Antibiotic concentrations in raw hospital wastewater surpass minimal selective and minimum inhibitory concentrations of resistant Acinetobacter baylyi strains.

Antibiotics are essential for modern medicine, they are employed frequently in hospitals and, therefore, present in hospital wastewater. Even in concentrations, that are lower than the minimum inhibitory concentrations (MICs) of susceptible bacteria, antibiotics may exert an influence and select resistant bacteria, if they exceed the MSCs (minimal selective concentrations) of resistant strains. Here, we compare the MSCs of fluorescently labelled Acinetobacter baylyi strains harboring spontaneous resistance mutations or a resistance plasmid with antibiotic concentrations determined in hospital wastewater. Low MSCs in the µg/L range were measured for the quinolone ciprofloxacin (17 µg/L) and for the carbapenem meropenem (30 µg/L). A 24 h continuous analysis of hospital wastewater showed daily fluctuations of the concentrations of these antibiotics with distinctive peaks at 7-8 p.m. and 5-6 a.m. The meropenem concentrations were always above the MSC and MIC values of A. baylyi. In addition, the ciprofloxacin concentrations were in the range of the lowest MSC for about half the time. These results explain the abundance of strains with meropenem and ciprofloxacin resistance in hospital wastewater and drains.

[Lead in drinking water-an old problem, a new EU directive]. / Blei im Trinkwasser ­ ein altes Problem, eine neue EU-Richtlinie.

BACKGROUND: Lead pipes have been used in drinking water installations for a long time. Only since 1973 have their use in new buildings been strongly discouraged; nevertheless, they are still found in old buildings. Furthermore, lead-containing alloys are still used today in components such as fittings. This results in an avoidable pollution of drinking water. The health significance of this pollution is acknowledged by a reduction of the currently valid limit value from 10 µg/l to 5 µg/l by the new EU drinking water directive passed in 2020. This directive provides for a transition period of 15 years. OBJECTIVES: The relevance of a stricter limit value for lead in drinking water will be evaluated based by the results of routine analyses, and the necessary public health measures to prevent exposure of vulnerable groups will be formulated. MATERIAL AND METHODS: A retrospective analysis of routine samples from the city of Bonn, which an accredited drinking water laboratory had examined in the years 1997-2019, was performed. RESULTS: Of the 16,060 samples analyzed, 75.36% were below the quantitative limit of quantification. The median of the quantifiable samples was above the future limit for lead in drinking water in each year considered. No effect of the last transition period of 10 years (2003-2013) could be detected. DISCUSSION: Even though no systematic studies on lead pollution in German municipalities are available, the subject is still highly topical. Lead exposure from drinking water installations is a completely avoidable health risk, but it can only be regulated by consistent enforcement of the applicable rules by the health authorities. The health authorities must be adequately equipped in terms of personnel, material, and financial resources.

[Aluminium release of glitter particles in removable orthodontic appliances]. / Freisetzung von Aluminium aus Glitzerpartikeln bei herausnehmbaren kieferorthopädischen Apparaturen.

BACKGROUND AND AIM: In order to support children's compliance with orthodontic treatment, glitter particles containing aluminium (Al) are often embedded in the acrylic of removable appliances. When worn for up to 16 h daily for 2-3 years, it can be assumed that Al ions diffuse into saliva over time. The aim of this study was to investigate the release of Al ions from the acrylic using different orthodontic wires. MATERIALS AND METHOD: Test specimens (surface area 5.65 cm2) were prepared from orthodontic resin and various wires; half contained aluminium glitter particles. The test specimens were placed in Petri dishes containing 50 ml of corrosion medium (pH 2.3) according to DIN EN ISO 10271 at 37 °C for 7 days. Inductively coupled plasma mass spectrometry (ICP-MS) was used to quantify the specific ions in the corrosion solution. RESULTS: Statistical analysis showed a significant difference in the concentration of Al ions between samples with and without glitter particles. Concentrations from samples with glitter reached up to 14,474 µg/l Al ions; samples without glitter contained on average 1260 µg/l. A small proportion of the Al ions may originate from the alloys of the wires. CONCLUSIONS: It should be investigated whether the aluminium concentration can lead to health risks for humans. In view of the findings, orthodontists should not offer appliances containing glitter in order to minimize aluminium uptake with saliva. It needs to be clarified whether the conditions found in the oral cavity lead to the same results as under the abovementioned conditions. Legislation should be developed to limit the release of aluminium from orthodontic products.

