Effects of the Irrigation Needle Design on Root Canal Disinfection and Cleaning.

INTRODUCTION: This ex vivo study evaluated the disinfecting and cleaning effects of root canal preparation using sodium hypochlorite irrigation with 3 different needle designs. METHODS: Mesial roots from extracted mandibular molars with Vertucci class II configuration were anatomically matched based on micro-computed tomography (micro-CT) analyses and distributed into 3 groups (n = 18/group). The canals were contaminated with a mixed bacterial culture for 30 days and then subjected to preparation using 2.5% sodium hypochlorite irrigation with open-ended flat needle (3 mm short of the working length - WL), closed-ended side-vented irrigation needle (1 mm short of the WL), or a closed-ended plastic needle with 2 lateral outlets (TruNatomy) (1 mm short of the WL). Bacteriological samples were taken from the canals before (S1) and after preparation (S2). After another micro-CT scan, the roots were sectioned and samples were also taken from the apical canal segment (S3). Bacterial reduction was assessed by quantitative real-time polymerase chain reaction. The amount of accumulated hard tissue debris was evaluated by micro-CT. RESULTS: A substantial reduction in bacterial counts was observed in all 3 groups when comparing S1 with S2 (99.95%, 98.93%, and 98.90% in the open-ended, closed-ended, and TruNatomy needle groups, respectively) (P < .01). No significant differences were observed between groups for bacterial reduction in the full and apical canal (P > .05). There were no intergroup differences in the amount of accumulated hard tissue debris either (P > .05). The open-ended needle group showed significantly more specimens with quantitative real-time polymerase chain reaction negative results for bacteria in S3 than TruNatomy (P < .05). CONCLUSION: When used up to an appropriate insertion depth, the 3 needle types result in similar antibacterial and cleaning performance, provided variables such as needle size and irrigant type, volume and flow rate are controlled. Exclusive analysis of the apical segment, including the isthmus, revealed that the open-ended needle yielded more cases negative for bacteria.

Disinfection and Shaping of Vertucci Class II Root Canals after Preparation with Two Instrument Systems and Supplementary Ultrasonic Activation of Sodium Hypochlorite.

INTRODUCTION: This study compared disinfection and shaping after root canal preparation with either XP-endo Shaper or TruNatomy instrument systems, supplemented by ultrasonic activation of sodium hypochlorite (NaOCl) with either stainless-steel (SS) or nickel-titanium (NiTi) inserts. METHODS: Mesial roots from mandibular molars with Vertucci class II configuration were divided into 2 groups (n = 24) based on anatomically paired micro-computed tomography (micro-CT) analyses. Pre and postpreparation micro-CT scans were obtained to evaluate the shaping performance. The canals were contaminated with a mixed bacterial culture for 30 days and then subjected to preparation with either XP-endo Shaper or TruNatomy instruments using NaOCl irrigation. Supplementary ultrasonic activation of NaOCl was conducted using either an SS (TruNatomy group) or NiTi (XP-endo Shaper group) insert. Bacteriological samples were taken from the canals before preparation (S1), after preparation (S2), and after the supplementary approach (S3). Bacterial reduction was evaluated using a quantitative real-time polymerase chain reaction. RESULTS: Preparation with both instrument systems significantly reduced bacterial counts (P < .01). After preparation, 36% (TruNatomy) and 35% (XP-endo Shaper) were negative for bacteria. These values increased to 59% and 65% after ultrasonic activation with the SS and NiTi inserts, respectively. The quantitative data in S2 showed that XP-endo Shaper promoted a significantly higher bacterial reduction than TruNatomy (P < .05). No significant intragroup differences were observed after ultrasonic activation (P > .05), probably because the SS insert promoted a significantly higher S2-to-S3 reduction than the NiTi insert (P < .01). Micro-CT analysis revealed no significant differences in the unprepared areas between the groups (P > .05). CONCLUSIONS: The XP-endo Shaper caused a significantly higher bacterial reduction than TruNatomy in Vertucci class II canals. Better antibacterial results after ultrasonic activation were observed for the SS ultrasonic inserts than for the NiTi inserts.

Enhancing the Intracanal Antibacterial Effects of Sodium Hypochlorite with Etidronic Acid or Citric Acid.

