健康青少年口腔优势菌对牙龈卟啉单胞菌拮抗作用的实验研究

目的分析健康青少年口腔优势菌（简称S．t2003）与牙龈卟啉单胞菌的相互关系。方法运用定量混合培养技术,以血链球菌标准株和变形链球菌作为对照。结果S．t2003与血链球菌标准株都对牙龈卟啉单胞菌有很强的抑制作用,但S．t2003的抑菌强度强于血链球菌的标准株;另外S．t2003和血链球菌标准株对牙龈卟啉单胞菌的抑制作用随其相对浓度的变化而改变,显示了S．t2003和血链球菌标准株与牙龈卟啉单胞菌的相互拮抗的关系。结论S．t2003对牙龈卟啉单胞菌具有较强的拮抗作用。     

血链球菌在不同牙周状态下的分布及相关研究

目的：探讨口腔主要过氧化氢产生菌血链球菌和口腔链球菌在不同牙周健康状态下龈下菌群中的分布,及与牙周健康状态和牙龈卟啉单胞菌群中分布的相互关系。方法：纳入符合标准的受试者30人,受试位点86个,其中健康组11人,位点30个,龈炎组9人,位点29个,慢性牙周炎组10人,位点27个,检查记录牙周健康状态[包括牙龈指数（GI）和牙周袋深度（PD）],采集龈下菌斑标本,经厌氧菌培养基和AP-PCR及PCR鉴定后,将各受试组进行比较分析。结果：共获得草绿色链球菌523株,产黑色素菌241株。经AP-PCR及PCR鉴定后,得到血链球菌112株,口腔链球菌56株,牙龈卟啉单胞菌84株,健康组龈下菌斑中血链球菌,口腔链球菌和牙龈卟啉单胞菌构成比与牙周炎组相比有差异显著性;血链球菌和口腔链球菌与GI、PD呈负相关,牙龈卟啉菌与GI、PD呈正相关;血链球菌的构成与牙龈卟啉单胞菌的构成比呈负相关。结论：血链球菌等过氧化氢产生菌在龈下菌斑中比例的下降。可能是微生态失衡,致病菌过度增殖的重要机制。     

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.     

健康青少年口腔优势菌产生过氧化氢的初步研究

目的研究健康青少年口腔优势菌产生过氧化氢的能力。方法采用过氧化物酶法,在体外测得健康青少年口腔优势菌产生过氧化氢的量。结果在需氧条件下,能产生过氧化氢,产生的量与细菌的生长速度成正比;在厌氧条件下,未测得过氧化氢的产生。结论健康青少年口腔优势菌产生过氧化氢的能力强于S.sATCC10556(P0.05)     

老年上呼吸道微生态研究

目的通过研究健康老年机体上呼吸道微生态菌群构成,了解老龄人群上呼吸道优势菌群种类及数量,为从微生态学角度预防呼吸道感染的发生提供参考。方法自沈阳市选取年龄为65～70岁健康老人为研究对象,无菌咽拭法采集咽后壁粘膜表面标本,对菌群进行定量分析、定性鉴定。结果老年人群咽后壁需氧菌群与厌氧菌群数量之比为2.65∶1,检出率较高的需氧菌及厌氧菌包括唾液链球菌、口腔链球菌、缓症链球菌、微黄奈瑟菌、灰色奈瑟菌、麻疹孪生球菌等,检出率较高的菌构成比也较高。结论健康老年人群中检出率和构成比都较高的需氧菌和厌氧菌共同构成老年人上呼吸道口咽部优势菌群,在维持呼吸系统微生态平衡中起重要作用,可作为上呼吸道微生态学研究的重要指标菌。     

不同牙周状态下分离的血链球菌和口腔链球菌H2O2产生能力的比较

目的：比较研究不同牙周状态下分离的血链球菌和口腔链球菌过氧化氢（H2O2）生产能力。方法：随机选出不同牙周状态下分离的血链球菌和口腔链球菌各30株,采用酚红还原-微量板法测定厌氧条件下两种细菌H2O2的产量。结果：血链球菌各组和口腔链球菌各组H2O2的产量差异未见显著性;口腔链球菌产生H2O2的能力强于血链球菌。结论：提示牙周病变部位与健康部位的血链球菌和口腔链球菌产生H2O2的能力基本相似,是否存在基因型的差异尚待进一步研究。     

口腔血链球菌质粒的初步研究

口腔血链球菌是牙周主要有益菌,质粒是细胞染色体外辅助性遗传物质可以赋予细胞不同表型。本研究目的提检测血链球菌质粒ＤＮＡ存在情况,鉴定其分子大小,初步了解血链菌质粒与耐药性的关系,为今后更深入地在分子水平研究血链菌奠定实验基础。     

