Changes in se um dioxin and PCB le els in esiden s a ound a municipal was e incine a o in Bilbao, Spain
MB Zube o, 1,2 E Egui aun, 1 JJ Au ekoe xea,1,2,3 A Le xundi, 1,2,4 E Abad,5 J Pa e a, 5 F Goñi-I igoyen ,2,3,4 J Iba luzea,2,3,4
(Mi en B Zube o,1,2 Elena Egui aun,1 Juan J Au ekoe xea,1,2,3 Ai ana Le xundi,1,2,4 Es eban Abad,5 Jo di Pa e a,5
Fe nando Goñi-I igoyen, 2,3,4 Jesus Iba luzea,2,3,4,6)
1 Depa men o P e en a i e Medicine and Public Heal h, Uni e si y o he Basque Coun y (UPV/EHU), Leioa, Bizkaia, Spain
2 Biodonos ia Heal h Resea ch Ins i u e, San Sebas ian, Spain
3 Public Heal h Depa men , Basque Go e nmen , San Sebas ian, Spain
4 Spanish Conso ium o Resea ch on Epidemiology and Public Heal h (CIBERESP), Spain
5 Labo a o y o Dioxins, IDAEA-CSIC, Jo di Gi ona 18, 08034 Ba celona, Spain
6 Facul y o Psychology, Uni e si y o he Basque Coun y UPV/EHU, San Sebas ian, Spain
Mi en B Zube o, Depa men o P e en a i e Medicine and Public Heal h, Uni e si y o he Basque Coun y (UPV/EHU), Leioa, Bizkaia,
Spain.
Biodonos ia Heal h Resea ch Ins i u e, San Sebas ian, Spain;
[email p o ec ed]
Elena Egui aun, Depa men o P e en i e Medicine and Public Heal h, Uni e si y o he Basque Coun y (UPV/EHU), Leioa, Bizkaia,
Spain.
[email p o ec ed]
Juan J Au ekoe xea, Depa men o P e en i e Medicine and Public Heal h, Uni e si y o he Basque Coun y (UPV/EHU), Leioa,
Bizkaia, Spain;
Public Heal h Depa men , Basque Go e nmen , San Sebas ian, Spain;
Biodonos ia Heal h Resea ch Ins i u e, San Sebas ian, Spain.
[email p o ec ed]
Ai ana Le xundi, Depa men o P e en i e Medicine and Public Heal h, Uni e si y o he Basque Coun y (UPV/EHU), Leioa, Bizkaia,
Spain;
Biodonos ia Heal h Resea ch Ins i u e, San Sebas ian, Spain;
Spanish Conso ium o Resea ch on Epidemiology and Public Heal h (CIBERESP), Spain.
[email p o ec ed]
Es eban Abad, Labo a o y o Dioxins, IDAEA-CSIC, Jo di Gi ona 18, 08034 Ba celona, Spain.
[email p o ec ed]
Jo di Pa e a, Labo a o y o Dioxins,IDAEA-CSIC, Jo di Gi ona 18, 08034 Ba celona, Spain.
[email p o ec ed]
Fe nando Goñi-I igoyen, Public Heal h Depa men , Basque Go e nmen , San Sebas ian, Spain;
Biodonos ia Heal h Resea ch Ins i u e, San Sebas ian, Spain;
Spanish Conso ium o Resea ch on Epidemiology and Public Heal h (CIBERESP), Spain.
[email p o ec ed]
Jesus Iba luzea, Public Heal h Depa men , Basque Go e nmen , San Sebas ian, Spain;
Biodonos ia Heal h Resea ch Ins i u e, San Sebas ian, Spain;
Facul y o Psychology, Uni e si y o he Basque Coun y UPV/EHU, San Sebas ian, Spain;
Spanish Conso ium o Resea ch on Epidemiology and Public Heal h (CIBERESP), Spain.
[email p o ec ed]
Co esponding au ho : Juan J Au ekoe xea, Biodonos ia Heal h Resea ch Ins i u e, San Sebas ian, Spain; Depa men o P e en a i e
Medicine and Public Heal h, Uni e si y o he Basque Coun y (UPV/EHU), Leioa, Bizkaia, Spain; Public Heal h Depa men , Basque
Go e nmen , San Sebas ian, Spain. [email p o ec ed]
Compe ing in e es s: None.
Pa ien consen : Ob ained.
E hics app o al: This s udy was conduc ed wi h he app o al o he E hics Commi ee o he Uni e si y o he Basque Coun y
Funding: This s udy was unded by he MSWP Zabalga bi S.A.
Keywo ds: O ganochlo ine Compounds; Biological Moni o ing; Incine a ion; Se um; Public Heal h Su eillance; Humans
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This is he Accep ed Manusc ip e sion o a Published Wo k ha appea ed in inal o m in En i onmen al Resea ch 156 : 738-746 (2017). To access
he inal edi ed and published wo k see h ps://doi.o g/10.1016/j.en es.2017.04.039
© 2017. This manusc ip e sion is made a ailable unde he CC-BY-NC-ND 4.0 license h ps://c ea i ecommons.o g/licenses/by-nc-nd/4.0/
ABSTRACT
The e is a g ea conce n in he Basque Coun y ega ding emissions om was e incine a ion, in pa icula
ha o o ganochlo ines (OCs), including dioxins, u ans and polychlo ina ed biphenyls (PCBs), and hei
po en ial e ec on human heal h. In 2005, a municipal solid was e plan (MSWP) s a ed o ope a e in
Bilbao, ep esen ing an oppo uni y o assess he exposu e o he a o emen ioned pollu an s among people
li ing a a ious dis ances om he plan . In 2006 and 2008, we ca ied ou wo c oss-sec ional s udies o
quan i y and assess changes in le els o hese pollu an s. The objec i e o his s udy was o desc ibe he
le els o OCs in he blood se um in 2013 o 127 adul s o his p ospec i e coho , in ou cen es o
popula ion, nea o and u he away om he MSWP, and o s udy ends o e ime since i s a ed o
ope a e. This s udy shows he le els o OCs ha e dec eased signi ican ly, by 78.9% o mo e. Fu he , le els o
OCs in a eas nea o he MSWP we e no ound o be highe han hose in a eas u he a ield.
