# Impact of nonyphenol on soil fungi

Albert Kollmann, Isabelle Touton, Jacqueline Dubroca, Agathe Brault,
Christian Mougin

► To cite this version:

Albert Kollmann, Isabelle Touton, Jacqueline Dubroca, Agathe Brault, Christian Mougin. Impact of nonyphenol on soil fungi. 3rd European Meeting on Environmental Chemistry, Dec 2002, Genève, Switzerland. 2002. [hal-01605735](https://hal.science/hal-01605735)

HAL Id: hal-01605735

https://hal.science/hal-01605735

Submitted on 4 Jun 2020

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## 4-n-nonylphenol

- Numerous organic chemicals, including natural products and xenobiotics such as nonylphenol, are endocrine disrupters acting mostly as estrogen mimics,

- These chemicals are often detected in quantifiable levels in aquatic ecosystems, because of their intensive use for domestic, industrial and agricultural applications,

- Adverses effects have been noticed for a long time on larvae, embryos and adults forms of aquatic organisms: masculinization of females molluscs, feminization of male fishes, abnormalities of the reproduction in birds...

- Disrupters also accumulate in sewage sludge from urban origins, applied onto agricultural soils as amendments,

- The fate of these compounds are their impact on soil micro-organisms, higher plants and invertebrates are unknown,

- Filamentous fungi represent the highest biomass in the soil. They possess also receptors to sexual hormones, including steroids.

Our objectives are to assess the fate and the impact of nonylphenol (a mixture of isomers, NP, including 4-n-nonylphenol) on soil fungi, when the chemical is present in sludge used as a soil amendment.

## EXPOSURE OF FUNGI TO NONYLPHENOL

The soil from Versailles (a silt loam) was treated with NP. Other soil samples were mixed with sludges (95/5 ratio) from urban origins (Ambares, SA; Plaisir, SP), containing high levels of NP. Final NP amount in all samples was 40 mg/kg.

### Bioavailability of NP

Concentration of NP in the soil solution was determined after 4 hours.

It amounted to:
* **1.93 ± 0.23** mg L⁻¹ for the soil alone (8.8 10⁶ M)
* **1.46 ± 0.73** mg L⁻¹ for the soil/SA mixture (6.6 10⁶ M)
* **0.083 ± 0.004** mg L⁻¹ for the soil/SP mixture (3.8 10⁷ M)

>> lower bioavailability of NP in the presence of the sludge from Plaisir

$$ \text{4-nonylphenol Isotherms} \\ C_{\text{solid}} = 200 \text{ g/L} $$

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### Adsorption isotherms

>> 1 - NP adsorption on soil/sludges mixtures ≠ adsorption on soil
2 - adsorption capacity of SP > SA

### Half-lives of NP

4 days in the soil alone
16 days in soil/SA mixture
> 16 days in the soil/SP mixture

* Fusarium oxysporum (deuteromycete)
* Fusarium solani (ascomycete)
* Mucor racemosus (zygomycete)
* Phanerochaete chrysosporium (basidiomycete)
* Trametes versicolor (basidiomycete)

## ACUTE TOXICITY OF NONYLPHENOL

<table><thead><tr><th rowspan="2">Fungal strains</th><th colspan="4">IC<sub>50</sub> (M)</th><th rowspan="2">LC<sub>100</sub> (M)<br>Fungal<br>development</th></tr><tr><th>Germination of<br>mother spores</th><th>Fungal growth<br>after 2 days</th><th>Fungal growth<br>after 4 days</th><th>Germination of<br>daughter spores</th></tr></thead><tbody><tr><td>T. versicolor</td><td>/</td><td>> 10<sup>-3</sup></td><td>> 10<sup>-3</sup></td><td>/</td><td>> 10<sup>-3</sup></td></tr><tr><td>P. chrysosporium</td><td>7.5 10<sup>-6</sup></td><td>10<sup>-4</sup></td><td>5.0 10<sup>-4</sup></td><td>2.5 10<sup>-6</sup></td><td>> 10<sup>-3</sup></td></tr><tr><td>F. solani</td><td>4.1 10<sup>-4</sup></td><td>4.8 10<sup>-5</sup></td><td>> 10<sup>-4</sup></td><td>5.1 10<sup>-6</sup></td><td>> 10<sup>-3</sup></td></tr><tr><td>M. racemosus</td><td>10<sup>-4</sup></td><td>5.1 10<sup>-6</sup></td><td>5.3 10<sup>-5</sup></td><td>5.9 10<sup>-5</sup></td><td>10<sup>-4</sup></td></tr><tr><td>F. oxysporum</td><td>2.9 10<sup>-5</sup></td><td>5.0 10<sup>-6</sup></td><td>5.1 10<sup>-5</sup></td><td>5.1 10<sup>-6</sup></td><td>10<sup>-4</sup></td></tr></tbody></table>

