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Objective The persistence of pathogenic microorganisms in root canals is the most common reason for the failure of root canal treatment and the necessity of a root filling treatment, which results in an uncertain prognosis due to technical complexity and the variety of highly adaptable microorganisms. This study evaluated the effect of passive ultrasonic irrigation (PUI) on the outcome of the microbial analysis of root canal-treated teeth with persistent or recurrent apical inflammation in vivo. Materials and methods Sample collection was performed after root filling removal (sample S1, control group) and after PUI with NaCl (sample S2) using sterile paper points. In total, 19 samples were obtained. Quantification was performed by means of serial dilution of the samples. Subcultivated pure cultures were identified using MALDI-TOF MS complemented by the Vitek-2-System or PCR, followed by sequencing of the 16S rRNA gene. The results of the samples (S1 and S2) were evaluated regarding their bacterial count and composition. Results The total count of bacteria and the number of aerobic/facultative anaerobic microorganisms significantly increased in the S2-samples after application of PUI. The number of obligate anaerobic microorganisms showed an increase after PUI, although it was not significant. We detected 12 different aerobic/facultative anaerobic microorganisms before PUI, and in 21 cases after PUI. Two different obligate anaerobic microorganisms were found in S1 samples compared to nine different species in S2 samples. Conclusions PUI is a powerful method for detaching bacteria in infected root canals and enables a more precise analysis of the etiology of persistent endodontic infections. Clinical relevance This study indicates that PUI exerts a positive cleansing effect and adds to the accessibility of microorganisms during the application of bactericidal rinsing solution in root canal treatments.

Background New and improved surgical techniques are warranted to treat osteoarthritis of the thumb carpometacarpal joint (CMC-1). The Pyrocardan implant yields striking results but only few series exist, making the evidence scarce. Purpose The aim of this study was to conduct a prospective series using the Pyrocardan implant. Methods We compared the outcomes to a matched historical control group of patients operated on with ligament reconstruction and tendon interposition. The hypothesis was that the Pyrocardan implant would yield better patient-reported outcomes as well as the procedure would be safe and effective in relieving symptoms of CMC-1 osteoarthritis. In total, 30 patients were included in the prospective series. These 30 patients were compared, in a 1:3 design, to a matched historical group. Results Results were promising with visual analogue scale scores of 0.7 (rest) and 2.1 (function), key-pinch score of 5.1 kg, and shortened Disability of the Arm, Shoulder, and Hand score of 14.3 after 1 year when using the Pyrocardan implant. The revision rate was 10%. We found no evidence of subsidence of the thumb. We found no differences in patient-reported outcomes between the two groups. Conclusions In conclusion, the Pyrocardan implant is a viable option in the treatment of CMC-1 osteoarthritis but with a significant revision rate. When comparing the Pyrocardan implant to a historical control group, we failed to find any differences in patient-reported outcomes. Level of Evidence IV – case series.

Background: The role of diet and nutrition in the prevention of oral diseases has recently gained increasing attention. Understanding the influence of diet on oral microbiota is essential for developing meaningful prevention approaches to oral diseases, and the identification of typical and atypical responders may contribute to this. Methods: We used data from an experimental clinical study in which 11 participants were exposed to different dietary regimens in five consecutive phases. To analyse the influence of additional nutritional components, we examined changes in bacterial concentrations measured by culture techniques compared to a run-in phase. A measure of correspondence between the mean and individual patterns of the bacterial composition is introduced. Results: The distance measures introduced showed clear differences between the subjects. In our data, two typical and three atypical responders appear to have been identified. Conclusions: The proposed method is suitable to identify typical and atypical responders, even in small datasets. We recommend routinely performing such analyses.

With the rapid spread of SARS‐CoV2 has come a rapid proliferation of clinical research studies, resulting in considerable strain on research ethics committees (RECS), which need to review study proposals. RECs are pressured to move through the review process quickly so that studies can get underway to address the pandemic. These committees are also asked to increase efficiency without relaxing the standards for ethical review. RECs are accustomed to external pressure for approval from investigators; however, in the Covid‐19 era, this pressure is coming from not only the sponsors and investigators but also many other stakeholders, including world leaders, the community, the media, and professional organizations. Drawing on the authors' experiences on a central REC reviewing complex multicenter Covid‐19 studies, this commentary describes challenges that are inherent to Covid‐19 research studies, such as the difficulty of obtaining informed consent from patients ill with the highly infectious virus. The commentary recommends several steps that RECs can take to ensure ethical review of research studies during the Covid‐19 pandemic and future infectious disease outbreaks.

With the declaration of the COVID-19 pandemic by the World Health Organization in March 2020, many elements of society were faced with attempting to assimilate public health recommendations for infectious control. Vital social organizations had to balance delivering their social services while attempting to stay up to date with COVID-19 information and comply with evolving regulations. In the realm of schools and school systems, guidance on how to best adapt to COVID-19 was often limited. School officials and staff had to assist with multiple public health crises as a consequence of the pandemic, from the pandemic’s transmission prevention strategies (e.g., face masks and physical distancing) to the recognition that students would have personal tragedies related to COVID-19. In this review, we highlight the process and feasibility of implementing an international COVID-19 school-based initiative over two years of the pandemic, the Health Education and Training (HEAT) Corps program.

