Journal of the Marine Biological Association of the United Kingdom, 2016, 96(6), 1191 – 1199.

# Marine Biological Association of the United Kingdom, 2015

doi:10.1017/S0025315415000715

The effects of salinity on the density, shell

size and survival of a mangrove gastropod:
laboratory and field evidence
rafaela camargo maia1 and ricardo coutinho2
1
Laboratório de Ecologia de Manguezais, Instituto Federal de Educação, Ciência e Tecnologia do Ceará, Campus Acaraú, Avenida
Desembargador Armando de Sales Louzada, s/n, CEP 62580-000 Acaraú – CE, Brazil, 2Departamento de Oceanografia, Divisão de
Biotecnologia Marinha, Instituto de Estudos do Mar Almirante Paulo Moreira, Arraial do Cabo, RJ, CEP: 28930-000, Brazil

The macro-detritivore gastropod Melampus coffeus plays an important role in energy transfer in neotropical mangroves and,
because it consumes tree leaves, it may be a potential ecological indicator of degraded mangrove areas. The objective of this
study was to analyse the spatial-temporal distribution and population dynamics parameters of M. coffeus in mangroves and
correlate environmental variables with population density, shell morphology and survival. Samples were collected monthly in
two mangrove forests with different salinities, located on the north-eastern coast of Brazil. Height, width and aperture height
of the animals’ shell were measured. The effects of salinity on population density and size distribution in M. coffeus were
evaluated in field and laboratory experiments. The results showed that populations of M. coffeus present low density and
are composed of large individuals during the dry season in both mangrove forests. These populations are denser and show
predominance of small individuals during the rainy season when salinity decreases. The results obtained in the experiments
confirm the observations in the field. Animals at extreme sizes (small and large) subjected to different salinity treatments over
a moderate period showed higher mortality rates than individuals of intermediate size.

Keywords: Growth, Melampus coffeus, Pulmonata, tropical estuary, shell morphology

Submitted 3 December 2013; accepted 22 April 2014; first published online 28 July 2015

INTRODUCTION the population dynamics of the Ellobiidae family and tidal

The gastropod Melampus coffeus (Linnaeus, 1758) belongs to a
primitive group of mainly marine pulmonate molluscs com-
monly found at the upper levels of the intertidal zone of man-
groves in the Atlantic Ocean (Martins, 1996a, b). The
regulation of snail populations in mangroves or tropical estuar-
ies is subjected to processes that are density dependent and
independent such as competition and predation, and tempera-
ture, rainfall, and salinity, respectively (Roach & Lim, 2000;
Maia et al., 2012). In the case of Melampus, predation and
food competition seem to be the main density-dependent regu-
latory factors (Joyce & Weisberg, 1986; Lee & Silliman, 2006),
whereas seasonal fluctuations in water salinity and temperature,
which can lead to significant mortality of young animals and
eggs, are the most important density-independent factors
(Apley, 1970; Price, 1980; McMahon & Russell-Hunter, 1981;
Maia et al., 2012).
Melampus species present high densities in areas with high
salinities (Price, 1980; McMahon & Russell-Hunter, 1981; Fell
& Williams, 1985; Burnham & Fell, 1989). However, observed
variation in the densities of M. coffeus did not corroborate the
patterns recorded for other species in Melampus; instead, high
densities were detected in areas with low salinities (Maia &
Coutinho, 2013). Other studies show a relationship between
Corresponding author:
R.C. Maia
Email: rafaelamaia@ifce.edu.br
levels. Some species in Melampus can display an elaborate suc-
cession of temporal adjustments in their reproductive and
embryonic development, which correspond to submersion
during spring tides (Apley, 1970; Russell-Hunter et al., 1972;
Spelke et al., 1995). Maia et al. (2012) show that, in
Melampus coffeus, no reproductive activity occurs during
the dry season, after which, increasing rainfall and reduced
salinity result in very dense populations of predominantly
small individuals.
As a macro-detritivore mollusc, M. coffeus plays an import-
ant role in energy transfer in neotropical mangroves and its
distribution and abundance appear to be conditioned to the
presence of trees in the mangrove, which would represent
both a condition and a resource (Joyce & Weisberg, 1986;
Lee & Silliman, 2006). This snail is an air breather and, there-
fore, migrates to the upper parts of the mangrove vegetation
during high tides to avoid drowning (Maia & Tanaka,
2007). However, M. coffeus feeds on decaying leaf litter on
the sediment surface (Proffitt et al., 1993; Proffitt & Devlin,
2005). Maia & Coutinho (2013) indicate congruent distribu-
tion patterns of mangrove trees and M. coffeus on the north-
eastern Brazilian coast, that is, the greatest availability of food
resources and shelter, provided by the mangrove vegetation,
correlated with the greatest snail abundance. This result indi-
cates that M. coffeus may be a potential ecological indicator of
mangrove areas degraded by deforestation.
The biology and ecology of Ellobiidae is described in
studies such as those on larval development, life history and

