This work explores the usefulness of fish bile analysis in combination with biomarkers for identifying and evaluating new environmental contaminants in the aquatic environment. It illustrates how bile analysis can be used together with biomarkers to assess the causes of estrogenic effects, to identify chemicals in the aquatic environment that are taken up by fish, and to monitor environmental exposure. In a first application, fish exposed to sewage treatment plant effluent were studied. Elevated levels of vitellogenin in the exposed fish demonstrated that estrogenic effects occurred. Several estrogen disrupting substances were identified in the fish bile, and analysis of water samples confirmed that these substances were present in the effluent. The synthetic estrogen 17a-ethinylestradiol, which is known to be present in sewage treatment plant effluent, was shown for the first time to be taken up by fish. Considering the reported potencies of the detected substances, it was concluded that 17a-ethinylestradiol was the major contributor to the estrogenic effects. Chemical analysis of bile was used to identify rubber additives that were released from tires immersed in water. The bile of rainbow trout held in the water contained high levels of metabolites of PAHs and aromatic nitrogen compounds. Several biomarkers were also measured in the exposed fish, and EROD induction and oxidative stress were observed. Based on the bile analysis observations together with knowledge of toxicological mechanisms, it was postulated that the EROD induction was due to the PAHs, while aromatic nitrogen compounds caused the oxidative stress. Resin acids in fish bile proved to be a good indicator of exposure in a chronic long-term study of rainbow trout exposed to effluent from a total chlorine free (TCF) pulp mill. Elevated levels of GST (gluthatione-S-transferase) and GR (gluthatione reductase) activity, and the presence of DNA adducts after a two month recovery period, indicated that compounds in the pulp mill effluents have persistent effects. In addition to characterising the exposure of the fish to the effluent, the analysis of the resin acids in the bile provided evidence of accidents in the pulp mill that the existing process monitoring system had not detected. Resin acids in bile were also found to be a valuable indicator of exposure to pulp mill effluents for eelpout living in the Baltic Sea. A correlation between resin acid levels in bile and skewed sex ratios provided an important link in the chain of evidence that substances in the pulp mill effluents cause male bias of the eelpout embryos. A particularly good example of the potential of bile analysis was the identification of a previously unknown environmental contaminant. A large peak was observed in the bile extracts of fish that had been exposed to sewage treatment plant effluent. This peak was identified as triclosan, which demonstrated its presence in sewage treatment plant effluent. Other work went on to show that it is a common contaminant of the aquatic environment. The ability of fish to concentrate contaminant metabolites in bile to levels very much higher than in the environment, and the comparatively low levels of analytic interferences, make bile a particularly attractive matrix to search for new, unknown organic pollutants

OBJECTIVE: To measure urinary triclosan levels and their variability across pregnancy, and to identify sources of triclosan exposure among Canadian pregnant women. METHODS: Single spot and serial urine samples, as well as consumer product use information were collected across pregnancy and post-partum from 80 healthy pregnant women in Ottawa. Analyses included descriptives, linear mixed effects and parametric trend modeling, and surrogate category analysis. RESULTS: Triclosan was detected in 87% of maternal urine samples (LOD=3.0 µg/L). Triclosan concentrations varied by time of day of urine collection (p=0.0006), season of sampling (p=0.019), and parity (p=0.038). Triclosan was included in 4% of all personal care products used by participants; 89% of these triclosan products were varying brands of toothpaste and hand soaps. CONCLUSION: This study provided the first data on temporal variability urinary triclosan levels, and on source attribution data in Canadian pregnant women. Results will assist with population-specific exposure assessment strategies.

Pharmaceutical and personal care products (PPCPs) triclosan and nonylphenol, were investigated throughout wastewater treatment in a publicly owned treatment works (POTW). Both compounds react quickly upon chlorination under laboratory conditions, transforming into mono and dichlorinated species. A novel quantitative analytical method employing mass spectrometry was demonstrated on Delaware POTW wastewater samples. Specific transformation products were not detected and the concentration of precursor analytes was not found to be statistically different after treatment. Under tertiary chlorination conditions, transformation products are not produced. ESI-MS was used to explore triply charged, highly hydrated lanthanide ions and charge reduction was directly observed in the MS collision cell. This process proceeded via proton transfer, proved by a strong correlation between the minimum number of water molecules required to stabilize the Ln3+ and the first hydrolysis constant (R2=0.92). The effect of different solvents on the surface activity of ions under electrospray ionization (ESI) was investigated using dilute ionic liquids and the relative surface activity of a given pair of ions could be reversed by moving from a relatively polar solvent to a relatively non-polar one. / Graduate

