiología , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Mucina 4/biosíntesis , ARN/genética , Estudios RetrospectivosRESUMEN
AIM: In our previous study, we have isolated different genera of bacteria from gallstones and this intrigued us to study their role in gallstone formation. The isolates exhibited certain biliary activities like urease activity, slime production, and ß-glucuronidase production. We aimed to investigate the role of these factors in the formation of gallstone in in vitro conditions at a supersaturated concentration of cholesterol. METHODOLOGY: To mimic bile in in vitro state, Brilliant Green Bile Broth (BGBB) media having a composition similar to human bile was used. Four different experimental sets were prepared, each having nine flasks with varying concentrations of cholesterol and CaCO3 (calcium carbonate). Test sets I, II, III, and IV were inoculated with Salmonella, Enterococcus, Helicobacter, and Neisseria respectively, which were isolated from gallstone itself. Out of these four bacteria, only Helicobacter did not possess slime activity. A control set was also established which was devoid of bacteria. The control also had nine flasks with different concentrations of cholesterol and CaCO3. All the sets were incubated in the incubator shaker at 37 °C and 80 revolution per minutes (RPM) for 20 days. RESULT: It was observed that the sets having bacteria had a less nucleation time as compared to the control (F = 5.274; p < 0.001). Solidification of gallstone was observed only in the set with bacteria having slime activity (sets I, II, and IV). CONCLUSION: The slime activity of bacteria leads to solidification of gallstones, whereas the other activities accelerate the nucleation of gallstone formation enhancing the severity of the disease.
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Bilis/metabolismo , Carbonato de Calcio/metabolismo , Colesterol/metabolismo , Cálculos Biliares/etiología , Cálculos Biliares/microbiología , Enterococcus , Cálculos Biliares/metabolismo , Glucuronidasa/metabolismo , Helicobacter , Técnicas In Vitro , Neisseria , Salmonella , Temperatura , Factores de Tiempo , Ureasa/metabolismoRESUMEN
BACKGROUND: Gallbladder stone is recently increased among the Iraqi society due to many risk factors such as bacterial infection and some HLA class II antigens. AIM(S): This study investigates the types of bacterial infection and HLA-DRB1 antigens' ratio that may be correlated with gallbladder stone formation. Setting and Design: The study included 45 patients and the same number of healthy individuals as a control group. Patients were with multiple gallstones. Gallstone bacterial culture was demonstrated to diagnose viable bacteria. HLA-DRB1 alleles' frequency was investigated using sequence-specific oligonucleotide probes (PCR-SSOP). RESULTS: Irrespective of gallstone type and size, different types of living viable bacteria were isolated from the cores of the studied gallstones in 80% of the studied cases versus 20% of sterile gallstones. Gram-negative bacteria cultures were the dominant (89.3%), including Escherichia coli, Klebsiella spp., Proteus spp., Acinetobacter spp., and Enterobacter spp. Mixed infection of Gram-positive and negative bacteria was noted: Escherichia coli and Enterococus spp. and the others of Escherichia coli and Acitobacter spp., and Klebsiella spp. and Pseudomonas spp. Gram-positive bacteria cultures were also detected at lower rate (10.7%) including Staphylococci spp. The frequency of HLA-DRB1*03:01, HLA-DRB1*4:03, HLA-DRB1*13:22, and HLA-DRB1*15:10 alleles was significantly elevated in patients compared to the healthy control group. CONCLUSION: Results ensured the viability of the bacteria isolated from the core of gallstones and showed positive correlation between gallbladder stone and different bacterial infection. In addition, HLA-DRB1 alleles were significantly high in patients compared to healthy control group suggesting them as risk factors (P < 0.05).
