Cadmium in soil, crops and resultant dietary exposure
Rietra, R.P.J.J. ; Mol, G. ; Rietjens, I.M.C.M. ; Römkens, P.F.A.M. - \ 2017
Wageningen : Wageningen Environmental Research (Wageningen Environmental Research rapport 2784) - 39
cadmium - soil - food intake - crops - exposure - fertilizers - food safety - toxicology - cadmium - bodem - voedselopname - gewassen - blootstelling - kunstmeststoffen - voedselveiligheid - toxicologie
TBT-gehalten en effecten bij de Gewone Alikruik, de Gevlochten Fuikhoorn en de Purperslak langs de Nederlandse kust in 2016
Kotterman, M. ; Jol, J. ; Barneveld, E. van - \ 2016
IJmuiden : Wageningen Marine Research (Wageningen Marine Research rapport C110/16) - 35
gastropoda - organo-tinverbindingen - bemonsteren - kustgebieden - toxicologie - nederland - gastropoda - organotin compounds - sampling - coastal areas - toxicology - netherlands
Aanpassing programma monitoring aal ter ondersteuning beleidskader open/gesloten gebieden
Kotterman, Michael - \ 2016
IJmuiden : Wageningen Marine Research (Wageningen Marine Research rapport C084/16) - 34
palingen - monitoring - visserij - toxicologie - visserijbeleid - eels - monitoring - fisheries - toxicology - fishery policy
Dit rapport beschrijft een aanpassing van het bestaande aalmonitoringsprogramma dat ingezet kan worden voor het te ontwikkelen beleidskader. De aanpassingen zijn op basis van bestaande data. Dit nieuwe protocol is voor het eerst toegepast op het aal monitoringsprogramma in 2016.
Overdracht van contaminanten van moeder naar jong en chemische profielen in bruinvissen gestrand langs de Nederlandse kust
Heuvel-Greve, M.J. van den; Kwadijk, C.J.A.F. ; Kotterman, M.J.J. - \ 2016
Wageningen Marine Research (Rapport / Wageningen Marine Research C096/16) - 30
phocoena - besmetters - overdracht - pasgeboren dieren - polychloorbifenylen - toxische stoffen - toxicologie - phocoena - contaminants - transfer - newborn animals - polychlorinated biphenyls - toxic substances - toxicology
Nederland heeft ten aanzien van de bruinvis een beschermingsplicht onder de Natuurbeschermingswet 1998. Om deze plicht goed te kunnen invullen is in 2011 het Bruinvisbeschermingsplan opgesteld. Dit beschermingsplan laat een aantal kennisleemten zien, o.a. ten aanzien van de rol die verontreinigingen spelen in de sterfte onder bruinvissen. In de afgelopen decennia is het aantal bruinvissen dat aanspoelt langs de Nederlandse kust toegenomen. Om te bepalen of aanwezigheid en gehalten van contaminanten in aangespoelde bruinvissen mogelijke effecten hebben gehad op hun gezondheid, zijn een aantal specifieke kennisvragen opgesteld die in dit rapport worden beantwoord. Er is in dit onderzoek gebruik gemaakt van monsters van aangespoelde bruinvissen gevonden in zowel Nederland als Denemarken.
Persistent organic pollutants : aberrant DNA methylation underlying potential health effects
Dungen, M.W. van den - \ 2016
Wageningen University. Promotor(en): Tinka Murk; Ellen Kampman, co-promotor(en): Wilma Steegenga; Dieuwertje van Gils-Kok. - Wageningen : Wageningen University - ISBN 9789462577893 - 207
persistent organic pollutants - dna methylation - molecular genetics - epigenetics - health hazards - toxic substances - endocrine disruptors - eels - fish consumption - toxicology - persistente organische verontreinigende stoffen - dna-methylering - moleculaire genetica - epigenetica - gezondheidsgevaren - toxische stoffen - hormoonverstoorders - palingen - visconsumptie - toxicologie
Wild caught fish, especially marine fish, can contain high levels of persistent organic pollutants (POPs). In the Netherlands, especially eel from the main rivers have high POP levels. This led to a ban in 2011 on eel fishing due to health concerns. Many of the marine POPs have been related to adverse health effects such as endocrine disruption, neurodevelopmental problems, immune suppression and cancer. Although some mechanisms of action of POPs are clear, like dioxins binding to the aryl hydrocarbon receptor and OH-PCBs binding to thyroid transport proteins, not all adverse health effects can be explained by these mechanisms of action. Epigenetic phenomena, such as DNA methylation, have been proposed as a possible molecular mechanism underlying adverse health effects. DNA methylation is a heritable modification, which refers to the addition of a methyl group to cytosine in a CpG dinucleotide. Observational studies have indeed shown that POPs can affect global DNA methylation, although results are inconsistent. Some animal studies as well as in vitro experiments suggest that POPs can affect gene-specific DNA methylation, however, the biological significance and relevance for humans is not clear. Therefore, this thesis aimed to 1) study the accumulation of POPs in men consuming eel from high-polluted areas 2) elucidate whether seafood-related POPs can induce aberrant DNA methylation and 3) to determine whether DNA methylation is related to functional endpoints and gene expression in vitro.