INTRODUCTION: This study evaluated the bacterial reduction promoted by root canal preparation using irrigation with sodium hypochlorite (NaOCl) alone, associated with etidronic acid (1-hydroxyethane 1,1-diphosphonic acid [HEDP]) or alternated with citric acid, and after a supplementary agitation step. METHODS: Extracted mandibular premolars were selected and distributed into 3 groups based on anatomically paired micro-computed tomographic analyses. The canals were contaminated with Enterococcus faecalis for 30 days and then subjected to chemomechanical preparation with a reciprocating instrument under irrigation with NaOCl alone, mixed with HEDP (NaOCl/HEDP), or alternated with citric acid (NaOCl/CA). A supplementary agitation step with the XP-endo Finisher was performed in all groups. Intracanal bacteriological samples were taken before (S1) and after preparation (S2) and after the supplementary approach (S3). DNA was extracted from the samples and subjected to quantitative real-time polymerase chain reaction. RESULTS: Intragroup analyses revealed a substantial bacterial reduction from S1 to S2 or S3 in all groups (P < .01). The supplementary agitation resulted in S2-to-S3 bacterial reduction of 6%, 68%, and 80% in the NaOCl, NaOCl/HEDP, and NaOCl/CA groups, respectively. Irrigation with NaOCl alone resulted in 53% and 47% of samples negative for bacteria in S2 and S3, respectively. Corresponding figures for NaOCl/HEDP were 75% and 85%, and 44% and 72% for NaOCl/CA. Intergroup analyses of S2 samples showed that NaOCl/HEDP was significantly more effective than the other 2 in reducing the bacterial levels (P < .05). After the supplementary approach, both NaOCl/HEDP and NaOCl/CA were significantly more effective than NaOCl alone (P < .05), with no significant differences between them (P > .05). CONCLUSIONS: Both the freshly combined NaOCl/HEDP solution and the alternate use of NaOCl and citric acid followed by XP-endo Finisher agitation resulted in significantly higher intracanal bacterial reduction than NaOCl alone.

Disinfecting and Shaping Type I C-shaped Root Canals: A Correlative Micro-computed Tomographic and Molecular Microbiology Study.

INTRODUCTION: This ex vivo study compared the disinfection and shaping abilities of 2 preparation protocols in C-shaped canals. METHODS: Mandibular second molars with type I C-shaped canals were pair matched based on micro-computed tomographic (micro-CT) analysis and distributed into 2 groups. The canals were contaminated with a mixed bacterial culture and prepared using 2 protocols, both with 2.5% sodium hypochlorite irrigation: XP-endo Shaper (FKG Dentaire, La Chaux-de-Fonds, Switzerland) supplemented with XP-endo Finisher (FKG Dentaire) (XP-E) and BioRaCe (FKG Dentaire) supplemented with a Hedström file (Dentsply/Sirona, Tulsa, OK) (BR-Hed). Micro-CT scans and intracanal bacteriologic samples were taken before (scan #1 and sample 1) and after preparation (scan #2 and sample 2) and after the supplementary step (scan #3 and sample 3). Canal shaping was evaluated by micro-CT imaging, and disinfection was assessed by quantitative polymerase chain reaction. RESULTS: Micro-CT data from the XP-E (n = 21) and BR-Hed (n = 23) protocols revealed no significant differences between groups regarding shaping parameters (canal volume, surface area, structure model index, and prepared walls) after preparation and after the supplementary step (P > .05). All these parameters significantly increased after each preparation step (P < .05), except for structure model index changes after the XP-endo Finisher (P > .05). Bacteriologic data from the XP-E (n = 21) and BR-Hed (n = 22) groups showed that 14 (66.7%) and 10 (45.5%) S2 samples still had detectable bacteria, respectively. The corresponding figures for supplementary steps with the XP-endo Finisher and Hedström file were 11 (52.4%) and 10 (45.5%), respectively. Bacterial counts in sample 1 significantly decreased in samples 2 and 3 in both groups (P < .01). Intragroup comparison between sample 2 and sample 3 showed a 66% reduction in counts after using the Hedström file and 18% after using the XP-endo Finisher (P > .05 for both groups). The quantitative bacterial reduction was not different between groups (P > .05). CONCLUSIONS: The XP-endo Shaper and BioRaCe systems have similar disinfecting and shaping abilities in C-shaped canals. Supplementary steps with the Hedström file and the XP-endo Finisher were similarly effective in improving shaping, but this was not sufficient to enhance disinfection. About half of the cases, in both groups, still harbored detectable amounts of bacteria.

Impact of Contracted Endodontic Cavities on Root Canal Disinfection and Shaping.

