Article

pubs.acs.org/Langmuir

Studies on the Adhesive Property of Snail Adhesive Mucus

Janu Newar and Archana Ghatak*
KIIT School of Biotechnology, KIIT University, Bhubaneswar 751024, India
ABSTRACT: Many gastropod molluscs are known to secrete mucus which allow
these animals to adhere to a substrate while foraging over it. While the mucus is
known to provide strong adhesion to both dry and wet surfaces, including both
horizontal and vertical ones, no systematic study has been carried out to understand
the strength of such adhesion under dierent conditions. We report here results
from preliminary studies on adhesion characteristics of the mucus of a snail found
in eastern India, Macrochlamys indica. When perturbed, the snail was found to
secrete its adhesive mucus, which was collected and subjected to regular adhesion
tests. The hydrated mucus was used as such, and also as mixed with buer of dierent pH. These experiments suggest that the
mucus was slightly alkaline, and showed the maximum adhesion strength of 9 kPa when present in an alkaline buer. Preliminary
studies indicate that adhesive force is related to the ability of the mucus to incorporate water. In alkaline condition, the gel like
mass that it forms, incorporate water from a wet surface and enable strong adhesion.

INTRODUCTION
Adhesives of dierent kinds, such as polysaccharides, proteins,
and the mixture of both are secreted by a vast range of
organisms. A general term describing these polymeric materials
is biological adhesives, most of which provide strong adhesion
to a substrate in hostile environmental conditions.1 An example
is the biological adhesives from molluscs such as mussels, which
have received scientic attention because of their unique ability
to provide underwater attachment, not provided by many manmade adhesives.2 In recent times many studies have also
focused on gastropod molluscs, that is, snails and slugs, since
most of them can attach very strongly on vertical surfaces.
Snails and slugs secrete trail mucus in order to carry out
variety of functions: locomotion and attachment on horizontal
and vertical surfaces, self-defense, locating prey items, and
mating.1,3,4 The work of Smith and his co-workers introduced a
dierent type of mucus called adhesive mucus.1,5 Although
secreted for similar purposes as trail mucus, adhesive mucus can
provide stronger attachment to a substrate than trail mucus.3
Irrespective of their use, adhesive mucus from dierent
gastropods possesses similar characteristics; they all are more
viscous and stier than normal trail mucus; they are all mixtures
of proteins and polysaccharides containing more proteins than
the corresponding trail mucus;5 some of these proteins are
metal binding proteins and demonstrate metal dependent
oxidative properties.6 A number of biochemical studies in terms
of composition, oxidative reactions, viscosity, and hardness57
of this type of mucus have been reported from a few species of
gastropods, however reports on the performance of this mucus
as an adhesive material are lacking. Importantly it is not clear
yet what gives these materials their ability to stick to wet
surfaces very strongly, as well as on a variety of substrates,
rough and smooth alike. Knowledge of such an adhesion
mechanism will be helpful in designing a biocompatible
universal adhesive that works in both dry and wet environments. It is in this context that we present our studies with a
XXXX American Chemical Society

common garden snail from India whose mucus was not

previously known to be an adhesive material. This snail,
Macrochlamys indica, was rst described by Blanford and
Godwin-Austen (1908).8 In recent times, ecological studies on
this snail has been done by Jahan et al. (2002)9 and karyotype
analysis has been done by El-Alfy et al. (1994).10 Our
experiments showed the mucus secreted by this snail is in
fact a strong adhesive which works excellently well on wet
surfaces. By using a standard adhesion test we have examined
and quantied the adhesion strength of this mucus at variety of
conditions which reveal a rather distinct feature of this
adhesive: its ability to absorb water in specic state which
corresponds to its ability to adhere to a wet surface, something
not achieved in man-made adhesives. We have alsdo found out
the biochemical feature of the adhesive which is responsible for
such active control of adhesion.