For this purpose eight POPs that are abundantly present in seafood were chosen, namely 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), polychlorobiphenyl (PCB) 126 and 153, perfluorooctanesulfonic acid (PFOS), hexabromocyclododecane (HBCD), 2,2′,4,4′- tetrabromodiphenyl ether (BDE-47), tributyltin (TBT), and methylmercury (MeHg). Chapter 2 describes the in vitro effects of these POPs and mixtures thereof in H295R adrenocortical carcinoma cells. Relative responses for 13 steroid hormones and 7 genes involved in the steroidogenic pathway, and CYP1A1, were analysed. PFOS induced the most pronounced effects on steroid hormone levels by significantly affecting 9 out of 13 hormone levels measured, with the largest increases found for 17β-estradiol, corticosterone, and cortisol. Furthermore, TCDD, both PCBs, and TBT significantly altered steroidogenesis. Increased steroid hormone levels were accompanied by related increased gene expression levels. The differently expressed genes were MC2R, CYP11B1, CYP11B2, and CYP19A1 and changes in gene expression levels were more sensitive than changes in hormone levels. The POP mixtures tested showed mostly additive effects, especially for DHEA and 17β-estradiol levels. This study shows that some seafood POPs are capable of altering steroidogenesis in H295R cells at concentrations that mixtures might reach in human blood, suggesting that adverse health effects cannot be excluded. DNA methylation was not measured in this study due to the short exposure time, which was expected not to be sufficient for long-term epigenetic marks. Therefore, in chapters 3A and 3B a differentiation experiment was performed enabling long-term exposure to POPs. Human mesenchymal stem cells (hMSCs) were differentiated into mature adipocytes over a time-course of 10 days. The transcriptional regulatory cascade involved in adipocyte differentiation has been extensively studied, however the mechanisms driving the transcription are poorly understood. In chapter 3A we therefore first explored the involvement of DNA methylation in transcriptional regulation during adipocyte differentiation. Genome-wide changes in DNA methylation were measured as well as the expression of adipogenic genes. The majority of these genes showed significant expression changes during the differentiation process. There were, however, only a couple of these differentially expressed genes that were differentially methylated. Genome-wide DNA methylation changes were most often located in intergenic regions, and underrepresented close to the transcription start site. This suggested that changes in DNA methylation are not the underlying mechanism regulating gene expression during adipocyte differentiation. Nevertheless, we explored DNA methylation differences after continuous exposure to POPs to investigate whether this could be an underlying mechanism by which POPs affect adipocyte differentiation. TCDD and PFOS decreased lipid accumulation, while TBT increased lipid accumulation. TCDD and TBT induced opposite gene expression profiles, whereas after PFOS exposure gene expression remained relatively stable. Genome-wide DNA methylation analysis showed that all three POPs affected DNA methylation patterns in adipogenic and other genes, but without concomitant gene expression changes. Differential methylation was again predominantly detected in intergenic regions, where the biological relevance of alterations in DNA methylation is unclear. This study demonstrated that POPs, at environmentally relevant levels, are able to induce differential DNA methylation in differentiating adipocytes. However, the biological relevance of this aberrant DNA methylation remains unclear.
The in vitro results showed a proof of principle that POPs could be capable of altering DNA methylation. To this date, no human studies were performed investigating the relationship between POP levels and genome-wide DNA methylation. In order to investigate this, we first measured POP levels in eel consumers from the high-polluted areas (areas with a ban on eel fishing) and compared these levels to men consuming eel from low-polluted areas or aquaculture (chapter 4). We aimed to investigate the accumulation of these POPs and determine whether the predictions made in an earlier risk assessment were valid. This was indeed the case as levels of dioxins and dioxin-like compounds were on average 2.5 times higher in men consuming eel from high-polluted areas. Furthermore, PCBs with their hydroxylated metabolites, and perfluoroalkyl substances (PFASs) were, up to ten times, higher in these consumers. Especially the high levels of dioxins and dioxin-like compounds as well as the OH-PCBs are expected to be of health concern. We continued this research in chapter 5 by associating all the measured POPs to clinical parameters related to e.g. thyroid hormones and liver enzymes, but found no relationship. Subsequently, we investigated the association between dioxins and dioxin-like compounds, the sum of seven indicator PCBs, and PFOS with genome-wide DNA methylation. We detected a number of differentially methylated regions (DMRs) related to genes involved in carcinogenesis (e.g. BRCA1, MAGEE2, HOXA5), the immune system (e.g. RNF39, HLA-DQB1), in retinol homeostasis (DHRS4L2), or in metabolism (CYP1A1). In contrast to the in vitro data, most significant effects were detected in CpG islands and were annotated close to the promoter region. This suggests that the differential methylation might be related to differential expression and possibly induce adverse health effects. The hypermethylation of some of these gene related to cancer could be an explanation of the carcinogenic effects that are observed with POP exposure.
Based on the results of this thesis we can conclude that the consumption of eel from high-polluted areas lead to accumulation of POPs above safe levels and that POP levels are associated with gene-specific DNA methylation in vitro as well as in environmentally exposed men. More research, however, is needed to fully elucidate the biological implications of this aberrant DNA methylation. A first step can be to measure histone modifications, as these two epigenetic marks together are likely better in predicting gene expression. The second step can be to investigate the potential health effects related to these epigenetic marks and to determine whether there is a causal relationship. Although at this point there is a lack of knowledge with regard to health effects caused by DNA methylation, the consumption of eel from these high-polluted areas is ill- advised, because adverse health effects cannot be excluded based on our results and can even be expected based on literature.