Introduction Surgical treatment of thumb trapeziometacarpal osteoarthritis usually involves 4 to 8 weeks of postoperative casting and splinting followed by varying mobilization protocols. Suspension arthroplasty has been described as an alternative to allow earlier range of motion exercises. The purpose of this study was to compare patient-reported outcomes (PRO) when adding a two-string suture-button suspension arthroplasty (Mini TightRope, MTR) to our usual procedure of ligament reconstruction and tendon interposition (LRTI), allowing early mobilization. Can we allow early mobilization using this technique without jeopardizing the PRO results at the 1 year follow-up and without an increased risk of complication? Materials and Methods A prospective study using the MTR system (Arthrex) as a suture-button suspensionplasty was conducted. Twelve patients (MTR group) and 36 historical patients (LRTI alone) were included. Results At 12 months, the median value for quick disabilities of the arm, shoulder, and hand was 11.3 (range, 0–43.2) in the MTR group and 13.6 (range, 0–88.6) in the LRTI group, resulting in similar improvements, p = 0.46. One in twelve patients in the MTR group was dissatisfied and 9 in 36 in the LRTI group were dissatisfied, p = 0.41. No complications were observed during the first year. Conclusion Supplemental suture-button suspensionplasty can be utilized for high demand patients and patients who want to reduce immobilization time without major complications and with similar PRO as LRTI at 6 and 12 months. Level of evidence Four case series

Pediatric neuropathic pain is caused by a spectrum of disorders that are generally challenging to treat. Many of the underlying altered neurological processes are being elucidated through mechanistic studies. Few randomized control trials have evaluated the use of opioids for the treatment of adult neuropathic pain conditions, and there have been none in pediatric populations. With sparse data to provide guidance and an incomplete understanding of the underlying mechanisms, the use of opioids remains unclear. Our clinical experience and typical risk versus benefit considerations suggest a limited, if any, role for using opioids to treat pediatric neuropathic pain. In this literature review, we review the available adult and pediatric data and provide general guidance on this subject matter.

AIM Aim of this study was to investigate the influence of an anti-inflammatory diet on different parameters in patients with gingivitis. MATERIALS AND METHODS Thirty patients were randomly allocated to an experimental and a control group stratified by their plaque values. The experimental group had to change to a diet low in processed carbohydrates and animal proteins, and rich in omega-3 fatty acids, vitamin C, vitamin D, antioxidants, plant nitrates and fibres for 4 weeks. The control group did not change their diet. Both groups suspended interdental cleaning. Periodontal parameters were assessed by a blinded dentist. Serological and subgingival plaque samples were taken at baseline and end. RESULTS While there were no differences regarding the plaque values, the experimental group showed a significant reduction in gingival bleeding (GI Baseline: 1.04 ± 0.21, GI End: 0.61 ± 0.29, p

Background Osteoarthritis of the thumb carpometacarpal joint (CMC-1 OA) is increasingly common with age. Conservative treatment with anti-inflammatory medication, thumb spica splinting, and steroid injection is recommended for early-stage OA, but some patients will continue to have refractory symptoms and surgery may be considered. We found it interesting to study outcomes of surgical treatment of cases with mild radiographic changes and yet symptoms severe enough to indicate surgery. The specific research question is, if there is a limit of radiographic changes, below which a poor patient-reported outcome (PRO) can be expected. Purpose In a retrospective cohort of patients with prospectively collected PROs, we intend to study the effect of the radiographic CMC-1 OA severity on the PRO improvement and satisfaction after interposition arthroplasty. Patients and Methods Radiographs of 347 patients, who had CMC-1 surgery, were retrospectively analyzed. Each rater independently assessed all radiographs classifying each according to the Eaton classification. All patients had surgery with CMC-1 interposition arthroplasty using three well-known techniques that all include a complete trapeziectomy. Comparison between stage 1 and stage 2–4 was done using Chi-square test and t-test. Results Patients with Eaton stage 1 had a mean improvement in Quick-DASH (Quick-Disability of the Arm, Shoulder, and Hand questionnaire) scores of 14.6 points. Patients with Eaton stage 2 to 4 had a mean improvement of 25.3 points. The difference between these two groups was 10.6 points (p = 0.009). Only 52% of patients with Eaton stage 1 OA were satisfied. However, 76% of patients with Eaton stage 2 to 4 were satisfied (p = 0.008, chi-square between stage 1 and stage 2–4). Conclusion Patients with Eaton stage 1 CMC-1 OA had poorer PROs, as compared with more advanced stages of OA, 6 months following surgical treatment with interposition arthroplasty. Based on our results, we advise against surgical treatment with interposition arthroplasty of the very mildest CMC-1 OA, regardless of the preoperative PROs.

Introduction To date, the microbiota associated with persistent endodontic infections has only been analyzed in patients who did not receive any antibiotic therapy for at least 3 months before endodontic treatment. In this clinical study, secondary endodontic infections of patients who recently received systematic antibiotic therapy before endodontic treatment were analyzed and compared with the actual data available in the literature. Methods Root canal-filled teeth with periradicular lesions of 20 patients who were under systematic antibiotic therapy which ended 1-21 days before the endodontic treatment were studied. A wide range of antibiotics was administered, including amoxicillin, amoclav, amoxicillin/metronidazole, ampiclox (ampicillin and cloxacillin), doxycycline, tetracycline, ciprofloxacin, and azithromycin. Microorganisms were isolated according to standard protocols and identified using MALDI-TOF-MS. A narrative review of the literature was conducted to compare the results of this study with the data reported so far. Results The presence and concentrations of bacteria isolated from the infected root canals were comparable with those depicted in the literature, although the total colony-forming units number in saliva was rather low. The number of different bacterial species isolated and identified in each patient as well as the diversity over all patients did not show signs of any influence of the administered antibiotics. Weissella hellenica and Cellulomonas spp. were detected in root canals for the first time. Granulicatella adiacens and Dietzia spp., previously isolated from primary endodontic infections, were detected for the first time in persistent root canal infections in this patient group. Conclusions The bacterial diversity reported to date in secondary endodontic infections should be extended with the new microbial composition revealed in endodontic patients who had recently received systematic antibiotic therapy.

