Physiological and Behavioral Responses of Land Molluscs As Biomarkers For Pollution Impact Assessment - A Review

Journal Pre-proof
Physiological and Behavioral Responses of Land Molluscs as Biomarkers for

Pollution Impact Assessment: A Review
K.S. El-Gendy, A.F. Gad, M.A. Radwan
PII: S0013-9351(20)31455-9
DOI: https://doi.org/10.1016/j.envres.2020.110558
Reference: YENRS 110558
To appear in: Environmental Research
Received Date: 4 September 2020

Revised Date: 4 November 2020
Accepted Date: 28 November 2020
Please cite this article as: El-Gendy, K., Gad, A., Radwan, M., Physiological and Behavioral Responses
of Land Molluscs as Biomarkers for Pollution Impact Assessment: A Review, Environmental Research,
https://doi.org/10.1016/j.envres.2020.110558.
This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition
of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of
record. This version will undergo additional copyediting, typesetting and review before it is published
in its final form, but we are providing this version to give early visibility of the article. Please note that,
during the production process, errors may be discovered which could affect the content, and all legal
disclaimers that apply to the journal pertain.
© 2020 Elsevier Inc. All rights reserved.

Jo
ur
na
lP
re
-p
ro
of
Physiological and Behavioral Responses of Land Molluscs as Biomarkers for Pollution Impact
Assessment: A Review
KS El-Gendy 1, AF Gad 2, MA Radwan 1*
1
Department of Pesticide Chemistry and Technology, Faculty of Agriculture, University of Alexandria,
El-Shatby 21545, Alexandria, Egypt
2
Department of Animal Pests, Plant Protection Research Institute, Agricultural Research Center,
of
Alexandria, Egypt
ro
Abstract
-p
Environmental pollution caused by human activities is a global public health concern. This review
re
discusses the impact of different types of pollutants such as pesticides, metals, nanoparticles and
lP
others on land molluscs. These molluscs are of great interest as good model organisms for studying
biomarker responses in ecotoxicological monitoring programs. Several biomarkers are utilized to

na
characterize and quantify the exposure and harmful impacts of various pollutants. In this review, we
have identified physiological and behavioral endpoints (feeding, growth, avoidance, mucus secretion,
ur
locomotion and reproductive behavior) for the diagnosis of environmental pollution. The present study
Jo
displays that all types of pollutants can disturb physiological and behavioral endpoints of gastropods,
and these impacts depend on the matrix, exposure time and route as well as the type and concentration
of the pollutants in the environment. We have also confirmed that terrestrial gastropods particularly
snails as sentinel species could be used as potential bioindicator organisms for environmental quality
assessment and thus for predicting potential hazards to human health.
Keywords: Pollutants; Biomarkers; Land molluscs; Physiology; Behavior; Bioindicators
1. Introduction
Contamination of the environment by various types of pollutants is a global issue. Due to
urbanization and industrial development, the concentration of these pollutants in the environment
increases at an alarming rate and, therefore, adversely affects human health, non-target organisms and
1
ecosystem sustainability. Recently, with increasing awareness of pollution problems, the improvement
tools of environmental pollution diagnosis have been encouraged. Biological monitoring can be used as
an alternative method for the assessment of pollution and implement short- and long- term strategies
for the protection of environmental resources. It usually depends on the usage of living organisms,
known as “bioindicators” or their responses known as “biomarkers” to indicate environmental changes
(Li et al., 2010).
Terrestrial gastropods, particularly snails are one of the many biological tools used by
scientists as bioindictors for characterising soil quality, determining or predicting damage to
ecosystems and monitoring the pollution (de Vaufleury et al., 2006; Parmar et al., 2016). Such snails
of
possess several criteria; their capability to accumulate various xenobiotics in their tissues, their
ro
sufficient numbers, wide distribution, easy sampling, their tolerant to metals and organic contaminants
-p
and play a significant part in terrestrial food webs (Dallinger et al., 2004; Regoli et al., 2006; Radwan
et al., 2010; de Vaufleury, 2015; Baroudi et al., 2020; Cofone et al. 2020). For these reasons, these
re
animals are increasingly employed as effective sentinel organisms in ecotoxicological monitoring
lP
investigation to determine the impact of various pollutants on the quality of the soil ecosystem based
na
on biomarker responses.
Gastropods are exposed to various contaminants in the environment. It has been shown that
ur
knowledge of the exposure route in terrestrial gastropods is crucial to understanding the toxicity of a
Jo
pollutant (de Vaufleury et al., 2006). These animals that live in close contact with the upper layers of
the soil can be exposed to a truly significant amount of xenobiotics as their foot is in contact with soil
particles and everything else on the surface (Gimbert et al., 2006). In addition, the animals are feed soil
particles when consuming food (Coeurdassier et al., 2002a) or exposed to xenobiotics by inhalation of
air (Regoli et al. 2006).
The measurement of biomarkers has received special attention from researchers and
international agencies as potential new tools for evaluating the harmful effects of contaminants on land
invertebrates (Kammenga et al., 2000; Marigómez et al., 2013). Biomarkers can be defined as
indicators or signals of measurable changes in biological systems at the molecular, biochemical,
cellular, physiological, pathological, or behavioral levels in response to contaminants (Sogorb et al.
2014).
2
Physiological biomarker responses in invertebrate taxa to pollutants act as a biomonitoring
tools for environmental assessments since they may provide a priority over biochemical biomarkers
(Handy and Depledge, 1999). Behavioral biomarkers have also been developed in invertebrates as
monitoring tools for determining the quality of ecosystems (Dell'Omo, 2002; Amiard-Triquet and
Berthet, 2015). Behavioral responses are fully consistent with physiological responses whence
sensibility and efficacy. At the individual level, environmental pollutants can influence
physiological/behavioral functions, such as movement, feeding behavior, and growth of organisms,
which may occur at concentrations significantly below acutely toxic levels. Therefore, such responses
may change the health of an organism, consequently decrease populations and finally lead to dangerous
of
effects on the ecosystem (Hellou, 2011).
ro
In fact, the ecotoxicological effects of pollutants on terrestrial gastropod species with
-p
emphasis on physiological and behavioral endpoints are poorly represented in the scientific literature.
Therefore, this is the first review that provides a comprehensive study on these endpoints that have
re
been developed on terrestrial gastropods as bioindicators for the diagnosis of environmental pollution
lP
and for predicting potential risks to human health. This review also covers the consolidation of three
na
disciplines; ecotoxicology, physiology and behavior in a harmonious framework and presents in a
simple way for tracing a large amount of information on gastropod biomarkers. We believe this will
ur
contribute to the further studies and practical use of these terrestrial gastropod endpoint responses in
Jo
the pollution assessment.
2. Physiological/behavioral biomarkers
Over the few last years, a number of ecotoxicological studies have documented that the
anthropogenic pollutants including pesticides (Table 1) as the most common ones have adverse effects
on human health, non-target organisms and ecosystem. Exposure to pollution can affect higher levels
of biological organization in land gastropods such as reproduction, growth, life history traits and
thereby reduce the gastropod diversity (de Vaufleury, 2015). In addition, toxic substances can have
serious negative impacts on a variety of gastropods, disrupting animal behavior and physiology
(Cofone et al. 2020).
3
Table 1
List of common, trade and chemical names, chemical formula and structure of several classes of pesticides presented in this study
Chemical name
Common name Trade name (s) Class Chemical formula Chemical structure
(IUPAC)
4-(Dimethylamino)-3-
Carbamate
Aminocarb Metacil methylphenyl N- C11H16N2O2
f
Insecticide
oo
methylcarbamate
pr
Gusathion,
e-
3(dimethoxyphosphinothioyl
Guthion, Organophosphorus
Azinphos- methyl sulfanylmethyl)-1,2,3- C10H12N3O3PS2
Pr
Methyl-Guthion, Insecticide
benzotriazin-4-one
Gusathion-M
al
rn
naphthalen-1-yl
Carbaryl Carbamate
Sevin
u
methylcarbamate
Insecticide
C12H11NO2
Jo
2,2-Dimethyl-2,3-dihydro-1-
Furadan, Carbamate
Carbofuran benzofuran-7-yl C12H15NO3
Curater Insecticide
methylcarbamate
4
1-[3,5-dichloro-4-(3-chloro-5- Benzoylphenylurea
triuoromethyl-2-pyridyloxy) Insect Growth
Chlorfluazuron Atabron C20H9Cl3F5N3O3
phenyl]- 3-(2,6-di- Inhibitor
uorobenzoyl)urea
[Cyano-(3-
Cypercot,
phenoxyphenyl)methyl]3-(2,2-
Cypermethrin Ammo, Pyrethroid
dichloroethenyl)-2,2- C22H19Cl2NO3
Cymbush, Insecticide
f
dimethylcyclopropane-1-
oo
Demon
carboxylate
pr
N-[(4- Benzoylphenylurea
e-
Diflubenzuron Dimilin Chlorophenyl)carbamoyl]-2,6- Insect Growth C14H9ClF2N2O2
Pr
difluorobenzamide Inhibitor
al
rn
O,O-dimethyl S-[2-
Dimethoate, Organophosphorus
Dimethoate (methylamino)-2-oxoethyl] C5H12NO3PS2
Rogor
u
dithiophosphate
Insecticide
Jo
6,7-dihydrodipyrido[1,2-a:2′,1′- Bipyridylium
Diquate Reglone C12H12Br2N2
c]pyrazine-5,8-diium dibromide herbicide
Afidan,
Cyclodan, 6,7,8,9,10,10-Hexachloro- Organochlorine
Endosol, 1,5,5a,6,9,9a-hexahydro- 6,9- cyclodiene
C9H6Cl6O3S
Endosulfan Hildan, methano-2,4,3- Insecticide and
Malix, benzodioxathiepine-3-oxide Acaricide
Thiodan
5
Sumithion,
Accothion, O,O-Dimethyl O-(3-methyl-4- Organophosphorus
Fenitrothion C9H12NO5PS
Agrothion, nitrophenyl) phosphorothioate Insecticide
Folithion
Basta,
Rely, (RS)-2-Amino-4-
Glufosinate
Finale, (hydroxy(methyl)phosphonoyl) Herbicide C5H12NO4P
ammonium
Challenge, butanoic acid
f
Liberty
oo
Roundup,
pr
Glyphosate Bypass,
N-(phosphonomethyl)glycine Herbicide C3H8NO5P
e-
Accord,
Glyphomate
Pr
Ariotox,
al
Limatox, 2,4,6,8-tetramethyl-1,3,5,7-
rn
Metaldehyde Molluscicide C8H16O4
Metaldehyde Bait, tetroxocane
Slug Pellets
u
Jo
3,5-Dimethyl-4- Carbamate
Methiocarb Mesurol,
(methylthio)phenyl Insecticide and C11H15NO2S
Mercaptodimethur
methylcarbamate Molluscicide
Lannate,
(E,Z)-methyl N-
Methavin, Carbamate
Methomyl {[(methylamino)carbonyl]oxy} C5H10N2O2S
Methomex, Insecticide
ethanimidothioate
Nudrin
6
Bladan M,
Folidol-M,
O,O-Dimethyl-O-p- Organophosphorus
Parathion methyl Gearphos, C8H10NO5PS
nitrophenylphosphorothioate Insecticide
Metacide,
Metaphos
Gramoxone,
Paraquat Crisquat, 1,1′-Dimethyl-4,4′-bipyridinium Bipyridylium
C12H14Cl2N2
Spot Killer, dichloride herbicide
Sweep
f
oo
Dowicide,
PCP,
pr
Pentachloral,
Pentachlorophenol 2,3,4,5,6-Pentachlorophenol Fungicide C6HCl5O
Pentacon,
e-
Sinituho,
Pr
Weedone
al
(RS)- 1-(4-Chlorophenyl)- 4,4-
Corail,
Tebuconazole dimethyl-3-(1H, 1,2,4-triazol-1- Fungicide C16H22ClN3O
rn
Elite
ylmethyl)pentan- 3-ol
u
Jo
Actara,
3-[(2-Chloro-1,3-thiazol-5-
Thiamethoxam Centric, Neonicotinoid
yl)methyl]-5-methyl-N-nitro- C8H10ClN5O3S
Evident, Insecticide
1,3,5-oxadiazinan-4-imine
Thiomex
Anthon,
Trichlorfon Bovinos, 2,2,2-trichloro-1- Organophosphorus
C4H8Cl3O4P
Chlorophos, dimethoxyphosphorylethanol Insecticide
Dipterex
7
Physiological/behavioral responses are utilized to provide integrated measurements of an
organism’s state, based on a variety of functional characteristics. They are necessary with chemical
support for risk assessment where they can reflect the environmental quality and are well suited to
assess the effects of xenobiotics at sublethal concentrations in the environment (Nicholson et al., 2003).
These biomarkers are ecologically-related indicators that can be associated with the reproduction and
existence of individuals (Crowe et al., 2004). A diagram describing the different physiological and
behavioral endpoint responses of land molluscs to common pollutants is presented in Fig (1).
Terrestrial gastropods are used as model systems for understanding of how general
behavioural processes (such as feeding, locomotion and reproduction) are regulated by the central
of
nervous system (Chase, 2002; Elliott and Susswein, 2002). The nervous system of these animals
ro
involves the central nervous system and the peripheral nervous system. The central nervous system
-p
comprises of ganglia linked by nerve cells. It contains paired buccal ganglia. The paired buccal ganglia
re
include motor neurons, which produce a rhythmical motor output along with moves the diverse buccal
muscles in the harmonic motions needful to feeding (Chase, 2002). The nervous system also plays a
lP
major role in regulating the sexual maturity and sexual behavior in gastropod animals. But, they
na
appear various parts of reproductive physiology that complicate the neurobiology of their
reproduction (Di Cristo and Koene, 2017). Individual neurons can be easily recognized and their
ur
synaptic connection in a neural circuit (networks) can be assessed by simultaneous electrophysiological