Exposure of a single wild boar population in North Rhine-Westphalia (Germany) to perfluoroalkyl acids.

Perfluoroalkyl acids (PFAA) are among the leading chemical pollutants in the twenty-first century. Of these, perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) have been widely detected in a large number of animal and environmental samples. Wild boars accumulate PFAA in their livers, but it has not yet been clarified to what extent wild boars of the same population accumulate different PFAA in their livers or whether any conclusions can be drawn from any differences found in regard to environmental contamination. In this study, liver samples from wild boars killed during driven hunts in 2019 and 2020 from a defined forest area in North Rhine-Westfalia, Germany were analyzed for 13 different PFAA. A mean load of 493 µg/kg (± 168 µg/kg) PFAA was measured in 2020. Perfluorosulfonic acids accounted for 87% of the total load in both years, with PFOS dominating this group. These results were similar to those of 14 liver samples collected from other regions of Germany for comparison. In addition, the livers of hunted pregnant sows and fetuses were examined. The load of short-chain perfluorocarboxylic acids (< C8) in the fetus liver was as high as that of the sows, whereas the concentrations of long-chain perfluorocarboxylic acids (≥ C8) were lower than in the dams. This result shows for the first time that fetuses take up PFAA from their mothers in utero. Our study shows that PFAA content in wild boar livers is comparably high in all animals in a local population and indicates a need for further research regarding a nationwide background exposure to PFAA in wild boars and their surrounding environment.

In Vitro Ion Release of Wires in Removable Orthodontic Appliances.

Various orthodontic wire compositions and configurations are present on the market for removable appliances; however, there have still been only few studies focusing on the effect of resin color and additives such as glitter on corrosion of metallic wires under different conditions. Thus, the aim of the study was to compare concentrations of released ions (aluminium, chromium, nickel) in a corrosive medium under three different conditions: non-loaded wires, loaded wires, and non-loaded wires treated with Kukis® cleaning tablets. Six different wires made of three types of steel alloy were embedded in PMMA resin leaving one centimetre of each wire emerging from the resin to come into contact with the corrosive medium. Glitter particles were added to half of the produced test specimens. For the unloaded test series, five specimens of each group were covered in a petri dish with 50 mL of corrosive medium (pH 2.3) following EN-ISO 10271 for seven days at 37 °C. The wires for the mechanically loaded test specimens overlapped the resin by 5 cm and were clamped into a time-switched electric drive for a defined period of time before the samples were taken after a testing time of 7 days. In the third group, unloaded test specimens were transferred from their petri dishes into the prepared Kukis® solution every 24 h before being stored in the corrosive medium. Inductively coupled plasma mass spectrometry (ICP-MS) was used to quantify the specific ions in the corrosive solution. Statistical analysis showed that the mechanical loading of all wires could significantly raise the diffusion of ions into the corrosive medium. The colour of the resin did not affect the concentration of the released ions. The Kukis® cleaning tabs could not lower the corrosion of the tested metals, as some of the wires were corroded even more using the brace cleanser. Glitter-containing test specimens showed significantly higher amounts of aluminium. Mechanical loading as well as the presence of glitter particles in the resin significantly affected ion concentrations.

Assessment of a groundwater contamination with vinyl chloride (VC) and precursor volatile organic compounds (VOC) by use of a geographical information system (GIS).