Adult Oral Health Programme: The Effect of Periodontal Treatment and the Use of a Triclosan Containing Toothpaste on Glycaemic Control in Diabetics Abstract Aim: The aim of the research study is to establish an adult oral health programme for diabetics in Majuro, Republic of the Marshall Islands in order to determine the impact of non-surgical periodontal treatment followed by the use of a triclosan containing dentifrice on the maintenance of periodontal health and glycaemic control in type 2 diabetic patients. Hypothesis: Non-surgical periodontal treatment results in improved periodontal health and better glycaemic control in diabetics and use of a triclosan containing toothpaste is effective in maintaining this improvement in diabetics. Methods: An adult oral health programme was created, within which was conducted a two-group randomised clinical trial to address the hypothesis that non-surgical periodontal treatment results in improved periodontal health and better glycaemic control in type 2 diabetics and that the use of a triclosan containing toothpaste is effective in maintaining this improvement in diabetics. In this double blind controlled trial, sixty adult patients (aged 35 to 65 years) with type 2 diabetes mellitus having a minimum of 16 teeth received non-surgical periodontal treatment. Half of the patients were randomly assigned to use a triclosan containing toothpaste, Colgate Total, and the other group a non-triclosan toothpaste, Colgate Fluoriguard. The study evaluated the improvement in periodontal health by recording Probing Pocket Depth (PPD) on 6 sites of each tooth, and the number of sites bleeding on probing (BOP) at baseline, and at 6 months and 12 months after treatment. The second part of the study evaluated the impact of improvement of periodontal health on glycaemic control in type 2 diabetics by measuring HbA1c and RBS, and also assessing the levels of C-Peptides and CRP at baseline, and at 6 months and 12 months after treatment. The study also evaluated the effectiveness of a triclosan containing toothpaste in maintaining the improvement in periodontal health after non-surgical periodontal treatment. Results: The results showed that it was feasible to establish an oral health programme for the diabetics and could improve their periodontal health, and that toothpaste containing triclosan is effective in maintaining the improved periodontal heath in type 2 diabetics. Mean PPD dropped from 2.35mm to 1.95mm in the triclosan group and from 2.49mm to 2.24 mm in the non-triclosan group and the mean number of BOP sites dropped from 4.9 to 2.8 in the triclosan group and from 4.7 to 3.2 in the fluoriguard at 12 month visits. However, the results did not show improvement of HbA1c nor RBS levels in either group. C-Peptide levels increased and C-Reactive Protein levels decreased in both groups, however, not to significant levels at 12 month visits. Conclusion: The results of this research study lead to the conclusion that treating periodontal infection has effect of periodontal health of type 2 diabetic patients and following-up with simple personal oral hygiene of regular tooth-brushing helps maintain their periodontal health. This programme also proved that this type of oral health programme is feasible and valuable for diabetics in isolated places like the Marshall Islands, where infrastructure, personnel and resources are limited to treat microvascular and macrovascular complications of diabetes. As for the effectiveness of treating periodontal infections on glycaemic control of diabetics, this study failed to support the hypothesis that non-surgical treatment plus triclosan containing toothpaste would lead to better glycaemic management through improved periodontal health.

