RESUMEN
A facultative anaerobic, catalase-negative, lactic acid-producing, non-endospore-forming, Gram-positive coccoid bacterium, strain MT/JULY 2010T, was isolated from the stomach of a Helicobacter pylori-positive Japanese female patient with pre-neoplastic gastric mucosa. This bacterium can form clusters, chains or pairs or occurs singly in liquid media. The growth ranges for temperature, pH and NaCl concentration were 20-45 °C, pH 5.0-7.0 and 1-4%, respectively. Cells exhibited an unusual tumbling motion when observed under the microscope using a hanging drop assay. Observation under an electron microscope showed some unidentified structures and numerous fimbriae-like appendages. The combined results of 16S rRNA gene sequence analysis and the overall genome-related index [average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH)] classified the bacterium as a novel species of the genus Streptococcus. It forms a monophyletic cluster with Streptococcus parasanguinis ATCC 15912T in the phylogenetic tree with 94.4% ANI. Furthermore, the conventional DNA-DNA hybridization assay and dDDH of the two strains identified their relatedness as 62.5% and 58.7% (both below the 70% threshold), respectively. The DNA G+C content, determined by nuclease P1-digested genomic DNA, was 42.6 mol%, while the digital G+C content calculated in silico was 41.8 mol%. Based on the phenotypic and genotypic results, strain MT/JULY 2010T represents a novel species within the genus. The name Streptococcus mobilis sp. nov. is proposed. The type strain is MT/JULY 2010T (=ATCC BAA-2258T=NBRC 107862T=NCBI 948105T=V10/022878T).
Asunto(s)
Infecciones por Helicobacter , Filogenia , Streptococcus , Humanos , ARN Ribosómico 16S/genética , Composición de Base , ADN Bacteriano/genética , Femenino , Técnicas de Tipificación Bacteriana , Hibridación de Ácido Nucleico , Análisis de Secuencia de ADN , Helicobacter pylori/aislamiento & purificación , Streptococcus/clasificación , Streptococcus/genética , Streptococcus/aislamiento & purificación , Infecciones por Helicobacter/microbiología , Japón , Estómago/microbiología , Mucosa Gástrica/microbiología , Ácidos Grasos/análisisRESUMEN
BACKGROUND AND OBJECTIVES: Chronic tobacco smoking contributes to oral health problems, such as periodontitis and tooth decay, which can result from smoking-altered oral microbiomes. The impact of chronic tobacco smoking on the oral microbiome remains not fully understood. METHODS: This was a cross-sectional study comparing the oral salivary microbiomes of 20 chronic tobacco smokers and 23 nonsmoking controls, all of whom were Chinese males, using microbial 16S rRNA sequencing. The duration of smoking, age, and information on gingivitis were collected and analyzed using the nonparametric Mann-Whitney test and a regression model. RESULTS: The most increased and decreased oral microbiomes in smokers versus controls were the genera Streptococcus and Neisseria, respectively. After adjusting for age, gingivitis, smoking duration, and FDR, only the abundance of the Pectinatus genus and Streptococcus australis species was significantly increased in smokers compared to controls. DISCUSSION AND CONCLUSIONS: This study reveals that long-term oral tobacco smoking is associated with the enrichment of some proinflammatory microbiomes, such as S. australis. SCIENTIFIC SIGNIFICANCE: Our study suggests that maintaining good oral hygiene, using oral probiotics that reduce proinflammatory microbiomes, or treating oral diseases may help prevent the pathogenesis of tobacco-enriched commensal pathobionts.
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Microbiota , Boca , Saliva , Fumar Tabaco , Humanos , Masculino , Microbiota/genética , Estudios Transversales , Adulto , Saliva/microbiología , Boca/microbiología , Persona de Mediana Edad , Streptococcus/aislamiento & purificación , Streptococcus/genética , ARN Ribosómico 16S , Neisseria/aislamiento & purificación , Neisseria/genética , Estudios de Casos y ControlesRESUMEN
Streptococcus dysgalactiae subsp. dysgalactiae is a pathogen that causes mastitis in dairy cows, resulting in significant economic losses in dairy production worldwide. This study aimed to identify the genetic determinants of S. dysgalactiae that are required for its growth in milk and adhesion to mammary epithelial cells. Transcriptome analysis revealed 936 genes that were significantly differentially expressed in milk compared with optimal medium, among which the gene with the greatest upregulation was dppA, which putatively encodes a lipoprotein tethered to the membrane with a lipid anchor. The dppA gene deletion significantly impaired the tolerance of lactoferrin, an antimicrobial protein in cow milk, and the ability to adhere to mammary epithelial cells, but did not affect growth in milk. Moreover, another lipoprotein gene, mtsA, which is involved in metal transport, was identified in S. dysgalactiae. Unlike dppA, the deletion of mtsA not only impacts lactoferrin tolerance and cell adhesion but also results in growth inhibition in milk. Further studies suggested that this effect was caused by a disturbed iron utilization. This study identified critical genetic determinants required for growth in milk and cell adhesion, which might provide novel insights into the mechanisms of S. dysgalactiae pathogenesis.
