Species assemblages and diets of Collembola in the organic matter accumulated over an old tar deposit

Jean-François Ponge

Description

In: European Journal of Soil Biology, 2005, 41 (1-2), pp.39-44. An oil refinery was abandoned in 1964 at Merkwiller-Pechelbrorm (Alsace, France). We investigated the food diets and species assemblages of collembolan communities living in a thick pasty tar deposit overlaid by an organic soil, by comparison with an adjacent unpolluted plot. At the polluted plot the species richness of the springtail community was low (nine species, versus 27 at the unpolluted. plot) but its total abundance was at the same level in both plots. Most springtails consumed more fungi than bacteria at the polluted plot, contrary to the unpolluted plot. Changes in habitat features and food resources might explain the observed decrease in local biodiversity, rather than direct PAH toxicity.

Sujets

• Bacteria
• Diet
• Biodiversity
• Polycyclic aromatic hydrocarbon
• Species richness
• Ecotoxicity
• Springtail
• Fungi

Informations

• Publié par: ponge
• Publié le: 10 avril 2017
• Nombre de lectures: 23
• Langue: English

Extrait

Species assemblages and diets of Collembola in the organic matter accumulated

over an old tar deposit

Email address:jean[e-mail]

habitat features and food resources might explain the observed decrease in local

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AbstractAn oil refinery was abandoned in 1964 at MerkwillerPechelbronn (Alsace,

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fungi than bacteria at the polluted plot, contrary to the unpolluted plot. Changes in

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total abundance was at the same level in both plots. Most springtails consumed more

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Keywords:Collembolan communities; Gut contents; Polycyclic Aromatic Hydrocarbons

Museum National d’Histoire Naturelle, CNRS UMR 5176, 4 avenue du Petit Château,

communities living in a thick pasty tar deposit overlaid by an organic soil, by

comparison with an adjacent unpolluted plot. At the polluted plot the species richness

France). We investigated the food diets and species assemblages of Collembolan

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* Corresponding author. Fax: +33160465009.

Running title:Collembola over tar

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of the springtail community was low (9 species, versus 27 at the unpolluted plot) but its

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91 800 Brunoy, France

biodiversity, rather than direct PAH toxicity.

Servane Gillet, JeanFrançois Ponge*

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1. Introduction

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or benzo[a]pyrene, are toxic aromatic compounds [2,24] which are nonpolar,

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incorporation of litter to mineral matter through earthworm activity [12]. We used a

humus profiles, we observed that the organic matter which accumulated over the 40

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impact of this type of pollution on soil fauna by a comparison between a plot polluted by

patchy pollution of soil and ponds by PAHs. The aim of our work was to study the

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communitylevel approach to evaluate the impact of PAHs on the saprophagous

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yearold tar deposits was permeated by a large population of mycorrhizal fungi, in

and the flexibility of its alimentary habits [11,15,21]. Population indices and food bolus

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indirect effects, for instance through changes in habitat and food quality, are probable

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compartment of the ecosystem. The collembolan community (Arthropoda: Hexapoda)

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their vertical distribution [1,10]. Laboratory bioassays have demonstrated toxic and

waste deposition, even long after site abandonment. Field studies revealed that the

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presence of PAHs decreases the biodiversity of soil animal communities and changes

temperature industrial processes, which induce pollution by accidental oil spilling and

linear, step, or cluster arrangement. Nowadays, their presence in the soil results mainly

hydrophobic and neutral molecules composed of two or more fused benzene rings in

Polycyclic Aromatic Hydrocarbons (PAHs), such as naphthalene, phenanthrene,

anaesthetic effects of PAHs on soil animals [7,5,8], as well as bioaccumulation [17] and

contrast to the adjacent unpolluted plot which was characterized by the rapid

avoidance [18].

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PAHs and a nearby unpolluted control plot. In a parallel study on the composition of

from industrial activities such as coke production, petroleum refining and other high

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[3,11].