Jo
recording from two or more neurons that organize and modulate gastropod feeding. This modulation is
provided by neurotransmitters and peptide cotransmitters (Elliott and Susswein, 2002).
Acetylcholinesterase is an enzyme utilized to estimate the neurotoxicity resulting from xenobiotics
(Durieux et al., 2011). It is a key enzyme in the nervous system, terminating nerve impulses by
catalyzing the hydrolysis of the neurotransmitter acetylcholine. Its alteration is widely used as a
suitable neurological indicator as a result of exposure to various types of pollutants. Additionally, the
neural control of feeding has been studied intensively in a number of gastropod molluscs. In most
gastropods, feeding consists of a variable sequence of food-finding movements, followed by a series of
rhythmic movements in which food is consumed (Elliott and Susswein, 2002). Consequently, changes
in the rate of food consumption due to the exposure of gastropods to pollutants may lead to growth
retardation.
8
Pollutants
Pesticides, Metals,
Nanoparticles, Others
Transfer
Environmental
Compartments
of
ro
Water Soil Air
-p
re
Uptake
lP
na
ur
Jo
Bioaccumulation
Physiological/
Reproduction Behavioral Feeding
Endpoint
Responses
Locomotion Growth
Mucus secretion Avoidance
9
Fig (1): Diagrammatic representation of the different physiological and behavioral endpoint
responses of land molluscs to common pollutants.
Feeding behavior and growth response are generally more sensible than the mortality rate,
as well, appropriate endpoints for assessing the hazard of pollutants ordinarily presented at low
levels in the ecosystem (Gomot-de Vaufleury and Bispo, 2000; El-Gendy et al., 2011; Mleiki et al.,
2016). Summary of studies on feeding and growth indicators in land mollusca species due to
contaminant exposure is listed in Table 2.
2.1. Feeding behavior
Feeding behavior is one of the primary responses to ecological disturbances (McLoughlin et al.,
of
2000), and consequently, the impacts on the feeding of organisms may be extrapolated to impacts on
ro
populations, and then on ecosystem integrity (Slijkerman et al., 2004).
-p
The feeding habits of the land gastropods in the wild (including some litter and prey from dead
re
animals) lead to accumulate toxic substances in their tissues (Dallinger et al., 2001). In addition, the
position of these animals in food webs may play a potential role in transporting chemicals from plants
lP
or waste to carnivores (Notten et al., 2005).

na
The ingestion of methiocarb or metaldehyde has been shown to cause immediate inhibition of
feeding in the grey field slug, Deroceras reticulatum (Bailey et al., 1989); small striped slug, Arion
ur
hortensis, D. reticulatum and longneck field slug, Deroceras caruanae (Wedgwood and Bailey, 1988).
Jo
A significant reduction in the food consumption of the garden snail, Cantareus aspersus (Syn: Helix
aspersa) exposed to carbaryl, azinphos-methyl, fenitrothion, aminocarb, trichlorfon, methyl parathion
and paraquat was found (Schuytema et al., 1994). According to Gomot-de Vaufleury (2000),
pentachlorophenol displayed a sharp decrease in food intake of exposed C. aspersus. The food intake
of C. aspersus after exposure to dimethoate was also reduced (Coeurdassier et al., 2001). Also, the
consumption of Gramoxone® (paraquat) contaminated leaves by the Nigerian land snail, Limicolaria
aurora snails was decreased (Ebenso et al., 2004). Moreover, L. aurora snails treated with Furadan®
(carbofuran) contaminated bait for 120 h, stopped their feeding (Ebenso et al., 2005). As noted by
Godan (1983), Wedgwood and Bailey (1988) and Bailey (2002), it was found that the neurological
effects of carbamate and organophosphorus compounds leading to changes in gastropods feeding
behavior and growth. At higher doses of thiamethoxam, there was no attraction from C. aspersus to
contaminated food (Ait Hamlet et al., 2012), suggesting that may be involved the avoidance behavior
10
which leading to a reduction of food intake. Furthermore, abamectin or thiamethoxam after 2-week
exposure, diminished the food consumption of the white garden snail, Theba pisana (El-Gendy et al.,
2019). Through the first 7 days of C. aspersus exposure to sublethal concentrations of copper
oxychloride, the food intake of snails was minimized without significant differences in food intake
when compared with control during the six weeks of exposure (Snyman, 2001).
Many scientists have reported a significant decrease in food intake of terrestrial gastropods as
the result of metal pollution. For example, heavy metals disrupted several physiological functions of
the Roman snail, Helix pomatia, such as feeding (Rózsa and Salánki, 1990). In a laboratory study, the
food consumption of C. aspersus declined with an increase of Cd concentration in the diet (Russell et
of
al., 1981; Gomot 1997a). It is suggested by the latter authors that Cd might act as a direct toxicant for
ro
altering food consumption through its inhibitory effect on the nerve centers. Cd concentrations in
Urtica dioica leaves did not influence the feeding rate of the grove snail, Cepaea nemoralis, although
-p
its highest concentration caused a significantly lower rate compared to the lowest one (Notten et al.,
re
2006a). About 38% reduction in C. aspersus consumption of food containing high concentration of Zn
lP
was recorded (Simkiss and Watkins, 1990), suggesting that C. aspersus be capable of detecting high Zn
concentrations in the food and decrease its feeding rate if contamination is too high. Marigomez et al.,
na
(1986) reported that the lower concentrations of Cu, Zn, Hg and Pb, did not impact the feeding
ur
behavior of the black slug, Arion ater, however, the higher concentrations reduced food intake.
Similarly, lower concentrations of Pb or Cu did not impact the Limicolaria flammea feeding
Jo
behavior, while higher concentrations caused a slow feeding rate (Amusan et al., 2002). The rise in
consumption rate of metal contaminated food by land gastropods may be due to the inability of animals
to detect low concentrations of contaminants in the food, despite eating. Indeed, these animals manifest
a raise tolerance to contaminated diet with metals possibly due to the binding of metal ions by
metallothioneins and/or intracellular compartmentalization and excretion (Dallinger et al., 1993).
These animals can also regulate metals in their bodies (Berger and Dallinger, 1989). During the 4-
month experiment, Laskowski and Hopkin (1996 a,b) recorded a significant decrease in C. aspersus
consumption of foods contaminated with high concentrations of Cd, Pb, Zn, and Cu. Besides,
Gomot-de Vaufleury (2000) found a sharp decrease in food intake of C. aspersus after Cu, Zn, and Pb
exposure. Swaileh and Ezzughayyar (2000) found that Cu or Cd contaminated diet reduced feeding rate
of the field snail, Helix engaddensis, and unlike Cu, the inhibition produced by Cd was found to be
11
irreversible. Dietary exposure of H. engaddensis to Pb and Zn for one month exhibited significant
reductions in feeding rate (Swaileh and Ezzughayyar, 2001). In addition, the feeding rate of T. pisana
decreases with exposure to Pb, Zn and Cu depending on the dose and exposure time (El-Gendy et al.
2011).
From the aforementioned data, land gastropods can decrease food intake when the metals (Zn,
Cu and Pb) are present in high concentrations in the diet or reach high levels in the body. This
indicates that animals have the ability to detect high concentrations of metals in the food. Reduce of
food intake was also reported in the leatherleaf slug, Semperula maculata after exposure to
HgCl2 up to 96 h (Kamble and Londhe, 2012). This reduction could be related to the neurotoxic
of
action of mercury and to changes in the structure of cerebral ganglia of the animal. Moreover, under
ro
semi-realistic field conditions, C. nemoralis fed on high metal -contaminated leaves collected from a
-p
highly polluted area had significantly lower rates of leaves consumption than those from the reference
area (Notten et al., 2006b). Reduced food intake was also recorded in the giant Ghana African snail,
re
Achatina achatina juveniles' dietary exposed to high levels of Pb (Ebenso and Ologhobo, 2009a), in
lP
transferred snails to dumpsites of battery plant (Ebenso and Ologhobo, 2009b) and in snails exposed to
vehicular exhausts besides roads with various traffic densities (Ebenso and Ologhobo, 2008).
na
Plastics with different sizes have deleterious impacts on terrestrial environment and received
ur
increasing attention (de Souza Machado et al., 2018). It has been shown that microplastic fibres
exposure for 4 weeks caused adverse impact on the fitness of the giant African snail,
Jo
Lissachatina fulica (Syn: Achatina fulica) via reducing their feeding rate (Song et al., 2019). Ingestion
of nanoplastic-contaminated mung bean leaves by L. fulica snails caused a behavioral disorder, as
indicated by a decrease in feeding and foraging speeds of the animals (Chae and An, 2020), indicating
that the impact of these materials attributed to the properties of particles (shape, size, concentration and
availability), the morphology and feeding behavior of snails. Also, feeding consumption of C. aspersus
was declined after exposing animals to Fe2O3-NPs (Besnaci et al., 2016a). Decreased total food intake
was also observed in L. fulica snails after 14 and 28 days exposure to triclosan at 40 mg kg-1 and above
(Wang et al., 2014), demonstrating that this snail is very sensitive to triclosan in terrestrial
environment. Similarly, reduced food consumption rate in T. pisana treated with acrylamide in the diet
for two weeks was detected (El-Gendy et al., 2019).
12
Globally, climate change is the most serious environmental threat that adversely affects
agricultural productivity and the herbivores such as terrestrial gastropods (Bezemer and Knight, 2001).
This climate change mainly caused by greenhouse gases (GHGs) accumulation in the atmosphere,
which results in increased greenhouse effect. These emissions come directly from use of fossil fuels,
tillage practices, fertilized agricultural soils and livestock manure in large proportion. In general,
physiological responses of the herbivores to climate change factors are poorly investigated. For
example, Nicolai and Ansart (2017) reported some insights into physiological and behavioral responses
of terrestrial gastropods to climatic changes: first, terrestrial gastropods use different strategies to
survive sub-zero temperatures in buffered refuges, such as the litter or the soil. The degree of specific
of
cold tolerance influence not only the potential of local extinction, but also of invasion. Second, the
ro
physiological responses to drought and high temperature decrease periods of activity in some species,
thereby reducing foraging and reproduction time. Third, while some communities of terrestrial
-p
gastropods may have a good resilience to fire, storms and flooding, an increase in the frequency of
re
these extreme events could impoverish the community. Fourth, habitat loss and fragmentation.
lP
Schweizer et al. (2019) reported that some snail species are able to cope sever climatic conditions such
as heat and drought through behavioral and physiological adaptations. Indeed, the distribution of land
na
snails has already shifted in recent years due to climate warming (Beltramino et al. 2015) and many of
ur
them will face elevated extinction risk as their habitats become exposed to substantial climatic changes
(Foden et al. 2008).

Jo
CO2 is the greenhouse gas most commonly produced by human activities and it is responsible
for 81% of man-made global warming (EPA, 2018). Plants grown at elevated atmospheric CO2 levels
(600 mg/L air) are eaten at higher rates by H. pomatia snails, this may be due to a decrease in nitrogen
content (Ledergerber et al. 1998), which causes more total consumption.
2.2. Growth response
The growth assessment is also one of the possible indices of physiological integrity (Conti
2008). In ecotoxicology, this index is considered as an integral measurement of the energetic state of
organisms due to pollution stress (Widdows et al., 1997). Growth rate, however, depending upon a
series of physiological and/or biochemical processes, such as changes in the rate of food consumption,
availability, assimilation and nutrients absorption (Sanders et al., 2018). Exposure of terrestrial
13
gastropods to pesticides can cause diminished growth. For example, Schuytema et al., (1994) found a
decrease in snail total weight and shell diameter in C. aspersus dietary exposed to aminocarb,
azinphos-methyl, carbaryl, fenitrothion, methyl parathion, paraquat and trichlorfon for 14 days.
Alterations in the growth rate of T. pisana juveniles intoxicated with sublethal concentrations of
methomyl, dimilin and their combination (1:1) for a period of five weeks were studied by El‐Wakil and
Attia (1999). They observed significant differences in the growth rate of the snails, where, dimilin
treated animals recorded the highest negative growth rate responses. Following four weeks of dietary
exposure of dimethoate, the growth parameters of C. aspersus were reduced with the increase of
exposure time (Coeurdassier et al., 2001), because of the inhibitory effect of this insecticide on AChE
of
activity. Endosulfan (6.25 and 12.50 a.i g/l), applied twice with a month interval to A. achatina,
ro
reduced the growth of snails, while it had little influence on the shell elongation (Wandan et al., 2010),
this effect may be partly due to the influence of the growth substratum. Indeed, the growth rate of T.
-p
pisana was decreased following 14 days of treatment with thiamethoxam and abamectin in the diet (El-
re
Gendy et al., 2019). This effect may be due to the inhibition of carbonic anhydrase enzyme which is
lP
responsible for the shell formation of snails (Zebral et.al., 2019). However, Cypercot® (cypermethrin)
at sublethal doses for 4 weeks of exposure did not impair the growth of the giant West African snail,
na
Archachatina marginata (Amaeze et al., 2011). Under laboratory conditions, Druart et al. (2011)
ur
exposed newly hatched C. aspersus snails for 24 weeks to soil and/or food contaminated with a
formulated product of Bypass® (glyphosate) or Basta® (glufosinate) at the recommended and ten
Jo
times of field dosage rate. They found that these herbicides do not elicit effects on the growth of the
animals. In laboratory and field trials, Snyman (2001) reported that the growth of C. aspersus was
impaired by copper oxychloride treatment.
Metals toxicity was evaluated in different terrestrial gastropod species under laboratory
conditions; for instance, reduced growth of C. aspersus snails was reported after 4 weeks exposure to
high concentrations of Cd in diet or artificial soil (Russell et al. 1981; Gomot 1997b; Coeurdassier et
al., 2002b; ISO, 2006), while growth stimulation was documented in the same species fed with light
Cd-contaminated diet (Gomot 1997b) or soil (Coeurdassier et al. 2002b). Dose-dependent growth
inhibition of exposed S. maculata slugs to Cd was also reported by Kamble and Londhe (2012). In
addition to Cd, some metals like Pb and Zn have exhibited adverse effects on shell size, body weight or
growth of the exposed gastropods in laboratory experiments (Marigómez et al., 1986; Laskowski and
14
Hopkin, 1996a; Gomot-de Vaufleury and Pihan, 2000; Swaileh and Ezzughayyar, 2000, 2001; de
Vaufleury et al. 2006; El-Gendy et al., 2011). Similarly, growth was also reduced in the green garden
snail, Cantareus apertus after 2 weeks of Pb or Cd exposure and more markedly after 4 weeks of
treatment (Mleiki et al., 2016). It can be supposed that the inhibition of growth in gastropod fed on
metal-contaminated food and/or contaminated soil resulted from reducing feeding rate, rejecting
polluted food or disrupting key physiological processes due to competition with promoting growth
factors such calcium and magnesium (Gomot et al., 1989). Calcium (Ca) plays an essential role in the
growth of the entire snail body by increasing the shell mass and thickness, and has been recorded in C.
aspersus (Gomot et al., 1989) and in L. fulica (Ireland, 1991). Such competitive phenomena have been
of
observed between Ca and Pb (Beeby et al., 2002). Exposure to Cd or Pb can also block Ca channels in
ro
snail neurons (Szucs et al. 1994). Moreover, Cd can inhibit GABA-activated ion currents by increasing
intracellular Ca level in snail neurons (Molnár et al. 2004).