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Leche , Streptococcus , Animales , Leche/microbiología , Streptococcus/genética , Streptococcus/crecimiento & desarrollo , Streptococcus/metabolismo , Adhesión Bacteriana , Bovinos , Femenino , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Lactoferrina/metabolismo , Células Epiteliales/microbiología , Perfilación de la Expresión Génica , Lipoproteínas/genética , Lipoproteínas/metabolismo , Infecciones Estreptocócicas/microbiología , Infecciones Estreptocócicas/veterinaria , Hierro/metabolismo , Regulación Bacteriana de la Expresión Génica , Mastitis Bovina/microbiología , Eliminación de GenRESUMEN
Despite numerous studies investigating intratumoral microorganisms and their significant roles in cancer initiation, progression, and treatment efficacy, a systematic understanding of intratumoral microorganisms remains lacking. Herein, we conducted a study using 16S rDNA data on seven types of cancer, comprising a total of 783 samples. It's worth noting that Pseudomonas, Streptococcus, and Prevotella were found to be shared with the microbial communities of the seven cancers, suggesting that these may be associated with the occurrence and development of cancers. We anticipate establishing a foundation for related research and exploring potential methods for cancer treatment.
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Bacterias , Biodiversidad , Microbiota , Neoplasias , Humanos , Neoplasias/microbiología , ARN Ribosómico 16S/genética , ADN Ribosómico/genética , ADN Ribosómico/química , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , ADN Bacteriano/genética , Filogenia , Análisis de Secuencia de ADN , Pseudomonas/genética , Pseudomonas/aislamiento & purificación , Prevotella/genética , Prevotella/aislamiento & purificación , Streptococcus/genética , Streptococcus/aislamiento & purificaciónRESUMEN
Survival strategy of bacteria is expanded by extrachromosomal elements (ECEs). However, their genetic diversity and functional roles for adaptability are largely unknown. Here, we discover a novel family of intracellular ECEs using 56 saliva samples by developing an efficient microbial DNA extraction method coupled with long-read metagenomics assembly. Even though this ECE family was not hitherto identified, our global prevalence analysis using 476 salivary metagenomic datasets elucidates that these ECEs reside in 74% of the population. These ECEs, which we named, "Inocles", are giant plasmid-like circular genomic elements of 395 kb in length, including Streptococcus as a host bacterium. Inocles encode a series of genes that contribute to intracellular stress tolerance, such as oxidative stress and DNA damage, and cell wall biosynthesis and modification involved in the interactions with oral epithelial cells. Moreover, Inocles exhibit significant positive correlations with immune cells and proteins responding to microbial infection in peripheral blood. Intriguingly, we examine and find their marked reductions among 68 patients of head and neck cancers and colorectal cancers, suggesting its potential usage for a novel biomarker of gastrointestinal cancers. Our results suggest that Inocles potentially boost the adaptive capacity of host bacteria against various stressors in the oral environment.
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Bacterias , Microbiota , Boca , Humanos , Saliva/microbiología , Microbiota/genética , Metagenómica/métodos , Boca/microbiología , Femenino , Masculino , Neoplasias Colorrectales/microbiología , Neoplasias de Cabeza y Cuello/microbiología , Metagenoma , Adulto , ADN Bacteriano/genética , Bacterias/genética , Persona de Mediana Edad , Streptococcus/genéticaRESUMEN
Bacterial strains that are genetically engineered to constitutively produce fluorescent proteins have aided our study of bacterial physiology, biofilm formation, and interspecies interactions. Here, we report on the construction and utilization of new strains that produce the blue fluorescent protein mTagBFP2, the green fluorescent protein sfGFP, and the red fluorescent protein mScarlet-I3 in species Streptococcus gordonii, Streptococcus mutans, and Streptococcus sanguinis. Gene fragments, developed to contain the constitutive promoter Pveg, the fluorescent gene of interest, as well as aad9, providing resistance to the antibiotic spectinomycin, were inserted into selected open reading frames on the chromosome that were both transcriptionally silent and whose loss caused no measurable changes in fitness. All strains, except for sfGFP in S. sanguinis, were validated to produce a detectable and specific fluorescent signal. Individual stains, along with extracellular polymeric substances (EPS) within biofilms, were visualized and quantified through either widefield or super-resolution confocal microscopy approaches. Finally, to validate the ability to perform single-cell-level analysis using the strains, we imaged and analyzed a triculture mixed-species biofilm of S. gordonii, S. mutans, and S. sanguinis grown with and without the addition of human saliva. Quantification of the loss in membrane integrity using a SYTOX dye revealed that all strains had increased loss of membrane integrity with water or human saliva added to the growth media, but the proportion of the population stained by the SYTOX dye varied by species. In all, these fluorescent strains will be a valuable resource for the continued study of oral microbial ecology.IMPORTANCEStreptococci are among the earliest colonizers of the soft and hard tissues of the oral cavity and are contributors to the oral health status of the host, with involvement in dental caries, endodontic infections, periodontal disease, and the development of oral cancer. Strains genetically modified to produce fluorescent proteins that can be either visualized through microscopy imaging or quantified by their specific fluorescent intensity signal are critical tools toward the study of individual or mixed-species cultures. Our report here details the development and testing of several new strains of fluorescent oral streptococci that can be utilized in the study of microbial ecology, increasing both the availability of tools and documenting experimental approaches toward in vitro assay applications such as the study of intermicrobial interactions.