ABBREVIATIONS
CI: con idence in e al
dl-PCB: dioxin-like PCB
GM: geome ic mean
MSWP: municipal solid was e plan
OC: o ganochlo ine
OCDD: oc achlo odibenzodioxin
PCB: polychlo ina ed biphenyl
PCDD: polychlo ina ed dibenzodioxin
PCDF: polychlo ina ed dibenzo u an
TEQ: oxic equi alency ac o
DLC: dioxin-like compounds
1. INTRODUCTION
The managemen o he solid was e we p oduce is among he g ea challenges acing socie y oday. One o
se e al me hods o educing he olume o his was e is incine a ion. Howe e , municipal solid was e
incine a ion is a po en ial sou ce o pollu ion, gi en ha du ing he p ocess he e is combus ion o esidues
con aining chlo ine, as well as o hea y me als (Van Caneghem e al., 2010). Du ing combus ion p ocesses,
he e is a elease o a ious pollu an s such as o ganochlo ines, including he polychlo ina ed-p-
dibenzodioxins, he polychlo ina ed dibenzo u ans, commonly known as dioxins and u ans o simply dioxin
like compounds (DLCs), and polychlo ina ed biphenyls (PCBs). Indeed, combus ion is he main sou ce o DLCs
eleased in o he en i onmen (Domingo e al., 2000). Speci ically, among he mal sou ces, solid was e
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incine a ion has been iden i ied as he p incipal sou ce o DLCs du ing he 1980s and ea ly 1990s (Jones e
al., 1993; Eduljee e al., 1996; Quaß e al., 2000; Kim e al., 2001; Quaß e al., 2004; Bu ns e al., 2010). Since
hen, municipal solid was e plan s (MSWPs) ha e become a mo e e icien me hod o ea ing solid was e
and mo e e ec i e a con olling DLC emissions (Ranzi e al., 2011).
Ne e heless, he e is s ill conce n ega ding he en i onmen al and heal h consequences o was e
incine a ion, and hence, i is impo an o assess i s impac a he en i onmen al le el and on people li ing
a ound MSWPs. A sys ema ic e iew concluded ha he le el o e idence o MSWPs ha ing an e ec on
heal h was limi ed o all ypes o cance s, so issue sa coma, and ep oduc i e and bi h de ec s (o o acial
and geni ou ina y) and inadequa e o all he o he heal h p oblems conside ed (Ma iello e al., 2013).
The opening o a MSWP in Bilbao in 2005 ep esen ed a g ea oppo uni y o assess exposu e o ce ain
pollu an s among people li ing a a ious dis ances om he plan . In 2006 and 2008, wo c oss-sec ional
s udies we e ca ied ou o quan i y and assess changes in he le els o OCs in a coho o he gene al u ban
popula ion, hose wi h no wo k- ela ed exposu e and li ing in a eas nea o and u he away om he
MSWP (Zube o e al., 2009; Zube o e al., 2011). Gi en he high oxici y o dioxins and PCBs, a s udy was
ca ied ou o e i y i he e was a heal h isk o hose close o he MSWP. The objec i e o s udying his new
c oss-sec ion o he coho was o assess he le els o DLCs in people in ou cen es o popula ion, nea o
and u he away om he MSWP and s udy ends o e ime since i s a ed o ope a e.
2. MATERIAL AND METHODS
We ca ied ou p ospec i e coho s udy wi h h ee c oss-sec ional analyses among people li ing in a eas
nea o and u he away om he MSWP. The sampling was pe o med in 2006 (Zube o e al., 2009), 2008
(Zube o e al., 2011) and in he hi d qua e o 2013. The i s s age o he s udy was conduc ed when he
MSWP s a ed o ope a e, and he da a collec ed hen is conside ed he baseline o subsequen s ages o
he s udy.
2.1.STUDY POPULATION
Rega ding dis ance o he MSWP, we selec ed ou a eas: wo a eas nea by, Alonso egi and Rekalde, and
wo a eas u he away, San u xu and Balmaseda, (Zube o e al., 2011). The a eas conside ed close o he
MSWP we e he own o Alonso egi and he bo ough o Rekalde in Bilbao, bo h less han 2 km away om
he plan and in u ban en i onmen s wi h high a ic densi y. The a eas u he away, used as con ol a eas,
we e he bo ough o San u xu in Bilbao, also an u ban en i onmen wi h high a ic densi y (5 km away
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om he plan ) and a small own wi h low a ic densi y, Balmaseda (20 km away om he plan ); bo h o
hese a eas ha e li le indus ial ac i i y and a e ou o he pa h o p e ailing winds.
In ela ion o he o igin o he samples, in 2006, 36% o he pa icipan s came om he census in he a eas
close o he plan , and he es we e olun ee s om he heal h cen e. In emo e a eas, 25% we e ec ui ed
om he census, and he es we e olun ee s. In 2008, in nea by a eas, 71% we e p e ious pa icipan s,
whe eas in he emo e a eas his pe cen age was 68%. In 2013, we ca ied ou he hi d s age o he s udy,
ec ui ing people who had pa icipa ed in he wo p e ious s udies, wi h he aim o ec ui ing 30 people
om each s udy a ea, s a i ied by sex and age g oup a baseline (20-69 yea s old). The exclusion c i e ia a
all s ages we e: ha ing li ed in he a ea o less han he p e ious 5 yea s, ha ing wo ked in a company
which ca ied ou p oduc ion p ocesses in ol ing po en ial exposu e o DLCs in he p e ious 15 yea s, and
being 70 yea s old o abo e.