Regarding the health effects of volatile organic compounds (VOC) and their decomposition products (particularly vinyl chloride (VC)) under chronic low-dose exposure, VOC groundwater contaminations are seen to be an ongoing public health issue. This article presents results of a long-term investigation surveying VOC and VC groundwater contamination upstream of a large groundwater works in Cologne, Germany. For 10 years a contaminated aquifer has been monitored for different VOC and for VC. In total, 255 samples have been taken to assess both the 3-dimensional distribution and the temporal dynamics of the contaminants. VOC and VC precursor substances have been measured by means of pentane-liquid-liquid-extraction, GC and ECD, VC by means of derivatisation to 1,2 dibromochloroethane, GC, ECD, and by purge and trap technique and GC-MS-coupling. For spatial analysis all test results and additional hydrogeological attribute data have been transferred to a GIS. The spatial VOC distribution has been assessed by use of kriging interpolation indicating a decrease of the initial contaminants in time. A cluster analysis allowed to distinguish several independent contaminations within the large contamination area. The VC contamination was increasing. Anaerobic microbial dechlorination of VOC and subsequent VC accumulation were seen to be as credible from several indications (VC presence, downstream change of tetra/trichloroethylene-ratio and anaerobic conditions in the aquifer, high Fe(2+)- and Mn(2+)-concentrations). There was no statistically significant vertical differentiation of VOC and VC concentrations. The VOC load within the different water protection zones of the waterworks could be assessed.

Perfluorooctane sulfonate increases the genotoxicity of cyclophosphamide in the micronucleus assay with V79 cells. Further proof of alterations in cell membrane properties caused by PFOS.

Perfluorooctane sulfonate (PFOS; C8F17SO3-) is a fully fluorinated organic compound which has been manufactured for decades and was used widely in industrial and commercial products. The recent toxicological knowledge of PFOS mainly concerns mono-substance exposures of PFOS to biological systems, leaving the potential interactive effects of PFOS with other compounds as an area where understanding is significantly lacking. However, a recent study, reported the potential of PFOS to enhance the toxicity of two compounds by increasing cell membrane permeability. This is of particular concern since PFOS has been reported to be widely distributed in the environment where contaminants are known to occur in complex mixtures. In this study, PFOS was evaluated alone and in combination with cyclophosphamide (CPP) to investigate whether a presence of PFOS leads to an increased genotoxic potential of CPP towards hamster lung V79 cells. Genotoxicity was investigated using the micronucleus (MN) assay according to the recent draft ISO/DIS 21427-2 method. PFOS alone demonstrated no genotoxicity up to a concentration of 12.5 microg/ml. However, PFOS combined with two different concentrations of CPP, with metabolic activation, caused a significant increase in the number of micronucleated cells compared to treatments with CPP alone. These results provide a first indication that PFOS has the potential to enhance the genotoxic action of CPP towards V79 cells, suggesting, together with the alterations in cell membrane properties shown previously, that genotoxicity of complex mixtures may be increased significantly by changes in chemical uptake. Together with an earlier study performed by the own working group, it can be concluded that PFOS alone is not genotoxic in this bioassay using V79 cells up to 12.5 microg/ml, but that further investigations are needed to assess the potential interaction between PFOS and other substances, in particular regarding the impact of membrane alterations on the uptake of toxic substances.

Perfluorinated surfactants in surface and drinking waters.