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Proteínas Luminiscentes , Boca , Streptococcus , Biopelículas/crecimiento & desarrollo , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Humanos , Streptococcus gordonii/genética , Streptococcus/genética , Streptococcus/metabolismo , Streptococcus/fisiología , Boca/microbiología , Streptococcus mutans/genética , Streptococcus mutans/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Proteína Fluorescente Roja , Streptococcus sanguis/genética , Microscopía ConfocalRESUMEN
Transformation is a mechanism of horizontal gene transfer widespread in bacteria. The first step in transformation-capture of exogenous DNA-is mediated by surface-exposed filaments belonging to the type 4 filament (T4F) superfamily. How these protein polymers, composed of major and minor pilin subunits, interact with DNA remains poorly understood. Here, we address this question for the Com pilus, a widespread T4F mediating DNA capture in competent monoderm species. Our functional analysis, performed in Streptococcus sanguinis, was guided by a complete structural model of the Com pilus. We show that the major pilin ComGC does not bind DNA. In contrast, a systematic mutational analysis of electropositive residues exposed at the filament surface in the four minor pilins (ComGD, ComGE, ComGF, and ComGG) reveals that the interface between ComGD and ComGF is important for DNA capture. Sequential mutations in these two interacting subunits lead to complete abolition of transformation, without affecting piliation. We further demonstrate the physical interaction between ComGD and ComGF using disulfide crosslinking, upon mutagenesis of two strategically positioned residues into cysteines. A structural model of the Com pilus tip interacting with DNA recapitulates all these findings and highlights a novel mode of DNA-binding, conserved in hundreds of monoderm species. IMPORTANCE: Bacteria are capable of evolving and diversifying very rapidly by acquiring new genetic material via horizontal gene transfer (HGT). Transformation is a widespread mechanism of HGT, which results from the capture of extracellular DNA by surface-exposed pili belonging to the type 4 filament (T4F) superfamily. How T4F-composed of major and minor pilins-interact with DNA remains poorly understood, especially in monoderm species that use a unique T4F for DNA capture, known as Com pilus or T4dP. The significance of this work is in characterizing a novel mode of DNA-binding by showing that the interface between two minor pilins, part of a tip-located complex of four pilins-found in different T4F-has been functionalized in monoderms to capture DNA. This is an evolutionary mechanism promoting the exceptional functional versatility of T4F.
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ADN Bacteriano , Proteínas Fimbrias , Fimbrias Bacterianas , Streptococcus , Transformación Bacteriana , Fimbrias Bacterianas/metabolismo , Fimbrias Bacterianas/genética , Fimbrias Bacterianas/química , Proteínas Fimbrias/metabolismo , Proteínas Fimbrias/genética , Proteínas Fimbrias/química , ADN Bacteriano/metabolismo , ADN Bacteriano/genética , Modelos Moleculares , Streptococcus/genética , Streptococcus/metabolismo , Unión Proteica , Transferencia de Gen HorizontalRESUMEN
BACKGROUND: Cystic fibrosis (CF) lung disease begins early, and prophylactic antibiotics have been used to prevent Staphylococcus aureus infection. This study examined the lung microbiome in two countries with differing antibiotic practices and its relationship to lung function in young children with CF. METHODS: A binational, longitudinal, observational study was performed to define the lower airway microbiome in infants with CF. 16S rRNA sequencing was performed using lavage fluid to characterize the lung microbiota in 45 infants with and without prophylactic antibiotic therapy at an average age of approximately 3 months and 14 months. The association between pulmonary function, bacterial community diversities, and taxa was assessed. RESULTS: Expected CF bacterial genera and non-traditional bacteria, such as Streptococcus, were identified as core taxa. Microbial community shifts were observed in infants who received antibiotic prophylaxis, with lower alpha diversity (ANOVA, P < 0.05) and a higher proportion of Streptococcus at the first visit. Beta diversity (FEV0.5z; MiRKAT, P < 0.05) and Streptococcus were associated with FEV0.5z (LASSO and linear regression, ß < 0). Functional annotation suggested that alteration of lung microbiota may be linked to antimicrobial resistance. CONCLUSIONS: Lung microbial diversity in infants with CF varied between the two countries, particularly during early infancy. A shift in the lung microbiome toward a higher relative abundance of Streptococcus was associated with reduced pulmonary function.