EXPERIMENTAL SECTION

Materials. Snails were collected from Eastern India, Kolkata, West

Bengal. Flower pots, earthenware bowls, and plants were bought from
local markets. All the buers, SDS-PAGE reagents, and protein gel
silver-staining reagents were bought from HIMEDIA Laboratories,
Mumbai, India. Instrument for measuring adhesive force was made by
local workshop. Microscope glass slides and weight box for common
balance was used for adhesive force measurement.
Maintenance of the Snail in the Laboratory. Snails were kept
in owering pots, of approximately 10 in. of height, containing small
owering plants. About 5 in. of space was left in the pot above the soil
so that snails could climb up the wall of the pot when they wanted.
The pots were covered with a net so that they could not escape. Snails
were found to feed on the leaves of owering plants like Marigold,
hibiscus, and coleus. In addition to plant leaves they were also fed fresh
vegetables such as papaya, bottle gourd, ridged gourd, ash gourd,
pointed gourd, and pumpkin (Figure 1(a)). Small earthenware bowls
were kept in the pots in order to provide the snails places to hide.
Received: March 1, 2015

DOI: 10.1021/acs.langmuir.5b03498
Langmuir XXXX, XXX, XXXXXX

Article

Langmuir

slide in a way that the slides form a cross with an overlapping area of
6.25 sq. cm. Within minutes the combination of slides attached by the
wet mucus is placed on an instrument and measurement of adhesive
force was started. The adhesive force is the force required to pull apart
the glass slides. The instrument used for measurement of attachment
force was made in the local workshop (see Figure 2 for the diagram

Figure 1. (a) Snails maintained in owering pots feeding on

vegetables. (b) The snail releasing yellow adhesive mucus during
eeing.
These snails come out of their hiding place when it is dark and feed
during the night.
Water was sprinkled once or twice a day to keep the soil moist and
the environment humid. Temperature control was not applied, it
varied from 18 to 38 C throughout the year. Hygienic conditions
were maintained in and around the pots to minimize the growth of
fungus along with prevention of ants and mites. Pots were cleared of
dead animals and rotten leaves and vegetables every day. From time to
time, these pots were kept in strong sunlight after removing all the
snails from it. Laying of eggs by the snails was observed twice a year in
NovemberDecember and AprilMayJune. The newly born snails
took almost 3 months to fully grow into adult snails.
Collection of Snail Mucus and Subsequent Studies. Snails
were taken out of the pot and washed with distilled water and dried
with a paper towel. A glass slide was taken and placed in a petriplate.
Distilled water or dierent buers in 100 L volumes were poured
around the middle of the glass slide with the help of micropipette. A
small pool of liquid was thus formed on the slide, on which the snail
was placed. Following this the back of its foot was gently scraped with
the help of a spatula to induce yellowish mucus secretion. After the
secretion of the mucus the snail was transferred back to its habitat. The
mucus was either transferred from the slide to a microcentrifuge tube
or the glass slide was subjected to adhesive force measurement. The
buers into which the mucus was collected were acetate buer (pH 5),
phosphate buers (pH 6 and pH 8), tris buer (pH 7 and pH 8),
glycine-NaOH buers (pH 9 and pH 10) and bicarbonate buers (pH
9 and pH 10).
In addition to above buers the mucus was also exposed to two
other solutions. These are ferric chloride solution (5 mM) in water
and sodium borohydride (NaBH4) solution (10 mM) in NaOH, pH
11.5. Eects of all of the above solutions on the adhesive property of
the mucus were studied. Mucus pH was also noted after its release and
mixing with water or buers, with the help of pH papers. Standard pH
meters could not be used for this purpose because of the small volume
and viscous nature of the mucus.
Biochemical Assay. The viscous adhesive mucus was collected in
microcentrifuge tubes. For protein extraction the method described by
Smith et al. (2002)11 was followed. Mucus samples were vortexed for
23 min and then either sonicated or heated in a water bath at 80 C
for 10 min. Sonication was carried out at 60% amplitude and 30 s pulse
rate. After this, the sample was again vortexed for 34 min and then
centrifuged for 15 min at 13000 rpm The yellow colored supernatant
was taken out in a fresh microcentrifuge tube. This supernatant is a
nonviscous liquid which was subjected to carbohydrate estimation by
orcinol-sulfuric acid method and protein estimation by bicinchoninic
acid method (Pierce Chemical Co.).
Measurement of Adhesive Force. Glass slides containing freshly
collected mucus was immediately subjected to adhesive force
measurement. The glass slide itself acted as the test surface. Another
glass slide was immediately placed on the top of the mucus on the rst

