刚果红负性染色法检查口腔细菌

Listgarten 首先报道用暗视野检查法测定牙周细菌的百分组成比,以了解口腔牙周炎患者龈下菌群的组成比与不同临床状态的关系。由于暗视野法计数较难,并要求配备暗视野显微镜而不易推广。笔者近年来用刚果红负性染色法代替暗视野法检测口腔菌斑、唾液、拔牙创干槽症分泌物等临床标本的细菌组成百分比取得了理想的结果,与暗视野法比较,该法除不能检测可动菌外,具有不需特殊的仪器设备,操作方便,容易观察、快速、准确、试剂容易获得、配制简单、细菌涂片可保存等优点。     

固定矫治器对口腔菌群的影响

目的利用PCR-DGGE方法探讨戴用牙齿固定矫治器对口腔菌群的影响。方法从20例健康的牙齿正常个体和30例戴用固定矫治器的正畸患者的唾液中提取细菌基因组总DNA,利用聚合酶链式反应—变形梯度凝胶电泳(PCR-DGGE)方法获取口腔菌群的分子指纹图谱,对其进行相似性和多样性的分析以及优势条带的序列分析。结果 PCR-DGGE可明确将正常个体和正畸患者分成2个簇,且正畸患者口腔菌群的多样性明显增加,优势菌群发生转变,序列分析表明口腔链球菌数量明显减少,假单胞菌成为优势菌型。结论戴用固定矫治器能改变口腔菌群分布,引起口腔中有益菌的减少,促使致病菌增长,导致口腔菌群失调。     

拔牙术前后类杆菌及梭杆菌变化的临床意义

拔牙术破坏了口腔的正常生理结构,改变了口腔内环境,使菌群失调。未实验对１７名拔牙的患者进行了拔牙术前术后类杆菌、梭杆菌的培养,结果表明,无论是类杆菌还是梭杆菌在拔牙前后都发生了明显的变化,尤其是梭杆菌拔牙后明显高于拔牙前。     

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。     

2型糖尿病小鼠食道菌群的结构分析

人体内庞大的微生物群体对人体有着巨大的影响.越来越多的数据表明,肠道菌群与肥胖症、糖尿病等代谢性疾病的发生密切相关.食道菌群结构对研究胃肠道及整个消化 系统菌群结构至关重要.针对10只2型糖尿病模型小鼠及10只正常对照小鼠食道样本,进行变性梯度凝胶电泳（DGGE）和克隆测序分析食道菌群结构.结果发现,实验组小鼠与对照组小鼠食道菌群多样性指数与丰富度指数存在显著差异,均匀度指数无显著差 异.说明正常小鼠较2型糖尿病小鼠食道菌群种类及数量较大,优势菌种类及相对含量相似.测序结果显示,正常小鼠食道内含乳杆菌属细菌,而患病小鼠食道内不含乳杆菌属细菌或含量极低.提示乳杆菌属细菌与2型糖尿病密切相关,实验结果对研究糖尿病发病机理及并发症治疗有重要意义.     

口腔菌群中螺旋体的分布

螺旋体是口腔正常菌群的一个组成部分.长期以来,一直被认为与牙周疾病的发生关系密切.在坏死性龈炎病变部位,可以发现大量的螺旋体,在慢性牙周炎的病损区也可发现菌斑中螺旋体的百分     

中药透皮吸收抗菌活性的测试及对实验动物口腔菌群的影响

通过对黄连解毒透皮吸收剂经豚鼠背部透皮吸收后的血清药物浓度测定、体外抗菌活性测定、动物口腔菌群变化结果研究其抗感染的生物学效应及对正常菌群的影响。结果表明受试动物血清中可检测到有效成分并显示出抗菌活性,同时可引起口腔细菌菌群的变化。     

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.     

口腔鳞状细胞癌患者术后病变区菌群分析

摘要：目的 研究口腔鳞状细胞癌患者术后病变区菌群变化。方法 选择2017年12月至2018年12月于我院进行治疗的80例口腔鳞状细胞癌患者为研究对象,测定入选患者术前和术后第14天口腔病变中心区细菌检出率、构成比和数量。结果 患者术前病变区链球菌属的构成比（67.63%）和检出率（93.89%）最高,其次为葡萄球菌属、奈瑟菌属和普氏菌属。检出细菌数量为（4.06±0.42）~（5.49±0.53）CFU/cm2。患者术后病变区链球菌属的构成比（75.48%）和检出率（81.45%）最高,其次为普氏菌属、韦荣菌属、葡萄球菌属。检出细菌数量为（2.76±0.39）~（4.54±0.63）CFU/cm2。患者手术前后奈瑟菌属和梭杆菌属的检出率和数量差异有统计学意义（P<0.05）,二氧化碳噬纤维菌属数量和细菌总量差异有统计学意义（P<0.05）,而病变区各细菌构成比差异无统计学意义（P>0.05）。结论 口腔鳞状细胞癌患者术后链球菌属的构成比和检出率最高,未检测出梭杆菌属和奈瑟菌属细菌。     

