绝经前后细菌性阴道病厌氧菌群及耐药性分析

目的了解绝经前后细菌性阴道病患者标本中优势厌氧菌群,检测厌氧菌分离株对5-硝基咪唑类和林可霉素类抗生素的耐药性。方法采集165例未绝经和125例绝经阴道病患者以及84例未绝经和68例绝经正常妇女的阴道分泌物,测定其pH。采用选择性厌氧培养基分离类杆菌、梭杆菌、乳杆菌、优杆菌、消化链球菌和消化球菌,将分离的厌氧菌株鉴定至种。同时采用二倍平皿稀释法,测定塞克硝唑、替硝唑、甲硝唑、林可霉素和克林霉素对760株厌氧菌的最低抑菌浓度。结果100%细菌性阴道病患者、97.6%未绝经和98.5%绝经正常妇女的阴道分泌物中分离出厌氧菌。阴道炎患者标本中,类杆菌属、普氏菌属和紫质单胞菌属厌氧菌分离率高于相应对照组(P<0.05;P<0.01),其中以多毛类杆菌、腐败类杆菌、脆弱类杆菌、产黑普氏菌、不解糖紫质单胞菌为优势菌群;梭杆菌属、优杆菌属、消化球菌属和消化链球菌属厌氧菌分离率与相应对照组比较差异无显著性(P>0.05);乳杆菌属分离率明显低于相应对照组(P<0.01)。未绝经正常妇女和患者及绝经正常妇女和患者阴道分泌物pH分别为4.5±0.2、5.5±0.8、6.5±0.3和6.5±1.0。塞克硝唑、替硝唑和甲硝唑对355株革兰阴性厌氧菌M IC90值为1~8μg/m l,林可霉素和克林霉素则为4~16μg/m l;塞克硝唑、替硝唑和甲硝唑对405株革兰阳性厌氧菌M IC90值为8~32μg/m l,林可霉素和克林霉素则为4~16μg/m l。     

间隙感染和冠周炎标本中厌氧菌的分离鉴定和体外药效学的研究

本文报告１７０例间隙感染和２１０例冠周炎标本中厌氧菌分离鉴定及体外药效学试验的结果。８９．４％间隙感染标本中检出的厌氧菌,以产黑色素类杆菌、具核梭杆菌和衣氏放线菌较为多见。８４．３％冠周炎标本中检出的厌氧菌,以消化链球菌、韦荣球菌和具核梭杆菌较为多见。纸片扩散法药敏试验和ＭＩＣ测定结果表明替硝唑和克林霉素抗厌氧菌作用明显较甲硝唑和乙酰螺旋霉素为强（ｘ~＞１０．７４,Ｐ＜０．０１）;替硝唑与克林霉素的抗菌谱有差异,时Ｇ~-厌氧菌,替硝唑抑菌活性明显优于克林霉素,对Ｇ~＋厌氧菌,替硝唑抑菌活性不如克林霉素（ｘ~２＞２５．１９,ｐ＜０．０１）。     

牙周炎患者龈下厌氧菌群的分离和鉴定

从34例成年人牙周炎和18例青少年牙周炎患者及24例正常人的龈下菌斑中分离并鉴定了26种共311株革兰氏阴性无芽胞厌氧菌,其中产黑素类杆菌、牙龈类杆菌、伴放线嗜血杆菌、具核梭杆菌、衣氏放线菌和梅氏放线菌是牙周炎患者龈下优热菌群,小韦荣氏球菌的检出率与正常人比较无显著性差异。实验结果提示牙周炎为非单一的病原体所引     

成年牙周炎病人龈下菌斑优势菌群的探讨

本文用2种非选择性培养基和12种选择性培养基对17名北京市成年牙周炎病人和9名健康对照者龈下及龈沟菌斑菌群进行了培养、分离和鉴定。结果,牙周炎患者龈下菌斑的菌群主要由专性厌氧的革兰氏阴性杆菌组成(60.8％),优势菌为牙龈拟杆菌等;而对照组的菌群主要由兼性厌氧的革兰氏阳性球菌组成(69.2％),优势菌为链球菌属等。这种菌群构成,两组样本存在显著性差异,对于我们认识牙周炎的病原及有关微生态学问题可能有重要意义。     