Figure 2. (a) Design of the instrument used to measure the force

required to detach the slides that were attached with snail adhesive
mucus. (b) An image of the instrument in the process of measuring
adhesive force. (Inset) Snail mucus in between two glass slides.
and picture). In this method, the glass slides, with the thin layer of glue
sandwiched between them, were placed on a small platform in the
instrument. A plastic weighing pan was attached through a pulley to
the upper slide with the help of paper clips, while the lower slide
remained xed to the instrument. Increasing weights were placed on
the plastic pan until the attachment failure. Adhesive force in Pascal
(Pa = N/m2) was manually calculated as maximum weight needed to
detach the glass slides per unit overlapping area.
We checked the performance of the above instrument with gelatin
solutions of dierent concentrations. The experiment was a
modication of the one described by Smith (2002).11 Briey, gelatin
solution (50 L) was placed on the center of a glass slide. Another
slide was placed on this solution on the rst glass slide forming a cross.
The slides were held together in position, with the gelatin solution in
between, with a clip and kept in 4 C for 23 h. The slides were then
pulled apart with the instrument and the corresponding adhesive
forces were recorded. Values of the adhesive forces were comparable
to those obtained by Smith (2002)12 (Figure 3). Nevertheless a few
limitations of the above instrument need to be mentioned. For
instance, during experiment with snail mucus, distribution of the snail

Figure 3. Adhesive load measured using our equipment () was

compared that () reported in the literature.10 Here a layer of gelatin
was used as the test material which was sandwiched between two
parallel glass plates. The tensile stress required to separate these plates
were measured. The values represent average of ve readings, the error
bar represent standard deviation.
B

DOI: 10.1021/acs.langmuir.5b03498
Langmuir XXXX, XXX, XXXXXX

Article

Langmuir

assay by Orcinol-Sulfuric acid assay. On an average 225 g of

protein and 120 g of carbohydrate per mucus sample was
obtained. However, a small part of the mucus is lost as pellet.
So above values likely to be an underestimate. Detailed
biochemical analysis of the mucus is required to understand the
contribution of the biomolecules in the adhesive mechanism.
Determination of pH of the Mucus. The eect of the
buers on the intrinsic pH of the mucus was also determined
with the help of pH papers. Table 1 shows the result of this

mucus over the surface enclosed by the two glass slides was not
uniform. Also elimination of bubbles from the enclosed portion was
not possible. Since the mucus was in a viscous liquid form, its presence
did not create any signicant gap in between the two slides. A part of
the mucus overowed out of the enclosed region and small errors may
creep in due to surface tension of this extra liquid. But attempts could
not be made to wipe o this liquid, because we could not risk
disturbing the system once the upper slide was placed on the slippery
mucus. A third source of error may come from manual increase of load
on the weighing pan. In the course of placing gradually increasing
weights on the pan, 23 min passed and during this time period the
adhesive mucus remained under strain. This may have negative eect
on adhesion and introduce error in the measured value of the adhesive
force.
Rheological Studies. Mucus samples from individual snails were
collected in four dierent liquids, acetate buer (pH 5), bicarbonate
buer (pH 10), distilled water, and FeCl3 solution, each 250 L of
volume. Immediately after collection they were subjected to frequency
scan at xed 1% amplitude in a dynamic rheometer (Anton-Paar
MCR302, Austria) in the 0.3300 rad/s range. Storage and loss
moduli of each sample were found to maintain almost constant values
with slight changes in approximately 1.517 rad/s range. Storage
modulus of each mucus sample was recorded at a xed frequency of
10.68 rad/s.

Table 1. pH of the Mucus in the Presence of Dierent

Solutions

RESULTS
Maintenance of the Snail in the Laboratory. The
present study was done with the terrestrial snail Macrochlamys
indica. If these animals are disturbed, sometimes they behave as
if they sense danger and secrete an yellow colored mucus
(Figure 1(b)). This mucus is more viscous than the colorless
trail mucus of the snail; it is sticky to touch, unlike trail mucus,
it becomes a semisolid gel with time and, after drying, leaves a
solid residue. The semisolid mass was found to be stretchable
and able to hold paper clip fragments hanging from a glass slide
(see Figure 4). This adhesive mucus was studied for its protein

buer/solution

pH of the
buer

pH of the mucus collected in the

buer

acetate buer
phosphate buer
water
tris buer
bicarbonate buer
glycine NaOH
bicarbonate buer
NaBH4
FeCl3