Proteomics as a tool to gain more insight into sub-lethal toxicological effects
Miller, Ingrid - \ 2016
Wageningen University. Promotor(en): Tinka Murk, co-promotor(en): A.C. Gutleb; T. Serchi. - Wageningen : Wageningen University - ISBN 9789462578210 - 182
proteomics - laboratory methods - sublethal effects - toxic substances - endocrine disruptors - food consumption - toxicology - animal experiments - eiwitexpressieanalyse - laboratoriummethoden - subletale effecten - toxische stoffen - hormoonverstoorders - voedselconsumptie - toxicologie - dierproeven
This thesis focuses on a modern analytical method, proteomics, to investigate its use in the field of toxicological research. Proteomics is a high resolution method which separates all proteins present in a sample at a clearly defined state and compares this pattern to another one, under slightly different conditions (e.g. after exposure to a chemical). Protein changes may give rise to or reflect disease/harm of the individual and can be attributed to alterations in body functions/regulation systems. Analysis conditions and different varieties of proteomic methods are explained, and a brief introduction given where proteomics is already applied in toxicology. A specific investigation has been performed with the flame retardant HBCD (i.e. hexabromocyclododecane). It is a compound that accumulates in lipid tissue from where it is only slowly removed. Its mechanism of action is not yet completely understood and sometimes seems to be contradictory. Rats were exposed to HBCD in very low doses for just one week and liver proteins were compared to those of unexposed animals. As HBCD is suggested to disturb the thyroid system, both healthy and hypothyroid rats were investigated, of both genders. In female rats, not in males, some specific liver protein changes were seen in glucose/carbohydrate and lipid metabolism, and also in some stress related proteins. Changes were not dependent on the thyroid function of the females. These results are in line with previous findings that female rats were more susceptible to HBCD than males. In a further step, protein patterns of unexposed animals of both genders were compared, revealing gender-dependent differences that exceeded the effects seen in any of the other comparisons, mainly in the pathways that were also affected by HBCD in females. A previous proteomic study on serum proteins has also shown clear gender-dependent concentration differences in rats. This underlines the importance of performing studies both in female and male individuals. The detection of considerable gender-dependent protein alterations confirms that proteomics is a biochemical tool with high sensitivity and large potential also in toxicological research.
Dermal absorption and toxicological risk assessment : pitfalls and promises
Buist, H. - \ 2016
Wageningen University. Promotor(en): Ruud Woutersen; Ivonne Rietjens, co-promotor(en): J.J.M. van de Sandt. - Wageningen : Wageningen University - ISBN 9789462577275 - 200
skin - absorption - permeability - in vitro - experiments - exposure assessment - risk assessment - toxicology - biocides - rodenticides - preservatives - disinfection - huid - absorptie - permeabiliteit - in vitro - experimenten - blootstellingsbepaling - risicoschatting - toxicologie - biociden - rodenticiden - conserveermiddelen - desinfectie
Absorption of toxic substances via the skin is an important phenomenon in the assessment of the risk of exposure to these substances. People are exposed to a variety of substances and products via the skin, either directly or indirectly, while at work, at home or in public space. Pesticides, organic solvents and metalworking fluids are seen to be important contributors to adverse health effects due to occupational exposure via the skin. In daily life, cosmetics, clothing and household products are the most relevant commodities with respect to exposure via the skin.
Given the importance of skin exposure in the assessment of the risk of toxic substances, the objective of this thesis was to further develop, evaluate and improve methods for including skin absorption data this assessment.
In this thesis, four factors influencing dermal absorption, namely dermal loading (chapters 3 and 6), irritative/corrosive potential (chapters 3 and 4), frequency of exposure (chapters 3, 4 and 5) and the vehicle used (chapter 5), were investigated in more detail. Furthermore, a model to extrapolate infinite dose absorption data to finite dose conditions, baptized Dermal Absorption Model for Extrapolation (DAME), was developed and tested.
n chapter 2 of this thesis, the relationship between relative dermal absorption and dermal loading was investigated. Hundred-and-thirty-eight dermal publicly available absorption experiments with 98 substances were evaluated. The results obtained revealed that dermal loading ranged mostly between 0.001 and 10 mg/cm2. In 87 experiments (63%), an inverse relationship was observed between relative dermal absorption and dermal loading. On average, relative absorption at high dermal loading was 33 times lower than at low dermal loading. Known skin irritating and volatile substances less frequently showed an inverse relationship between dermal loading and relative absorption. It was concluded that when using relative dermal absorption in regulatory risk assessment, its value should be determined at or extrapolated to dermal loadings relevant for the exposure conditions being evaluated.
n chapter 3 of this thesis, a literature search was presented with the aim to investigate whether neglecting the effects of repeated exposure may lead to an incorrect estimate of dermal absorption. The results demonstrated that the effect of repeated versus single exposure does not demonstrate a unique trend. Nevertheless, an increase in daily absorption was frequently observed upon repeated daily exposure. The little information available mostly concerned pharmaceuticals. However, consumers and workers may be repeatedly exposed to other types of chemicals, like disinfectants and cleaning products, which often contain biocidal active substances that may decrease the barrier function of the skin, especially after repeated exposure. These biocidal products, therefore, may present a safety risk that is not covered by the current risk assessment practice since absorption data are usually obtained by single exposure experiments. Consequently, it was decided to investigate the importance of this issue for biocide safety evaluation. As the literature search revealed that hardly any data on absorption upon repeated dermal exposure to biocides are available, it was concluded that data need to be generated by testing.