感受态与口腔链球菌多种表型间关系的研究进展

链球菌是人类口腔中最为常见的细菌类群之一,在口腔微生态平衡的维持与致病中发挥了重要作用。口腔链球菌中的大多数可以进入感受态,在此生理状态下,细菌可摄取环境中的DNA并整合进入自身基因组从而获得新的遗传表型或特性。大量研究表明,口腔链球菌的感受态调控通路不是孤立的,与生物膜形成、细菌素产生、耐酸、氧应激、细胞自溶和耐药性等多个表型的调控存在紧密关系,研究这些不同表型间的相互影响对理解口腔菌群稳态及防治疾病有重要意义。本文以变异链球菌、格氏链球菌、血链球菌和肺炎链球菌4种典型的口腔链球菌为代表,对感受态与口腔链球菌多种表型间关系的研究进展做一综述。     

慢性根尖周炎的细菌学研究

本文对14例慢性根尖周炎患者16个感染根管细菌培养检查的结果发现:以厌氧菌为优势的混合菌感染是慢性根尖周炎感染根管细菌学的主要特点。穿髓根管由于敞开的髓腔,同一根管内4—9种细菌混合感染者占87.5％;未穿髓的感染根管2—3种细菌混合感染的占62.5％,4种以上细菌混合感染的占37.5％。优势的感染菌包括产黑色素类杆菌、口腔类杆菌、口类杆菌、具核梭杆菌、干酪乳杆菌、内氏放线菌等6种厌氧菌;血液链球菌、变链球菌和酿脓链球菌为最常见的兼性厌氧菌。本文指出口腔正常菌群是感染根管的机会病原菌。     

需氧菌阴道炎、细菌性阴道病阴道菌群分析

用阴道分泌物直接涂片、革兰染色、选择性培养分离和细菌预成酶谱分析等微生物学方法和生物化学方法,分析了正常女性、AV、BV患者的阴道菌群。正常女性阴道中以乳酸杆菌为主,产H2O2乳酸杆菌的检出率89%,AV的致病菌主要是革兰阳性率需氧菌、检出率可达80%,其中金黄色葡萄球菌,粪肠球菌和埃希氏大肠菌较为常见,B族链球菌的检出率较低;BV的致病菌主要是厌氧菌,以革兰阴性厌氧菌的检出率最高,加德纳菌的检出率不足40%。48例BV患者中有8例合并AV感染,31例AV患者中有6例合并BV感染。细菌谱分析发现,AV患者易感染消化链球菌、普雷沃菌等厌氧菌和加德纳菌,BV患者易感染粪肠球菌、大肠埃希菌等需氧菌。     

The presence of bacteria and yeast like fungi in specimens from surgical patients

The aim of this study was to determine the incidence of Candida spp. strains in specimens obtained from surgically treated patients as well as to analyze the accompanying bacterial flora, both aerobic and anaerobic. The material came from two groups of patients. In the first group consisting of patients operated for colon and rectum carcinoma, the samples included peritoneal fluid, colon or rectum bioptates, pus, blood, and wound swabs. In the other group, biopsy material and smears from post operation wounds were taken from patients who underwent a surgical treatment of larynx carcinoma. Altogether, 282 various clinical specimens from 165 patients were analysed, and 41 Candida spp. strains were isolated: 39 strains of C. albicans and 2 strains of C. tropicalis. In 20 out of 41 specimens infected with Candida spp. (48.8%) the co-infection with bacterial aerobic flora was found. In 10 cases (24.4%), the fungi were isolated together with aerobic and anaerobic bacterial flora, whereas in 2 specimens (4.9%) the anaerobes and Candida albicans were diagnosed. The remaining 9 samples showed only the presence of Candida spp. (21.9%). From among aerobic bacterial flora Enterococcus spp. strains (n = 17) and Gram negative rods from Enterobacteriaceae family (n = 13) were the most frequently isolated. The bacterial strains of Streptococcus spp. (n = 5), Pseudomonas spp. (n = 3), Staphylococcus spp. and Corynebacterium spp. (2 strains, both) were identified more rarely. Bacteroides spp. were the most frequent members of bacterial anaerobic flora (n = 10). Other isolated anaerobic bacteria were classified as Fusobacterium spp. or Peptostreptococcus spp. (1 strain each). E. coli and Enterococcus spp. strains of aerobic bacterial flora were more frequently isolated together with Candida spp. CONCLUSIONS: (i) Mixed bacterial flora was found to predominate in the clinical material from the patients after surgery. (ii) Candida spp. were most frequently found together with aerobic bacterial flora.     