5
6
7.1
8
9
9
10
11.5
2.6

6.5
6.5
8.5
8.5
9
9
10.5
10.5
8.5

study. It was found that in most of the cases the pH of the

mucus-buer mixture was dierent from the buer itself. The
pH of the mucus was close to 8.5. It was the same in the
presence of distilled water, which is expected to cause least
change in the intrinsic pH of the mucus. When the mucus was
collected in acetate, tris or phosphate buers, the mixture
showed higher pH than the original buer, while in case of
NaBH4 solution (pH 11.5), the mucus caused lowering of pH.
Change of pH was not observed in case of bicarbonate or
glycine-NaOH buers (pH 9). All these results conrm that
intrinsic pH of the mucus to be mildly alkaline. It is notable that
observed adhesive strength was generally high in the presence
of alkaline buers, i.e. around pH 9 and pH10.
Adhesive Property of the Mucus under Dierent
Conditions. The snail was made to secrete adhesive mucus on
one of the two glass slides which were then attached together.
The slide contained dierent buers of 100 L volume, on
which the snail was placed and they released the mucus into the
liquid. Adhesive force was measured within minutes of
collection of the mucus. In this process the mucus was exposed
to dierent pH condition and was brought in contact with the
glass surface before it was completely cross-linked. During
natural release by the snail, the adhesive comes in contact with
its target surface instantly. In order to mimic this natural
condition we ensured that the induced adhesive mucus
secretion interacts with the test surface immediately. The
force needed to separate the slides held together with the wet
viscous mucus was measured. Since adhesives are generally less
active or completely inactive under water, it is expected that
performance of the mucus as an adhesive would be
compromised in the presence of these liquids. However,
natural habitat of snails is damp and humid. It can be assumed
that the snail may secrete this mucus on moist surfaces and yet
immediate performance of the mucus as adhesive would be
useful.
Figure 5 shows the eects of dierent buers on the mucus.
In presence of alkaline buers, glycine-NaOH and bicarbonate
(pH 9 and 10), the force needed to detach the glass slides was

Figure 4. Firmness and stretchability of the adhesive mucus: (a)

Gelled mucus can hold a paper clip hanging attached from a glass slide,
(b) Gelled mucus is stretchable.

and carbohydrate content and its adhesive property. Snails

could be maintained in the laboratory condition for about a
couple of months. During this time period adhesive mucus
could be collected from each snail 34 times.
Mucus of other species of snails and slugs were known to be
a mixture of proteins and carbohydrates. Through biochemical
assays we veried that such is also the composition in the
present case. For this, after collection, the mucus was
transferred to eppendorfs and subjected to sonication and
vortexing. After centrifugation of this mixture, an extract with
less viscosity was obtained which was subjected to protein
estimation by bicinchoninic acid (BCA) assay and carbohydrate
C

DOI: 10.1021/acs.langmuir.5b03498
Langmuir XXXX, XXX, XXXXXX

Article

Langmuir

Figure 6. (a) Dry weight and (b) Wet weight of mucus collected in
dierent conditions. The values represent average of ve samples, the
error bar represent standard error of mean. Signicant dierence of
wet weights of acetate and bicarbonate buers was found (Wilcoxon
rank sum p value 0.032).

Figure 5. Box plot of the force needed to detach two glass slides
attached by snail adhesive mucus in dierent buers. The boxes
indicate the 25th and 75th percentiles and error bars indicate the
minimum and maximum values. Spearmans rank correlation
coecient between pH value and adhesive force was 0.86, p 0.003.
The sample size ranged from 9 to16.

