The oral microbiota is a reservoir for antimicrobial resistance: resistome and phenotypic resistance characteristics of oral biofilm in health, caries, and periodontitis.

BACKGROUND: Antimicrobial resistance (AMR) is an ever-growing threat to modern medicine and, according to the latest reports, it causes nearly twice as many deaths globally as AIDS or malaria. Elucidating reservoirs and dissemination routes of antimicrobial resistance genes (ARGs) are essential in fighting AMR. Human commensals represent an important reservoir, which is underexplored for the oral microbiota. Here, we set out to investigate the resistome and phenotypic resistance of oral biofilm microbiota from 179 orally healthy (H), caries active (C), and periodontally diseased (P) individuals (TRN: DRKS00013119, Registration date: 22.10.2022). The samples were analysed using shotgun metagenomic sequencing combined, for the first time, with culture technique. A selection of 997 isolates was tested for resistance to relevant antibiotics. RESULTS: The shotgun metagenomics sequencing resulted in 2,069,295,923 reads classified into 4856 species-level OTUs. PERMANOVA analysis of beta-diversity revealed significant differences between the groups regarding their microbiota composition and their ARG profile. The samples were clustered into three ecotypes based on their microbial composition. The bacterial composition of H and C samples greatly overlapped and was based on ecotypes 1 and 2 whereas ecotype 3 was only detected in periodontitis. We found 64 ARGs conveying resistance to 36 antibiotics, particularly to tetracycline, macrolide-lincosamide-streptogramin, and beta-lactam antibiotics, and a correspondingly high prevalence of phenotypic resistance. Based on the microbiota composition, these ARGs cluster in different resistotypes, and a higher prevalence is found in healthy and caries active than in periodontally diseased individuals. There was a significant association between the resistotypes and the ecotypes. Although numerous associations were found between specific antibiotic resistance and bacterial taxa, only a few taxa showed matching associations with both genotypic and phenotypic analyses. CONCLUSIONS: Our findings show the importance of the oral microbiota from different niches within the oral cavity as a reservoir for antibiotic resistance. Additionally, the present study showed the need for using more than one method to reveal antibiotic resistance within the total oral biofilm, as a clear mismatch between the shotgun metagenomics method and the phenotypic resistance characterization was shown.

Effect of xylitol on salivary ß-glucosidase in humans.

Dental biofilm - in which a diverse set of microorganisms are embedded in a complex polysaccharide matrix that adheres to oral components - is one of the most complex microbial communities in the human body. As biofilm formation is related to oral infections, such as caries and periodontal diseases, strategies for biofilm control are crucial for maintaining oral health. Xylitol, a synthetic sugar used as a sucrose substitute, has been shown to reduce biofilm formation. However, its precise mechanism of action on biofilm reduction has so far not been elucidated. Previous studies demonstrate that bacterial ß-glucosidase action is crucial for biofilm formation. Here, we investigated the correlation between salivary ß-glucosidase activity and dental plaque occurrence. We found a positive correlation between enzymatic activity and the presence of dental biofilm. We observed that xylitol inhibits ß-glucosidase in human saliva. Kinetic studies also confirmed that xylitol acts as a mixed type inhibitor of salivary ß-glucosidase. Based on our data, we suggest that xylitol impairs oral biofilm formation by the inhibition of bacterial ß-glucosidase, which is essential for biofilm formation in the oral cavity.

Supplementary sampling of obturation materials enhances microbial analysis of endodontic treatment failures: a proof of principle study.

OBJECTIVES: 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. MATERIALS AND METHODS: 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. RESULTS: 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 × 10(3) CFU/ml). Of these, nine were detected solely in the retrieved obturation materials. The reinfected root canals harbored 13 taxa (3.48-7.36 × 10(3) 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. CONCLUSIONS: Bacteria can colonize both root canals and endodontic fillings in vivo. CLINICAL RELEVANCE: 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.

Novel Broad-Spectrum Antimicrobial Photoinactivation of In Situ Oral Biofilms by Visible Light plus Water-Filtered Infrared A.

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 log10 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.