-p
It has also been shown that other trace metal contamination of food or contaminated soil leads
re
to a decrease in growth. For example, exposure to Cu-enriched foods caused a decrease of H. pomatia
lP
growth (Moser and Wieser, 1979). The same effects were recorded for other Cu-exposed gastropods
like A. ater (Marigomez et al., 1986), H. engaddensis (Swaileh et al., 2002), C. aspersus (Gomot-de
na
Vaufleury, 2000), L. flammea (Amusan et al., 2002) or T. pisana (El-Gendy et al., 2011). Also,
ur
exposure to Cr or Ni-supplemented foods gave a diminution in C. aspersus aspersus or C. aspersus
maxima growth (Gomot-de Vaufleury and Bispo, 2000; de Vaufleury et al., 2006). In chronic
Jo
exposure, Gimbert et al., (2016) proved the detrimental effect on growth of C. aspersus after exposure
to high Hg++ concentrations via food or soil. Hg++ has led to a stimulation of juvenile growth when the
snails were exposed to low contaminated soil. This phenomenon was clearly evidenced after 28 days of
exposure (+12 %). The same authors suggested that chronic exposure bioassay is not only to estimate
the toxic effects of contaminated soils on snail juveniles but also to detect consequences for
reproductive potential of adults.
Although most laboratory studies have focused on exposure to individual metals, gastropods
are often exposed to contaminant mixtures in the field (Amiard-Triquet and Amiard, 1998). The impact
of metal mixtures on the growth of gastropods can be simply expressed as additive, synergistic or
antagonistic effect (Rainbow, 2002). For example, dietary exposure of C. apertus to a binary mixture of
Cd and Pb caused a decrease in growth by increasing the dose (Mleiki et al., 2016).
15
In field, semi-field or microcosm study, Marigomez et al., (1998) recorded that A. ater slugs
were smaller in the metal-contaminated areas than those in the reference area, and gastropods gathered
from the mining zone displayed the lowest hepatopencreas and whole body weights. Gomot de
Vaufleury and Pihan (2000) reported that the C. aspersus growth was reduced by soil contamination
with Cd, Cu, Pb and Zn after exposure for 4 weeks. Soil contaminated with Cu, Cd, Pb, Ni and Zn
reduced the growth of C. aspersus (Scheifler et al., 2006). After 12-week exposure, Gimbert et al.,
(2008) demonstrated a growth inhibition in C. aspersus when the animal's exposure to the artificial ISO
soils contaminated with 20 and 100 mg Cd/kg, while no marked reduction was noticed for a natural
field soil contaminated with 20 mg Cd/kg. In this field soil, the presence of calcium and magnesium
of
likely explained these contrasting results.
ro
There was a significant reduction in A. achatina growth with increasing exposure to the level
of Pb pollution for 12 weeks due to traffic fumes along roads or at dumpsites near abandoned battery
-p
factory (Ebenso and Ologhobo, 2008; Ebenso and Ologhobo, 2009b). Eeva et al., (2010) showed that
re
the shell mass in different species of snails over a contamination gradient of a copper smelter was
lP
affected, therefore, the tiny shell size indicates low growth of animals in the heavily contaminated
areas. Growth of juveniles was lower in contaminated areas than the control groups, this may be due to
na
the reduction of the growth hormone produced by endocrine or neuroendocrine organs of the snails
ur
(Flari and Edwards, 2003). Grara et al., (2012) revealed that the high concentrations of metal dust- or
Cd-contaminated food had a decrease in body, hepatopancreas, kidney, whole soft tissues and shell
Jo
weights in C. aspersus, this effect can be attributed to the decrease in food intake. Nica et al., (2015)
demonstrated that excessive Cd exposure markedly impacted the shell growth of C. aspersus, and the
shell integrity, however, no consistent impact on shell allometry was detected. Also, C. aspersus
treated with Pb exhibit decreases in shell mass and lateness in shell lip formation (Beeby et al. 2002).
There was a substantial variation in the weights and sizes of shell in C. nemoralis individuals
among polluted sites (Jordaens et al., 2006). Otherwise, Fritsch et al., (2011) declared that no negative
impacts on the body weights as well as the sizes and weights of shell in C. nemoralis and C. aspersus
collected from both polluted smelter and unpolluted sites, however, snails from polluted smelter
produced heavier shells than unpolluted snails under exposure to the highest level of trace metals,
suggesting an adaptive response. Moreover, Boshoff et al., (2015) recorded no morphological
16
alterations in shell strength and thickness of C. nemoralis when the animals treated with either soil or
U. dioica leaves contaminated with metals for 16 weeks. The less sensitivity of C. nemoralis and C.
aspersus to metal pollution may be explained by their adaptation to pollutants that can change the
animal's metabolism, which gives them better ability to cope with the toxicant or genetically adapted;
this can result from selection pressure by a toxicant in a contaminated environment.
In this context, we should be referring to an important point related to the impact of pollutants
on population and community structure of gastropod molluscs. Heavy metal pollution is one of the
anthropogenic activities that affects the survival of native land snail fauna, reduces population
dynamics and their fitness via sublethal effects, and disrupts the diversity and the structure and function
of
of animal community and ecosystems (Hopkin, 1989; Aravind et al., 2005; Douglas et al., 2013). In
ro
highly polluted sites, for example, changes in population and biodiversity of terrestrial gastropods in
neighboring areas of a copper smelter (Eeva et al., 2010), and in mining and industrial areas
-p
(Belhiouani et al., 2019) were reported. An explanation of the lower densities of snails in contaminated
re
areas may be related to metal pollution that decreased the weight of adult snails in the field, possibly
lP
leading to a reallocation of energy resources, from reproductive to metabolic function. Finally, this has
resulted in lower fecundity and hatching success.

na
Further, the L. fulica growth rate was decreased by dietary nanoplastics intake for 14 days
ur
(Chae and An, 2020). However, there is no significant impact on the L. fulica growth indices (shell
diameter and length) after exposure to microplastic fibres for four weeks (Song et al., 2019). Triclosan
Jo
had significant toxic impacts on the biomass and shell diameter growth of L. fulica after two and four
weeks' exposure (Wang et al., 2014) and acrylamide has a marked impact on the T. pisana growth rate
(El-Gendy et al., 2019). Recently, Cofone et al. (2020) reported that defects in the shell structure of the
chocolate-band snail, Massylaea vermiculata (Syn: Eobania vermiculata) after 1-month exposure to
soil amended with nitrates at 2000 mg/L were occurred. Ledergerber et al. (1998) found that H.
pomatia snails fed on plants grown at elevated atmospheric CO2 levels (600 mg/L air) showed a larger
increase in relative wet weight gain than those that fed on plants from ambient CO2 conditions (350
mg/L air).
17
Jo
ur
na
18
lP
re
-p
ro
of
Table 2
List of investigations on feeding and growth biomarkers in land mollusca species exposed to different pollutants
Pollutant Dose/time Exposure type Species Effect Reference

Feeding
Pesticides
Abamectin 0.045 mg/kg
Thiamethoxam 15.69 mg/kg Diet Theba pisana Decrease El-Gendy et al. (2019)
f
oo
/ 2 weeks
0.025 – 0.2 mg/ml
pr
Thiamethoxam Diet Cantareus aspersus No change Ait Hamlet et al. (2012)
/ 6 weeks
e-
50 – 500 mg/ml
Furadan® Diet Limicolaria aurora Stop feeding Ebenso et al. (2005)
/ 5 days
Pr
0.5 – 2.0 %
Gramoxone® Diet Limicolaria aurora Decrease Ebenso et al. (2004)
/ 5 days
al
250 – 6000 mg/kg
Dimethoate Diet Cantareus aspersus Decrease Coeurdassier et al. (2001)
/ 4 weeks
rn
80 and 240 mg/kg
Copper oxychloride Diet Cantareus aspersus No change Snyman (2001)
/ 6 weeks
u
Jo
50 – 1000 mg/kg Gomot-de Vaufleury
Pentachlorophenol Diet Cantareus aspersus Decrease
/ 4 weeks (2000)
Aminocarb
Azinphos- methyl
Carbaryl
5000 mg/kg
Fenitrothion Diet Cantareus aspersus Decrease Schuytema et al. (1994)
/ 2 weeks
Parathion methyl
Paraquat
Trichlorfon
0.5 – 8 %
Methiocarb Baits Deroceras reticulatum Decrease Bailey et al. (1989)
/ 1 week
Metaldehyde 0.5 – 8 % Maize flour Arion hortensis Decrease Wedgwood and Bailey
19
/ 1 week pellets Deroceras. reticulatum (1988)
Deroceras. caruanae
Metals
107 µg/slug Kamble and Londhe
Mercury Contact Semperula maculata Decrease
/ up to 4 days (2012)
50 – 15000 mg/kg
Copper, Lead, Zinc Diet Theba pisana Decrease El-Gendy et al. (2011)
/ 5 weeks
50 –1000 mg/kg Ebenso and Ologhobo
Lead Diet Achatina achatina Decrease
/ 12 weeks (2009a)
f
Industrial
oo
In situ Ebenso and Ologhobo
Lead contaminated Achatina achatina Decrease
/ 12 weeks (2009b)
pr
areas
e-
Lead Traffic fumes Achatina achatina Decrease
/ 12 weeks (2008)
0 and 2.6 mg/kg Decrease at higher
Pr
Cadmium Diet Cepaea nemoralis Notten et al. (2006a)
/ 38 days concentrations
Semi-realistic field
al
Metal contaminated
conditions Diet Cepaea nemoralis Decrease Notten et al. (2006b)
areas
rn
/ 34 days
1– 6 mg/L Decrease at higher
Copper, Lead
/ 3 weeks u Pellets Limicolaria flammea
concentrations
Amusan et al. (2002)
Jo
20 –12500 mg/kg Swaileh and Ezzughayyar
Lead, Zinc Diet Helix engaddensis Decrease
/ 30 days (2001)
Copper 100 –4000
Zinc 4000 – 10000 Gomot-de Vaufleury
Diet Cantareus aspersus Decrease
Lead 100 – 2000 mg/kg (2000)
/ 4 weeks
Cadmium 50 –800
Swaileh and Ezzughayyar
Copper 4 –2500 mg/kg Diet Helix engaddensis Decrease
(2000)
/4 weeks
50 – 800 mg/kg
Cadmium Diet Cantareus aspersus Decrease (>400 µg/g) Gomot (1997a)
/ 4 weeks
20
Zinc 40 –12000
Copper 9 –1600
Decrease at higher Laskowski and Hopkin
Lead 0.4 –12700 Diet Cantareus aspersus
concentrations (1996a,b)
Cadmium 0.16 –145 mg/kg
/ 120 days
1.5 µmoles/g Simkiss and Watkins
Zinc Diet Cantareus aspersus Decrease
/ 4 weeks (1990)
Copper, Lead, 10 – 1000 mg/kg Decrease at higher
Diet Arion ater Marigomez et al. (1986)
Mercury, Zinc / 27 days concentrations
f
I0 – 1000 mg/kg
oo
Cadmium Diet Cantareus aspersus Decrease Russell et al. (1981)
/ 30 days
Metal Oxide Nanoparticles
pr
1, 2 and 3 mg/g
Fe2O3 NPs Diet Cantareus aspersus Decrease Besnaci et al. (2016a)
e-
/ 6 weeks
Pr
Others
Mung bean leaves
10 and 100 mg/kg
al
Nanoplastic grown in Lissachatina fulica Decrease Chae and An (2020)
/ 2 weeks
contaminated soil
rn
10–710 mg/kg soil
Microplastic Contact Lissachatina fulica Decrease Song et al. (2019)
/ 4 weeks
u
Jo
2.28 mg/kg
Acrylamide Diet Theba pisana Decrease El-Gendy et al. (2019)
/ 2 weeks
14 – 340 mg/kg soil
Triclosan Contact Lissachatina fulica Decrease Wang et al. (2014)
/ 2 and 4 weeks
600 mg/L air
Atmospheric CO2 Diet Helix pomatia Increase Ledergerber et al. (1998)
/ 15 days
Growth
Pesticides
Abamectin 0.045 mg/kg
Thiamethoxam 15.69 mg/kg Diet Theba pisana Decrease El-Gendy et al. (2019)
/ 2 weeks
Cypercot® 0.19 – 19.0 ml/L Diet Archachatina marginata No change Amaeze et al. (2011)
21
/ 4 weeks
3 and 30 mg/kg
Bypass® Diet
1.07 and 10.7 mg/kg Cantareus aspersus No change Druart et al. (2011)
Basta® Soil
/ 24 weeks
6.25 and 12.50 g/L
Endosulfan Diet Achatina achatina Wandan et al. (2010)
/ 30 days Decrease
250 – 6000 mg/kg
Dimethoate Diet Cantareus aspersus Decrease Coeurdassier et al. (2001)
/ 4 weeks
80 and 240 mg/kg
Copper oxychloride Diet Cantareus aspersus Decrease Snyman (2001)
f
/ 6 weeks
oo
Methomyl 88.4
pr
Dimilin 1000 mg/kg Diet Theba pisana Decrease El‐Wakil and Attia (1999)
their mixture (1:1) / 5 weeks
e-
Aminocarb
Azinphos- methyl
Pr
Carbaryl
5000 mg/kg
Fenitrothion Diet Cantareus aspersus Decrease Schuytema et al. (1994)
al
/ 2 weeks
Parathion methyl
rn
Paraquat
Trichlorfon
Metals u
Jo
Metal contaminated Mining and Changes in diversity
In situ Different species Belhiouani et al. (2019)
areas industrial areas and population
100 – 10000 Diet
Mercury 50 – 15000 mg/kg Contaminated soil Cantareus aspersus Decrease Gimbert et al. (2016)
/ 13 weeks
Lead 0.025 – 2.5
Cadmium 0.005 – 0.1 mg/g
Their mixtures 0.025 mg Pb + 0.01 mg
Diet Cantareus apertus Decrease Mleiki et al. (2016)
Cd/g and 2.5 mg Pb + 0.1
mg Cd/g
/ 1 and 8 weeks
22
Arsenic, Cadmium, In situ
Copper, Lead, Nickel, Microcosm study Polluted areas Cepaea nemoralis No change Boshoff et al. (2015)
Zinc / 16 weeks
25 – 125 mg/L
/ 30 days followed by Contaminated
Cadmium Cantareus aspersus Decrease Nica et al. (2015)
50-250 mg/L soil
/ 30 days
Changes in diversity
Metal contaminated Anthropogenic
In situ Different species and community Douglas et al. (2013)
areas Activities
f
structure
oo
100 – 1500 mg/kg
Industrial metallic dust Diet Cantareus aspersus Decrease Grara et al. (2012)
/ 4 weeks
pr
107 µg/gastropod Kamble and Londhe
Mercury Contact Semperula maculata Decrease
e-
/ up to 4 days (2012)
50 – 15000 mg/kg
Pr
Copper, Lead, Zinc Diet Theba pisana Decrease El-Gendy et al. (2011)
/ 5 weeks
Metal contaminated Cantareus aspersus
In situ Smelter No change Fritsch et al. (2011)
al
areas Cepaea nemoralis
rn
Metal contaminated
In situ Copper smelter Different species Decrease Eeva et al. (2010)
areas
u Industrial
Jo
Lead contaminated Achatina achatina Decrease
/ 12 weeks (2009b)
areas
20 and 100 mg/kg
Cadmium Contact Cantareus aspersus Decrease Gimbert et al. (2008)
/ 12 weeks
Lead Traffic fumes Achatina achatina Decrease
/ 12 weeks (2008)
75 – 626 Diet
Gomot-de Vaufleury
Cadmium 197 – 1000 mg/kg Contaminated soil Cantareus aspersus Decrease
et al. (2006)
/ 4 weeks
Cadmium, Chromium, Variation
In situ Polluted areas Cepaea nemoralis Jordaens et al. (2006)
Lead, Zinc among sites
Cadmium, Copper, In situ Contaminated soil Cantareus aspersus Decrease Scheifler et al. (2006)
23
Lead, Nickel, Zinc Microcosm study
1 – 6 mg/L
Copper, Lead Pellets Limicolaria flammea Decrease Amusan et al. (2002)
/ 3 weeks
500 mg/kg
Lead Diet Cantareus aspersus Decrease Beeby et al. (2002)
/ 14 weeks
100 – 1000 mg/kg
Cadmium Contact Cantareus aspersus Decrease Coeurdassier et al. (2002b)
/ 4 weeks
Copper 4 – 2500
Cadmium 50 – 800
f
Lead 20 – 12500 Diet Helix engaddensis Decrease Swaileh et al. (2002)
oo
Zinc 20 – 12500 mg/kg
/ 5 weeks
pr
20 – 12500 mg/kg Swaileh and Ezzughayyar
Lead, Zinc Diet Helix engaddensis Decrease
e-
/ 30 days (2001)
Copper 100 – 4000
Pr
Zinc 4000 – 10000 Gomot-de Vaufleury
Diet Cantareus aspersus Decrease
Lead 100 – 2000 mg/kg (2000)
al
/ 4 weeks
rn
Phenanthrene 800
Trichlorophenol 80 Decrease
Pentachlorophenol 80
u Cantareus aspersus
(C. aspersus
Jo
asperses
Cadmium 20 aspersus) Gomot-de Vaufleury and
Diet and contact
Chromium 800 Bispo (2000)
Cantareus aspersus
Lead 800 No change
maxima
Zinc 2000 mg/kg (C. aspersus maxima)
/ 4 weeks
In situ Cantareus aspersus
Cadmium, Copper, / 4 weeks aspersus Gomot-de Vaufleury and
Diet and contact Decrease
Lead, Zinc Cantareus aspersus Pihan (2000)
maxima
Cadmium 50 – 800
Swaileh and Ezzughayyar
Copper 4 – 2500 mg/kg Diet Helix engaddensis Decrease
(2000)
/ 4 weeks
24
Cadmium, Copper, Copper mining
In situ Arion ater Decrease Marigomez et al. (1998)
Zinc areas
50 – 800 mg/kg
Cadmium Diet Cantareus aspersus Decrease Gomot (1997b)
/ 4 weeks
Zinc 40 – 12000
Copper 9 – 1600
Laskowski and Hopkin
Lead 0.4 – 12700 Diet Cantareus aspersus Decrease
(1996a)
Cadmium 0.16 – 145 mg/kg
/ 4 weeks
f
Copper, Lead, 10 – 1000 mg/kg Decrease at higher
oo
Diet Arion ater Marigomez et al. (1986)
Mercury, Zinc / 27 days concentrations
I0 – 1000 mg/kg
pr
Cadmium Diet Cantareus aspersus Decrease Russell et al. (1981)
/ 30 days
e-
20 – 1390 mg/kg Moser and Wieser
Copper Diet Helix pomatia Decrease
/ up to 10 days (1979)
Pr
Others
Mung bean leaves
al
10 and 100 mg/kg
Nanoplastic grown in Lissachatina fulica Decrease Chae and An (2020)
/ 2 weeks
rn
contaminated soil
10– 710 mg/kg soil
Microplastic
/ 4 weeks u Contact Lissachatina fulica No change Song et al. (2019)
Jo
2000 mg/L Changes in shell
Nitrates Contact Massylaea vermiculata Cofone et al. (2020)
/ 30 days structure
2.28 mg/kg
Acrylamide Diet Theba pisana Decrease El-Gendy et al. (2019)
/ 2 weeks
14–340 mg/kg soil
Triclosan Contact Lissachatina fulica Decrease Wang et al. (2014)
/ 2 and 4 weeks
600 mg/L air
Atmospheric CO2 Diet Helix pomatia Increase Ledergerber et al. (1998)
/ 15 days
25
Regarding the determination of other molluscs physiological and behavioral parameters after
exposure to pollutants, four ecologically relevant biomarkers such as avoidance, mucus secretion,
locomotion and reproduction are summarized in Table 3.
2.3. Avoidance behavior
The responses of gastropods to pollutants and the differences demonstrated between the
exposure sources may be primarily explained by a behavioral alteration of organisms in the existence
of the substance. Avoidance as a biomarker is very substantial for behavioral ecotoxicological research
(Loureiro et al., 2009), as it may be observed at sub-lethal or lethal concentrations. This behavioral
of
response is a main parameter associated with different requirements of organisms, including predation,
feeding, migration and reproduction, therefore, tracing and assessing this response allows good
ro
translation of pollutants impact on a population level followed by ecosystem consequences (Loureiro et
-p
al., 2009). Previous studies have documented the avoidance in land gastropods (Schüder et al., 2004;
re
Laskowski and Hopkin, 1996a; Gimbert et al., 2016). This avoidance can be explained as an indirect
toxicity of contaminants, which may be linked to direct toxicity related to the accumulation of toxicant
lP
in gastropod tissues. It has long been known that the copper foil is sometimes used to limit mollusks'
na
access to trees, glasshouse planting benches and pots (Schüder et al., 2003). Copper hydroxide as a
fungicide was found to be feeding deterrent against two slugs; the Florida leatherleaf slug, Leidyula
ur
floridana and the march slug, Deroceras laeve, as well as two snails; the perforate dome snail,
Jo
Ventridens demissus and L. fulica in choice and no-choice laboratory tests (Capinera and Dickens,
2016). Moreover, the avoidance of C. aspersus fed on the hyperaccumulator Senecio coronatus leaves
with nickel was existed (Boyd et al., 2002). At higher levels of augmented CO2 (> 10 mg/L air) paired
with food, T. pisana snails avoided food and climbed away from the food (Lefcort and Kotler, 2017).
2.4. Mucus secretion
The gastropod mucus is a mucous liquid that coats the whole external surface of the animal and
secretes externally by definite salivary epidermal glands located at the snail’s foot (Carriker, 1981).
The mucus has several functions for the animal life, having feeding, locomotion, adherent,
moisturizing, lubricating and protection (South, 1992; Newar and Ghatak, 2015).
HelixComplex snail mucus is characterized by the presence of glycoproteins, sulfated and unsulfated
glycosaminoglycan, allantoin, glycolic acid, polyphenols, sugars, and collagen (Trapella et al. 2018)
26
and exhibited different properties, including antibacterial, antiviral, antifungal and antioxidant activity
(Bortolotti et al. 2016). Two different fractions, water-soluble fraction and the mucin fraction, of L.
fulica snail mucus showed positive antibacterial activity against Gram positive bacteria, such as
Bacillus subtilis and Staphylococcus aureus, and Gram negative strains, like Escherichia coli and
Pseudomonas aeruginosa. Furthermore, the mucin fraction resulted more effective against bacteria
compared to the water-soluble fraction, in which contains Achasin and mytimacin-AF proteins (Iguchi
et al. 1982). Also, the importance of mucus production is to form a protective barrier to prevent
immediate contact between the toxicant and the skin or digestive tract, thereby decreasing the toxicity
of the pollutants (Triebskorn and Ebert, 1989; Triebskorn et al., 1996). The sexual system of
of
gastropods also secretes mucus internally from specific glands (Denny, 1983). Molluscicidal chemicals
ro
e.g. methiocarb and metaldehyde caused mucus stimuli (Godan, 1983; Triebskorn and Ebert, 1989;
Triebskorn et al., 1998). Copper foil, copper-impregnated mats, and copper ammonium carbonate have
-p
been reported to be irritant to terrestrial molluscs thus producing large amounts of mucus and
re
withdrawing their tentacles upon contact with these materials (Schüder et al., 2003; 2004). Likewise,
lP
excessive mucus secretion, especially during 24 h of L. aurora treatment with Gramoxone or Furadan
was recorded (Ebenso, et al., 2004; 2005). On the other hand, maximum mucus secretion was found
na
after 3 and 4 days exposure of the slug S. maculata to HgCl2 and animals did not respond to external
ur
stimuli and exhibited blocked protective action (Londhe et al., 2015).

