NaOCl Solution Research Articles

Investigation of the Effect of Indirect Ultrasonic Force and Different Cements on the Bond Strength of Glass Fiber Posts in Teeth with Different Post Space Preparation Methods

(1) Background: The main aim of this study was to investigate the effects of different cements (Panavia V5, RelyX U200) and placement techniques (indirect ultrasonic vibration and manual method) on the bond strength of glass fiber posts in teeth with different post space preparation methods and to evaluate the failure types. (2) Methods: A total of 120 human upper central teeth were collected for the study. One week after root canal treatment, 10 mm long post space preparation was performed using post drills. Post spaces were irrigated in three different ways, namely NaOCl, NaOCl + EDTA solutions, and passive ultrasonic irrigation of NaOCl and EDTA solutions. Glass fiber posts were divided into two groups for resin cement systems, namely RelyX U200 (3M ESPE) or Panavia V5 (Kuraray Medical). Two different methods, namely the indirect ultrasonic vibration and the manual method, were applied for post placement. Then, 1.2 mm sections were taken from the middle and coronal regions of the inserted fiber posts for a push-out test. (3) Results: the Irrigation × Cement (p = 0.002), Cement × Section (p = 0.043) and Placement × Section (p = 0.049) pairwise interactions were statistically significant. (4) Conclusions: in our study, the effects of different post cavity preparations, different cement types, and different placement techniques on the bond strength of the fiber post were observed.

Three-Dimensional Planimetry Assessment of Dental Plaque-Covered Area Reduction after Rinsing with 0.2% Sodium Hypochlorite Solution as Part of a Guided Biofilm Therapy® Protocol-Pilot Longitudinal Study.

Background/Objectives: Less often employed as a rinsing solution for controlling oral biofilms, NaOCL was used in oral rinses at various concentrations in steps 1 and 4 of periodontal therapy. The aim of this study was to quantitatively evaluate the biofilm-disruptive properties of a 0.2% NaOCl solution in standardized oral rinses using dedicated plaque-disclosing agents and 3D scanning methods in patients undergoing the regular Guided Biofilm Therapy® protocol. Methods: Eight patients with at least 20 teeth present evenly distributed between the two arches were included. After 24 h of refraining from oral hygiene, dental arches were stained with a disclosing agent, the subjects rinsed for 20 s, clinical photographs and 3D scans were performed, subjects rinsed again for 20 s, photographs and 3D scans were performed again, and then the GBT® protocol was resumed as usual. Data representing areas covered with dental plaque were acquired using the "Medit Scan for Clinics" software and then underwent a post-processing and rendering process. The outcome variable was the percent reduction in the plaque-covered areas. Results: For the upper jaw, the estimated mean percent reduction in the biofilm-covered area was 39.65%, while for the mandible, it was 38.26%. The analysis of individual photographs revealed changes in the plaque-covered areas and reductions in the color intensity of the residual plaque-covered areas under identical lighting conditions. Conclusions: When analyzed using 3D intraoral scanning, the 0.2% NaOCl rinsing solution seems to be a clinically efficient disruptor/dissolvent of the oral biofilm, both when integrated into modern protocols of periodontal therapy like GBT® and for home self-care.

The treatment of Enterococcus faecalis related root canal biofilms with phage therapy

As one of the global concerns, antimicrobial resistance crisis increases the clinical importance of Enterococcus species. Enterococcus faecalis (E. faecalis) specifically penetrates the dentinal tubules and remains prevalent even after endodontic treatment. It has also biofilm forming character as well as the development of resistance to antibiotics. Sodium hypochlorite (NaOCl) is considered the gold standard among antibacterial washing solutions. However, due to its toxic effects, its usage limitations have led to the search for natural, non-toxic alternatives. Phages can be considered an important alternative because of their effects on specific bacteria. The objective of this study was to compare the effect of isolated active vB_Ef1 phage on the removal of E. faecalis biofilm in dentin, together and separately with the chemical irrigation solution NaOCl. As a result of study, the optimal NaOCl solution concentration to be applied with vB_Ef1 phage is 0.5 %, and the use of solution at this value reduces the biofilm mass by 84 %, reaching the highest biofilm mass reduction value. It was found that the combination of phage and NaOCl at appropriate concentrations had the strongest biofilm disruption effect.

Testing the Inhibitory Activity of Bajakah Tampala (Spatholobus littoralis Hassk) Wood Extract from South Kalimantan on the Growth of Pseudomonas aeruginosa Bacteria

Background: Dental root canal treatment aims to eliminate microorganisms that cause infection in the tooth root canal, one of which is Pseudomonas aeruginosa bacteria. Disinfection of dental root canals with 0.5% - 5.25% NaOCl irrigation solution is still constrained so it is necessary to look for alternatives. The bajakah tampala plant (Spatholubus littoralis Hassk) is known to contain antibacterial compounds, namely flavonoids, saponins, tannins, and polyphenols. Objective: To evaluate the inhibitory activity of bajakah tampala wood extract (Spatholobus littoralis Hassk) from South Kalimantan against the growth of Pseudomonas aeruginosa bacteria. Method: Post test-only with control group design. The test groups were 100%, 75%, 50% bajakah tampala wood extract solution and positive control 2.5% NaOCL solution. Extraction using maceration method. Bacterial growth inhibition was tested by well diffusion method. Inhibition zone was measured using a caliper. Results: The growth inhibition from the largest to the smallest was shown by 75%, 100% and 50% bajakah wood extracts with mean inhibition zone diameters of 5.05 mm, 3.84 mm and 2.02 mm respectively, meanwhile the positive control was 16.14 mm. One Way Anova test showed significant difference in inhibition (p<0.05) in all groups. Post hoc test showed no significant difference in the inhibition of 75% bajakah tampala wood extract with 100% extract. The inhibition of 100% and 75% extracts was significantly greater than 50% extract. The inhibition of NaOCL 2.5% was significantly greater than all groups of test extracts. Conclusion: Bajakah tampala wood extract has inhibitory activity against the growth of Pseudomonas aeruginosa bacteria.

