New protocols for the selection and rearing of Metoncholaimus pristiurus and the first evaluation of oxidative stress biomarkers in meiobenthic nematodes

https://doi.org/10.1016/j.envpol.2020.114529Get rights and content

Highlights

  • Endoskeleton of Sepia officinalis was efficient to select Metoncholaimus pristiurus.

  • Porcellio scaber powder was sufficient to feed M. pristiurus in the laboratory.

  • Oxidative stress biomarkers were studied for the first time in meiobenthic nematodes.

  • M. pristiurus could be one of the first taxa responding to zinc and/or permethrin.

Abstract

Meiobenthic nematodes have been designated as sensitive global models in the development of biomonitoring and ecotoxicology monitoring programs howbeit the sensitivity of these organisms against oxidative stress biomarkers have never been addressed. The present study aimed to decipher this research axis after selecting and culturing a single nematode species from an entire community through original laboratory protocols. The purpose of this investigation was to change the grain size of the sediment into the immediate environment of nematodes by progressively adding a biosubstrate made from Sepia officinalis endoskeletton. At the end of the experiment, Metoncholaimus pristiurus became the unique component of the nematode species when the sediment was enriched with 80% of S. officinalis powder. After the mono-species level had been achieved, the selected species was fed on an another biosubstrate made from bodies of Porcellio scaber under the identical laboratory controlled conditions of light and temperature adopted during the selection process. Accordingly, the bioassay protocol this study layed new foundations for the study of meiobenthic nematodes in the biomarker field. Our results revealed that, in case of M. pritiurus, discernible oxidative stress responses are valid for catalase and gluthatione S-transferase. Indeed, for both enzymes, a clear increase in the activity was recorded, and the response was more reinforced when zinc and permethrin were administrated in combination. The relevance of the protocols proposed in this work parallels their global applicability to reach and maintain the monospecific level in laboratory by using biosubstrates made from animals widely distributed. It is true also that our data provided the first results in terms of biochemical biomarkers for meiobenthic nematodes and showed that the selected taxa, M. pristiurus, could be one of the first marine taxa responding early to the tested stressors, zinc and permethrin, even at very low concentrations.

Introduction

In many Mediterranean countries, pesticides are still used for disease control in agriculture (Daaboub et al., 2008). Several surveys have indicated that such activities lead to widespread contamination on a local, regional, and global scale. Thus, alarming levels of pesticides remain present in the environment, as well as in human beings (Khazri et al., 2017). One of the most widely used pesticides is permethrin (C21H20Cl2O3), which is an insecticide from the pyrethroid family. It is widely considered as a control for insects such as mosquitoes, cabbage worms, corn earworms, potato beetles, etc. (Wang et al., 2016). Its broad spectrum of activity explains its wide range of use in the processing of various crops such as tobacco, cereals, potatoes, sunflowers, and mushrooms (Wang et al., 2016). In addition to acting as an agricultural pesticide, it is also used for domestic treatment of military clothing and mosquito nets. The presence of this insecticide in aquatic environments has shown its major impact on both pelagos and benthos; it is especially characterized by its non-polarity and high adsorption to sediment particles (Woudneh and Oros, 2006).

More problems are expected to be experienced with permethrin in aquatic environments since zinc is commonly used in the manufacturing of phytosanitary products like insecticides (Rodier, 1996; Sarkar, 2002). Zinc is known to be one of the most abundant metals in the Earth’s crust, ranging from 0.9 mg/kg to 169 mg/kg (Roney, 2005). It is naturally present in the environment, mainly in the form of zinc sulfide. Many other industries use a lot of zinc, especially metallurgy in the production of alloys. It is also used for the galvanization of metal parts, in the manufacturing of pigments for dyeing, in varnish, and as rat poison (Rodier, 1996; Sarkar, 2002).

Species-specific responses to pollutant exposure are the basis of most experimental studies in ecotoxicology (Hägerbäumer et al., 2015). Dealing with these problems is a huge and complex task. It requires developing and testing simple, inexpensive, early, and accurate ecotoxicological methods for monitoring and assessment. Such methods could be based on the use of biochemical biomarkers that can work experimentally at a reasonable cost. Hence, these tools provide precious and early information, thereby linking the health of an organism and its exposure to pollutants.