GOAL, SCOPE AND BACKGROUND: In this paper recent results are provided of an investigation on the discovery of 12 perfluorinated surfactants (PS) in different surface and drinking waters (Skutlarek et al. 2006 a, Skutlarek et al. 2006 b). In the last years, many studies have reported ubiquitous distribution of this group of perfluorinated chemicals, especially perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) in the environment, particularly in wildlife animal and human samples (Giesy and Kannan 2001, Houde et al. 2006, Prevedouros et al. 2006). Perfluorinated surfactants (e.g. PFOS and PFOA) have shown different potentials for reproductory interference and carcinogenity in animal experiments as well as partly long half-lives in humans (Guruge et al. 2006, FSA UK 2006a, FSA UK 2006b, 3M 2005, OECD 2002, Yao and Zhong 2005). They possess compound-dependent extreme recalcitrance against microbiological and chemical degradation and, in addition, they show variable potentials for bioaccumulation in animals and humans (Houde et al. 2006). METHODS: Surface and drinking water samples were collected from different sampling sites: Surface waters: samples taken from the rivers Rhine, Ruhr, Moehne and some of their tributaries. Further samples were taken from the Rhine-Herne-Canal and the Wesel-Datteln-Canal. Drinking waters: samples taken in public buildings of the Rhine-Ruhr area. After sample clean-up and concentration by solid-phase extraction, the perfluorinated surfactants were determined using HPLC-MS/MS. RESULTS: All measured concentrations (sum of seven mainly detected components) in the Rhine river and its main tributaries (mouths) were determined below 100 ng/L. The Ruhr river (tributary of the Rhine) showed the highest concentration (94 ng/L), but with a completely different pattern of components (PFOA as major component), as compared with the other tributaries and the Rhine river. Further investigations along the Ruhr river showed remarkably high concentrations of PS in the upper reaches of the Ruhr river and the Moehne river (tributary of the Ruhr) (Ruhr: up to 446 ng/L, Moehne: up to 4385 ng/L). The maximum concentration of all drinking water samples taken in the Rhine-Ruhr area was determined at 598 ng/L with the major component PFOA (519 ng/L). DISCUSSION: The surface water contaminations most likely stem from contaminated inorganic and organic waste materials (so-called 'Abfallgemisch'). This waste material was legally applied to several agricultural areas on the upper reaches of the Moehne. Perfluorinated surfactants could be detected in some suchlike soil samples. They contaminated the river and the reservoir belonging to it, likely by superficial run-off over several months or probably years. Downstream, dilution effects are held responsible for decreasing concentrations of PS in surface waters of the Moehne and the Ruhr river. In analogy to the surface water samples, PS (major component PFOA) can be determined in many drinking water samples of the Rhine-Ruhr area where the water supplies are mainly based on bank filtration and artificial recharge. CONCLUSIONS: The concentrations found in drinking waters decreased with the concentrations of the corresponding raw water samples along the flow direction of the Ruhr river (from east to west) and were not significantly different from surface water concentrations. This indicates that perfluorinated surfactants are at present not successfully removed by water treatment steps. RECOMMENDATIONS AND PERSPECTIVES: Because of their different problematic properties (persistence, mobility, toxicity, bioaccumulation), the concentrations of specific perfluorinated surfactants and their precursors in drinking waters and food have to be minimised. Therefore, it is of utmost importance to take the initiative to establish suitable legal regulations (limitations/ban) concerning the production and use of these surfactants and their precursors. Furthermore, it is indispensable to protect water resources from these compounds. A discussion on appropriate limit values in drinking water and foodstuffs is urgently needed. Concerning the assumed soil contamination, the corresponding regulation (Bioabfall-Verordnung 1998--Regulation on Organic Waste 1998) should be extended to allow the control of relevant organic pollutants.

Does perfluorooctane sulfonate (PFOS) act as chemosensitizer in zebrafish embryos?

Earlier studies have shown that perfluorooctane sulfonate (PFOS) increases the toxicity of other chemicals by enhancing their uptake by cells and tissues. The present study aimed at testing whether the underlying mechanism of enhanced uptake of chemicals by zebrafish (Danio rerio) embryos in the presence of PFOS is by interference of this compound with the cellular efflux transporter Abcb4. Modifications of uptake/clearance and toxicity of two Abcb4 substrates, the fluorescent dye rhodamine B (RhB) and vinblastine, by PFOS were evaluated using 24 and 48h post-fertilization (hpf) embryos. Upon 90min exposure of 24hpf embryos to 1µM RhB and different PFOS concentrations (3-300µM) accumulation of RhB in zebrafish was increased by up to 11.9-fold compared to controls, whereas RhB increases in verapamil treatments were 1.7-fold. Co-administration of PFOS and vinblastine in exposures from 0 to 48hpf resulted in higher vinblastine-caused mortalities in zebrafish embryos indicating increased uptake of this compound. Interference of PFOS with zebrafish Abcb4 activity was further studied using recombinant protein obtained with the baculovirus expression system. PFOS lead to a concentration-dependent decrease of the verapamil-stimulated Abcb4 ATPase activity; at higher PFOS concentrations (250, 500µM), also the basal ATPase activity was lowered indicating PFOS to be an Abcb4 inhibitor. In exposures of 48hpf embryos to a very high RhB concentration (200µM), accumulation of RhB in embryo tissue and adsorption to the chorion were increased in the presence of 50 or 100µM PFOS. In conclusion, the results indicate that PFOS acts as inhibitor of zebrafish Abcb4; however, the exceptionally large PFOS-caused effect amplitude of RhB accumulation in the 1µM RhB experiments and the clear PFOS effects in the experiments with 200µM RhB suggest that an additional mechanism appears to be responsible for the potential of PFOS to enhance uptake of Abcb4 substrates.

