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Veterinary and Animal Science Dec 2019Periodontal disease is the most common infectious disease of cats and dogs which are strongly associated with periodontal pathogens. The primary etiologic factor in the...
Periodontal disease is the most common infectious disease of cats and dogs which are strongly associated with periodontal pathogens. The primary etiologic factor in the formation of periodontal disease is microbial dental plaque accumulation on teeth. In our research, we aimed to investigate the presence of periodontal disease-related bacterial species in dental plaques of cats and dogs. Specimens collected from 50 cats and 51 dogs with periodontal disease examined in terms of periodontal pathogens by polymerase chain reaction (PCR) using primers directed to 16S rRNA and tdpA genes. Our findings indicate the presence of periodontal disease-related pathogens, especially (cats 96%, dogs 88%), (cats 90%, dogs 57%) and, (cats 70%, dogs 39%). In addition, the prevalence of (cats 2%, dogs 4%) well-known pathogen in cats and dogs were isolated with an extremely low percentage. Furthermore, our results suggest that the feline oral cavity microbiota has considerably more diversity than dogs. Consequently, daily oral hygiene practices may become essential for controlling the pathogenic bacteria which have clinical importance and in preventing the propagation of microorganisms in the oral cavity of cats and dogs.
PubMed: 32734086
DOI: 10.1016/j.vas.2019.100069 -
Mikrobiyoloji Bulteni Apr 2020Prevotella species, being members of the human microbiota, are obligate anaerobic gram-negative bacteria. These organisms may cause opportunistic infections, including...
Prevotella species, being members of the human microbiota, are obligate anaerobic gram-negative bacteria. These organisms may cause opportunistic infections, including specific oral infections, local or systemic infections. A significant increase of resistance to some antimicrobials has been detected among Prevotella species. The frequency of resistance vary among isolates from different infection sources and between geographic locations. The knowledge about the antimicrobial susceptibility patterns of different Prevotella species is limited in Turkey. Providing the antimicrobial susceptibility data of these bacteria is very important for effective empirical treatment. In this study, we aimed to determine susceptibility data for 12 antimicrobial agents against Prevotella strains originating from human infections, collected in two centers in Turkey. A total of 118 Prevotella strains, isolated from different clinical samples in Marmara University Faculty of Medicine Medical Microbiology and Istanbul University Faculty of Dentistry Oral Microbiology Laboratories between January 2014-December 2017, were tested. Organisms were identified by using MALDI-TOF MS and by 16S rRNA gene sequencing. Minimal inhibitor concentrations of ampicillin, ampicillin-sulbactam, piperacillin-tazobactam, cefoxitin, meropenem, imipenem, clindamycin, tetracycline, tigecycline, moxifloxacin and metronidazole were determined using gradiyent test methodology (E-test; bioMerieux, France) and the European Committee on Antimicrobial Susceptibility Testing, Clinical and Laboratory Standards Institute and Food and Drug Administration guidelines were used for interpretation. Thirteen different Prevotella species were identified, Prevotella bivia and Prevotella nigrescens were the most prevalent species (n= 21) followed by Prevotella buccae (n= 19). All Prevotella strains were susceptible to piperacillin-tazobactam, cefoxitin, meropenem, imipenem and tigecycline. A total of 2 (1.7%) isolates were resistant to metronidazole and 1 (0.8%) isolate was intermediately resistant to ampicillin/sulbactam. The frequency of resistant isolates against ampicillin, clindamycin, tetracycline and moxifloxacin were 57.6%, 36.4%, 18% and 16.3%, respectively. In conclusion, piperacillin/tazobactam, cefoxitin, and tigecycline displayed high in vitro activity against Prevotella spp. and they all remained good candidates for empiric therapy. Imipenem and meropenem were also found to be very active, but the usage of carbapenems should be reserved for serious mixed infections, potentially accompanied by other resistant organisms. Intermediate resistance to ampicillinsulbactam and the resistance against metronidazole emphasized the need of periodic monitoring of their susceptibility patterns. The high rates of non-susceptibility to ampicillin, clindamycin, tetracycline and moxifloxacin indicated that these antimicrobials should not be used for treatment of infections without prior antimicrobial susceptibility testing.