1191

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 1192 rafaela camargo maia and ricardo coutinho
distribution patterns (Apley, 1970; Russell-Hunter et al., 1972,
Spelke et al., 1995). However, little is known about the biology
and ecology of M. coffeus (Proffitt et al., 1993; Proffitt &
Devlin, 2005). The use of this species as a bioindicator requires
knowledge not only of species or population characteristics
such as specimen size, distribution and density, but also
knowledge of how environmental conditions in mangrove
estuaries (salinity, vegetation cover) affect M. coffeus over
time.
Thus, the goal of this study was to analyse the spatial and
temporal distribution of Melampus coffeus in two mangroves,
with different salinities on the north-eastern coast of Brazil
(Ceará State). In addition, population dynamics parameters
such as survival, population density and shell size (shell
height – ventral margin of aperture to apex) were correlated
with environmental variables (temperature, salinity and rela-
tive humidity). Our working hypothesis was that population
density and individual shell morphology are influenced by sal-
inity (personal observation). Thus, we expected decreases in
population density and increases in the shell size in the
season with high salinities.
MATERIALS AND METHODS
Study area
In order to evaluate the spatial and temporal distribution of
M. coffeus and the variables that are correlated with its popu-
lation dynamics, we selected two mangrove forests with differ-
ent salinity levels and a similar vegetated area in the county
of Acaraú, Ceará State, Brazil. The first is located in the
middle section of the Acaraú estuary (Acaraú River man-
grove), a less saline area with fluvial influence (02850′ 59′′ S,
40807′ 41′′ W) with seasonal fluctuations in water salinity
between 0 and 44 ppt. The second mangrove forest is
located on the Arpoeiras Beach (Arpoeiras Beach mangrove),
a more saline environment due to its proximity to the sea
(02850′ 17′′ S 40804′ 56′′ W), with salinities between 10 and
51 ppt.
The coastal zone of Ceará is included in segment IV of the
Brazilian coastal division proposed by Schaeffer-Novelli et al.
(1990). Rainfall is low in this area leading to an accumulation
of salt due to high rates of evaporation in the estuarine areas,
whereas the temperature remains constant throughout the
year. In both study areas, the mangrove species Rhizophora
mangle Linnaeus (Rhizophoraceae) and Avicennia germinans
(Linnaeus) Stearn were equally dominant and there was no
evidence of a clear zonation pattern (Maia & Coutinho, 2012).
Methodology
field procedures
Each month between March 2007 and February 2009, five plots
(1 m2 each) were randomly selected in each mangrove forest.
All specimens found within each plot were counted and mea-
sured for shell height, width and aperture height with a
caliper (precision ¼ 0.1 mm) and returned to their habitats.
The monthly average population density was calculated for
each mangrove forest. Water, air and sediment temperatures,
and relative humidity were measured with a thermo-
hygrometer; salinity was measured with a refractometer.
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Rainfall data (monthly accumulated total – mm) were obtained
from FUNCEME – Fundação Cearense de Meteorologia e
Recursos Hı́dricos (http://www.funceme.br/). Sampling was
conducted during daylight spring tides; tides were identified
according to the tide tables published on the website by the
DHN – Diretoria de Hidrografia e Navegação da Marinha do
Brasil (http://www.dhn.mar.mil.br/).
The two-way Analysis of Variance test was used to investi-
gate differences in mean density or shell size (height, width
and aperture height) among the different sampling months
and study areas (Underwood, 1997). Tukey’s multiple com-
parison test was used when differences were observed.
The Spearman correlation analysis was used to correlate
density and shell height with environmental parameters (air,
water and sediment temperature, humidity, salinity and rain-
fall). Other size-variables (width and aperture height) were
not used in this analysis because they were highly correlated
(R ≥ 0.95). Correlation analyses were conducted between
abiotic variables in the studied mangroves.
Size-frequency distribution of M. coffeus (shell height) was
determined according to Sturges’ formula: Vi ¼ A/K; where:
Vi ¼ class interval, A ¼ amplitude of the (Lt) variable
(Max –Min), K ¼ number of classes calculated by: 1 + 3.32
log n where ‘n’ is the total number of individuals. This calcu-
lation resulted in an estimated size class interval of 2 mm for
the Arpoeiras Beach mangrove and 1.5 mm for the Acaraú
River mangrove. These data were used for the determination
of population parameters such as growth, recruitment and
survival. All results are presented as mean + standard
deviation (SD).
Experiments
A series of experiments were conducted to test the effects of
salinity on M. coffeus population density and individual size
distribution.
experiment i -- effects of salinity on
m. coffeus
Eight M. coffeus specimens with similar shell sizes (height ¼
15 + 1 mm, and average weight ¼ 0.95427 + 0.1514 g)
were collected at both study areas. These animals were taken
to the laboratory, placed in trays (302 × 208 × 63 mm) at a
density similar to that found in their natural habitat and
sprayed with water at different salinities: Treatment # 1 ¼
0 ppt, Treatment # 2 ¼ 32 ppt, and Treatment # 3 ¼ 48 ppt.
Sediment from the source mangrove and yellow mangrove
leaves were added to the trays to provide abundant food.
The trays were watered daily for 2 months using the salinity
concentrations of each treatment. Shell height (mm) was mea-
sured and individual snails were weighed weekly (precision ¼
0.0001 g).
The effects of salinity on the survival of M. coffeus (%) were
assessed through a two-way Analysis of Variance (Salinity
treatments and Time in weeks), where Treatment # 1 ¼
0 ppt, Treatment # 2 ¼ 32 ppt, and Treatment # 3 ¼ 48 ppt
(salinity), were applied to each study site. Differences in sur-
vival between both study areas were assessed through a
one-way Analysis of Variance (Underwood, 1997). Shell
growth (final height – initial height) and loss of body
weight (final weight – initial weight) were assessed through
a two-way Analysis of Variance among the treatments and
 effects of salinity on a mangrove gastropod 1193