First Report of Stem Blight of Mungbean (Vigna radiata) Caused by Diaporthe longicolla in the United States.

Mungbean (Vigna radiata) is primarily grown in Asia and directly consumed by humans. U.S. consumers embraced mungbean as a plant-based protein in vegan eggs and meat substitutes. New cultivars are being developed for American farmers because of the crop's tolerance to heat and drought, and its adaptability to current farming infrastructure. Mungbean's short season complements various cropping systems such as intercropping, alternative cropping, and green manure. With rotations and inclusion with soybean systems, there is a concern about the overlap of common pathogens for soybean and mungbean. During August 2022 when mungbeans reached full maturity (growth stage R6), reddish-brown and necrotic stem lesions with linear rows of black pycnidia were observed on Berken and OK2000 cultivars at fields located in Hancock County, IA and Story County, IA in the United States. Pycnidia measured 0.5-0.6mm in length. Disease incidence was approximately 10% of plants in Hancock County, IA and less than 3% of plants in Story County, IA. Pycnidia from 16 plants were excised and immersed in a 0.5% NaOCl solution for 1 min, rinsed with autoclaved distilled water, and placed onto potato dextrose agar (PDA). Eighteen isolates were hyphal tipped and grown on PDA and were stored at 25°C. Isolates were then visually identified by culture and conidia morphology (Hobbs et al. 1985, Santos et al. 2011). Colonies were cream to white, dense, and floccose. Large black stromata were formed in a concentric pattern or scattered; alpha conidia were ellipsoidal. Template DNA for PCR amplification of the internal transcribed spacer region of the nuclear ribosomal DNA operon (ITS) and the beta-tubulin gene (TUB) was extracted from 18 isolates by scraping mycelia with a sterile pipette tip and transferring it into 50 ul of PrepMan Ultra Sample Reagent (Applied Biosystems, Foster City, California, USA). Fungal primers were ITS1 and ITS4 (White et al. 1990) and Bt-2F/Bt-2R (Udayanga et al. 2014). Sequences of isolates obtained from fields in both counties were identical, providing no species diversity. GenBank accession numbers for the ITS region were PP105598 and PP105599; PP108254 and PP108255 for TUB sequences. BLAST results showed the ITS 550/550 base pairs with type specimen D. longicolla ATCC 60326 GB NR_144924 and the TUB 446/446 base pairs with type specimen D. longicolla ATCC 60325 GB KJ610883. Thus, the isolates were identified as Diaporthe longicolla (Hobbs) J.M. Santos, Vrandecic & A.J.L. Phillips based on morphology and molecular characters (Santo et al 2011; Udayanga et al. 2014). To confirm the pathogenicity of the D. longicolla isolates, twenty mungbean plants (cv. Berken and OK2000) were grown in the greenhouse at 85% RH and 16hr light for 20 days. Inoculum was prepared by placing sterile toothpicks on 1/3 PDA with a single representative isolate from each field location for 21 days (Ghimire et al. 2019). Mungbean plants were grown in a 10cm-by-10cm pot containing a greenhouse professional growing mix (Sungrow, Agawam, Massachusetts, USA) and grown for 30 days post emergence. After 12 days of growing, a 3mm segment of the infested toothpick was inserted into a stem wound below the first trifoliate and sealed with parafilm. A sterile toothpick was inserted into the control plants. After 14 days, red lesions extended downward 1 to 3 cm from the inoculation site, and white mycelial was present in the wound. At 21 days red lesions spanned 3 to 9 cm upward and downward from the inoculation site. Pycnidia were present on collapsed stem tissue, and leaves became chlorotic. Damage was limited to 2mm from the mock-inoculation site, with no discoloration in the control plants. Symptomatic tissues were plated and compared to the original isolates. Alpha conidia were ellipsoidal with the base end rounded. To our knowledge, this is the first report of Diaporthe longicolla causing disease on mungbean within the U.S. and worldwide. The presence of this disease in two locations suggests the potential for Diaporthe longicolla to be a serious disease of mungbean in the future.

Do the apical limit and diameter of the instrumentation influence the extrusion of sodium hypochlorite solution? A micro-CT study.

This study evaluated the influence of the apical limit and the diameter of the instrumentation on the extrusion of the irrigating solution. Sixty human lower premolars were distributed into 5 groups according to the apical limit and diameter of the root canal instrumentation (n = 12): (G-140) R40 instrument 1mm above the tooth length (TL) (Control); (G-150) R50 instrument 1mm above the TL; (GF25) R25 instrument at the TL; (GF40) R40 instrument at the root canal length and (GF50) R50 instrument at the TL. A space of 33.5 ± 1 mm3 was created for storage of the extruded solution. Omnipaque contrast was added to the NaOCl solution in a 1:2 ratio, totaling 15mL of solution to allow its visualization when extruded in microcomputed tomography. Final irrigation was performed with a syringe and needle and activated with a passive ultrasonic irrigation (PUI). Three activation cycles were performed. Data were tabulated and statistically analyzed (one-way ANOVA) with a significance level set at 5%. It was observed irrigating solution extrusion after final irrigation in all groups. However, there were no statistically significant differences among experimental groups, regardless of the diameter or apical limit of root canal instrumentation (p > 0.05). The different apical limits and diameters of root canal instrumentation did not influence the volume of extruded irrigating solution using passive ultrasonic irrigation.