INTRODUCTION: The impact of minimally invasive endodontic procedures on root canal disinfection has not been determined. This ex vivo study compared root canal disinfection and shaping in teeth with contracted or conventional endodontic cavities. METHODS: Mandibular incisors with oval-shaped canals were selected and anatomically matched based on micro-computed tomographic (micro-CT) analysis and distributed into 2 groups. Conservative and conventional access cavities were prepared, and the canals were contaminated with a pure culture of Enterococcus faecalis for 30 days. Root canal preparation in both groups was performed using the XP-endo Shaper instrument (FKG Dentaire, La Chaux-de-Fonds, Switzerland) and 2.5% sodium hypochlorite irrigation. Intracanal bacteriologic samples were taken before and after preparation, and DNA was extracted and subjected to quantitative polymerase chain reaction. Micro-CT scans taken before and after preparation were used for shaping evaluation. Bacteriologic data were analyzed by the Poisson regression model and the chi-square test with Yates correction. Micro-CT data were analyzed by the Wilcoxon, Mann-Whitney, and Student t tests with the significance level set at 5%. RESULTS: All initial samples were positive for E. faecalis. After preparation, the number of bacteria-positive samples was significantly higher in the contracted cavity group (25/29, 86%) than in the conventional cavity group (14/28, 50%) (P < .01). Intergroup quantitative comparison showed that the reduction in bacterial counts was also significantly higher in the group of conventional cavities (P < .01). Micro-CT data revealed no significant difference in the amount of unprepared areas between groups. CONCLUSIONS: Our findings showed that although shaping using an adjustable instrument was similar between groups, disinfection was significantly compromised after root canal preparation of teeth with contracted endodontic cavities.

Root Canal Disinfection by Single- and Multiple-instrument Systems: Effects of Sodium Hypochlorite Volume, Concentration, and Retention Time.

INTRODUCTION: This ex vivo study evaluated the intracanal bacterial reduction promoted by chemomechanical preparation using a single-file technique varying the volume, concentration, and retention time of sodium hypochlorite (NaOCl) irrigation in comparison with a multifile system. METHODS: Palatal roots from extracted maxillary first molars were selected and anatomically matched based on microcomputed tomographic analysis for group distribution. The canals were contaminated with a fresh mixed bacterial culture grown in anaerobiosis and recently obtained from a tooth with apical periodontitis. Specimens were divided into 4 groups of 24 each according to the following preparation protocols: REC-6LOW (Reciproc R50 instrument [VDW, Munich, Germany], 6% NaOCl, low irrigant volume), REC-2.5LOW (R50, 2.5% NaOCl, low irrigant volume), REC-2.5HI (R50, 2.5% NaOCl, high irrigant volume), and BR-2.5HI (BioRaCe [FKG Dentaire, LaChaux-de-Fonds, Switzerland], 2.5% NaOCl, high irrigant volume). The total time of preparation was recorded. Intracanal bacteriologic samples were taken before and after preparation; DNA was extracted and subjected to quantitative polymerase chain reaction. RESULTS: Bacteria were detected in 22 initial samples from the REC-2.5LOW group and in 23 from the other groups. Intragroup analysis showed that all tested preparation protocols were highly effective in significantly reducing the intracanal bacterial counts (P < .001). Intergroup comparison of bacterial reduction levels revealed a statistically significant difference between BR-2.5HI and REC-2.5LOW (P < .05). Counts of bacteria were 2.5 times significantly higher in REC-2.5LOW compared with BR-2.5HI. No other significant differences were found in quantitative findings (P > .05). CONCLUSIONS: The concerted effects of multiple instruments, the high volume of irrigation, and the long retention time of NaOCl irrigant had a positive influence on intracanal disinfection during chemomechanical preparation.

Passive ultrasonic irrigation in root canal: systematic review and meta-analysis.

OBJECTIVE: To investigate whether there are differences between the root canal disinfection, comparing the passive ultrasonic irrigation technique with the conventional technique. MATERIALS AND METHODS: The following electronic databases were searched: Pubmed; VHL; Web of Sciences and OVID with no publication date restriction. The study's quality evaluation was carried out using the Handbook by Cochrane. The online research identified 5464 studies. From the nine studies selected for a full reading of the text, five were included in the present systematic review. Meta-analysis was performed in three articles, which evaluated the root canal's cleanness through microbiological analysis. RESULTS: Only one article concluded that the ultrasonic passive irrigation showed a better performance compared with the conventional irrigation. None of the articles analyzed presented a low risk of bias in all domains. According to the results of the meta-analysis, there was no statistical difference between the groups (OR = 0.34, IC 95%: 0.10-1.19). CONCLUSIONS: The level of evidence comparing the two techniques is fragile since in all studies some type of bias was observed which may interfere in the results and conclusions.

