1. Emerging contaminants and endocrine disruptors in Brazil: an overview
4th Latin American and Caribbean regional meeting on the Strategic Approach to International Chemicals Management (SAICM), Mexico City, Mexico, August 2013
Emerging contaminants and endocrine disruptors in Brazil: an overview
Wilson F. Jardim and co-workers
Universidade Estadual de Campinas – UNICAMP
São Paulo, Brazil

2. Previous and current scenario:
Poor sanitation conditions;
Very limited data on the presence of emerging contaminants (including EDC) in natural waters, including drinking waters;
Starting in 2005 with a detailed investigation on both source and drinking waters in the State of São Paulo;
16 compounds were selected to the analyzed based upon estrogenicity, frequency of occurrence and concentration;
Using a BLYES assay (luminescence) to compare the biological response to the chemical burden found in the water samples;
Choosing appropriate compound that could be used as indicator of estrogenicity;
In 2010 a national survey on the presence of caffeine in drinking water was carried out in 19 capitals;
In 2012 the survey was repeated with 23 capitals (~ 40 million people);
In vivo studies started in 2013;
25 pesticides have just being included in the list of investigated compounds;

3. Compounds investigated in the period 2010-2013 in drinking water
Endogenous Hormones E1 E2 E3
PROG
TTN
Synthetics Hormones
EE2
MEE
NGT
DES
Xenostrogens
ATZ
nOP
nNP
BPA
TCS
PhPh
CAF

4. Analytical Tools: LC-MS/MS YEAST BIOASSAY

5. Human estrogen receptor
Sample with EDC
Human estrogen receptor
hER gene
Núcleo
GPD
ADH1
ERE
ADH1
luxC
GPD
luxE
BLYES
luxD
luxA
luxB
frp
LD – 0,09 ±0,02 ng E2/L
Luminescence is
measured

6. Pippeting of samples
Pippeting of cultures
Overnight incubation of strains (30 °C)
Bioluminescence reading

7. BLYES for the 5 target compounds

8. Expression of results Chemical analysis ng/L for each compound BLYES
ng-Equivalent of
E2/L
EACH COMPOUND WERE TESTED IN THE BLYES, SO IT IS POSSIBLE
TO COMPARE TO MULTIPLY ITS POTENCY BY ITS CONCENTRATION
AND VERIFY ITS CONTRIBUTION TO THE TOTAL POTENCY OF THE SAMPLE
EXPECTED POTENCY X OBSERVED POTENCY

9. QUATITATIVE COMPARISION BLYES more sensitive
no agreement…
Expected
Observed 10

10. Estrogenicity varies along the year dry season x wet season

11. IN VIVO BIOASSAYS

12. MATURE AND IMATURE FEMALES EXPOSED TO DRINKING WATER EXTRACTS
Protocolo A – OECD 440
Protocolo B – EPA
UTEROTHROPHIC ASSAY
21 DAYS OF LIFE
SEXUAL MATURATION
21 TO 42 DAYS OF LIFE
GENETIC, PHYSICAL AND MORFOLOGICAL ALTERATIONS IN:
NO RESULTS, YET…..

13. CAMPINAS - PREVIOUS RESULTS 2010-2012
Drinking Water
Surface Water
14/77
15/77
48/77
28/77
31/77
70/77
0/49
1/49
4/49
24/49
47/49

14. Estrogenic activity (BLYES)
Blyes observed response (ngE2 equiv L-1)
Caffeine (ng L-1)

15. PCA
Samples : 103
Outliers: 1

16. BRAZILIAN NATIONAL
CAFFEINE SCREENING
SOURCE AND
DRINKING WATER

17. BRAZILIAN REGIONS North Region Northeast Region Central Region
Southeast Region
South
Region

18. caffeine screening
SOUTH REGION
Drinking water ng L-1
Source water ng L-1
2600
1700
2700
1300 20 25
165

19. caffeine screening
SOUTHEAST REGION
Drinking water ng L-1
Source water ng L-1
19000 40
250
1000
200 30
270
160 10
600

20. 4
190 3 7 35
100 5 8 75
Drinking water ng L-1
caffeine screening
NORTHEAST REGION

21. caffeine screening
NORTH REGION 10 35 55
130
Drinking water ng L-1

22. caffeine screening
CENTRAL REGION 10
600
1800 5 55 60 20
Drinking water ng L-1

23. Conclusions
Caffeine is a resilient indicator of both treated and untreated sewage inputs and correlates not only to fecal coliforms, but also to estrogenic activity;
Routine monitoring of some selected EC in both surface and drinking waters (proposed during the revision of the Brazilian Drinking Water Act);
Just published the first attempt to regulate Triclosan to protect aquatic organisms in natural waters;
Urgent need for biological assays to evaluate estrogenic activity both in vitro and in vivo;
Which EDCs are the most important to monitor?
Synergy and antagonism in real scale.

24. wfjardim@iqm.unicamp.br http://lqa.iqm.unicamp.br
4th Latin American and Caribbean regional meeting on the Strategic Approach to International Chemicals Management (SAICM), Mexico City, Mexico, August 2013
Thank you!