To cover the entire range of biocidal products in such testing, a representative series of biocidal substances should be tested, making in vitro testing of dermal absorption the preferred choice over in vivo testing. Based on an inventory made, it appeared that the 16 product types represented among the biocidal products authorised in the Netherlands could be clustered into 6 more or less homogeneous categories based on similarity in active substances. This result could facilitate experimental testing by providing a basis for selection of a limited number of representative compounds to be evaluated.
n chapter 4 of this thesis, the importance of the effect of repeated dermal exposure on skin permeability for biocide safety evaluation was investigated, using a selection of nine representative biocides from the inventory made in chapter 3. The in vitro dermal penetration of tritiated water and [14C]propoxur was chosen as a measure of the permeability and integrity of human abdominal skin after single and repeated exposure. The results indicated that single and repeated exposure to specific biocidal products (e.g. the quaternary ammonium chlorides DDAC and ADBAC) may significantly increase skin permeability, especially when the compounds are applied at high concentrations, while a substance like formaldehyde may reduce skin permeability under specific conditions.
n chapter 5 of this thesis, the in vitro dermal absorption kinetics of the quaternary ammonium compound didecyldimethylammonium chloride (DDAC) during single and repeated exposure was studied in more detail. In addition, the influence of biocidal formulations on the absorption of DDAC was investigated, because it was expected that formulation characteristics may be another factor influencing its dermal absorption. The analysis of biocidal products on the Dutch market, reported in chapter 3, indicated that DDAC is often used in combination with other active ingredients. DDAC was most frequently combined with formaldehyde, glutaraldehyde and/or alkyldimethylbenzylammonium chloride (ADBAC). Consequently, commercial formulations containing one or more of these additional active ingredients were selected, in addition to one formulation containing only DDAC as an active ingredient. The selected commercial formulations tended to reduce skin penetration of DDAC. This was most pronounced with the formulation containing the highest concentration of formaldehyde (196 mg/mL) and glutaraldehyde (106 mg/mL), which reduced the flux of DDAC across the skin by 95%. The reduction caused by the only tested formulation containing no other active ingredients than DDAC, and thus incorporating no aldehydes, was smallest, and did not reach statistical significance.
n chapter 6 of this thesis, a simple in silico model to predict finite dose dermal absorption from infinite dose data (kp and lag time) and the stratum corneum/water partition coefficient (KSC,W) was developed. This model was tentatively called Dermal Absorption Model for Extrapolation (DAME). As dermal exposure may occur under a large variety of conditions leading to quite different rates of absorption, such a predictive model using simple experimental or physicochemical inputs provides a cost-effective means to estimate dermal absorption under different conditions.
To evaluate the DAME, a series of in vitro dermal absorption experiments was performed under both infinite and finite dose conditions using a variety of different substances. The kp’s and lag times determined in the infinite dose experiments were entered into DAME to predict relative dermal absorption value under finite dose conditions. For six substances, the predicted relative dermal absorption under finite dose conditions was not statistically different from the measured value. For all other substances, measured absorption was overpredicted by DAME, but most of the overpredicted values were still lower than 100%, the European default absorption value for the tested compounds.
In conclusion, our finite dose prediction model (DAME) provides a useful and cost-effective estimate of in vitro dermal absorption, to be used in risk assessment for non-volatile substances dissolved in water at non-irritating concentrations.
n chapter 7 of this thesis, the results of the research reported in chapters 2 to 6 were put into perspective, the pitfalls and promises emanating from them discussed and general conclusions drawn. The possible influence of vehicles on absorption and the possible impact of irritative or corrosive vehicles or chemicals on the skin barrier have been demonstrated in this thesis. An in silico predictive model tentatively called DAME was developed, which enables the user to evaluate a variety of dermal exposure scenarios with limited experimental data (kp and lag time) and easy to obtain physicochemical properties (MW and log KOW). The predictions of our experiments reported in chapter 6 were compared to those of the Finite Dose Skin Permeation (FDSP) model published on the internet by the US Centers for Disease Control and Prevention (CDC). DAME outperformed FDSP (R2 of the correlation predicted/measured potential absorption 0.64 and 0.12, respectively). At present, the applicability domain of DAME is limited to non-volatile substances dissolved in aqueous solvents. However, in future the model will be adapted to include volatile substances as well.
Altogether, it is concluded that dermal exposure can be an important factor in risks posed by chemicals and should be taken into account in risk assessment. The methods to actually do this are still open for further improvement to better account for the various factors influencing skin penetration and to develop adequate combinations of in vitro and in silico models that can accurately predict human dermal absorption.
Replacing animal experiments in developmental toxicity testing of phenols by combining in vitro assays with physiologically based kinetic (PBK) modelling
Strikwold, Marije - \ 2016
Wageningen University. Promotor(en): Ivonne Rietjens; Ruud Woutersen, co-promotor(en): Ans Punt. - Wageningen : Wageningen University - ISBN 9789462576926 - 169
animal experiments - animal testing alternatives - toxicity - testing - phenols - in vitro - embryonic stem cells - tissues - cells - dosage - toxicology - animal health - dierproeven - alternatieven voor dierproeven - toxiciteit - testen - fenolen - in vitro - embryonale stamcellen - weefsels - cellen - dosering - toxicologie - diergezondheid
Estimation of degradation rates in cosm water : Guidance for inverse modelling using TOXSWA
Deneer, J.W. ; Adriaanse, P.I. ; Griethuysen, C. van; Boesten, J.J.T.I. - \ 2015
Wageningen : Alterra, Wageningen-UR (Alterra-rapport 2679) - 151
pesticides - degradation - water - models - aquatic toxicology - toxicology - testing - pesticiden - degradatie - water - modellen - aquatische toxicologie - toxicologie - testen
Physiologically based in silico modelling to examine DNA adduct formation by different food-borne a,ß-unsaturated aldehydes at realistic low dietary exposure levels
Kiwamoto, R. - \ 2015
Wageningen University. Promotor(en): Ivonne Rietjens, co-promotor(en): Ans Punt. - Wageningen : Wageningen University - ISBN 9789462572843 - 200
aldehyden - dna - ontgifting - voedseladditieven - aromatische stoffen - genotoxiciteit - carcinogenen - modellen - wiskundige modellen - fysiologie - simulatiemodellen - toxicologie - aldehydes - dna - detoxification - food additives - flavourings - genotoxicity - carcinogens - models - mathematical models - physiology - simulation models - toxicology
Abstract (R.Kiwamoto ISBN 978-94-6257-284-3)
Various α,β-unsaturated aldehydes are present in fruits, vegetables, spices, or processed products containing these items as natural constituents or as added food flavouring agents. Because of the α,β-unsaturated aldehyde moiety the β carbon in the molecule becomes electron deficient and the aldehydes react with electron rich molecules including DNA via Michael addition. The formation of DNA adducts raises a concern for genotoxicity, although formation of DNA adducts may not be significant at low doses relevant for dietary exposure in vivo because of adequate detoxification. This thesis therefore aimed at determining dose-dependent detoxification and DNA adduct formation of food-borne α,β-unsaturated aldehydes by using a physiologically based in silico modelling approach in order to contribute to the safety assessment of these aldehydes used as food flavourings instead of performing animal experiments.