Does adding an instrument after root preparation with Reciproc® R25 increase bacterial reduction?

AIM: The aim of this study was to compare the bacterial reduction achieved with reciprocating instruments such as Reciproc® R25 (VDW GMBH, Munich, Germany) and Reciproc® R25 combined with rotary file Mtwo 40.04 (VDW GMBH) during root canal preparation. MATERIALS AND METHODS: Fifty mesiobuccal root canals of maxillary molars were contaminated with Enterococcus faecalis broth culture and then an initial bacterial sample was collected from the root canal with paper cones and plated on brain-heart infusion agar. The root canals were divided into four groups as follows: Reciproc® R25 (n = 20) and Reciproc® R25 combined with Mtwo 40.04 instrument (n = 20). The negative controls consisted of five uncontaminated root canals and the positive control consisted of five contaminated roots that were not subjected to any decontamination procedure. Irrigation was performed using sodium chloride. After instrumentation, samples were collected with paper cones and the rate of bacterial reduction was calculated. Microbiological testing (colony-forming units [CFUs]) was performed to quantify the decontamination obtained by the proposed protocols. Statistical analysis was performed by paired t-test and analysis of variance test. RESULTS: Both techniques significantly reduced the number of bacteria in the root canal (P < 0.05), with no significant difference among them (P > 0.05). CONCLUSION: The addition of the instrument Mtwo 40.04 after the root preparation with the instrument R25 does not improve its decontamination capability.

Clinical Antibacterial Effectiveness of Root Canal Preparation with Reciprocating Single-instrument or Continuously Rotating Multi-instrument Systems.

INTRODUCTION: This in vivo study compared the antibacterial effectiveness of a reciprocating single-instrument system (Reciproc; VDW, Munich, Germany) and a rotary multi-instrument system (BioRaCe; FKG Dentaire, La Chaux-de-Fonds, Switzerland) during the preparation of infected root canals of teeth with primary apical periodontitis. METHODS: Root canals from single-rooted teeth with necrotic pulps and apical periodontitis were instrumented using either Reciproc (n = 29) or BioRaCe (n = 30) instruments under irrigation with 2.5% sodium hypochlorite. DNA was extracted from samples taken before and after preparation and subjected to quantitative analysis of total bacteria and streptococci by using the quantitative real-time polymerase chain reaction. RESULTS: All initial samples were positive for the presence of bacteria, with median numbers of 7.1 × 10(5) and 1.31 × 10(5) bacterial cells for the Reciproc and BioRaCe groups, respectively. After preparation with Reciproc and BioRaCe, 16 (55%) and 15 (50%) root canals still had detectable bacteria with median counts of 7.05 × 10(2) and 6.03 × 10(1), respectively. Both systems were highly effective in reducing the total bacterial counts (P < .001), and there were no significant differences between them (P > .05). Streptococci were highly frequent, and both systems succeeded in significantly reducing their levels (P < .001). CONCLUSIONS: Both reciprocating single-instrument and rotary multi-instrument systems were highly effective in reducing the counts of total bacteria and streptococci in root canals of teeth with apical periodontitis. Regardless of the system used, approximately one half of the teeth still had detectable bacteria.

Endotoxinas nas infecções endodônticas: revisão sistemática com metanálise e ensaio clínico randomizado / Endotoxins in endodontic infection: a systematic review with metaanalysis and a clinical trial