Bacteroides and appendicitis (author's transl)]

Bacteroides fragilis is frequently recovered from cases of appendicitis with perforation and from infections developing secondary to appendicitis. In order to assess the part played by B. fragilis in the aetiology of appendicitis, quantitative aerobic and anaerobic culture studies of the contents of 49 inflammated appendices were performed. Anaerobic gram-negative non-sporing rods were cultivated from 43 appendices in the range 10(3)-10(9)/g. A total of 1,473 isolates was differentiated by biochemical methods, and 1,374 cultures were found to belong to the saccharolytic species of the genus Bacteroides (B. fragilis, B. thetaiotaomicron, B. vulgatus, B. distasonis etc.). B fragilis was detected in 31 appendices; the species predominated in 18 samples. B theraiotamicron, recovered from 27 samples, was prevalent in 4 appendices. In one sample, B. fragilis and B. thetaiotaomicron outnumbered the other appendicular bacterial. B. vulgatus was cultivated from 12 appendices, but did once constitute the prevalent group. It has been previously shown that B. vulgatus (43% of intestinal isolates) and B. thetaiomicron predominate in the normal narge bowel flora. On the other hand, approximately 80% of pyrogenic Bacteroides strains belong to B. fragilis, B. thetaiotaomicron accounting for 19% and B. vulgatus being virtually absent. From these striking differences in species distribution the conclusion was drawn that B. fragilis possesses the highest virulence for man. Species distribution within the 1,374 appendicular isolates of saccharolytic Bacteroides (percentages of 62, 19 and 4.3 for B. fragilis, B. thetaiotaomicron, and B. vulgatus, respectively) was very similar to that encountered in clinical specimens. From the results obtained it becomes evident that pyrogenic Bacteroides, in particular B. fragilis, plays an important role in nearly 50% of cases of appendicitis.     

A pilot study: microbiological conditions of the oral cavity in minipigs for peri-implantitis models

As peri-implantitis is an emerging problem, the development of validated animal models is mandatory. The aim of this pilot study was to provide a first step in describing the normal oral flora of minipigs. In five minipigs, samples of the oral flora were collected with sterile cotton swabs from the buccal gingiva of the lower jaw. Two swabs per animal were collected, followed by bacterial isolation under both aerobe and anaerobe conditions. Microbiological analyses included biochemical tests, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rDNA gene sequence analysis. A total of 61 taxa were detected, 14-21 different bacterial taxa from each minipig. Among the Gram-positive cocci, mainly staphylococcal and streptococcal species were identified. Different Actinomyces species were the most abundant taxa in the group of Gram-positive rods. Among the anaerobic bacteria, the Gram-negative genera Fusobacterium, Bacteroides and Prevotella were the most often observed taxa. This is the first study which begins to describe the normal oral flora in minipigs in cultures to allow for the detection of a broad spectrum. Several bacterial species identified are different from human ones. No specific species for peri-implantitis could be detected in that healthy sample.     

The place of molecular genetic methods in the diagnostics of human pathogenic anaerobic bacteria. A minireview

Anaerobic infections are common and can cause diseases associated with severe morbidity, but are easily overlooked in clinical settings. Both the relatively small number of infections due to exogenous anaerobes and the much larger number of infections involving anaerobic species that are originally members of the normal flora, may lead to a life-threatening situation unless appropriate treatment is instituted. Special laboratory procedures are needed for the isolation, identification and susceptibility testing of this diverse group of bacteria. Since many anaerobes grow more slowly than the facultative or aerobic bacteria, and particularly since clinical specimens yielding anaerobic bacteria commonly contain several organisms and often very complex mixtures of aerobic and anaerobic bacteria, considerable time may elapse before the laboratory is able to provide a final report. Species definition based on phenotypic features is often time-consuming and is not always easy to carry out. Molecular genetic methods may help in the everyday clinical microbiological practice in laboratories dealing with the diagnostics of anaerobic infections. Methods have been introduced for species diagnostics, such as 16S rRNA PCR-RFLP profile determination, which can help to distinguish species of Bacteroides, Prevotella, Actinomyces, etc. that are otherwise difficult to differentiate. The use of DNA-DNA hybridization and the sequencing of special regions of the 16S rRNA have revealed fundamental taxonomic changes among anaerobic bacteria. Some anaerobic bacteria are extremely slow growing or not cultivatable at all. To detect them in special infections involving flora changes due to oral malignancy or periodontitis, for instance, a PCR-based hybridization technique is used. Molecular methods have demonstrated the spread of specific resistance genes among the most important anaerobic bacteria, the members of the Bacteroides genus. Their detection and investigation of the IS elements involved in their expression may facilitate following of the spread of antibiotic resistance among anaerobic bacteria involved in infections and in the normal flora members. Molecular methods (a search for toxin genes and ribotyping) may promote a better understanding of the pathogenic features of some anaerobic infections, such as the nosocomial diarrhoea caused by C. difficile and its spread in the hospital environment and the community. The investigation of toxin production at a molecular level helps in the detection of new toxin types. This mini-review surveys some of the results obtained by our group and others using molecular genetic methods in anaerobic diagnostics.     