Comparative evaluation of continuous and sequential chelation on the dentinal tubule penetration of bioceramic-based sealer - A confocal laser scanning microscopic study.

Effective endodontic treatment relies heavily on proper instrumentation, thorough disinfection, and precise three-dimensional obturation of the root canal system. This study aims to evaluate and compare continuous chelation (etidronic acid and sodium hypochlorite [NaOCl]) and sequential chelation (citric acid and NaOCl) on the dentinal tubule penetration of bioceramic-based sealer (Bio-C)-A confocal laser scanning microscopic study. In vitro experimental study. Sixty extracted permanent single-rooted teeth were selected and decoronated below cementoenamel junction to get a standardized length of 12 mm across all samples followed by enlargement of root canals till rotary ProTaper F3. The samples were randomly divided into three groups: Group I (control): Canals were irrigated with 3 mL 17% ethylenediaminetetraacetic acid/3 mL 3% NaOCl; Group II: Canals were irrigated with 10 mL etidronic acid + NaOCl solution; and Group III: Canals were irrigated with 3 mL 10% citric acid followed by 3 mL 3% NaOCl. 0.1% rhodamine B dye was mixed with Bio-C sealer followed by obturation. All the samples were subjected to confocal laser scanning microscopy evaluation. One-way analysis of variance with the Bonferroni post hoc test was used for the statistical analysis. The level of significance was set at P < 0.05. The pairwise comparison of depth of penetration showed statistically significant results in all the coronal, middle, and apical thirds. Group II showed better depth of penetration than Groups I and III. Continuous chelation protocol using etidronic acid and NaOCl showed greater and statistically significant sealer penetration depth when compared to the sequential chelation protocol, i.e., citric acid at all three levels.

The Optimization of Aerobic Bacteria Inactivation in Tilapia (Oreochromis niloticus) Fillets using Micro-Nano Bubbles of Carbon Dioxide and Shelf-Life Extension

This study aims to examine the influence of NaOCl, NaCl and contact time on the inactivation of aerobic bacteria in tilapia fillets treated with micro-nano bubbles of CO2 (CO2 MNB) in a washing process of fish fillets, and compared to soaking with tap water and untreated fillets for their shelf-life extensions. Response surface methodology (RSM) with a central composite design was used to compare and predict of the inactivation effects. The fish fillets were soaked in a NaOCl solution before washing with a NaCl solution and CO2 MNB produced from an MNB generator system, maintaining the liquid temperature in the range of 4–7 °C for all experiments. According to the regression analysis from the experimental design, aerobic bacteria inactivation was reduced by 1.509 log CFU/g at 100 mg/L NaOCl, 10%w/v NaCl, and a contact time of 32 min with CO2 MNB. The experimental value of the reduction of aerobic bacteria by 1.503 ± 0.009 log CFU/g (before washing 5.623 log CFU/g; after washing 4.120 log CFU/g) was found after treatment under the aforementioned condition. The number of aerobic bacteria counted on the tilapia fillets treated with the upper condition after being stored at 4 ± 2 °C for 7 d was below the acceptable limits, but untreated and treated with tap water had bacteria counts exceeding the upper microbial limit (6 log CFU/g). The combined results showed that the NaOCl, NaCl solution and CO2 MNB treatment could extend the storage time by more than 7 d.

The effect of ascorbic acid, QMix and other conditioning solutions on the bond strength of adhesively luted glass fiber-reinforced composite posts to root dentin, a laboratory study

PurposeThis study was designed to investigate the effect of post space conditioning with different solutions on the bond strength of glass FRC posts and evaluate the mode of resin failure. MethodsSixty extracted human single rooted teeth were root filled using a resin sealer and suitable gutta-percha cones. Post spaces were prepared, and the teeth randomly allocated into 5 groups according to their irrigation regimens which included the following solutions: 17 % EDTA, 2 % CHX, 3 % NaOCl, 10 % ascorbic acid and QMix solutions. Post spaces were irrigated with 5 ml of the solution for 15 s and subsequently washed with distilled water and dried with paper points. Glass FRC posts were cemented into their spaces using a self-adhesive resin cement. The specimens were mounted in plexiglass molds using autopolymerizing acrylic resin. A universal testing machine was used to measure post retention at a crosshead speed of 2 mm/min. The results were analyzed by one-way ANOVA followed by Tukey HSD test (α = 0.05). Dislodged posts and post spaces were examined microscopically to evaluate retention failure. ResultsThe Ascorbic acid group exhibited the highest mean retentive strength value at 229 N, followed by QMix at 198 N, NaOCl at 186 N, CHX at 170 N, and EDTA at 124 N. The mean value of the ascorbic acid group was significantly higher than EDTA group, p = 0.012. The failure category was primarily mixed. ConclusionsRinsing post spaces with ascorbic acid exhibited significantly superior bond strength. The failure mode was mixed. Clinical significanceIrrigating post spaces with ascorbic acid solution before luting FRC posts significantly improves their bond strength compared to irrigation with EDTA solution. Irrigation with QMix solution produced the second highest retentive strength but showed no statistical significance when compared to using ascorbic acid, NaOCl, CHX, or EDTA solutions.

Cytotoxic Effect of Two Different Concentrations of Sodium Hypochlorite: An In-Vitro Study.