Sabe-se que as endotoxinas provenientes das bactérias grã-negativas desempenham um importante papel nas infecções endodônticas se relacionando à sinais e sintomas clínicos/radiográficos. Sendo assim, a redução ou eliminação de endotoxinas é fundamental para a resolução da inflamação periapical. Os objetivos do presente estudo foram: Artigo 1 - realizar uma revisão sistemática com metanálise com o intuito de avaliar a relação entre níveis de endotoxinas e a presença de sinais e sintomas clínicos, bem como de sinais radiográficos em pacientes com infecção endodôntica primária; Artigo 2 - fazer um ensaio clínico randomizado para avaliarmos estratégias clínicas para otimizar a eliminação microbianana e de lipopolissacarídeos (LPS) utilizando a Terapia Fotodinâmica (PDT) como tratamento suplementar ao PQM nas modalidades de terapia endodôntica em única e múltiplas sessões. Métodos: Artigo 1 - para a revisão sistemática uma busca eletrônica foi realizada por dois autores, no idioma inglês, nas bases de dados Medline/PubMed, Embase, Cochrane Library, Scielo, Science Direct, Web of Knowledge e Scopus. Após seleção dos artigos foi realizado uma metanálise com análises de sinais e sintomas clínicos/radiográficos. Artigo 2 - Selecionou-se 24 casos de dentes com infecção endodôntica primária que foram aleatoriamente divididos em 2 grupos (n=12): SU - sessão única e SM ­ sessão múltipla. Foram realizadas coletas microbiológica e de endotoxinas dos canais radiculares utilizando cone de papel estéril/ apirogênico em diferentes momentos operatórios. Foi realizado cultura microbiana para determinar a carga bacteriana em UFC/mL (Unidades Formadoras de Colônias/mL) e para quantificação de endotoxinas foi realizado o teste KQCL de "Lisado de Amebócito Limulus" (LAL). Resultados: Artigo 1 - A metanálise revelou que individuos com dor à percussão (TTP) (P=0,04; I2 57%) e episódio de dor prévio (PEP) (P=0,001; I2 81%) apresentaram maior nível de endotoxina que os casos que não apresentaram nenhum destes sintomas. O tamanho da área radiolucente (SRL) > 2 mm (P=0,02; I2 68%) e presença de exudação (EX) (P=0,0007; I2 0%) estavam associados com maior nível de endotoxinas. Artigo 2 - Para carga microbiana, a suplementação do tratamento em sessão única com PDT reduziu significativamente os níveis bacterianos (p<0,05), mas não nos casos de duas sessões (p>0,05). Para os níveis de endotoxinas, na sessão única e de duas sessões, não houve diferença significativa após uso de PDT (p>0,05). Foi possível concluir que Artigo 1 ­ A metanálise mostrou forte evidência que endotoxina está relacionada com a presence de sinais e sintomas clínicos/radiográficos em pacientes com infecção endodôntica. Artigo 2 - A terapia fotodinâmica melhorou o desempenho da desinfecção bacteriana dos canais radiculares em uma visita, mas não para a modadilidade de 2 visitas após uso de medicação à base de Ca(OH)2. PDT não foi efetivo contra endotoxinas(AU)

Endotoxins originating from gram negatives bacterias are related to signals and clinical/radiographic symptoms. Therefore, reduction or elimination of endotoxins is essential to solve periapical inflammation. The aim of this dissertation was: Article 1 - To relate endotoxin levels and presence of clinical signs/symptoms and radiographic features in endodontic infection. Article 2 - To evaluate the effectiveness of supplemental photodynamic therapy (PDT) in optimizing the removal of bacteria and endotoxins from primarily infected root canals after one-visit and twovisit treatments. Methodology: Article 1 - Electronic searches were performed on Medline/PubMed, Embase, Cochrane Library, Scielo, Science Direct, Web of Knowledge and Scopus databases for identification of relevant studies published up to July 2016. Only reports in English were included. The selected literature was reviewed by two authors and classified as either suitable or unsuitable for inclusion in this review. The relationship between endotoxin levels and presence of clinical signs/symptoms and radiographic features were determined. Additionally, a metaanalysis was performed. Article 2 - 24 primarily infected root canals with apical periodontitis were selected and randomly divided into one-visit (n=12) and two-visit (n=12). Samples were collected before and after root canal procedures. Endotoxins were quantified by chromogenic limulus amebocyte lysate assay. Culture techniques were used to determine bacterial colony-forming unit counts. The systematic review and meta-analysis aimed to evaluate the relationship between endotoxin levels and presence of clinical signs/symptoms and radiographic features in patients with endodontic infection. Results: Article 1 - Among the 285 articles identified in the initial search, 29 were included for full-text appraisal and only eight studies met the inclusion criteria for this systematic review. Meta-analysis revealed that individuals having teeth with tenderness to percussion (TTP) (P=0.04; I2 57%) and previous episode of pain (PEP) (P=0.001; I2 81%) had higher levels of endotoxin than their counterparts. Size of radiographic lesion (SRL) > 2 mm (P=0.02; I2 68%) and presence of root canal exudation (EX) (P=0.0007; I2 0%) were associated with higher levels of endotoxin. Article 2 - Bacteria and endotoxins were detected in 100% of the initial samples, with median values of 1.97 x 105 CFU/mL and 24.983 EU/mL, respectively. The CMP using single-file reciprocating technique was effective in the reduction of bacteria and endotoxins (All, p < .05). The supplemental PDT was effective in reducing bacterial load in the one-visit (p < .05) but not in the two-visit treatment after use of Ca(OH)2 medication for 14 days (p>.05). In the two-visit group, after 14 days of inter- appointment medication with Ca(OH)2, a significant reduction in the median levels of endotoxins was found in comparison to CMP alone (from 1.041 to 0.094 EU/mL)(p < .05). Despite the type of treatment, the supplemental PDT was not effective against endotoxins (p>.05). Conclusion: Article 1 - This meta-analysis provides strong evidence that endotoxin are related with the presence of clinical signs/symptoms and radiographic features in patients with endodontic infection. Article 2 - The photodynamic therapy optimized the disinfection of bacteria from root canals in one-visit but not for two visit treatment modality with the accomplishment of calcium hydroxide medication. Thus, PDT is not effective against endotoxins(AU)

