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DNA-DNA Hybridization (dna-dna + hybridization)

Distribution by Scientific Domains
Medical Sciences88%

Kinds of DNA-DNA Hybridization

  • checkerboard dna-dna hybridization
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    Selected Abstracts

    Clinical Studies Of Acinetobacter Classified By DNA-DNA Hybridization

    APMIS, Issue 5 2007
    Article first published online: 11 MAY 200
    First page of article [source]

    Bacteria of asymptomatic periradicular endodontic lesions identified by DNA-DNA hybridization

    DENTAL TRAUMATOLOGY, Issue 5 2000
    J. J. Gatti
    Abstract , Possible inclusion of contaminant bacteria during surgery has been problematic in studies of periradicular lesions of endodontic origin. Therefore, in this study, two different surgical techniques were compared. A second problem is that some difficult to cultivate species may not be detected using bacteriological methods. Molecular techniques may resolve this problem. DNA-DNA hybridization technology has the additional advantage that DNA is not amplified. The purpose of this investigation was to determine if bacteria from periradicular endodontic lesions could be identified using DNA-DNA hybridization. A full thickness intrasulcular mucoperiosteal (IS) flap (n=20) or a submarginal (SM) flap (n=16) was reflected in patients with asymptomatic apical periodontitis. DNA was extracted and incubated with 40 digoxigenin-labeled whole genomic probes. Bacterial DNA was detected in all 36 lesions. Seven probes were negative for all lesions. In patients with sinus tract communication, in teeth lacking intact full coverage crowns, and in patients with a history of trauma, 4,13 probes provided positive signals. Seven probes were positive in lesions obtained by the IS, but not the SM technique. Two probes were in samples obtained with the SM technique, but not the IS. Only Bacteroides forsythus and Actinomyces naeslundii genospecies 2 were present in large numbers using either the IS or the SM technique. The SM flap technique, in combination with DNA-DNA hybridization, appeared to provide excellent data pertaining to periradicular bacteria. These results supported other studies that provide evidence of a bacterial presence and persistence in periradicular lesions. [source]

    Change in subgingival microbial profiles in adult periodontitis subjects receiving either systemically-administered amoxicillin or metronidazole

