monitoramento de dioxinas em incinerador na holanda

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Biomonitoring of dioxins/dl-PCBs in the north of the Netherlands; eggs of
backyard chickens, cow and goat milk and soil as indicators of pollution
Article · January 2014
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Abel Arkenbout
NGO ToxicoWatch
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Biomonitoring of Dioxins/dl-PCBs in the north of the Netherlands; 
 eggs of backyard chickens, cow and goat milk and soil as indicators of pollution 
 
Arkenbout, Abel 
Toxicowatch Foundation, Grote Ossenmarkt 18, 8861 CP Harlingen, the Netherlands 
 
 
 
Introduction 
 
In the region of Harlingen in the north of the Netherlands people have recently been very concerned about 
adverse health effects related to possible emissions of dioxins and other POPs by a recently installed waste 
incinerator. To evaluate these effects the local authorities make use of a very limited monitoring program. Using 
eggs of backyard chickens, cow and goat milk and soil as indicators we however set up a more adequate 
monitoring program. Because the Dutch authorities do not routinely check the eggs from small farms (or 
households) – with less than 200 chickens per farm – the risk of pollution with dioxins and dl-PCBs is 
underestimated. After finding several egg samples that did not comply with the Dutch regulation IKB for the 
commercial use of eggs (in case the level exceeds 1.75 pg BEQ / g fat one is compelled to take further action) we 
started a more extended monitoring program. 
 
 
 
Materials and methods 
 
Harlingen is a small city (c. 16.000 inhabitants) with agricultural industry and moreover a relatively low-profiled 
industry that is mainly concentrated near the harbour and consists of shipyards, metal working, an oil-company, 
fishing industry, a plastic company and a waste incinerator. Eggs of backyard chickens were sampled and 
analyzed, initially at 13 and in a second stage at 18 different locations near Harlingen. The eggs have been 
analysed for dioxins (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) by DR CALUX

 
(BioDetection Systems, Amsterdam and RIKILT, Wageningen (both in the Netherlands) and by chemical 
GC/HRMS analysis (MAS, Münster, Germany, Nofalab, Schiedam (the Netherlands) and RIKILT, Wageningen 
(the Netherlands)). Samples of soil and samples of milk of cows and goats also have been studied by DR 
CALUX

. 
 
We also found dead birds, abandoned eggnests and thin eggscales during visual inspections of the industrial area. 
In addition to this reports of declining bird populations, inexplicable sudden deaths of domestic animals like 
goats and sheep have been given. So we decided to study the livers of hares and sheep, animals that were found 
dead in the terrain. The results will be published later. 
 
DR CALUX

 bioanalysis: The procedure for the BDS DR CALUX
 
bioassay has been described in detail 
previously
1
. Briefly, H4IIE cells stably transfected with an AhR-controlled luciferase reporter gene construct, 
were cultured in α-MEM culture medium, supplemented with 10 % (
v
/v) FCS under standard conditions (37 
o
C, 5 
% CO2, 100 % humidity). Cells were exposed in triplicate on 96-well microtiterplates containing the standard 
2,3,7,8-TCDD calibration range, a DMSO blank. Following an 24 hour incubation period cells were lysed. A 
luciferine containing solution (Glow Mix) was added and the luminescense was measured using a luminometer 
(Berthold Centro XS3). 
 
GC/HRMS analysis: all three labs, MAS (Münster, Germany), RIKILT Wageningen and Nofalab Schiedam 
(both the Netherlands) are iso 17025 accredited according EC/252 guideline. 
 
 
 
 
 
 
 
 
 
 
 
Organohalogen Compounds Vol. 76, 1407-1410 (2014) 1407 
Results and discussion 
 
The results of the DR CALUX

 bioanalyses of the eggs are shown in Figure 1. 
 
Figure 1: DR CALUX

 sum dioxins/dl-PCBs in eggs of backyard chickens near Harlingen 
 
Figure 2 gives the results of soil samples from three locations as analysed by DR CALUX

. At Midlum (HM), 
located at a distance of 750 metres (m) from the harbour, the level of dioxins/dl-PCBs in the soil was 4.45 pg 
BEQ / g product. For comparison, the level of the sum PCDD/Fs/dl-PCBs in the egg sample was 10.9 pg TEQ / 
g fat as measured by GC/HRMS. Historical land filling in the vicinity, backyard burning or emissions of 
industrial activities from the harbour could be possible sources of this pollution. 
 
 
 
Figure 2: DR CALUX

 PCDD/Fs/dl-PCBs in soil samples near Harlingen, and GC/HRMS results of Midlum (HM) egg sample 
Organohalogen Compounds Vol. 76, 1407-1410 (2014) 1408 
 
The levels of PCDD/Fs/dl-PCBs measured by DR CALUX

 in goat and cow milk at three locations near 
Harlingen correspond with typical background levels in the Netherlands (see Figure 3). 
 
 
 
Figure 3: DR CALUX

 PCDD/Fs/dl-PCBs in goat and cow milk samples near Harlingen 
 
Figure 4 gives the levels of pollution as a function of the distance to the harbour, that is to say, the DR CALUX

 
and GC/HRMS analyses for the sum of PCDD/Fs/dl-PCBs in eggs of backyard chickens near the harbour of 
Harlingen. 
 
 
 
 
Figure 4: Levels ofpollution (pg BEQ or TEQ / g fat) as a function of the distance (m) to the harbour 
Organohalogen Compounds Vol. 76, 1407-1410 (2014) 1409 
 
Figure 5 shows the levels of PCDDs, PCDFs and dl-PCBs (pg TEQ / g fat) of several egg samples near 
Harlingen as analysed by GC/HRMS. At least two different kinds of pollution source can be derived from the 
figure, a PCB source due to elevated dl-PCB and PCDF levels (samples Midlum (HM), Wijnal-17 and Wijnal-2) 
and a PCDD source due to elevated PCDD levels (samples Harl-Mid, Mid-B and Mid-L). 
 
 
 
Figure 5: Levels of PCDDs, PCDFs and dl-PCBs (pg TEQ / g fat ) of egg samples near Harlingen as analysed by GC/HRMS 
 
 
Conclusions 
 
This study shows that eggs of backyard chickens are sensitive biomonitoring parameters for dioxin/PCB 
contamination in the vicinity of potential sources (harbour activity, waste incinerator, landfill). By using cost-
efficient screening analysis tools (such as DR CALUX

) that are also affordable by private households, areas of 
public concern can be monitored in a rapid and efficient way. The correlation between DR CALUX

 and 
GC/HRMS proofed to be very satisfactory; there were no false positive or false negative results. 
The results of this study point to at least two types of source of dioxin emission near Harlingen. The fact that the 
levels of dioxins and dioxin-like PCBs are increasing with decreasing distance to the harbour urgently calls for 
closer investigation in order to find the sources of the dioxin/PCB pollutions, and to prevent adverse health 
impacts for domestic animals, wildlife and human beings. 
 
Acknowledgements 
 
This project from the NGO Toxicowatch Foundation has been funded by citizens which are concerned about 
industrial pollution in their environment. The Toxicowatch Foundation likes to thank the RIKILT Institute for 
sharing their DR CALUX

 and GC/HRMS data of egg samples. 
 
References 
1. Besselink, H., Jonas, A., Pijnappels, M., Swinkels, A. and Brouwer, B. (2004). Organohalogen Compounds, 
66: 677-681. 
Organohalogen Compounds Vol. 76, 1407-1410 (2014) 1410 
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