Jo
2.5. Locomotor behavior
Locomotor behavior is particularly vital to land gastropods life as it facilitates feeding. This
locomotion towards food may also be similar to locomotion away from noxious stimuli. The
locomotion depends upon a combination of muscular actions and the mucus sheet secreted by
the anterior part of the foot (South, 1992; Denny, 1983). The adhesive movement of land gastropods is
a type of animal movement that is realized through a series of muscle contraction waves extending
along a substrate adjacent to the foot surface. The functional unit, that is the foot wave, consists of
muscle and nerve cells. With the versatility of movement and the structure of the pedal ganglia in the
gastropod, it is possible that the production of a variety of well-coordinated wave patterns with the
same sets of muscles requires coordination by a relatively decentralized system of ganglia with several
cross- connections. The presence of a pedal nerve plexus also suggests a considerable degree of
27
peripheral rather than centralized control of pedal waves. In addition, analysis of the mucus present on
the foot during movement showed that the viscosity of mucus during the foot wave is lower than
between waves, which significantly contributes to the effectiveness of this movement (Iwamoto et al.
2014). The velocity of gastropods movement is closely related to the foot wave frequencies, which was
documented in D. reticulatum (Lai et al. 2010) and L. fulica (Kaczorowski et al. 2017).
The locomotor behavior is related to physiological function and can be very sensitive to
chemical exposure. For instance, Rorke et al. (1974) noted that fenitrothion, fenitrothion oxon and
physostigmine at high concentrations reduced locomotion of C. aspersus. Godan (1983) showed that
methiocarb reduced the muscle tone and exerted a toxic effect on the nerve that control locomotion due
of
the accumulation of acetylcholine in synaptic junctions of land molluscs. In the same manner, harmful
ro
effects of metaldehyde on the locomotion of A. hortensis, D. reticulatum and D. caruanae slugs were
-p
observed by Wedgwood and Bailey (1988). Abnormal movement of L. aurora due to Gramoxone®
poisoning has been reported (Ebenso et al., 2004). Also, exposure of L. aurora to Furadan®
re
contaminated pawpaw fruit bait for 120 h ceased snails crawling (Ebenso et al., 2005). On the other
lP
hand, no locomotion was noticed in C. aspersus exposed to the higher concentrations of thiamethoxam,
na
but at lower concentrations, most animals revealed normal activity (Ait Hamlet et al., 2012). Copper
hydroxide was shown to have an effect on the movement of L. floridana towards food (Capinera and
ur
Dickens, 2016). After 24 h of treatment with HgCl2, Londhe et al., (2015) found that the tentacle
Jo
movement of the slug, S. maculata was slowed down, while after 48 h its movements were
comparatively weakened. After 72 h, considerably weaker movement or immobility of the foot and
tentacles was observed, and advertising behavior was completely absent. The alteration of locomotion
behavior in gastropods intoxicated with xenobiotic may be due to the excessive mucus secretion which
risks to dehydration, loss of large amount of energy that needed to cope the toxin stress and/or
neurotoxic effect of a toxicant. Moreover, reduction in locomotor activity has occurred in the small
European land snail, Oxyloma pfeifferi and the brown field slug, Deroceras panormitanum when these
animals were exposed to copper foil (Schüder et al. 2003) or copper impregnated mats and copper
ammonium carbonate (Schüder et al. 2004). In a laboratory experiment with higher levels of
augmented CO2 (> 10 mg/L air) paired with food rewards, Lefcort and Kotler (2017) showed that T.
pisana snails did not change their climbing behavior on lettuce plants when presented with CO2 alone.
28
2.6. Reproductive behavior
Most gastropod molluscs are hermaphrodites and their reproductive system can produce male
and female gametes in the same gonad (ovotestis) or separately in the ovary and the testis (Hodgson,
2009). The presence of mollusc populations relies on the capability of adult molluscs to yield a high
number of offspring (fecundity), therefore, such alteration in fecundity or fertility of the adults due to
the impact of contaminants could severely endanger the individuals, either directly on the free gametes,
or indirectly via accumulation in the reproductive organs (Czech et al., 2001). Developmental and
reproductive toxicity may occur during different stages of life cycle. It is well known that exposure to
toxic substances caused a delay in sexual maturity and subsequently impacted the fecundity of the
of
snail, which proved to be the most sensitive endpoint to contaminants (Lagadic et al. 2007).
ro
Gastropods are ectothermic organisms whose physiological functions like oviposition are
-p
influenced by parasitism, competition, and other environmental conditions (Odiete, 1999; Chase,
re
2002). There are other factors, for example, altitude, food availability, number of essential minerals,
lP
and nutrients that can affect the clutch size and egg viability (Baur and Baur, 1998; Odiete, 1999).
Calcium‐rich soil is a key element in the reproduction of land snails which is transferred from
na
adults to eggs in eggshells and is then utilized to form the shells of the juvenile snails. Egg production
ur
includes the mobilization of Ca from the shell and soft tissue (Tompa, 1980). For instance, slight egg
Jo
production by snails from a polluted site may be a mechanism to keep elevated Ca levels in their tissue,
this may probably minimize their oviposition (Beeby and Richmond, 1988; 2001). Ca-containing
granules are principally shared in the regulation and mobilization of Ca for physiological processes,
particularly shell formation and reproduction (Beeby and Richmond, 1988; Dallinger and Rainbow,
1993).
Ideally, an accurate evaluation of the ecotoxicity of chemicals or contaminated soils should be
based on long-term exposure through full life-cycle tests. Hatching success is an important parameter
in population maintenance. Many publications deal with this important endpoint for aquatic molluscan
species, for example the great pond snail, Lymnaea stagnalis (Gomot 1998; Bandow and Weltje, 2012),
while much less data are available on the embryotoxicity of chemicals or contaminated soils on
terrestrial invertebrates such as snails.
29
Steroid hormones synthesized by gastropod molluscs are very important and play essential
roles in maintaining their reproduction (Kohler et al., 2007; Fernandes et al., 2010). This hormonal
regulation may be disrupted by exposure to various toxic agents, like some natural substances,
pesticides, and industrial chemicals present in the environment. Several studies have been conducted to
evaluate the impacts of these compounds on reproductive performance of terrestrial invertebrates (Hall
et al., 2009). Exposure to these chemicals at ripeness can interfere with the normal cascade of events
leading to eggs and sperm production or affect the normal activity of the genitals at the reproductive
phase. Hormonal unbalances or biochemical interactions caused by xenobiotics may also lead to
reproductive dysfunction in subsequent offspring (Tate et al., 1997).
of
The effect of various pesticides on the reproduction of diverse stages of gastropods like the
ro
number of eggs and egg masses, embryo development as well as hatching has been so far studied.
Druart et al., (2010) investigated the reproduction responses as hatching success and observations of
-p
fetal malformations of C. aspersus after treatment with Reglone (diquat) and Roundup (glyphosate).
re
They noticed fetal abnormalities in unhatched embryos and these effects were toxic and concentration-
lP
dependent. However, Bypass® and Basta® do not interfere with the sexual maturation of C. asperses,
except it reduced the mass of the albumen gland by 43.5% (Druart et al. 2011). Accordingly, the effect
na
on the albumen gland is likely not due to a reduction in food consumption or to metabolic upset, but
ur
rather to an endocrine or neuroendocrine disruption (Gomot-de Vaufleury, 2001). In a liquid phase of
contaminated soil bioassay, Druart et al. (2012) presented that Cabrio® (metiram-zinc +
Jo
pyraclostrobin), Valiant® (cymoxanil + folpet + fosetyl-Al), Corail® (tebuconazole), Roundup® or
Bypass® and Basta® reduced success of egg hatching at concentrations less than that used in the field.
The insect growth regulator, chlorfluazuron induced malformations on the organs of the reproductive
system of M. vermiculata, but the eggs were absent (Mobarak, 2016). The exposure of A. marginata
snails to sublethal concentrations of Cypercot® (cypermethrin) for 28 days, did not induce visible
lesion in the gonadal epithelia (Amaeze et al., 2011). Besides, no significant changes in laying egg
numbers and hatching success after the exposure of C. aspersus snails to copper oxychloride over six-
weeks were observed (Snyman, 2001). On the other hand, high Cu concentrations present in copper
oxychloride gave C. aspersus fecundity dysfunction (Snyman et al., 2009). It is suggested that the
adverse effect on the reproductive success of the snails may be attributed to the accumulation of this
substance in their bodies which imply a major energy cost to cope the stress.
30
According to Rózsa and Salánki (1990), the heavy metal toxicity to H. pomatia leads to the
disturbance of behavior and physiological functions such as reproduction. Higher metal concentrations
in terrestrial gastropods can depressing reproduction (Hopkin, 1989). For instance, Cd exposure can
decline the fertility of C. aspersus snails by minimizing the reproductive vitality (Russell et al., 1981),
delaying the egg-laying cycle and decreasing the number of laid eggs (Gomot-de Vaufleury and
Kerhoas, 2000; Beeby and Richmond, 2001; Gimbert et al., 2008), and inhibiting the hatching of
embryos (Druart et al., 2010, Baurand et al., 2014). Cd may act by interfering with neural and
neuroendocrine factors that regulate genital tract secretion function mainly the albumen gland, as a
marker of endocrine disturbances. Also, Cd can already cross the barrier of the egg and may well reach
of
the embryo and exert its toxicity. Laskowski and Hopkin (1996 a, b) found that the fecundity reduction
ro
in C. aspersus associated with the increase level of Cd, Pb, Cu, and Zn in the diet. Under lab
conditions, the impact of Cu on D. reticulatum or C. aspersus eggs was studied by Iglesias et al. (2000)
-p
and Druart et al. (2012). They deduced that the eggs were greatly sensible to Cu metal or to Cu in
re
Bordeaux mixture, respectively. The effect of metals on fertility may be the result of a disruption of
lP
neuroendocrine processes involved in vitellogenesis and/or egg laying control (Gomot-de Vaufleury,
2001).
na
Lead is facilely lost from the tissues of C. asperses snails, which were handpicked from a
ur
parking lot with a high degree of Pb contamination for up to 102 days, and it did not affect the
oviposition of the populations (Beeby and Eaves, 1983). This might suggest that Pb enable to be
Jo
excreted more rapidly, perhaps as part of the digestive gland faeces. Mercury as an endocrine disruptor
metal (Aljerf and AlMasri, 2018), may lead to fertility dysfunction, impact the role of the
hypothalamic-pituitary-thyroid axis or the hypothalamic-pituitary-adrenal axis, and disrupt steroid
hormones biosynthesis (Georgescu et al., 2011). For example, the reproduction (sexual maturation and
fecundity) of C. aspersus was impaired after Hg++ exposure in food or soil (Gimbert et al., 2016). In
this regard, the higher perturbation occurred in the snails exposed to food compared to soil could be
related to higher Hg++ bioaccessibility and bioavailability to snails via the digestive route. Under field
conditions, no adverse effect on C. nemoralis clutch size was shown from the reference and little
metal-contaminated sites, while animals from the extremely contaminated site did not lay eggs (Notten
et al., 2006b).
31
Further, the deformation of egg membrane of C. aspersus exposed to different concentrations
of Fe2O3 NPs were observed (Besnaci et al. 2016 b). The embryotoxicity of NPs on organisms can be
caused by the in vivo generation of reactive oxygen species (ROS), that result in oxidative stress, which
can be critical for the induction of the observed developmental toxicity (Ozmen et al., 2020).
Alterations in egg hatching and hypopigmentation of M. vermiculata newborn after 1-month exposure
to soil amended with nitrates at 2000 mg/L were reported (Cofone et al. 2020). Additionally, C.
aspersus showed no changes in their life-span when raised at elevated CO2 levels (600 mg/L air) for up
to 38 weeks (Bezemer and Knight, 2001).
Data emerging from this review illustrated that all the above ecologically relevant parameters
of
(feeding behavior, growth, avoidance, movement, mucus production, and reproduction) of land
ro
gastropods are essential and warning signal tools to diagnose the adverse contaminant effects. Any
-p
changes occurred in these endpoints, are likely due to their association with AChE depression, changes
in energy reserves and/or endocrine disruption. In addition, these organisms have a range of biological
re
responses to toxic substances, making them valuable aids in pollution diagnosis and thus was
lP
recommended as useful bioindicators in ecotoxicological research and monitoring programs.