Caries development is associated with shifts in the oral biofilm microbiota and primarily linked to frequent simple carbohydrate consumption. Different nutritional ingredients can either promote or prevent caries development. To investigate the effects of selected ingredients on the oral biofilm microbiota in situ, 11 study participants underwent 3-month-long dietary phases with intake of a regular diet (PI), additional frequent sucrose (PII), milk and yoghurt (PIII), and a diet rich in dietary fiber (PIV) and then returned to their regular diet (PV). Oral biofilm was sampled and analyzed applying 16S rRNA Illumina MiSeq sequencing. Additionally, the effect on the enamel was analyzed by measuring enamel surface roughness with laser scanning microscopy. The beta-diversity results showed that the microbiota in all the following phases differed significantly from PI and that the microbial community in PII was significantly different from all other phases. The abundance of the genus Streptococcus fluctuated over the course of the five phases, with a significant increase in PII (P = 0.01), decreasing in PIII and PIV (PIII and PIV versus PII: P < 0.00001) and increasing again toward PV. Other taxa showed various fluctuations of their abundances, with PV returning approximately to the levels of PI. In conclusion, while elevated sucrose consumption favored caries-promoting non-mutans streptococci, frequent milk and yoghurt intake caused a significant decrease in the abundance of these microbial taxa and in addition reduced enamel surface roughness. These results indicate that modulations of the oral biofilm microbiota can be attained even in adults through dietary changes and corresponding recommendations can be made for the prevention of caries development.IMPORTANCE Caries affects a large proportion of the population worldwide, resulting in high treatment costs. Its etiology can be ascribed to shifts of the microbiota in dental biofilms primarily driven by dietary factors. It is unclear how diet affects the microbial community of plaque biofilm in situ and whether it can be modulated to help prevent caries development. To address these issues, we analyzed changes of the in situ plaque microbiota following 3-month-long dietary changes involving elevated sucrose, dairy, and dietary fiber consumption over a period of 15 months. Applying high-throughput sequencing, we found non-mutans streptococci, a taxonomic group involved in the beginning stages toward microbial dysbiosis, in decreased abundance with elevated dairy and dietary fiber intake. Through analysis of the enamel surface roughness, these effects were confirmed. Therefore, correspondent dietary measures can be recommended for children as well as adults for caries prevention.

The influence of a change in nutrition on the oral microbiota are discussed in literature, but usually only changes of population mean values are reported. This paper introduces simple methods to also analyse and report the variability of patients&rsquo, reactions considering data from the culture analysis of oral biofilm. The framework was illustrated by an experimental study exposing eleven participants to different nutrition schemes in five consecutive phases. Substantial inter-individual variations in the individual reactions were observed. A new coherence index made it possible to identify 14 instances where the direction of individual changes tended to coincide with the direction of the mean change with more than 95% probability. The heterogeneity in variability across different bacteria species was limited. This allowed us to develop recommendations for sample sizes in future studies. For studies measuring the concentration change of bacteria as a reaction to nutrition change, the use of replications and analysis of the variability is recommended. In order to detect moderate effects of a change in nutrition on the concentration of single bacterial taxa, 30 participants with three repetitions are often adequate. Insights into the relationship between nutrition and the microbial composition can be helpful for the development of dietary habits that promote the establishment of a healthy microbial flora and can therefore prevent the initiation of oral diseases such as caries and periodontitis.

To date, the microbiota associated with persistent endodontic infections has only been analyzed in patients who did not receive any antibiotic therapy for at least 3 months before endodontic treatment. In this clinical study, secondary endodontic infections of patients who recently received systematic antibiotic therapy before endodontic treatment were analyzed and compared with the actual data available in the literature.Root canal-filled teeth with periradicular lesions of 20 patients who were under systematic antibiotic therapy which ended 1-21 days before the endodontic treatment were studied. A wide range of antibiotics was administered, including amoxicillin, amoclav, amoxicillin/metronidazole, ampiclox (ampicillin and cloxacillin), doxycycline, tetracycline, ciprofloxacin, and azithromycin. Microorganisms were isolated according to standard protocols and identified using MALDI-TOF-MS. A narrative review of the literature was conducted to compare the results of this study with the data reported so far.The presence and concentrations of bacteria isolated from the infected root canals were comparable with those depicted in the literature, although the total colony-forming units number in saliva was rather low. The number of different bacterial species isolated and identified in each patient as well as the diversity over all patients did not show signs of any influence of the administered antibiotics. Weissella hellenica and Cellulomonas spp. were detected in root canals for the first time. Granulicatella adiacens and Dietzia spp., previously isolated from primary endodontic infections, were detected for the first time in persistent root canal infections in this patient group.The bacterial diversity reported to date in secondary endodontic infections should be extended with the new microbial composition revealed in endodontic patients who had recently received systematic antibiotic therapy.

While appreciation of the fundamental role biodiversity plays in underpinning the long-term sustainability of agricultural systems is growing, international commitments to preserve and protect this natural resource have not yet been achieved. Loss and degradation of farmland habitats are an important contributor to the continued decline of biodiversity. Despite this, little information is available regarding the diversity and ecological condition of farmland habitats in Europe. Indeed, where habitat data are available, this is usually at a very broad landscape scale rather than farm scale. Coupled with this, strategies to increase agricultural output in response to growing global population, will likely place increasing pressure on farmland biodiversity. Knowledge and ongoing monitoring of farmland habitat type and extent is a prerequisite for the future protection of much of Europe’s biodiversity. Here we report the findings of a national scale survey of farmland habitat diversity in Ireland. Detailed surveys of farm habitats and management practices (system, stocking rate agri-environment scheme participation status and organic N and P inputs) were undertaken on 118 farms in three regions of the Republic of Ireland (RoI). Recorded farm habitats were subsequently digitised on orthophotography. The resulting ground truthed data from the total surveyed area of 3688 ha, were then used, together with satellite imagery, to classify the habitat composition of a further approximately 87,000 ha of the surrounding landscape. Results revealed that at individual farm scale, an average of 73% of the land surveyed comprised agriculturally productive (mainly improved grassland) habitats. Marginally productive habitats (mainly extensively managed grasslands) accounted for an average of 11% of farm area, while other semi-natural habitats (mainly hedgerows) accounted for an average of 13%, with the remaining ca. 3% under build ground. Results from the classification at the wider landscape scale showed a similarly substantial incidence of non-intensively managed habitats. However, at both farm and landscape scales, habitat diversity was found to vary markedly between different regions and farming systems. This study represents one of a very small number that currently exist, where farm scale habitat and management data have been collected. From the few that are available, semi-natural habitat cover has been found to account for an average farm area of 1–12%. Therefore, our data present a relatively positive picture in terms of the intensity, scale and impact of Irish farming on landscape heterogeneity. However, as in other parts of the world, agricultural expansion and intensification to meet increased global food supply will necessitate careful monitoring of the impact of these changes on the structure of farmed landscapes. This study provides a novel approach for the collection of such monitoring data at farm scale, and illustrates how such data can be reliably up-scaled to landscape level.