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Antibacterianos , Fibrosis Quística , Pulmón , Microbiota , Humanos , Fibrosis Quística/microbiología , Fibrosis Quística/fisiopatología , Fibrosis Quística/tratamiento farmacológico , Pulmón/microbiología , Pulmón/fisiopatología , Microbiota/efectos de los fármacos , Microbiota/genética , Lactante , Antibacterianos/uso terapéutico , Femenino , Masculino , Estudios Longitudinales , ARN Ribosómico 16S/genética , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Bacterias/efectos de los fármacos , Streptococcus/aislamiento & purificación , Streptococcus/genética , Profilaxis Antibiótica , Líquido del Lavado Bronquioalveolar/microbiología , Pruebas de Función RespiratoriaRESUMEN
NrdR is a universal transcriptional repressor of bacterial genes coding for ribonucleotide reductases (RNRs), essential enzymes that provide DNA building blocks in all living cells. Despite its bacterial prevalence, the NrdR mechanism has been scarcely studied. We report the biochemical, biophysical, and bioinformatical characterization of NrdR and its binding sites from two major bacterial pathogens of the phylum Bacillota Listeria monocytogenes and Streptococcus pneumoniae. NrdR consists of a Zn-ribbon domain followed by an ATP-cone domain. We show that it forms tetramers that bind to DNA when loaded with ATP and dATP, but if loaded with only ATP, NrdR forms various oligomeric complexes unable to bind DNA. The DNA-binding site in L. monocytogenes is a pair of NrdR boxes separated by 15-16 bp, whereas in S. pneumoniae, the NrdR boxes are separated by unusually long spacers of 25-26 bp. This observation triggered a comprehensive binding study of four NrdRs from L. monocytogenes, S. pneumoniae, Escherichia coli, and Streptomyces coelicolor to a series of dsDNA fragments where the NrdR boxes were separated by 12-27 bp. The in vitro results were confirmed in vivo in E. coli and revealed that NrdR binds most efficiently when there is an integer number of DNA turns between the center of the two NrdR boxes. The study facilitates the prediction of NrdR binding sites in bacterial genomes and suggests that the NrdR mechanism is conserved throughout the bacterial domain. It sheds light on RNR regulation in Listeria and Streptococcus, and since NrdR does not occur in eukaryotes, opens a way to the development of novel antibiotics.
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Proteínas Bacterianas , Listeria monocytogenes , Listeria , Proteínas Represoras , Ribonucleótido Reductasas , Streptococcus pneumoniae , Streptococcus , Ribonucleótido Reductasas/metabolismo , Ribonucleótido Reductasas/genética , Listeria monocytogenes/genética , Listeria monocytogenes/metabolismo , Listeria monocytogenes/enzimología , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/metabolismo , Streptococcus pneumoniae/enzimología , Sitios de Unión , ADN Bacteriano/metabolismo , Proteínas Represoras/metabolismo , Proteínas Represoras/genética , Unión Proteica , Regulación Bacteriana de la Expresión Génica , Proteínas de Unión al ADN/metabolismo , Proteínas de Unión al ADN/genética , Listeria/genética , Listeria/metabolismo , Streptococcus/genética , Streptococcus/metabolismo , Adenosina Trifosfato/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismoRESUMEN
OBJECTIVES: An increase in Streptococcus dysgalactiae subsp. equisimilis (SDSE) infections has been documented worldwide. This study aims to analyze invasive disease caused by SDSE (iSDSE) in adults over an 11-year period in Spain. METHODS: We conducted a retrospective, laboratory-based study of iSDSE detected at Hospital Universitari de Bellvitge (HUB) from 2012 to 2022 (n = 89) and isolates collected in three Spanish hospitals in 2018 (n = 22). Clinical data from HUB were collected. Isolates were tested for antimicrobial susceptibility (European Committee on Antimicrobial Susceptibility Testing 2023), subjected to whole genome sequencing and analyzed for mobile genetic elements (MGEs). A mouse model was used to analyze virulence. RESULTS: iSDSE episodes at HUB occurred predominantly in older patients with comorbidities (particularly, diabetes, chronic heart disease, and malignancies). Whole genome sequencing revealed a high genetic diversity, with the most common lineages being CC15, CC17, and CC20. Various virulence factors, including the superantigen speG, were identified. Macrolides, lincosamides, and tetracyclines exhibited the highest resistance rates (>27%) and changed over time, linked to multiple MGEs. The mouse model highlighted the virulence of the CC20-stG62647 lineage, but these results were discordant with clinical data. CONCLUSION: iSDSE incidence is increasing and associated with older patients with comorbidities. Genetically, SDSE is diverse with a high capacity to integrate MGEs carrying resistance determinants. Mouse model studies showed the enhanced virulence of the CC20-stG62647 lineage. These findings underscore the need for ongoing surveillance of this emerging pathogen.