牙周炎病人龈下韦荣球菌的实验观察

目的 探讨牙周炎病人龈下韦荣球菌的变化及牙周炎的细菌因素。方法 采用厌氧培养及活菌计数（CFU）的方法检测莲球菌（Streptococcus）、韦荣球菌（Veillonella）、乳杆菌(Lactobacillus）及类杆菌（Bacteroids)的含量。结果牙周炎病人龈下莲球菌含量显著低于正常对照组（P＜0.001）,韦荣球菌、乳杆菌及类杆菌的含量显著高于正常对照组（P＜0.001）。结论 牙周炎病人龈下菌群处于严重失调状态,韦荣球菌对牙周炎发病的作用应引起高度重视。     

胃癌及癌旁组织中无芽孢厌氧菌相关性的研究

在实验研究过程中发现 ,不同的消化道疾病患者 ,胃黏膜微生物群的检出率、菌种、数量均有不同 ,其中胃癌组微生物群改变最明显。为了解微生物与胃癌的相关性 ,对 5 7例胃癌患者的癌组织和癌旁组织进行了无芽孢厌氧菌的检测 ,实验结果表明 ,胃癌病变的中心部位与癌旁组织的无芽孢厌氧菌的检出情况不同。胃癌组织检出的主要为革兰氏阳性无芽孢厌氧菌 (优杆菌、丙酸杆菌、消化链球菌等 ) ,占检出菌株的 6 4 .86 %(48/74 ) ,其中硝酸盐还原实验阳性菌株为 5 9.4 6 % (44 /74 )。癌旁组织检出的主要为革兰氏阴性无芽孢厌氧菌 (类杆菌、梭杆菌、紫单胞菌、韦荣球菌 ) ,占检出细菌的 5 7.14 % (2 4 /42 ) ,硝酸盐还原实验阳性菌株为16 .6 7% (7/42 )。胃癌的病因受多种因素的影响 ,在胃癌组织中检测到无芽孢厌氧菌对胃癌的形成起到的作用 ,有待进一步研究。     

5—硝基咪唑类药物体外抗无芽孢厌氧菌效果的观察

从多种临床标本中分离并鉴定了200株无芽孢厌氧菌.赛克硝唑(SNZ)、替硝唑(TNZ)和甲硝唑(MNZ)对120株革兰氏阴性厌氧菌的MIC90分别为1～4,1～2和4～8 mg/L,对80株革兰氏阳性厌氧菌的MIC90分别为4～8,4～16和16～32 mg/L.SNZ体外抑制革兰氏阴性厌氧菌作用略弱于TNZ,但抑制革兰氏阳性无厌氧菌较TNZ稍强,MNZ对两类厌氧菌的抑制效果均不如SNZ和TNZ.     

感染根管的厌氧菌培养结果分析

从64只感染根管中的58只根管分离到144株无芽胞厌氧菌,其中类杆菌54株,厌氧性链球菌23株,韦荣氏球菌17株,真杆菌11株,梭杆菌10株,放线菌8株,双岐杆菌2株,消化链球菌和消化球菌19株。40只根管为厌氧菌和兼性厌氧菌或需氧菌混合感染,18只根管和6只根管分别为单独厌氧菌和兼性厌氧菌感染。33只根尖周炎根管分别采集牙髓和根尖渗出物样本进行培养,实验结果表明牙髓样本中革兰氏阳性厌氧杆菌检出率较高,根尖渗出物中以产黑素类杆菌属的细菌检出率较高。根尖周炎和牙槽脓肿患者的感染根管中产黑素类杆菌属的细菌检出率明显高于蜂窝组织炎患者。     