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Infecciones Bacterianas/complicaciones , Cálculos Biliares/inmunología , Cálculos Biliares/microbiología , Cadenas HLA-DRB1/genética , Adulto , Anciano , Anciano de 80 o más Años , Bacterias/clasificación , Bacterias/aislamiento & purificación , Infecciones Bacterianas/clasificación , Recuento de Colonia Microbiana , Femenino , Cálculos Biliares/genética , Frecuencia de los Genes , Humanos , Irak , Masculino , Persona de Mediana Edad , Factores de RiesgoRESUMEN
BACKGROUND: The gut microbiota participates in the metabolism of substances and energy, promotes the development and maturation of the immune system, forms the mucosal barrier, and protects the host from pathogen attacks. Although the pathogenesis of cholesterol gallstones is still not clear, studies have suggested that gut microbiota dysbiosis plays an important role in their formation. METHODS: Microbial DNA from faeces of normal control patients and those of patients with calculi was subjected to 16S rRNA gene sequencing to detect gene expression changes in intestinal microbes. ELISA kits were used to measure free bile acids, secondary bile acids and coprostanol according to the manufacturer's instructions. The relationship between flora and their metabolites was then analysed. RESULTS: In the gallstone group, the diversity of intestinal bacteria and the abundances of certain phylogroups were significantly decreased (p < 0.05), especially Firmicutes (p < 0.05), the largest phylum represented by the gut microbiota. This study found an increase in free bile acids (p < 0.001) and secondary bile acids (p < 0.01) in the enterohepatic circulation. Bile salt hydrolase activity was not related to the abundances of BSH-active bacteria. 7a-dehydroxylating gut bacteria were significantly increased (p < 0.01), whereas cholesterol-lowering bacteria were significantly reduced (p < 0.05). The Ruminococcus gnavus group could be used as a biomarker to distinguish the gallstone group from the control group. CONCLUSION: We conclude that intestinal flora imbalance affects bile acid and cholesterol metabolism and is associated with gallstone formation.
Asunto(s)
Ácidos y Sales Biliares/metabolismo , Cálculos Biliares/metabolismo , Cálculos Biliares/microbiología , Microbioma Gastrointestinal , Adulto , Bacterias/clasificación , Bacterias/genética , Colesterol/metabolismo , ADN Bacteriano/análisis , Disbiosis/microbiología , Circulación Enterohepática , Heces/microbiología , Femenino , Microbioma Gastrointestinal/genética , Expresión Génica , Humanos , Masculino , Persona de Mediana Edad , ARN Ribosómico 16S/genética , Análisis de Secuencia de ARNRESUMEN
BACKGROUND AND OBJECTIVE: The distal cholangiocarcinoma (dCCA) is associated with many factors: genes, environment, infection, etc. The current changes in biliary flora are thought to be involved in the formation of many gastrointestinal tract (GIT) diseases, like colon adenocarcinoma. Therefore we want to investigate whether the dCCA has a certain correlation with biliary microecology, and to detect specific strains. METHODS: A total of 68 adults were enrolled, of whom 8 with dCCA, 16 with recurrent choledocholithiasis, and 44 with the onset of common bile duct stones. Endoscopic Retrograde Cholangiopancretography (ERCP) was utilized to collect bile samples for DNA extraction and 16S rRNA gene sequencing, followed by analysis of bile microbiota composition. RESULTS: First, Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria are the most dominant phyla in the bile of patients with dCCA and the onset of common bile duct stoes. Secondly, compared with the onset of common bile duct stones patients, we got a significant increase in the phylum Gemmatimonadetes, Nitrospirae, Chloroflexi, Latescibacteria, and Planctomycetes in dCCA patients. Finally, at the genus level, we obtained sequencing results of 252 bacterial genera from patients with dCCA, recurrent choledocholithiasis, and the new onset of common bile duct stones, revealing heterogeneity among individuals. CONCLUSION: To the best of our knowledge, this is the first study of the dysbiosis of bile flora in patients with dCCA. This micro-ecological disorder may be a decisive factor in the formation of dCCA. At the same time, for the first time, this study provides a test chart of biliary microbial populations that may be associated with recurrent choledocholithiasis. The compositional changes of the core microbial group of the biliary tract have potentially important biological and medical significance for the microbiological biliary disorders of dCCA.