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Infecciones Estreptocócicas , Streptococcus , España/epidemiología , Infecciones Estreptocócicas/epidemiología , Infecciones Estreptocócicas/microbiología , Infecciones Estreptocócicas/tratamiento farmacológico , Humanos , Streptococcus/genética , Streptococcus/patogenicidad , Streptococcus/efectos de los fármacos , Streptococcus/aislamiento & purificación , Streptococcus/clasificación , Animales , Femenino , Ratones , Estudios Retrospectivos , Masculino , Anciano , Antibacterianos/farmacología , Persona de Mediana Edad , Factores de Virulencia/genética , Pruebas de Sensibilidad Microbiana , Virulencia , Adulto , Secuenciación Completa del Genoma , Genoma Bacteriano , Anciano de 80 o más AñosRESUMEN
Introduction: Changes in the human gut microbiome have been linked to various chronic diseases, including chronic obstructive pulmonary disease (COPD). While substantial knowledge is available on the genomic features of fecal communities, little is known about the microbiome's transcriptional activity. Here, we analyzed the metatranscriptomic (MTR) abundance of MetaCyc pathways, SuperPathways, and protein domain families (PFAM) represented by the gut microbiome in a cohort of non-small cell lung cancer (NSCLC) patients with- or without COPD comorbidity. Methods: Fecal samples of 40 NSCLC patients with- or without COPD comorbidity were collected at the time of diagnosis. Data was preprocessed using the Metaphlan3/Humann3 pipeline and BioCyc© to identify metabolic SuperPathways. LEfSe analysis was conducted on Pathway- and PFAM abundance data to determine COPD- and non-COPD-related clusters. Results: Key genera Streptococcus, Escherichia, Gemella, and Lactobacillus were significantly more active transcriptionally compared to their metagenomic presence. LEfSe analysis identified 11 MetaCyc pathways that were significantly overrepresented in patients with- and without COPD comorbidity. According to Spearman's rank correlation, Smoking PY showed a significant negative correlation with Glycolysis IV, Purine Ribonucleoside Degradation and Glycogen Biosynthesis I, and a significant positive correlation with Superpathway of Ac-CoA Biosynthesis and Glyoxylate cycle, whereas forced expiratory volume in the first second (FEV1) showed a significant negative correlation with Glycolysis IV and a significant positive correlation with Glycogen Biosynthesis I. Furthermore, COPD patients showed a significantly increased MTR abundance in ~60% of SuperPathways, indicating a universally increased MTR activity in this condition. FEV1 showed a significant correlation with SuperPathways Carbohydrate degradation, Glycan biosynthesis, and Glycolysis. Taxonomic analysis suggested a more prominent MTR activity from multiple Streptococcus species, Enterococcus (E.) faecalis, E. faecium and Escherichia (E.) coli than expected from their metagenomic abundance. Multiple protein domain families (PFAMs) were identified as more associated with COPD, E. faecium, E.coli, and Streptococcus salivarius, contributing the most to these PFAMs. Conclusion: Metatranscriptome analysis identified COPD-related subsets of lung cancer with potential therapeutic relevance.
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Heces , Microbioma Gastrointestinal , Neoplasias Pulmonares , Redes y Vías Metabólicas , Enfermedad Pulmonar Obstructiva Crónica , Humanos , Enfermedad Pulmonar Obstructiva Crónica/microbiología , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Microbioma Gastrointestinal/genética , Redes y Vías Metabólicas/genética , Neoplasias Pulmonares/microbiología , Neoplasias Pulmonares/genética , Masculino , Heces/microbiología , Femenino , Anciano , Persona de Mediana Edad , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/microbiología , Transcriptoma , Metagenómica/métodos , Bacterias/genética , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacterias/metabolismo , Streptococcus/genética , Streptococcus/aislamiento & purificación , Streptococcus/metabolismoRESUMEN
This study aimed to screen the bioactive components in Streptococcus equinus WC1 (SE-WC1) and Limosilactobacillus reuteri GM4 (LR-GM4) and estimate the therapeutic role in Ehrlich solid tumors (EST) mice model. Forty-four male albino EST mice were assigned into 7 groups and treated daily for 2 weeks, including the EST group, the EST mice that received SE-WC1 at a low or a high dose (0.5 ml *106 or 0.5 ml *108 cfu), the EST mice that received LR-GM4 at the low or the high dose (0.5 ml *106 or 0.5 ml *108 cfu), and the EST mice that received SE-WC1 plus LR-GM4 at the low or the high dose. Tumors were harvested, weighed, examined, and used for the determination of apoptosis-related gene expression. Samples of the intestine, liver, and kidney were gathered for histological examination. The GC-MS identified 24 and 36 bioactive compounds in SE-WC1 and LR-GM4, respectively. The main compound in SE-WC1 was lupeol; however, the main compound in LR-GM4 was retinaldehyde. EST mice showed disturbances in Bcl-2, Bax, and p53 mRNA expression along with histological changes in the intestine, liver, and kidney. Administration of both bacterial strains reduced the tumor weight, alleviated the disturbances in the gene expression, and improved the histological structure of the intestine, liver, and kidney in a dose-dependent. Moreover, LR-GM4 was more effective than SE-WC1 due to its higher content of bioactive compounds. It could be concluded that these strains of probiotics are promising for the treatment of solid tumors.
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Carcinoma de Ehrlich , Limosilactobacillus reuteri , Probióticos , Animales , Probióticos/administración & dosificación , Ratones , Masculino , Carcinoma de Ehrlich/metabolismo , Carcinoma de Ehrlich/terapia , Limosilactobacillus reuteri/metabolismo , Streptococcus/metabolismo , Streptococcus/genética , Metabolismo Secundario , Apoptosis/efectos de los fármacos , Modelos Animales de Enfermedad , Hígado/metabolismoRESUMEN
Four Gram-stain-positive bacterial strains (designated 475-2T, 46-6BT, 778-2T and A810-3), isolated from traditional Chinese pickle, were characterized using a polyphasic taxonomic approach. Strain 475-2T was most closely related to the type strain of Lapidilactobacillus achengensis, having 99.9% 16S rRNA gene sequence similarity, 94.1-95.1% average nucleotide identity (ANI) and 57.6% digital DNA-DNA hybridization (dDDH) values. Strain 46-6BT was most closely related to the type strain of Secundilactobacillus similis, having 99.8% 16S rRNA gene sequence similarity, 94.3-94.9% ANI and 58.9-59.2% dDDH values. Strains 778-2T and A810-3 were phylogenetically related to the type strains of Streptococcus salivarius, Streptococcus thermophilus and Streptococcus vestibularis, having 99.7-99.9% 16S rRNA gene sequence similarities, 89.1-94.4% ANI and 39.0-55.5% dDDH values. Based upon the data obtained in the present study, three novel species, Lapidilactobacillus salsurivasis sp. nov., Secundilactobacillus muriivasis sp. nov. and Streptococcus parasalivarius sp. nov., are proposed and the type strains are 475-2T (= JCM 36613T = CCTCC AB 2023258T = LMG 33412T), 46-6BT (= JCM 36612T = CCTCC AB 2023259T = LMG 33411T) and 778-2T (= JCM 36614T = CCTCC AB 2023257T = LMG 33413T), respectively.