Topics: Anti-Bacterial Agents; Bacteria, Anaerobic; Bacteroidaceae Infections; Humans; Microbial Sensitivity Tests; Prevotella; RNA, Ribosomal, 16S; Turkey
PubMed: 32723280
DOI: 10.5578/mb.69309 -
PloS One 2020The study of oral disease progression, in relation to the accumulation of subgingival biofilm in gingivitis and periodontitis is limited, due to either the ability to...
The study of oral disease progression, in relation to the accumulation of subgingival biofilm in gingivitis and periodontitis is limited, due to either the ability to monitor plaque in vitro. When compared, optical spectroscopic techniques offer advantages over traditional destructive or biofilm staining approaches, making it a suitable alternative for the analysis and continued development of three-dimensional structures. In this work, we have developed a confocal Raman spectroscopy analysis approach towards in vitro subgingival plaque models. The main objective of this study was to develop a method for differentiating multiple oral subgingival bacterial species in planktonic and biofilm conditions, using confocal Raman microscopy. Five common subgingival bacteria (Fusobacterium nucleatum, Streptococcus mutans, Veillonella dispar, Actinomyces naeslundii and Prevotella nigrescens) were used and differentiated using a 2-way orthogonal Partial Least Square with Discriminant Analysis (O2PLS-DA) for the collected spectral data. In addition to planktonic growth, mono-species biofilms cultured using the 'Zürich Model' were also analyzed. The developed method was successfully used to predict planktonic and mono-species biofilm species in a cross validation setup. The results show differences in the presence and absence of chemical bands within the Raman spectra. The O2PLS-DA model was able to successfully predict 100% of all tested planktonic samples and 90% of all mono-species biofilm samples. Using this approach we have shown that Confocal Raman microscopy can analyse and predict the identity of planktonic and mono-species biofilm species, thus enabling its potential as a technique to map oral multi-species biofilm models.
Topics: Actinomyces; Bacteria; Bacteriological Techniques; Biofilms; Culture Media; Fusobacterium nucleatum; Gingiva; Gingivitis; Microbial Viability; Microbiota; Microscopy, Confocal; Nonlinear Optical Microscopy; Periodontitis; Plankton; Prevotella intermedia; Streptococcus mutans; Veillonella
PubMed: 32392236
DOI: 10.1371/journal.pone.0232912 -
Microorganisms May 2020Oral bacteria possess the ability to form biofilms on solid surfaces. After the penetration of oral bacteria into the pulp, the contact between biofilms and pulp tissue...
Oral bacteria possess the ability to form biofilms on solid surfaces. After the penetration of oral bacteria into the pulp, the contact between biofilms and pulp tissue may result in pulpitis, pulp necrosis and/or periapical lesion. Depending on the environmental conditions and the availability of nutrients in the pulp chamber and root canals, mainly Gram-negative anaerobic microorganisms predominate and form the intracanal endodontic biofilm. The objective of the present study was to investigate the role of different substrates on biofilm formation as well as the separate and collective incorporation of six endodontic pathogens, namely and into a nine-species "basic biofilm". This biofilm was formed in vitro as a standard subgingival biofilm, comprising and The resulting endodontic-like biofilms were grown 64 h under the same conditions on hydroxyapatite and dentin discs. After harvesting the endodontic-like biofilms, the bacterial growth was determined using quantitative real-time PCR, were labeled using fluorescence in situ hybridization (FISH) and analyzed by confocal laser scanning microscopy (CLSM). The addition of six endodontic pathogens to the "basic biofilm" induced a decrease in the cell number of the "basic" species. Interestingly, counts increased in biofilms containing and respectively, both on hydroxyapatite and on dentin discs, whereas counts increased only on dentin discs by addition of . The growth of on hydroxyapatite discs and of and on dentin discs were significantly higher in the biofilm containing all species than in the "basic biofilm". Contrarily, the counts of , and on hydroxyapatite discs as well as counts of and on dentin discs decreased in the all-species biofilm. Overall, all bacterial species associated with endodontic infections were successfully incorporated into the standard multispecies biofilm model both on hydroxyapatite and dentin discs. Thus, future investigations on endodontic infections can rely on this newly established endodontic-like multispecies biofilm model.
