血液因素对龈下菌斑BANA试验结果的影响

目的检测血液因素对慢性牙周炎龈下菌斑BANA试验敏感性、稳定性的影响。方法将不同血液成分及混有健康人血的牙龈卟咻菌菌液分别进行BANA试验。结果各检测稀释滴度的全血、冻融全血、血浆样本BANA试验反应均为阴性;当全血各稀释梯度中牙龈卟啉菌菌量大于10^5时反映结果呈现为BANA试验阳性,胰蛋白酶样酶A600nm值均大于0．20。结论BANA试验的敏感度并不受血液成分混入的影响而降低,证实试验具有较好的稳定性。     

PCR直接检测龈下菌斑主要可疑牙周致病菌

目的：应用PCR方法直接检测龈下菌斑主要可疑牙周致病菌与牙周病活动部位的关系,探讨其方法的可行性并探讨其主要可疑牙周致病菌的分布规律。方法：应用聚合酶链反应(polymerase chain reaction,PCR)直接检测龈下菌斑主要可疑致病菌16s RNA保守区域片段。40名受试者包括牙周病患者20人,每人同口取一个牙周病活动部位,一个相对健康或牙周病静止对照部位;成人健康者20人,每人各取一个标本。结果：龈下菌斑5种可疑牙周致病菌在牙周病活动部位的检出率牙龈卟啉菌为86％,福赛类杆菌为95％,螺旋体为86％,中间普氏菌和黑色普氏菌分别为95％和33％,均显著高于同口部位对照组和健康对照组。结论：PCR直接检测菌斑牙龈卟啉单胞菌、中间普氏菌、福赛类杆菌、齿密螺旋体及黑色普氏菌匀与牙周炎活动部位相关。     

健康青少年口腔正常菌群的初步研究

本文采用非选择性培养基对22名健康青少年的唾液、沟裂菌斑、龈上菌斑及龈下菌斑中的需氧菌、兼性厌氧菌及专性厌氧菌进行了分离培养,并计算其在不同标本中占可培养菌的百分比及检出率。结果共分离到包括18个菌属的35种细菌。其中,链球菌、放线菌、奈瑟氏球菌、二氧化碳噬纤维菌、类杆菌、梭杆菌,奴卡氏菌及棒状杆菌在口腔4个部位的检出率及所占比例均较高,是健康青少年口腔中的优势菌群.通过比较还发现,其中一些菌在口腔4个部位的分布存在一定差异.本文还采用刚果红负性染色涂片法,镜下观察龈上、龈下菌斑中的螺旋体,并计算其相对比例.结果龈下菌斑中螺旋体的相对比例明显高于龈上菌斑.     

牙周炎患者菌斑细菌的药敏分析

目的:了解牙周炎患者龈下菌斑细菌对于口腔常用抗菌药物的敏感性.方法:从慢性牙周炎患者龈下菌斑中分离培养牙周可疑致病菌,测定6种口腔常用抗菌药物对其的最低抑菌浓度,分析以上致病菌对这几类抗菌药物的敏感性.结果:共从50名慢性牙周炎患者牙周袋中分离培养72株革兰阴性厌氧菌进行药物敏感试验.全部实验菌株都对阿莫西林/克拉维酸敏感.对甲硝唑、替硝唑、奥硝唑未见耐药株产生.结论:对于慢性牙周炎患者龈下菌斑中分离的厌氧菌,阿莫西林/克拉维酸和硝基咪唑类药物甲硝唑、替硝唑和奥硝唑的抑菌作用强.     

成人牙周炎龈下套氧菌分离鉴定及药敏试验结果分析

分离并鉴定了329例成人牙周炎龈下优势厌氧菌群,并对不同病程中的菌群变迁、厌氧菌的药物敏感性进行了分析.成人牙周炎龈下标本中厌氧菌阳性检出率为97.9%,其中以牙龈紫质单胞菌检出率最高(38.5%),具核梭杆菌次之(18.9%).随着牙周病变程度的加重,牙龈紫质单胞菌、具核梭杆菌、产黑色素普氏菌、星群厌氧链球菌、厌氧消化链球菌的检出率增高(P＜0.05),小韦荣球菌的检出率下降(P＜0.01),表明前5种厌氧菌在AP发病过程中有重要作用,小韦荣球菌与之无关.替硝唑、甲硝唑和克林霉素对438株革兰氏阴性厌氧菌的MIC90分别为1～8,2～8和4～16 mg/L,对278株革兰氏阳性厌氧菌的MIC90分别为16～32,16～64和4～16 mg/L,表明替硝唑和甲硝唑体外抗革兰氏阴性厌氧菌效果优于克林霉素,抗革兰氏阳性厌氧菌作用不如克林霉素.     