Microscope-based imaging platform for large-scale analysis of oral biofilms.

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.

Durability of microtensile bond to nonetched and etched feldspar ceramic: self-adhesive resin cements vs conventional resin.

PURPOSE: The objective of this study was to evaluate the effect of thermocycling (TC), self-adhesive resin cements and surface conditioning on the microtensile bond strength (µTBS) between feldspathic ceramic blocks and resin cements. MATERIALS AND METHODS: Fifty-six feldspathic ceramic blocks (10 x 7 x 5 mm) (Vita Mark II) were divided into groups according to the factors "resin cement" (3 cements) and "surface conditioning" (no conditioning or conditioning [10% hydrofluoric acid etching for 5 min + silanization]) (n = 8): group 1: conditioning+Variolink II (control group); group 2: no conditioning+Biscem; group 3: no conditioning+RelyX U100; group 4: no conditioning+Maxcem Elite; group 5: conditioning+ Biscem; group 6: conditioning+RelyX U100; group 7: conditioning+Maxcem Elite. The ceramic-cement blocks were sectioned to produce non-trimmed bar specimens (adhered cross-sectional area: 1 ± 0.1 mm2), which were divided into two storage conditions: dry, µTBS immediately after cutting; TC (12,000x, 5°C/55°C). Statistical significance was determined using two-way ANOVA (7 strategies and 2 storage conditions) and the post-hoc Tukey test (p < 0.05). RESULTS: Resin cement and thermocycling affected the µTBS significantly (p = 0.001). In the dry condition, group 5 (18 ± 6.5 MPa) presented the lowest values of µTBS when compared to the other groups. TC decreased the mean µTBS values significantly (p < 0.05) for all resin cements tested (9.7 ± 2.3 to 22.1 ± 6.3 MPa), except for the resin cement RelyX U100 (22.1 ± 6.3 MPa). In groups 3 and 4, it was not possible to measure µTBS, since these groups had 100% pre-test failures during sectioning. Moreover, the same occurred in group 2 after TC, where 100% failure was observed during thermocycling (spontaneous failures). CONCLUSION: Hydrofluoric acid etching and silanization of the feldspathic ceramic surface are essential for bonding self-adhesive resin cement to a feldspathic ceramic, regardless of the resin cement used. Non-etched ceramic is not recommended.

Antimicrobial potential of resin matrices loaded with coffee compounds.

This study evaluated the biological behavior of the coffee compounds Trigonelline (T), chlorogenic acid (C), and nicotinic acid (N), correlating with their release from a resin matrix. Minimum inhibitory concentration (MIC) was evaluated against Streptococcus mutans UA159, and cytotoxicity was assessed by methyl tetrazolium salt on OD-21 cells. Resin matrices (bisphenol A-glycidyl-dimethacrylate/triethylene glycol-dimethacrylate 70/30 wt%, camphorquinone/ethyl 4-dimethyl aminobenzoate 0.5/1 wt%) were doped with coffee compounds in different concentrations (10/20/30/40/50 wt%), performing 15 groups (T10-T50, C10-C50, N10-N50), and a control group with no coffee compound. Degree of conversion (DC%) was analyzed by Fourier transform infrared spectroscopy. Antimicrobial properties were evaluated by bioluminescence (Luciferase assay). The release from loaded matrices was analyzed over time (24 hr, 6, 14, 21 and 28 days), using high-performance liquid chromatography (HPLC). Data were submitted to ANOVA/Tukey's test (α = 0.05). MIC for T and C was 6 mg/ml, and 4 mg/ml for N. None of them were cytotoxic. Only T50 and C50 showed lower DC% than control (α < 0.05). Some groups (T30/T40/T50/C40/C50/N50) were strongly antimicrobial, reducing bacterial activity approximately five times compared to control (α < 0.05). For T30, T40, T50, C40, and C50, the HPLC showed a release above or closer to MIC values mainly in 24 hr, but for N50, up to 28 days. In conclusion, the coffee compounds presented antimicrobial activity, depending on their concentration when added in resin matrices, being found a correlation with their release.