Ha ing elephoned 223 candida e pa icipan s, we inally ob ained blood samples om 127 indi iduals. The
main easons why pa icipan s in ea lie s ages o he s udy did no pa icipa e in his s age we e wo k
commi men s, ill heal h/dea h, age, o declining u he pa icipa ion, as well as no being possible o
con ac hem, o ha hey had mo ed away.
All pa icipan s signed an in o med consen o m o ake pa in he s udy, which was app o ed by he E hics
Commi ee o Resea ch and Teaching o he Uni e si y o he Basque Coun y.
2.2.STUDY VARIABLES
We in e iewed all pa icipan s and collec ed da a on an h opome ic and socioeconomic cha ac e is ics;
ep oduc i e and b eas eeding his o y, in ake o local ood p oduce, and smoking and d inking habi s.
Weigh gain o loss o mo e han 5 kg in he p e ious 5 yea s was no ed. Rega ding social class, pa icipan s
we e classi ied in o one o wo ca ego ies, manual o non-manual wo ke s, acco ding o hei mos ecen
occupa ion (no as possible sou ce o DLC bu as possible con ounding ac o ), o ha o he head o he
amily i hey did no wo k, in line wi h he Spanish Adap a ion o he B i ish OPCS Classi ica ion o
Occupa ions (social classes I+II+III and IV+V). Pa icipan s we e also classi ied in o wo g oups by le el o
educa ion, depending on whe he o no hey had uni e si y quali ica ions. All pa icipan s we e in e iewed
by he same esea che o a oid in e iewe bias. We collec ed da a on he equency o in ake o local ood
p oduce ( ui and ege ables, cow's milk and cheese, poul y and o he mea ), wi h ou esponse op ions:
ne e o less han once e e y 2 mon hs, occasional (a ound once a mon h), a ound once a week, o daily
(mo e han wice a week). These da a we e ecoded o a dicho omous a iable: ne e o occasionally e sus
weekly o daily in ake.
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2.3.BLOOD TEST ANALYSIS
A 70-ml blood sample was collec ed om all s udy pa icipan s. They we e no equi ed o as , bu we e
ecommended o a oid ea ing a y oods he nigh be o e. Blood samples we e collec ed in blood collec ion
ubes (Vacu aine ®) wi hou an icoagulan and ans e ed immedia ely o a glass ube o cen i uga ion. To
ob ain he se um, he blood was le o clo o app oxima ely 60-75 minu es a oom empe a u e. The
sample was hen cen i uged a 1500 G o 15 minu es. The se um was sepa a ed om he clo wi h a
Pas eu pipe e. Unlike in p e ious s ages o he s udy, in which he analysis was pe o med using pools o
samples g ouped by age and sex, he analysis in his s udy was based on indi idual samples. Samples we e
ozen a -20°C wi hin 90 minu es a e collec ion. Blood ex ac ion, and se um p epa a ion, s o age and
anspo we e ca ied ou in acco dance wi h he condi ions es ablished by he e e ence labo a o ies
(Pa e son e al., 1991).
Nex , wo k low analysis is pe o med as ollows: abou 40 mL o se um was spiked wi h a 13C12 labelled
dioxin and PCB s anda d mix u e. A e wa ds, analy es we e ex ac ed om he ma ix by solid-phase
ex ac ion (SPE) app oach using C18 ca idges (SPE Ca idges – C18; ISOLUTE, Cha lo e, NC, USA). Elu ion
was pe o med wi h n-hexane. O ganic componen s, a and o he in e e ing subs ances we e emo ed by
ea ing he hexane ex ac s wi h silica gel (Supelco, Belle on e, PA, USA) modi ied wi h sulphu ic acid (44%).
Fu he , he ex ac s we e cleaned up by solid-liquid adso p ion ch oma og aphy, using mul ilaye silica,
basic alumina and ac i a ed ca bon adso ben s. Pu i ied ex ac s we e o a y concen a ed, ans e ed in o
ials and concen a ed o d yness by a gen le s eam o ni ogen p io o he mass spec ome y analysis. To
e alua e he eco e y a es, inal ex ac s we e econs uc ed in a known amoun o a mix u e o labelled
13C12-PCDD/Fs (EPA-1613ISS, Welling on Labo a io es Inc., Guelph, Canada), 13C12-dl-PCBs (WP-ISS,
Welling on Labo a o ies Inc, Guelph, Canada) and 13C12-ma ke PCBs (MBP-118, Welling on Labo a o ies Inc,
Guelph, Canada).
Ins umen al analysis was based on he use o high esolu ion gas ch oma og aphy coupled o a high
esolu ion mass spec ome y (HRGC-HRMS). Analyses we e pe o med on an Agilen gas ch oma og aph
(Agilen Technologies, Palo Al o, CA, USA) coupled o an Au oSpec P emie high esolu ion mass
spec ome e (Wa e s, Manches e , UK) a 10,000 esol ing powe (10% alley de ini ion). Gas
ch oma og aphic sepa a ion was pe o med on a DB-5ms used silica column (60 m x 0.25 mm i.d. x 0.25-µm
ilm hickness; J&W Scien i ic, CA, USA) o PCDD/Fs and dl-PCBs, whe eas o ma ke PCBs, he analysis was
ca ied ou on a DB-XLB used silica column (60 m x 0.25 mm i.d. x 0.25 µm- ilm hickness; J&W Scien i ic, CA,
USA). Quan i ica ion was ca ied ou by he iso opic dilu ion me hod. Rela i e esponse ac o s we e
measu ed o each indi idual compound by he analysis o six di e en calib a ion solu ions o PCDDs/Fs, dl-
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PCBs and ma ke PCBs. Finally, he esul s we e exp essed as TEQ ( oxic equi alen ) alues using WHO-TEQ
(Wo ld Heal h O ganiza ion Toxic Equi alen ). WHO-TEQ alues we e calcula ed in "uppe bound", assuming
he limi o de ec ion (LOD) o hose non-de ec ed o below he LOD congene s. Mo e de ailed in o ma ion
on ins umen al analysis is epo ed elsewhe e (Abad e al., 2000).