na
ur
Jo
32
Jo
ur
na
33
lP
re
-p
ro
of
Table 3
Summary of studies on avoidance, mucus secretion, locomotion and reproductive indicators in land mollusca species after pollutant exposure
Pollutant Dose/time Exposure type Species Effect Reference

Avoidance
Pesticides
Leidyula floridana
Deroceras laeve Capinera and Dickens,
f
Copper hydroxide 0.96 g /200 ml water Diet Occur
oo
Ventridens demissus (2016)
Lissachatina fulica
pr
Metals
e-
100 – 10000 Diet
Mercury 50 – 15000 mg/kg Contaminated soil Cantareus aspersus Occur Gimbert et al. (2016)
Pr
/ 13 weeks
Copper-impregnated
al
mats, Non-choice /choice tests Deroceras panormitanum
Contact Occur Schüder et al. (2004)
Copper ammonium / 14 hrs Oxyloma pfeifferi
rn
carbonate
Copper foil 7 day u Contact

Deroceras panormitanum
Occur Schüder et al., (2003)
Jo
Oxyloma pfeifferi
Hyperaccumulated
Nickle 37 days Cantareus aspersus Occur Boyd et al. (2002)
leaves
Zinc 40 – 12000
Copper 9 – 1600
Lead 0.4 – 12700 Diet Cantareus aspersus Occur
(1996a)
/ 4 weeks
Others
Atmospheric CO2 10 mg/L air Not occur
˃ 10 mg/L air Diet Theba pisana Occur Lefcort and Kotler (2017)
/ 50 days
34
Mucus secretion
Pesticides
50 – 500 mg/ml
Furadan® Diet Limicolaria aurora Increase Ebenso et al. (2005)
/ 5 days
0.5 – 2.0 %
Gramoxone® Diet Limicolaria aurora Increase Ebenso et al. (2004)
/ 5 days
23 and 63 µg/gastropod
Oral and dermal
Metaldehyde (Oral) / up to 10 hrs Deroceras reticulatum Increase Triebskorn et al. (1998)
application
0.4 % (Dermal) / 1.5 hrs
f
oo
Methiocarb® Pellets 2 % (Oral) / 1 and 5
hrs and 1 % (Contact)
pr
/ 3 hrs Oral and contact Triebskorn and Ebert
Deroceras reticulatum Increase
Metaldehyde® Pellets 2 % (Oral) / 1 and 5 (1989)
e-
hrs and 0.02 % (Contact)
Pr
/ 3 hrs
Metals
al
107 µg/gastropod
Mercury Contact Semperula maculata Increase Londhe et al. (2015)
/ up to days
rn
Copper-impregnated
mats, Non-choice /choice tests
u Contact
Increase Schüder et al. (2004)
Jo
carbonate
Copper foil 7 days Contact Increase Schüder et al., (2003)
Oxyloma pfeifferi
Locomotion
Pesticides
Leidyula floridana
Deroceras leave Capinera and Dickens,
Copper hydroxide 0.96 g / 200 ml water Diet Decrease
Ventridens demissus (2016)
Lissachatina fulica
0.025 – 0.2 mg/ml No locomotion at Ait Hamlet et al.
Thiamethoxam Diet Cantareus aspersus
/ 6 weeks higher concentrations (2012)
35
50 – 500 mg/ml
Furadan® Diet Limicolaria aurora No locomotion Ebenso et al. (2005)
/ 5 days
0.5 – 2.0 %
Gramoxone® Diet Limicolaria aurora Abnormal movement Ebenso et al. (2004)
/ 5 days
Arion hortensis
0.5 – 8 % Wedgwood and Bailey
Metaldehyde Maize flour pellets Deroceras reticulatum Decrease
/ 1 week (1988)
Deroceras caruanae
Fenitrothion
Decrease at higher
Fenitrothion oxon 10 – 250 µg/g gastropod Contact Cantareus aspersus Rorke et al. (1974)
concentrations
f
Physostigmine
oo
Metals
pr
107 µg/gastropod
Mercury Contact Semperula maculata No movement Londhe et al. (2015)
/ up to 4 days
e-
Copper-impregnated
mats, Non-choice /choice tests Deroceras panormitanum
Pr
Contact Decrease Schüder et al. (2004)
carbonate
al
Copper foil 7 day Contact Decrease Schüder et al., (2003)
rn
Oxyloma pfeifferi
Others
u
Jo
Atmospheric CO2 10 mg/L air
No changes in
˃ 10 mg/L air Diet Theba pisana Lefcort and Kotler (2017)
climbing behavior
/ 50 days
Reproduction
Pesticides
1992.5 µg/g Diet Malformation of
Chlorfluazuron 1528.0 µg/ml Contact Massylaea vermiculata reproductive organs Mobarak (2016)
/ 1 week with no eggs
Bordeaux Mixture® 38.2 Liquid, solid or
Reduce the egg
Thiovit® 38.2 gaseous phase of
Cantareus aspersus hatching embryos Druart et al. (2012)
Cabrio® 12.7 contaminated soil
Valiant® 21.2 bioassay
36
Corail® 0.7
Roundup® 5.1
Bypass® 6.1
Basta® 2.1 g/L
/ 20 days
0.19-19.0 ml/L No visible lesion in
Cypercot® Diet Archachatina marginata Amaeze et al. (2011)
/ 4 weeks the gonadal epithelia
No change in sexual
3 and 30 mg/kg
Bypass® Diet maturation
1.07 and 10.7 mg/kg Cantareus aspersus Druart et al. (2011)
f
Basta® Soil Reduce the mass of
oo
/ 24 weeks
the albumen gland
Glyphosate (a,i) 225 – 3600
pr
Roundup® 1.8 – 225
e-
Reglone® 0.375 – 6
Agra 12.5 – 200 Fetal abnormalities in
Pr
Contaminated soil Cantareus aspersus Druart et al. (2010)
Mixture (mg/l of 0.4 – 3 non-hatched embryos
diquat)
al
Cadmium 2- 10 mg/L
/ 1 week
rn
In situ Sprayed vineyard Few oocytes per
Copper oxychloride Cantareus aspersus Snyman et al. (2009)
/ 7 and 60 days
u field 1 mm2 of ovotestis
Jo
80 and 240 mg/kg No change in egg
Copper oxychloride Diet Cantareus aspersus Snyman (2001)
/ 6 weeks numbers and hatching
Metals
100-10000 Diet
Reproduction
Mercury 50-15000 mg/kg Contaminated soil Cantareus aspersus Gimbert et al. (2016)
impaired
/ 13 weeks
Liquid phase of Reduced egg
6 mg /L
Cadmium contaminated soil Cantareus aspersus hatching embryos Baurand et al. (2014)
/ 17 days
bioassay
Decrease in the
20 and 100 mg/kg
Cadmium Contact Cantareus aspersus clutch number and Gimbert et al. (2008)
/ 12 weeks
delay in the egg-
37
laying cycle
Semi-realistic field
Metal contaminated
conditions Diet Cepaea nemoralis No lay eggs Notten et al. (2006b)
areas
/ 34 days
Lead 0 and 500 mg/kg Diet No change
/ 14 weeks Contaminated soil Beeby and Richmond
Cantareus aspersus
Sites with lead In situ No change (2001)
exposure histories
200 and 400 mg/kg Decrease in the Gomot-de Vaufleury and
Cadmium Diet Cantareus aspersus
f
/ 4 weeks number of laid eggs Kerhoas (2000)
oo
Aluminum, Copper, 0.0001 – 1 mg/cm2
Contact Deroceras reticulatum Toxic to eggs Iglesias et al. (2000)
Iron, Zinc / 18-23 days
pr
Zinc 40 – 12000
e-
Copper 9 – 1600
Lead 0.4 – 12700 Diet Cantareus aspersus Reduce fecundity
Pr
(1996a,b)
/ 4 weeks
al
Metal contaminated
> 102 days Car park Cantareus aspersus No oviposition Beeby and Eaves (1983)
rn
areas
I0 – 1000 mg/kg Decrease in
Cadmium
/ 30 days
u Diet Cantareus aspersus
reproductive vitality
Russell et al. (1981)
Jo
Metal Oxide Nanoparticles
Deformation of egg
1.25- 2 mg/L
Fe2O3 NPs Contact Cantareus aspersus membrane at higher Besnaci et al. (2016b)
/ up to 2 weeks
concentrations
Others
2000 mg/L
Nitrates Contact Massylaea vermiculata Egg hatching changes Cofone et al. (2020)
/ 30 days
600 mg/L air No changes in life- Bezemer and Knight
Atmospheric CO2 Diet Cantareus aspersus
/ 12, 25 and 38 weeks span (2001)
38
3. Conclusions and future prospects
As a result of the release of pollutants, which pose a severe threat to human health, non-target
organisms and the environment, there is a great urgency to identify pollutants in the environment and
appraise their impact on populations and communities. The endpoints studies in molluscan species are
important tools for the diagnosis of environmental pollution. Globally, considerable research efforts
have been made to develop parameters in the land molluscs as bioindicators to assess the
environmental pollution.
In this comprehensive review, we have provided researches on the impacts of various types of
contaminants (e.g. pesticides, metals, nanoparticles, micro & nanoplastics and others) on land
of
gastropod species as bioindicators for monitoring changes in soil compartment and their biomarkers as
ro
diagnostic tools for assessing such disturbances in an ecosystem. The present study displays that all
types of pollutants can disturb physiological and behavioral endpoints of gastropods, and these impacts
-p
depend on the matrix, exposure time and route as well as the type and concentration of the pollutants in
re
the environment.
lP
From this review, it is obvious that most studies on the gastropod biomarkers were performed
in laboratory experiments, which elucidate the mechanisms participated in the reaction of gastropod to
na
pollutants. However, their biomarkers have rarely been studied under field conditions that evaluate the
ur
status of gastropod populations in natural conditions. Therefore, field validation of laboratory
exposures should be conducted. Taking into account the fact that extensive background (noise and
Jo
control data) and all natural variations (biotic and abiotic factors) are required to interpret the
significance of biomarkers in organisms collected on site.
Moreover, most studies of gastropod biomarkers concern exposure to some heavy metals,
while organic pollutants are rare. Indeed, all studies were carried out to determine the impacts of a
single pollutant on various species of gastropods. While, under natural environmental conditions,
animals can be exposed to several pollutants at the same time, affecting the physiology of the exposed
individuals. Therefore, ecotoxicity of a mixture of pollutants on gastropod biomarkers should be
performed along with additional studies necessary for the chronic exposure of pollutants to gastropods.
More investigations are also needed to address the impact of endocrine-disturbing compounds
that alter reproductive steroid levels in terrestrial gastropods. Genomic and transcriptomic approaches
as new generation biomarkers can open new windows for understanding some key steps of
39
reproduction in these animals. Furthermore, data on gastropod biomarker responses integrated with
analysis of contaminants in animals and soil will participate in the universal assessment of the pollutant
effects (Radwan et al., 2010, Morin-Crini et al., 2020).
Finally, the comprehensive information derived from this review can provide important
information about environmental quality. It has also been shown that these land molluscs particularly
snails are very useful and efficient for ecotoxicological studies. Future efforts should be made to
integrate various endpoint responses across all organization levels, to investigate and explain endpoint
responses, and to develop acceptable quality procedures. Moreover, study designs should be more
rigorous, including multiple biomarkers that allow a more realistic assessment of pollutants exposure,
of
which in turn can predict, understand and resolve environmental pollution problems worldwide.
ro
References
-p
Ait Hamlet, S., Bensoltane, S., Djekoun, M., Yassi, F., Berrebbah, H., 2012. Histological changes and
re
biochemical parameters in the hepatopancreas of terrestrial gastropod Helix aspersa as
lP
biomarkers of neonicotinoid insecticide exposure. Afr. J. Biotechnol. 11, 16277–16283.

