Herbicides and nonylphenol in sewage sludge:
occurrence and ecotoxicological impacts

Aline Ghanem, Jacqueline Dubroca, Philippe Bados, Luiz Felippe de Alenastro, C. Staempfli, Virginie Huteau, Agathe Brault, Veronique Chaplain, Christian Mougin

► To cite this version:

Aline Ghanem, Jacqueline Dubroca, Philippe Bados, Luiz Felippe de Alenastro, C. Staempfli, et al.. Herbicides and nonylphenol in sewage sludge: occurrence and ecotoxicological impacts. SETAC Europe 16th Annual Meeting, May 2006, La Haye, Netherlands. 2006. [hal-01600752](https://hal.science/hal-01600752)

HAL Id: hal-01600752

https://hal.science/hal-01600752

Submitted on 3 Jun 2020

HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers.

L'archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d'enseignement et de
recherche français ou étrangers, des laboratoires
publics ou privés.

<figure><img src="image_1.png" /></figure>
# Herbicides and nonylphenol in sewage sludge:
occurrence and ecotoxicological impacts

Ghanem A., Dubroca J., Bados P., de Alencastro F., Staempfli C., Huteau V., Brault A., Chaplain V. and Mougin C.

mougin@versailles.inra.fr

http://www-pessac.versailles.inra.fr

<figure><img src="image_2.png" /></figure>

Phytopharmacie
Versailles
Xénobiotiques
Toulouse

<figure><img src="image_3.png" /></figure>

CECOTOX
Lausanne

<figure><img src="image_4.png" /></figure>

Santé Publique et
Environnement
Chatenay-Malabry

<figure><img src="image_5.png" /></figure>

Biochimie
Paris

<figure><img src="image_6.png" /></figure>

Gestion Biologique
des Sols
Angers
# Rationale

## Scientific

* **Selection of sludge and contaminants**
  type of process and sludge treatment in WWTP
  persistent organic pollutants ?
  analytical methods
  variation of concentration in sludge with time and WWTP

* **Transfer of contaminants**
  amounts and forms
  waters, plants, webfoods

* **Impact of contaminated sludge**
  soil properties and microbial activity
  écotoxicology
  relationships environment / health, EDCs

## Socio-economic

* **Management and sustainability of sludge spreading onto agricultural soils**
# Objectives of our research program

* Determine the occurrence of selected pesticides in sludge samples

- development of analytical methods

- annual monitoring in various sludge samples

* Identify possible ecotoxicological effects of the pesticides when applied onto the soils in contaminated sludge

- fate of the pesticides in the soil

- ecotoxicity on selected organisms
# The chemicals

## Atrazine

<figure><img src="image_7.png" /></figure>

<figure><img src="image_8.png" /></figure>

### DEA

<figure><img src="image_9.png" /></figure>

et

### DIA

<figure><img src="image_10.png" /></figure>

## Diuron

<figure><img src="image_11.png" /></figure>

<figure><img src="image_12.png" /></figure>

### DCPMU

<figure><img src="image_13.png" /></figure>

## Glyphosate

<figure><img src="image_14.png" /></figure>

<figure><img src="image_15.png" /></figure>

### AMPA

<figure><img src="image_16.png" /></figure>

## Nonylphenol

<figure><img src="image_17.png" /></figure>
# 1 – Sludge samples contain pesticides

≈ 80 samples from 4 WWTPs during 1 year near Versailles (France)

<table><thead><tr><th>Compound</th><th>Dried</th><th>Composted</th><th>Limed 1</th><th>Limed 2</th></tr></thead><tbody><tr><td>Atrazine + met</td><td>/</td><td>/</td><td>/</td><td>/</td></tr><tr><td>Diuron ●</td><td>11.2</td><td>26.5</td><td>20.0</td><td>46.6</td></tr><tr><td>DCPMU</td><td>/</td><td>/</td><td>/</td><td>/</td></tr><tr><td>Glyphosate ●</td><td>1.1</td><td>1.0</td><td>1.4</td><td>0.4</td></tr><tr><td>AMPA ●</td><td>20.3</td><td>12.4</td><td>11.5</td><td>2.8</td></tr><tr><td>Nonylphenol ●</td><td>61.7</td><td>132.8</td><td>90.9</td><td>62.5</td></tr></tbody></table>

• ppb (µg/kg dry sludge) / • ppm (mg/kg)

French regulation: PCB 0.8 and BaP 2 ppm / EC regulation: nonyphenol 50 ppm
► It is necessary to study their fate in the soil

Light 16 h, 21°C / darkness 8 h, 18°C

<figure><img src="image_18.png" /></figure>

Transformation

• Mixture of labelled Gly/Diu/NP at 6/17/7 ppm

• Soil

• Soil + spiked sludge at 30 and 6 T/ha

► mineralised / extractable / stabilised metabolic maps

Mobility

• Transfer soil / higher plant

<figure><img src="image_19.png" /></figure>

Water contamination
2 – The mineralisation of the pesticides is decreased

<figure><img src="image_20.png" /></figure>

• Composted sludge has contrasted effects at high dose

• Limed sludge induces a 7-day lag phase at high dose
3 – The persistence of the pesticides is increased

<figure><img src="image_21.png" /></figure>

<figure><img src="image_22.png" /></figure>

With time
Mineralisation increased
Extractability decreased
Bound residue increased

<figure><img src="image_23.png" /></figure>

With sludge at 30 T/ha
* Mineralisation decreased
* Extractability maintained

<figure><img src="image_24.png" /></figure>

With sludge at 6 T/ha
* Intermediate situation
# 4 – Leachates contain traces of pesticides