Influence of the long-term use of oral hygiene products containing stannous ions on the salivary microbiome - a randomized controlled trial.

Oral hygiene products containing tin are suitable to prevent erosive tooth wear, yet effects on the oral microbiota are not known yet. Therefore, this study determined the salivary microbiome of 16 participants using products with stannous ions for three years (TG) compared with a control group (CG) to assess their influence on the microbiota. Participants were included in a randomized controlled clinical trial (RCT) with biannual visits. Illumina Miseq sequencing revealed as most abundant genera: Streptococcus (TG 14.3%; CG 13.0%), Veillonella (TG 11.3%; CG 10.9%), Prevotella (TG 7.0%; CG 9.8%), Haemophilus (TG 6.6%; CG 7.2%), Porphyromonas (TG 5.9%, CG 5.1%), Leptotrichia (TG 5.8%; CG 4.9%), Actinomyces (TG 4.0%; CG 4.6%) and Neisseria (TG 5.4%; CG 4.2%). Beta-Diversity was not significantly different between groups at both time points, although significant differences between groups were found for certain taxa after three years. The genus Prevotella was found in higher abundance in CG whereas Neisseria and Granulicatella, health-associated taxa, were found more abundantly in TG. Salivary microbiota after three years reflected a composition associated with oral health, hence continual use as a preventive measure for dental erosion can be considered safe and benefitting oral health for patients with a high risk of erosion.

A comparison of the film thickness of two adhesive luting agents and the effect of thermocycling on their microTBs to feldspathic ceramic.

The aim of the present study was to evaluate the effect of thermocycling (TC) on the microtensile bond strength (microTBS) of two luting agents to feldspathic ceramic and to measure their film thickness (FT). For the microTBS test, sixteen blocks (6.4 x 6.4 x 4.8 mm) were fabricated using a feldspathic ceramic, etched with 10% hydrofluoric acid, rinsed and treated with the silane agent. The ceramic blocks were divided into two groups (n= 8): Gr1: dual-cured resin cement and Gr2: flowable resin. The luting agents were applied on the treated surfaces. Microsticks (1 +/-0. 1mm2) were prepared and stored under two conditions: dry, specimens immediately submitted to the microTBS test, and TC (6,000 cycles; 5 degrees C-55 degrees C). The microTBS was evaluated using a universal testing machine (1 mm/min). The microTBS data (MPa) were submitted to two-way ANOVA and Tukey' test (5%). For the FT test (ISO 4049), 0.05 ml of each luting agent (n=8) was pressed between two Mylar-covered glass plates (150 N) for 180 seconds and light polymerized. FT was measured using a digital paquimeter (Model 727-2001). The data (mm) were submitted to one-way ANOVA. The luting cement did not influence the microTBS results (p= 0.4467). Higher microtensile bond values were found after TC (20.5 +/- 8.6 MPa) compared to the dry condition (13.9 +/- 4. 7MPa), for both luting agents. The luting agents presented similar film thicknesses: Gr1- 0.052 +/- 0.016 mm; Gr2-0.041 +/- 0.003 mm. The luting agents presented similar film thickness and microTBS values, in dry and TC conditions and TC increased the bond strength regardless of the luting agent.

Batch injection analysis towards auxiliary diagnosis of periodontal diseases based on indirect amperometric detection of salivary α-amylase on a cupric oxide electrode.

This study describes, for the first time, the use of a batch injection analysis system with amperometric detection (BIA-AD) to indirectly determine salivary α-amylase (sAA) levels in saliva samples for chronic periodontitis diagnosis. A chemical/thermal treatment was explored to generate a CuO film on a Cu electrode surface. This procedure offered good stability (RSD = 0.3%), good repeatability (RSD < 1.3%) and excellent reproducibility (RSD < 1.5%). The sAA concentration levels were determined based on the detection of maltose produced by enzymatic hydrolysis of starch. The analytical performance was investigated, and a linear correlation was observed for a maltose concentration range between 0.5 and 6.0 mmol L-1 with a correlation coefficient equal to 0.999. The analytical sensitivity and the limit of detection were 48.8 µA/(mmol L-1) and 0.05 mmol L-1, respectively. In addition, the proposed system provided an excellent analytical frequency (120 analysis h-1). The clinical feasibility of the proposed method was investigated by the determination of sAA levels in four saliva samples (two from healthy control persons (C1 and C2) and two from patients with chronic periodontitis (P1 and P2)). The accuracy provided by the BIA-AD system ranged from 93 to 98%. The sAA concentration levels achieved for each sample were compared to the values found by spectrophotometry and there was no statistically significant difference between them at a confidence level of 95%. Finally, the method reported herein emerges as a simple, low cost and promising tool for assisting periodontal diseases diagnosis.