血链球菌与疾病研究进展

血链球菌是早期定植在口腔内的细菌之一,也是口腔内的常驻菌.血链球菌作为牙周有益菌对大多数牙周可疑致病菌具有拮抗作用.其主要机制为产生过氧化氢和血链素.由于各种原因进入血液循环后可引起感染性心内膜炎,主要机制为血链球菌的表面抗原使血小板产生黏附、聚集,其在血小板表面的结合位点在血小板膜糖蛋白Ib附近.单核细胞在血链球菌的刺激下表达大量的组织因子,激活外源性凝血途径形成血栓.     

低龄儿童龋病牙菌斑的菌群结构分析

目的通过低龄龋齿儿童与口腔健康儿童的微生物组检测,探讨低龄儿童龋病牙菌斑的菌群组成特征。方法在深圳地区招募轻度龋齿、重度龋齿及口腔健康儿童共100名,采集其牙菌斑或健康牙齿表面微生物样本,采用MiSeq测序平台进行16S V4-V5区域的高通量测序。同时,通过生物信息分析方法,对低龄龋齿儿童的牙菌斑的微生物组成的特征进行挖掘分析。结果相对口腔健康儿童,重度龋齿患儿的牙菌斑微生物多样性增加,轻度龋齿患儿则减少,但变化不显著。门水平的优势菌主要为放线菌门、变形菌门及拟杆菌门,虽有相对丰度变化但差异无统计学意义。前10位的优势菌属中,轻度及重度龋齿儿童的最高丰度优势菌属为棒状杆菌属,重度龋齿儿童与健康儿童中的棒状杆菌丰度差异有统计学意义(t=-2.195 5, P=0.028 1)。健康儿童口腔的优势菌属包含了卟啉单胞菌属,但在轻度及重度龋齿儿童的优势菌属则替换为月形单胞菌属。种水平的菌群结构分析显示,前10位的优势菌丰度变化差异无统计学意义,而低丰度的致病菌如变异链球菌(H=27.302 1, P<0.000 1)、Atopobium parvulum(H=17.418 2, P=0.000 2)、栖牙普氏菌(H=10.598 9, P=0.005 0)、唾液普氏菌(H=10.035 0, P=0.006 6)等则在重度龋齿儿童中显著富集。结论低龄龋齿儿童的牙菌斑微生物结构发生了改变,部分口腔有害菌的丰度显著增加与龋病严重程度密切相关。     

伊犁黑蜂蜂胶对口腔主要致龋细菌及生物膜生长的实验研究

目的研究伊犁黑蜂蜂胶对口腔常见致龋细菌及其生物膜生长的影响,观察其防龋效果。方法 (1)通过液体稀释法测定伊犁黑蜂蜂胶对口腔常见致龋菌的最小抑菌浓度(MIC)及最小杀菌浓度(MBC);(2)生物膜形成抑制试验测定伊犁黑蜂蜂胶对口腔常见致龋菌的最小生物膜形成抑制浓度(MBIC50);(3)通过结晶紫染色法测定伊犁黑蜂蜂胶对口腔常见致龋菌的最小生物膜清除浓度(MBEC);结果(1)伊犁黑蜂蜂胶对变形链球菌、远缘链球菌、嗜酸乳杆菌、血链球菌、粘性放线菌和内氏放线菌的最低抑菌浓度分别是0.78、0.39、1.56、0.39、0.20和0.20 mg/mL,最低杀菌浓度分别为1.56、0.78、3.125、0.78、0.39和0.39mg/mL;(2)伊犁黑蜂蜂胶对变形链球菌、远缘链球菌、血链球菌、粘性放线菌和内氏放线菌的MBIC50分别是0.39、0.39、0.39、0.05和0.10mg/mL;(3)伊犁黑蜂蜂胶对变形链球菌、远缘链球菌、血链球菌、粘性放线菌和内氏放线菌的MBEC分别是6.25、1.56、3.1256、0.78和0.78mg/mL。结论伊犁黑蜂蜂胶对口腔主要致龋细菌的生长具有抑制作用,能够抑制主要致龋细菌单菌生物膜的形成,并具有一定清除作用,是具有一定防龋效果的天然药物。     

血链球菌产生过氧化氢机理研究

作者在国内首次建立两种确定细菌产生过氧化氦的方法,可以定性和定量分析。本文探讨了血链球菌产生过氧化氢的条件,研究其在牙周微生态环境中的作用。     

Genetic exchange between oral streptococci during mixed growth

To determine whether oral streptococci might exchange genetic information in the oral cavity, paired transformable strains of Streptococcus mutans, Streptococcus sanguis and Streptococcus milleri were growth together. Chromosomal and plasmid-borne antibiotic resistance markers could be readily transferred from S. mutans GS-5 to S. milleri NCTC 10707 or S. sanguis Challis during mixed growth. However, no exchange from the latter two organisms to strain GS-5 could be detected under these conditions. The transfer of genetic information from S. sanguis to S. milleri was also observed.     

