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Quantitative investigation of the bacterial content of periodontal abscess samples by real-time PCR.Journal of Microbiological Methods Oct 2023Periodontal abscesses, which are part of the acute periodontal disease group characterized by the destruction of periodontal tissue with deep periodontal pockets,...
OBJECTIVES
Periodontal abscesses, which are part of the acute periodontal disease group characterized by the destruction of periodontal tissue with deep periodontal pockets, bleeding on probing, suppuration, and localized pus accumulation, cause rapid destruction of tooth-supporting tissues. This study aimed to evaluate the microbial content of periodontal abscesses by specific and culture-independent qPCR.
METHODS
This study was conducted on 30 volunteers diagnosed with periodontal abscesses and presenting with complaints of localized pain, swelling, and tenderness in the gingiva. Genomic DNA was isolated from the samples taken. Escherichia coli bacteria were used for the standard curve created to calculate the prevalence of target bacteria in the total bacterial load. 16S rRNA Universal primers were used to assess the total bacterial load and prevalence. Bacterial counts were analyzed with Spearman's rank correlation coefficients (ρ) matrix.
RESULTS
From the analysis of Real-Time PCR, Porphyromonas gingivalis (30, 100%), Prevotella intermedia (30, 100%), and Fusobacterium nucleatum (30, 100%) were detected in all samples. Campylobacter rectus (29, 96.6%), Porphyromonas endodontalis (29, 96.6%), Tannerella forsythia (28, 93.3%), Filifactor alocis (28, 93.3%), and Actinomyces naeslundii (28, 93.3%) were also frequently detected.
CONCLUSIONS
Periodontal abscesses were found to be polymicrobial, and not only periodontal pathogens appeared to be associated with the development of periodontal abscesses. The presence, prevalence, and number of Porphyromonas endodontalis and Propionibacterium acnes in the contents of periodontal abscesses were determined for the first time in our study. Further studies are needed to better understand the roles of bacteria in periodontal disease, including abscesses.
PubMed: 37742798
DOI: 10.1016/j.mimet.2023.106826 -
Scientific Reports Sep 2023The bacteriophage CAP 10-3 forming plaques against Cutibacterium acnes which causes skin acne was previously isolated from human skin acne lesion. Incomplete whole...
The bacteriophage CAP 10-3 forming plaques against Cutibacterium acnes which causes skin acne was previously isolated from human skin acne lesion. Incomplete whole genome sequence (WGS) of the bacteriophage CAP 10-3 was obtained and it had 29,643 bp long nucleotide with 53.86% GC content. The sequence was similar to C. acnes phage PAP 1-1 with a nucleotide sequence identity of 89.63% and the bacteriophage belonged to Pahexavirus. Bioinformatic analysis of the WGS predicted 147 ORFs and functions of 40 CDSs were identified. The predicted endolysin gene of bacteriophage CAP 10-3 was 858 bp long which was deduced as 285 amino acids (~ 31 kDa). The protein had the highest similarity with amino acid sequence of the endolysin from Propionibacterium phage PHL071N05 with 97.20% identity. The CAP 10-3 endolysin gene was amplified by PCR with primer pairs based on the gene sequence, cloned into an expression vector pET-15b and transformed into Escherichia coli BL21(DE3) strain. The predicted protein band (~ 33 kDa) for the recombinant endolysin was detected in an SDS-PAGE gel and western blot assay. The concentrated supernatant of cell lysate from E. coli BL21(DE3) (pET-15b_CAP10-3 end) and a partially purified recombinant CAP 10-3 endolysin showed antibacterial activity against C. acnes KCTC 3314 in a dose-dependent manner. In conclusion, the recombinant CAP 10-3 endolysin was successfully produced in E. coli strain and it can be considered as a therapeutic agent candidate for treatment of human skin acne.