Enzyma ic lipid de e mina ions we e pe o med using 100 µL se um sub-sample. O e all, ou ypes o lipids
we e conside ed: iglyce ides, o al choles e ol, non-es e i ied ( ee) choles e ol and phospholipids. A well-
documen ed summa ion me hod was used o calcula e he o al lipid concen a ions (Pa e son e al., 1991).
The o al lipid con en was exp essed in g/L. Fo he con e sion be ween olume ic and g a ime ic da a, a
alue o 1.026 g/mL was used o se um speci ic g a i y. The equi emen s o ensu ing high quali y da a
include he applica ion o quali y assessmen and quali y con ol measu es. A se ies o con ol blanks we e
analysed o assess labo a o y c oss-con amina ion. De ailed in o ma ion on he analy ical me hod is
included as SI.
2.4.STATISTICAL METHODS
The sample was desc ibed using he absolu e and ela i e equencies o he a iables collec ed du ing he
in e iew. The le els o congene s we e desc ibed by mean, median, and ange (minimum and maximum
alues), assessing hei con ibu ion o he WHO-TEQs. Pea son’s coe icien o co ela ion be ween he
le els o congene s was calcula ed. Fu he , he geome ic means (GMs) and 95% con idence in e als (CIs)
o he lipid-co ec ed da a we e calcula ed o each g oup o pollu an s and compa ed as a unc ion o
socioeconomic and ep oduc i e a iables.
Fo he empo al analysis o he le els o PCDD/Fs and dl-PCBs, gi en ha he p e ious samples we e
analysed in se um pools, he co esponding alue o a new pool was calcula ed using he a i hme ic mean o
each indi idual alue in 2013, because he measu ed concen a ion in a pooled sample is compa able o he
a i hme ic mean o indi idual specimens making up he pool (He e nan e al., 2014). The pooled samples o
2006, 2008 and 2013 we e adjus ed o he no mal dis ibu ion. Likewise, he dis ibu ions o he di e en
g oups o OCs, a e loga i hmic ans o ma ion, analyzed on indi idual samples we e adjus ed o he no mal
(Kolmogo o -Smi no Z es p> 0.10). Changes in he a i hme ic mean le els o dioxins, u ans, and dl-PCBs
in he pools om 2006 o 2008 and om 2006 o 2013 we e analysed using a pai ed - es . Simila ly, changes
in he geome ic means o ma ke PCBs (indi idual samples) o e hese pe iods we e analysed using a pai ed
- es .
A mul iple linea eg ession analysis was pe o med o iden i y he socioeconomic a iables associa ed wi h
each g oup o con aminan s. All a iables we e included in any o he models wi h a p <0.10 acco ding o he
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likelihood a io es . All he models included he geog aphical loca ion, age and sex a iables, since hey
we e ela ed o he selec ion o he sample in p e ious s ages. Resul s a e exp essed as a pe cen age change
[(exp(b)-1) * 100]. The ela ion be ween OCs and in ake o local ood p oduce was plo ed, aking in o
accoun he dis ance o he MSWP. The c i e ion o s a is ical signi icance was p< 0.05. The s a is ical
analysis was ca ied ou using IBM SPSS S a is ics o Windows, Ve sion 20.0 (IBM SPSS, A monk, NY, USA).
3. RESULTS
Table 1 shows he cha ac e is ics o he sample. Sligh ly mo e han hal o he 127 pa icipan s we e women.
Ca ego izing age in o e iles, app oxima ely a hi d o pa icipan s we e unde 47 yea s o age, ano he hi d
we e be ween 47 and 60 and he las hi d o e 60 yea s o age. The highes pa icipa ion a e was obse ed
in Balmaseda. A o al o 29.1% o pa icipan s had no mal weigh , and a simila pe cen age was o e weigh ,
while 35.4% was obese. Nea ly hal o he sample epo ed weigh change in he p e ious 5 yea s, 35.4%
ha ing pu on weigh and 14.2% ha ing los weigh (≥ 5 kg). Rega ding occupa ion and educa ion, 42.5% o
pa icipan s we e manual wo ke s and 28.3% had uni e si y quali ica ions. O e all, 20% we e smoke s a he
ime o he in e iew, 34.6% we e ex-smoke s and 43.3% we e ne e -smoke s. Among he women, mo e
han hal we e in menopause, while 76.1% had had child en and 77.8% had b eas ed. Compa ed wi h
p e ious yea s, in 2013, in addi ion o he co esponding inc ease in age, he e was an inc ease in BMI and,
also an inc ease in he numbe o women who had child en. Social class showed non-manual wo ke s o a
g ea e ex en in he 2013 sample han in he p e ious wo. The e we e no signi ican di e ences in he
cha ac e is ics o he es o he a iables.
Table 2 shows he geome ic mean, median, minimum and maximum le els o each o he congene s
s udied, as well as he WHO-TEQ alues and he weigh o each in he o al le el o subs ances wi h dioxin
ac i i y (DLC). Al hough a highe concen a ion o mono-o ho-PCBs was obse ed, especially PCB 118, hey
because o hei lowe oxici y, con ibu ed a small pa o he o al WHO TEQ. Ma ke PCBs showed a clea
dec ease in le els, lowe han DLCs. Th ee compounds, PCB 126, 12378-PeCDD (1,2,3,7,8-
pen achlo odibenzo-p-dioxin) and 23478-PeCDF (2,3,4,7,8-pen achlo odibenzo u an), accoun ed o 62.81%
o he o al WHO-TEQs. In con as , o he compounds wi h a high concen a ion in blood se um such as
mono-o ho PCBs and o a lesse ex en oc achlo odibenzodioxin (OCDD) and PCB77 con ibu ed e y li le
o he WHO-TEQs, due o hei low oxic equi alency ac o . Supplemen a y Table 1 shows he co ela ions
obse ed be ween he congene s; he coe icien s being he highes among he PCBs and low be ween hese
and bo h dioxins and u ans, while he e was an in e media e co ela ion be ween dioxins and u ans.