替硝唑治疗冠周炎,牙周炎的临床及微生态学评价

应用替硝唑片口服治疗４０例冠周炎、牙周炎患者,以甲硝唑为对照检测了治疗前后冠周盲袋及牙周袋内厌氧菌的变化。结果表明,服用替硝唑后,袋内厌氧菌的数量及种类明显下降,而球菌的数量明显增多。替硝唑治疗冠周炎、牙周炎的疗效（９２．５％）优于甲硝唑（６５．０％）。替硝唑对冠周袋及牙周袋的厌氧菌杀灭强度及速度较甲硝唑好,特别是对产黑色素Ｇ－厌氧杆菌,不产黑色素Ｇ－厌氧杆菌,小韦荣氏菌,ＣＯ２噬纤维菌具有很强的杀灭效果。     

儿童乳牙根尖周炎感染根管厌氧细菌学分析

目的：报告50例5-8.5岁儿童根尖周炎患者感染根管73颗乳牙厌氧菌分离鉴定及体外药敏试验的结果,方法：采用K-B法。结果：89.04%(65/73)的患牙共检出厌氧菌150株,平均每例标本检出2.3株,常用抗厌氧菌药物替硝唑（TNZ）、甲硝唑（MNZ）对革兰阴性厌氧菌（产黑色素普氏菌,中间普氏菌,解脲拟杆菌,具核梭杆菌,牙龈二氧化碳噬纤维菌）有较好的抑菌作用;林可霉素对革兰阳性厌氧菌（厌氧消化链球菌,嗜酸乳杆菌）的作用则强于TNZ和MNZ,氯霉素（C）和乙旋螺旋霉素（AS）抗厌氧菌的作用则弱于前三者。结论：分析儿童乳牙根尖周炎感染根管菌群来决定抗菌药物的选择,在临床治疗上是必要的。     

Microbiological and clinical effects of a 1% chlorhexidine-gel in untreated periodontal pockets from adult periodontitis patients.

The aim of this study was to report the microbiological and clinical effects of repeated subgingival administration of a 1% Chlorhexidine-gel in periodontal pockets from 10 patients with adult periodontitis. Results showed that the experimental treatment significantly improved clinical parameters (Plaque Index, Gingival Bleeding Index, and Pocket Probing Depth). Direct subgingival administration of Chlorhexidine-gel also produced a remarkable modification in the proportions of putative periodontopathic microorganisms, such as Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Veillonella parvula, Fusobacterium nucleatum, and Peptostreptococcus micros, in subgingival bacterial plaque from periodontitis patients.     

In vitro antimicrobial activity of propolis samples from different geographical origins against certain oral pathogens

Propolis is an agent having antimicrobial properties, however, its composition can vary depending on the area where it is collected. In the present study, the antimicrobial activity of five propolis samples, collected from four different regions in Turkey and from Brazil, against nine anaerobic strains was evaluated. Ethanol extracts of propolis (EEP) were prepared from propolis samples and we determined minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of EEP on the growth of test microorganisms by using agar dilution method. All strains were susceptible and MIC values ranged from 4 to 512 microg/ml for propolis activity. Propolis from Kazan-Ankara showed most effective MIC values to the studied microorganisms. MBC values of Kazan-Ankara EEP samples were ranged from 8 to 512 microg/ml. Death was observed within 4 h of incubation for Peptostreptococcus anaerobius and micros and Lactobacillus acidophilus and Actinomyces naeslundii, while 8 h for Prevotella oralis and Prevotella melaninogenica and Porphyromonas gingivalis, 12 h for Fusobacterium nucleatum, 16 h for Veillonella parvula. It was shown that propolis samples were more effective against Gram positive anaerobic bacteria than Gram negative ones. The organic chemical compositions of EEPs were determined by high-resolution gas chromatography coupled to mass spectrometry (GC-MS). The main compounds of EEPs were flavonoids such as pinobanksin, quercetin, naringenin, galangine, chrysin and aromatic acids such as cafeic acid. Because of increased antimicrobial resistance, propolis may be kept in mind in the treatment of oral cavity diseases.     