Physiologically based in silico models were developed for 18 α,β-unsaturated aldehydes. The model outcomes indicated that the DNA adduct formation by the 18 α,β-unsaturated aldehydes as food flavourings is negligible and does not raise a safety concern at their levels of intake resulting from their use as food flavourings. The application of QSAR models strongly accelerated the development process of the PBK/D models of the group of 18 compounds. Also, it was illustrated that physiologically based in silico models provide a very useful and powerful tool to facilitate a group evaluation and read-across for food-borne DNA reactive agents. PBK/D models developed for the group of compounds supported read-across from cinnamaldehyde which is known not to be genotoxic or carcinogenic in vivo to other aldehydes, by allowing comparison of dose-dependent DNA adduct formations. Altogether this thesis presented physiologically based in silico modelling as an approach to test relevance of positive in vitro genotoxicity results by DNA reactive compounds in vivo without using animal experiments.
Sediment properties in five Dutch watercourses : indicative measurements for the registration procedure of plant protection products in The Netherlands
Adriaanse, P.I. ; Crum, S.J.H. ; Elbers, J.A. ; Massop, H.T.L. ; Beltman, W.H.J. - \ 2015
Wageningen : Alterra (Alterra report 2574) - 93
sloten - waterbodems - toxicologie - pesticiden - akkerbouw - tuinbouw - oppervlaktewater - ditches - water bottoms - toxicology - pesticides - arable farming - horticulture - surface water
Van vijf sloten verspreid liggend over Nederland, zijn de sediment eigenschappen droge bulkdichtheid, organische-stofgehalte en porositeit zijn als functie van de diepte gemeten. Het betreft hier akkerbouw- en tuinbouwgebieden. De gevonden waarden verschillen duidelijk van de gebruikte waarden voor sediment in de oppervlaktewater scenario’s, die momenteel worden ontwikkeld voor de Nederlandse registratieprocedure voor gewasbeschermingsmiddelen (die zijn overgenomen van de EU-FOCUS scenario’s).
Use of the ES-D3 cell differentiation assay, combined with the BeWo transport model, to predict relative in vivo developmenatl toxicity of antifungal compounds
Li, H. ; Rietjens, I. ; Louisse, J. ; Blok, M. ; Wang, X. ; Snijders, L. ; Ravenzwaay, B. van - \ 2015
Toxicology in Vitro 29 (2015)2. - ISSN 0887-2333 - p. 320 - 328.
dose-response curves - placental perfusion - test est - vitro - toxicology - potency - risk - rat
We investigated the applicability of the ES-D3 cell differentiation assay combined with the in vitro BeWo transport model to predict the relative in vivo developmental toxicity potencies. To this purpose, the in vitro developmental toxicity of five antifungal compounds was investigated by characterizing their inhibitory effect on the differentiation of ES-D3 cells into cardiomyocytes. The BeWo transport model, consisting of BeWo b30 cells grown on transwell inserts and mimicking the placental barrier, was used to determine the relative placental transport velocity. The ES-D3 cell differentiation data were first compared to benchmark doses (BMDs) for in vivo developmental toxicity as derived from data reported in the literature. Correlation between the benchmark concentration for 50% effect (BMCd50) values, obtained in the ES-D3 cell differentiation assay, with in vivo BMD10 values showed a reasonable correlation (R2 = 0.57). When the ES-D3 cell differentiation data were combined with the relative transport rates obtained from the BeWo model, the correlation with the in vivo data increased (R2 = 0.95). In conclusion, we show that the ES-D3 cell differentiation assay is able to better predict the in vivo developmental toxicity ranking of antifungal compounds when combined with the BeWo transport model, than as a stand-alone assay.
A Tutorial for Analysing the Cost-effectiveness of Alternative Methods for Assessing Chemical Toxicology: The Case of Acute Oral Toxicity Prediction
Norlen, H. ; Worth, A.P. ; Gabbert, S.G.M. - \ 2014
ATLA-Alternatives To Laboratory Animals 42 (2014)2. - ISSN 0261-1929 - p. 115 - 127.