Endocrine disruption of water and sediment extracts in a non-radioactive dot blot/RNAse protection-assay using isolated hepatocytes of rainbow trout.

GOAL, SCOPE AND BACKGROUND: In order to evaluate the estrogenic activity of sediments and XAD water extracts of selected sites of the catchment area of the River Neckar, a river system in Southern Germany, an integrative assessment approach was used to assess the ecological hazard potential of endocrine-disrupting compounds in sediment and water. METHODS: The approach is based on estrogen receptor-mediated vitellogenin synthesis induced in isolated hepatocytes of rainbow trout and quantified in a non-radioactive dot blot/RNAse protection-assay in parallel to comprehensive chemical analyses of estrogenic substances. RESULTS AND DISCUSSION: Numerous investigated extracts revealed an estrogen activity comparable to that of the positive control (1 nM 17beta-estradiol corresponding to 270 ng/L in the test medium). Based on a concentration factor of 30 in the extracts and a recovery of XAD resins of approximately 80%, 17beta-estradiol equivalent concentrations between 20 and 26.7 ng/L could be calculated downstream of a sewage treatment plant (< 0.1 ng/L for a reference site). A comparison of the bioassay-derived Bio-TEQs (toxicity equivalents) and the Chem-TEQs revealed a high correlation with a Pearson coefficient of 0.85, indicating that the same ranking of the samples could be obtained with respect to the endocrine disrupting potential with both chemical and bioanalytical analysis. However, the TEQ concentrations computed from chemical analyses were significantly lower than the bioassay-derived TEQ concentrations. In fact, in none of the samples, more than 14% of the vitellogenin-inducing potency could be attributed to the substances (steroids, alkylphenols, bisphenol A, diethylstilbestrol) analyzed. A comparison of the endocrine disrupting potential of sediments extracted by the solvents acetone and methanol revealed lower biological effects for acetone-extracted samples. Possible reasons may be a masking of endocrine effects in acetone extracts by cytotoxicity, a low extraction efficiency of the solvent acetone, or anti- estrogen potencies of some extracted sediment compounds. Using a mass balance approach, the contribution of the compounds analyzed chemically (Chem-TEQs) to the total endocrine activity (Bio-TEQs) was calculated. Based on the very low detection limits, particularly of the steroids with their high TEF factors, results revealed that a calculation of the Chem-TEQs is associated with considerable scale inaccuracy: Whereas only 7-15% of the biological effectiveness (Bio-TEQs) could be explained by endocrine substances identified above the detection limits, the assumption of concentrations slightly below the given detection limits would result in a significant over estimation (137-197%) of the Bio-TEQs. Even the interassay variation of the dot blot assay with different fish donors for primary hepatocyte (factor 2-2.5) is relatively low, when compared to the large range of the Chem-TEQ concentrations (factor 20) obtained when applying different modes of calculation. CONCLUSIONS AND OUTLOOK: Overall, only a minor portion of the endocrine activity detected by bioassays could be linked to compounds identified by chemical analysis. In vitro assays for assessment of endocrine activities are useful as sensitive integrating methods that provide quantitative estimates of the total activity of particular receptor-mediated responses. Although discrepancies may also result from different bioanalytical approaches, it is overall likely that bioanalytical and not chemical analytical approaches give the correct estimate of endocrine disrupting potencies in environmental samples. As a conclusion, assessment of endocrine disruption based on chemical analysis alone does not appear sufficient and further research into the spectrum of substances with potential endocrine activity as well as into additive or even synergistic effects in complex environmental samples is urgently needed.