    JOURNAL OF CLINICAL PERIODONTOLOGY, Issue 7 2001
    M. Feres
    Abstract Aim: The current investigation evaluated changes in levels and proportions of 40 bacterial species in subgingival plaque samples during, immediately after and up to 1 year after metronidazole or amoxicillin therapy combined with SRP. Method: After baseline clinical and microbiological monitoring, 17 adult periodontitis subjects received full mouth SRP and 14 days systemic administration of either metronidazole (250 mg, TID, n=8) or amoxicillin (500 mg, TID, n=9). Clinical measurements including % of sites with plaque, gingival redness, bleeding on probing and suppuration, pocket depth (PD) and attachment level (AL) were made at baseline, 90, 180 and 360 days. Subgingival plaque samples were taken from the mesial surface of all teeth in each subject at baseline, 90, 180 and 360 days and from 2 randomly selected posterior teeth at 3, 7, and 14 days during and after antibiotic administration. Counts of 40 subgingival species were determined using checkerboard DNA-DNA hybridization. Significance of differences over time was determined using the Quade test and between groups using ANCOVA. Results: Mean PD was reduced from 3.22±0.12 at baseline to 2.81±0.16 (p<0.01) at 360 days and from 3.38±0.23 mm to 2.80±0.14 mm (p<0.01) in the amoxicillin and metronidazole treated subjects respectively. Corresponding values for mean AL were 3.21±0.30 to 2.76±0.32 (p<0.05) and 3.23±0.28 mm to 2.94±0.23 mm (p<0.01). Levels and proportions of Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola were markedly reduced during antibiotic administration and were lower than baseline levels at 360 days. Counts (×105, ±SEM) of B. forsythus fell from baseline levels of 0.66±0.16 to 0.04±0.02, 0.13±0.04, 0.10±0.03 and 0.42±0.19 in the amoxicillin group at 14, 90, 180 and 360 days respectively (p<0.001). Corresponding values for metronidazole treated subjects were: 1.69±0.28 to 0.02±0.01, 0.20±0.08, 0.22±0.06 and 0.22±0.08 (p<0.001). Counts of Campylobacter species, Eubacterium nodatum, Fusobacterium nucleatum subspecies, F. periodonticum and Prevotella nigrescens were also detected at lower mean levels during and immediately after therapy, but gradually increased after withdrawal of the antibiotics. Members of the genera Actinomyces, Streptococcus and Capnocytophaga were minimally affected by metronidazole. However, amoxicillin decreased the counts and proportions of Actinomyces species during and after therapy. Conclusions: The data suggest that metronidazole and amoxicillin are useful in rapidly lowering counts of putative periodontal pathogens, but must be accompanied by other procedures to bring about periodontal stability. Zusammenfassung Ziel: Die gegenwärtige Untersuchung evaluiert die Veränderungen in den Niveaus und Proportionen von 40 bakteriellen Spezies in subgingivalen Plaqueproben während, sofort nach und bis zu 1 Jahr nach Metronidazol- oder Amoxicillintherapie in Kombination mit SRP. Methoden: Nach der klinischen und mikrobiologischen Basisuntersuchung erhielten 17 erwachsene Personen mit Parodontitis eine vollständige SRP und 14 Tage eine systemische Gabe von entweder Metronidazol (250 mg, TID, n=8) oder Amoxicillin (500 mg, TID, n=9). Die klinischen Messungen schlossen die Prozentwerte der Flächen mit Plaque, der gingivalen Rötung, der Provokationsblutung und Suppuration, der Sondierungstiefe (PD) und des Stützgewebeniveaus (AL) ein. Die Messungen wurden zur Basis, am 90., am 180. und 360. Tag gemacht. Die subgingivalen Plaqueproben wurden von der mesialen Oberfläche aller Zähne zur Basis, zum 90., zum 180. und 360. Tag von jedem Probanden genommen sowie von 2 zufällig ausgesuchten posterioren Zähnen am Tag 3, 7 und 14 während und nach der Antibiotikaverordnung. Die Mengen von 40 subgingivalen Spezies wurden unter Nutzung einer checkerboard DNA-DNA Hybridisation bestimmt. Die Signifikanzen der Differenzen über die Zeit wurden mit dem Quade-Test und zwischen den Gruppen mit der ANCOVA überprüft. Ergebnisse: Die mittleren PD reduzierten sich von 3.22±0.12 mm zur Basis zu 2.81±0.16 mm (p<0.01) zum 360. Tag und von 3.38±0.23 mm zu 2.80±0.14 mm (p<0.01) bei den mit Amoxicillin bzw. mit Metronidazol behandelten Patienten. Korrespondierende Werte für die mittleren AL waren 3.21±0.30 zu 2.76±0.32 (p<0.05) und 3.23±0.28 mm zu 2.94±0.23 mm (p<0.01). Die Niveaus und die Verteilung