Microplastics in personal care products: Exploring perceptions of environmentalists, beauticians and students.

Microplastics enter the environment as a result of larger plastic items breaking down ('secondary') and from particles originally manufactured at that size ('primary'). Personal care products are an important contributor of secondary microplastics (typically referred to as 'microbeads'), for example in toothpaste, facial scrubs and soaps. Consumers play an important role in influencing the demand for these products and therefore any associated environmental consequences. Hence we need to understand public perceptions in order to help reduce emissions of microplastics. This study explored awareness of plastic microbeads in personal care products in three groups: environmental activists, trainee beauticians and university students in South West England. Focus groups were run, where participants were shown the quantity of microbeads found in individual high-street personal care products. Qualitative analysis showed that while the environmentalists were originally aware of the issue, it lacked visibility and immediacy for the beauticians and students. Yet when shown the amount of plastic in a range of familiar everyday personal care products, all participants expressed considerable surprise and concern at the quantities and potential impact. Regardless of any perceived level of harm in the environment, the consensus was that their use was unnatural and unnecessary. This research could inform future communications with the public and industry as well as policy initiatives to phase out the use of microbeads.

Combination coatings unlock medical device potential.

Employing combination coatings, whether multiple coatings on the same surface or different coatings on different parts of the same device offer medical devices a number of benefits. Examples of what can be achieved are reported here.

Antimicrobial Photoinactivation Using Visible Light Plus Water-Filtered Infrared-A (VIS + wIRA) Alters In Situ Oral Biofilms.

Recently, growing attention has been paid to antimicrobial photodynamic therapy (aPDT) in dentistry. Changing the microbial composition of initial and mature oral biofilm by aPDT using visible light plus water-filtered infrared-A wavelengths (VIS + wIRA) has not yet been investigated. Moreover, most aPDT studies have been conducted on planktonic bacterial cultures. Therefore, in the present clinical study we cultivated initial and mature oral biofilms in six healthy volunteers for 2 hours or 3 days, respectively. The biofilms were treated with aPDT using VIS+wIRA (200 mW cm(-2)), toluidine blue (TB) and chlorine e6 (Ce6) for 5 minutes. Chlorhexidine treated biofilm samples served as positive controls, while untreated biofilms served as negative controls. After aPDT treatment the colony forming units (CFU) of the biofilm samples were quantified, and the surviving bacteria were isolated in pure cultures and identified using MALDI-TOF, biochemical tests and 16S rDNA-sequencing. aPDT killed more than 99.9% of the initial viable bacterial count and 95% of the mature oral biofilm in situ, independent of the photosensitizer. The number of surviving bacterial species was highly reduced to 6 (TB) and 4 (Ce6) in the treated initial oral biofilm compared to the 20 different species of the untreated biofilm. The proportions of surviving bacterial species were also changed after TB- and Ce6-mediated aPDT of the mature oral biofilm, resulting in a shift in the microbial composition of the treated biofilm compared to that of the control biofilm. In conclusion, aPDT using VIS + wIRA showed a remarkable potential to eradicate both initial and mature oral biofilms, and also to markedly alter the remaining biofilm. This encourages the clinical use of aPDT with VIS + wIRA for the treatment of periimplantitis and periodontitis.

Factors related to tracking of blood pressure in children. U.S. National Center for Health Statistics Health Examination Surveys Cycles II and III.