Accidental contamination of a German town's drinking water with sodium hydroxide.

Case report of a very serious drinking water incident putting up to 50,000 inhabitants of a town near Bonn in North Rhine-Westphalia, Germany at risk. A concentrated solution of highly alkaline water by sodium hydroxide was accidentally washed into the town's drinking water at a pumping station and increased the pH-value of the water to 12. Residents who came into contact with the contaminated water immediately had a toxic reaction. The incident was detected by complaints from customers and after that was stopped within several hours. The pipes were flushed and the customers were warned not to use the water till the all clear. After this immediate management there was an investigation and the cause of the incident was detected as an accidental release of accumulated sodium hydroxide (NaOH) solution. The lack of a network alarm system and the automatic cut-off mechanisms as deficiencies in the design of the station were rectified by the water company immediately after the incident.

Construction of an expression system for site-directed mutagenesis of the lantibiotic mersacidin.

The lantibiotic (i.e., lanthionine-containing antibiotic) mersacidin is an antimicrobial peptide of 20 amino acids which is produced by Bacillus sp. strain HIL Y-85,54728. Mersacidin inhibits bacterial cell wall biosynthesis by binding to the precursor molecule lipid II. The structural gene of mersacidin (mrsA) and the genes for the enzymes of the biosynthesis pathway, dedicated transporters, producer self-protection proteins, and regulatory factors are organized in a biosynthetic gene cluster. For site-directed mutagenesis of lantibiotics, the engineered genes must be expressed in an expression system that contains all of the factors necessary for biosynthesis, export, and producer self-protection. In order to express engineered mersacidin peptides, a system in which the engineered gene replaces the wild-type gene on the chromosome was constructed. To test the expression system, three mutants were constructed. In S16I mersacidin, the didehydroalanine residue (Dha) at position 16 was replaced with the Ile residue found in the closely related lantibiotic actagardine. S16I mersacidin was produced only in small amounts. The purified peptide had markedly reduced antimicrobial activity, indicating an essential role for Dha16 in biosynthesis and biological activity of mersacidin. Similarly, Glu17, which is thought to be an essential structure in mersacidin, was exchanged for alanine. E17A mersacidin was obtained in good yields but also showed markedly reduced activity, thus confirming the importance of the carboxylic acid function at position 17 in the biological activity of mersacidin. Finally, the exchange of an aromatic for an aliphatic hydrophobic residue at position 3 resulted in the mutant peptide F3L mersacidin; this peptide showed only moderately reduced activity.

Occurrence of toxigenic Aspergillus versicolor isolates and sterigmatocystin in carpet dust from damp indoor environments.

Over the past decade, there has been growing concern regarding the role of toxigenic fungi in damp indoor environments; however, there is still a lack of field investigations on exposure to mycotoxins. The goal of our pilot study was to quantify the proportion of toxigenic Aspergillus versicolor isolates in native carpet dust from damp dwellings with mold problems and to determine whether sterigmatocystin can be detected in this matrix. Carpet dust samples (n = 11) contained from <2.5 x 10(1) to 3.6 x 10(5) (median, 3.1 x 10(4)) A. versicolor CFU/g of dust, and the median proportion of A. versicolor from total culturable fungi was 18%. Based on thin-layer chromatography detection of sterigmatocystin, 49 of 50 A. versicolor isolates (98%) were found to be toxigenic in vitro. By using high-performance liquid chromatography-electrospray ionization tandem mass spectrometry, sterigmatocystin could be detected in low concentrations (2 to 4 ng/g of dust) in 2 of 11 native carpet dust samples. From this preliminary study, we conclude that most strains of A. versicolor isolated from carpet dust are able to produce sterigmatocystin in vitro and that sterigmatocystin may occasionally occur in carpet dust from damp indoor environments. Further research and systematic field investigation are needed to confirm our results and to provide an understanding of the health implications of mycotoxins in indoor environments.