highest (9.8 kPa, average 6.5 kPa, median 6.8 kPa), while in the
presence of the acidic acetate buer, the average force was 2.5
kPa. Although, the average force in phosphate pH 7 was 3.3
kPa, it was again low (2.9 and 2.6 kPa, respectively) in the
presence of two dierent buers phosphate and tris, both
having pH 8.
Signicant dierence was found between the adhesive force
values at two extreme conditions, bicarbonate buer (pH 10)
and acetate buer (pH 5) (MannWhitney test p-value 5
106). There was signicant correlation between medians of
adhesive force and pH values (Spearmans rank correlation
coecient 0.86, p 0.003).
During collection of adhesive mucus a dierence was noted
in their release in dierent buers. The snail could be induced
to release yellow adhesive mucus very easily in the presence of
alkaline pH, while in acetate buer the same process needed
longer time. In order to test if this dierence actually leads to
dierence in amount of mucus release, we determined wet
weight of the mucus collected in two extreme pH and also in
the presence of distilled water and FeCl3 solution. After drying
the mucus again we determined the dry weight of the mucus.
As presented in Figure 6 there was no signicant dierence
between the average dry weights of mucus collected under
dierent conditions in general, but wet weights of mucus
collected in the bicarbonate buer was signicantly higher than
that collected in the acetate buer (p-value = 0.03175, by
Wilcoxon rank sum test). This indicates amount of total water
released with mucus in these two conditions may be dierent.
We observed a dierence in appearances of the mucus in
acidic and alkaline buers. In alkaline buer the mucus
appeared comparatively more viscous. The newly secreted
mucus, when mixed with the alkaline liquid on the slide,
incorporated water and formed a viscous liquid drop with a
large contact angle. In presence of acidic buer, the mucus was
less viscous with small contact angle and water incorporation
was less (see Figure 7).
In addition to the buers, the eect two other conditions
were also checked on the adhesive property of the mucus, these
are FeCl3 and NaBH4 solutions. From the work of Werneke et
al. (2007)6 it is known that metal dependent oxidation of lysine
residues may form carbonyl groups which causes cross-linking

Figure 7. Typical appearance of mucus providing dierent adhesive

forces. (Snails secreted mucus on glass slides containing buers of
dierent pH in 100 L volumes and the mucus was immediately mixed
with the buer). In alkaline buer the mucus incorporated all of the
water on the slide and became thick and viscous. In acidic buer the
mucus failed to incorporate all the free water.

in adhesive mucus. We exposed the mucus to FeCl3 solution on

the assumption that it may increase the cross-linking in the
mucus and increase its rmness. We, in fact, observed
qualitative change in the mucus in FeCl3 which became more
viscous and sti than the mucus in alkaline buer. All the water
on the slide was incorporated in viscous gel structure and the
slide surface became dry. The initial pH of FeCl3 solution was
2.6 which changed in contact with the mucus to 8.5 (see Table
1). Adhesive strength of this mixture was moderately high with
average value of 5.4 kPa.
The observed increase in stiness of the mucus in FeCl3 was
veried by determining its storage moduli in four dierent
conditions, acetate buer (pH 5), bicarbonate buer (pH 10),
distilled water and FeCl3 solution. Figure 8 shows that while
there were no signicant dierence among the average values of
storage moduli in acetate and bicarbonate buers and distilled
water, average value of storage moduli in FeCl3 solution was
much higher than those of other three conditions.
In contrast to metal containing acidic solution, the second
solution NaBH4 was at pH 11.5, and this was used to test the
eect of mild reducing environment on the mucus. Bradshaw et
al. (2011)13 showed that oxidative cross-linking reaction of a
slug adhesive mucus occurred through imine bond formation
between carbonyl and amino groups. In presence of NaBH4 this
bond was stabilized by reduction. In our study the mucus came
in contact with NaBH4 during its release, that is, before any
D

DOI: 10.1021/acs.langmuir.5b03498
Langmuir XXXX, XXX, XXXXXX

Article

Langmuir

secretion of the mucus.6 It was considered that if the mucus

components are collected in dierent buers before the crosslinking reactions are over, it may have a dierent eect on the
nal mucus than when the buer is added after the insoluble
polymer is formed. This type of experiment was done in the
presence of EDTA before6 but not under any other condition.
We observed that in the presence of metal salt the wet mucus
had better ability to absorb water than alkaline mucus. It
demonstrated rapid increase of viscosity and stickiness which
indicated increased cross-linking between polymers. However,
adhesive property did not increase comparatively to any great
extent. Thus, performance of the mucus as wet adhesive was
compromised as cross-linking in the mucus was greatly
increased.
In a relatively reducing environment, in the presence of
NaBH4, adhesive strength of the wet mucus was on an average
greater than that in the presence of the metal. It is known that
NaBH4 can reduce carbonyl groups to alcohols15 and thus
negatively aect cross-linking reaction. Obviously this condition
did not aect the ability of the hydrated mucus as adhesive.
Presumably, the high pH (11.5) of NaBH4 solution had positive
eect on the adhesive property, while the reducing environment may not have a large eect on intermolecular interaction
within the time period between mucus secretion and adhesive
force measurement. These results indicate that in addition to
metal dependent oxidation there are other mechanisms that can
be linked to increased adhesion. These other mechanisms may
also be related to its ability for water incorporation and
responsible for the pH dependent variations in the adhesive
strength.
Studies on natural adhesives from sea organisms have
enriched the scientic knowledge of possible mechanisms of
designing underwater adhesives. For example, Dopa side-chains
of protein adhesive have been found to be able to create strong
adhesion on wet surfaces. Also following the example of
sandcastle glue Shao and Stewart developed underwater
adhesive with polyelectrolytes which formed dense cohesive
uids. Conversion of this adhesive from liquid state to gelled
state depended on the change of pH and presence of divalent
cations.16 Barnacle cements adhesive contains hydrophobic
protein able to cause underwater adhesion.17 One of the
challenging questions in underwater adhesives is the mechanism of removal of surface water layer to create attachment.
Although gastropod snails do not come in contact with as much
water as sea animals, their natural habitat is humid. The
mechanism adopted by them to remove small amount of
surface water could be absorption of this water to form the
adhesive mucus gel.