health - cytotoxicity - foundations - toxicology
Compared with traditional animal methods for toxicity testing, in vitro and in silico methods are widely considered to permit a more cost-effective assessment of chemicals. However, how to assess the cost-effectiveness of alternative methods has remained unclear. This paper offers a user-oriented tutorial for applying cost-effectiveness analysis (CEA) to alternative (non-animal) methods. The purpose is to illustrate how CEA facilitates the identification of the alternative method, or the combination of methods, that offers the highest information gain per unit of cost. We illustrate how information gains and costs of single methods and method combinations can be assessed. By using acute oral toxicity as an example, we apply CEA to a set of four in silico methods (ToxSuite, TOPKAT, TEST, ADMET Predictor), one in vitro method (the 3T3 Neutral Red Uptake cytotoxicity assay), and various combinations of these methods. Our results underline that in silico tools are more cost-effective than the in vitro test. Battery combinations of alternative methods, however, do not necessarily outperform single methods, because additional information gains from the battery are easily outweighed by additional costs
Screening van hot spots van nieuwe verontreinigingen : een pilot studie in bodem, grondwater en oppervlaktewater
Lahr, J. ; Laak, T.L. ter; Derksen, A. - \ 2014
Wageningen : Alterra (Alterra-rapport 2538) - 87
bodemverontreiniging - waterverontreiniging - verontreinigende stoffen - toxicologie - geneesmiddelen - ecologische risicoschatting - biotesten - inventarisaties - soil pollution - water pollution - pollutants - toxicology - drugs - ecological risk assessment - bioassays - inventories
Onder nieuwe verontreinigingen verstaan we stoffen die nog niet of niet volledig zijn gereguleerd en waarvan de milieurisico’s vaak onbekend zijn. Daarbij gaat het om stoffen als natuurlijke hormonen en hormoonverstorende stoffen (weekmakers, detergenten, brandvertragers, e.d.), humane geneesmiddelen, diergeneesmiddelen, nanodeeltjes en microplastics. In de ‘waterwereld’ is altijd meer aandacht besteed aan de nieuwe verontreinigingen dan binnen andere beleidsvelden. In de bodem zijn de aanwezigheid en de mogelijke risico’s grotendeels onbekend. In 2013 heeft een consortium van diverse onderzoeksinstanties en stakeholders een pilotonderzoek uitgevoerd naar de aanwezigheid en mogelijke risico’s van hormonen en geneesmiddelen in het systeem bodem - grondwater - oppervlaktewater.
FOCUS_TOXSWA manual 4.4.2 : User’s Guide version 4
Beltman, W.H.J. ; Horst, M.M.S. ter; Adriaanse, P.I. ; Jong, A. de; Deneer, J.W. - \ 2014
Wageningen : Wettelijke Onderzoekstaken Natuur & Milieu (WOt-technical report 14) - 130
toxicologie - waterverontreiniging - waterbodems - pesticiden - waterlopen - plassen - modellen - toxicology - water pollution - water bottoms - pesticides - streams - ponds - models
The FOCUS_TOXSWA model calculates exposure concentrations of pesticides and their metabolites in watercourses and ponds. These concentrations are used in the pesticide registration procedure at EU level. The model concepts of TOXSWA are described briefly. The input files, output files, and the use of the graphical user interface to access the input and output are described. Input data are stored in a database. Pesticide entries resulting from drainage or runoff/erosion are accessed from separate files generated by FOCUS_MACRO and FOCUS_PRZM. Substance properties are accessed from the SPIN tool/database. Instructions for simulating a water-sediment study and a multi-year run are given
The Isolated Chicken Eye test to replace the Draize test in rabbits : from development to implementation: “The long and winding road”
Prinsen, M.K. - \ 2014
Wageningen University. Promotor(en): Ruud Woutersen; C.F.M. Hendriksen, co-promotor(en): C.A.M. Krul. - Wageningen : Wageningen University - ISBN 9789462570030 - 184
ogen - konijnen - kippen - laboratoriumdieren - alternatieven voor dierproeven - dierproeven - toxiciteit - toxicologie - histopathologie - dierenwelzijn - eyes - rabbits - fowls - laboratory animals - animal testing alternatives - animal experiments - toxicity - toxicology - histopathology - animal welfare
Dit proefschrift beschrijft de ontwikkeling, optimalisatie, validatie (binnen TNO) en toepassing van een alternatieve test met geïsoleerde ogen van kippen (de Isolated Chicken Eye, kortweg de ICE test) en in bredere zin de internationale validatie en acceptatie van de ICE test door overheidsinstanties.
Onderzoek naar inheemse wilde fauna, verslag over 2013
Tulden, P.W. van - \ 2014
Lelystad : Central Veterinary Institute (Rapport / Central Veterinairy Institute 14/CVI0014) - 31
fauna - wild - vogels - vissen - zoogdieren - toxicologie - pathologie - doodsoorzaken - monitoring - fauna - wildlife - birds - fishes - mammals - toxicology - pathology - causes of death - monitoring
Met betrekking tot een aantal opgedragen Wettelijke Onderzoekstaken (WOT) treedt het Central Veterinairy Institute op als Nationaal Referentie Laboratorium (NRL) voor aangifteplichtige virale, parasitaire, protozoaire en bacteriële ziekten, TSE’s en antimicrobiële resistentie. Dit verslag geeft een overzicht van onderzoek aan de dieren, die opgestuurd zijn naar het CVT. Enkele cijfers: 137 inzendingen in het kader van het wettelijke wilde fauna onderzoek. Maar ook 188 kadavers, 6 levende watervogels en 27 monsters verdacht materiaal. De kadavers zijn te verdelen in 65 roofvogels, 70 watervogels, 13 overige vogels, 26 zoogdieren en 14 vissen. Hier kon van 115 van de 188 kadavers de doodsoorzaak worden achterhaald. De meest vastgestelde doodsoorzaken zijn trauma, vergiftiging, uitputting, afschot en botulisme, afhankelijk van de diercategorie.