In this paper we examine the relationship of growth, obesity, and the degree of sexual and bone maturation to blood pressure in a U.S. national probability sample of 2165 children examined by the U.S. National Center for Health Statistics on two occasions, approximately 4 years apart. Subjects who maintained, increased, or decreased their peer rank order of blood pressure are described. Children who maintained their blood pressure in the upper quintile were taller, heavier, more obese, had greater bone age, greater numbers of permanent teeth, and were more sexually mature than their peers, while those maintaining their blood pressure in the lowest quintile of blood pressure were shorter, lighter, less obese, had lesser bone age, fewer permanent teeth, and were less sexually mature. Subjects whose blood pressures were initially in the lowest four quintiles and then rose to the top quintile were also taller, heavier, more obese, and had greater bone age, while those with blood pressures falling to the lowest quintile from the upper four quintiles were shorter, lighter, less obese, and had lesser bone age. Thus, the level at which blood pressure tracks during childhood is related to growth, obesity, and to the degree of maturation acquired. In addition, children whose blood pressures are rising or falling in relation to their peers have body growth and maturation characteristics similar to those who maintain their rank order high or low respectively.

Platelet deposition and fibrinogen binding on surfaces coated with heparin or friction-reducing polymers.

The blood-contacting properties of polyethylene coated with a lubricious hydrophilic coating; an uncoated polyethylene; or a photoheparin-treated polyethylene-negative control sample were compared by measuring fibrinogen adsorption, antifibrinogen binding, and platelet attachment from human plasma. The polyethylene surfaces coated with a hydrophilic polymer were found to be similar to surfaces coated with heparin. Fibrinogen adsorption on the hydrophilic coating was 60% lower than adsorption on either the uncoated or heparin-coated polyethylene samples. Antifibrinogen binding from buffer to the hydrophilic coating was also reduced more than 85% from binding to uncoated polyethylene samples. Both the hydrophilic coating and heparin coating showed a reduction in platelet attachment by a factor of 100 over the uncoated sample as well as significantly reduced platelet activation.

Estrogen receptors and growth response in cultured human periodontal ligament cells.

The periodontal ligament (PDL) is a connective tissue involved in the remodeling process associated with tooth development and positioning. PDL cells grown in culture were analyzed for the capacity to specifically bind steroid hormones and for growth response to estradiol-17 beta. Using [3H]estradiol-17 beta as the ligand, PDL cells in first passage cultures exhibited a specific estrogen binding capacity of 881 fmol/mg cell protein. With [3H]dexamethasone as a ligand, the binding capacity of the glucocorticoid receptor was 143 fmol/mg protein. With [3H]R5020 as a ligand, the progestin receptor exhibited a binding capacity of 5 pmol/mg protein. Scatchard analysis of estradiol binding at 37 degrees revealed a dissociation constant of 2.7 X 10(-9) M, representative of the estrogen receptor. The addition of estradiol-17 beta at concentrations of 10(-9) and 10(-8) M to culture media induced a dose-dependent decrease in growth (DNA content) to 62% and 38% control values, respectively. The addition of the antiestrogens tamoxifen and 4-hydroxytamoxifen at concentrations of 10(-7) and 10(-6) M similarly depressed cell growth. These results show that PDL cells contain high affinity receptors for several steroid hormones and further that these cells are targets for the action of estrogens.

Chain termination in polyhydroxyalkanoate synthesis: involvement of exogenous hydroxy-compounds as chain transfer agents.