Topics: Humans; Bacteriophages; Escherichia coli; Endopeptidases; Acne Vulgaris
PubMed: 37777575
DOI: 10.1038/s41598-023-43559-z -
Animal Health Research Reviews Dec 2023Animal husbandry is increasingly under pressure to meet world food demand. Thus, strategies are sought to ensure this productivity increment. The objective of this... (Review)
Review
Animal husbandry is increasingly under pressure to meet world food demand. Thus, strategies are sought to ensure this productivity increment. The objective of this review was to gather advances in the use of bacterial probiotics in animal production. Lactobacilli correspond to the most used bacterial group, with several beneficial effects already reported and described, as well as the and genera - being the latter expressively used in aquaculture. Research on the genus is mostly focused on human health, which demonstrates great effects on blood biochemical parameters. Such results sustain the possibility of expanding its use in veterinary medicine. Other groups commonly assessed for human medicine but with prospective expansion to animal health are the genera and , which have been demonstrating interesting effects on the prevention of viral diseases, and in dentistry, respectively. Although bacteria from the genera and also have great potential for use in animal production, a complete characterization of the candidate strain must be previously made, due to the existence of pathogenic and/or spoilage variants. It is noteworthy that a growing number of studies have investigated the genus , but still in very early stages. However, the hitherto excellent results endorse its application. In this way, in addition to the fact that bacterial probiotics represent a promising approach to promote productivity increase in animal production, the application of other strains than the traditionally employed genera may allow the exploitation of novel mechanisms and enlighten unexplored possibilities.
PubMed: 38073081
DOI: 10.1017/S1466252323000063 -
Clinical & Experimental Optometry May 2024The pathogenesis of chronic dacryocystitis (CDC) is associated with a variety of bacteria. Investigating microflora has the potential to provide a theoretical basis for...
CLINICAL RELEVANCE
The pathogenesis of chronic dacryocystitis (CDC) is associated with a variety of bacteria. Investigating microflora has the potential to provide a theoretical basis for preventing and treating CDC.
BACKGROUND
16S rRNA sequencing is a sequence-based bacterial analysis. The application of 16S rRNA sequencing in CDC is rarely reported.
METHODS
A case-control study of infected and healthy eyes diagnosed as CDC patients was conducted. Seventy-eight patients were divided into A (conjunctival sac secretions from healthy eyes), B (conjunctival sac secretions from affected eyes), and C (lacrimal sac secretions from affected eyes) groups. The flora of samples was analysed with 16S rRNA sequencing, and the data was analysed using QIIME, R, LefSE and other software. The potential functions were analysed by PICRUSt.
RESULTS
A total of 1440 operational taxonomic units (OTUs) were obtained, 139 specific to group A, 220 specific to group B, and 239 specific to group C. There was no significant difference in α index between the three groups. The beta diversity and grouping analysis data indicated that the three groups of flora were similar in species richness and diversity, but there were some differences in composition. In group A, the abundance of . and . was higher; in group B, the abundance of . was higher; in group C, the abundance of . and . was higher. Six pathways were identified to assess the potential microbial functions.
CONCLUSION
Alterations in the microbiota of the conjunctiva and lacrimal sac are associated with the pathogenesis of CDC, which may provide certain guidance for antibiotic treatment of CDC.
PubMed: 38811366
DOI: 10.1080/08164622.2024.2358246 -
The Journal of Antibiotics Sep 2023The prevalence of antimicrobial-resistant Cutibacterium acnes in acne patients has increased owing to inappropriate antimicrobial use. Commensal skin bacteria may play...
The prevalence of antimicrobial-resistant Cutibacterium acnes in acne patients has increased owing to inappropriate antimicrobial use. Commensal skin bacteria may play an important role in maintaining the balance of the skin microbiome by producing antimicrobial substances. Inhibition of Cu. acnes overgrowth can prevent the development and exacerbation of acne vulgaris. Here, we evaluated skin bacteria with anti-Cu. acnes activity. Growth inhibition activity against Cu. acnes was tested using 122 strains isolated from the skin of healthy volunteers and acne patients. Comparative genomic analysis of the bacterium with or without anti-Cu. acnes activity was conducted. The anti-Cu. acnes activity was confirmed by cloning an identified gene cluster and chemically synthesized peptides. Cu. avidum ATCC25577 and 89.7% of the Cu. avidum clinical isolates (26/29 strains) inhibited Cu. acnes growth. The growth inhibition activity was also found against other Cutibacterium, Lactiplantibacillus, and Corynebacterium species, but not against Staphylococcus species. The genome sequence of Cu. avidum showed a gene cluster encoding a novel bacteriocin named avidumicin. The precursor protein encoded by avdA undergoes post-translational modifications, supposedly becoming a circular bacteriocin. The anti-Cu. acnes activity of avidumicin was confirmed by Lactococcus lactis MG1363 carrying avdA. The C-terminal region of the avidumicin may be essential for anti-Cu. acnes activity. A commensal skin bacterium, Cu. avidum, producing avidumicin has anti-Cu. acnes activity. Therefore, avidumicin is a novel cyclic bacteriocin with a narrow antimicrobial spectrum. These findings suggest that Cu. avidum and avidumicin represent potential alternative agents in antimicrobial therapy for acne vulgaris.