Protein biomarker-based screening for detection of recombinant bovine somatotropin abuse in dairy cows
Ludwig, S.K.J. - \ 2014
Wageningen University. Promotor(en): Michel Nielen, co-promotor(en): Leen van Ginkel. - Wageningen : Wageningen University - ISBN 9789462570146 - 248
somatotropine - melkvee - besmetters - biomarkers - recombinant eiwitten - toxicologie - detectie - somatotropin - dairy cattle - contaminants - biomarkers - recombinant proteins - toxicology - detection
Recombinant bovine somatotropin (rbST) is a 22 kDa proteohormone, which can be used to increase milk production in dairy cows. It has been marketed since 1994 and while its use in food production is approved in several countries, such as the US, it is banned in the EU since 2000. To enforce the ban on rbST in the EU and to control for ‘rbST-free’ –labelling in the US, detection methods are required that identify whether rbST has been used. Existing rbST detection methods focus on the detection of rbST itself in bovine serum. The recombinant form of the hormone has one amino acid exchanged at the N-terminus of the protein. RbST can therefore be potentially discriminated from the endogenous bST by mass spectrometric methods. Other methods employ sandwich enzyme-linked immunosorbent assays (ELISAs) with antibodies having a higher affinity to rbST than bST. These methods, however mainly lack sensitivity, reproducibility or selectivity for rbST and are therefore not widely applied. Hence, no method has been implemented so far to monitor rbST abuse in dairy farming. Screening methods developed for veterinary drug residue control in the EU have to perform according to Commission Decision 2002/657/EC and have to identify at least 95 % of the treated animals.
An alternative approach for rbST abuse detection is the analysis of rbST-dependent biomarkers. A biomarker is defined as an indicator of normal physiological processes, pathogenic processes, or pharmacological responses to a therapeutic intervention. Therefore, the levels of rbST-dependent protein biomarkers are either up- or downregulated after administration of rbST and rbST-specific biomarker profiles can be used to detect its abuse. RbST exerts similar physiological actions in the cow’s body as the endogenous bST. Therefore, proteins involved in the regulatory circuit of bST have been chosen as candidate biomarkers, such as insulin-like growth factor-1 (IGF-1), IGF binding protein 2 (IGFBP2) and osteocalcin. Additionally to that, the administration of rbST induces anti-rbST antibodies in the cow’s body, which can be detected as biomarkers. This approach is according to the growth hormone (GH) abuse detection in sports doping control, where solely protein biomarker profiles are used to identify the abuse.
Chapter 2 introduces protein biomarkers and how biomarkers can be used in sports doping and veterinary control to detect the abuse of illegal substances. The advantages of using biomarkers are that the biological effect of a substance usually lasts longer than the substance itself can be detected and therewith, the window of detection is expanded. Moreover, since different substances exert similar effects on physiological machineries for growth or production enhancement, biomarker-based-detection methods have the potential to detect a whole class of substances. Furthermore, low-dose mixtures of different banned substances, which might escape from direct detection of each individual substance used, could be still detected by the combined effect they exert. In this chapter, protein biomarker-based detection strategies are discussed against generic challenges in biomarker discovery and method development.
Part I of the thesis concerns biomarker analysis in serum and plasma samples from cattle, which are analysed using laboratory-based equipment.A triplex flow cytometric immunoassay (FCIA), which combines the detection of three rbST-dependent biomarkers, viz. IGF-1, IGFBP2 and anti-rbST antibodies is demonstrated in Chapter 3. Serum samples from treated and untreated dairy cows from a single animal study were analysed using this triplex FCIA. Characteristic treatment-dependent responses for all three individual biomarkers were shown. These results were combined using the statistical model k-nearest neighbours (kNN). This model discriminated rbST-treated from untreated cows with a truepositive rate of 89.1 % and a true-negative rate of 97.7 %.
This triplex FCIA was further extended with the biomarker osteocalcin and the resulting fourplex FCIA was used for biomarker profiling in serum samples from rbST-treated and untreated cows from two independent rbST treatment studies. In Chapter 4, different data analysis approaches were tested with the aim to detect the highest possible number of true-positive samples. The statistical model kNN was used on all 11 possible biomarker combinations and the combination of the biomarkers osteocalcin and endogenously produced antibodies against rbST proved to be very reliable and correctly predicted 95 % of the samples of treated cows starting from the second rbST injection until the end of the treatment period and even thereafter. With the same biomarker combination, only 12 % of the samples of untreated animals appeared false-positive. This reliability meets the requirements of Commission Decision 2002/657/EC for screening methods in veterinary control.
It can be expected that rbST-dependent biomarkers also show a response upon other treatments. Therefore in Chapter 5, the fourplex FCIA for rbST abuse detection was applied to bovines treated with steroids, such as estradiol, dexamethasone and prednisolone. Each treatment resulted in a specific plasma biomarker profile for IGF-1, IGFBP2, osteocalcin and anti-rbST antibodies, which could be distinguished from the profile of untreated animals. Therefore, the fourplex biomarker FCIA is, apart from rbST, also capable of detecting treatment with other growth-promoting agents and clearly shows the potential of biomarker profiling as a screening method in veterinary control.
Part II of the thesis focusses on protein biomarker analysis in milk samples and the change from laboratory-based to on-site analysis. In Chapter 6, the detection of anti-rbST antibodies in raw milk samples was demonstrated, which discriminated rbST-treated from untreated cows with a 67 % true-positive and 94 % true-negative rate. The laboratory-based assay was also applied to simulated tank milk and pasteurized milk samples. Using milk as a sample matrix for detection has the advantages of non-invasive sampling, and for tank milk analysis at the farm only one milk sample is needed to screen the whole farm for rbST (ab)use.