von Bacteroides forsythus, Porphyromonas gingivalis und Treponema denticola wurden während der Antibiotikabehandlung deutlich reduziert und waren am 360. Tag niedriger als zur Basis. Die Mengen (×105, ±SEM) von B. forsythus fielen von der Basis von 0.66±0.16 auf 0.04±0.02, 0.13±0.04, 0.10±0.03 und 0.42±0.19 in der Amoxicillin Gruppe an den Tagen 14, 90, 180 und 360 (p<0.001). Korrespondierende Werte für die mit Metronidazol behandelten Personen waren: 1.69±0.28 zu 0.02±0.01, 0.20±0.08, 0.22±0.06 und 0.22±0.08 (p<0.001). Die Mengen von Campylobacter sp., Eubacterium nodatum, Fusobacterium nucleatum subspecies, F. peridonticum und Prevotella nigrescens waren in den mittleren Niveaus während und sofort nach der Therapie auch niedriger, aber graduell erhöht nach Absetzen der Antibiotika. Mitglieder der Klassen Actinomyces, Streptococcus und Capnocytophaga wurden durch Metronidazol minimal beeinflußt. Jedoch verringerte Amoxicillin die Mengen und Verhältnisse von Actinomyces sp. während und nach der Therapie. Zusammenfassung: Die Daten suggerieren, daß Metronidazol und Amoxicillin in der schnellen Verringerung der Mengen von putativen parodontalen Pathogenen nützlich sind, daß dies aber durch andere Prozeduren begleitet wurden muß, um parodontale Stabilität zu erbringen. Résumé But: La présente recherche a évalué les modifications de niveaux et de proportions de 40 espèces bactériennes dans des prélèvements de plaque sous gingivale pendant, immédiatement après, et jusqu'à un an après un traitement par métronidazole ou amoxicilline associè avec le détartrage/surfaçage radiculaire. Méthode: Après avoir relevé les paramètres cliniques et microbiologiques initiaux, 17 sujets atteints de parodontite de l'adulte ont subi un détartrage/surfaçage radiculaire de toute la bouche et l'administration systémique pendant 14 jours de métronidazole (250 mg, 3× fois par jour, n=8) ou d'amoxicilline (500 mg, 3× par jour, n=9). Les mesures cliniques relevées initialement, à 90 jours, à 180 jours, et à 360 jours, étaient: le % de sites avec de la plaque, la rougeur gingivale, le saignement au sondage et la suppuration, la profondeur de poche (PD) et le niveau d'attache (AL). Des échantillons de plaque sous gingivale étaient prélevés sur la surface mésiale de toutes les dents, chez chaque sujet, initialement, à 90 jours, à 180 jours, et á 360 jours, et sur 2 dents postérieures choisies au hasard à 3, 7, et 14 jours pendant et après l'administration d'antibiotique. Le comptage de 40 expèces sous gingivales fut déterminé par la technique de l'hybridisation en damier DNA-DNA. La signification des différences au cours du temps fut déterminée par le test de Quade et entre les groupes par ANCOVA. Résultats: La profondeur moyenne des poches a étê réduite de 3.22±0.12 mm initialement à 2.81±0.16 mm (p<0.01) à 360 jours et de 3.38±0.28 mm à 2.80±0.14 mm (p<0.01) dans les groupes amoxicilline et metronidazole, respectivement. Les valeurs correspondantes pour AL étaient 3.21±0.30 à 2.76±0.32 (p<0.05) et 3.23±0.28 à 2.94±0.23 (p<0.01). Les niveau de B. forsythus, P. gingivalis et T. denticola, étaient fortement réduits pendant l'administration d'antibiotique et restaient plus bas à 360 jours qu'initialement. Les comptages (×105, ±SEM) de B. forsythus tombaient de niveaux initiaux de 0.66±0.16 à 0.04±0.02, 0.13±0.04, 0.10±0.03 et 0.42±0.19 dans le groupe amoxicilline à 14 jours, 90 jours, 180 jours, et 360 jours, respectivement (p<0.001). Les valeurs correspondantes pour les sujets traits par métronidazole étaient de: 1.69±0.28 à 0.02±0.01, 0.20±0.08, 0.22±0.06 et 0.22±0.08 (p<0.001). Les comptages des espèces Camopylobacter, Eubacterium nodatum, des espèces Fusobacterium nodatum, F. periodonticum et Prevotella nigrescensétaient également détectés à des niveaux moyens plus bas pendant, et immédiatement après traitement, mais augmentaient graduellement après cessation des antibiotiques. Les membres des genres Actinomyces, Streptococcus et Capnocytophagaétaient très peu affectés par le métronidazole. Par contre, l'amoxicilline diminuait les comptage et les proportions des Actinomyces pendant et après le traitement. Conclusions: Ces données suggèrent que le métronidazole et l'amoxicilline sont utiles pour diminuer rapidement les comptages des pathogènes parodontaux putatifs, mais qu'ils doivent être accompagnés d'autres procédés pour apporter une stabilité parodontale. [source]