Topics: Humans; Bacteriocins; Propionibacterium acnes; Propionibacteriaceae; Acne Vulgaris
PubMed: 37264118
DOI: 10.1038/s41429-023-00635-w -
Biochimica Et Biophysica Acta.... Jun 2024Acne is a common chronic inflammatory disease of the pilosebaceous unit. Transient receptor potential vanilloid 3 (TRPV3) is an ion channel that is involved in...
Acne is a common chronic inflammatory disease of the pilosebaceous unit. Transient receptor potential vanilloid 3 (TRPV3) is an ion channel that is involved in inflammatory dermatosis development. However, the involvement of TRPV3 in acne-related inflammation remains unclear. Here, we used acne-like mice and human sebocytes to examine the role of TRPV3 in the development of acne. We found that TRPV3 expression increased in the skin lesions of Propionibacterium acnes (P. acnes)-injected acne-like mice and the facial sebaceous glands (SGs) of acne patients. TRPV3 promoted inflammatory cytokines and chemokines secretion in human sebocytes and led to neutrophil infiltration surrounding the SGs in acne lesions, further exacerbating sebaceous inflammation and participating in acne development. Mechanistically, TRPV3 enhanced TLR2 level by promoting transcriptional factor phosphorylated-FOS-like antigen-1 (p-FOSL1) expression and its binding to the TLR2 promoter, leading to TLR2 upregulation and downstream NF-κB signaling activation. Genetic or pharmacological inhibition of TRPV3 both alleviated acne-like skin inflammation in mice via the TLR2-NF-κB axis. Thus, our study revealed the critical role of TRPV3 in sebaceous inflammation and indicated its potential as an acne therapeutic target.
Topics: Toll-Like Receptor 2; Animals; Acne Vulgaris; TRPV Cation Channels; Humans; Mice; Sebaceous Glands; Inflammation; Propionibacterium acnes; Male; NF-kappa B; Signal Transduction; Mice, Inbred C57BL; Female
PubMed: 38648901
DOI: 10.1016/j.bbadis.2024.167195 -
The Spine Journal : Official Journal of... Oct 2023The majority of literature on bacterial flora in the disc stands disadvantaged in utilizing traditional culture methods and targeting a single bacterium, Cutibacterium...
"Are we barking up the wrong tree? Too much emphasis on Cutibacterium acnes and ignoring other pathogens"- a study based on next-generation sequencing of normal and diseased discs.
BACKGROUND
The majority of literature on bacterial flora in the disc stands disadvantaged in utilizing traditional culture methods and targeting a single bacterium, Cutibacterium acnes.
PURPOSE
Our objective was to document the diversity in the bacterial flora between normal and degenerated discs for shortlisting potential pathogens using next-generation genomic tools.
STUDY DESIGN
Experimental case-control study.
METHODS
Researchers employed 16S metagenome sequencing to profile bacterial diversity in magnetic resonance imaging normal healthy discs from brain-dead organ voluntary donors (n=20) and 40 degenerated disc samples harvested during surgery (Modic [MC]=20 and non-Modic [NMC]=20). The V3-V4 region was amplified using universal bacterial primers 341F and 806R, and the libraries were sequenced using Illumina NovoSeq 6000 platform. Statistical significance was set at bacteria with a minimum of 100 operational taxonomic unit (OTU) and present in at least 70% of the samples. The quality check-filtered reads were processed using the QIIME-2 pipeline. The OTU clustering and taxonomic classification were carried out for the merged reads using the Greengenes/SILVA reference database. Validation was done by identification of bacterial metabolites in samples using the liquid chromatography-mass spectrometry approach.