PubMed: 32384777
DOI: 10.3390/microorganisms8050674 -
Journal of Alzheimer's Disease : JAD 2020Microbial agents including periodontal pathogens have recently appeared as important actors in Alzheimer's disease (AD) pathology. We examined associations of clinical...
Microbial agents including periodontal pathogens have recently appeared as important actors in Alzheimer's disease (AD) pathology. We examined associations of clinical periodontal and bacterial parameters with incident all-cause and AD dementia as well as AD mortality among US middle-aged and older adults. Clinical [Attachment Loss (AL); probing pocket depth (PPD)] and bacterial [pathogen immunoglobulin G (IgG)] periodontal markers were investigated in relation to AD and all-cause dementia incidence and to AD mortality, using data from the third National Health and Nutrition Examination Surveys (NHANES III, 1988-1994) linked longitudinally with National Death Index and Medicare data through January 1, 2014, with up to 26 years of follow-up. Sex- and age-specific multivariable-adjusted Cox proportional hazards models were conducted. Among those ≥65 years, AD incidence and mortality were consistently associated with PPD, two factors and one cluster comprised of IgG titers against Porphyromonas gingivalis (P. gingivalis), Prevotella melaninogenica (P. melaninogenica) and Campylobacter rectus (C. rectus) among others. Specifically, AD incidence was linked to a composite of C. rectus and P. gingivalis titers (per SD, aHR = 1.22; 95% CI, 1.04-1.43, p = 0.012), while AD mortality risk was increased with another composite (per SD, aHR = 1.46; 95% CI, 1.09-1.96, p = 0.017) loading highly on IgG for P. gingivalis, Prevotella intermedia, Prevotella nigrescens, Fusobacterium nucleatum, C. rectus, Streptococcus intermedius, Capnocylophaga Ochracea, and P. melaninogenica. This study provides evidence for an association between periodontal pathogens and AD, which was stronger for older adults. Effectiveness of periodontal pathogen treatment on reducing sequelae of neurodegeneration should be tested in randomized controlled trials.
Topics: Aged; Alzheimer Disease; Campylobacter rectus; Dementia; Female; Health Surveys; Humans; Incidence; Male; Middle Aged; Periodontitis; Porphyromonas gingivalis; Prevotella melaninogenica; United States
PubMed: 32280099
DOI: 10.3233/JAD-200064 -
Journal of Indian Society of... 2020Bacterial colonization of dentition in different age groups can impact prognosis in different dental diseases. Latest diagnostic technique such as matrix-assisted laser...
Early and accurate detection of bacterial isolates from dental plaque in subjects with primary, mixed, and permanent dentition by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry technique.
BACKGROUND
Bacterial colonization of dentition in different age groups can impact prognosis in different dental diseases. Latest diagnostic technique such as matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF) is increasingly being used for accurate identification of bacteria. This study was undertaken to evaluate the MALDI-TOF MS technique to identify bacterial pathogens from dental plaques in subjects with primary, mixed, and permanent dentition.
MATERIALS AND METHODS
The study included 150 subjects of different age groups and were divided into three groups - Group A: Subjects with primary dentition ( = 50), Group B: Subjects with mixed dentition ( = 50), and Group C: Subjects with permanent dentition ( = 50). Subgingival dental plaque samples were collected from buccal and lingual surfaces of premolar and molar teeth. Clinical parameters such as gingival index were recorded. Samples were cultured in routine aerobic and anaerobic medium. Bacterial growths were assessed by semiquantitative methods. Bacterial isolates were confirmed by MALDI-TOF MS technique.
RESULTS
MALDI-TOF MS detected all the culture-grown bacteria. In primary dentition group, purple and yellow complex bacteria predominated. spp. was the predominant bacteria (51%) followed by (19%) and spp. (19%). In mixed dentition and permanent group also, spp. was predominant (46%) followed by spp. (24%) and (19%). However, in both groups, orange complex bacteria (bridge complex) such as and red complex bacteria (, 3%) were seen. For majority of bacteria, the load increased with age.