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The direct toxicity of pollutants to soil animals has been widely reported and

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Our study site was a petroleum refinery abandoned in 1964. It still displays a

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was chosen for its great abundance and diversity in a wide variety of soil types [19,20]

andTaraxacum officinale, a shrub layer composed ofFraxinus excelsior,Rubus

pendula andQuercus robur. The unpolluted,control plot was located 15 m south of

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buildings now included in a great variety of naturally established ecosystems

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tar, as ascertained by the examination of a number of adjacent (polluted and

composed ofHedera helix,Geranium robertianum,Carex pilosa,Solidago canadensis

deposits. Two plots (polluted versus unpolluted used as control), were sampled in

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October 2002. They were considered to be representative of the patchy occurrence of

analyses were used to follow changes in function and biodiversity of collembolan

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populations under the influence of PAH pollution.

composed of recent fluvial deposits overlaying 1400 m thick sediments above the

granitic substratum [26].

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The study was conducted at the site of the petroleum refinery of Merkwiller

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2.1. Study site

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sand) on the western edge of the Rhine rift valley. The superficial soil (5 m) is

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abandoned. The site is located on the MerkwillerPechelbronn oil field (bituminous

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2. Materials and methods

intensive activity until 1964, afterwards it was progressively dismantled then totally

Pechelbronn, about 50 km north of Strasbourg (Alsace, France). The refinery had an

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a thick pasty tar patch overlaid by a 35 cm organic layer, a discontinuous field layer

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Nowadays, the site area (20 ha) is characterised by the presence of old

2 unpolluted) soil profiles. Thepolluted plot, approximately 20 m , was characterised by

2 the polluted plot, and its surface was arbitrarily fixed to 20 m , for the sake of

fruticosus andSalix capraea and a tree layer composed ofAcer campestre,Betula

(woodland, grassland, ponds) with zones polluted by hydrocarbons, in particular by tar

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identification. The animals were sorted under a dissecting microscope, then mounted in

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abundance, the the species richness, the Shannon Index and the equitability. We also

identified to species level using a phase contrast microscope at 500x and several keys

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comparison, although it did not differ from the rest of the site. It was characterised by a

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much greater plant biodiversity and a good earthworm mull humus. The field layer was

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composed ofHedera helix,Arum maculatum,Carex pilosa,Fragaria vesca,Geranium

robertianum, Geum urbanum,

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canadensis andTaraxacum officinale, the shrub layer was composed ofAcer

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Rubus fruticosus, and the tree layer was composed ofAcer campestre,Prunus avium

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chlorallactophenol (25 ml lactic acid, 50 g chloral hydrate, 25 ml phenol). They were

calculated the total species richness of the five samples taken at each plot.

deposit and in the adjacent unpolluted plot. At each plot, five soil cores, 5 cm diameter

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monogyna,Fraxinus excelsior,Ligustrum vulgare,Prunus avium,Rosa canina and

2.3. Observation of gut contents

Collembola were sampled on the organic matter accumulated over the tar

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and diagnoses [4,6,13,16,23,27,28,29]. We calculated in each sample the total

Potentilla reptans,Stachys sylvatica,Solidago

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by the dryfunnel method [9] and preserved in 95% (v/v) ethyl alcohol until sorting and

and 10 cm depth (litter included), were collected with a core sampler and, after

andQuercus robur.

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discarding tar pieces, stored in plastic bags. Springtails were extracted from soil/litter

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2.2. Sampling and identification of animals

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pseudoplatanus,Carpinus betulus,Cornus mas,Cornus sanguinea,Crataegus

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Samples were collected in April 2003 to determine the amount of PAHs from the

Gut contents from each mounted Collembola were observed under phase

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Holorganic humus (amorphous organic matter)

Mineral particles

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5% in each animal gut then averaged by species and by site.

were taken from each plot, then kept in glass jars and rapidly transported to the

laboratory. Then, soil samples were homogenised and sieved at 1 cm then kept in

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validity of our control plot, samples were collected in May 2003 in the park of the

at 1020 cm depth and (3) the OF horizon overlying the tar deposit. To check the

Hyaline hyphae

Pollen grains

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earthworm mull at neutral pH) under deciduous woody vegetation. Composite samples

