# Biotransformation of nonylphenol surfactants in soils amended with contaminated sewage sludge

Christian Mougin, Jacqueline Dubroca, Agathe Brault, Albert Kollmann, Isabelle Touton, Claude Jolivalt, Lucien Kerhoas, Veronique Chaplain

► To cite this version:

Christian Mougin, Jacqueline Dubroca, Agathe Brault, Albert Kollmann, Isabelle Touton, et al. Biotransformation of nonylphenol surfactants in soils amended with contaminated sewage sludge. 2nd European Meeting on Environmental Chemistry, Dec 2001, Dijon, France., 2001. hal-01600091

HAL Id: hal-01600091

https://hal.science/hal-01600091

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<figure><img src="image_1.png" /></figure>

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# Biotransformation of nonylphenol surfactants in soils amended with contaminated sewage sludges

C. Mougin, J. Dubroca, A. Brault, A. Kollmann, I. Touton,

C. Jolivalt, L. Kerhoas and V. Chaplain

Unité de Phytopharmacie et Médiateurs Chimiques

Route de Saint-Cyr, F-78026 Versailles Cedex

email: mougin@versailles.inra.fr
Alkylphenol ethoxylates (APEs)

* nonionic surfactants used in domestic, industrial and agricultural applications

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

$n = 1 \text{ to } 18 \rightarrow \text{APEs}$

$n = 0 \rightarrow \text{nonylphenol NP}$

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

linear chain: 4-n-NP

branched nonyl groups: mixture of 18 isomers

$S_{\text{wat}} (\text{mg/L})$ 7.0

5000.0

$\text{Log P}$ 5.76

5.92

$\text{Henry's C (atm/M)}$ 3.4 $10^{-5}$

0.43 $10^{-5}$

* enter the environment through wastewater and sewage sludge
Environmental hazard of NPs

* 80% of total APEs

US production: 225,000 T/year

* toxic effects on wildlife and humans

multiple physiological effects due to uncoupled respiration

endocrine disruptor

proliferator of breast cancer in women

* low amounts of discharged NPs in water: < 10 µg/L

acute and chronic toxicity to aquatic organisms well known

* high amounts in sewage sludge: 1 g/kg

fate of NPs in soils enriched with sludges?

effects on soil (micro)-organisms?
# Objectives

1) to assess the ecotoxicological risk induced from soil amendment with wastes

2) to estimate exposure of soil (micro)-organisms to NPs contained in sewage sludge from urban origin

## Topics:

### Biotransformation of NPs:

* in an agricultural soil

* in soil/sludge mixtures

* by filamentous fungi

* by enzymes purified from fungi
Biotransformation of NPs in an agricultural soil

* 27 g dry silt loam sieved at 2 mm, 40 mg/kg isoNPs*, 80% MHC, 25°C, darkness

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

¹⁴C balance +
HPLC analysis

→ T½ ≈ 4 days
Biotransformation of NPs in soil/sludge mixtures

* 25 g dry silt loam + 2 g dry sludge, 40 mg/kg NPs*, 80% MHC, 25°C, darkness

<table><thead><tr><th>anaerobic digested sludge</th><th>Ambares (33)</th><th>Plaisir (78)</th></tr></thead><tbody><tr><td>pH</td><td>7.8</td><td>12.5</td></tr><tr><td>NP content (mg/kg dw)</td><td>200.0</td><td>506.00</td></tr><tr><td>PAH content /</td><td>0.51</td><td>0.43</td></tr><tr><td>PCB content /</td><td>0.089</td><td>0.071</td></tr><tr><td>Metal ions /</td><td>8880.71</td><td>113.86</td></tr></tbody></table>

* After 96 days

<table><tbody><tr><td>non extractable <sup>14</sup>C</td><td>↓</td><td rowspan="3" style="vertical-align: middle;">a 8-day lag phase, then</td></tr><tr><td>extractable <sup>14</sup>C</td><td>↓</td></tr><tr><td><sup>14</sup>C mineralized</td><td>X 2</td></tr><tr><td>→ T<sup>1</sup>&frasl;<sub>2</sub> NPs (days)</td><td>16</td><td>&gt; 16</td></tr></tbody></table>

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

INRA

Institut National de la Recherche Agronomique
Biotransformation of NPs by filamentous fungi

* 10-mL liquid cultures, 1.1 mg/L NPs after 3 days of growth, 25°C

<table><thead><tr><th rowspan="2"></th><th rowspan="2">Strains</th><th colspan="2">T<sup>1/2</sup> NPs (days)</th></tr><tr><th>4nNP</th><th>isoNPs</th></tr></thead><tbody><tr><td rowspan="4">Laboratory</td><td>Trametes versicolor</td><td>1.0</td><td>&lt; 1.0</td></tr><tr><td>Phanerochaete chrysosporium</td><td>6.0</td><td>3.0</td></tr><tr><td>Cunninghamella elegans</td><td>1.0</td><td>2.0</td></tr><tr><td>Cunninghamella echinulata</td><td>1.0</td><td>2.0</td></tr><tr><td rowspan="3">Ambares</td><td>Mucor racemosus</td><td>2.0</td><td>3.0</td></tr><tr><td>Fusarium oxysporum and solani</td><td>2.0</td><td>&gt; 8.0</td></tr><tr><td>Mucor hiemalis</td><td>1.5</td><td>5.0</td></tr><tr><td rowspan="3">Plaisir</td><td>Fusarium solani</td><td>1.0</td><td>&gt; 8.0</td></tr><tr><td>Fusarium sp.</td><td>1.5</td><td>&gt; 8.0</td></tr><tr><td>Fusarium oxysporum</td><td>2.0</td><td>6.0</td></tr></tbody></table>
# Biotransformation of NP by laccases purified from *T. versicolor*

<figure><img src="image_6.png" /><figcaption>* 5 mg/L 4nNP, 1 U laccase, 30°C
Kinetics at pH 4</figcaption></figure>

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

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

**Analysis** : HPLC 50 % CH₃CN
column C8, gradient CH₃CN/H₂O, 279 nm
limit of detection: 7 µg/L
Metabolites formed from NP by purified laccases

<figure><img src="image_9.png" /></figure>
Conclusions and perspectives

* NPs are rapidly transformed in soil by mineralization and stabilization (bound residues)

* the biodegradation rate is somewhat decreased in soil/sludge mixtures sludge toxicity (organic compounds and metal ions) high biological oxygen demand (high amount of organic matter) increased NP adsorption (Kd: 45 S, 59 S/A, 1084 S/P)

→ as a consequence, exposure of soil organisms to NPs may be increased

* fungi isolated from soil/sludge mixtures are ± efficient in transforming NPs

* fungal laccases are involved in NP polymerization one of the mechanisms responsible for bound residue formation

→ it remains important to assess the effects of NPs on soil (micro)-organisms