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ADN Bacteriano , Filogenia , ARN Ribosómico 16S , Streptococcus , ARN Ribosómico 16S/genética , ADN Bacteriano/genética , Streptococcus/genética , Streptococcus/clasificación , Streptococcus/aislamiento & purificación , Técnicas de Tipificación Bacteriana , China , Hibridación de Ácido Nucleico , Alimentos Fermentados/microbiología , Análisis de Secuencia de ADN , Composición de Base , Microbiología de Alimentos , Ácidos Grasos/análisisRESUMEN
AIMS: This study evaluates the antibacterial characteristics and mechanisms of combined tea polyphenols (TPs), Nisin, and ε-polylysine (PL) against Streptococcus canis, Streptococcus minor, Streptococcus mutans, and Actinomyces oris, common zoonotic pathogens in companion animals. METHODS AND RESULTS: Pathogenic strains were isolated from feline oral cavities and assessed using minimum inhibitory concentration (MIC) tests, inhibition zone assays, growth kinetics, and biofilm inhibition studies. Among single agents, PL exhibited the lowest MIC values against all four pathogens. TP showed significant resistance against S. minor, and Nisin against S. mutans. The combination treatment (Comb) of TP, Nisin, and PL in a ratio of 13:5:1 demonstrated broad-spectrum antibacterial activity, maintaining low MIC values, forming large inhibition zones, prolonging the bacterial lag phase, reducing growth rates, and inhibiting biofilm formation. RNA sequencing and metabolomic analysis indicated that TP, Nisin, and PL inhibited various membrane-bound carbohydrate-specific transferases through the phosphoenolpyruvate-dependent phosphotransferase system in S. canis, disrupting carbohydrate uptake. They also downregulated glycolysis and the citric acid cycle, inhibiting cellular energy metabolism. Additionally, they modulated the activities of peptidoglycan glycosyltransferases and d-alanyl-d-alanine carboxypeptidase, interfering with peptidoglycan cross-linking and bacterial cell wall stability. CONCLUSIONS: The Comb therapy significantly enhances antibacterial efficacy by targeting multiple bacterial pathways, offering potential applications in food and pharmaceutical antimicrobials.
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Antibacterianos , Biopelículas , Pruebas de Sensibilidad Microbiana , Nisina , Polilisina , Polifenoles , Té , Animales , Nisina/farmacología , Antibacterianos/farmacología , Polilisina/farmacología , Polifenoles/farmacología , Gatos , Té/química , Biopelículas/efectos de los fármacos , Streptococcus/efectos de los fármacos , Streptococcus/genética , Transcriptoma , Boca/microbiología , MetabolómicaRESUMEN
OBJECTIVE: Research has previously established connections between the intestinal microbiome and the progression of some cancers. However, there is a noticeable gap in the literature in regard to using Mendelian randomisation (MR) to delve into potential causal relationships between the gut microbiota (GM) and basal cell carcinoma (BCC). Therefore, the purpose of our study was to use MR to explore the causal relationship between four kinds of GM (Bacteroides, Streptococcus, Proteobacteria and Lachnospiraceae) and BCC. METHODS: We used genome-wide association study (GWAS) data and MR to explore the causal relationship between four kinds of GM and BCC. This study primarily employed the random effect inverse variance weighted (IVW) model for analysis, as complemented by additional methods including the simple mode, weighted median, weighted mode and MRâEgger methods. We used heterogeneity and horizontal multiplicity to judge the reliability of each analysis. MR-PRESSO was mainly used to detect and correct outliers. RESULTS: The random-effects IVW results showed that Bacteroides (OR = 0.936, 95% CI = 0.787-1.113, p = 0.455), Streptococcus (OR = 0.974, 95% CI = 0.875-1.083, p = 0.629), Proteobacteria (OR = 1.113, 95% CI = 0.977-1.267, p = 0.106) and Lachnospiraceae (OR = 1.027, 95% CI = 0.899-1.173, p = 0.688) had no genetic causal relationship with BCC. All analyses revealed no horizontal pleiotropy, heterogeneity or outliers. CONCLUSION: We found that Bacteroides, Streptococcus, Proteobacteria and Lachnospiraceae do not increase the incidence of BCC at the genetic level, which provides new insight for the study of GM and BCC.
