# Accumulation and chronic effects of cobalt on the gastropod Radix balthica

Maëva Marimoutou, Gilles Bareille, Mathieu Milhe-Poutingon, Patrick

Baldoni-Andrey, Amiel Boullemant, Cecile Courreges, Clémentine Gelber,

Sandra Mounicou, Hélène Tabouret, Séverine Le Faucheur

► To cite this version:

Maëva Marimoutou, Gilles Bareille, Mathieu Milhe-Poutingon, Patrick Baldoni-Andrey, Amiel Boullemant, et al.. Accumulation and chronic effects of cobalt on the gastropod Radix balthica. SETAC Europe 32nd annual meeting, May 2022, Cophenague, Denmark. hal-03960403

HAL Id: hal-03960403

https://hal.science/hal-03960403

Submitted on 27 Jan 2023

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Distributed under a Creative Commons Attribution 4.0 International License
# Accumulation and chronic effects of cobalt on the gastropod Radix balthica

Maëva Marimoutou¹, G. Bareille¹, M. Milhe-Poutingon¹, P. Baldoni-Andrey², A. Boullemant³, C. Courrèges¹, C. Gelber², S. Mounicou¹, H. Tabouret¹, S. Le Faucheur¹

¹Université de Pau et des Pays de l'Adour, E2S-UPPA, CNRS, IPREM, Chaire Ecotox, Pau, France; ²TotalEnergies, Pole d'Études et de Recherche de Lacq, France; ³Rio Tinto Closure, France (Email: maeva.marimoutou@univ-pau.fr)

## INTRODUCTION

Gastropods are widespread mollusks living at the bottom and in shallow water of freshwater systems. First consumers of periphyton and plants, they represent an important source of food for large crustacean and fish. Toxic effects due to contamination such as an increase in metal would thus impede food sustainability of ecosystems. Gastropod shells have the interesting feature of growing throughout their life by integrating some of the chemical elements present in their environment, including metals.

<figure><img src="image_2.png" /><figcaption>Shell formation (1) (2)</figcaption></figure>

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

In this study, cobalt (Co) is the metal of interest due to its increasing use in recent technologies of transport or energy storage(3). Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICPMS - micrometer scale) was used to explore the variability of metals in shells with two objectives :

1) To study Co effects towards gastropod growth and reproduction during 28 days in outdoor mesocosms

2) To examine the relationships between ambient Co concentrations and Co accumulation in shell gastropods

## MATERIALS AND METHODS

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

### Exposure conditions

* 28 days in June 2021

* 4 studied Co concentrations : 0.7 ± 0.2 µg.L⁻¹ (control), 2.4 ± 0.8 µg.L⁻¹, 24.1 ± 0.8 µg.L⁻¹ and 56 ± 17 µg.L⁻¹

* 3 boxes of 32 organisms of *Radix balthica* collected *in situ* placed in each artificial stream

* Sampling at D0, D7, D14, D21 and D28

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

©TotalEnergies

### Once a week

* Physicochemical parameters : Conductivity, pH, oxygen and turbidity

* Water analysis : Anion, cation and dissolved organic matter concentrations

* Organism analysis : Shell measurements (length and width) and number of egg masses

* Shell analysis : Microchemical analyses of metals in the shells performed using LA-ICP-MS

### LA-ICP-MS analysis

<figure><img src="image_6.png" /><figcaption>Day 0</figcaption></figure>

<figure><img src="image_7.png" /><figcaption>5.3 ± 0.3 mm</figcaption></figure>

<figure><img src="image_8.png" /><figcaption>+ 1 mL H₂O₂ (2%) during 12h</figcaption></figure>

<caption>8.1 ± 0.5 mm</caption>

<figure><img src="image_9.png" /><figcaption>Shell cleaning</figcaption></figure>

<figure><img src="image_10.png" /><figcaption>Shell measurements</figcaption></figure>

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

<figure><img src="image_12.png" /><figcaption>Shell preparation</figcaption></figure>

<figure><img src="image_13.png" /><figcaption>External surface of the end of shell (peristome)
3 replicates per conditions and each sampling day</figcaption></figure>

<figure><img src="image_14.png" /><figcaption>LA-ICP-MS analysis</figcaption></figure>

<figure><img src="image_15.png" /><figcaption>IR 1030 nm femtosecond laser ablation coupled to an Agilent 8900 ICP-MS Triple Quad</figcaption></figure>

## RESULTS

### 1) Co effects towards gastropods

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

* Mortality increased significatively at Day 28 (p<0.05)

* No significant mortality induced by Co exposure concentration

* No effect of Co concentrations was found on the shell length (p>0.05)

* Similar results were obtained for the width (p>0.05; data not shown)

* Growth in length and width only between day 0 and day 7

<figure><img src="image_17.png" /></figure>

* Maximum number of egg masses at Day 14 in the control (0.7 µg.L⁻¹)

* At Day 21 and 28: number of masses increased with Co concentration

* Number of neonates decreased with Co concentration

* Probably due to delay in the hatching time

### 2) Cobalt distribution in shell

#### A) Co distribution in shell by mass-spectrometry imaging at Day 28

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

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

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

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

* In control, Co distributed evenly in the shell at very low concentration (<1µg/g)

* At all exposure concentrations, higher Co concentration mainly found on the shell external surface

* At the 24.1 and 56 µg/L, shell Co concentrations were high at both internal and external surfaces

#### B) Co concentrations in external shell surface as a function of exposure time

<figure><img src="image_22.png" /><figcaption>External surface : recent part</figcaption></figure>

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

<figure><img src="image_24.png" /><figcaption>Analysis within a single growth ridge</figcaption></figure>

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

* Significant increase in shell Co concentration was observed from 2.4 µg/L Co at Day 7

No further increase was observed at longer exposure time

* Shell Co concentration further increased at 24.1 µg/L. A plateau was measured at 56 µg/L Co

## CONCLUSION

* The present study demonstrates the impact of Co on the reproduction of *R. Balthica*. A delay in hatching was observed in Co-exposed gastropods. This delay is dependent of exposure Co concentration with a minimum effect at higher Co concentrations. The number of neonates is much lower at the higher Co concentrations.

* Co did not show a negative impact on the shell growth at the micrometer level.

* Co mainly distributes in the external shell surfaces. Shell Co accumulation was already measured at the end of the first week of exposure and no further increase was observed with longer exposure time. Future analysis of Co accumulation in the gastropod soft bodies as a function of time will allow us to better understand Co pathway from water to the shell and to assess the potential use of gastropod shells as bioindicator of metal contamination.

## ACKNOWLEDGEMENT

We are grateful to TotalEnergies for allowing us to perform our experiments on their site. All

members of Chair Ecotox and TotalEnergies helped with field preparation and sampling, especially

S.Sebihi, J.M Dalens, F. Mounede, C. Pillitteri.

## REFERENCES

1. Checa AG, Frontiers in Marine Science, 2018

2. Marin F, Le Roy N, Benjamin M, Frontiers in Bioscience, 2012

3. IFP Energies nouvelles, 2020

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

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

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