As a next step in Chapter 7, this assay was translated to an on-site pre-screening platform including a cellphone. Using this on-site platform, samples can be tested at the point where they were taken. Only samples that are suspect are transported to a laboratory for further analysis. To this end, a cellphone-based fluorescence imaging platform for the detection of anti-rbST antibodies in milk extracts was developed, which is based on a microsphere fluorescence immunoassay. After performing the assay, the fluorescence is excited by UV LEDs embedded in a dedicated cellphone attachment and the emitted fluorescence light is imaged by the cellphone camera. The fluorescence micro-images were analysed using a custom-developed Android application running on the same cellphone and milk samples from rbST-treated and untreated cows were discriminated.
Also in milk samples, the simultaneous detection of several biomarkers is advantageous as they can increase the confidence of a positive finding. Therefore in Chapter 8, a protein microarray-based platform for multiple rbST biomarker detection on a cellphone is presented, which detects anti-rbST antibodies and IGF-1 in milk samples. The 48 microspots on the microarray were labelled with Quantum Dots depending on the biomarker levels in the sample. Quantum Dot fluorescence was detected by the cellphone camera and the same opto-mechanical attachment as in Chapter 7 and images were analysed by custom software. RbST-treated clearly showed a treatment-dependent biomarker profile in milk that could be discriminated from the profile of untreated cows.
Future research should focus on the simultaneous detection of different targets of interest in milk samples, such as hormones, allergens, antibiotics, contaminants and other substances, all at the same time using the microarray platform on the cellphone. Moreover, sample handling can be facilitated by the use of pre-fabricated microfluidic devices including all required assay reagents. With the work presented in this thesis, screening for rbST abuse in serum and milk becomes possible: in the laboratory and on-site. The future implementation of these testing platforms for rbST abuse detection is a major leap forward concerning the enforcement of the rbST ban in the EU and concerning the value of protein biomarker-based approaches in veterinary control.
Consumer and farmer safety evaluation of application of botanical pesticides in black pepper crop protection
Hernandez-Moreno, J. ; Soffers, A.E.M.F. ; Wiratno, ; Falke, H.E. ; Rietjens, I. ; Murk, A.J. - \ 2013
Food and Chemical Toxicology 56 (2013). - ISSN 0278-6915 - p. 483 - 490.
clove oil - rotenone - ingestion - insecticides - toxicology - calamus - rat
This study presents a consumer and farmer safety evaluation on the use of four botanical pesticides in pepper berry crop protection. The pesticides evaluated include preparations from clove, tuba root, sweet flag and pyrethrum. Their safety evaluation was based on their active ingredients being eugenol, rotenone, ß-asarone and pyrethrins, respectively. Botanical pesticides from Acorus calamus are of possible concern because of the genotoxic and carcinogenic ingredient ß-asarone although estimated margins of exposure (MOE) for consumers indicate a low priority for risk management. For the other three botanical pesticides the margin of safety (MOS) between established acute reference doses and/or acceptable daily intake values and intake estimates for the consumer, resulting from their use as a botanical pesticide are not of safety concern, with the exception for levels of rotenone upon use of tuba root extracts on stored berries. Used levels of clove and pyrethrum as botanical pesticides in pepper berry crop production is not of safety concern for consumers or farmers, whereas for use of tuba root and sweet flag some risk factors were defined requiring further evaluation and/or risk management. It seems prudent to look for alternatives for use of sweet flag extracts containing ß-asarone
Alternatieve systematiek voor de beoordeling van covergistingsmaterialen : 1. toetsing van contaminanten aangewezen door de Meststoffenwet
Ehlert, P.A.I. ; Schöll, L. van; Dijk, T.A. van - \ 2013
Wageningen : Wettelijke Onderzoekstaken Natuur & Milieu (WOt-werkdocument 358) - 88
afvalverwerking - organisch afval - co-vergisting - beoordeling - milieubeleid - toxicologie - verontreinigingen - protocollen - biobased economy - waste treatment - organic wastes - co-fermentation - assessment - environmental policy - toxicology - impurities - protocols - biobased economy
Om het proces van de toelating van afval- en reststoffen voor gebruik als covergistingsmateriaal te versnellen en vooral ook om de verantwoordelijkheid voor de toetsing meer bij het bedrijfsleven te leggen, heeft de Staatsecretaris van het Ministerie van Economische Zaken (EZ) besloten tot opname van een ‘Alternatieve systematiek’ voor de toetsing van stoffen in de regelgeving. Met deze systematiek kan het bedrijfsleven op basis van een beperkt aantal gegevens zelfstandig beoordelen of een afval- of reststof geschikt is om te worden gebruikt als covergistingsmateriaal. Het Ministerie van EZ heeft Alterra Wageningen UR en RIVM gevraagd om een alternatieve systematiek op te stellen voor de toetsing van afval- en reststoffen voor gebruik als covergistingsmateriaal, waarbij een beperkt aantal gegevens nodig is. Tevens is gevraagd om mogelijkheden voor rubricering van afval- en reststoffen onder de rubricering van de Europese regelgeving voor afvalstoffen (EURAL-codes) in de Meststoffenwet te onderzoeken. Het onderzoek heeft geleid tot twee deelrapporten. Dit WOt-werkdocument (1) behandelt contaminanten die door de Meststoffenwet zijn aangewezen en die voor kunnen komen in afval- en reststoffen die bestemd worden voor vergisting met dierlijke mest. Het tweede WOt-werkdocument (De Poorter et al., 2013) behandelt organische microverontreinigingen die niet door de Meststoffenwet zijn aangewezen