Background Oral malodor is a very discomforting condition deriving from the presence of volatile sulfur compounds in the expired air. In halitosis of intraoral etiology, the volatile sulfur compounds are metabolic products of the oral microorganisms within the biofilm coating the tongue dorsum as well as other tissues in the oral cavity. The aim of this study was to characterize and compare the microbial composition of tongue biofilm in volunteers suffering from halitosis and healthy volunteers by means of both the culture method and culture‐independent cloning technique. Results A high bacterial variety (more than 80 different species) was detected using the combination of both methods. A distinct bacterial composition was revealed in the halitosis‐associated biofilms compared with the health‐associated biofilms. Actinomyces graevenitzii was shown to be significantly associated with the halitosis condition. The culture method identified 47 species, included Veillonella rogosae, never isolated from the tongue biofilm of halitosis patients so far. In the healthy condition, the culture‐dependent method showed that the most frequent species were Streptococcus parasanguinis among the aerobes and Veillonella spp. among the anaerobes. The culture‐independent cloning method detected more than 50 species. Streptococci, in particular S. mitis/oralis, S. pseudopneumoniae, and S. infantis as well as Prevotella spp., were found most frequently in halitosis patients. Streptococcus salivarius and Rothia mucilaginosa were found more frequently in the healthy condition. Conclusions The combination of the culture‐dependent and culture‐independent cloning techniques allowed for a widespread analysis of the tongue biofilm in halitosis patients. The results can support further pharmacological research for new antimicrobial agents and halitosis therapy strategies., Oral malodor is a discomforting condition deriving from the presence of volatile sulfur compounds in the expired air. The study characterized and compared the microbial composition of tongue biofilm in volunteers suffering from halitosis and healthy volunteers by means of both the culture method and culture‐independent cloning technique, revealing a distinct bacterial composition of halitosis‐associated biofilms compared with the health‐associated biofilms and a high bacterial diversity within the halitosis‐associated biofilms.

Background: Investigating the influence of nutrition on oral health has a long scientific history. Due to recent technical advances like sequencing techniques for the oral microbiota, this topic has gained scientific interest again. A basic challenge is to understand the influence of nutrition on the oral microbiota and on the interaction between the oral bacteria, which is also statistically challenging. Methods: Log-transformed ratios of two bacteria concentrations are introduced as the basic analytic tool. The framework is illustrated by application in an experimental study exposing eleven participants to different nutrition schemes in five consecutive phases. Results: The method could be sufficiently used to analyse the interrelation between the bacteria and to identify some bacterial groups with the same as well as different reactions to additional dietary components. It was found that the strongest changes in bacterial concentrations were achieved by the additional consumption of dairy products. Conclusion: A log ratio-based analysis offers insights into the relation of different bacteria while taking specific features of compositional data into account. The presented methods allow becoming independent of the behaviour of other bacteria, which is a disadvantage of common analysis methods of compositions. The results indicate that modulations of the oral biofilm microbiota due to nutrition change can be attained.

Purpose. Micro-organisms are important triggers of peri-implant inflammation and analysing their diversity is necessary for peri-implantitis treatment. This study aimed to analyse and compare the microbiota associated with individuals with peri-implantitis, as well as clinically healthy implant sites. Methodology. Subgingival biofilm samples were taken from 10 individuals with peri-implantitis and from at least 1 clinically healthy implant. DNA was extracted and bacterial 16S rRNA genes were amplified using universal primers. After cloning the PCR-products, amplified inserts of positive clones were digested using restriction endonucleases, and the chosen clones were sequenced. The 16S rDNA-sequences were compared to those from the public sequence databases GenBank, EMBL and DDBJ to determine the corresponding taxa. Results. Differing distributions of taxa belonging to the phyla Firmicutes, Bacteroidetes, Fusobacteria, Actinobacteria, Proteobacteria, Synergistetes, Spirochaetae and TM 7 were detected in both the healthy implant (HI) and the peri-implantitis (PI) groups. A significantly higher relative abundance of phylum Bacteroidetes, as well as of the species Fusobacterium nucleatum, were found in the PI group (P

Enterococcus faecalis, a commensal of the intestinal tract of humans and animals is of great significance as leading opportunistic pathogen, and also prevalent in oral diseases, such as endodontic infections, as well as the healthy oral cavity. To investigate the potential of oral E. faecalis to constitute a reservoir of antibiotic resistance, isolates from supragingival plaque/saliva and from endodontic infections were screened regarding their resistance to selected antibiotics in comparison to nosocomial and food isolates.