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
378
379
380
381
382
383
384
385
386
387
388
389
390
391
392
393
394
395
396
397
398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
413
Table 3 shows a d ama ic dec ease in he mean alues o OCs by 2013 compa ed o 2008 and 2006, in he
o al and in he ou s udied a eas. The e was no a signi ican dec ease in mean alues be ween 2006 and
2008. Indeed, in his pe iod, we ound a signi ican inc ease in he concen a ion o dioxin-like PCBs.
Table 4 lis s he GMs and 95% CIs o each pollu an wi h espec o he socioeconomic, ep oduc i e and
b eas eeding a iables. Men had signi ican ly highe le els o dl-PCBs han women. Among he ou s udy
a eas, we ound he highes mean OC le els in San u xu, he di e ence being signi ican in all cases excep
o PCDD/Fs. We obse ed ha indi iduals wi h uni e si y quali ica ions had signi ican ly lowe mean le els
o dioxins, bu highe le els o dl-PCBs. We did no obse e signi ican di e ences be ween OCs and social
class, body mass index (BMI) o weigh change (da a no shown). Le els o dl-PCBs we e highe in women
who had had child en han hose who had no , he di e ence being ma ginally signi ican ; while hose who
had b eas ed had lowe le els o ma ke PCBs, he di e ence again being ma ginally signi ican . Pos -
menopausal women had signi ican ly highe le els o dl-PCBs han o he women.
Mul iple linea eg ession analysis (Table 5) showed ha compa ed o he coe icien s o Balmaseda, which
is he leas u ban and indus ialised o he a eas s udied and he u hes om he incine a o incine a o ,
he highes coe icien s o all OCs we e ound o San u xu, he di e ences being s a is ically signi ican o
dl-PCBs, DLC and ma ke PCBs, bu no o PCDD/Fs. In he o he wo zones, hose closes o he MSWP, he
coe icien s did no show di e ences om hose o Balmaseda, hough Alonso egi showed he lowes
coe icien s o all he OCs. Compa ing he sexes, men did no show di e ences in ma ke PCB le els wi h
women. Le els o bo h ypes o PCBs we e posi i ely associa ed wi h age; PCDD/Fs le els, howe e , we e no
signi ican ly associa ed wi h age.
Indi iduals who had only comple ed non-uni e si y educa ion had signi ican ly highe le els o PCDD/Fs han
pa icipan s wi h uni e si y quali ica ions. The le els o PCDD/Fs and DLC we e in e sely ela ed o weigh
gain in he p e ious 5 yea s, bu he di e ences we e no signi ican . Women who had b eas ed had
signi ican ly lowe le els o PCBs han he es o all he pa icipan s.
Figu e 1 shows he ela ionship o OC le els wi h in ake o local ood p oduce and dis ance om he MSWP
s udied (Rekalde and Alonso egi being nea by, while Balmaseda and San u xu a e u he away). We did no
obse e any signi ican inc eases in he mean le els o OCs wi h in ake o local ood p oduce in a eas nea
he MSWP.
4. DISCUSSION
414
415
416
417
418
419
420
421
422
423
424
425
426
427
428
429
430
431
432
433
434
435
436
437
438
439
440
441
442
443
444
445
446
447
448
449
450
451
452
453
454
455
456
457
458
459
460
461
462
463
464
465
466
467
468
469
470
471
472
In ou s udy, indi iduals li ing nea o a solid was e MSWP did no ha e highe blood le els o OCs han
hose li ing u he a ield. On he con a y, he highes le els o hese pollu an s we e ound in one he mo e
dis an a eas, e en a e adjus ing o con ounding ac o s. Fu he , he in ake o local ood p oduce nea he
MSWP was no associa ed wi h an inc ease in he blood concen a ion o he pollu an s analysed. These da a
a e in ag eemen wi h hose epo ed by Ranzi e al. (2011), who ound ha he a io o emissions in 2008 o
hose in 1994-1996 o wo I alian incine a o s was abou 0.0001 (10,000 imes less) o dioxins and u ans.
O he s udies analysing ends o e ime in a eas close o MSWPs made simila obse a ions, wi h no
inc ease in OC le els in a eas close o MSWPs (Deml e al., 1996; E ans e al., 2000; González e al., 2000;
Chen e al., 2004; Huang e al., 2007; Reis e al., 2007; De Felip e al., 2008; Pa e a e al., 2013).
In addi ion, we obse ed signi ican dec eases in he le els o OCs, bo h in he a eas nea o and u he
away om he plan be ween 2006 o 2008 and 2013. Wi siepe e al. (2000) also obse ed a dec ease om
43.77 o 20.7 pg TEQ/g lipid be ween 1989 and 1998, and Humble e al. (2011) a dec ease om 36 o 25 pg
TEQ/g lipid in women be ween 2000 and 2009. Simila ly, Po a e al. (2012) epo ed OC le els o ha e
dec eased by 34 o 56% be ween 2002 and 2006, and Llobe e al. (2003) also ound a dec ease in OC le els
o e ime. A gene al endency o PCB le els o dec ease was also obse ed by Linde holm e al. (2010)
be ween 1990 and 2007 and Humble e al. (2011), who ound a dec ease om 291 o 211 ng/g lipid in
women. A sys ema ic e iew o s udies published up o 2010 on he blood le els o dioxins, u ans and dl-
PCBs epo ed a mean annual dec ease o 7% o dioxins and 6% o bo h u ans and dl-PCBs (Consonni e
al., 2012). A s udy based on he incine a o in Ma a ó, Spain (Pa e a e al., 2013) ound a dec ease in he
le els o PCDD/Fs and ma ke PCBs, as in ou s udy. A ecen s udy showed also an impo an dec ease o
se um PCBs in a popula ion li ing nea a con amina ed a ea a e public heal h in e en ion (Magoni e al.,
2016).