na
Aljerf, L., AlMasri, N., 2018. Mercury toxicity: ecological features of organic phase of mercury in
biota- Part I. Archives of Organic and Inorganic Chemical Sciences 3, 1-8.

ur
Amaeze, N.H., Ugwoeje, D., Egonmwan, R.I., 2011. Histopathological and physiological effects of
Jo
selected agrochemicals on non-target Archachatina marginata. NISEB Journal. 11, 295-302.
Amiard-Triquet, C., Amiard, J.C., 1998. Influence of ecological factors on accumulation of metal
mixtures. Langston, J.W., Bebianno, M., (Eds.), Metal Metabolism in Aquatic
Environments, Chapman and Hall, London, UK, pp. 351-386.
Amiard-Triquet, C., Berthet, B., 2015. Individual biomarkers. In: Aquatic ecotoxicology: advancing
tools for dealing with emerging risks (eds.) Amiard-Triquet, C., Amiard, J-C., Mouneyrac, C.,
Academic Press, pp. 153-182.
Amusan, A.A.S., Anyaele, O.O. Lasisi, A.A., 2002. Effect of copper and lead on growth, feeding and
mortality of terrestrial gastropod Limicolaria Flammea (Müller, 1774). African J. Biomed.
Res. 5, 47–50.
40
Aravind, N.A., Rajashekhar, K.P., Madhyastha, N.A., 2005. Species diversity, endemism and
distribution of land snails of Western Ghats, India; Records of Western Museum Supplement
68, 31-38.
Bailey, S.E.R., 2002. Molluscicicdal baits for control of terrestrial gastropods. In: Molluscs as Crop
Pests (Barker, G. M. ed.) CAB International, London. pp 33-54.
Bailey, S.E.R., Cordon, S., Hutchinson, S., 1989. Why don’t slugs eat more bait? A behavioural study
of early meal termination produced by methiocarb and metaldehyde baits in Deroceras
reticulatum. In: Slugs and Snails in World Agriculture (ed. Henderson, I. F.). Monograph 41.
of
British Crop Protection Council, Thornton Heath. pp. 385-390.
ro
Bandow, C., Weltje, L., 2012. Development of an embryo toxicity test with the pond snail Lymnaea
-p
stagnalis using the model substance tributyltin and common solvents. Sci Total Environ 435–
re
436, 90–95.
lP
Baur, A., Baur, B., 1998. Altitudinal variation in size and composition of eggs in the land snail Arianta
na
arbustorum. Can. J. Zool. 76, 2067-2074.
Baurand, P.E., Capelli, N., Scheifler, R., De Vaufleury, A., 2014. An assessment of the embryotoxicity
ur
of cadmium in the terrestrial mollusk Cantareus aspersus: from bioaccumulation to impacts at

Jo
different levels of biological organization. Ecotox. Env. Saf. 110, 89-94.
Baroudi, F., Al Alam, J., Fajloun, Z., Millet, M., 2020. Snail as sentinel organism for monitoring the
environmental pollution; a review. Ecological Indicators 113, 106240.
https://doi.org/10.1016/j.ecolind.2020.106240
Beeby, A., Eaves, S.L., 1983. Short-term changes in Ca, Pb, Zn and Cd concentrations of the garden
snail Helix aspersa Müller from a central London car park. Environ. Poll. Series A: Ecol.
Biol. 30, 233-244.
Beeby, A., Richmond, L., 1988. Calcium metabolism in two populations of the snail Helix aspersa on a
high lead diet. Archives of Environmental Contamination and Toxicology 17, 507-511.
41
Beeby, A., Richmond, L., 2001. Intraspecific competition in populations of Helix aspersa with
different histories of exposure to lead. Environ. Pollut. 114, 337-344.
Beeby, A., Richmond, L., Herpé, F., 2002. Lead reduces shell mass in juvenile garden snails (Helix
aspersa). Environ. Pollut. 120, 283-288.
Belhiouani, H., El-Hadef EL-Okki, M., Afri-Mehennaoui, F.Z., Sahli, L., 2019. Terrestrial gastropod
diversity, distribution and abundance in areas with and without anthropogenic disturbances,
Northeast Algeria. Biodiversitas 20, 243-249.
Beltramino, A.A., Vogler, R.E., Gutiérrez Gregoric, D.E., Alejandra Rumi, A., 2015. Impact of climate
of
change on the distribution of a giant land snail from South America: predicting future trends
ro
for setting conservation priorities on native malacofauna. Climatic Change 131, 621–633.
-p
Berger, B., Dallinger, R., 1989. Accumulation of cadmium and copper by terrestrial snail Arianta
re
arbustorum L.: kinetics and budgets. Oecologia 79, 60-65.
lP
Besnaci, S., Bensoltane, S., Braia, F.M.H., Zerari, L., Khadri, S., Loucif, H., 2016 b. Embryotoxicity
evaluation of iron oxide Fe2O3 on land snails: Helix aspersa. J. Entomol. Zool. Stud. 4, 317-
na
323.
ur
Besnaci, S., Bensoltane, S., Zerari, L., Samia, C, Ait Hamlet S., Berrebbah, H., 2016 a. Impact of
Jo
nanometric iron oxide in the hepatopancreas of terrestrial gastropod Helix aspersa:
histological changes and biochemical parameters. International Journal of Pharmaceutical
Sciences Review and Research, 36, 234–241.
Bortolotti, D., Trapella, C., Bernardi, T., Rizzo, R., 2016. Letter to the editor: antimicrobial properties
of mucus from the brown garden snail Helix aspersa. Br J Biomed Sci. 73, 49–50.
Boshoff, M., Jordaens, K., Baguet, S., Bervoets, L., 2015. Trace metal transfer in a soil-plant-snail
microcosm field experiment and biomarker responses in snails. Ecological indicators 48, 636-
648.
42
Boyd, R.S., Davis M.A., Wall, M.A., Balkwill, K., 2002. Nickel defends the South African
hyperaccumulator Senecio coronatus (Asteraceae) against Helix aspersa (Mollusca:
Pulmonidae). Chemoecology. 12, 91–97.
Bezemer, T.M., Knight, K.J., 2001. Unpredictable responses of garden snail (Helix aspersa)
populations to climate change. Acta Oecologica 22, 201–208.
Capinera, J.L., Dickens, K., 2016. Some effects of copper-based fungicides on plant-feeding terrestrial
molluscs: A role for repellents in mollusc management. Crop Protection 83, 76-82.
of
Carriker, M.R., 1981. Shell penetration and feeding by naticacean and muricacean predatory
gastropods: a synthesis. Malacologia 20, 403-422.
ro
-p
Chae, Y., An, Y. J., 2020. Nanoplastic ingestion induces behavioral disorders in terrestrial snails:
trophic transfer effects via vascular plants. Environ. Sci.: Nano 7, 975-983.
re
lP
Chase, R., 2002. Behavior and its Neural Control in Gastropod Molluscs. New York: Oxford
University Press
na
Coeurdassier, M., Gomot-de Vaufleury, A., Lovy, C., Badot, P.M., 2002b. Is the cadmium uptake
ur
from soil important in bioaccumulation and toxic effects for snails? Ecotox. Environ. Saf. 53,
Jo
425-431.
Coeurdassier, M., Gomot-de Vaufleury, A., Saint-Denis, M., Ribera, D., Narbonne, J.F., Badot, P.M.,
2002a. Effects of dimethoate on snail B-esterase and growth as a function of dose, time and
exposure route in a laboratory bioassay. Biomarkers 7,138–150.
Coeurdassier, M., Saint-Denis, M., Gomot-de Vaufleury, A., Ribera, D., Badot, P.M., 2001. The
garden snail (Helix aspersa) as a bioindicator of organophosphorus exposure: Effects of
dimethoate on survival, growth, and acetylcholinesterase activity. Environ. Toxicol. Chem.
20, 1951–1957.
43
Cofone, R., Carraturo, F., Capriello, T., Libralato, G., Siciliano, A., Del Giudice, C., Maio, N., Guida,
M., Ferrandino, I., 2020. Eobania vermiculata as a potential indicator of nitrate contamination
in soil. Ecotoxicology and Environmental Safety 204,111082.
Conti, M.E., 2008. Biological Monitoring: Theory & Applications: Bioindicators and Biomarkers.
Technology & Engineering, p.228.
Crowe, T.P., Smith, E.L., Donkin, P., Barnaby, D.L., Rowland, S.J., 2004. Measurements of sub lethal
effects on individual organisms indicate community-level impacts of pollution. J. Appl. Ecol.
41, 114-123.
of
Czech, P., Weber, K., Dietrich, D. R., 2001. Effect of endocrine modulating substance on reproduction
ro
in the hermaphroditic snail Lymnaea stagnalis L. Aquatic Toxicol. 53, 103-114.
-p
Dallinger, R., 1993 Strategies of metal detoxification in terrestrial invertebrates. In: Dallinger, R.,
re
Rainbow, P.S., (eds.), Ecotoxicology of Metals in Invertebrates. Boca Raton: Lewis
lP
Publishers; 246–289.
na
Dallinger, R., Berger, B., Triebskorn, R., Köhler, H., 2001. Soil biology and ecotoxicology. In: The
Biology of Terrestrial Molluscs, Barker, G.M. (Ed.). CABI Publishing, New York, pp. 489-
ur
525.
Jo
Dallinger, R., Lagg, B., Egg, M., Schipflinger, R., Chabicovsky, M., 2004. Cd accumulation and Cd-
Metallothionein as a biomarker in Cepaea hortensis (Helicidae, Pulmonata) from laboratory
exposure and metal-polluted habitats. Ecotoxicology 13, 757-772.
Dallinger, R., Rainbow, P.S., 1993. Ecotoxicology of metals in invertebrates. Lewis Publishers, Boca
Raton.
Dell’Omo, G., 2002. Behavioural Ecotoxicology. John Wiley & Sons, Chichester, UK.
Denny, M. W., 1983. Molecular biomechanics of molluscan mucous secretions. In Wilbur, K., Simkiss,
K., Hochachka, P. W. (eds.), The Mollusca. Academic Press, New York.
44
de Souza Machado, A.A., Kloas, W., Zarfl, C., Hempel, S., Matthias C Rillig, M.C., 2018.
Microplastics as an emerging threat to terrestrial ecosystems. Global Change Biology
24,1405 –1416.
de Vaufleury, A., 2015. Land snail for ecotoxicological assessment of chemicals and soil
contamination-ecotoxicological assessment of chemicals and contaminated soils using the
terrestrial snail, Helix aspersa, at various stage of its life cycle: a review, Chapter 23, 365-366.
In: Armon, R.H., Hännninen, O. (eds) Environ Indicators. Springer, Berlin.
de Vaufleury, A., Coeurdassier, M., Pandard, P., Scheifler, R., Lovy, C., Crini, N., Badot, P.M., 2006.
of
How terrestrial snails can be used in risk assessment of soils. Environ Toxicol Chem 25,797-
ro
806.
-p
Di Cristo, C., Koene, J.M., 2017. Neurobiology of reproduction in molluscs: mechanisms and
re
evolution. The Oxford Handbook of Invertebrate Neurobiology, Byrne, J.H. (ed.).
DOI:10.1093/oxfordhb/9780190456757.013.26.
lP
Douglas, D. D., Brown, D. R., Pederson, N., 2013. Land snail diversity can reflect degrees of
na
anthropogenic disturbance. Ecosphere 4, 28. http://dx.doi.org/10.1890/ES12-00361.1