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Carcinoma Basocelular , Microbioma Gastrointestinal , Estudio de Asociación del Genoma Completo , Análisis de la Aleatorización Mendeliana , Neoplasias Cutáneas , Humanos , Carcinoma Basocelular/genética , Carcinoma Basocelular/microbiología , Microbioma Gastrointestinal/genética , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/microbiología , Streptococcus/genética , Proteobacteria/genética , Bacteroides/genética , Predisposición Genética a la Enfermedad/genética , Polimorfismo de Nucleótido SimpleRESUMEN
There is a growing interest in the use of probiotic bacteria as biosensors for the detection of disease. However, there is a lack of bacterial receptors developed for specific disease biomarkers. Here, we have investigated the use of the peptide-regulated transcription factor ComR from Streptococcus spp. for specific peptide biomarker detection. ComR exhibits a number of attractive features that are potentially exploitable to create a biomolecular switch for engineered biosensor circuitry within the probiotic organism Lactiplantibacillus plantarum WCFS1. Through iterative design-build-test cycles, we developed a genomically integrated, ComR-based biosensor circuit that allowed WCFS1 to detect low nanomolar concentrations of ComR's cognate peptide XIP. By screening a library of ComR proteins with mutant residues substituted at the K100 position, we identified mutations that increased the specificity of ComR toward an amidated version of its cognate peptide, demonstrating the potential for ComR to detect this important class of biomarker.IMPORTANCEUsing bacteria to detect disease is an exciting possibility under active study. Detecting extracellular peptides with specific amino acid sequences would be particularly useful as these are important markers of health and disease (biomarkers). In this work, we show that a probiotic bacteria (Lactiplantibacillus plantarum) can be genetically engineered to detect specific extracellular peptides using the protein ComR from Streptococcus bacteria. In its natural form, ComR allowed the probiotic bacteria to detect a specific peptide, XIP. We then modified XIP to be more like the peptide biomarkers found in humans and engineered ComR so that it activated with this modified XIP and not the original XIP. This newly engineered ComR also worked in the probiotic bacteria, as expected. This suggests that with additional engineering, ComR might be able to activate with human peptide biomarkers and be used by genetically engineered probiotic bacteria to better detect disease.
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Proteínas Bacterianas , Péptidos , Factores de Transcripción , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Péptidos/metabolismo , Péptidos/genética , Probióticos/metabolismo , Mutación , Técnicas Biosensibles , Lactiplantibacillus plantarum/genética , Lactiplantibacillus plantarum/metabolismo , Regulación Bacteriana de la Expresión Génica , Streptococcus/genética , Streptococcus/metabolismoRESUMEN
MISTRG is an immunodeficient mouse strain that expresses multiple human cytokines that support hematopoietic stem cell maintenance and myelopoiesis. While establishing a breeding colony of MISTRG mice in a dedicated barrier room, 6 cases of death or disease occurred in pregnant or postpartum mice. Clinically, this manifested as hunched posture, dyspnea, and 1 case of emaciation with ataxia. Pathologic analysis of 7 mice revealed multisystemic necrosuppurative inflammation variably affecting the uterus and placenta, joints, meninges, inner and middle ears, kidneys, and small intestine. Bacteria cultured from the blood of septic mice were identified with 89% probability by the Vitek 2 identification system as Streptococcus sanguinus with atypical biochemical parameters; the API 20E/NE system fully differentiated the isolates as a novel Streptococcus species. MALDI Biotyper-based mass spectrometry also indicated that the phenotype represented a novel Streptococcus spp. Sequencing revealed that the full-length 16S rRNA gene identity was below 97% with known Streptococcus species, including the 2 closest species Streptococcus acidominimus and Streptococcus azizii. We propose the name Streptococcus murisepticum spp. nov to our novel isolates. All male mice in this colony remained healthy despite their association with diseased female mice. Overall, 19% of the colony carried the novel Streptococcus in their oral cavity, but it could not be detected in feces. The organism was sensitive to amoxicillin, which was administered via drinking water throughout pregnancy and weaning to establish a colony of pathogen-negative future breeders. The colony remained disease-free and culture-negative for Streptococcus murisepticum spp. nov after treatment with amoxicillin. We suspect that oral colonization of MISTRG mice with the novel Streptococcus species and its associated unique pathology in periparturient mice is potentially the principal cause of loss of this strain at several institutions. Therefore, screening the oral cavity for α-hemolytic streptococci followed by targeted antibiotic treatment may be necessary when establishing MISTRG and allied immunodeficient mouse strains.