A maioria das pessoas não consegue desenvolver em forma adequada, a remoção total do biofilme por médios mecânicos. Em decorrência a complementação com coadjuvantes químicos disponibilizados através de cremes dentais resulta em uma opção valida. O triclosan demonstrou ser efetivo na redução dos depósitos de biofilme supragengival em estudos clínicos com ausência de remoção mecânica por até quatro dias. Contudo, não se sabe o impacto dessa medida na inibição da formação do biofilme subgengival. O objetivo do presente estudo foi comparar o efeito inibitório sobre a formação inicial do biofilme subgengival de uma suspensão de dentifrício contendo 0,3% de Triclosan e 2% de Copolimero com uma suspensão control sem triclosan. Para tanto um ensaio clínico controlado cruzado, randomizado, duplo-cego com quatro dias de duração sem medidas de higiene mecânica foi desenvolvido. Vinte e Seis indivíduos completaram os dois períodos previstos com uma fase de “wash-out” de 10 dias entre os dois. Os indivíduos bochecharam por um minuto com as suspensões a cada 12 horas. Ao longo desse período a presença da zona livre de placa foi registrada a cada 24 horas por um examinador calibrado. A formulação teste inibiu de forma significativa a formação do biofilme subgengival por até 72 horas. Pode-se concluir que bochechos diários com uma suspensão de dentifrício contendo triclosan inibiu de forma significativa a formação inicial do biofilme subgengival por um período de até 72 h comparado com um controle. / Daily systematic removal of dental biofilm in the vast majority of people does not prevent gingivitis and periodontits. Therefore the complement with chemical products delivered through toothpastes results in promising option. Triclosan has been shown effective in reducing supragingival biofilm on 4-days without tooth brushing, but unknown the impact on inhibition of biofilm subgingival. The aim of this study was to compare the inhibitory effect on the early formation of subgingival biofilm from a slurrie of toothpaste containing 0.3% Triclosan and 2% Copolymer vs. control slurrie without triclosan. A randomized crossover controlled double-blind clinical trial of 4-day period without mechanical hygiene was developed. 26 subjects completed 2 periods with wash-out period between them. People were rinsed with slurries at 12hs. A trained examiner evaluated every 24 hours, the plaque-free zone. In conclusion the triclosan/copolymer retards the early formation of biofilm subgingival up to 72 hours.

A maioria das pessoas não consegue desenvolver em forma adequada, a remoção total do biofilme por médios mecânicos. Em decorrência a complementação com coadjuvantes químicos disponibilizados através de cremes dentais resulta em uma opção valida. O triclosan demonstrou ser efetivo na redução dos depósitos de biofilme supragengival em estudos clínicos com ausência de remoção mecânica por até quatro dias. Contudo, não se sabe o impacto dessa medida na inibição da formação do biofilme subgengival. O objetivo do presente estudo foi comparar o efeito inibitório sobre a formação inicial do biofilme subgengival de uma suspensão de dentifrício contendo 0,3% de Triclosan e 2% de Copolimero com uma suspensão control sem triclosan. Para tanto um ensaio clínico controlado cruzado, randomizado, duplo-cego com quatro dias de duração sem medidas de higiene mecânica foi desenvolvido. Vinte e Seis indivíduos completaram os dois períodos previstos com uma fase de “wash-out” de 10 dias entre os dois. Os indivíduos bochecharam por um minuto com as suspensões a cada 12 horas. Ao longo desse período a presença da zona livre de placa foi registrada a cada 24 horas por um examinador calibrado. A formulação teste inibiu de forma significativa a formação do biofilme subgengival por até 72 horas. Pode-se concluir que bochechos diários com uma suspensão de dentifrício contendo triclosan inibiu de forma significativa a formação inicial do biofilme subgengival por um período de até 72 h comparado com um controle. / Daily systematic removal of dental biofilm in the vast majority of people does not prevent gingivitis and periodontits. Therefore the complement with chemical products delivered through toothpastes results in promising option. Triclosan has been shown effective in reducing supragingival biofilm on 4-days without tooth brushing, but unknown the impact on inhibition of biofilm subgingival. The aim of this study was to compare the inhibitory effect on the early formation of subgingival biofilm from a slurrie of toothpaste containing 0.3% Triclosan and 2% Copolymer vs. control slurrie without triclosan. A randomized crossover controlled double-blind clinical trial of 4-day period without mechanical hygiene was developed. 26 subjects completed 2 periods with wash-out period between them. People were rinsed with slurries at 12hs. A trained examiner evaluated every 24 hours, the plaque-free zone. In conclusion the triclosan/copolymer retards the early formation of biofilm subgingival up to 72 hours.