ur
Durieux, E.D. H., Farver, T.B., Fitzgerald, P.S., Eder, K.J., Ostrach, D.J., 2011. Natural factors to
Jo
consider when using acetylcholinesterase activity as neurotoxicity biomarker in Young-Of-
Yearstriped bass (Morone saxatilis). Fish Physiol Biochem 37, 21-29.
Druart, C., Millet, M., Scheifler, R., Delhomme, O., de Vaufleury, A., 2011. Glyphosate and
glufosinate-based herbicides: fate in soil, transfer to and effects on land snails. J Soils
Sediment 11,1373–1384.
Druart, C. Scheifler, R., de Vaufleury, A., 2010. Towards the development of an embryotoxicity
bioassay with terrestrial snails: screening approach for cadmium and pesticides. J. Hazard
Mater. 184, 26-33.
45
Druart, C., Scheifler, R., Millet, M., de Vaufleury, A., 2012. Land snail eggs bioassays: A new tool to
assess embryotoxicity of contaminants in the solid, liquid or gaseous phase of soil. Appl. Soil
Ecol. 53, 56-64.
Ebenso, I. E., Chukwu S., Udo S., Ukpai, C. U., Udo, M. T., Umanah, U., Isang, A., Akam, A., 2004.
Some behavioural responses of Limicolaria aurora exposed to gramoxone. Niger. J. Physiol.
Sci. 19, 82-85.
Ebenso, I.E., Ita, B., Umoren, E.P., Ita, M., Binang, W., Edet, G., Izah, M., Udo, I.O., Ibanga, G.,
Ukpong, E.E., 2005. Effect of carbamate molluscicide on african giant land snail Limicolaria
of
aurora. J. Appl. Sci. Environ. Mgt. 9, 99-102.
ro
Ebenso, I.E., Ologhobo, A.D., 2008. Effects of lead pollution from vehicular exhaust fumes against
-p
sentinel juvenile Achatina achatina. Bull. Environ. Contam. Toxicol. 81, 513-515.
re
Ebenso, I.E., Ologhobo, A.D., 2009a. Effects of lead pollution against juvenile Achatina achatina fed
lP
on contaminated artificial diet. Bull. Environ. Contam. Toxicol. 82, 106-110.
Ebenso, I.E., Ologhobo, A.D., 2009b. Effects of lead pollution at industrial contaminated sites on
na
sentinel juvenile Achatina achatina. Bull. Environ. Contam. Toxicol. 82, 583-585.
ur
Eeva, T., Rainio, K., Suominen, O., 2010. Effects of pollution on land snail abundance, size and
Jo
diversity as resources for pied flycatcher, Ficedula hypoleuca. Sci. Total Environ. 408, 4165-
4169.
El-Gendy, K.S., Radwan, M.A., Gad, A.F., 2011. Feeding and growth responses of the snail Theba
pisana to dietary metal exposure. Arch Environ Contam Toxicol. 60, 272-280.
El-Gendy, K.S., Radwan, M.A., Gad, A.F., Khamis, A.E., Eshra, E.H., 2019. Physiological traits of
land snails Theba pisana as simple endpoints to assess the exposure to some pollutants.
Environ. Sci. Pollut. Res. 26, 6922-6930.
Elliott, C.J.H., Susswein, A. J., 2002. Comparative neuroethology of feeding control in molluscs.
Journal of Experimental Biology 205, 877-896.
46
El‐Wakil, H.B., Attia, A.M., 1999. Effect of selected insecticides on terrestrial snails Eobania
vermiculata (Müller) and Theba pisana (Müller) with respect to some morphological changes
in Egypt. J. Environ. Sci. Health B. 34, 47-60.
EPA (Environmental Protection Agency), 2018. Overview of Greenhouse Gases, Carbon Dioxide
Emissions.
Fernandes, D., Loi, B., Porte, C., 2010. Biosynthesis and metabolism of steroids in molluscs. J.
Steroid Biochem. Mol. Biol. 127, 189-195.
Flari, V.A., Edwards, J.P., 2003. The role of the endocrine system in the regulation of reproduction in
of
terrestrial pulmonate gastropods. Invertebr. Reprod. Dev. 44, 139-161.
ro
Foden, W., Mace, G., Vié, J.C., Angulo, A., Butchart, S., DeVantier, L., Dublin, H., Gutsche, A.,
-p
Stuart, S., Turak, E., 2008. Species susceptibility to climate change impacts. In: Vié, J.C.,
re
Hilton-Taylor, C., Stuart, S.N. (eds) The 2008 review of the IUCN red list of threatened
lP
species. IUCN Gland, Switzerland, pp 1–11
Fritsch, C., Coeurdassier, M., Gimbert, F., Crini, N., Scheifler, R., de Vaufleury, A., 2011.
na
Investigations of responses to metal pollution in land snail populations (Cantareus aspersus

ur
and Cepaea nemoralis) from a smelter-impacted area. Ecotoxicology 20, 739-759.

Jo
Georgescu, B., Georgescu, C., Dărăban, S., Bouaru, A., Paşcalău, S., 2011. Heavy metals acting as
endocrine disrupters. Anim Sci Biotechnol. 44, 89-93.
Gimbert, F., de Vaufleury, A., Douay, F., Coeurdassier, M., Scheifler, R., Badot, P.M., 2008. Long-
term responses of snails exposed to cadmium-contaminated soils in a partial life-cycle
experiment. Ecotoxicol. Environ. Saf. 70, 138-146.
Gimbert, F., de Vaufleury, A., Douay, F., Scheifler, R., Coeurdassier, M., Badot, P.M., 2006.
Modelling chronic exposure to contaminated soil. A toxicokinetic approach with terrestrial
snail Helix aspersa. Environ Int 32, 866–875.
Gimbert, F., Fanny, P., Caire, A.L., de Vaufleury, A., 2016. Mercury toxicity to terrestrial snails in a
partial life cycle experiment. Environ Sci Pollut Res., 23, 3165-3175.
47
Godan, D., 1983. Pest Slugs and Snails: Biology and Control. Springer-Verlag, Berlin/ New York.
Gomot, A., 1997a. Effects of heavy metals on snail development. Use of snails as bio-indicators of
heavy metal pollution for the preservation of human health. Bull Acad. Natl. Med.181, 59-74.
Gomot, A., 1997b. Dose-dependent effects of cadmium on the growth of snails in toxicity bioassays.
Arch. Environ. Contam Toxicol. 33, 209-216.
Gomot, A., 1998. Toxic effects of cadmium on reproduction, development and hatching in the
freshwater snail Lymnaea stagnalis for water quality monitoring. Ecotoxicol Environ Saf 41,
288–297.
of
ro
Gomot, A., Gomot, L., Boukraa, S., Bruckert, S.,1989. Influence of soil on the growth on the land snail
Helix aspersa. An experimental study of absorption route for the stimulating factors. J. Moll.
Stud., 55, 1-7.

-p
re
Gomot-de Vaufleury, A., 2000. Standardized growth toxicity testing (Cu, Zn, Pb, and
lP
Pentachlorophenol) with Helix aspersa. Ecotoxicol. Environ. Saf. 46, 41-50.

na
Gomot-de Vaufleury, A., 2001. Regulation of growth and reproduction. In: Barker, G.M. (ed). The
Biology of Terrestrial Molluscs. CABI, Oxon, pp 331–355.

ur
Jo
Gomot-de Vaufleury, A., Bispo, A., 2000. Methods for toxicity assessment of contaminated soil by oral
or dermal uptake in land snails. 1. Sublethal effects on growth. Environ Sci Technol. 34,
1865-1870.
Gomot de Vaufleury, A., Kerhoas, I., 2000. Effects of cadmium on the reproductive system of the land
snail, Helix aspersa. Bull. Environ. Contam. Toxicol. 64, 434-442.
Gomot de Vaufleury, A., Pihan, F., 2000. Growing snails used as sentinels to evaluate terrestrial
environment contamination by trace elements. Chemosphere. 40, 275-284.
Grara, N., Atailia A., Boucenna M., Berrebbah H., Djebar, M-R., 2012. Toxicity of metal dust from
Annaba steel complex (Eastern Algeria) on the morpho- physiological parameters of the snail
Helix aspersa. Adv. Environ Biol. 6, 605-611.
48
Hall, C. M., Rhind, S.M., Wilson, M.J., 2009. The potential for use of gastropod molluscs as
bioindicators of endocrine disrupting compounds in the terrestrial environment. J. Environ.
Monit.11, 491-497.
Handy, R.D., Depledge, M.H., 1999. Physiological responses: their measurement and use as
environmental biomarkers in ecotoxicology. Ecotoxicology. 8, 329-349.
Hellou, J., 2011. Behavioural ecotoxicology, an “early warning” signal to assess environmental
quality. Environ Sci Pollut Res Int. 18,1-11.
of
Hodgson, AN., 2009. Reproduction and sex in invertebrates. In: da Silva AP (ed.), Reproduction and
Development Biology. Encyclopedia of Life Support System. Developed under the auspices
ro
the UNESCO, EOLSS Publishers, Paris, France, pp 1–8.
-p
Hopkin, S.P., 1989. Ecophysiology of metals in terrestrial invertebrates. Elsevier Applied Science, NY,
re
USA, 366 p.
lP
Iglesias, J., Castillejo, J., Parama, R., Mascato, R., Lombardia, M.J., 2000. Susceptibility of the eggs of
na
the pest slug Deroceras reticulatum to contact with metal salts. J. Molluscan Stud. 66, 171-
176.
ur
Jo
Iguchi, S.M., Aikawa, T., Matsumoto, J.J., 1982. Antibacterial activity of snail mucus mucin. Comp
Biochem Physiol A. 72, 571–574.
Ireland, M.P., 1991. The effect of dietary calcium on growth, shell thickness and tissue calcium
distribution in the snail Achatina fulica. Comp. Biochem. Physiol., 98A,111–116.
ISO, 2006. Soil quality – Effects of pollutants on juvenile land snails (Helicidae) – Determination of
the effects on growth by soil contamination, ISO 15952. International Organization for
Standardization, Geneva, Switzerland.
Iwamoto, M., Ueyama, D., Kobayashi, R., 2014. The advantage of mucus for adhesive locomotion in
gastropods. J. Theoret. Biol. 353, 133-141.
49
Jordaens, K., De Wolf, H., Vandecasteele, B., Blust, R., Backeljau, T., 2006. Associations between
shell strength, shell morphology and heavy metals in the land snail Cepaea nemoralis
(Gastropoda, Helicidae). Science of the Total Environment 363, 285-293.
Kaczorowski, P., Piskorska, E., Olszewska-Slonina, D., Grzeœk, G., 2017. Adhesive movement
characteristics of the snail Achatina fulica. Folia Biologica (Kraków) 65, 133-135.
Kamble, N.A., Londhe, S.R., 2012. Mercuric chloride induced neuropathology and
physiological responses in slug Semperula maculata. Int. J. Zool. Res. 8, 33-42.
Kammenga, J.E., Dallinger, R., Donker, M.H., Köhler, H.R., Simonsen, V., Triebskorn, R., Weeks,
of
J.M., 2000. Biomarkers in terrestrial invertebrates for ecotoxicological soil risk assessment.
ro
Rev. Environ. Contam. Toxicol. 164, 93-147.
-p
Kohler, H. R., Kloas, W., Schirling, M., Lutz, I., Reye, A. L., Langen, J. S., Triebskorn, R., Nagel, R.,
re
Schönfelder, G., 2007. Sex steroid receptor evolution and signalling in aquatic
lP
invertebrates. Ecotoxicology 16, 131-143.

na
Lagadic, L., Coutellec, M-A., Caquet, T., 2007. Endocrine disruption in aquatic pulmonate molluscs:
few evidences, many challenges. Ecotoxicology 16, 45–59.

ur
Lai, J. H., del Alamo, J. C., Rodriguez-Rodriguez, J., Lasheras, J. C., 2010. The mechanics of the
Jo
adhesive locomotion of terrestrial gastropods. Journal of Experimental Biology 213, 3920
DOI: 10.1242/jeb.046706
Laskowski, R., Hopkin, S.P., 1996a. Effect of Zn, Cu, Pb, and Cd on fitness in snails (Helix aspersa).
Ecotoxicol. Environ. Saf. 34, 59-69.
Laskowski, R., Hopkin, S.P., 1996b. Accumulation of Zn, Cu, Pb and Cd in the garden snail (Helix
aspersa): Implications for predators. Environ Pollut. 91, 289-297.
Ledergerber, S., Leadley, P.W., Stocklin, J., 1998. Feeding behaviour of juvenile snails (Helix
pomatia) to four plant species grown at elevated atmospheric CO2. Acta Oecol. 19, 89–95.
50
Lefcort, H., Kotler, B.P., 2017. Life in a near-future atmosphere: carbon dioxide enrichment increases
plant growth and alters the behavior of a terrestrial snail but not a terrestrial beetle. Israel
Journal of Ecology and Evolution 63(3), doi.10.1163/22244662-06301008
Li, L., Zheng, B., Liu, L., 2010. International Conference on Ecological Informatics and Ecosystem
Conservation (ISEIS 2010) Biomonitoring and bioindicators used for river ecosystems:
definitions, approaches and trends. Procedia Environ. Sci. 2,1510-1524. doi:
10.1016/j.proenv.2010.10.164.
Londhe, S., Patil, S., Kamble, N., 2015. Effect of mercuric chloride on terrestrial slug Semperula
of
maculata and histopathology of reproductive organs: a comprehensive Study. Toxicological
ro
and Environmental Chemistry, 97, 170-185.
-p
Loureiro, S., Amorim, M.J.B., Campos, B., Rodrigues, S.M.G., Soares, A.M.V.M., 2009. Assessing
re
joint toxicity of chemicals in Enchytraeus albidus (Enchytraeidae) and Porcellionides
pruinosus (Isopoda) using avoidance behaviour as an endpoint. Environmental Pollution 157,

lP
625-636.
na
Marigomez, I.A., Angulo, E., Saez, V., 1986. Feeding and growth responses to copper, zinc, mercury
and lead in the terrestrial gastropod Arion ater (Linne). J. Moll. Stud. 52, 68-78.
ur
Jo
Marigómez, I. A., Garmendia, L., Soto, M., Orbea, A., Izagirre, U., Cajaraville, M.P., 2013. Marine
ecosystem health status assessment through integrative biomarker indices: a comparative
study after the Prestige oil spill “mussel watch.”. Ecotoxicology. 22, 486-505.
Marigomez, I.A., Kortabitarte, M., Dussart G.B.J., 1998. Tissue level biomarkers in sentinel slugs as
cost-effective tools to assess metal pollution in soils. Arch Environ. Contam. Toxicol. 34, 167-
176.
McLoughlin, N., Yin, D., Maltby, L., Wood, R.M., Yu, H. 2000. Evaluation of sensitivity and
specificity of two crustacean biochemical biomarkers. Environ. Toxicol. Chem. 19, 2085-
2092.
Mleiki, A., Irizar, A., Zaldibar, B., El-Menif, N.T., Marigómez, I., 2016. Bioaccumulation and tissue
distribution of Pb and Cd and growth effects in the green garden snail, Cantareus apertus
51
(Born, 1778), after dietary exposure to the metals alone and in combination. Sci. Total
Environ. 547, 148-156.
Mobarak, S.A., 2016. The malformation effect of caprice (chlorfluazuron) compound on the
reproductive system of land snail Eobania vermiculata. J Basic Appl Zool 74, 51–55.
Molnár, G., Salánki, J., Kiss, T., 2004. Cadmium inhibits GABA-activated ion currents by increasing
intracellular calcium level in snail neurons. Brain Res., 1008, 205-211.
Morin-Crini, N., Luzon, M., Amiot, C., de Vaufleury, A., 2020. QuEChERS applicability to measure
land snail polycyclic aromatic hydrocarbons (PAHs) for risk assessment. Toxicological &
of
Environmental Chemistry 102, 209-223. https://doi.org/10.1080/02772248.2020.1781855.
ro
-p
Moser, H., Wieser, W., 1979. Copper and nutrition in Helix pomatia (L.). Oecologia 42, 241-251.
re
Newar, J., Ghatak, A., 2015. Studies on the adhesive property of snail adhesive mucus. Langmuir 31,
lP
12155–12160.
Nica, D.V., Filimon, M.N., Bordean, D-M, Harmanescu, M., Draghici, G.A., Dragan, S., Gergen, I.I.,
na
2015. Impact of soil cadmium on land snails: A two-stage exposure approach under semi-field
ur
conditions using bioaccumulative and conchological end-points of exposure. PLoS One,