Unlike a ious o he s udies (Deml e al., 1996; E ans e al., 2000; González e al., 2000; Reis e al., 2007), we
did no ind signi ican di e ences be ween he le els o PCDD/Fs by age. On he o he hand, dl-PCB and
ma ke PCB le els we e posi i ely associa ed wi h age. The a o emen ioned sys ema ic e iew o blood OC
le els, showed a 2% inc ease in le els o he h ee g oups o OCs (PCDDs, PCDFs and dl-PCBs) wi h each yea
o age (Consonni e al., 2012). A p e ious s udy o samples collec ed om ou s udy popula ion be ween
2006 and 2008 (Zube o e al., 2015) showed an inc ease in he le els o OC pes icides and ma ke PCBs wi h
age, consis en wi h a coho e ec , and a dec ease be ween samples aken in he wo pe iods (a dec easing
pe iod e ec ). This sugges s ha he highe le els ound in olde indi iduals a e due o he ac ha OC
exposu e has dec eased o e he yea s, and hence has less impac on younge coho s, a he han a p ocess
o bioaccumula ion o e he yea s.
473
474
475
476
477
478
479
480
481
482
483
484
485
486
487
488
489
490
491
492
493
494
495
496
497
498
499
500
501
502
503
504
505
506
507
508
509
510
511
512
513
514
515
516
517
518
519
520
521
522
523
524
525
526
527
528
529
530
531
Supplemen a y Table 1: Pea son’s co ela ion coe icien be ween di e en congene s (dioxins, u ans, dl-PCBs and ma ke PCBs).
2378TCDD
12378PeCDD
123478HxCDD
123678HxCDD
123789HxCDD
1234678HpCDD
OCDD
2378TCDF
12378PeCDF
23478PeCDF
123478HxCDF
123678HxCDF
234678HxCDF
123789HxCDF
1234678HpCDF
1234789HpCDF
OCDF
PCB77
PCB81
PCB126
PCB169
PCB105
PCB114
PCB118
PCB123
PCB156
PCB157
PCB167
PCB189
PCB28
PCB52
PCB101
PCB138
PCB153
PCB180
2,3,7,8-TCDD
1
1,2,3,7,8-PeCDD
.33
1
1,2,3,4,7,8-HxCDD
.49
.50
1
1,2,3,6,7,8-HxCDD
.26
.38
.31
1
1,2,3,7,8,9-HxCDD
.39
.55
.61
.51
1
1,2,3,4,6,7,8-HpCDD
.25
.45
.43
.71
.53
1
OCDD
.18
.33
.29
.65
.44
.66
1
2,3,7,8-TCDF
.41
.32
.27
.13
.16
.18
.12
1
1,2,3,7,8-PeCDF
.34
.40
.35
.21
.29
.25
.18
.34
1
2,3,4,7,8-PeCDF
.23
.50
.28
.69
.42
.59
.58
.18
.39
1
1,2,3,4,7,8-HxCDF
.33
.51
.50
.44
.52
.54
.39
.33
.41
.40
1
1,2,3,6,7,8-HxCDF
.32
.42
.42
.34
.41
.44
.31
.25
.46
.37
.67
1
2,3,4,6,7,8-HxCDF
.34
.40
.44
.33
.45
.33
.28
.32
.53
.38
.56
.49
1
1,2,3,7,8,9-HxCDF
.32
.40
.49
.19
.44
.20
.13
.16
.46
.24
.50
.42
.44
1
1,2,3,4,6,7,8-HpCDF
.25
.47
.42
.55
.44
.62
.53
.31
.35
.60
.53
.45
.44
.39
1
1,2,3,4,7,8,9-HpCDF
.40
.41
.46
.22
.44
.23
.15
.36
.50
.21
.44
.45
.61
.55
.36
1
OCDF
.28
.45
.37
.19
.46
.20
.12
.28
.44
.19
.38
.29
.42
.49
.29
.61
1
PCB-77
.23
.13
.19
.26
.12
.27
.30
.13
.16
.24
.14
.25
.17
.07
.23
.13
-.08
1
PCB-81
.18
.10
.18
.15
.10
.07
.076
.04
.01
.13
.01
.05
.03
.11
.12
.11
-.03
.68
1
PCB-126
.14
.09
.21
.23
.06
.17
.17
-.01
-.11
.20
.01
.07
-.02
-.03
.13
-.05
-.10
.49
.67
1
PCB-169
.23
.16
.24
.45
.21
.27
.30
-.02
-.05
.39
.11
.11
.09
.13
.28
.07
-.08
.51
.79
.73
1
PCB-105
.19
.24
.12
.56
.23
.55
.56
.15
.10
.47
.27
.32
.08
-.04
.33
-.01
-.16
.66
.36
.44
.42
1
PCB-114
.25
.25
.15
.69
.29
.61
.64
.10
.10
.60
.28
.30
.12
.04
.42
.02
-.09
.58
.38
.49
.55
.91
1
PCB-118
.19
.23
.11
.60
.25
.57
.59
.13
.09
.50
.27
.32
.06
-.04
.36
-.02
-.16
.61
.33
.44
.44
.99
.93
1
PCB-123
.22
.25
.30
.46
.34
.47
.46
.03
.10
.40
.30
.34
.05
.14
.29
.05
-.03
.65
.64
.62
.64
.72
.74
.73
1
PCB-156
.20
.24
.11
.71
.30
.58
.67
.11
.07
.62
.27
.26
.07
.03
.45
-.01