Free-living marine nematodes are known to be well suited as bioindicators for monitoring studies in marine environments and bioassays because only small sediment samples are needed, which does not require preliminary sieving in the field (Moreno et al., 2011). These worms are ubiquitous and occupy an important link in the food chain, feeding on microalgae and bacteria and, in turn, being preyed upon by macrobenthic predators such as polychaetes, crabs, and fishes (Albertini-Berhaut, 1974; Fitzhugh and Fleeger, 1985; Rivain, 1984). They are expected to be highly susceptible to sediment-associated pollutants because they live and feed on the sediment (Semprucci et al., 2018). So, any effects of permethrin and/or zinc are likely to be detected sufficiently early using nematodes before passing through the whole system and then onto humans.

Here, we consider taxonomic tools to be generally too complex to be used and presume that we must apply other tools, principally molecular and cellular, for the early detection of environmental risks (Boufahja et al., 2015; Boufahja et al., 2016). In fact, the biomarker approach is widely used in environmental biomonitoring, using macrofauna such as mussels and polychaetes (Freitas et al., 2015), but never for meiobenthic nematodes. Such an approach, if we can apply it to these worms, will allow us to identify interactions between organisms and contaminants and detect sublethal effects earlier than when using the bio-indication approach (Sarkar, 2002).

The main goals of this study were to (1) select one nematode species from an entire community of meiobenthic nematodes, (2) test the efficiency of some nutritive substrates in the rearing of the selected species, and (3) emphasize the importance of using various biochemical biomarkers in monitoring the concentration of permethrin and zinc in sediment, either separately or combined.

Section snippets

Sediment collection and measurements

Meiobenthic organisms were collected on January 1, 2015 (N0) (0700 h UTC) by gently pushing plexiglass cores (3.6 cm inner diameter) into the sediment at a depth of 10 cm at the ‘Career Bay’ site (37° 14.043′ N, 9° 50.162′ E) located in the Channel zone of Bizerte lagoon (Tunisia). This was the same site as Boufahja et al. (2016) previously considered but this time, samples were taken from the infralittoral zone and not from the intertidal zone. ‘Career Bay’ was also chosen for further study,

Abiotic factors

On day N0, abiotic factors at the site ‘Career bay’ were recorded as follows: depth, 0.3 m; salinity, 37.1 PSU; temperature, 7.94 °C; dissolved oxygen, 12.7 mg L−1; and pH, 8.52. The sediments collected were composed of 53.82% ± 1.13% silt/clay and contained 10.51% ± 0.54% organic matter and 28.20% ± 0.63% water. The coarse fraction had a mean grain size equal to 0.58 ± 0.31 mm.

Meiobenthic community at ‘Career bay’

The meiobenthic community collected on day N0 from ‘Career bay’ was characterized by an average abundance of

Discussion

For the purposes of environmental impact assessments for polycyclic aromatic hydrocarbons and heavy metals, meiofauna are generally more tolerant of the toxicity of these compounds than other more vulnerable taxa, which may be more efficient indicators of other less toxic types of stressors, especially those related to seawater. Within this context, our research clearly adds to a large and growing body of evidence showing that meiobenthic nematodes could be valid and early predictors of the

Conclusions

It is likely that all of the explanations above contributed partly to the significant relationships between diversity and enrichment with S. officinalis in our experiment. Both meiofauna in general and the nematode community used in this study may be relatively sensitive to the disturbance we applied, in part because of microhabitat homogeneity stemming from small interstitial spaces and high amounts of biosubstrate.

The P. scaber powder, which was very rich in proteins, was sufficient in

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

The authors extend their appreciation to the Researchers Supporting Project number (RSP-2019/17), King Saud University, Riyadh, Saudi Arabia. We are very grateful to the staff of the J-F. Champollion Center (Albi Campus, France), who provided laboratory facilities during the biomarker analyses.

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