CONCLUSIONS
The bacterial isolates showed a distinct age-specific colonization. The use of advanced technique such as MALDI-TOF MS is helpful in the detection of periodontal pathogens, and the effective oral health programs can be implemented to minimize the risk of periodontal diseases.
PubMed: 32189836
DOI: 10.4103/jisp.jisp_303_19 -
Antibiotics (Basel, Switzerland) Feb 2020The in vitro resistance of selected red/orange complex periodontal pathogens to tinidazole was compared with four other antibiotics. Subgingival biofilm samples from 88...
The in vitro resistance of selected red/orange complex periodontal pathogens to tinidazole was compared with four other antibiotics. Subgingival biofilm samples from 88 adults with severe periodontitis were anaerobically incubated on enriched Brucella blood agar with and without supplementation with tinidazole (16 mg/L), metronidazole (16 mg/L), amoxicillin (8 mg/L), doxycycline (4 mg/L), or clindamycin (4 mg/L). Growth of , , or on antibiotic-supplemented plates indicated their in vitro antibiotic resistance. Tinidazole inhibited all test species, except , , and in 3.8%, 10.2%, and 88.9% of species-positive patients, respectively. Significantly fewer patients yielded tinidazole-resistant test species, and had significantly lower subgingival proportions of tinidazole-resistant organisms, than patients with amoxicillin, doxycycline, or clindamycin-resistant species, but not those with metronidazole-resistant strains. Joint in vitro species resistance to tinidazole and amoxicillin, or metronidazole and amoxicillin, was rare. Tinidazole performed in vitro similar to metronidazole, and markedly better than amoxicillin, doxycycline, or clindamycin, against fresh clinical isolates of red/orange complex periodontal pathogens. As a result of its similar antimicrobial spectrum, and more convenient once-a-day oral dosing, tinidazole should be considered in place of metronidazole for systemic periodontitis drug therapy.
PubMed: 32046045
DOI: 10.3390/antibiotics9020068 -
Annals of Dermatology Feb 2020Associations between acne and gastrointestinal comorbidities suggest that microbial dysbiosis and intestinal permeability may promote inflammatory acne, a condition...
BACKGROUND
Associations between acne and gastrointestinal comorbidities suggest that microbial dysbiosis and intestinal permeability may promote inflammatory acne, a condition often managed with oral antibiotics.
OBJECTIVE
We performed a case-control study to investigate the skin and gut microbiota in 8 acne patients before and after receiving oral minocycline compared to controls matched by age ±5 years, sex, and race.
METHODS
DNA was extracted from stool samples and facial skin swabs. Sequencing of the V3V4 region of the bacterial 16S rRNA gene was performed using Illumina MiSeq and analyzed using QIIME/MetaStats 2.0 software.
RESULTS
Acne patients included 7 female and 1 male, ages 20~32. Shannon diversity was not significantly different between the skin (=0.153) or gut (<0.999) microbiota of acne patients before and after antibiotics. The gut microbiota in pre-antibiotic acne patients compared to acne-free controls was depleted in probiotics (=0.001), (=0.001), and (=0.026). After antibiotics, the gut microbiota of acne patients was depleted in (=0.001), (=0.002), (=0.010), and (=0.042), while the skin microbiota was enriched in probiotics (=0.028) and (=0.029) and depleted in (=0.009) and (=0.028). At the phylum level, significant enrichment of Bacteroidetes in stool of acne patients following antibiotic treatment (=0.033) led to a decreased Firmicutes to Bacteroidetes ratio.
CONCLUSION
Minocycline produces significant derangements in the microbiota of the skin and gut, including many probiotic species, highlighting the potential for more targeted antimicrobial treatments for acne.
PubMed: 33911705
DOI: 10.5021/ad.2020.32.1.21 -
Journal of Cystic Fibrosis : Official... Mar 2020Prevotella spp. are frequently identified in Cystic Fibrosis sputum. This study examined whether infection with Prevotella nigrescens, a frequently identified member of...