Introduction The endodontic treatment of primary teeth presents considerable complications due to their distinct anatomical properties. In order to achieve a successful endodontic treatment, certain factors must be assessed. These factors include a precise diagnosis, thorough cleaning, and a reliable disinfection protocol. Although sodium hypochlorite (NaOCl) has been effective as an irrigation agent in primary teeth, it is important to recognize that higher concentrations of NaOCl might possibly inflict toxic harm on the periapical environment if they penetrate the tooth's apical foramina. Since primary teeth are important, pediatric dentists must choose an appropriate NaOCl concentration for root canal irrigation, as higher concentrations can be toxic. Thus, the current investigation examined the cytotoxicity of two different NaOCl concentrations at various volumes. Methods To evaluate the cytotoxicity potential, a culture ofnauplii (brine shrimp) was prepared and subjected to testing. For the test, 5, 10, 20, and 40 µL of 1% and 3% NaOCl were added to the brine shrimp culture at different concentrations, and saline was used as a control. After a span of 24 hours, the total number of alive nauplii was duly noted. Results After 24 hours, nauplii showed no mortality in the control group. For 1% NaOCl, mortality ranged from 10% to 20% across volumes, with no significant differences (p = 0.193). In contrast, 3% NaOCl caused significantly higher mortality: 20% at 5 µL, 30% at 10 and 20 µL, and 60% at 40 µL (p = 0.007). Tukey's analysis revealed no significant differences for 1% NaOCl (p > 0.05) but significant differences for 3% NaOCl at 40 µL (p < 0.05). Conclusion Based on the results of the present study, it was observed that a 1% NaOCl solution exhibited a lower level of toxicity in comparison to a 3% NaOCl solution. These findings highlight the importance of using lower concentrations of NaOCl for endodontic irrigation in pediatric dentistry to reduce the risk of tissue damage and ensure safer outcomes for young patients.

Compatibility of Dual Rinse 1-Hydroxyethane-1,1-Diphosphonic Acid with Sodium Hypochlorite Solutions Containing Proprietary Additives

IntroductionA salt of 1-hydroxyethylidene-1,1-diphosphonic acid (HEDP) was commercialized some years ago. This so-called Dual Rinse (DR) HEDP has been tested for its combined actions and interactions with pure sodium hypochlorite (NaOCl) solutions. The aim of this study was to assess DR interactions with NaOCl solutions containing proprietary additives. MethodsDesignated endodontic NaOCl solutions that come at 2 different strengths (2% and 5.25%), and without or with additives (Chloraxid and Chloraxid Extra, respectively) were tested. Some of their key chemical and physical properties were determined in the presence or absence of HEDP (9%) in the solution. In addition, soft tissue dissolution efficacy of test and control solutions was assessed in extracted teeth (n = 10 per group, 90 teeth in total). ResultsThe NaOCl content in the 2% Chloraxid solutions was slightly above, that in the 5.25% solutions below the labeled concentration. The additives in the Chloraxid Extra solutions neither altered their surface tension nor their viscosity. The addition of HEDP to any of the tested NaOCl solutions reduced their pH, and increased their surface tension and viscosity. HEDP-induced available chlorine loss over the first hour was similar between all NaOCl solutions. Soft tissue dissolution in the root canals was affected by NaOCl concentration, but neither by any proprietary NaOCl additive in the Chloraxid solutions, nor the addition of DR HEDP. ConclusionsProprietary additives had no influence on any of the tested parameters, including surface tension of the "Extra" solutions. The increased viscosity of combined NaOCl/HEDP solutions deserves further attention.

Occurrence of Fusarium Crown Rot Caused by Fusarium culmorum on Winter Wheat in Xinjiang Uygur Autonomous Region, China.

Wheat (Triticum aestivum L.) is the predominant grain crop and plays a pivotal role in grain production in Xinjiang Uygur Autonomous Region (XUAR), China. Its cultivated area constitutes approximately half of the total sown area of grain crops in XUAR, with 1.14 million hectares in 2021. Fusarium crown rot (FCR) of wheat, caused by Fusarium culmorum (W.G. Smith) Sacc., is one of the most devastating soil-borne diseases known to seriously reduce grain yield (Ma et al. 2024; Saad et al. 2023). In 2016, FCR of wheat, caused by F. culmorum, was firstly identified in Henan Province, China (Li et al. 2016). In June 2023, during the investigation of FCR of wheat in Aksu Prefecture, XUAR, FCR on winter wheat (cv. Xindong 20) was found (82.761349°E, 41.612202°N). The grain-filling period for winter wheat in early June coincided with a period of high temperatures and water demand in Aksu Prefecture. Approximately 8% of the Xindong 20 wheat plants exhibited symptoms of white heads and browning at the stem base, with the disease present in 82% of the wheat fields surveyed. To identify the pathogens, 20 samples of diseased stem basal tissue, each 0.5 cm in length, were collected and sterilized with 75% alcohol for 30s and 5% NaOCl solution for 2 min, followed by three rinses with sterile water. These samples were then plated onto potato dextrose agar (PDA) medium at 25°C for 5 days. A total of 17 isolates with consistent morphological characteristics were obtained using single-spore technique, with an isolation rate of 85%. The isolated strains exhibited rapid growth on PDA, producing fluffy, pale-yellow hyphae, and accumulating a pale-yellow to dark red pigment on the bottom of the medium. On carnation leaf agar (CLA), these strains formed orange colonies due to the aggregation of a large number of macroconidia. The macroconidia were short and thick, with three to four septa and rounded apical cell, averaging 31.94 to 40.96 × 5.62 to 6.71 μm (Magnification of ×400). Microconidia were not observed. These morphological characters were consistent with those of F. culmorum (Leslie and Summerell. 2006). Two isolates (D-9 and D-11) were selected for molecular identification. The EF-1α gene fragment was amplified using primers EF1/EF2 (5'-ATGGGTAAGGARGACAAGAC-3'/5'-GGARGTACCAGTSATCATG-3') as previously described by O'Donnell et al. (1998). The two 665 bp PCR products were sequenced and submitted to GenBank (GenBank Accession No: PP763247 and PP763248) with 99. 7% identity to the published F. culmorum sequences (e.g., OP985478, OP985477, MG195126, KX702638). The molecular identification was further confirmed by F. culmorum species-specific PCR primers FcOIF/FcOIR (Nicholson et al. 1998). The expected PCR products of 553 bp were produced only in F. culmorum. Strains D-9 and D-11 were used to conduct the pathogenicity experiment on 7-day-old winter wheat (cv. Xindong 20) using drip in the lower stem inoculation method with a 10-μl of 106 macroconidia ml-1 suspension, and the control 7-day-old winter wheat were treated with sterile water (Xu et al. 2017). The experiments were replicated five times in a greenhouse at temperatures ranging from 20℃ to 25℃. After 4 weeks, all inoculated wheat seedlings showed stem base browning or even death. No symptoms were observed on the control plants. The fungus was reisolated from all inoculated wheat plants by the method described above and identified by morphological and PCR amplification using F. culmorum species-specific primers FcOIF/FcOIR. No F. culmorum was isolated from the control wheat plants, fulfilling Koch's postulates. To the best of our knowledge, this is the first report of F.culmorum causing FCR on winter wheat in XUAR, China. Considering wheat is the predominant grain crop and plays a pivotal role in grain production in China, necessary measures should be taken to prevent the spread of F. culmorum to other regions.