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Fungal spores

Melanized hyphae

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laboratory (Brunoy, IledeFrance, France), in the 10 top cm of a similar soil (rich

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The proportion in volume of the different categories was visually estimated to the next

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contrast at 500x magnification and classified in twelve categories:

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2.4. Chemical analyses

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Microalgae

Bacteria

Empty guts

Hemorganic humus (mixture of organic matter and mineral particles)

EPA list [14] in (1) the 10 top cm of the A horizon of the control soil (2) the tar deposit

detector, column LCPAH Supelco). After sieving the contaminated soil, which was

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benzo(ghi)perylene, the others being under detection level (0.02 µg/g for

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acetone (50/50) for soil and acetonitrile for tar. The extract was concentrated under

2.5. Statistical treatment

After extraction of fauna, airdried soil was stored in plastic bags for pH

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Power Liquid Chromatography) with UV detection (alliance 2690 chain, PDA 996

glass jars at 18C° until analysis. Each sample was defrozen, dried, then sieved at 2

mainly made of badly decomposed tree litter, only a small amount of fine matter was

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an estimate of exchange acidity.

PAHs were extracted from each sample with the automatic system ASE 200

measurements were done in triplicate.

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µg/g for contaminated soil, 0.1 µg/g for tar). All

uncontaminated soil, 0.04

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glass electrode. The difference between these two measurements (pH) was taken as

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(Accelerated Solvent Extraction) DIONEX, using a mixture of dichloromethane and

naphthalene, phenanthrene, fluoranthrene, pyrene, benzo(a)anthracene, chrysene and

available for analysis, and only 7 major PAHs could be analysed on this material:

forced air with a TuroVap LV (Zymark), then PAHs were separated by HPLC (High

v:v) for pH H 2 0 and pH KCl, respectively. Each suspension was shaken for five

measurement. The soil was suspended in deionized H 2 0 and 1M KCl (1:5 soil:water

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minutes, then pH was measured in the supernatant after sedimentation for 2 h with a

Nonparametric MannWhitney tests were performed to compare chemical

parameters (pH) and population parameters (abundance, species richness, diversity

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3. Results

from the park of the laboratory (Table 1). As a consequence we estimated that the plot

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chosen as a control at MerkwillerPechelbronn was valid. The total amount of the

were quite similar in the control soil from the MerkwillerPechelbronn site and in the soil

The pH measured in water and potassium chloride was neutral and did not

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the total abundance of springtails did not differ between plots.

control, while the amount of fluoranthrene was only three times higher.

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equitability were higher at the control than at the contaminated plot (Table 2). However,

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3.2. Collembolan communities

differ between both sites at MerkwillerPechelbronn (Table 1). However, mannWhitney

µg/g). The concentration of benzo(ghi)perylene was eleven times higher than the

and equitability of collembolan communities) between sites, using samples as

seven PAHs analysed in the soil over the tar deposit at MerkwillerPechelbronn (5.95

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site.

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polluted zone. The species richness per sample, the Shannon Index and the

The distribution of the 16 PAHs of the EPA list and the total amount of PAHs

At the unpolluted plot 27 species were found whereas only 9 were found in the

µg/g) was six times higher than the corresponding amount at the control plot (0.93

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U test revealed that exchange acidity, expressed bypH, was higher on the unpolluted

3.1. Chemical analyses

replicates (five in each site).

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samples taken at the control plot. The abundance of this species (together with that of

guts was higher at the polluted plot forP. notabilisandS. aureus.

bacteriophagous speciesminimus M.  contained 68% bacteria, and this species was

the low number of specimens (3) collected over the tar deposit. The gut of the

mycelium in the polluted plot. However the latter observation could be an artifact due to

algae, pollen, humus, bacteria and fungal spores and mycelia, but only fungal

was present in all samples taken at the polluted plot while it was absent from all

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candida,Parisotoma notabilis,Sminthurides malmgreni,Sminthurides parvulus and

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only present at the unpolluted site, where it was found in all samples. The isotomidP.