    Relationship of cigarette smoking to the subgingival microbiota

    JOURNAL OF CLINICAL PERIODONTOLOGY, Issue 5 2001
    A. D. Haffajee
    Abstract Background: The relationship of cigarette smoking to the composition of the subgingival microbiota is not clear. Some studies indicated higher levels of certain species in smokers, while other studies failed to detect differences in the microbiota between subjects with different smoking histories. Thus, the purpose of the present investigation was to examine the prevalence, proportions and levels of the subgingival species in adult subjects who were current, past or never smokers. Method: 272 adult subjects ranging in age from 20,86 years with at least 20 teeth were recruited for study. Smoking history was obtained using a questionnaire. Clinical measures were taken at 6 sites per tooth at all teeth excluding third molars at a baseline visit. Subgingival plaque samples were taken from the mesial surface of all teeth excluding third molars in each subject at baseline and assayed individually for counts of 29 subgingival species using checkerboard DNA-DNA hybridization. Subjects were subset according to smoking history into never (n=124), past (n=98) and current smokers (n=50). Uni-variate and multi-variate analyses were used to seek associations between smoking category and the counts, proportions and prevalence of subgingival species. Results: Greater differences were observed for the prevalence (% of sites colonized) of the test species in the 3 smoking groups than were observed for counts or proportions of total counts. Members of the orange and red complexes including E. nodatum, F. nucleatum ss vincentii, P. intermedia, P. micros, P. nigrescens, B. forsythus, P. gingivalis and T. denticola were significantly more prevalent in current smokers than in the other 2 groups. The difference in prevalence between smokers and non-smokers was due to greater colonization at sites with pocket depth <4 mm. Stepwise multiple linear regression analysis indicated that combinations of the prevalence of 5 microbial species and pack years accounted for 44% of the variance for mean pocket depth (p<0.000001), while the prevalence of 3 microbial taxa along with age, pack years, current smoking and gender accounted for 31% of the variance in mean attachment level (p<0.000001). The difference in prevalence between current and never smokers of all members of the red complex and 8 of 12 members of the orange complex was significantly greater in the maxilla than in the mandible. Conclusions: The major difference between the subgingival microbiota in subjects with different smoking history was in the prevalence of species rather than counts or proportions. The greater extent of colonization in smokers appeared to be due to greater colonization at pocket depths <4 mm. Differences in colonization patterns between current and never smokers were greater in the maxilla than in the mandible. Zusammenfassung Grundlagen: Die Beziehung zwischen dem Zigarettenrauchen und der Zusammensetzung der subgingivalen Mikroflora ist nicht klar. Einige Studien verweisen auf höhere Titer von bestimmten Spezies bei Rauchern, während andere Studien keine Unterschiede in der Mikroflora zwischen Personen mit unterschiedlichem Raucher- oder Nichtraucherverhalten nachweisen konnten. Daher war der Zweck der vorliegenden Studie die Untersuchung von Prävalenz, Anteil und Titer der subgingivalen Spezies bei erwachsenen Patienten, die zur Zeit, früher oder niemals Raucher waren. Methode: Für die Studie wurden 272 erwachsene Patienten im Alter zwischen 20 und 86 Jahren und wenigstens 20 Zähnen rekrutiert. Die Anamnese des Rauchverhaltens wurde under Verwendung eines Fragebogens durchgeführt. Bei einer Eingangsuntersuchung erfolgten die klinischen Messungen an 6 Stellen pro Zahn bei allen Zähnen außer den dritten Molaren. Bei der Eingangsuntersuchung wurden, bei allen Zähnen außer den dritten Molaren, von den Mesialflächen subgingivale Plaqueproben entnommen. Für die einzelnen Flächen wurde die Anzahl von 29 subgingivalen Spezies mittels Schachbrett-DNA-DNA-Hybridisierung bestimmt. Die Patienten wurden entsprechend der Rauchervorgeschichte in folgende Gruppen eingeteilt: niemals (n=124), früher (n=98) und zur Zeit (n=50). Um Assoziationen zwischen den Rauchkategorien und der Anzahl, dem Anteil und der Prävalenz der subgingivalen Spezies herauszufinden wurden eine uni-variate und multi-variate Analyse verwendet. Ergebnisse: Es wurden größere Unterschiede zwischen den 3 Gruppen hinsichtlich der Prävalenz der Testspezies (% der Taschen die kolonisiert waren) beobachtet als bei der Anzahl oder dem Anteil an der Gesamtzahl der Keime beobachtet wurde. Die Prävalenz der Keime des orangen und roten Komplexes einschließlich. E. nodatum, F. nucleatum