Interbacterial adhesion between Pseudomonas aeruginosa and indigenous oral bacteria isolated from patients with cystic fibrosis

Interbacterial adhesion between strains of Pseudomonas aeruginosa and strains of indigenous oral bacteria, both of which were isolated from the oral cavity of cystic fibrosis patients, was investigated by the phenomenon of the coaggregation reaction. A total of 22 strains of P. aeruginosa were isolated from the oral cavity of 17 patients and examined for their abilities to coaggregate with 5 strains each of Streptococcus sanguis, Streptococcus mitis, Actinomyces viscosus, and Actinomyces naeslundii. Coaggregation reactions were common between these oral bacteria and both the mucoid and nonmucoid variants of P. aeruginosa. All strains of P. aeruginosa were also able to agglutinate neuraminidase-treated or untreated human erythrocytes of blood types A, B, and O. Positive coaggregation reactions were further characterized by determining the effects of several sugars, and of heat and protease treatments of the bacteria. None of the coaggregtion reactions were inhibited by 0.05 M lactose, galactose, glucose, fucose, or mannose. All coaggregation reactions were dependent upon heat- and protease-sensitive components of the Pseudomonas. Thus, the interbacterial adhesions between P. aeruginosa and the oral bacteria studied appears to involve adhesins on the Pseudomonas cell, which bind to complementary receptors, on the cell surfaces of oral bacteria. The apparent prevalence and diversity of interbacterial adhesions between P. aeruginosa strains originating from the oral cavity of cystic fibrosis patients and strains of the indigenous oral bacteria suggest that some of these reactions may affect the extent to which P. aeruginosa colonizes in the oral cavity of cystic fibrosis patients, and thereby, influence susceptibility of the host to infection.     

Coaggregation properties of human oral Veillonella spp.: relationship to colonization site and oral ecology.

The primary habitats of oral veillonellae are the tongue, dental plaque, and the buccal mucosa. Isolates were obtained from each habitat and tested for coaggregation with a battery of other oral bacterial strains. All 59 tongue isolates tested for coaggregation were Veillonella atypica or Veillonella dispar. All but one of them coaggregated with strains of Streptococcus salivarius, a predominant inhabitant of the tongue surface but not subgingival dental plaque. These tongue isolates were unable to coaggregate with most normal members of the subgingival flora such as Actinomyces viscosus, Actinomyces naeslundii, Actinomyces israelii, and Streptococcus sanguis. In contrast, 24 of 29 Veillonella isolates, of which 20 were Veillonella parvula from subgingival dental plaque samples, coaggregated strongly with the three species of Actinomyces, S. sanguis, and other bacteria usually present in subgingival plaque, but they did not coaggregate with S. salivarius. The majority of isolates from the buccal mucosa (42 of 55) has coaggregation properties like those from the tongue. These results indicate that the three human oral Veillonella species are distributed on oral surfaces that are also occupied by their coaggregation partners and thus provide strong evidence that coaggregation plays a critical role in the bacterial ecology of the oral cavity.     

Coaggregation properties of human oral Veillonella spp.: relationship to colonization site and oral ecology

The primary habitats of oral veillonellae are the tongue, dental plaque, and the buccal mucosa. Isolates were obtained from each habitat and tested for coaggregation with a battery of other oral bacterial strains. All 59 tongue isolates tested for coaggregation were Veillonella atypica or Veillonella dispar. All but one of them coaggregated with strains of Streptococcus salivarius, a predominant inhabitant of the tongue surface but not subgingival dental plaque. These tongue isolates were unable to coaggregate with most normal members of the subgingival flora such as Actinomyces viscosus, Actinomyces naeslundii, Actinomyces israelii, and Streptococcus sanguis. In contrast, 24 of 29 Veillonella isolates, of which 20 were Veillonella parvula from subgingival dental plaque samples, coaggregated strongly with the three species of Actinomyces, S. sanguis, and other bacteria usually present in subgingival plaque, but they did not coaggregate with S. salivarius. The majority of isolates from the buccal mucosa (42 of 55) has coaggregation properties like those from the tongue. These results indicate that the three human oral Veillonella species are distributed on oral surfaces that are also occupied by their coaggregation partners and thus provide strong evidence that coaggregation plays a critical role in the bacterial ecology of the oral cavity.     