替硝唑在牙周炎治疗中的临床及微生物学观察

本文通过对３０例牙周炎患者在口服替硝唑前后的临床症状及龈下菌斑的暗视野显微镜下螺旋体等菌种比例的变化,观察该药对牙周炎及其致病菌的疗效。结果：服用替硝唑后,在龈下菌斑中螺旋体的比例明显下降,而球菌的比例明显增高。牙龈指数、牙龈出血指数和探针深度均较服药前有明显好转（Ｐ＜０．００１）。提示：替硝唑能有效地抑制和杀灭龈下菌斑中的厌氧菌,并迅速消除牙周炎症,减轻症状。     

牙龈卟啉杆菌W83单克隆抗体的研制与鉴定

牙龈类杆菌曾是重要的产黑色素类杆菌群菌株,最近重新命名为牙龈卟啉杆菌,该菌为牙周病龈下菌斑中关键性厌氧菌,与成人慢性牙周炎的发生、发展有密切关系。作者用牙龈卟啉杆菌侵袭型菌株W83,作为免疫原,通过免疫小鼠、细胞融合、筛选、克隆化,最后得到一株能够稳定分泌抗牙龈卟啉杆菌W83的单克隆抗体杂交瘤细胞系,经鉴定该单抗特异性良好,可用于临床该菌的检出和生态学研究。     

二氧化碳噬纤维菌在牙周炎、牙龈炎患者及健康人口腔中分布的研究

为了解Capno菌与牙周病的关系,本实验分别对25例成人牙周炎、17例牙龈炎患者以及153名健康人(36名儿童,34名青年,49名老年人及34名与牙周病患者对照研究的健康成人)的龈上菌斑、龈下菌斑和唾液标本中的Capno菌的检出结果进行了统计学分析。研究表明:Capno菌在牙周炎、牙龈炎患者及健康人的菌斑及唾液中的检出率无差异(P>0.05),认为该菌是口腔的正常菌群之一。Capno在健康人菌斑中的检出率以儿童、青年为高,可能是青少年口腔中的优势菌群之一。     

口源性口臭患者主要厌氧菌的分布

目的分析口臭患者龈下菌斑和舌苔上主要相关厌氧菌的分布情况。方法选择口腔门诊中口臭患者29例,鼻闻法来确定产臭部位和非产臭部位;分别采集龈下菌斑和舌苔标本接种在非选择性培养基和核梭杆菌选择性培养基,厌氧培养5d后记录非选择性培养基上生长的细菌总数、产黑色素细菌总数及核梭杆菌选择性培养基上生长的目的菌总数。结果29例患者中,15例患者的口臭主要来源于龈缘菌斑,10例主要来源于舌苔,4例患者的口臭由龈缘菌斑和舌苔共同产生;产臭部位和非产臭部位相比,细菌总数、产黑色素菌和具核梭杆菌数都明显上升（P〈0．01）。结论口源性口臭患者口气变化与产黑色素细菌、核梭杆菌相关。     

正常口腔中的可疑牙周致病菌

本实验对49例正常口腔的龈上和龈下菌斑内的可疑牙周致病菌(SPB)的分布情况进行了观察分析。 49例研究对象男21例,女28例;6～25岁,无龋,无牙周病的健康人。三月内未服用抗生素,未接受任何牙周治疗。定对在右上颌第一磨牙(No.3)的龈上及龈下菌斑。用BHI培养基厌氧培养37℃,4天以后进行细菌菌落计数,转种分离和鉴定。龈上菌斑内SPB的检出率按其高低顺序依次为:二氧化碳噬纤维菌、梭杆菌、消化链球菌、唾液弯曲杆菌、产黑色素类杆菌、优杆菌,溶蚀艾肯氏菌、生疾月形单胞菌。二氧化碳噬纤维菌中依次为牙龈二氧化碳噬纤维菌、黄褐二氧化碳噬纤维菌及生疾二氧化碳噬纤     

A Cross-Sectional Survey of Bacterial Species in Plaque from Client Owned Dogs with Healthy Gingiva,Gingivitis or Mild Periodontitis

Periodontal disease is the most widespread oral disease in dogs which if left untreated results in significant pain to the pet and loss of dentition. The objective of this study was to identify bacterial species in canine plaque that are significantly associated with health, gingivitis and mild periodontitis (<25% attachment loss). In this survey subgingival plaque samples were collected from 223 dogs with healthy gingiva, gingivitis and mild periodontitis with 72 to 77 samples per health status. DNA was extracted from the plaque samples and subjected to PCR amplification of the V1-V3 region of the 16S rDNA. Pyrosequencing of the PCR amplicons identified a total of 274 operational taxonomic units after bioinformatic and statistical analysis. Porphyromonas was the most abundant genus in all disease stages, particularly in health along with Moraxella and Bergeyella. Peptostreptococcus, Actinomyces, and Peptostreptococcaceae were the most abundant genera in mild periodontitis. Logistic regression analysis identified species from each of these genera that were significantly associated with health, gingivitis or mild periodontitis. Principal component analysis showed distinct community profiles in health and disease. The species identified show some similarities with health and periodontal disease in humans but also major differences. In contrast to human, healthy canine plaque was found to be dominated by Gram negative bacterial species whereas Gram positive anaerobic species predominate in disease. The scale of this study surpasses previously published research and enhances our understanding of the bacterial species present in canine subgingival plaque and their associations with health and early periodontal disease.     

