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Growth and Responses of Biomarkers
in the Snail Helix aspersa (Mollusca,
Gastropoda) Used as Bioindicator of Soil
Pollution in Northeast of Algeria
Yousra Bairi, Karima Sifi, and Noureddine Soltani
Keywords
Helix aspersa Sol pollution Biomonitoring Growth Biomarkers
1 Introduction
Among the terrestrial invertebrates to provide information on
soil functions, snails are a good model. Indeed, these species
used in numerous studies as bioindicators, integrate in a
diverse and complementary way the contaminations which
can affect the soil (Leomanni et al. 2015). Our study aims to
biomonitoring of soil quality in Northeast of Algeria where
previous work has shown that metallic pollution predominates
(Larba and Soltani 2014); using the snail Helix aspersa
(Mollusca, Gastropoda) as a bioindicator species. A study of
the growth and response of three biomarkers of environmental
stress; acetylcholinesterase, glutathione S-transferase and
metallothionein, was conducted in this species. Samples were
collected during the four seasons (winter, spring, summer,
autumn) of 2016 at four sites in Northeast of Algeria and
selected according to their level of exposure to pollution; El
Hadjar, site exposed to industrial pollution; El-Tarf and
Bouteldja, sites subject to urban and agricultural pollutions;
and finally El Kala, a site remote from any source of pollution.
2 Material and Methods
2.1 Presentation of Sites
The site of El Kala (36°53′44″N; 8°26′36″E), is located in
extreme east of Algeria close to the Tunisian border, It is
chosen as a reference site because it is distant from any
pollution source. The sites of El Tarf (36°46′2″N; 8°18′50″
E) and Bouteldja (36°30′10″N; 8°06′17″E), are located near
urban and agricultural activities. El Hadjar (36°48′0″N; 7°
43′60″E), is a site submitted to industrial pollution because
is located near a steel complex.
2.2 Species Sampling and Biometric Analysis
Specimens of H. aspersa were collected during the four sea-
sons (winter, spring, summer, autumn) in 2016, from the
selected sites in the Northeast of Algeria. After sampling, H.
aspersa were transported alive to the laboratory, where the
biometric parameters were measured (height, diameter) as well
as the total weight and then each species was dissected and the
head and hepatopancreas were taken for biomarkers analysis.
2.3 Biomarkers Analysis
The activity of acetylcholinesterase (AChE) was determined
in the head as method described by Ellman et al. (1961). The
activity of glutathione S-transferase (GST) and the rates of
metallothionein (MT) were measured in hepatopancreas
according to Habig et al. (1974) and Viarengo et al. (1997)
respectively. The results of the biomarkers were expressed
relative to an amount of protein quantified according to the
method of Bradford (1976).
2.4 Statistical Analysis
The results were expressed as mean ±standard deviation
(SD) and data were tested for normality and homogeneity.
The variation of each parameter among sites and between
seasons was tested by a two-way analysis of variance
Y. Bairi K. Sifi(&)N. Soltani
Laboratory of Applied Animal Biology, Department of Biology,
Faculty of Sciences, University Badji Mokhtar of Annaba, 23000
Annaba, Algeria
e-mail: karima.sifi@univ-annaba.dz; sifi_k23@yahoo.fr
©Springer International Publishing AG 2018
A. Kallel et al. (eds.), Recent Advances in Environmental Science from the Euro-Mediterranean and Surrounding Regions,
Advances in Science, Technology & Innovation, https://doi.org/10.1007/978-3-319-70548-4_107
339
Table 1 Seasonal responses of acetylcholinestease (mM.min
−1
.mg
−1
protein), glutathione S-transferase (mM.min
−1
.mg
−1
protein) and metallothionein (µg MT.mg
−1
protein) in H. aspersa
sampled in four sites in the Northeast of Algeria: El Kala, El Tarf, Bouteldja, El Hadjar during 2016 (mean ±SD; n = 10)
Season AChE GST MT
Site
El Kala El Tarf Bouteldja El Hadjar EL Kala El Tarf Bouteldja El Hadjar El Kala El Tarf Bouteldja El Hadjar
Winter 1.926 ±0.084
aA
1.361 ±0.082
bA
1.448 ±0.051