Salivary film expresses a complex, macromolecular binding site for Streptococcus sanguis

Teeth in the oral cavity are coated with a salivary film or pellicle, which lacks apparent intermolecular organization. This heterogeneous film facilitates binding of early commensal colonizing bacteria, including Streptococcus sanguis. To test the hypothesis that sufficient intermolecular organization exists in salivary films to form binding sites for S. sanguis, an in vitro model of saliva-coated teeth was probed with murine anti-idiotypical monoclonal antibodies (mAb2, anti-ids). The anti-ids were harvested from hybridomas that were developed in response to first generation murine hybridomas that produced anti-S. sanguis adhesin monoclonal antibodies (mAb1). The anti-ids (i) reacted with experimental salivary films and inhibited S. sanguis adhesion in a dose-dependent fashion. In Western blots, the anti-ids (ii) recognized a high molecular weight salivary antigen and (iii) secretory IgA (sIgA) light chain and alpha-amylase. After isolation by gel filtration from whole saliva or mixed secretory IgA and alpha-amylase, the high molecular weight component, containing amylase activity and sIgA, bound to hydroxyapatite to promote adhesion of S. sanguis. Therefore, a complex enriched in secretory immunoglobulin A and alpha-amylase forms a S. sanguis-binding site.     

Determination of the cell surface hydrophobicity of oral bacteria using a modified hydrocarbon adherence method

Abstract Hydrophobic interactions between bacterial cell surfaces and colonisable substrates have been implicated in the mechanisms of bacterial adherence. However, current methods of assessing bacterial hydrophobicity as a function of adherence to liquid hydrocarbons (especially hexadecane) do not always produce accurate or reproducible results. Therefore, the present technique was developed using xylene. The hydrophobic surface properties of fresh and type strains of Bacteriodes gingivalis, Bacteriodes intermedius, Capnocytophaga spp., Streptococcus salivarius and Streptococcus sanguis suspended either in saliva ions buffer (SIB) or in saliva diluted in SIB were measured. In SIB the test strains were predominantly hydrophobic. The addition of saliva caused a significant reduction ( P < 0.05) in hydrophobicity compared to SIB alone, with 80% of the strains tested. Since oral bacteria will be suspended in saliva in vivo, it is concluded that bacteria in the oral cavity may be less hydrophobic than previous studies have suggested.     

天然药物对血链球菌生长和产酸影响的体外研究

目的：通过研究不同天然药物对血链球菌生长及产酸的影响,为今后筛选出能有效调理口腔菌群生态平衡的药物奠定基础。方法：选用变形链球菌SB179作为实验菌株,测定川芎,血藤,五倍子等11种天然药物的最低抑菌浓度MIC,再以低于MIC的4个浓度配制含药的TPY液体培养基,调定其初始pH为7．4,接种血链球菌,厌氧培养后测定其终末pH,结果：当药物浓度低于或等于8．000mg/ml时,各天然药物对血链球菌的生长均有一定的抑制作用,且以五倍子作用较强,槟榔,茶多酚,蜂房,大黄,三七,血藤,白芷对血链球菌的产酸具有一定的抑制能力,而黄芩,川芎,五倍子和儿茶没有明显的抑制作用,结论：槟榔,茶多酚,蜂房,大黄,三七,血藤和白芷对血链球菌的生长和产酸都有一定的抑制作用.     

Bactericidal action of carbon dioxide laser radiation in experimental dental root canals

The ability of a carbon dioxide laser to sterilize the root canal of human teeth has been investigated. Three oral bacteria, Streptococcus sanguis, Streptococcus mutans, and Actinomyces viscosus, and three other bacteria, Bacillus cereus, Staphylococcus aureus, and Pseudomonas aeruginosa were used as experimental organisms. Exposure of cells on glass slides to laser radiation showed there was little difference in the exposure required to kill these six organisms. Complete recovery of bacteria from the root canal was initially a problem and was only achieved when bacterial manipulations and removal were carried out in rapid succession, within 5 min of inoculation. However, the geometry of the instrumented canal and the laser alignment were major factors in achieving consistent cell death of oral bacteria in the root canals. Using sets of 10 teeth, four repeated exposures of 10 W for 1 s was found to sterilize 4 or more of the teeth.