RESULTS
Abundant bacteria differing widely in diversity, as evidenced by Alfa and Beta diversity analysis, were present in all control and degenerative samples. The number of bacterial genera was 27 (14-gram-positive: 13-gram-negative) in the control group, 23 (10-gram-positive: 11-gram-negative) in the Modic group, and 16 (11-gram-positive: 5-gram-negative) in the non-Modic group. In the Modic group, gram-negative bacteria OTUs were found to be predominant (more than 50% of the total bacteria identified), whereas in control and non-Modic groups the OTUs of gram-positive bacteria were predominant. Species-level analysis revealed an abundance of opportunistic gram-negative pathogens like Pseudomonas aeruginosa, Sphingomonos paucibacillus, and Ochrobactrum quorumnocens in the discs with Modic changes, more than in non-Modic discs. The presence of bacterial metabolites and quorum-sensing molecules like N-decanoyl-L-homoserine lactone, 6-hydroxynicotinic acid, 2-aminoacetophenone, 4-hydroxy-3-polyprenylbenzoate, PE (16:1(9Z)/18:0) and phthalic acid validated the colonization and cell-cell communication of bacteria in disc ruling out contamination theory. Cutibacterium acnes was not the predominant bacteria in any of the three groups of discs and in fact was in the 16th position in the order of abundance in the control discs (0.72%), seventh position in the Modic discs (1.41%), and 12th position (0.53%) in the non-Modic discs.
CONCLUSION
This study identified a predominance of gram-negative bacteria in degenerated discs and highlights that Cutibacterium acnes may not be the only degeneration-causing bacteria. This may be attributed to the environment, diet, and lifestyle habits of the sample population. Though the study does not reveal the exact pathogen, it may pave the way for future studies on the subject.
CLINICAL SIGNIFICANCE
These findings invite further investigation into causal relationships of bacterial profile with disc degeneration phenotypes as well as phenotype-driven clinical treatment protocols.
Topics: Humans; Case-Control Studies; Gram-Positive Bacterial Infections; Intervertebral Disc Degeneration; Propionibacterium acnes; High-Throughput Nucleotide Sequencing; Intervertebral Disc
PubMed: 37369253
DOI: 10.1016/j.spinee.2023.06.396 -
Microorganisms Jul 2023Dental caries is a biofilm-mediated, sugar-driven, multifactorial, dynamic disease that results in the phasic demineralization and remineralization of dental hard...
Dental caries is a biofilm-mediated, sugar-driven, multifactorial, dynamic disease that results in the phasic demineralization and remineralization of dental hard tissues. Despite scientific advances in cariology, dental caries remains a severe global concern. The aim of this study was to determine the optimization of microbial and molecular techniques for the detection of cariogenic pathogens in dental caries patients, the prevalence of cariogenic bacteria on the basis of socioeconomic, climatological, and hygienic factors, and in vitro evaluation of the antimicrobial activity of selected synthetic antibiotics and herbal extracts. In this study, oral samples were collected from 900 patients for bacterial strain screening on a biochemical and molecular basis. Plant extracts, such as ginger, garlic, neem, tulsi, amla, and aloe vera, were used to check the antimicrobial activity against the isolated strains. Synthetic antimicrobial agents, such as penicillin, amoxicillin, erythromycin, clindamycin, metronidazole, doxycycline, ceftazidime, levofloxacin, and ciprofloxacin, were also used to access the antimicrobial activity. Among 900 patients, 63% were males and 37% were females, patients aged between 36 and 58 (45.7%) years were prone to disease, and the most common symptom was toothache (61%). For oral diseases, 21% used herbs, 36% used antibiotics, and 48% were self-medicated, owing to sweets consumption (60.66%) and fizzy drinks and fast food (51.56%). (29.11%) and (28.11%) were found as the most abundant strains. Seven bacterial strains were successfully screened and predicted to be closely related to genera , , , , , , and . Among plant extracts, the maximum zone of inhibition was recorded by ginger (22.36 mm) and amla (20.01 mm), while among synthetic antibiotics, ciprofloxacin and levofloxacin were most effective against all microbes. This study concluded that phyto extracts of ginger and amla were considered suitable alternatives to synthetic antibiotics to treat dental diseases.