Quantification of Subgingival Bacterial Pathogens at Different Stages of Periodontal Diseases

Anaerobic gram-negative oral bacteria such as Treponema denticola, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Campylobacter rectus, and Fusobacterium nucleatum are closely associated with periodontal diseases. We measured the relative population (bacterial levels) of these oral pathogens in subgingival tissues of patients at different stages of Korean chronic periodontal diseases. We divided the individuals into those with chronic gingivitis (G), moderate periodontitis (P1), severe periodontitis (P2), and normal individuals (N) (n?=?20 for each group) and subgingival tissue samples were collected. We used real-time PCR with TaqMan probes to evaluate the change of periodontal pathogens among different stages of periodontitis. Bacterial levels of A. actinomycetemcomitans and C. rectus are significantly increased in individuals with chronic gingivitis and moderate periodontitis, but unchanged in severe periodontitis patients. These results suggest that analyzing certain bacterial levels among total oral pathogens may facilitate understanding of the role of periodontal bacteria in the early stages of periodontitis.     

Denaturing gradient gel electrophoresis analysis with different primers of subgingival bacterial communities under mechanical debridement

DGGE of 16S rDNA is one of the most frequently used methods to study microbial communities. In this study, the DGGE profiles of different 16S rDNA regions of the periodontal pathogens Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella nigrescens were investigated. The results suggested that V3-V5 and V6-V8 fragments may be suitable for community analysis of subgingival bacteria. Further analysis of subgingival samples with V3-V5 and V6-V8 regions as target fragments suggested that, in chronic periodontitis, re-colonization by periodontal bacteria with a population very similar to the baseline may occur by 6 weeks after mechanical debridement.     

Comparison of culture methods and multiplex PCR for the detection of periodontopathogenic bacteria in biofilm associated with severe forms of periodontitis

Conventional culture methods and Multiplex PCR, both of which we have been used for a long time in our clinical microbiology laboratory, were compared for their ability to detect a selected panel of periodontopathic bacteria: Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, and Prevotella intermedia. Tests were performed in a single subgingival sample taken from a periodontal diseased site with a probing depth equal to or greater than 6mm. The results were compared site-by-site, taking into account the quality and the presence or absence of pathogens. 529 samples of subgingival plaque were analysed and the prevalence of the six species monitored varied in relation to the species itself and the method of detection. The most represented species is F. nucleatum, with a percentage of positive variability between 44.9% PCR and 46.5% culture test. Generally, the lowest prevalence was determined by culture test, with the exception of E. corrodens and F. nucleatum, which, unlike other bacteria, have been seen in higher percentages in culture with respect to PCR. For both methods, there was a good degree of accuracy in the determination of A. actinomycetemcomitans, C. rectus, E. corrodens, and P. gingivalis. It becomes weak for F. nucleatum and P. intermedia. Both culture and PCR techniques introduced many methodological problems when applied in oral microbiology, but the ideal technique for accurate detection of pathogens in subgingival plaque samples has yet to be developed.     

Identification and localization of extraradicular biofilm-forming bacteria associated with refractory endodontic pathogens

Bacterial biofilms have been found to develop on root surfaces outside the apical foramen and be associated with refractory periapical periodontitis. However, it is unknown which bacterial species form extraradicular biofilms. The present study aimed to investigate the identity and localization of bacteria in human extraradicular biofilms. Twenty extraradicular biofilms, used to identify bacteria using a PCR-based 16S rRNA gene assay, and seven root-tips, used to observe immunohistochemical localization of three selected bacterial species, were taken from 27 patients with refractory periapical periodontitis. Bacterial DNA was detected from 14 of the 20 samples, and 113 bacterial species were isolated. Fusobacterium nucleatum (14 of 14), Porphyromonas gingivalis (12 of 14), and Tannellera forsythensis (8 of 14) were frequently detected. Unidentified and uncultured bacterial DNA was also detected in 11 of the 14 samples in which DNA was detected. In the biofilms, P. gingivalis was immunohistochemically detected in all parts of the extraradicular biofilms. Positive reactions to anti-F. nucleatum and anti-T. forsythensis sera were found at specific portions of the biofilm. These findings suggested that P. gingivalis, T. forsythensis, and F. nucleatum were associated with extraradicular biofilm formation and refractory periapical periodontitis.     