-.04
.40
.22
.39
.48
.82
.92
.88
.60
1
PCB-157
.21
.25
.13
.71
.32
.60
.67
.10
.06
.62
.29
.26
.07
.06
.46
.00
-.04
.40
.25
.41
.51
.83
.94
.88
.63
.99
1
PCB-167
.18
.25
.11
.68
.28
.59
.65
.10
.03
.59
.27
.28
.05
-.02
.41
-.04
-.11
.44
.23
.41
.49
.86
.91
.91
.64
.96
.95
1
PCB-189
.25
.24
.17
.65
.29
.54
.64
.12
.06
.58
.27
.24
.05
.09
.44
.04
-.03
.41
.27
.44
.54
.75
.85
.79
.60
.92
.91
.87
1
PCB-28
.30
.20
.17
.47
.22
.48
.52
.21
.20
.48
.23
.31
.23
.03
.44
.09
-.10
.80
.39
.38
.42
.77
.70
.74
.59
.61
.60
.62
.59
1
PCB-52
.31
.16
.22
.39
.17
.38
.42
.17
.19
.39
.16
.23
.19
.03
.35
.07
-.12
.86
.51
.44
.46
.72
.65
.68
.61
.52
.52
.54
.53
.94
1
PCB-101
.25
.10
.17
.30
.13
.32
.37
.14
.16
.30
.12
.20
.14
-.03
.24
.04
-.15
.87
.52
.44
.41
.74
.63
.69
.62
.48
.48
.51
.48
.90
.97
1
PCB-138
.24
.23
.17
.64
.30
.54
.68
.10
.08
.57
.25
.27
.09
-.02
.38
.01
-.10
.50
.26
.40
.49
.86
.88
.89
.63
.90
.90
.90
.84
.73
.65
.64
1
PCB-153
.24
.22
.17
.65
.30
.54
.68
.09
.09
.59
.25
.27
.10
-.01
.40
.00
-.10
.48
.24
.40
.49
.84
.88
.88
.62
.92
.91
.91
.86
.73
.65
.63
.99
1
PCB-180
.24
.19
.17
.64
.30
.51
.68
.09
.09
.59
.21
.23
.08
.01
.40
-.01
-.07
.45
.25
.42
.51
.78
.85
.83
.60
.92
.91
.89
.88
.70
.63
.60
.96
.98
1
Bold: p< 0.05
1
Table 4: Mean and 95% con idence in e als o le els o PCDD/Fs, dl-PCBs, DLC (pg WHO-TEQ/g a ) and o al
ma ke PCBs (ng/g a ) exp essed as pg WHO-TEQ/g a and ma ke PCBs exp essed as ng/g a , as a unc ion o
socioeconomic cha ac e is ics. 2013
PCDD/Fs
dl-PCBs
DLC
Ma ke PCBs
N
GM (CI)
GM (CI)
GM (CI)
GM (CI)
TOTAL
127
4.66 (4.22-5.14)
2.53 (2.15-2.98)
7.93 (7.21-8.73)
32.36 (25.75-40.66)
Sex
Male
56
4.75 (4.13-5.46)
3.17 (2.54-3.96)
8.66 (7.58-9.89)
35.00 (24.59-49.82)
Female
71
4.58 (3.98-5.27)
2.12 (1.69-2.66)
7.40 (6.47-8.47)
30.41 (22.52-41.07)
P
.731
.016
.112
.551
Age, yea s
< 47
42
4.69 (4.05-5.43)
2.29 (1.71-3.06)
7.69 (6.62-8.92)
26.43 (17.35-40.25)
47-60
43
4.65 (3.86-5.60)
2.54 (1.87-3.44)
7.82 (6.44-9.50)
35.23 (23.23-53.42)
> 60
42
4.63 (3.85-5.56)
2.79 (2.17-3.59)
8.30 (7.14-9.66)
36.31 (25.68-51.35)
p alue
.916
.341
.524
.271
Municipali y
Alonso egi
30
4.48 (3.74-5.36)
1.44 (1.15-1.80)
6.23 (5.38-7.22)
19.27 (12.96-28.64)
Bilbao - Rekalde
30
4.55 (3.83-5.40)
2.82 (2.12-3.76)
8.21 (7.07-9.54)
26.48 (17.74-39.53)
Bilbao - San u xu
30
4.96 (3.76-6.54)
6.18 (4.86-7.85)
11.55 (9.13-14.60)
93.75 (55.66-157.92)
Balmaseda
37
4.65 (3.94-5.48)
1.77 (1.32-2.38)
6.91 (5.88-8.13)
24.46 (17.59-34.02)
p alue
.910
.000
.000
.000
Body mass index,
kg/m2
< 25
37
4.94 (4.06-6.02)
2.81 (2.05-3.86)
8.52 (7.01-10.35)
33.90 (21.69-52.98)
25-29
45
4.76 (4.06-5.59)
2.72 (2.07-3.56)
8.13 (6.87-9.62)
34.51 (22.88-52.03)
> 30
45
4.33 (3.67-5.11)
2.16 (1.66-2.83)
7.30 (6.35-8.39)
29.20 (20.73-41.13)
p alue
.291
.199
.203
.595
Weigh loss ≥ 5 kg
Yes
63
4.68 (4.19-5.24)
2.43 (2.03-2.92)
7.84 (7.02-8.75)
33.04 (25.50-42.80)
(p e ious 5 yea s)
No
64
4.48 (3.71-5.41)
3.21 (2.33-4.43)
8.52 (7.47-9.72)
28.52 (19.52-41.67)
p alue
.764
.248
.555
.662
Weigh gain ≥ 5 kg
Yes
45
4.89 (4.34-5.50)
2.61 (2.15-3.16)
8.22 (7.35-9.21)
31.45 (23.87-41.43)
(p e ious 5 yea s)
No
82
4.26 (3.56-5.09)
2.40 (1.78-3.24)
7.42 (6.23-8.85)
34.07 (22.66-51.24)
p alue
.195
.643
.318
.744
Le el o educa ion
P e-uni e si y
91
4.96 (4.43-5.56)
2.27 (1.87-2.76)