We have identified a range of compounds which, when present during poly(3-hydroxybutyrate) [P(3HB)] accumulation by Ralstonia eutropha (reclassified from Alcaligenes eutrophus), can act as chain transfer agents in the chain termination step of polymerization. End-group analysis by 31P NMR of polymer derivatized with 2-chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane revealed that all these compounds were covalently linked to P(3HB) at the carboxyl terminus. All chain transfer agents possessed one or more hydroxyl groups, and glycerol was selected for further investigation. The number-average molecular mass (Mn) of P(3HB) produced by R. eutropha from glycerol was substantially lower than for polymer produced from glucose, and we identified two new end-group structures. These were attributed to a glycerol molecule bound to the P(3HB) chain via the primary or secondary hydroxyl groups. When a primary hydroxyl group of glycerol is involved in chain transfer, the end-group structure is in both [R] and [S] configurations, implying that chain transfer to glycerol is a random transesterification and that PHA synthase does not catalyse chain transfer. 3-Hydroxybutyric acid is the most probable chain transfer agent in vivo, with propagation and termination reactions involving transfer of the P(3HB) chain to enzyme-bound and free 3-hydroxybutyrate, respectively. Only carboxyl end-groups were detected in P(3HB) extracted from exponentially growing bacteria. It is proposed that a compound other than 3-hydroxybutyryl-CoA acts as a primer in the initiation of polymer synthesis.

Biosynthesis and composition of bacterial poly(hydroxyalkanoates).

It is well established that Alcaligenes eutrophus can accumulate a copolymer containing 3-hydroxybutyrate and 3-hydroxyvalerate, but longer 3-hydroxyacid monomers have not been reported to occur in this organism. The properties of the enzymes of poly(hydroxyalkanoate) (PHA) biosynthesis are discussed and it is proposed that the substrate specificity of the polymerizing enzyme restricts the range of monomer units incorporated into PHA. Various other bacteria produce similar copolymers from propionic acid and/or valeric acid. A number of Pseudomonas species accumulate PHAs containing longer-chain monomer units from linear alkanoic acids, alkanes and alcohols.

Accumulation of a poly(hydroxyalkanoate) copolymer containing primarily 3-hydroxyvalerate from simple carbohydrate substrates by Rhodococcus sp. NCIMB 40126.

A number of taxonomically-related bacteria have been identified which accumulate poly(hydroxyalkanoate) (PHA) copolymers containing primarily 3-hydroxyvalerate (3HV) monomer units from a range of unrelated single carbon sources. One of these, Rhodococcus sp. NCIMB 40126, was further investigated and shown to produce a copolymer containing 75 mol% 3HV and 25 mol% 3-hydroxybutyrate (3HB) from glucose as sole carbon source. Polyesters containing both 3HV and 3HB monomer units, together with 4-hydroxybutyrate (4HB), 5-hydroxyvalerate (5HV) or 3-hydroxyhexanoate (3HHx), were also produced by this organism from certain accumulation substrates. With valeric acid as substrate, almost pure (99 mol% 3HV) poly(3-hydroxyvalerate) was produced. N.m.r. analysis confirmed the composition of these polyesters. The thermal properties and molecular weight of the copolymer produced from glucose were comparable to those of PHB produced by Alcaligenes eutrophus.

Contamination of dental unit cooling water with oral microorganisms and its prevention.

Each time the handpiece turbine is stopped while the bur is still in the patient's mouth, almost 1 ml of microbe-laden oral fluids may be aspirated into the average dental unit water line by the retraction valve present in the dental unit. This fluid may contain an average in excess of 54,000 microorganisms per milliliter, including both facultative and obligate anaerobic bacteria of medium to high virulence. Other infectious agents including hepatitis, herpes, influenza, and other upper respiratory viruses may also be present. This germ-laden water may then be sprayed into the mouth of the next patient, possibly initiating an oral or upper respiratory tract infection. Sterilizing the handpiece between appointments, although of great significance in the prevention of disease transmission, will not totally eliminate the problem discussed here as almost 95% of the oral fluid aspirated into the dental unit went past the handpiece and into the handpiece cooling water line. Complete elimination of the retraction valve in the unit is the most effective means of solving this problem, but, with present dental unit designs, this may result in water dripping onto the patient. A viable alternative is the installation of a check valve (anti-retraction valve) downstream from the retraction valve. Installation of a check valve resulted in an almost 4,000-fold decrease in this contamination. The attachment of a check valve will have no effect on normal less virulent bacteria already present in tap water. These bacteria colonize the water lines of the dental unit and are then continually shed into the water in numbers often as high as 1,000,000 CFU/ml.(ABSTRACT TRUNCATED AT 250 WORDS)