ss vincentii, P. intermedia, P. micros, P. nigrescens, B. forsythus, P. gingivalis und T. denticola war bei den aktuellen Rauchern stärker prävalent als in den anderen beiden Gruppen. Die Differenz in der Prävalenz zwischen Rauchern und Nichrauchern wurde verursacht durch eine stärkere Kolonisation in Taschen mit einer Taschentiefe <4 mm. Die schrittweise multiple lineare Regressionsanalyse zeigte, dass Kombinationen der Prävalenz von 5 mikrobiellen Spezies und der Packungsjahre für 44% der Varianz der mittleren Taschentiefe verantwortlich waren (p<0.000001), während die Prävalenz von 3 mikrobiellen Taxa zusammen mit Alter, Packungsjahre, Raucherstatus und Geschlecht für 31% der Varizna im mittleren Attachmentniveau verantwortlich waren (p<0.000001). Die Differenz in der Prävalenz zwischen den aktuellen Rauchern und den die niemals rauchten war für alle Keime der roten Komplexes und 8 von 12 Keimen des orangen Komplexes im Oberkiefer signifikant größer als im Unterkiefer. Schlussfolgerung: Der Hauptunterschied zwischen der subgingivalen Mikroflora bei Patienten mit unterschiedlicher Rauchervorgeschichte lag mehr bei der Prävalenz der Spezies als bei der Anzahl der Keime oder den Anteilen an der Gesamtflora. Das größere Maß an Kolonisation bei den Rauchern schien durch eine stärkere Kolonisation in Taschen <4 mm verursacht zu sein. Differenzen im Kolonisationsmuster zwischen aktuellen Rauchern und Nichtrauchern die niemals rauchten waren im Oberkiefer größer als im Unterkiefer. Résumé Origine, but: La relation entre l'usage de la cigarette et la composition de la microflore sous gingivale n'est pas claire. Certaines études indiquent d'importants niveaux de certaines espèces chez les fumeurs, alors que d'autres études n'arrivent pas à détecter de différences dans la micrflore entre des sujets ayant des histoires tabagiques différentes. Aussi, le propos de cette recherche est d'examiner la prévalence, les proportions et le niveau des espèces sous gingivales chez des sujets adultes fumeurs, anciens fumeurs ou non-fumeurs. Méthodes: 272 sujets adultes, âgès de 20 à 86 ans, ayant au moins 20 dents furent recrutés pour l'étude. L'histoire tabagique fut obtenue à l'aide d'un questionnaire. Des mesures cliniques furent prises sur 6 sites par dents, sur toutes les dents à l'exception des troisièmes molaires lors de la première visite. Des échantillons de plaque sous gingivale étaient prélevés sur la face mésiale de chaque dent à l'exception des troisièmes molaires chez chaque sujet lors de la première visite et individuellement testés pour le comptage de 29 espèces sousgingivales par hybridisation en damier ADN-ADN. Les sujets étaient groupés en sous ensembles en fonction de leur histoire tabagique en non-fumeurs (n=124), ancien fumeurs (n=98), et fumeurs (n=50). Des analyses monovariées et multivariées furent utilisées pour rechercher des associations entre les catégories de fumerus et les comptages, proportions et prévalences des espèces bactériennes. Résultats: De plus grandes différences étaient observées pour la prévalence (% de sites colonisés) des expèces testées dans les 3 groupes, que pour le comptage ou la proportion des comptages totaux. Les membres des complexes orange et rouge dont E. nodatum, F. nucléatum ss vicentii, P. intermedia, P. micros, P. nigrescens, B. forsythus, P. gingivalis, et T. denticolaétait significativement plus prévalent chez les fumeurs que dans les 2 autres groupes. La différence de prévalence entre les fumeurs et les non-fumeurs était due à une plus grande colonisation des sites dont la profondeur de poche était <4 mm. L'analyse par régression linéaire multiple stepwise indiquait que les combinaisons de la prévalence de 5 espèces microbiennes et les paquets-années comptaient pour 44% de la variance pour la moyenne de profondeur de poche (p<0.000001), alors que la prévalence de 3 taxons microbiens avec l'âge, les paquets-années, le tabagisme présent et le sexe comptaient pour 31% de la variance pour le niveau d'attache moyen (p<0.000001). La différence de prévalence entre les fumerus en activité et les non-fumeurs (jamais fumé) de tous les membres du complexe rouge et de 8 des 12 membres du complexe orange était significativement plus élevée au maxillaire qu'à la mandibule. Conclusions: La différence majeure entre les microflores sous gingivales chez les sujets ayant des histoires tabagiques différentes se trouvaient dans la prévalence des expèces plutôt que dans leurs quantité ou leurs proportions. La plus grande importance de colonisation chez les fumerus apparaît être dûe à une colonisation plus grande dans les poches <4 mm. Des différences des caractéristiques de colonisation entre les fumerus actifs et les personnes n'ayant jamais fuméétaient plus importantes au maxillaire qu'à la mandibule. [source]