Prevotella spp. are frequently identified in Cystic Fibrosis sputum. This study examined whether infection with Prevotella nigrescens, a frequently identified member of this species, contributes to inflammation in CF bronchial epithelial cells through activation of TLR- and NF-κB signalling pathways. CFBE41o- cells were infected with either P.nigrescens or Pseudomonas aeruginosa and incubated under anaerobic conditions for 4h. P.nigrescens activated TLR2 signalling but not TLR4 signalling while P.aeruginosa activated TLR4 signalling with a lesser effect on TLR2. P.aeruginosa induced significant IκBα phosphorylation 10min post infection with a return to control levels by 30min post infection. A significant induction in nuclear p65 DNA binding was observed at 2h post infection. In contrast, infection with P.nigrescens induced phosphorylation of IκBα 120min post infection, with significant induction in nuclear p65 DNA binding at 4h post infection only. Cytokine gene and protein responses were lower for P.nigrescens compared to P.aeruginosa. This study demonstrates the ability of a clinical P.nigrescens isolate to provoke a delayed NF-κB(p65) driven response through induction in TLR2 signalling and activation of sustained levels of IKKα.
Topics: Bacteria, Anaerobic; Cells, Cultured; Cystic Fibrosis; Host-Pathogen Interactions; Humans; Inflammation; NF-kappa B; Prevotella nigrescens; Pseudomonas aeruginosa; Respiratory Mucosa; Signal Transduction; Toll-Like Receptor 2; Toll-Like Receptor 4; Transcription Factor RelA
PubMed: 31607634
DOI: 10.1016/j.jcf.2019.09.005 -
International Journal of Environmental... Aug 2019Type-2-Diabetes (T2D) and Periodontitis are major inflammatory diseases. However, not much is known about the specific subgingival microbiota in Mexicans with diabetes...
BACKGROUND
Type-2-Diabetes (T2D) and Periodontitis are major inflammatory diseases. However, not much is known about the specific subgingival microbiota in Mexicans with diabetes and metabolic dysbiosis. The aim of this study was to describe the subgingival microbiota of Mexicans with T2D and the different periodontal and metabolic conditions, through "Checkerboard" DNA-DNA hybridization.
METHODS
Subjects were divided into two groups-periodontal-health (PH) (PH_non-T2D; = 59, PH_T2D; = 14) and generalized-periodontitis (GP) (GP_non-T2D; = 67, GP_T2D; = 38). Obesity (BMI ≥ 30 kg/m) and serum levels of glycated-hemoglobin (HbA1c), total-lipids, triglycerides, total-cholesterol, high-density-lipids, and low-density-lipids were measured for the T2D individuals. Subgingival microbial identification was processed for 40 species through DNA-probes.
RESULTS
Subjects with T2D harbored significantly higher mean total levels (PH: < 0.001, and GP_NS), a lower proportion of "red" complex (GP: < 0.01), a higher proportion of "yellow" (GP; < 0.001), and "orange" (GP; < 0.01) complex than the non-T2D. GP_T2D individuals exhibited a greater proportion of putative-species- and ( < 0.001), and and ( < 0.01), than GP_non-T2D. T2D individuals with HbA1c > 8% had presented significantly higher mean pocket-depth and higher levels of ( < 0.05) and those with obesity or dyslipidemia harbored higher levels, prevalence, or proportion of sp., sp., and sp.
CONCLUSIONS
T2D individuals harbored a particular microbial profile different to non-T2D microbiota. Metabolic control was related to dysbiosis of microbiota-HbA1c>8% related to periodontitis and obesity or dyslipidemia with the predominance of saccharolytic bacteria, irrespective of their periodontal condition.
Topics: Adult; Bacteria; Campylobacter; Diabetes Mellitus, Type 2; Female; Gingival Diseases; Humans; Male; Mexico; Microbiota; Middle Aged; Nucleic Acid Hybridization; Periodontitis
PubMed: 31480468
DOI: 10.3390/ijerph16173184