Less than 1 % of initial radioactivity with a rainfall of 20 mm

<table><thead><tr><th>Conditions</th><th>D + 45<br>µg/L</th><th>D + 60<br>µg/L</th><th>D + 90<br>µg/L</th></tr></thead><tbody><tr><td>Soil alone</td><td>2.1</td><td>1.6</td><td>1.2</td></tr><tr><td>+ Dried sludge (30 T/ha)</td><td>2.5</td><td>0.9</td><td>1.2</td></tr><tr><td>+ Dried sludge (6 T/ha)</td><td>0.5</td><td>0.3</td><td>0.2</td></tr><tr><td>Soil alone</td><td>14.5</td><td>16.2</td><td>11.0</td></tr><tr><td>+ Composted sludge (30 T/ha)</td><td>14.9</td><td>12.5</td><td>15.2</td></tr><tr><td>+ Composted sludge (6 T/ha)</td><td>4.8</td><td>5.2</td><td>4.7</td></tr><tr><td>Soil alone</td><td>14.5</td><td>16.2</td><td>11.0</td></tr><tr><td>+ Limed sludge (30 T/ha)</td><td>29.6</td><td>25.2</td><td>25.4</td></tr><tr><td>+ Limed sludge (6 T/ha)</td><td>3.4</td><td>3.8</td><td>3.7</td></tr></tbody></table>

* Nonylphenol is the main compound detected using HPLC
# 5 – Sludge modify the transfer of pesticides from the soil to higher plants

Less than 0.05 % of the radioactivity during the period of growth

<table><thead><tr><th>Conditions</th><th>Radish (15 days)<br>µg/g</th><th>Wheat (45 days)<br>µg/g</th></tr></thead><tbody><tr><td>Soil alone</td><td>/</td><td>0.9</td></tr><tr><td>+ Dried sludge (30 T/ha)</td><td>/</td><td>0.07</td></tr><tr><td>+ Dried sludge (6 T/ha)</td><td>/</td><td>/</td></tr><tr><td>Soil alone</td><td>6.0</td><td>0.7</td></tr><tr><td>+ Composted sludge (30 T/ha)</td><td>1.1</td><td>1.2</td></tr><tr><td>+ Composted sludge (6 T/ha)</td><td>1.8</td><td>1.6</td></tr><tr><td>Soil alone</td><td>6.0</td><td>0.7</td></tr><tr><td>+ Limed sludge (30 T/ha)</td><td>4.3</td><td>0.1</td></tr><tr><td>+ Limed sludge (6 T/ha)</td><td>/</td><td>1.1</td></tr></tbody></table>
It is necessary to study their ecotoxicological impacts.

<figure><img src="image_25.png" /></figure>

## Assays including 3 ecological levels

* **Degraders**

microbial functional diversity
« litter bag »
structural diversity
PLFA

* **Producers**

germination (%, delay) of plants
biomass

* **Consumers**

mortality, reproduction of springtails

<figure><img src="image_26.png" /></figure>

Estrogenicity assessment

human cell lines MELN

<figure><img src="image_27.png" /></figure>

Risk for humans
6 – Pesticides have no clear impact on the litter-bag

<figure><img src="image_28.png" /></figure>

* Dried sludge increases the soil biological activity at low dose

* Composted sludge decreases the soil activity at high dose
# 7 – The fungi/bacteria ratio is increased

<figure><img src="image_29.png" /></figure>

* Sludge intake generally increases the fungi/bacteria ratio

* The highest effects are noticed with the dried sludge at low dose

* There is no clear effect of pesticides

<figure><img src="image_30.png" /></figure>

* Dried sludge at high dose induces the development of fungi at the soil surface
## 8 – Sludge are phytotoxic on wheat at high doses

<figure><img src="image_31.png" /></figure>

% of germination, delay, biomass production

* Sludge are fertilizers at low dose

* Dried sludge inhibits plant development at high dose (fungal proliferation)

<figure><img src="image_32.png" /></figure>

* Spiked sludges are phytotoxic at high dose

* Phytotoxicity could be due to pesticides
9 – Sludge are toxic for springtails

<figure><img src="image_33.png" /><figcaption>F. candida</figcaption></figure>

<figure><img src="image_34.png" /></figure>

* No effect at low dose

* Dried sludge inhibits sprintails life at high dose (fungal proliferation)

* Limed sludge inhibits sprintails survival at high dose (pH?)

<figure><img src="image_35.png" /></figure>

* Limed sludge strongly inhibits springtail survival at high dose

* Toxicity could be due to pesticides
# 10 – Leachates induce no clear estrogenic effects

<figure><img src="image_36.png" /></figure>

<figure><img src="image_37.png" /></figure>

<figure><img src="image_38.png" /><figcaption>MELN cells</figcaption></figure>

* Dried sludge only induced significant estrogenic effects on cells at high dose

* That effect seems strengthened by pesticides (nonylphenol ?)
Conclusions and perspectives

* Urban sewage sludge contain pesticides at relatively high concentrations

* The persistence of pesticides entering the soil through sludge is increased

* Eco-toxic effects have been evidenced at the high dose of sludge (worst case), and not in an agronomic reality

* The effects of sludge are in general higher than these of added pesticides

* The process in WWTPs must be improved to degrade organic pollutants
# Acknowledgements

* The colleagues of the 4 research institutes
* scientists, research assistants, technicians, administrative staff

* The students
* PhD (Aline Ghanem) and others

* The workers of the WWTPs and water treatment companies

* The French Agency for Environment and Energy Management
* (agreement 0275048)

ADEME

<figure><img src="image_39.png" /></figure>