    Microbial composition of supra- and subgingival plaque in subjects with adult periodontitis

    JOURNAL OF CLINICAL PERIODONTOLOGY, Issue 10 2000
    Laurie Ann Ximénez-Fyvie
    Abstract Background, aims: The purpose of the present study was to compare and relate the microbial composition of supra and subgingival plaque in 23 adult periodontitis subjects (mean age 51±14 years). Methods: A total of 1,170 samples of supra and subgingival plaque were collected from the mesial aspect of every tooth (up to 28 supra and 28 subgingival samples) from each subject and evaluated for the presence and levels of 40 bacterial taxa using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical assessments including dichotomous measures of gingival redness, bleeding on probing, plaque accumulation and suppuration, as well as duplicate measures of pocket depth and attachment level, were made at 6 sites per tooth. The counts (levels), % DNA probe count (proportion) and % of sites colonized (prevalence) of each species in supra and separately in subgingival plaque were computed for each subject. Significance of differences between supra and subgingival plaque for each species was sought using the Wilcoxon signed ranks test and adjusted for multiple comparisons. Results: All 40 taxa were detected in both supra and subgingival plaque. Actinomyces species were the most prevalent taxa in both habitats. 75 to 100% of supra and 62 to 100% of subgingival sites were colonized by at least one of the 5 Actinomyces species. Supragingival samples exhibited significantly higher counts of Actinomyces naeslundii genospecies 1, Actinomyces israelii, Actinomyces odontolyticus, Neisseria mucosa, Streptococcus gordonii, Capnocytophaga ochracea and Capnocytophaga sputigena when compared with mean counts in subgingival samples taken from the same tooth surfaces. Subgingival plaque samples presented significantly higher counts of Prevotella nigrescens, Prevotella intermedia, Bacteroides forsythus and Porphyromonas gingivalis. Subgingival samples exhibited a significantly higher proportion of "red" and "orange complex" species, while supragingival plaque exhibited higher proportions of "green" and "purple" complex species as well as Actinomyces species. Suspected periodontal pathogens could be detected in supragingival plaque from sites where subgingival samples were negative for the same species. Conclusions: The data indicate that supragingival plaque can harbor putative periodontal pathogens, suggesting a possible rôle of this environment as a reservoir of such species for the spread or reinfection of subgingival sites. [source]