Jo
10(3), e0116397. https://doi.org/10.1371/journal.pone.0116397
Nicholson, B.L., Perakis, A.N., Bulkley, J.W., 2003. Environmental assessment, Sea born
Petrochemical Spill Analysis Within the United States, 1992-1999, Environmental
Management, Vol 31, No 4.
Nicolai, A., Ansart, A., 2017. Conservation at a slow pace: terrestrial gastropods facing fast-changing
climate. Conserv Physiol. 5(1), cox007. doi: 10.1093/conphys/cox007.
Notten, M.J., Oosthoek, A.J., Rozema, J., Aerts R., 2005. Heavy metal concentrations in a soil-plant-
snail food chain along a terrestrial soil pollution gradient. Environ. Pollut. 138, 178-190.
52
Notten, M. J., Oosthoek, A. J., Rozema, J., Aerts, R., 2006a. The land snail Cepaea nemoralis regulates
internal Cd levels when fed on Cd-enriched stinging nettle (Urtica dioica) leaves at low,
field-relevant concentrations. Environ. Pollut. 139, 296-305.
Notten, M.J., Oosthoek, A.J., Rozema, J., Aerts, R., 2006b. Heavy metal pollution affects consumption
and reproduction of the land snail Cepaea nemoralis fed on naturally polluted Urtica dioica
leaves. Ecotoxicol. 15, 295-304.
Odiete, W.O., 1999. Environmental physiology of Animals and pollution. Diversified resources LTD.
Ozmen, M., Gungordu, A., Geckil, H., 2020. Ecotoxicity of nanomaterials in aquatic environment. In:
of
Thangadurai, D., Sangeetha, J., Prasad, R. (eds) Nanotechnology for Food, Agriculture,
ro
and Environment. Nanotechnology in the Life Sciences, pp.351-377. Springer, Cham.
-p
https://doi.org/10.1007/978-3-030-31938-0_15
re
Parmar, T. K., Rawtani, D., Agrawal, Y. K., 2016. Bioindicators: the natural indicator of environmental
lP
pollution. Front Life Sci. 9, 110-118.

na
Radwan, M.A., El-Gendy, K.S., Gad, A.F., 2010. Biomarkers of oxidative stress in the land snail,
Theba pisana for assessing ecotoxicological effects of urban metal pollution. Chemosphere.
ur
79, 40-46.
Jo
Rainbow, P.S., 2002. Trace metal concentrations in aquatic invertebrates: why and so what? Environ.
Pollut., 120, 497-507.
Regoli, F., Gorbi, S., Fattorini, D., Tedesco, S., Notti, A., Machella, N., Bocchetti, R., Benedetti,
M., Piva, F., 2006. Use of the land snail Helix aspersa as sentinel organism for monitoring
ecotoxicologic effects of urban pollution: an integrated approach. Environ. Health Perspect.
114, 63-69.
Rorke, M.A., Gardner, D.R., Greenhalgh, R., 1974. Lethality and behavioural symptoms produced by
some organophosphorous compounds in the snail (Helix aspersa). Bull. Environ. Contam.
Toxicol. 11, 417-424.
53
Russell, L.K., DeHaven, J.I., Botts, R.P., 1981. Toxic effects of cadmium on the Garden snail (Helix
aspersa). Bull. Environ. Contam. Toxicol, 26, 634-640.
Rózsa, K., Salánki, J., 1990. Heavy metals regulate physiological and behavioral events by modulating
ion channels in neuronal membranes of molluscs. Environ. Monitor. Assess. 14, 363-365.
Sanders, T., Schmittmann, L., Nascimento-Schulze, J.C., Melzner, F., 2018. High calcification costs
limit mussel growth at low salinity. Front. Mar. Sci. 5, 352. doi: 10.3389/fmars.2018.00352
Scheifler, R., de Vaufleury, A., Coeurdassier, M., Crini, N., Badot, P.-M., 2006. Transfer of Cd, Cu,
Ni, Pb, and Zn in a soil-plant-invertebrate food chain: A microcosm study. Environ.
of
Toxicol. Chem. 25, 815-822.
ro
Schüder, I., Port, G., Bennison, J., 2003. Barriers, repellents and antifeedants for slug and snail control.
Crop Protection 22, 1033-1038.

-p
re
Schüder, I., Port, G., Bennison, J., 2004. The behavioural response of slugs and snails to novel
lP
molluscicides, irritants and repellents. Pest Manag Sci 60,1171-1177.

na
Schuytema, G.S., Nebeker, A.V., Griffis, W.L., 1994. Effects of dietary exposure to forest pesticides
ur
on the brown garden snail Helix aspersa Müller. Arch. Environ. Contam. Toxicol. 26, 23-28.
Jo
Schweizer, M., Triebskorn, R., Köhler, Heinz‐R., 2019. Snails in the sun: Strategies of terrestrial
gastropods to cope with hot and dry conditions. Ecology and Evolution 9, 12940–12960.
Simkiss, K., Watkins, B., 1990. The influence of gut microorganisms on zinc uptake in Helix aspersa.
Environ. Pollut. 66, 263-271.
Slijkerman, D.M.E., Baird, D.J., Conrad, A., Jak, R.G., van Straalen, N.M., 2004. Assessing structural
and functional plankton responses to carbendazim toxicity. Environ. Toxicol. Chem. 23, 455-
462.
Snyman, R.G., 2001. Copper oxychloride, in the common garden snail Helix aspersa, in Western Cape
Vineyards. PhD Thesis, University of Stellenbosch.
54
Snyman, R.G., Reinecke, S.A., Reinecke, A.J., 2009. Quantitative changes in digestive gland cells and
oocytes of Helix aspersa, as biomarkers of copper oxychloride exposure under field
conditions. Bull Environ Contam Toxicol 83,19-22.
Sogorb, M.A., Estevez, J., Vitanova, E., 2014. Biomarkers in biomonitoring of xenobiotics. Chapter
57, pp. 965-973. In: Biomarkers in Toxicology, Gupta, R.C. (eds) Elsevier, Academic Press.
Song, Y., Cao, C., Qiu, R., Hu, J., Liu, M., Lu, S., He, D., 2019. Uptake and adverse effects of
polyethylene terephthalate microplastics fibers on terrestrial snails (Achatina fulica) after soil
exposure. Environ. Pollut. 250, 447-455.
of
South, A., 1992. Terrestrial slugs: Biology, ecology and control. Chapman & Hall., London, UK, 428
ro
p.
-p
Swaileh, K.M., Ezzughayyar, A., 2000. Effects of dietary Cd and Cu on feeding and growth rates of the
re
land snail Helix engaddensis. Ecotoxicol. Environ. Saf. 47, 253-260.
lP
Swaileh, K.M., Ezzughayyar, A., 2001. Dose-dependent effects of dietary Pb and Zn on feeding and
growth rates of the land snail Helix engaddensis. Ecotoxicol. Environ. Saf. 50, 9-14.
na
Swaileh, K.M., Hussein, R., Halaweh, N., 2002. Metal accumulation from contaminated food and its
ur
effect on growth of juvenile land snails Helix engaddensis. J. Environ. Sci. Health B. 37,
Jo
151-159.
Szucs, A., Salenki, J., S-Rozsa, K., 1994. Effect of chronic exposure to cadmium-or lead-enriched
environments on ionic current of identified neurons in Lymanaea stagnalis L. Cell Mol.
Neurobiol. 14, 769-780.
Tate, T.M., Spurleck, J.O., Christian, F.A., 1997. Effects of glyphosate on the development of
Pseusosuccinea columnella snails. Arch. Environ. Contam. Toxicol. 33, 286–289.
Tompa, A.S., 1980. Studies on the reproductive biology of gastropods: Part III. Calcium provision and
the evolution of terrestrial eggs among gastropods. J. Conchol. 30, 145–154.
Trapella, C., Rizzo, R., Gallo, S., Alogna, A., Bortolotti, D., Casciano, F., Zauli, G., Secchiero, P.,
Voltan, R., 2018. HelixComplex snail mucus exhibits pro-survival, proliferative and pro-
55
migration effects on mammalian fibroblasts. Sci Rep. 8, 17665.doi: 10.1038/s41598-018-
35816-3.
Triebskorn, R., Christensen, K., Heim, I., 1998. Effects of orally and dermally applied metaldehyde on
mucus cells of slugs (Deroceras reticulatum) depending on temperature and duration of
exposure. J. Moll. Stud. 64, 467-487.
Triebskom, R., Ebert, D., 1989. The importance of mucus production in slug's reaction to molluscicides
and the impact of molluscicides on the mucus producing system. In: Henderson, I.F. (ed)
Slugs and Snails in World Agriculture. BCPC Monograph 41, Thornton Heath, pp 373–379.
of
Triebskorn, R., Henderson, I.F., Martin, A., Kohler, H.-R., 1996. Slugs as targets or non-target
ro
organisms for environmental chemicals. In: Slug and Snail Pests in Agriculture (Henderson,
-p
I.F. ed.). British Crop Protection Council Monograph, 66, 65-72.
re
Wandan, E.N., Elleingand, E.F., Koffi, E., Clément, B.N., Charles, B., 2010. Impact of the
lP
insecticide endosulfan on growth of the African giant snail Achatina achatina (L.). Afr. J.
Environ. Sci. Technol. 4, 685-690.

na
Wang, X., Liu, Z., Wang, W., Yan, Z., Zhang, C., Wang, W., Chen, L., 2014. Assessment of toxic
ur
effects of triclosan on the terrestrial snail (Achatina fulica). Chemosphere 108, 225-230.
Jo
Wedgwood, M.A., Bailey, S.E.R., 1988. The inhibitory effects of the molluscicide metaldehyde on
feeding, locomotion and faecal elimination of three pest species of terrestrial slug. Annals
Applied Biol. 112, 439-457.
Widdows, J., Nasci, C., Fossato, V.U., 1997. Effects of pollution on the scope for growth of mussels
(Mytilus galloprovinciallis) from the Venice Lagon, Italy. Mar. Environ. Res. 43, 69-79.
Zebral, Y.D., da Silva Fonseca, J., Marques, J.A., Bianchini, A., 2019. Carbonic anhydrase as a
biomarker of global and local impacts: Insights from calcifying animals. Int J Mol Sci. 20(12),
3092.
56
Jo
ur
na
57
lP
re
-p
ro
of
Declaration of interests
☒ 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.
☐The authors declare the following financial interests/personal relationships which may be considered
as potential competing interests:
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.
of
ro
-p
re
lP
na
ur
Jo

Physiological and Behavioral Responses of Land Molluscs As Biomarkers For Pollution Impact Assessment - A Review

Uploaded by

Copyright:

Available Formats

You might also like

Physiological and Behavioral Responses of Land Molluscs As Biomarkers For Pollution Impact Assessment - A Review

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Physiological and Behavioral Responses of Land Molluscs As Biomarkers For Pollution Impact Assessment - A Review

Uploaded by

Copyright:

Available Formats

Journal Pre-proof

Physiological and Behavioral Responses of Land Molluscs as Biomarkers for

K.S. El-Gendy, A.F. Gad, M.A. Radwan

To appear in: Environmental Research

Received Date: 4 September 2020

© 2020 Elsevier Inc. All rights reserved.

KS El-Gendy 1, AF Gad 2, MA Radwan 1*

biomarker responses in ecotoxicological monitoring programs. Several biomarkers are utilized to

assessment and thus for predicting potential hazards to human health.

Keywords: Pollutants; Biomarkers; Land molluscs; Physiology; Behavior; Bioindicators

Contamination of the environment by various types of pollutants is a global issue. Due to

known as “bioindicators” or their responses known as “biomarkers” to indicate environmental changes

(Li et al., 2010).

scientists as bioindictors for characterising soil quality, determining or predicting damage to

air (Regoli et al. 2006).

indicators or signals of measurable changes in biological systems at the molecular, biochemical,

cellular, physiological, pathological, or behavioral levels in response to contaminants (Sogorb et al.

physiological/behavioral functions, such as movement, feeding behavior, and growth of organisms,

disciplines; ecotoxicology, physiology and behavior in a harmonious framework and presents in a

the pollution assessment.

(Cofone et al. 2020).

synaptic connection in a neural circuit (networks) can be assessed by simultaneous electrophysiological

provided by neurotransmitters and peptide cotransmitters (Elliott and Susswein, 2002).

Acetylcholinesterase is an enzyme utilized to estimate the neurotoxicity resulting from xenobiotics

gastropods, feeding consists of a variable sequence of food-finding movements, followed by a series of

Mucus secretion Avoidance

contaminant exposure is listed in Table 2.

2.1. Feeding behavior

or waste to carnivores (Notten et al., 2005).

aspersa) exposed to carbaryl, azinphos-methyl, fenitrothion, aminocarb, trichlorfon, methyl parathion

effects of carbamate and organophosphorus compounds leading to changes in gastropods feeding

metallothioneins and/or intracellular compartmentalization and excretion (Dallinger et al., 1993).

of nanoplastic-contaminated mung bean leaves by L. fulica snails caused a behavioral disorder, as

for two weeks was detected (El-Gendy et al., 2019).

(Foden et al. 2008).

content (Ledergerber et al. 1998), which causes more total consumption.

2.2. Growth response

impaired by copper oxychloride treatment.

intracellular Ca level in snail neurons (Molnár et al. 2004).

exposure to Cr or Ni-supplemented foods gave a diminution in C. aspersus aspersus or C. aspersus

reproductive potential of adults.

suggesting an adaptive response. Moreover, Boshoff et al., (2015) recorded no morphological

this can result from selection pressure by a toxicant in a contaminated environment.

resulted in lower fecundity and hatching success.

Pollutant Dose/time Exposure type Species Effect Reference

locomotion and reproduction are summarized in Table 3.

2.3. Avoidance behavior

2.4. Mucus secretion

stimuli and exhibited blocked protective action (Londhe et al., 2015).

2.5. Locomotor behavior

Ideally, an accurate evaluation of the ecotoxicity of chemicals or contaminated soils should be

terrestrial invertebrates such as snails.

reproductive dysfunction in subsequent offspring (Tate et al., 1997).

rather to an endocrine or neuroendocrine disruption (Gomot-de Vaufleury, 2001). In a liquid phase of

pyraclostrobin), Valiant® (cymoxanil + folpet + fosetyl-Al), Corail® (tebuconazole), Roundup® or

hypothalamic-pituitary-thyroid axis or the hypothalamic-pituitary-adrenal axis, and disrupt steroid

to 38 weeks (Bezemer and Knight, 2001).