Does the Reciproc file remove root canal bacteria and endotoxins as effectively as multifile rotary systems?

AIM: To evaluate the effectiveness of Reciproc for the removal of cultivable bacteria and endotoxins from root canals in comparison with multifile rotary systems. METHODOLOGY: The root canals of forty human single-rooted mandibular pre-molars were contaminated with an Escherichia coli suspension for 21 days and randomly assigned to four groups according to the instrumentation system: GI - Reciproc (VDW); GII - Mtwo (VDW); GIII - ProTaper Universal (Dentsply Maillefer); and GIV -FKG Race(™) (FKG Dentaire) (n = 10 per group). Bacterial and endotoxin samples were taken with a sterile/apyrogenic paper point before (s1) and after instrumentation (s2). Culture techniques determined the colony-forming units (CFU) and the Limulus Amebocyte Lysate assay was used for endotoxin quantification. Results were submitted to paired t-test and anova. RESULTS: At s1, bacteria and endotoxins were recovered in 100% of the root canals investigated (40/40). After instrumentation, all systems were associated with a highly significant reduction of the bacterial load and endotoxin levels, respectively: GI - Reciproc (99.34% and 91.69%); GII - Mtwo (99.86% and 83.11%); GIII - ProTaper (99.93% and 78.56%) and GIV - FKG Race(™) (99.99% and 82.52%) (P < 0.001). No statistical difference were found amongst the instrumentation systems regarding bacteria and endotoxin removal (P > 0.01). CONCLUSION: The reciprocating single file, Reciproc, was as effective as the multifile rotary systems for the removal of bacteria and endotoxins from root canals.

Susceptibility of as-yet-uncultivated and difficult-to-culture bacteria to chemomechanical procedures.

INTRODUCTION: A significant portion of the bacteria taking part of the microbiome associated with apical periodontitis still remain to be cultivated and phenotypically characterized. This molecular study evaluated the prevalence of selected as-yet-uncultivated and difficult-to-culture bacterial taxa in infected root canals and their susceptibility to chemomechanical procedures. METHODS: Root canals of single-rooted teeth with apical periodontitis were prepared using rotary nickel-titanium instruments and 2.5% sodium hypochlorite as the irrigant. DNA extracts from samples taken before (S1) and after (S2) chemomechanical preparation were surveyed for the presence of 7 as-yet-uncultivated phylotypes and 1 difficult-to-culture species using end-point polymerase chain reaction. Samples were also subjected to quantitative analysis of total bacteria and levels of the 2 most prevalent taxa. RESULTS: Bacteroidaceae sp. HOT-272 (24%) and Fretibacterium fastidiosum (20%) were the most prevalent taxa in S1. Their mean counts in S1 were 8.25 × 10(3) and 2.13 × 10(3) rRNA gene copies, corresponding to 0.18% and 0.55% of the total bacteria. Chemomechanical debridement promoted a highly statistically significant reduction in total bacterial counts (P < .001), but 64% of the canals were still positive for bacterial presence. Of the target taxa, only Bacteroidaceae sp. HOT-272 and F. fastidiosum were detected in S2 (each one in 1 sample). The reduction in counts of both taxa was also highly significant (P < .001). CONCLUSIONS: Findings confirmed that several as-yet-uncultivated and difficult-to-grow bacterial taxa can participate in the microbiome associated with apical periodontitis. Two of them were found in relatively high prevalence but rarely as a dominant species. Chemomechanical procedures were highly effective in completely eliminating these taxa or at least substantially reducing their numbers.