IC, inhibiting concentration; LC lethal concentration

Five fungal strains belonging to the main taxonomic groups were selected. *T. versicolor* was unable to sporulate in laboratory conditions and was used as mycelial mats. Other strains were inoculated as spore suspensions. The impact of NP was examined as a dose-response relationship between chemical concentration and 2 endpoints: fungal growth (biomass production) and reproduction (germination rate of the spores).

>> 1 - sensitivity to NP depends on strains
2 - high sensitivity of mother spores of *P. chrysosporium*
3 - no dramatic effects at realistic concentrations in soil solution
4 - morphological defects at higher concentrations in *F. solani* and *F. oxysporum*

## INDUCTION OF ENZYMATIC ACTIVITIES

### CHRONIC TOXICITY OF NONYLPHENOL

Spores of fungi were inoculated in media, allowed to germinate, the resulting biomass was measured after 10 days, and the daughter spores were collected. That cycle was repeated 5 times, without NP, with 5 10⁻⁶ M NP or with decreasing concentration of chemical (2-fold dilution) at each inoculation.

#### 1 - fungal growth

no significant effect on *P. chrysosporium* and *M. racemosus*
slight stimulation on *F. solani* and *F. oxysporum* at the end of the treatment, without dose-response relationship

#### 2 - sporulation

no significant effect on *P. chrysosporium*
inhibiting effect on *F. solani* and *M. racemosus*, without dose-response relationship
strong stimulating effect on *F. oxysporum*

4-Days old cultures of *T. versicolor* were treated with NP. Laccase activity was measured after 3 days.

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## CONCLUSIONS

1 - the potential of NP to adversely impact fungi is low, because of a reduced exposure

2 - nevertheless, NP is toxic to fungi at high concentrations

3 - NP has been described in fungi to uncouple respiration and to affect the ATP-driven Ca⁺⁺ ion pumps

4 - Does the chemical also affect sexual hormone pathways in filamentous fungi?

>> 1 - NP is an inducer of laccases in *T. versicolor* cultures

2 - the 4-n-isomer is more potent that the mixture of isomers to stimulate laccase production

3 - NP also induces peroxidase activities in *P. chrysosporium* (not shown)

Pojer K., Davoine P., Dubroca J., Dur J.-C., Mougin C., Jolivalt C., Cravedi, J.-P. and Chaplain V. 1999. Behaviour and fate of nonylphenol in soils containing contaminated sewage sludge. Biodépoll'99 : Biotechnology in environmental protection, Rennes

Dubroca J., Brault A., Kollmann A., Touton I., Jolivalt C., Kerhoas L. and Mougin C. 2002. Biotransformation of nonyphenol surfactants in soils amended with contaminated sewage sludges. Dans Environmental Chemistry. Lichtfouse E., Dudd S., Robert D. eds, Springer (soumis).

Mougin C., Kollmann A. and Jolivalt C. 2002. Enhanced production of laccases by the fungus *Trametes versicolor* by the addition of xenobiotics. Biotechnology Letters, 24:139-142.

Karney A.J., Powell S.I. and Davies J.M. 1997. Effect of nonylphenol on growth of Neurospora crassa and Candida albicans. Applied and Environmental Microbiology, 63:1312-1317.