First Report of Pantoea ananatis causing leaf streak disease on wheat (Triticum aestivum) in the United States of America.

Wheat (Triticum aestivum) loses 21.5% yield to pests and diseases annually (Savary et al. 2019). Among the wheat diseases, bacterial leaf streak (BLS) is a growing problem, costing $78.5 million in losses (https://cropprotectionnetwork.org/). In July 2022, we sampled winter wheat leaf samples at Volga (44.30, -96.92), South Dakota, USA with an estimated disease incidence of 40% (n=100). The typical symptoms were water-soaking with large necrotic and chlorotic streaks extending the length of the leaves and were strikingly similar to BLS. To isolate the pathogen, leaves were cut lengthwise into 1 cm pieces and surface-sterilized using a 10% NaOCl solution for 3 min, followed by 70% ethanol for 3 min, and then rinsed with sterile distilled water and placed in 500 ul of sterile distilled water for 5 min and using a sterile loop the water was streaked over a plate of Nutrient Agar (NA). Following Duveiller et al. (1997), the streaked plate was incubated in the dark at 28℃ for 48 h. Observed single colonies were sub-cultured thrice onto fresh NA plates to obtain a pure culture. We named the culture SD101. Bacteria were found to be gram-negative with a colony morphology initially raised, smooth, and white that later turned yellow. DNA was extracted using the Wizard HMW DNA Extraction Kit (Promega, Madison, WI) following the manufacturer's protocol, and sequenced using Nanopore MinION R9.4 (Oxford Nanopore Technology). We used the Rapid Annotation Using Subsystems Technology approach (Aziz eal. 2008) to extract the 16S rDNA, DNA gyrase subunit B (gyrB), and translation initiation factor IF-2 (infB) gene sequences that were deposited in GenBank under accession numbers PP329908.1 for 16S rDNA, PP496481 for infB, and PP328920.1 for gyrB. Homology analysis using CLC Genomics Workbench 22.0.2 (QIAGEN) and BLASTn against the GenBank nucleotide database resulted in a 99.74% match (1543/ 1547 bp) of the 16S sequence, 99.59% match (2674/ 2685 bp) of the infB sequence, and 99.42% match (2396/ 2410 bp) of the gyrB sequence with Pantoea ananatis strain AJ13355 (AP012032). To test pathogenicity, seeds of spring wheat breeding line SD4892 were planted in 30 cm × 30 cm pots in a greenhouse under a 16 h light photoperiod. The inoculum was prepared from 48-h-old NA plates of SD101 rinsed with 1X Phosphate Buffer Saline (PBS buffer), adjusted to an OD600 = 1.0, and amended with two drops of Tween 20 (polyoxyethylene sorbitol ester, Millipore Sigma). PBS with Tween 20 was used as a negative control. The inoculum was sprayed on 15 replicates of 15-day-old seedlings, kept at 95% relative humidity for 48 h, then moved to the greenhouse at 23 to 25°C. The symptoms appeared as water soaking that later turned to necrotic streaks with surrounding chlorosis on all 15 inoculated plants while control plants remained healthy. The pathogen was re-isolated from the leaves as described above. The 16S rDNA, infB, and gyrB sequences amplified and sequenced were identical to the gene sequences from the whole genome sequencing. The experiment was repeated with the same results, completing Koch's postulates. Although P. ananatis is pathogenic on corn, rice, and other plant species in the United States (Coutinho et al. 2009), and was reported pathogenic on wheat in Poland (Krawczyk et al. 2020), this is the first report of its pathogenicity on wheat in the United States. The prevalence, and incidence of BLS disease caused by P. ananatis in wheat is needed to estimate its threat to wheat production and to develop management strategies.

First Report of Exserohilum rostratum Causing Brown Spot on Spartina alterniflora in China.