A maioria das pessoas não consegue desenvolver em forma adequada, a remoção total do biofilme por médios mecânicos. Em decorrência a complementação com coadjuvantes químicos disponibilizados através de cremes dentais resulta em uma opção valida. O triclosan demonstrou ser efetivo na redução dos depósitos de biofilme supragengival em estudos clínicos com ausência de remoção mecânica por até quatro dias. Contudo, não se sabe o impacto dessa medida na inibição da formação do biofilme subgengival. O objetivo do presente estudo foi comparar o efeito inibitório sobre a formação inicial do biofilme subgengival de uma suspensão de dentifrício contendo 0,3% de Triclosan e 2% de Copolimero com uma suspensão control sem triclosan. Para tanto um ensaio clínico controlado cruzado, randomizado, duplo-cego com quatro dias de duração sem medidas de higiene mecânica foi desenvolvido. Vinte e Seis indivíduos completaram os dois períodos previstos com uma fase de “wash-out” de 10 dias entre os dois. Os indivíduos bochecharam por um minuto com as suspensões a cada 12 horas. Ao longo desse período a presença da zona livre de placa foi registrada a cada 24 horas por um examinador calibrado. A formulação teste inibiu de forma significativa a formação do biofilme subgengival por até 72 horas. Pode-se concluir que bochechos diários com uma suspensão de dentifrício contendo triclosan inibiu de forma significativa a formação inicial do biofilme subgengival por um período de até 72 h comparado com um controle. / Daily systematic removal of dental biofilm in the vast majority of people does not prevent gingivitis and periodontits. Therefore the complement with chemical products delivered through toothpastes results in promising option. Triclosan has been shown effective in reducing supragingival biofilm on 4-days without tooth brushing, but unknown the impact on inhibition of biofilm subgingival. The aim of this study was to compare the inhibitory effect on the early formation of subgingival biofilm from a slurrie of toothpaste containing 0.3% Triclosan and 2% Copolymer vs. control slurrie without triclosan. A randomized crossover controlled double-blind clinical trial of 4-day period without mechanical hygiene was developed. 26 subjects completed 2 periods with wash-out period between them. People were rinsed with slurries at 12hs. A trained examiner evaluated every 24 hours, the plaque-free zone. In conclusion the triclosan/copolymer retards the early formation of biofilm subgingival up to 72 hours.

Magister Scientiae Dentium - MSc(Dent) / Aim: To determine whether there was any significant difference in the antimicrobial activity of 4 herbal toothpastes against cultures of 3 primary plaque colonizers (Streptococcus mutans, Streptococcus sanguinis and a non-specific alpha;-heamolytic streptococcus). / South Africa

OBJECTIVE: To measure urinary triclosan levels and their variability across pregnancy, and to identify sources of triclosan exposure among Canadian pregnant women. METHODS: Single spot and serial urine samples, as well as consumer product use information were collected across pregnancy and post-partum from 80 healthy pregnant women in Ottawa. Analyses included descriptives, linear mixed effects and parametric trend modeling, and surrogate category analysis. RESULTS: Triclosan was detected in 87% of maternal urine samples (LOD=3.0 µg/L). Triclosan concentrations varied by time of day of urine collection (p=0.0006), season of sampling (p=0.019), and parity (p=0.038). Triclosan was included in 4% of all personal care products used by participants; 89% of these triclosan products were varying brands of toothpaste and hand soaps. CONCLUSION: This study provided the first data on temporal variability urinary triclosan levels, and on source attribution data in Canadian pregnant women. Results will assist with population-specific exposure assessment strategies.

N-acyl ethanolamines (NAEs) are a class of lipids recently recognized as signaling molecules which are controlled, in part, by their degradation by fatty acid amide hydrolase (FAAH). On the basis of previous studies indicating increased NAE levels in a tobacco cell suspension-xylanase elicitor exposure system and the availability of FAAH mutants, overexpressor and knockout (OE and KO) genotypes in Arabidopsis thaliana, further roles of NAEs in A. thaliana plant defense was investigated. The commonly occurring urban antimicrobial contaminant triclosan (TCS) has been shown to suppress lipid signaling associated with plant defense responses. Thus, a second objective of this study was to determine if TCS exposure specifically interferes with NAE levels. No changes in steady state NAE profiles in A. thaliana-Pseudomonas syringae pv. syringae and A. thaliana-flagellin (bacterial peptide, flg22) challenge systems were seen despite evidence that defense responses were activated in these systems. There was a significant drop in enoyl-ACP reductase (ENR) enzyme activity, which catalyzes the last step in the fatty acid biosynthesis pathway in plants, on exposure of the seedlings to TCS at 10 ppm for 24 h and decreased reactive oxygen species (ROS) production due to flg22 in long term exposure of 0.1 ppm and short term exposure of 5 ppm. However, these responses were not accompanied by significant changes in steady state NAE profiles.