Asunto(s)
Neoplasias de los Conductos Biliares/microbiología , Sistema Biliar/microbiología , Colangiocarcinoma/microbiología , Cálculos Biliares/microbiología , Microbiota , Adulto , Anciano , Anciano de 80 o más Años , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Biodiversidad , China , Coledocolitiasis/microbiología , Femenino , Humanos , Masculino , Microbiota/genética , Persona de Mediana Edad , Recurrencia Local de Neoplasia , ARN Ribosómico 16SRESUMEN
Non-typhoidal Salmonella spp.are Gram-negative bacilli, which typically cause a clinical picture of gastroenteritis and, less commonly, patients may become a chronic carrier of the pathogen within their gallbladder. We describe a rare clinical presentation of a non-typhoidal Salmonella spp. infection as acute calculus cholecystitis in an adult patient. Salmonella enterica subsp. Salamae (ST P4271) was grown from cholecystostomy fluid, and the patient subsequently underwent a laparoscopic cholecystectomy that demonstrated a necrotic gallbladder fundus. We advise that microbiological sampling of bile is essential, especially in the context of foreign travel, to detect unusual pathogens as in this case or common pathogens that may have unusual antimicrobial resistance. Given the necrotic gallbladder as in this case, we also advise that early cholecystectomy should be strongly considered in these patients.
Asunto(s)
Colecistitis Aguda/microbiología , Cálculos Biliares/microbiología , Infecciones por Salmonella/microbiología , Salmonella enterica/aislamiento & purificación , Adulto , Antibacterianos/uso terapéutico , Colecistectomía Laparoscópica , Colecistitis Aguda/tratamiento farmacológico , Colecistitis Aguda/cirugía , Terapia Combinada , Cálculos Biliares/tratamiento farmacológico , Cálculos Biliares/cirugía , Humanos , MasculinoRESUMEN
Typhoid fever, a human-specific disease, is primarily caused by the pathogen Salmonella enterica serovar Typhi. It is estimated that 3 to 5% of people infected with typhoid fever become chronic carriers. Studies have demonstrated that a mechanism of chronic carriage involves biofilm formation on gallstone surfaces. In the course of a previous study using a chronic carriage mouse model, a Salmonella enterica serovar Typhimurium isolate was recovered from a mouse gallstone that exhibited a 2-fold increase in biofilm formation over the wild type. In order to identify the gene(s) responsible for the phenotype, the genomic sequences of this isolate and others were determined and compared. These sequences identified single nucleotide polymorphisms (SNPs) in 14 genes. Mutations in the most promising candidates, envZ and rcsB, were created, but neither showed increased biofilm-forming ability separately or in combination. The hyperbiofilm isolate did, however, present variations in cellular appendages observable using different techniques and a preferential binding to cholesterol. The isolate was also examined for systemic virulence and the ability to colonize the gallbladder/gallstones in a mouse model of chronic infection, demonstrating a systemic virulence defect and decreased gallbladder/gallstone colonization. Finally, to determine if the appearance of hyperbiofilm isolates could be replicated in vitro and if this was a common event, wild-type Salmonella spp. were grown long term in vitro under gallbladder-mimicking conditions, resulting in a high proportion of isolates that replicated the hyperbiofilm phenotype of the original isolate. Thus, Salmonella spp. acquire random mutations under the gallbladder/gallbladder-simulating conditions that may aid persistence but negatively affect systemic virulence.IMPORTANCE Chronic carriers are the main reservoirs for the spread of typhoid fever in regions of endemicity. Salmonella Typhi forms biofilms on gallstones in order to persist. A strain with enhanced biofilm-forming ability was recovered after a nine-month chronic-carriage mouse study. After sequencing this strain and recreating some of the mutations, we could not duplicate the phenotype. The isolate did show a difference in flagella, a preference to bind to cholesterol, and a systemic virulence defect. Finally, gallbladder conditions were simulated in vitro After 60 days, there was a 4.5-fold increase in hyperbiofilm isolates when a gallstone was present. These results indicate that Salmonella spp. can undergo genetic changes that improve persistence in gallbladder albeit at the cost of decreased virulence.