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Infecciones Estreptocócicas , Embarazo , Masculino , Femenino , Humanos , Animales , Ratones , Infecciones Estreptocócicas/diagnóstico , ARN Ribosómico 16S/genética , Streptococcus/genética , Amoxicilina , BocaRESUMEN
Human oral microbial communities are diverse, with implications for oral and systemic health. Oral microbial communities change over time; thus, it is important to understand how healthy versus dysbiotic oral microbiomes differ, especially within and between families. There is also a need to understand how the oral microbiome composition is changed within an individual including by factors such as environmental tobacco smoke (ETS) exposure, metabolic regulation, inflammation, and antioxidant potential. Using archived saliva samples collected from caregivers and children during a 90-month follow-up assessment in a longitudinal study of child development in the context of rural poverty, we used 16S rRNA gene sequencing to determine the salivary microbiome. A total of 724 saliva samples were available, 448 of which were from caregiver/child dyads, an additional 70 from children and 206 from adults. We compared children's and caregivers' oral microbiomes, performed "stomatotype" analyses, and examined microbial relations with concentrations of salivary markers associated with ETS exposure, metabolic regulation, inflammation, and antioxidant potential (i.e., salivary cotinine, adiponectin, C-reactive protein, and uric acid) assayed from the same biospecimens. Our results indicate that children and caregivers share much of their oral microbiome diversity, but there are distinct differences. Microbiomes from intrafamily individuals are more similar than microbiomes from nonfamily individuals, with child/caregiver dyad explaining 52% of overall microbial variation. Notably, children harbor fewer potential pathogens than caregivers, and participants' microbiomes clustered into two groups, with major differences being driven by Streptococcus spp. Differences in salivary microbiome composition associated with ETS exposure, and taxa associated with salivary analytes representing potential associations between antioxidant potential, metabolic regulation, and the oral microbiome. IMPORTANCE The human oral cavity is a multi-environment habitat that harbors a diversity of microorganisms. This oral microbiome is often transmitted between cohabitating individuals, which may associate oral and systemic health within family members. Furthermore, family social ecology plays a significant role in childhood development, which may be associated with lifelong health outcomes. In this study, we collected saliva from children and their caregivers and used 16S rRNA gene sequencing to characterize their oral microbiomes. We also analyzed salivary biomeasures of environmental tobacco smoke exposure, metabolic regulation, inflammation, and antioxidant potential. We show there are differences in individuals' oral microbiomes mainly due to Streptococcus spp. that family members share much of their microbes, and several bacterial taxa associate with the selected salivary biomeasures. Our results suggest there are large-scale oral microbiome patterns, and there are likely relationships between oral microbiomes and the social ecology of families.
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Microbiota , Contaminación por Humo de Tabaco , Adulto , Humanos , Niño , Contaminación por Humo de Tabaco/efectos adversos , Cuidadores , Estudios Longitudinales , ARN Ribosómico 16S/genética , Antioxidantes , Microbiota/genética , Streptococcus/genética , InflamaciónRESUMEN
The aim of this study was to evaluate the effect of Lactobacillus delbrueckii subsp. lactis (L.del) on vaginal microbiota (VM) dysbiosis and vaginal radiation injury in gynecologic cancer patients. The inhibitory effects of L.del on cervical cancer cells were also studied in vitro. Gynecologic cancer patients receiving radiotherapy were randomized into control and L.del intervention groups. The control group received radiotherapy, while the intervention group received radiotherapy and L.del intervention (1 capsule/day placed into the deep vagina from the first day of radiotherapy until the end of treatment). Vaginal swab samples were collected on the first day pre-treatment and the last day post-treatment. DNA from 54 patients was extracted and assessed by the 16S rRNA sequencing method. Radiotherapy resulted in vaginal microbiome dysbiosis characterized by increased phylogenetic diversity and increased abundance of Brevundimonas, Streptococcus and Prevotella, but a decreased abundance of Lactobacillus. Level 2 vaginal radiation injury was positively associated with the abundance of Brevundimonas and gram-negative non-fermenting bacteria. Administration of L.del attenuated the reduction of Lactobacillus while also inhibiting the abundance of Streptococcus and Prevotella, thereby ameliorating radiotherapy-related vaginal microbiota dysbiosis. CLD inhibited the in vitro proliferation of SiHa cells by altering the expression of BCL2, HPV16-E6, HPV16-E7, IL6, MAP7, BAX, Caspase-3, Caspase-9 and LTF. In conclusion, L. del application can alleviate radiation-induced vaginal dysbiosis and restore Lactobacillus dominance of the vaginal microbiome. Moreover, CLD was found to inhibit cell growth and promote the apoptosis of SiHa cells in vitro. The registration number for this clinical trial is ChiCTR1900021784.
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Disbiosis , Neoplasias , Femenino , Humanos , Disbiosis/microbiología , ARN Ribosómico 16S/genética , Filogenia , Vagina/microbiología , Lactobacillus/genética , Streptococcus/genéticaRESUMEN
The ability to take up and incorporate foreign DNA via natural transformation is a well-known characteristic of some species of Streptococcus, and is a mechanism that rapidly allows for the acquisition of antibacterial resistance. Here, we describe that the understudied species Streptococcus ferus is also capable of natural transformation and uses a system analogous to that identified in Streptococcus mutans. S. mutans natural transformation is under the control of the alternative sigma factor sigX (also known as comX), whose expression is induced by two types of peptide signals: CSP (competence stimulating peptide, encoded by comC) and XIP (sigX-inducing peptide, encoded by comS). These systems induce competence via either the two-component signal-transduction system ComDE or the RRNPP transcriptional regulator ComR, respectively. Protein and nucleotide homology searches identified putative orthologs of comRS and sigX in S. ferus, but not homologs of S. mutans blpRH (also known as comDE). We demonstrate that natural transformation in S. ferus is induced by a small, double-tryptophan containing sigX-inducing peptide (XIP), akin to that of S. mutans, and requires the presence of the comR and sigX orthologs for efficient transformation. Additionally, we find that natural transformation is induced in S. ferus by both the native XIP and the XIP variant of S. mutans, implying that cross talk between the two species is possible. This process has been harnessed to construct gene deletions in S. ferus and provides a method to genetically manipulate this understudied species. IMPORTANCE Natural transformation is the process by which bacteria take up DNA and allows for acquisition of new genetic traits, including those involved in antibiotic resistance. This study demonstrates that the understudied species Streptococcus ferus is capable of natural transformation using a peptide-pheromone system like that previously identified in Streptococcus mutans and provides a framework for future studies concerning this organism.