Spartina alterniflora Loisel, a perennial grass, has become an invasive species in China's coastal wetlands (Zhang et al. 2018). In July 2021, brown spot symptoms were observed on S. alterniflora in a coastal wetland (21°45'48″N, 108°44'00″E) in Beihai City, Guangxi Province, China. The disease affected approximately 50% of the plants in the surveyed area (0.2 ha) and was also observed in other regions of Beihai. It caused brown lesions with a gray or whitish center on the leaves and stems of S. alterniflora. As the disease developed, it ultimately led to leaf shedding and plant death. To isolate the causal agent, 18 fragments (~ 5 mm) from six symptomatic plants (3 leaf pieces per plant) were surface sterilized with 1% NaOCl solution for 2 min and rinsed three times with sterilized water. Subsequently, the tissues were placed on potato dextrose agar (PDA) medium supplemented with chloramphenicol (0.1 g/liter) and incubated at 28°C for three days. The hyphal tips were transferred onto fresh PDA to obtain pure cultures. A total of 25 isolates were obtained, 20 of which shared similar morphologies, while the remaining five exhibited distinct morphological characteristics and were non-pathogenic to S. alterniflora. Three isolates (MC16.1.3, MC16.6.2, and MC16.8.3) were randomly selected from the 20 for further investigation. The colonies on PDA were flat with dense aerial mycelia. The colony margins were entire, light brown in the centre, white to grey at the margin; reverse dark brown in the centre, gray at the margin. Conidia were straight to slightly curved, light olive-brown to dark olive-brown, septate, measured 33.5 to 79.1 μm × 10.4 to 18.7 μm (average 52.9 × 14.4 μm, n = 100), with a distinctly protruding hilum swelled from the basal cell. For molecular identification, the genomic DNA was extracted from mycelium on PDA using the CTAB method (Guo et al. 2000). The internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation elongation factor 1 alpha (TEF1-α) genes were amplified and sequenced with the primer pairs ITS1/ITS4 (White et al. 1990), GPD1/GPD2 (Berbee et al. 1999), and EF1-983/EF1-2218 (Rehner et al. 2005), respectively. A BLAST analysis revealed that the ITS (OR516787-9), GAPDH (OR523686-8), and TEF-α (OR523683-5) had 99.1 to 99.7% identity with those of E. rostratum strains BRIP 11417 (LT837836, LT882553, and LT896656) and CBS 128061 (KT265240, LT715900, and LT896658) (Hernández-Restrepo et al. 2018). Based on the concatenated sequences, a phylogenetic tree generated by PhyloSuite software (Zhang et al., 2020) through Bayesian inference (BI) and Maximum Likelihood (ML) methods placed the isolates within E. rostratum. These morphological characteristics and molecular analyses confirmed the pathogen as E. rostratum (Hernández-Restrepo et al. 2018; Kaboré et al. 2022). To confirm pathogenicity, a conidial water suspension (~ 1 × 106 conidia/ml) of each of the three strains was inoculated on nine healthy S. alterniflora plants that had been grown for six months. Control plants were treated with sterile water. All plants were then enclosed in plastic bags and incubated in a greenhouse at 28°C. Six days after inoculation, the plants exhibited symptoms similar to those observed in nature. The control plants developed no symptoms. These experiments were replicated three times with similar results. To fulfill Koch's postulates, E. rostratum was consistently re-isolated from symptomatic tissue and confirmed by morphology and sequencing, whereas no fungus was isolated from the control plants. In recent years, S. alterniflora has posed a serious threat to the indigenous biodiversity of wetland ecosystems (Zhang et al. 2018). To our knowledge, this is the first report of E. rostratum causing brown spot on S. alterniflora worldwide.

Comparative evaluation of the efficacy of 1% and 3% Sodium hypochlorite in reducing the microbial counts in primary teeth root canals using Bioluminometer – A randomized clinical trial

PurposeSodium hypochlorite (NaOCl) is commonly used to irrigate primary teeth during pulpectomy. Although high concentrations of NaOCl are effective, they pose a risk of toxic damage to periapical tissues if NaOCl penetrates through the apical foramen. Therefore, low concentrations of NaOCl are preferred to mitigate this risk. However, concerns persist regarding the antibacterial efficacy of low concentrations of NaOCl compared to high concentrations. The objective of this study was to assess and compare the efficacy of 1% and 3% NaOCl irrigation in reducing bacterial load within primary teeth root canals. Materials and methodsThis clinical study involved forty participants divided into two groups. Group 1 (n = 20) received canal irrigation with 1 % NaOCl solution, while Group 2 (n = 20) received canal irrigation with 3 % NaOCl solution. Microbial samples were collected from the root canal using a paper point before and after irrigation. The samples were aseptically transferred to ultra-snap tubes and then analyzed using a Bioluminometer. The results were recorded. ResultsBoth groups exhibited a decrease in bacterial count after irrigation. The mean colony count post irrigation for 3 % NaOCl was 258.05 ± 28.61, and for 1 % NaOCl it was 267.60 ± 30.56. However, no statistically significant difference was observed upon intergroup comparison. ConclusionThis study shows that 1% NaOCl is equally effective as 3% NaOCl in reducing bacterial count in root canals. Thus, using 1% NaOCl as an irrigant is appropriate in clinical practice.

The effect of bioactive glass-based, bioceramic based and epoxy amine resin based root canal sealers on post-obturation pain: A double blinded randomized controlled trial.