7.96 (7.14-8.89)
29.74 (22.71-38.94)
Uni e si y
36
3.96 (3.27-4.80)
3.32 (2.48-4.43)
7.85 (6.44-9.56)
40.03 (26.09-61.44)
p alue
.044
.041
.891
.252
Social class
Manual
54
4.87 (4.22-5.62)
2.54 (1.97-3.27)
8.26 (7.24-9.41)
29.68 (21.15-41.65)
Non manual
73
4.50 (3.92-5.16)
2.52 (2.03-3.13)
7.70 (6.72-8.82)
34.49 (25.31-47.00)
p alue
.439
.967
.483
.526
In ake o local
p oduces
Ne e , occasionally
37
4.64 (2.48-6.80)
2.57 (0.25-4.89)
8.02 (5.86-10.19)
33.09 (30.61-35.58)
Weekly, daily
90
4.66 (2.58-6.74)
2.52 (0.36-4.67)
7.89 (5.82-9.97)
31.81 (29.57-34.05)
p alue
.965
.907
.880
.878
Had had child en
No
17
4.72 (3.68-6.06)
2.09 (1.31-3.33)
7.34 (5.59-9.64)
31.83 (17.88-56.68)
Yes
54
4.54 (3.84-5.37)
2.13 (1.64-2.76)
7.42 (6.35-8.67)
29.97 (21.05-42.69)
p alue
.915
.055
.283
.827
Had b eas ed
No
12
4.22 (2.96-6.04)
2.79 (1.71-4.54)
7.43 (5.20-10.62)
55.30 (28.54-107.15)
Yes
42
4.64 (3.82-5.62)
1.97 (1.45-2.67)
7.42 (6.23-8.83)
25.16 (16.84-37.60)
p alue
.657
.283
.994
.069
Menopause
No
29
4.31 (3.58-5.20)
1.71 (1.19-2.47)
6.64 (5.50-8.01)
24.87 (15.32-40.38)
Yes
42
4.78 (3.91-5.84)
2.46 (1.85-3.26)
7.98 (6.63-9.60)
34.94 (23.86-51.15)
p alue
.719
.014
.108
.474
Table 5: Mul iple linea eg ession including a iables associa ed wi h he blood le els o PCDD/Fs, dl-PCBs, DLC (pg WHO-TEQ/g a ) and ma ke PCBs (ng/g a ). Pe cen age
change and 95% con idence in e als.
PCDD/Fs
dl-PCBs
DLC
Ma ke PCB
Geog aphical a eas
Alonso egi
-7.6* (-30.6 o 22.9)
-22.0 (-46.3 o 13.3)
-12.8 (-32.0 o 11.9)
-16.5 (-53.5 o 49.7)
Bilbao - Rekalde
-7.4 (-30.0 o 22.5)
48.1 (2.8 o 113.3)
12.0 (-12.2 o 42.9)
16.6 (-34.2 o 106.5)
Bilbao - San u xu
25.9 (-6.6 o 69.8)
263.0 (145.7 o 436.2)
90.3 (46.6 o 146.9)
357.3 (148.3 o 742.2)
Balmaseda
Re e ence
Re e ence
Re e ence
Re e ence
Sex
Female
Re e ence
Re e ence
Re e ence
Re e ence
Male
1.3 (-21.8 o 31.0)
-7.7 (-34.1 o 29.2)
-3.9 (-23.3 o 20.3)
49.3 (-11.9 o 153.0)
Age
(yea s)
-0.1 (-1.1 o 0.8)
1.7 (0.5 o 2.9)
0.5 (-0.3 o 1.3)
2.6 (0.6 o 4.5)
Le el o educa ion
Non-uni e si y
Re e ence
Re e ence
Re e ence
Re e ence
Uni e si y
-32.7 (-47.8 o -13.2)
0.9 (-27.6 o 40.7)
-22.1 (-37.6 o -2.8)
-16.7 (-50.5 o 40.0)
Body mass index
(kg/m2)
-1.6 (-3.4 o 0.2)
-2.3 (-4.6 o -0.01)
-1.4 (-3.0 o 0.2)
-4.6 (-8.2 o -1.0)
Weigh gain in p e ious 5 yea s
No
Re e ence
Re e ence
Re e ence
Re e ence
Yes
-17.1 (-33.3 o 3.0)
-19.4 (-39.3 o 7.1)
-17.3 (-31.5 o 0.0)
-11.0 (-42.9 o 38.8)
Women who had b eas ed
No
Re e ence
Re e ence
Re e ence
Re e ence
(compa ed o he all es o subjec s)
Yes
-2.6 (-26.0 o 28.2)
-27.8 (-49.6 o 3.3)
-7.4 (-27.1 o 17.6)
-50.5 (-71.8 o -13.3)
* % change= (exp(b)-1) * 100; pe cen age change in OC le els associa ed wi h a change in he e e ence o he cu en ca ego y o an inc ease o one uni in he con inuous
a iables.
Hihgligh s:
- This s udy shows a d ama ic dec ease in he mean alues o OCs om 2006 and 2008 o 2013
- We ound di e ences in mean OC le els in ela ion o de a ea o sample
- Men had signi ican ly highe le els o dl-PCBs han women
- Le els o bo h dl-PCBs and ma ke PCBs we e posi i ely associa ed wi h age
- Indi iduals wi h lowe le el o educa ion had highe le els o PCDD/Fs han pa icipan s wi h uni e si y
quali ica ions
- Women who had b eas ed had signi ican ly lowe le els o PCBs han he es o all he pa icipan s