Figure 8. Storage modulus of snail mucus collected under dierent

condition. The values represent average of n readings, the error bar
represent standard error of mean.

cross-linking reaction started. Under this condition uncrosslinked carbonyl groups would be reduced to alcohols and
prevent the cross-linking reaction.1315 We found that the
adhesive strength of the mixture was greatest in the presence of
NaBH4, which could be the eect of the high alkaline pH.
General appearance of the mucus was same as that in alkaline
buer. However, the mixture slowly lost its viscosity with time
and in the dried form its adherence to the glass surface became
weak (result not shown).

DISCUSSION
Macrochlamys indica, as a terrestrial snail, is generally
considered as a garden pest. In contrast, present study is
focused on a product of the snail that can be useful, in the long
term, as biological glue, eective in moist conditions. This
unique property of biological adhesives has been explored in
case of other molluscs such as mussels and barnacles and the
extended possibility of its use as tissue adhesive has also been
studied.2 However, there are few instances of such studies on
gastropod adhesive mucus.
In this work, we detected adhesive mucus in a snail and
investigated their ability to attach two glass surfaces together.
The mucus was subjected to dierent conditions during its
secretion. These conditions include exposure to buers of
dierent pH. We found that the adhesive property is pH
dependent; it is low at pH 5 or 6 and increases with increase of
pH. Also our results indicate that intrinsic pH of the mucus is
alkaline. Since detailed biochemical studies are not done yet the
exact reason for this alkalinity is not known; it can be assumed
there is an abundance of alkaline groups, such as amino groups,
in the protein and carbohydrate components of the mucus.
Dierences in adhesive property seem also to be related to
greater amount of water in the released mucus and its ability to
incorporate additional water, both of which were greater in case
of alkaline pH.
Metal dependent oxidation reaction is known to form
carbonyl groups which get involved in cross-linking the
polymers through imine bond formation.14 This increased
cross-linking reaction may be responsible for the increased
cohesion and lm formation. In the present study we have
investigated the eect of addition of metal salt. For this we
collected the mucus in FeCl3 solution (pH of 2.6) and studied
the adhesive property of the mixture. It is known that metal
dependent oxidation reaction is completed within seconds after

SUMMARY

We studied the ability of the mucus to attach two wet glass

slides in dierent pH and in the presence of metal salts. The
adhesive force was more in higher pH and the maximum force
needed to detach these glass slides was approximately 9 kPa.
This ability of the mucus to attach the wet glass slides was
related to its ability to incorporated water from the wet slides in
its viscous gel structure. Similar studies with other gastropods
may show even greater adhesive strength and we believe that
such studies will reveal the real merit of gastropod adhesive
mucus.
E

DOI: 10.1021/acs.langmuir.5b03498
Langmuir XXXX, XXX, XXXXXX

Article

Langmuir

(16) Hui, S.; Stewart, R. J. Biomimetic Underwater Adhesives with

Environmentally Triggered Setting Mechanisms. Adv. Mater. 2010, 22,
729733.
(17) Kamino, K. Molecular Design of Barnacle Cement in
Comparison with Those of Mussel and Tubeworm. J. Adhes. 2010,
86, 96110.

AUTHOR INFORMATION

Corresponding Author

*E-mail: aghatak@kiitbiotech.ac.in.
Notes

The authors declare no competing nancial interest.