The aim of this study was to compare the effect of two calcium silicate-based and an epoxy resin-based root canal sealers on postoperative pain and analgesic intake following single-visit root canal treatment. Ninety patients with at least one first or second molar tooth diagnosed as symptomatic irreversible pulpitis and symptomatic apical periodontitis were selected and allocated into three groups (n=30) according to the sealer used. Root canals were prepared using Protaper Gold instruments (Dentsply Sirona) in a crown down technique and irrigated with 2.5% NaOCl (Calyx, India) and saline solution. Root canal filling was then accomplished with a single cone obturation technique and treated in a single visit by the same endodontist. Patients were told to use a Visual Analog Scale (VAS) to rate their postoperative pain severity as none, minimal, moderate, or severe after 6 h, 24 h, 48 h, 5 days and 7 days following obturation using the appropriate sealers. The need for analgesic intake was also recorded. The data were statistically analyzed. Results showed a significant difference among the studied groups. Bio-C Sealer Ion+ reported the least pain score followed by Nishika Canal Sealer BG and AH plus sealer at all the time intervals recorded. The intergroup analysis, revealed was a significant difference in postoperative pain at 6 h (p=0.000) and 24 h (p = 0.028), but not at 48 h, 5 day or 7 days (P > 0.05). VAS ratings for all the three groups decreased over time. Also, there were significant differences between the means of analgesic intake among 3 groups (p=0.022). Analgesic intake in group BIO-C Sealer Ion+ is significantly lesser than AH Plus and Nishika Canal Sealer BG group. Calcium silicate-based sealer (Nishika Canal Sealer BG and Bio-C Sealer Ion+) resulted in significantly lower levels of pain as compared to epoxy resin-based sealer (AH Plus) at 6h and 24-h interval, there was no significant difference in postoperative pain occurrence at 48-h, 5 day and 7-day period. The analgesic intake in Bio-C Sealer Ion+ group is significantly lesser than Nishika Canal Sealer BG and AH Plus group.

Discovery of Fusarium pseudograminearum Causing Crown Rot of Winter Wheat in Xinjiang Uygur Autonomous Region, China.

Fusarium pseudograminearum is an important plant pathogen that invades many crops (Zhang et al. 2018). Since it was first discovered in Australia in 1951, F. pseudograminearum has been reported in many countries and regions and caused huge economic losses (Burgess et al. 2001). In 2012, crown rot of wheat caused by F. pseudograminearum was discovered for the first time in Henan Province, China (Li et al. 2012). Wheat (Triticum aestivum L.) is one of the most important food crops in Xinjiang Uygur Autonomous Region (XUAR), with 1.07 million hectares cultivated in 2020. In June 2023, a survey of crown rot disease was carried out in winter wheat cv. Xindong 20 in Hotan area, XUAR, China (80.148907°E, 37.051474°N). About 5% of wheat plants showed symptoms of crown rot such as browning of the stem base and white head. The disease was observed in 85% of wheat fields. In order to identify the pathogens, 36 pieces of diseased stem basal tissue, 0.5 cm in length, were collected and sterilized with 75% alcohol for 30s and 5% NaOCl solution for 2 min, then rinsed three times with sterile water and placed on potato dextrose agar (PDA) medium at 25°C. A total of 27 isolates with consistent morphological characteristics were obtained using single-spore technique (Leslie and Summerell. 2006), and the isolation rate was 75%. The isolates grew rapidly on PDA, produced large numbers of fluffy white hyphae, and pink pigment accumulated in the medium. The isolates were grown on 2% mung bean flour medium and identified by morphological and molecular methods. Macroconidia were abundant, relatively slender, curved to almost straight, commonly two to seven septate, and averaged 22 to 72 × 1.8 to 4.9 μm. Microconidia were not observed. The morphological characters are consistent with Fusarium (Aoki and O'Donnell. 1999). Two isolates (LP-1 and LP-3) were selected for molecular identification. Primers EF1/EF2 (5'-ATGGGTAAGGARGACAAGAC-3'/5'-GGARGTACCAGTSATCATG-3') were used to amplify a portion of the EF-1α gene (O'Donnell et al. 1998). The two 696 bp PCR products were sequenced and submitted to GenBank. The EF-1α gene sequences (GenBank Accession No: PP062794 and PP062795) shared 99.9% identity (695/696) with published F.pseudograminearum sequences (e.g., OP105187, OP105184, OP105179, OP105173). The identification was further confirmed by F. pseudograminearum species-specific PCR primers Fp1-1/Fp1-2 (Aoki and O'Donnell. 1999). The expected PCR products of 518 bp were produced only in F. pseudograminearum. Pathogenicity tests of LP-1 and LP-3 isolates were performed on 7-day-old seedlings of winter wheat cv. Xindong 20 using the drip inoculation method with a 10-μl of a 106 macroconidia ml-1 suspension near the stem base (Xu et al. 2017). The experiment was repeated five times in a 20 to 25°C greenhouse. Control seedlings were treated with sterile water. After 4 weeks, wheat seedling death and crown browning occurred in the inoculated plants with over 90% incidence. No symptoms were observed in the control plants. The pathogen was reisolated from the inoculated plants by the method described above and identified by morphological and PCR amplification using F. pseudograminearum species-specific primers Fp1-1/Fp1-2. No F. pseudograminearum was isolated from the control plants, fulfilling Koch's postulates. To our knowledge, this is the first report of F. pseudograminearum causing crown rot of winter wheat in XUAR of China. Since F. pseudograminearum can cause great damage to wheat, one of the most important food crops in China, necessary measures should be taken to prevent the spread of F. pseudograminearum to other regions.

Interleukin-8 (IL-8) levels in gingival crevicular fluid during root canal treatment of molar teeth with symptomatic irreversible pulpitis: impact of varying sodium hypochlorite concentrations

BackgroundThe aim of this study was to evaluate the effect of the use of different NaOCl concentrations (1%, 2.5%, and 5.25%) during root canal treatment of molar teeth with symptomatic irreversible pulpitis on the change of the IL-8 level in gingival crevicular fluid (GFC).MethodsGCF sampling was performed on experimental tooth with irreversible pulpitis before and after treatment and also contralateral healthy tooth of 54 patients. The patients were divided into three groups according to concentration of NaOCl solution (n = 18); 1%, 2.5%, and 5.25% NaOCl solution. GCF sampling from experimental teeth was repeated one week after root canal treatment. Statistical analysis was performed using Mann-Whitney U, Wilcoxon test, one-way ANOVA and Pearson correlation analysis.ResultsThere was a significant correlation between IL-8 levels in GCF samples taken from teeth with pulpitis before treatment and from healthy contralateral teeth (p = .000). Furthermore, the pretreatment IL-8 level was significantly higher than the posttreatment IL-8 level(p < .05). The effect of the NaOCl concentration on the change in IL-8 level betweeen pre and post treatment was not found statistically significant (p > .05).ConclusionsThe use of NaOCl during root canal treatment can effectively reduce the levels of IL-8 in GCF and improve clinical outcomes.Trial registrationThis study was registred in the Institutional Review Board and the Ethics Committee of the University (No:11) on 15/12/2021.