The green peach aphid, Myzus persicae (Sulzer), is a serious pest throughout the world, attacking a broad range of crop plants across numerous agricultural industries. This species has a high propensity to develop chemical resistance, and has the unenviable title of having resistance to more insecticides than any other insect species. An extensive survey of field populations was undertaken across Australia, and showed widespread and high levels of resistance to carbamates and synthetic Pyrethroids in M. persicae. Moderate levels of resistance to organophosphates were also observed in many populations, while there is new evidence of resistance developing to neonicotinoids. Isofemale (clonal) lines of M. persicae were generated and subsequently tested across a range of insecticides; individual genetic clones were found to contain resistance to multiple chemical classes. Resistance genotyping of these aphids were consistent with published literature of known resistant mechanisms. The high and widespread levels of resistance identified within Australia are concerning. Resistance in M. persicae has spread quickly across Australia, and thus farmers are likely to have fewer chemical control options in the future. There is a need to develop resistance management strategies that rotate insecticides, spray insecticides only when economically necessary, and incorporate nonchemical control options.

Introduction To date, a variety of microbial species have been isolated from endodontic infections. However, endodontic clinical bacterial isolates have not been sufficiently characterized with regard to their capacity for antibiotic resistance and biofilm formation. In this study, antibiotic resistance and biofilm formation of 47 different aerobic and anaerobic bacterial isolates, belonging to 32 different species previously isolated from infected filled root canals, were studied. Methods Antibiotic sensitivity to 11 antibiotics including penicillin G, amoxicillin, clindamycin, gentamicin, vancomycin, tetracycline, doxycycline, fosfomycin, rifampicin, ciprofloxacin, and moxifloxacin was tested using the standardized Etest method (Bio Merieux, Marcy-1'Etoile, France). The antibiotic sensitivity of 4 control strains was also estimated in parallel. Additionally, the capacity to form biofilms was quantified using the microtiter plate test. Results Different aerobic and anaerobic bacterial species were either resistant against a number of antibiotics or showed high minimal inhibitory concentrations against clinically relevant antibiotics. Five aerobic and 2 anaerobic isolates, including Enterococcus faecalis , Streptococcus mutans , Lactobacillus fermentum , Actinomyces naeslundii , Actinomyces viscosus , Prevotella buccae , and Propionibacterium acidifaciens , were characterized as being high biofilm producers, whereas 8 aerobic and 3 anaerobic isolates were found to be moderate biofilm producers. Most isolates with resistance or markedly high minimal inhibitory concentration values were also either moderate biofilm producers or high biofilm producers. Conclusions These results suggest that the clinical significance of endodontic infections could include that they serve as a reservoir for antibiotic resistance. Furthermore, endodontic treatment should consider the adhesion and biofilm formation by a variety of bacteria.

Field margins within intensively managed ecosystems are often seen as a last refuge for biodiversity, and are typically targeted with measures within many agri-environment schemes. Grassland accounts for 81 million ha of land within the EU; however, the ecology of field margins associated with permanent grassland has not been well studied. This study investigated the effects of experimental field margin measures on hymenopteran parasitoid communities over a 5-year period. Hymenopteran parasitoids were chosen because they occupy high trophic levels, feed on a diverse range of plant and invertebrate hosts, and are considered good indicators of arthropod diversity. Establishment methods included: fencing, natural regeneration, and reseeding with a wildflower mixture, at three margin widths. Field margin establishment method had a significant effect on abundance of parasitoids. Establishment method did not have a significant effect on parasitoid genera diversity, but did affect parasitoid community composition. Margin width had no significant impact on parasitoid communities. Grazing had a significant negative effect on parasitoid genus richness and community structure. This suggests that structural diversity of vegetation plays an important role in parasitoid community structure. Plant species richness did not significantly affect parasitoid abundance or genera richness. Noxious species within plots resulted in a significantly greater abundance and diversity of parasitoids and of idiobionts in particular. Where plant species richness is limited, simple measures such as fencing of narrow field margins may be as effective at increasing parasitoid taxon richness and abundance (indicators of arthropod richness and abundance) as expensive measures such as reseeding and wider margin widths.

Maternity claims, as a group, are the most expensive clinical negligence claims reported to the NHS Litigation Authority (NHSLA) and the second highest by volume. As a consequence, the NHS LA launched a project Ten Years of Maternity Claims – An Analysis of NHS Litigation Authority Data reviewing maternity claims with an incident date between 1 April 2000 and 31 March 2010. This analysis was initiated to enable key trends to be identified, and to share the learning widely within the National Health Service (NHS) with a long-term goal of reducing harm occurring to women and babies accessing maternity care. During this period there were 5087 claims with a total value of £3.1 billion. Between the years 2000 and 2009 there were 5.5 million births in England. In all, 0.1% of births were subject to a clinical negligence claim. In other words, the vast majority of births do not result in a clinical negligence claim, which supports the view that having a baby within the NHS is very safe. As part of the project, primary and secondary level studies were undertaken. The former analysed the occurrence of types of maternity claim and the financial costs associated with them. The most frequent categories identified related to the management of labour, which involves fetal heart rate monitoring (14.05%), Caesarean section (13.24%) and cerebral palsy (10.65%). The secondary level study reviewed four categories of maternity claims which were chosen subjectively from discussions with stakeholders. The categories identified were – antenatal ultrasound investigations, cardiotocograph interpretation, perineal trauma and uterine rupture. By undertaking additional in depth analysis the objective was to identify factors from which risk management recommendations could be formulated to facilitate learning from claims, with the aim of reducing cases of actual harm to mothers and babies. This article highlights the key findings from the study and suggests recommendations for ongoing learning within maternity services.