ACKNOWLEDGMENTS
This work was supported by the nancial assistance of
Department of Science and Technology, Govt. of India with
project le no. SB/YS/LS-375/2013 and a research initiation
grant from KIIT University, Odisha, India. A.G. acknowledges
Dr. Goutam Tripathy, Institute of Physics for his help in
conducting rheological experiments, Dr. Gajinder Pal Singh,
KIIT University for his help in carrying out statistical analysis of
the data, Professor L.C. Padhy, KIIT University and Professor
Subrata Tripathi, Institute of Physics, Odisha, India, for their
valuable comments.

REFERENCES

(1) Smith, A. M. The biochemistry and mechanics of gastropod

adhesive gels. In Biological Adhesives; Smith, A. M., Callow, J. A., Eds.;
Springer-Verlag: Berlin Heidelberg, 2006; pp 167182.
(2) Silverman, H. G.; Roberto, F. F. Understanding Marine Mussel
Adhesion. Mar. Biotechnol. 2007, 9, 661681.
(3) Pawlicki, J. M.; Pease, L. B.; Pierce, C. M.; Startz, T. P.; Zhang,
Y.; Smith, A. M. The effect of molluscan glue proteins on gel
mechanics. J. Exp. Biol. 2004, 207, 11271135.
(4) Ng, T. P.; Saltin, S. H.; Davies, M. S.; Johannesson, K.; Stafford,
R.; Williams, G. A. Snails and their trails: the multiple functions of
trail-following in gastropods. Biol. Rev. Camb. Philos. Soc. 2013, 88,
683700.
(5) Smith, A. M.; Quick, T. J.; Peter, R. L., St. Differences in the
composition of adhesive and non-adhesive mucus from the limpet
Lottia limatula. Biol. Bull. 1999, 196, 3444.
(6) Werneke, S. W.; Swann, C.; Farquharson, L. A.; Hamilton, K. S.;
Smith, A. M. The role of metals in molluscan adhesive gels. J. Exp. Biol.
2007, 210, 21372145.
(7) Braun, M.; Menges, M.; Opoku, F.; Smith, A. M. The relative
contribution of calcium, zinc and oxidation-based cross-links to the
stiffness of Arion subf uscus glue. J. Exp. Biol. 2013, 216, 14751483.
(8) Blanford, W. T.; Godwin-Austen, H. H. Mollusca, Testacellidae
and Zonitidae. The Fauna of British India; Taylor and Francis: London,
1908; pp 9597.
(9) Jahan, M. S.; Kulsum, M. U.; Rahman, M. R.; Sarker, M. M.;
Pramanik, M. N. Comparative Ecology of Macrochlamys indica
Godwin-Austin and Macrochlamys opiparus Godwin-Austin (Stylommatophora: Ariophantidae). J. Ecobiol. 2002, 14, 307318 (India).
(10) El-Alfy, N. Z.; Al-Ali, K. A.; Abdel-Rehim, A. H. Karyotype,
meiosis and sperm formation in the land snail Macrochlamys indica.
Cytologia 1994, 14, 122128.
(11) Smith, A. M.; Morin, M. C. Biochemical differences between
trail mucus and adhesive mucus from marsh periwinkle snails. Biol.
Bull. 2002, 203, 338346.
(12) Smith, A. M. The structure and function of adhesive gels from
invertebrates. Integr. Comp. Biol. 2002, 42, 11641171.
(13) Bradshaw, A.; Salt, M.; Bell, A.; Zeitler, M.; Litra, N.; Smith, A.
M. Cross-linking by protein oxidation in the rapidly setting gel-based
glues of slugs. J. Exp. Biol. 2011, 214, 16991706.
(14) Requena, J. R.; Chao, C. C.; Levine, R. L.; Stadtman, E. R.
Glutamic and aminoadipic semialdehydes are the main carbonyl
products of metal-catalyzed oxidation of proteins. Proc. Natl. Acad. Sci.
U. S. A. 2001, 98, 6974.
(15) Dalle-Donne, I.; Rossi, R.; Giustarini, D.; Milzani, A.; Colombo,
R. Protein carbonyl groups as biomarkers of oxidative stress. Clin.
Chim. Acta 2003, 329, 2338.
F

DOI: 10.1021/acs.langmuir.5b03498
Langmuir XXXX, XXX, XXXXXX