Clinical antibacterial effectiveness of the self-adjusting file system.

AIM: To evaluate in vivo the antibacterial effectiveness of the self-adjusting file (SAF) using molecular methods. METHODOLOGY: Root canals from single-rooted teeth with apical periodontitis were instrumented using the SAF system under continuous irrigation with 2.5% NaOCl. DNA extracts from samples taken before and after instrumentation were subjected to quantitative analysis of total bacteria counts and levels of streptococci by quantitative real-time polymerase chain reaction (qPCR). The reverse-capture checkerboard assay was also used to identify 28 bacterial taxa before (S1) and after (S2) SAF instrumentation. SAF was also compared with a conventional hand nickel-titanium instrumentation technique for total bacterial reduction. Data from qPCR were analysed statistically within groups using the Wilcoxon matched pairs test and between groups using the Mann-Whitney U-test and the Fisher's exact test, with significance level set at P < 0.05. RESULTS: Self-adjusting file significantly reduced the total bacterial counts from a mean number of 1.96 × 10(7) cells to 1.34 × 10(4) cells (P < 0.001). Quantitatively, the 99.9% reduction in total bacterial counts associated with the SAF system was significantly superior to the 95.1% reduction obtained by hand instrumentation (P < 0.001). Qualitatively, SAF resulted in significantly more cases with negative PCR results for bacteria (54.5%) than hand instrumentation (4.5%) (P < 0.001). The SAF system succeeded in significantly reducing the streptococcal levels, but four cases still harboured these bacteria in S2. Checkerboard analysis revealed that not only streptococci but also some anaerobic and even as-yet-uncultivated bacteria may resist the effects of chemomechanical procedures. CONCLUSION: The SAF instrumentation system was highly effective in reducing bacterial populations from infected root canals and performed significantly better than hand instrumentation. However, because half of the samples still had detectable bacteria after preparation with SAF, supplementary disinfection is still required to maximize bacterial elimination.

Correlative bacteriologic and micro-computed tomographic analysis of mandibular molar mesial canals prepared by self-adjusting file, reciproc, and twisted file systems.

INTRODUCTION: This ex vivo study evaluated the disinfecting and shaping ability of 3 protocols used in the preparation of mesial root canals of mandibular molars by means of correlative bacteriologic and micro-computed tomographic (µµCT) analysis. METHODS: The mesial canals of extracted mandibular molars were contaminated with Enterococcus faecalis for 30 days and assigned to 3 groups based on their anatomic configuration as determined by µCT analysis according to the preparation technique (Self-Adjusting File [ReDent-Nova, Ra'anana, Israel], Reciproc [VDW, Munich, Germany], and Twisted File [SybronEndo, Orange, CA]). In all groups, 2.5% NaOCl was the irrigant. Canal samples were taken before (S1) and after instrumentation (S2), and bacterial quantification was performed using culture. Next, mesial roots were subjected to additional µCT analysis in order to evaluate shaping of the canals. RESULTS: All instrumentation protocols promoted a highly significant intracanal bacterial reduction (P < .001). Intergroup quantitative and qualitative comparisons disclosed no significant differences between groups (P > .05). As for shaping, no statistical difference was observed between the techniques regarding the mean percentage of volume increase, the surface area increase, the unprepared surface area, and the relative unprepared surface area (P > .05). Correlative analysis showed no statistically significant relationship between bacterial reduction and the mean percentage increase of the analyzed parameters (P > .05). CONCLUSIONS: The 3 instrumentation systems have similar disinfecting and shaping performance in the preparation of mesial canals of mandibular molars.

Fate of non O157 Shiga toxigenic Escherichia coli in composted cattle manure

To determine the fate of Shiga toxigenic Escherichia coli (STEC) non-O 157 in composted manure from naturally colonized cattle, fresh manure was obtained from three cows carrying non-O157 STEC strains possessing the stx2 gene. Two composting systems were used: a 0.6m deep cave opened in the soil and an one meter high solid manure heap in a pyramidal architecture. Every day, for the 10 first days, and every five days for a month, one manure sample from three different points in both systems was collected and cultured to determine the presence of E. coli and the presence of the stx 2 gene in the cells. The temperature was verified at each sampling. STEC non-O157 E. coli cells survived for 8, 25 and 30 days at 42, 40 and 38ºC, respectively, in the deep cave and 4, 4 and 7 days at 65, 58 and 52ºC, respectively, in the heap, during the composting manure. Temperature and indigenous microorganisms appear to contribute to pathogen disappearance in the composting system. It is concluded that both composting systems were efficient to eliminate STEC cells. Land application of composted manure should minimize environmental risk associated with the dissemination of the pathogen