1. Arthropods, a major component of functional biodiversity within agro-ecosystems, contribute to sustainability through processes including nutrient cycling and pest control. Extensively managed field margins can help protect this functional biodiversity by providing habitat for beneficial species. 2. This 2 year study investigated the relative benefits of grassland field margin treatments (Fenced only, Rotavated, and Reseeded (with a grass and wildflower mixture)) on the abundance and taxon richness of five arthropod trophic groups (detritivores, herbivores, predators, parasitoids and hyperparasitoids) from the orders Coleoptera, Hemiptera, Hymenoptera, Diptera and Araneae. 3. The taxon richness and abundance of all trophic groups (with the exception of herbivore abundance) was greater in fenced field margin treatments than in the adjacent grazed field, particularly by the final sampling occasion. However, there were seasonal differences, with abundance and taxon richness generally greater in August than in June. Only detritivores and herbivores responded to the individual fenced field margin treatments. The botanically more species rich, rotavated and reseeded treatments, had greater detritivore and herbivore richness and abundance than the fenced only treatments. 4. Community structure analysis indicated that the grazed (within) field and fenced field margins had two distinct communities for all trophic groups, demonstrating the conservation value of the margins within intensively managed agricultural grasslands. 5. The current Irish agri-environment scheme requires fencing of field margins. Our results highlight how this relatively simple measure can benefit arthropod conservation and functional biodiversity. This may in turn benefit farm productivity through potentially improved nutrient cycling and natural pest control.

In this study, habitat surveys were undertaken on 50 grass-based farms in SE Ireland and data digitised onto aerial photography. Additional data i.e. stocking rates, and participation (or otherwise) in the Irish Rural Environment Protection Scheme (REPS) were collected and analysed as possible explanatory variables for farm habitat composition. Results indicated that approximately 14.3% of the land area of sampled farms comprised of semi-natural habitat types, a proportion substantially greater than has been reported for many other European countries. The most frequently recorded semi-natural habitats included, field boundaries, scrub, and deciduous and riparian woodlands. Multivariate analysis of farm habitat configuration showed a strong dichotomy between dairy and non-dairy farming systems. Habitats such as intensively managed grassland and built ground were closely associated with dairy-based enterprises. In contrast, the incidence of other habitat types was associated with non-dairy and/or REPS participating enterprises. The ecological quality of field boundaries as assessed by the Field Boundary Evaluation and Grading System (FBEGS) Index was significantly greater on dairy, compared with dry-stock farms. This dichotomy in farm habitat composition is not reflected within current Agri-Environment (AE) policy. Integration of locally important drivers of habitat diversity into the design and implementation of AE policy, is integral to the successful performance of AE schemes.

Summary 1. As measuring biodiversity in its entirety is impractical, there is a need for bioindicators. This study tested the hypothesis that parasitoid Hymenoptera are potential bioindicators that provide a useful means to assess the wider biodiversity of arthropod populations in agro-ecosystems. There are a range of theoretical arguments to support such a claim, including the high trophic position of these taxa within the arthropod communities in which they occur, and the unique nature of their biological relationships with the majority of terrestrial arthropod groups. 2. A survey of 48 commercial farms was conducted and Generalized Linear Models used to investigate relationships between six taxa—parasitoid Hymenoptera, Coleoptera, Hemiptera, Diptera, Araneae and plants (species richness and sward height)—in agricultural grasslands. As well as relationships between these groups, the relationship of each individual group to the overall biodiversity of all other arthropod groups was explored. 3. Both abundance (r 2 =0 AE58) and taxon richness (r 2 =0 AE54) of parasitoid Hymenoptera had stronger relationships with overall arthropod taxon richness than any other arthropod group investigated. Parasitoid abundance also had a positive relationship with species richness of Coleoptera (r 2 =0 AE23) and Hemiptera (r 2 =0 AE47). 4. An historical data set demonstrated how the relationship between parasitoid abundance and overall arthropod taxon richness changes over the growing season. July, when the relationship was strongest, is potentially the most useful time to sample. 5. For use in routine monitoring, it is important that an effort be made to understand the seasonal influence on the relationship in the context being studied. Equal sampling effort must be made for all sites being compared and sites should be sampled as close together in the season as is possible. 6. Synthesis and applications We show that, within agricultural grasslands, both the abundance and taxon richness of parasitoid Hymenoptera are more closely related with overall arthropod diversity than any other arthropod group investigated. The use of parasitoid abundance provides a simple and practicable monitoring tool for tracking change in wider arthropod diversity in agro-ecosys

1. It is well documented that cattle reduce their grazing activity in the vicinity of cattle dung, which gives rise to distinct patches, or islets as they have been termed, of longer sward. The influence of such islets on pasture utilisation and agronomic performance has been widely studied, but very little information is available concerning their influence on grassland biodiversity. 2. In this study the abundance and distribution of arthropods in relation to islets was assessed, using suction sampling, at 26 commercial farms and in a replicated pasture management experiment in the south and east of Ireland. 3. Islets were found to cover approximately 24% of pastures and to contain between 40% and 50% of arthropod individuals. 4. Islets consistently contained a higher density of arthropods, even when the difference in mean sward height between islets and more strongly grazed sward was accounted for. The relative concentration of arthropods in islets declined with increasing mean sward height, which may be related to the recovery of well-grazed non-islet sward. Islets appear to act as refugia from sward removal. 5. The potential importance of islets in maintaining arthropod biodiversity within intensively grazed pastures and the wider grass-based farming landscape is discussed, particularly with reference to standard agronomic practices such as sward topping and chain harrowing, which aim to remove the sward heterogeneity created by grazing livestock.

A 5′ nuclease duplex real-time polymerase chain reaction (PCR) assay was developed and validated with various food products for the specific and fast detection of Salmonella spp. in food. The assay used previously published primers in combination with a newly developed probe targeting the invA gene. An internal amplification control, which is coamplified in a duplex PCR, was included in the assay. The analysis of 1,934 natural food samples with real-time PCR and the cultural method in parallel resulted in a relative accuracy of 100% and 99.84% respectively, depending on the enrichment procedure in which buffered peptone water and selective enrichment in Rappaport–Vassiliadis (RV) broth were employed. The duplex real-time PCR assay has proven to be a specific, sensitive and fast screening method for Salmonella spp. in food. The overall analysis time of the PCR method was approximately 28 h, in contrast to 4 to 5 days with conventional Salmonella diagnostics. The developed assay has been shown to be a reliable diagnostic tool for use in routine analysis. It has been validated thoroughly and has become an official method in Germany for the detection of Salmonella spp. in food.