PubMed: 37630520
DOI: 10.3390/microorganisms11081952 -
Scientific Reports Sep 2023Cutibacterium acnes (C. acnes) is one of the most prevalent bacteria that forms the human skin microbiota. Specific phylotypes of C. acnes have been associated with the...
Cutibacterium acnes (C. acnes) is one of the most prevalent bacteria that forms the human skin microbiota. Specific phylotypes of C. acnes have been associated with the development of acne vulgaris, while other phylotypes have been linked to healthy skin. In this scenario, bacterial extracellular vesicles (EVs) play a role in the interkingdom communication role with the human host. The purpose of this study was to examine the impact of EVs generated by various phylotypes of C. acnes on inflammation and sebum production using different in vitro skin cell types. The main findings of this study reveal that the proteomic profile of the cargo embodied in the EVs reflects distinct characteristics of the different C. acnes phylotypes in terms of life cycle, survival, and virulence. The in vitro skin cell types showed an extended pro-inflammatory modulation of SLST A1 EVs consistently triggering the activation of the inflammation-related factors IL-8, IL-6, TNFα and GM-CSF, in comparison to SLST H1 and SLST H2. Additionally, an acne-prone skin model utilizing PCi-SEB and arachidonic acid as a sebum inducer, was employed to investigate the impact of C. acnes EVs on sebum regulation. Our findings indicated that all three types of EVs significantly inhibited sebum production after a 24-h treatment period, with SLST H1 EVs exhibiting the most pronounced inhibitory effect when compared to the positive control. The results of this study highlight the protective nature of C. acnes SLST H1 EVs and their potential use as a natural treatment option for alleviating symptoms associated with inflammation and oily skin.
Topics: Humans; Proteomics; Skin; Skin Diseases; Extracellular Vesicles; Acne Vulgaris; Propionibacterium acnes; Factor VIII; Inflammation
PubMed: 37749255
DOI: 10.1038/s41598-023-43354-w -
GeroScience Apr 2024Oral health plays a significant role in the quality of life and overall well-being of the aging population. However, age-related changes in oral health are not well...
Oral health plays a significant role in the quality of life and overall well-being of the aging population. However, age-related changes in oral health are not well understood due to challenges with current animal models. In this study, we analyzed the oral health and microbiota of a short-lived non-human primate (i.e., marmoset), as a step towards establishing a surrogate for studying the changes that occur in oral health during human aging. We investigated the oral health of marmosets using cadaveric tissues in three different cohorts: young (aged ≤6 years), middle-aged, and older (>10 years) and assessed the gingival bacterial community using analyses of the V3-V4 variable region of 16S rRNA gene. The oldest cohort had a significantly higher number of dental caries, increased dental attrition/erosion, and deeper periodontal pocket depth scores. Oral microbiome analyses showed that older marmosets had a significantly greater abundance of Escherichia-Shigella and Propionibacterium, and a lower abundance of Agrobacterium/Rhizobium at the genus level. Alpha diversity of the microbiome between the three groups showed no significant differences; however, principal coordinate analysis and non-metric multidimensional scaling analysis revealed that samples from middle-aged and older marmosets were more closely clustered than the youngest cohort. In addition, linear discriminant analysis effect size (LEFSe) identified a higher abundance of Esherichia-Shigella as a potential pathogenic biomarker in older animals. Our findings confirm that changes in the oral microbiome are associated with a decline in oral health in aging marmosets. The current study suggests that the marmoset model recapitulates some of the changes in oral health associated with human aging and may provide opportunities for developing new preventive strategies or interventions which target these disease conditions.
Topics: Humans; Animals; Aged; Middle Aged; Callithrix; Oral Health; RNA, Ribosomal, 16S; Dental Caries; Quality of Life; Aging
PubMed: 37775702
DOI: 10.1007/s11357-023-00939-7