An invitro comparison of antimicrobial efficacy of sodium hypochlorite gel with twin kleen irrigating solution against enterococcus feacalis

The Enterococcus faecalis (E. faecalis) is a normal inhabitant of the oral cavity and is the most commonly identified species in post treatment asymptomatic persistent endodontic diseases (both intra-radicular and extra- radicular). Its prevalence ranges from 24% to 77%. To completely eliminate microorganisms from the root canal system, combination of instrumentation and irrigation with antibacterial solutions is necessary. Hence, the purpose of this study was to evaluate and compare the antibacterial efficacy of sodium hypochlorite gel with twin kleen irrigating solution against E. faecalis.40 non-carious, single-rooted teeth with straight canals were selected and instrumented till F3 (30.06) ProTaper Universal (Dentsply Maillefer, Ballaigues, Switzerland). E faecalis was prepared and incubated in teeth and were randomly divided into four groups of 10 each, Group A – 5.25% NaOCl gel, Group B – Twin Kleen Irrigating solution, Group C – 3% NaOCl solution and Group D – Saline. Sterile paper points were used to collect the samples after irrigation and then they were transferred to blood agar culture plates to check the presence of E. faecalis. Significant statistical difference was seen between the four groups. The effect of 5.25% NaOCl gel was superior as compared to Twin Kleen irrigating solution.Under the limitations of this study, 5.25% NaOCl gel was effective in reducing E. faecalis count followed by Twin kleen irrigating solutions.

First Report of Pantoea anthophila Causing Shoot Dieback Disease of Ceylon Ironwood (Mesua ferrea) in Malaysia

Ceylon ironwood (Mesua ferrea Linn.) or Penaga lilin is one of Asia's most popular tropical herbal plants, including Malaysia (Sharma et al., 2017). The trees are cultivated for their aesthetic value and pharmacological properties, especially as traditional remedies for asthma, dermatopathy, inflammation, and rheumatic conditions (Adib et al., 2019). In August 2022, a disease survey was conducted on Ceylon ironwood trees ranging from 5 to 12 years old in Botanical Park, Putrajaya, Malaysia, with 80% exhibiting shoot dieback disease of the 15 trees exhibiting shoot dieback disease. Symptoms include irregular, water-soaked with brown lesions on young leaves and shoots, where the small lesion coalesced and formed broad necrotic regions, subsequently causing dieback and gradual defoliation. Three infected shoots were collected from each tree, excised into small pieces (10 to 20 mm), immersed with 75% ethanol for 3 min, washed with 2% NaOCl solution for 1 min, and rinsed twice for 1 min in sterilized distilled water. A 10 µl aliquot of the sample suspension was streaked onto nutrient agar (NA) and incubated for 24 h to 48 h at 35 °C. A total of 15 isolates with similar morphology were obtained, and each isolate was re-streaked three times to obtain pure colonies that were round, smooth, with irregular edges, and produced yellow pigment in culture. All isolates were Gram-negative, negative for indole production, and utilized glucose, maltose, trehalose, sucrose, D-lactose, and pectin. Three representative isolates (C001, C002, and C003) with similar morphology were selected for further characterization. The total genomic DNA of all isolates was extracted from overnight cultures using Geneaid™ DNA Isolation Kit (Geneaid Biotech Ltd., Taiwan). PCR amplification of 16S rDNA (Zhou et al., 2015) and species-specific infB (Brady et al., 2008) genes was performed, and each of the ~1500 bp and ~900 bp amplicons were sequenced. BLASTn and phylogenetic analyses revealed all isolates were 100% identical to Pantoea anthophila (P. anthophila) LGM 2558 strains (Accession Nos. NR_116749 and NR_116113) for the 16S rDNA gene. They were 99% identical to P. anthophila CL1 strain (Accession Number CP110473) for infB gene. These sequences were later deposited in the GenBank (Accession Nos. OQ772233, OQ772234, and OQ772235 for 16S rDNA gene, and OQ803527, OQ803528, and OQ803529 for infB gene). For the pathogenicity test, healthy Ceylon ironwood seedlings' shoots were inoculated with 10 mL of each isolate suspension (1 x 108 CFU/ml) by spraying the inoculum on the young shoots using a sterilized spray bottle. Control seedlings were inoculated with sterile water. The inoculated shoots were covered with a sealed plastic bag to maintain the moisture and were kept in the greenhouse with temperatures ranging from 26 to 35 °C. The experiments were repeated twice, with three replicates for each treatment. Inoculated shoots showed dieback symptoms like natural infection, including irregular, water-soaked, and brown lesions on leaves and young shoots at 10 days post-inoculation. Control seedlings remained asymptomatic. The pathogen was re-isolated and identified via sequencing of the 16S rDNA and infB genes, thus fulfilling Koch’s postulates. Previously, P. anthophila has been reported to cause soft rot in wampee plants in China (Zhou et al., 2015) and leaf blight of cotton in Pakistan (Tufail et al., 2020). To our knowledge, this is the first report of P. anthophila causing shoot dieback disease of Ceylon ironwood trees in Malaysia. Plant disease management strategies need to be established to reduce losses due to P. anthophila infection since the pathogen could limit Ceylon ironwood tree production in Malaysia.