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Sminthurinus aureus. Among these species,S. aureuswas significantly more abundant

represented byProisotoma minutaand (43%) Parisotoma notabilis (40%).P. minuta

Sminthurinus aureus was compared between plots (Figure 1). Fungal mycelium was

inF. candida guts which showed bacteria in equal proportion in both sites but more

more common at the polluted plot, while the contrary was observed for bacteria, except

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Paristoma notabilisin most samples from the tar deposit) explains why the total

3.3. Diets of Collembola

Only five species were common to both polluted and unpolluted plots:Folsomia

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abundance of Collembola did not differ between plots in spite of a much lower number

hyaline hyphae and hemorganic humus over the tar deposit. The proportion of empty

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in the control soil (x 4). Over the tar deposit, 83% of the total abundance was

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The food diet of the three speciesParisotoma notabilis,Folsomia candida and

of species at the polluted plot (Table 2).

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The diet ofS. aureuswas more diverse at the unpolluted plot, containing micro

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use of food resources, since fungi were favoured to the detriment of bacteria. This

influence of pollution by hydrocarbons. This shift can be explained by a change in the

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metals [11], the species richness of springtail communities was observed to decrease

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cases, this could be explained by an increase in free habitats liberated by most

control soil (Table 2). It should be noticed that sieving of the soil to 2 mm before PAH

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minuta, which was present only at the polluted site where it dominated the community,

showed a high proportion of empty guts (67%), 21% mycelium and few bacteria (2%).

speciesProisotoma minutaandParisotoma notabilis.

In our study, changes were observed in the diet of springtails under the

control plot. Three years after application, toxicity was still present given the absence of

Sendstad [25] who found 2% of total Collembola in a polluted plot compared to a

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could be affected by i) the presence of pollutants in the soil, through direct effects of

concentration in the soil over tar. The toxicity of crude oil for Collembola was shown by

observed by Blakely et al. [3]. Here this group was represented by the two dominant

PAHs or other toxic compounds on the most sensitive species and/or ii) indirect effects

sensitive species. Isotomids seemed to be less sensitive to PAH pollution, as was also

under the influence of pollution, at constant or even higher total abundance. In both

4. Discussion

observed between the organic soil accumulated over the tar deposit and the adjacent

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such as changes in food resources. Like in a previous study on a site polluted by heavy

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extraction (see Materials and methods) could have led to underestimate PAH

the tar deposit (Table 1), a great dissimilarity in collembolan communities was

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recolonisation by fauna of the polluted zone. In our study, the collembolan community

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In spite of a slight distance between plots and a low concentration of PAHs over

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possibility reflected a change in the composition of microbial communities in the favour

ectomycorrhizal root mat caused significant changes in the environment of animal as

humus form (and associated foodwebs) than by the direct toxicity of PAHs to

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by

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present at the unpolluted plot. Micromorphological analyses showed a high amount of

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activity)

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of fungi, which could explain why the bacteriophagousMegalothorax minimuswas only

to

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with a poor earthworm activity, while a rapid incorporation of organic matter to mineral

ascertained

well as microbial communities. Given the shift from mull (dominated by earthworm

change in humus form [22].

We found strong changes in invertebrate communities and alimentary habits of

We thank the Agence de l’Environnement et de la Maîtrise de l’Energie (ADEME) for

activity)

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earthworm populations to colonize pollution spots, and the development of a superficial

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microbial communities in favour of fungi could be better explained by a change in

matter through earthworm activity was registered in the control soil [12]. Shifts in

enchytraeid

soil animals forty years after abandonment of industrial activity. Despite a low PAH

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moder

micromorphological analysis [12], the observed decrease in biodiversity and change in

(dominated

concentration in the soil accumulated over the tar deposit (Table 1), the inability of

microorganisms [3].

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Acknowledgements

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ectomycorrizal fungal hyphae in the organic matter accumulated over the tar deposit

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financial support and Vasilica HAMMADE and Laurence BELKESSAM from the Centre

by

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