In this work the intra and inter-laboratory validation of a duplex real-time PCR screening method for the detection of genetically modified (gm) plants is described. Target DNA sequences from Cauliflower Mosaic Virus 35S promoter (P35S) and nos-terminator from Agrobacterium tumefaciens (T-nos) are amplified. The duplex real-time PCR method is using primer and probe sequences that have already been published for the individual (“single”) detection of both target sequences. The validation showed sensitivity comparable to the single PCR standard methods. In addition, combined with a reference gene and using reference standard material, the method can be used to semiquantitatively estimate the amount of gm plants in an unknown sample.

A collaborative study including 13 German laboratories was conducted to evaluate the performance of two non-patented real-time PCR methods for the detection of Salmonella in milk powder targeting the ttrC/ttrA- or the invA gene. The enrichment procedure and sample DNA preparation method prior to the real-time PCR was the same for both systems and the identical DNA extraction samples were analysed. The traditional cultural method according to EN ISO 6579:2002 for the detection of Salmonella in food was performed in each laboratory as the reference. The participants received twelve coded milk powder samples each of 25 g for the analysis. Four of them were Salmonella negative (level L0), four artificially contaminated with

peer-reviewed We compared arthropod taxon richness, diversity and community structure of two replicated grassland husbandry experiments to investigate effects of reduced management intensity, as measured by nutrient input levels (390, 224 and 0 kg/ha per year N in one experiment, and 225 and 88 kg/ha per year N in another). Suction sampling was used to collect Araneae, Coleoptera, Hemiptera and Hymenoptera, with Araneae and Coleoptera also sampled with pitfall trapping. Univariate analyses found no significant differences in abundance and species density between treatments. However, with multivariate analysis, there were significant differences in arthropod community structure between treatments in both experiments. Reducing N input and associated stocking rates, as targeted by agri-environment schemes, can significantly alter arthropod communities but without increasing the number of species present. Other approaches that may be necessary to achieve substantial enhancement of sward arthropod biodiversity are suggested. This work was part of the Ag-Biota Project, funded by the Environmental Protection Agency, Ireland (2001-CD ⁄ B1-M1) through the ERTDI Programme under the National Development Plan (2000–2006).

Antimicrobial photodynamic therapy (APDT) has gained increased attention as an alternative treatment approach in various medical fields. However, the effect of APDT using visible light plus water-filtered infrared A (VIS + wIRA) on oral biofilms remains unexplored. For this purpose, initial and mature oral biofilms were obtained in situ ; six healthy subjects wore individual upper jaw acrylic devices with bovine enamel slabs attached to their proximal sites for 2 h or 3 days. The biofilms were incubated with 100 μg ml −1 toluidine blue O (TB) or chlorin e6 (Ce6) and irradiated with VIS + wIRA with an energy density of 200 mW cm −2 for 5 min. After cultivation, the CFU of half of the treated biofilm samples were quantified, whereas following live/dead staining, the other half of the samples were monitored by confocal laser scanning microscopy (CLSM). TB- and Ce6-mediated APDT yielded a significant decrease of up to 3.8 and 5.7 log 10 CFU for initial and mature oral biofilms, respectively. Quantification of the stained photoinactivated microorganisms confirmed these results. Overall, CLSM revealed the diffusion of the tested photosensitizers into the deepest biofilm layers after exposure to APDT. In particular, Ce6-aided APDT presented elevated permeability and higher effectiveness in eradicating 89.62% of biofilm bacteria compared to TB-aided APDT (82.25%) after 3 days. In conclusion, antimicrobial photoinactivation using VIS + wIRA proved highly potent in eradicating oral biofilms. Since APDT excludes the development of microbial resistance, it could supplement the pharmaceutical treatment of periodontitis or peri-implantitis.

The green peach aphid, Myzus persicae (Sulzer), is a serious pest throughout the world, attacking a broad range of crop plants across numerous agricultural industries. This species has a high propensity to develop chemical resistance, and has the unenviable title of having resistance to more insecticides than any other insect species. An extensive survey of field populations was undertaken across Australia, and showed widespread and high levels of resistance to carbamates and synthetic pyrethroids in M. persicae. Moderate levels of resistance to organophosphates were also observed in many populations, while there is new evidence of resistance developing to neonicotinoids. Isofemale (clonal) lines of M. persicae were generated and subsequently tested across a range of insecticides; individual genetic clones were found to contain resistance to multiple chemical classes. Resistance genotyping of these aphids were consistent with published literature of known resistant mechanisms. The high and widespread levels of resistance identified within Australia are concerning. Resistance in M. persicae has spread quickly across Australia, and thus farmers are likely to have fewer chemical control options in the future. There is a need to develop resistance management strategies that rotate insecticides, spray insecticides only when economically necessary, and incorporate nonchemical control options.

Root canal treatment failures often correlate with persistent biomaterial-associated endodontic infections. The aim of the present study was to assess the impact of endodontic obturation material sampling from root canals with posttreatment apical periodontitis on improving standard study protocols. Samples from previously filled root canals and their corresponding endodontic filling materials were obtained from five root-filled teeth with posttreatment periradicular lesions. After cultivation, the isolated microorganisms were quantified and biochemically identified. Moreover, clone libraries were constructed after the amplification of bacterial 16S ribosomal DNA (rDNA) from the same samples. DNA from selected clones was sequenced to identify microbial species. Transmission electron microscopy (TEM) aided visualization of the detected bacteria. Overall, 22 taxa of the phyla Firmicutes, Actinobacteria, and Bacteroidetes were detected in both obturation and root canal samples by culture-dependent and culture-independent methods. Root canal fillings sheltered 17 species (3.30–7.50 × 103 CFU/ml). Of these, nine were detected solely in the retrieved obturation materials. The reinfected root canals harbored 13 taxa (3.48–7.36 × 103 CFU/ml). Obligate and facultative anaerobic bacteria prevailed. The number of different species ranged from 1 to 5 within a single sample. Fungi were not detected. Bacteria can colonize both root canals and endodontic fillings in vivo. Integrating the sampling of obturation materials with standard root canal sample collection offers a clearer insight into the actual microbial flora of reinfected root canals and improves the study protocols of secondary/persistent endodontic infections.