    The effect of repeated professional supragingival plaque removal on the composition of the supra- and subgingival microbiota

    JOURNAL OF CLINICAL PERIODONTOLOGY, Issue 9 2000
    Laurie Ann Ximénez-Fyvie
    Abstract Background, aims: The purpose of the present investigation was to determine the effect of weekly professionally administered supragingival plaque removal on the composition of the supra and subgingival microbiota. Methods: 18 adult subjects with periodontitis who had been treated and were in a maintenance phase of therapy were clinically and microbiologically monitored at baseline, 3, 6 and 12 months. After the baseline visit, the subjects received scaling and root planing followed by professional supragingival plaque removal every week for 3 months. Clinical measures of plaque accumulation, bleeding on probing (BOP), gingival redness, suppuration, pocket depth and attachment level were made at 6 sites per tooth at each visit. Separate supra (N=1804) and subgingival (N=1804) plaque samples were taken from the mesial aspect of all teeth excluding third molars in each subject at each time point and evaluated for their content of 40 bacterial taxa using checkerboard DNA-DNA hybridization. Significance of changes in mean counts, prevalence and proportions of bacterial species over time in both supra and subgingival samples were determined using the Quade test and adjusted for multiple comparisons. Results: Mean % of sites exhibiting plaque, gingival redness and BOP were significantly reduced during the course of the study. Significant decreases in mean counts were observed in both supra and subgingival samples. Mean total DNA probe counts (×105, ±SEM) at baseline, 3, 6 and 12 months were: 133±19, 95±25, 66±6, 41±6 (p<0.001) for supragingival samples and 105±22, 40±10, 19±4, 13±3 (p<0.001) for subgingival samples. Mean counts of 22 of 40 and 34 of 40 species tested were significantly reduced in the supra and subgingival samples respectively over the monitoring period. For example, mean counts of Porphyromonas gingivalis×105 at baseline, 3, 6 and 12 months in the subgingival plaque samples were 2.0±0.4, 0.5±0.2, 0.6±0.3, 0.3±0.1 (p<0.001); Bacteroides forsythus 2.0±0.6, 0.4±0.1, 0.4±0.2, 0.1±0.2 (p<0.001); Treponema denticola 3.4±1.1, 0.8±0.3, 0.4±0.2, 0.3±0.3 (p<0.01). Similar reductions were seen in supragingival plaque samples. While counts were markedly reduced by professional plaque removal, the proportion and prevalence of the 40 test species were marginally affected. Conclusions: Weekly professional supragingival plaque removal profoundly diminished counts of both supra- and subgingival species creating a microbial profile comparable to that observed in periodontal health. This profile was maintained at the final monitoring visit, 9 months after completion of therapy. [source]

    The lower genital tract microbiota in relation to cytokine-, SLPI- and endotoxin levels: application of checkerboard DNA-DNA hybridization (CDH),