The subgingival microbiome of clinically healthy current and never smokers

Dysbiotic oral bacterial communities have a critical role in the etiology and progression of periodontal diseases. The goal of this study was to investigate the extent to which smoking increases risk for disease by influencing the composition of the subgingival microbiome in states of clinical health. Subgingival plaque samples were collected from 200 systemically and periodontally healthy smokers and nonsmokers. 16S pyrotag sequencing was preformed generating 1 623 713 classifiable sequences, which were compared with a curated version of the Greengenes database using the quantitative insights into microbial ecology pipeline. The subgingival microbial profiles of smokers and never-smokers were different at all taxonomic levels, and principal coordinate analysis revealed distinct clustering of the microbial communities based on smoking status. Smokers demonstrated a highly diverse, pathogen-rich, commensal-poor, anaerobic microbiome that is more closely aligned with a disease-associated community in clinically healthy individuals, suggesting that it creates an at-risk-for-harm environment that is primed for a future ecological catastrophe.     

Oral Disease in Animals: The Australian Perspective. Isolation and Characterisation of Black-Pigmented Bacteria from the Oral Cavity of Marsupials

A wide range of animals suffer from periodontal disease. However, there is very little reported on disease and oral micro-biota of Australian animals. Therefore, the oral cavity of 90 marsupials was examined for oral health status. Plaque samples were collected from the subgingival margins using curettes or swabs. Plaque samples were plated onto non-selective trypticase soy agar plates, selective trypticase soy agar, non-selective and selective Wilkens Chalgrens Agar. Plates were incubated in an anaerobic atmosphere and examined after 7–14 days for the presence of black–brown-pigmented colonies. A combination of morphological and biochemical tests were used (colonial morphology, pigmentation, aerobic growth, Gram reaction, fluorescence under long-wave UV light (360 nm), production of catalase, enzymatic activity with fluorogenic substrates and haemagglutination of sheep red cells) to identify these organisms. Black-pigmented bacteria were cultivated from the plaque of 32 animals including six eastern grey kangaroos, a musky rat kangaroo, a whiptail and a red-necked wallaby, 18 koalas, a bandicoot and five brushtail possums. No black-pigmented colonies were cultivated from squirrel or sugar gliders or quokkas or from marsupial mice. The majority of isolates were identified as Porphyromonas gingivalis -like species with the higher prevalence of isolation from the oral cavity of macropods (the kangaroos and wallabies). Oral diseases, such as gingivitis can be found in native Australian animals with older koalas having an increase in disease indicators and black-pigmented bacteria. Non-selective Wilkens Chalgren Agar was the medium of choice for the isolation of black-pigmented bacteria.     

Effect of the Conditions of Incubation in the Recovery of Oral Actinomyces

The Gram-positive Pleomorphic Bacilli (GPPB), especially the genus Actinomyces, are part of the oral microflora and they are generally associated with cement caries. They are also found in inactive sites in periodontal disease and in pulpar infections. The aim of the present work was to find an appropriate isolation culture medium for the recovery of the genus Actinomyces from oral samples, and to propose a minimum schema of biochemical tests for the identification and differentiation of Actinomyces species from other GPPB. Samples of saliva, subgingival plaque and post-extraction acute alveolar osteitis were cultivated in Starch Casein Agar (SCA), Yeast Extract Dextrose Agar (YDA), Columbia Blood Agar 5% (BA), both with cephadroxyl (10 μL/mL) and without antibiotic. The plates were incubated in aerobic conditions, in strict anaerobic conditions and in a candle jar for 5 days at 37°C. (1) The highest recovery of Actinomyces was obtained in BA without antibiotic in a candle jar for all oral samples. (2) In all the incubated in SCA, YDA and BA with antibiotic,Actinomyces , other GPPB, Gram-positive and -negative cocci and Gram-negative bacilli showed growth in all aerobic conditions. (3) In BA medium with antibiotic the Gram-negative microflora was inhibited, the Actinomyces recovery being lower than in the other media studied.     