A microscopic method for noninvasively monitoring oral biofilms at the macroscale was developed to describe the spatial distribution of biofilms of different bacterial composition on bovine enamel surfaces (BES). For this purpose, oral biofilm was grown in situ on BES that were fixed at approximal sites of individual upper jaw acrylic devices worn by a volunteer for 3 or 5 days. Eubacteria, Streptococcus spp., and Fusobacterium nucleatum were stained using specific fluorescence in situ hybridization (FISH) probes. The resulting fluorescence signals were subsequently tested by confocal laser scanning microscopy (CLSM) and monitored by an automated wide-field microscope-based imaging platform (Scan∧R). Automated image processing and data analysis were conducted by microscope-associated software and followed by statistical evaluation of the results. The full segmentation of biofilm images revealed a random distribution of bacteria across the entire area of the enamel surfaces examined. Significant differences in the composition of the microflora were recorded across individual as well as between different enamel surfaces varying from sparsely colonized (47.26%) after 3 days to almost full surface coverage (84.45%) after 5 days. The enamel plates that were positioned at the back or in the middle of the oral cavity were found to be more suitable for the examination of biofilms up to 3 days old. In conclusion, automated microscopy combined with the use of FISH can enable the efficient visualization and meaningful quantification of bacterial composition over the entire sample surface. Due to the possibility of automation, Scan∧R overcomes the technical limitations of conventional CLSM.

Ecologically, potential bioindicator taxa operate at different scales within agricultural ecosystems, and thereby provide a means to investigate the influence of changing management practice on biological diversity at different scales within the agro-ecosystem. Surveys of grassland plant species at field level, parasitoid Hymenoptera at the field and farm scale, and bird populations and habitats at farm scale were carried out on 119 grass-based farms across three regions in the Republic of Ireland. In addition, habitat richness and aquatic macroinvertebrates were quantified at landscape scale. Agricultural intensity on the surveyed farms was quantified by mean farm stocking rate, calculated as livestock units per ha (LU/ha), and generalised linear mixed models used to evaluate relationships between stocking rate and the incidence of chosen bioindicator groups. Field scale bioindicators (plant species richness and parasitoid taxon richness and abundance) were negatively associated with mean farm stocking rate. Over much of its observed range, mean farm stocking rate was positively associated with total bird species richness and abundance, and the species richness and abundance of farmland bird indicator species recorded in the winter season. However, these relationships were quadratic, and above a relatively high upper limit of 2.5–3.5 LU/ha, further increase in farm stocking rate had a negative influence. Results demonstrate that different bioindicators measured at different spatial scales vary in their response to agricultural intensity. The lack of a consistent bioindicator response to farm stocking rate suggests that within predominantly farmed regions, maximising biodiversity requires a careful targeting and monitoring with bioindicator taxa that are informative of influences at relevant operational scales. The insights provided may then be much more informative for the design and implementation of agri-environment measures that maximise biodiversity within farmed landscapes. DG 19/11/2012

Persistence of microorganisms or reinfections are the main reasons for failure of root canal therapy. Very few studies to date have included culture-independent methods to assess the microbiota, including non-cultivable microorganisms. The aim of this study was to combine culture methods with culture-independent cloning methods to analyze the microbial flora of root-filled teeth with periradicular lesions. Twenty-one samples from previously root-filled teeth were collected from patients with periradicular lesions. Microorganisms were cultivated, isolated and biochemically identified. In addition, ribosomal DNA of bacteria, fungi and archaea derived from the same samples was amplified and the PCR products were used to construct clone libraries. DNA of selected clones was sequenced and microbial species were identified, comparing the sequences with public databases. Microorganisms were found in 12 samples with culture-dependent and -independent methods combined. The number of bacterial species ranged from 1 to 12 in one sample. The majority of the 26 taxa belonged to the phylum Firmicutes (14 taxa), followed by Actinobacteria, Proteobacteria and Bacteroidetes. One sample was positive for fungi, and archaea could not be detected. The results obtained with both methods differed. The cloning technique detected several as-yet-uncultivated taxa. Using a combination of both methods 13 taxa were detected that had not been found in root-filled teeth so far. Enterococcus faecalis was only detected in two samples using culture methods. Combining the culture-dependent and –independent approaches revealed new candidate endodontic pathogens and a high diversity of the microbial flora in root-filled teeth with periradicular lesions. Both methods yielded differing results, emphasizing the benefit of combined methods for the detection of the actual microbial diversity in apical periodontitis.

Botanical and arthropod surveys at field level, and bird counts within field boundaries were undertaken on the same random sample of 50 grass-based farms in SE Ireland. Additional data relating to farm system, farm-level nutrient inputs, stocking rates, and participation (or otherwise) in the Irish Rural Environment Protection Scheme (REPS) were collated. Generalized linear models (GLM) showed that farm system was a predominant influence explaining observed biological diversity. Both sward plant and arthropod diversity were greater on non-dairy (drystock) farms, but total arthropod abundance was greater on dairy farm swards. Both the breeding bird abundance and species richness were significantly greater in field boundaries on dairy, compared with non-dairy farms. These insights have relevance to the debate regarding the most effective use of public expenditure on agri-environment policy, and suggest that such incentive schemes need to become more clearly customised to realise the conservation potential of different farming systems. DG 19/11/12