Determinou-se o tempo necessário para a eliminação de Escherichia coli Shigatoxigênica (STEC) não-O157 em esterco bovino composto, obtido de fezes frescas de três vacas portadoras de cepas STEC não-O157 que apresentavam o gene stx 2. Foram utilizados dois sistemas de compostagem, o primeiro foi um buraco de 0,6m escavado no solo e o segundo um monte apresentando uma arquitetura piramidal com um metro de altura. Todos os dias, durante os primeiros 10 dias e a cada cinco dias durante um mês, uma amostra de três pontos diferentes dos dois sistemas de compostagem foram coletadas e semeadas para determinar a presença de E. coli e a presença do gene stx 2 nas células, sendo que em cada coleta a temperatura do sistema de compostagem foi determinada. Células de STEC não-O157 sobreviveram por 8, 25 e 30 dias nas temperaturas de 42, 40 e 38ºC, respectivamente, no sistema enterrado no solo, enquanto que no sistema de monte as células foram detectadas por 4, 4 e 7 dias em temperaturas de 65, 58 e 52ºC, respectivamente. A temperatura e os microrganismos presentes na microbiota do sistema de compostagem parecem ser os responsáveis pela eliminação do patógeno. Pode-se concluir que os dois sistemas de compostagem utilizados mostraram-se eficientes na eliminação de células de STEC. A aplicação de esterco após compostagem deve diminuir o risco de contaminação ambiental e a disseminação do patógeno

Fate of non O157 Shiga toxigenic Escherichia coli in composted cattle manure / Eliminação de Escherichia coli Shigatoxigênica não-O157 em compostagem de esterco bovino

To determine the fate of Shiga toxigenic Escherichia coli (STEC) non-O 157 in composted manure from naturally colonized cattle, fresh manure was obtained from three cows carrying non-O157 STEC strains possessing the stx2 gene. Two composting systems were used: a 0.6m deep cave opened in the soil and an one meter high solid manure heap in a pyramidal architecture. Every day, for the 10 first days, and every five days for a month, one manure sample from three different points in both systems was collected and cultured to determine the presence of E. coli and the presence of the stx 2 gene in the cells. The temperature was verified at each sampling. STEC non-O157 E. coli cells survived for 8, 25 and 30 days at 42, 40 and 38ºC, respectively, in the deep cave and 4, 4 and 7 days at 65, 58 and 52ºC, respectively, in the heap, during the composting manure. Temperature and indigenous microorganisms appear to contribute to pathogen disappearance in the composting system. It is concluded that both composting systems were efficient to eliminate STEC cells. Land application of composted manure should minimize environmental risk associated with the dissemination of the pathogen (AU)

Determinou-se o tempo necessário para a eliminação de Escherichia coli Shigatoxigênica (STEC) não-O157 em esterco bovino composto, obtido de fezes frescas de três vacas portadoras de cepas STEC não-O157 que apresentavam o gene stx 2. Foram utilizados dois sistemas de compostagem, o primeiro foi um buraco de 0,6m escavado no solo e o segundo um monte apresentando uma arquitetura piramidal com um metro de altura. Todos os dias, durante os primeiros 10 dias e a cada cinco dias durante um mês, uma amostra de três pontos diferentes dos dois sistemas de compostagem foram coletadas e semeadas para determinar a presença de E. coli e a presença do gene stx 2 nas células, sendo que em cada coleta a temperatura do sistema de compostagem foi determinada. Células de STEC não-O157 sobreviveram por 8, 25 e 30 dias nas temperaturas de 42, 40 e 38ºC, respectivamente, no sistema enterrado no solo, enquanto que no sistema de monte as células foram detectadas por 4, 4 e 7 dias em temperaturas de 65, 58 e 52ºC, respectivamente. A temperatura e os microrganismos presentes na microbiota do sistema de compostagem parecem ser os responsáveis pela eliminação do patógeno. Pode-se concluir que os dois sistemas de compostagem utilizados mostraram-se eficientes na eliminação de células de STEC. A aplicação de esterco após compostagem deve diminuir o risco de contaminação ambiental e a disseminação do patógeno (AU)