Serum antibodies of periodontitis patients compared to the lipopolysaccharides of Porphyromonas gingivalis and Fusobacterium nucleatum

Serum antibody titers against the lipopolysaccharides (LPSs) of Porphyromonas gingivalis and Fusobacterium nucleatum were compared between 9 periodontitis patients and 24 healthy persons. The IgG titers against the LPSs of P. gingivalis ATCC 33277(T) and W50 were clearly higher in the patients than in the healthy persons. However, IgM titers against the LPSs of P. gingivalis strains were relatively low, and no significant difference was observed between the patients and healthy persons. On the other hand, IgG and IgM titers against the LPS of Fusobacterium nucleatum JCM 8532(T) in some patients were significantly higher than those in the healthy persons, although the difference in IgG titers was not large compared to that of the LPS of P. gingivalis. These results suggest that the antibody measurement of patients' sera against the LPS of periodontal bacteria can be applied for the diagnosis of periodontitis.     

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。     

Periodontitis in established rheumatoid arthritis patients: a cross-sectional clinical,microbiological and serological study

Introduction

The association between rheumatoid arthritis (RA) and periodontitis is suggested to be linked to the periodontal pathogen Porphyromonas gingivalis. Colonization of P. gingivalis in the oral cavity of RA patients has been scarcely considered. To further explore whether the association between periodontitis and RA is dependent on P. gingivalis, we compared host immune responses in RA patients with and without periodontitis in relation to presence of cultivable P. gingivalis in subgingival plaque.

Methods

In 95 RA patients, the periodontal condition was examined using the Dutch Periodontal Screening Index for treatment needs. Subgingival plaque samples were tested for presence of P. gingivalis by anaerobic culture technique. IgA, IgG and IgM antibody titers to P. gingivalis were measured by ELISA. Serum and subgingival plaque measures were compared to a matched control group of non-RA subjects.

Results

A higher prevalence of severe periodontitis was observed in RA patients in comparison to matched non-RA controls (27% versus 12%, p < 0.001). RA patients with severe periodontitis had higher DAS28 scores than RA patients with no or moderate periodontitis (p < 0.001), while no differences were seen in IgM-RF or ACPA reactivity. Furthermore, RA patients with severe periodontitis had higher IgG- and IgM-anti P. gingivalis titers than non-RA controls with severe periodontitis (p < 0.01 resp. p < 0.05), although subgingival occurrence of P. gingivalis was not different.

Conclusions

Severity of periodontitis is related to severity of RA. RA patients with severe periodontitis have a more robust antibody response against P. gingivalis than non-RA controls, but not all RA patients have cultivable P. gingivalis.     

三种根管充填材料对 乳牙根尖周炎常见细菌的抑菌性

目的评估和比较3种乳牙根管充填材料对乳牙根尖周炎常见细菌的抗菌功效。方法在体外实验中,采用纸片扩散法检测并观察3种乳牙根管充填材料及蒸馏水作用于粪肠球菌、牙龈卟啉单胞菌和具核梭杆菌后形成的抑菌环直径(mm),用SPSS 24.0软件进行统计分析。结果三重抗生素糊剂对乳牙根尖周炎根管中3种细菌均显示出最强的抗菌潜力,其次是Vitapex?、氢氧化钙糊剂,结果差异具有统计学意义(P0.05);蒸馏水对3种细菌是非抑制性的。粪肠球菌对3种药物反应最敏感,其次是牙龈卟啉单胞菌、具核梭杆菌,结果差异具有统计学意义(P0.05)。结论三重抗生素糊剂是最推荐用于治疗乳牙根尖周炎的根管充填材料。