Aerobic growth and toxigenicity ofClostridium botulinum types A and B

Clostridium botulinum types A and B cultured in association with avian skin flora, had similar growth patterns under both aerobic and anaerobic conditions. The selective “C. botulinum isolation” (CBI) medium was found to be especially useful for the recovery and quantitation of small numbers of type A or type B organisms from the mixed cultures. Enzyme immunoassay in conjunction with conventional mouse biossay provided a practical means for the quantitation of toxigenicity ofC. botulinum in avian skin cultures. The amount of toxin produced by type A was always higher than that produced by type B strains. The aerobically incubated type A or type B cultures appeared to be less toxigenic than cultures incubated anaerobically.     

The effect of selected factors on the course of wound healing after surgery for laryngeal neoplasms

Finding of the a etiologic factors and participation of bacteria flora in wound healing in laryngeal cancer treatment was the purpose of our study. Investigations were performed in 27 patients. Swabs were taken from the postoperative wounds. Detailed identifications of the bacteria flora and antibiotic susceptibility of bacteria were performed. Wound healing was estimated according to extension of the carcinoma, applied antibiotics, state of the oral cavity, the kind of bacteriological flora isolated after surgical treatment from postoperative wounds. It was found that wound healing depended on the extension of carcinoma, as well as, type of isolated bacteria and antibiotic therapy used. The proper healing of postoperative wounds was not dependent on the state of the oral cavity and the dentition. The main cause of postoperative complication of wounds was methicillin-resistant Staphylococcus aureus (MRSA).     

Characterization of bacterial pectinolytic strains involved in the water retting process

Pectinolytic microorganisms involved in the water retting process were characterized. Cultivable mesophilic anaerobic and aerobic bacteria were isolated from unretted and water-retted material. A total of 104 anaerobic and 23 aerobic pectinolytic strains were identified. Polygalacturonase activity was measured in the supernatant of cell cultures; 24 anaerobic and nine aerobic isolates showed an enzymatic activity higher than the reference strains Clostridium felsineum and Bacillus subtilis respectively. We performed the first genotypic characterization of the retting microflora by a 16S amplified ribosomal DNA restriction analysis (ARDRA). Anaerobic isolates were divided into five different groups, and the aerobic isolates were clustered into three groups. 84.6% of the anaerobic and 82.6% of the aerobic isolates consisted of two main haplotypes. Partial 16S rRNA gene sequences were determined for 12 strains, representative of each haplotype. All anaerobic strains were assigned to the Clostridium genus, whereas the aerobic isolates were assigned to either the Bacillus or the Paenibacillus genus. Anaerobic isolates with high polygalacturonase (PG) activity belong to two clearly distinct phylogenetic clusters related to C. acetobutylicum-C. felsineum and C. saccharobutylicum species. Aerobic isolates with high PG activity belong to two clearly distinct phylogenetic clusters related to B. subtilisT and B. pumilusT.     

Association of germfree mice with intestinal microfloras obtained from "normal" mice

A cultured microflora obtained from the caecum of a "normal" mouse was given to 4 groups of germfree mice and was supplied 1x, 2x, 3x and 4x respectively at 5-day intervals. Another group received a 10(-7) dilution of the caecal flora while a group associated with an 'SPF' flora served as control. The difference (measured by 8 parameters) between mice supplied with the cultured flora or with a 10(-7) dilution, both given once only, was small. Supplying the flora 3x resulted in more 'normal' mice compared with mice which received the flora once or twice. The caeca of specified-pathogen-free mice contained more bacteria per gram (microscopic bacterial count), less aerobic and anaerobic bacteria per gram (viable counts), while the yield as percentage of the microscopic bacterial count was lower as compared with the group to which a cultured flora was supplied 4 times.