    APMIS, Issue 4 2008
    NATALIA NIKOLAITCHOUK
    In the present study the lower genital tract microbiota in asymptomatic fertile women (n=34) was identified and quantified by culturing vaginal secretions. Also, vaginal and cervical samples were analyzed by a semiquantitative checkerboard DNA-DNA hybridization technique (CDH) based on genomic probes prepared from 13 bacterial species (Bacteroides ureolyticus, Escherichia coli, Fusobacterium nucleatum, Gardnerella vaginalis, Mobiluncus curtisii ss curtisii, Prevotella bivia, Prevotella disiens, Prevotella melaninogenica, Atopobium vaginae, Lactobacillus iners, Staphylococcus aureus ss aureus, Streptococcus anginosus, and Streptococcus agalactiae). The bacterial species found by either culture or CDH were correlated with proinflammatory cytokines (IL-1,, IL-1,, IL-6, IL-8), secretory leukocyte protease inhibitor (SLPI), and endotoxin in the cervicovaginal samples. Grading the women into healthy, intermediate, or bacterial vaginosis (BV) as based on Gram staining of vaginal smears, the viable counts of lactobacilli (L. gasseri) and of streptococci-staphylococci combined were highest in the intermediate group. In BV, particularly the high concentrations of Actinomyces urogenitalis, Atopobium vaginae, and Peptoniphilus harei were noted (,1011 per ml). The total viable counts correlated with both cervical IL-1, and IL-1,. A strong negative correlation was observed between L. iners and total viable counts, G. vaginalis, or cervical IL-1,, while it correlated positively with SLPI. Analysis of vaginal and cervical samples from 26 out of the 34 women by CDH showed that anaerobic bacteria were more frequently detected by CDH compared to culture. By this method, A. vaginae correlated with G. vaginalis, and L. iners with S. aureus. With regard to cytokines, B. ureolyticus correlated with both cervical and vaginal IL-1, as well as with cervical IL-8, while F. nucleatum, S. agalactiae, S. anginosus, or S. aureus correlated with vaginal IL-1,. Furthermore, all Gram-negative bacteria taken together, as measured by CDH, correlated with vaginal endotoxin and inversely with vaginal SLPI. The significance of the results is discussed. In summary, mapping of the identity and quantity of vaginal bacterial species and their association with locally produced host innate immune factors will help in defining various types of abnormal vaginal microbiota, developing new ways of assessing the risk of ascending subclinical infections, and in treating them. CDH appears to be a suitable tool for future analyses of large numbers of clinical samples with an extended number of bacterial probes. [source]

    Phylogeny of major lineages of suboscines (Passeriformes) analysed by nuclear DNA sequence data

    JOURNAL OF AVIAN BIOLOGY, Issue 1 2001
    Martin Irestedt
    Phylogenetic relationships among major groups of passeriform birds were studied by analyses of nucleotide sequence data from two nuclear genes, c- myc and RAG-1. The results corroborated both the monophyly of the order Passeriformes, and the major dichotomy into oscine and suboscine passerines previously suggested based on syringeal morphology and DNA-DNA hybridizations. The representatives of the Old World suboscines (families Eurylaimidae, Philepittidae and Pittidae) formed a monophyletic clade. The New World suboscines clustered into two clades. The first contained Conopophaga (Conopophagidae), Furnarius (Furnariidae), Lepidocolaptes (Dendrocolaptidae), Thamnophilus (Formicariidae), and Rhinocrypta (Rhinocryptidae). Previously, the monophyly of this group has been inferred from their possession of a unique, "tracheophone" syrinx, and from DNA-DNA hybridisation data. The second clade of New World suboscines includes Gubernetes and Muscivora (Tyrannidae), Phytotoma (Phytotomidae), Tityra (Cotingidae) and Pipra (Pipridae). This group of families have been considered monophyletic based on morphology (although ambiguously) and DNA-DNA hybridisation. The sister group relationship of Tityra and Phytotoma supports the previously supposed cotingid affinity of Phytotoma. Nuclear DNA data also unambiguously group the lyrebirds Menura with the oscines. The presented results from the analysis of nuclear DNA agree well with morphology and DNA-DNA hybridisation data. The precise age of the divergences studied herein are unknown but based on interpretations of the fossil record of passerine birds many of them might date back to the early Tertiary. The agreement between data from the nuclear DNA and other sources, along with the fact that neither of the studied genes showed sign of saturation, indicate the great potential of these two nuclear genes to resolve very old divergences in birds. [source]