cA
0.317 ±0.085
dA
0.113 ±0.021
aA
0.201 ±0.061
bA
0.173 ±0.048
bA
0.499 ±0.055
cA
0.170 ±0.012
aA
0.484 ±0.01
bA
0.424 ±0.014
bA
0.863 ±0.061
cA
Spring 1.592 ±0.089
aB
1.024 ±0.014
bB
1.112 ±0.005
cB
0.122 ±0.009
bB
0.152 ±0.002
aA
0.230 ±0.012
bA
0.222 ±0.004
bB
0.558 ±0.038
cB
0.286 ±0.015
aB
0.547 ±0.014
bA
0.500 ±0.018
bA
1.376 ±0.062
cB
Summer 1.275 ±0.060
aC
0.984 ±0.004
bB
1.028 ±0.008
bC
0.091 ±0.002
cB
0.192 ±0.009
aAB
0.472 ±0.056
bB
0.319 ±0.01
cC
0.854 ±0.072
dC
0.477 ±0.041
aC
0.868 ±0.053
bB
0.755 ±0.011
cB
2.098 ±0.290
dC
Autumn 2.002 ±0.015
Aa
1.051 ±0.059
bB
1.196 ±0.069
cD
0.296 ±0.185
dA
0.135 ±0.008
aA
0.416 ±0.007
bC
0.367 ±0.04
cD
0.805 ±0.015
dD
0.223 ±0.006
aAB
0.512 ±0.01
bA
0.619 ±0.017
cC
0.941 ±0.053
dA
Means followed by same letters in minuscule are not significantly different at p > 0.05 between sites within each season; while means followed by same letters in capital are not significantly different at p > 0.05 between seasons within
each site (Tukey’s post hoc test, p < 0.05)
340 Y. Bairi et al.
(ANOVA), followed by Tukey’s post hoc test. All statistical
analysis was performed using GraphPad.Prism.v6. The sig-
nificant difference was defined at p < 0.05.
3 Results and Discussion
Seasonal variation in the growth of H. aspersa revealed that
individuals in the El Kala site are great in size and have
higher weights than individuals of polluted sites of El Tarf,
Bouteldja and El Hadjar where smaller individuals have
been observed in this site. The results of seasonal responses
of AChE, GST and MT showed a significantly inhibition
(p < 0.001) of AChE and a significantly increase
(p < 0.001) of GST and MT in H. aspersa from polluted
sites of El Hadjar, then El Tarf and Bouteldja as compared
with reference site of El Kala (Table 1). Furthermore, the
two-away ANOVA test showed significant effects
(p < 0.001) of both site and season. Our results are agree
with several studies conducted on H. aspersa whose Hispard
et al. (2008) and Larba and Soltani (2014). Indeed, the sites
of El Tarf and Bouteldja are subject to urban and agricultural
pollution, while the El Hadjar site is located near a steel
complex that brings a very important industrial pollution.
4 Conclusion
The results of the present investigation suggest that
H. aspersa is a suitable organism and a sensitive
non-target species that could be used in biomonitoring of
soil pollution based on biomarkers assays. The difference
recorded in the growth and responses of biomarkers
between the different sites located in the Northeast of
Algeria, is related to level of exposition these sites to
pollution.
References
Bradford M. A rapid and sensitive method for the quantification of
microgram quantities of protein utilizing the principle of
protein-dye binding. Anal Biochem. 1976;72:248–54.
Ellman GL, Courtney KD, Andres V, Featherstone RM. A new and
rapid colorimetric determination of acetylcholinesterase activity.
Biochem Pharmacol Physiol Pharm. 1961;38:84.
Habig WH, Pabst MJ, Jacobi WB. The first enzymatic step in
mercapturic acid formation. J Biol Chem. 1974;249:7130–9.
Hispard F, Schuler D, De Vaufleury A, Scheifler R, Badot PM,
Dallinger R. Metal distribution and Metallothionein induction after
cadmium exposure in the terrestrial snail Helix aspersa (Gas-
tropods, Pulmonata). Environ Toxicol Chem. 2008;27(7):1533–42.
Larba R, Soltani N. Use of the land snail Helix aspersa for monitoring
heavy metal soil contamination in Northeast Algeria. Environ Monit
Assess. 2014;186:4987–95.
Leomanni A, Schettino T, Calisi A, Gorbi S, Mezzelani M, Regoli F,
Lionetto MG. Antioxidant and oxidative stress related responses in
the Mediterranean land snail Cantareus apertus exposed to the
carbamate pesticide Carbaryl. Comp Biochem Physiol Part C.
2015;168:20–7.
Viarengo A, Ponzano E, Dondero F, Fabbri R. A simple spectropho-
tometric method for metallothionein evaluation in marine organ-
isms: an application to Mediterranean and Antarctic molluscs. Mar
Environ Res. 1997;44:69–84.
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