应用16S rDNA检测固定矫治患者牙周可疑病原菌变化研究

应用16S rDNA检测固定矫治患者龈下菌斑中牙周可疑病原,探讨戴用固定矫治器对牙周组织健康的影响。随机选择36例治疗时间超过6个月的固定矫治患者组成实验组,29例未经正畸治疗者组成对照组。分别检验特定部位牙周临床指数并收集龈下菌斑样本。采用16S rDNA检测9种牙周可疑致病菌。实验组与对照组相比牙龈指数、牙周袋深度、探诊出血差异有明显统计学意义(P<0.05);牙周可疑病原菌中牙龈卟啉菌、齿垢密螺旋体在实验组的检出率明显高于对照组(P<0.05)。固定矫治器戴入可引起患者牙周可疑病原菌的明显变化。     

Periodontal pathogens: a quantitative comparison of anaerobic culture and real-time PCR

Periodontitis is a multi-factorial chronic inflammatory and destructive disease of the tooth-supporting tissues. Quantitative anaerobic culture techniques have been used for microbial diagnosis of the different forms of the disease. The aim of this study was to compare real-time PCR with quantitative anaerobic culture for detection and quantification of 5 prominent periodontal pathogens. Real-time PCR assays with the 16s rRNA genes of Actinobacillus actinomycetemcomitans, Prevotella intermedia, Tannerella forsythensis, Peptostreptococcus micros and Fusobacterium spp. were developed. The PCR was validated on pure cultures of various bacterial strains. Subsequently, subgingival plaque samples from 259 adult patients with periodontitis were analyzed with quantitative anaerobic culture and real-time PCR. A standard curve for DNA quantification was created for each primer-probe set based on colony-forming units equivalents. All bacterial species were correctly identified. The lower limits of detection by PCR varied between 1-50 colony-forming units equivalents depending on the species. No cross-reactivities with heterologous DNA of other bacterial species were observed. Real-time PCR results showed a high degree of agreement with anaerobic culture results. Real-time PCR is a reliable alternative for diagnostic quantitative anaerobic culture of subgingival plaque samples.     

Pigmented Prevotella species in the periodontally healthy oral cavity

Abstract Pigmented Prevotella spp. have been connected with oral infections as well as being part of the healthy gingival flora. The aim of this study was to determine the presence of pigmented Prevotella spp. in saliva and gingival crevice samples from periodontally healthy adults. Twelve Caucasian female subjects (mean age 28 years, range 21–36 years) with no pockets ≥ 4 mm, nor bleeding after probing were selected for this study. Paraffin-stimulate saliva was collected first; then, a pooled subgingival bacterial sample was taken with a sterile curette from mesiobuccal surfaces of all first molars. The samples were inoculated on to non-selective and selective media and incubated anaerobically. The most frequent species isolated were Pr. melaninogenica, Pr. intermedia and Pr. loescheii , found in 11, ten and nine subjects, respectively. The mean percentages of the total cultivable anaerobic microflora in salivary/subgingival samples were 14.7/0.6 for Pr. melaninogenica , 3.1/5.3 for Pr. intermedia and 2.6/1.2 for Pr. loescheii. Pr. denticola was found in one saliva sample and Pr. corporis , in two subgingival samples only. The number of different pigmented Prevotella spp. in the same mouth was 2–4 (mean 2.75). In conclusion, Pr. melaninogenica, Pr. intermedia and Pr. loescheii seem to be common microorganisms in the periodontally healthy oral cavity.     

Oral microbiota of children in a school-based dental clinic

ObjectivesDental caries disproportionately affects disadvantaged subjects. This study hypothesized that there were greater caries extent and higher levels of caries-associated and anaerobic subgingival bacterial species in oral samples of Hispanic and immigrant children compared with non-Hispanic and US born children.MethodsChildren from a school-based dental clinic serving a community with a large Hispanic component were examined, and the extent of caries was recorded. Microbial samples were taken from teeth and the tongues of children. Samples were analyzed using DNA probes to 18 oral bacterial species.ResultsSeventy five children were examined. Extent of caries increased with child age in immigrant, but not in US born or Hispanic children. There were no differences in the microbiota based on ethnicity or whether the child was born in US or not. There was a higher species detection frequency from teeth than tongue samples. Levels of Streptococcus mutans and other Streptococcus spp increased with caries extent. Prevotella intermedia, Tannerella forsythia and Selenomonas spp were detected at low levels in these children.ConclusionsWe conclude that, while there was a high rate of dental caries in disadvantaged school children, there were no differences in the caries-associated microbiota, including S. mutans, based on ethnicity or immigration status. Furthermore, while anaerobic subgingival, periodontal pathogens were also detected in children, there was no difference in species detection based on ethnicity or immigration status. Increased levels of streptococci, including S. mutans, however, were detected with high caries levels. This suggested that while it is beneficial to target preventive and treatment programs to disadvantaged populations, there is likely no additional benefit to focus on subgroups within a population already at high risk for dental disease.