between the study areas. Tukey’s multiple comparison test was rainfall in both Arpoeiras Beach (r ¼ 20.890; P , 0.05) and
used when significant differences were observed. Acarau River (r ¼ 20.825, P , 0.05).

experiment ii -- effects of salinity on

m. coffeus of different sizes Spatial and temporal distribution of M. coffeus
This experiment was performed under the same conditions
Significant differences in the population density of M. coffeus
used in Experiment 1 except using small snails (shell
were observed among different months at Arpoeiras Beach
height ¼ 12 + 1 mm and average weight ¼ 0.46765 +
mangrove and Acaraú River mangrove (Table 1)
0.1607 g) and large snails (shell height 20 + 1 mm and
(Figure 1A). Variation in density between the studied
average weight ¼ 1.85177 + 0.25335 g) to evaluate the influ-
months was similar in both study areas. According to
ence of salinity on different size classes.
Tukey’s test, the population density of M. coffeus in July
The effects of salinity on the survival of M. coffeus was
2007 was significantly different from densities in January
assessed through a two-way Analysis of Variance (Salinity
and November 2008 in the Arpoeiras Beach mangrove; the
treatments and Time in weeks) for each size class (small and
density in March 2007 was different from densities in
large snails). Differences in survival between both size
October 2007, and in October and December 2008, in the
classes were assessed through a one-way Analysis of
Acaraú River mangrove. All other months were statistically
Variance (Underwood, 1997). Shell growth (final height –
similar. Thus, the sampled mangroves forest tend to present
initial height) and loss of body weight (final weight – initial
lower densities in months with low rainfall and high salinity.
weight) was assessed through a two-way Analysis of
No significant differences in density occurred between the
Variance (Salinity treatments and Time in weeks). Tukey’s
study areas (Table 1); M. coffeus densities were similar at
multiple comparison test was used when significant differ-
both study areas (Figure 1A).
ences were observed.
The size of M. coffeus (Figure 1B) was significantly different
among months at study areas, considering shell height, width
experiment iii -- transplant and aperture height (Table 1). At Arpoeiras Beach mangrove,
Additionally, an experimental transplant was conducted over according to Tukey’s multiple comparison test, height, width
3 months to evaluate the effects of different salinity concentra- and aperture height were similar in: (1) March, April,
tions on shell height in their natural environment. Thirty indi- June and July 2007, July, August, September, October,
viduals randomly collected in the Acaraú River mangrove November and December 2008, and January 2009; (2) May,
were transferred to the Arpoeiras Beach mangrove, whereas July, August, October, November and December 2007; (3)
30 specimens collected in Arpoeiras Beach mangrove September 2007, March and June 2008; and (4) January,
were transferred to the Acaraú River mangrove. Because
M. coffeus migrates vertically through the course of a day,
Table 1. Results of two-way Analysis of Variance comparing density and
feeding on the sediment during low tides and climbing up shell size (height, width and aperture height, mm) of Melampus coffeus in
trees during high tides (Proffitt & Devlin, 2005; Maia & the sampling months (May 2007 – January 2009) at the Arpoeiras Beach
Tanaka, 2007), the animals were kept in suitable containers, and Acaraú River mangrove forests, Ceará state, Brazil.
measuring 1 m in height and 25 cm2, with open tops to
allow leaves to enter. These containers, three in each area, Effect Density
were built with a nylon screen on a wooden frame of man- SS DF MS F P
grove branches. All animals were numbered with enamel
paint, and had their shell height (mm) measured once a Study sites 2.40 1 2.40 0.0207 0.88570ns
month. A paired Student’s t-test was used to evaluate shell Sampling months 7559.98 23 328.69 2.8374 0.00005∗
Study sites∗ sampling 1968 23 85.57 0.7386 0.80165ns
growth in the transplanted individuals.
months
Error 22241.6 192 115.84
Shell height
RESULTS Study sites 1082 1 1082 140.51 ,0.00001∗
Sampling months 2280 23 99.1 12.87 ,0.00001∗
Study sites∗ sampling 603.8 23 27.6 3.59 ,0.00001∗
Environmental variables in the study areas months
The temperature data from air, water and sediment were Error 20114.8 2612 7.7
similar in both mangrove forests throughout the study. Shell width
However, the relative air humidity varied in both study Study sites 641.4 1 641.4 72.74 ,0.00001∗
areas, between different months, with a 27% minimum Sampling months 1133.1 23 49.3 5.59 ,0.00001∗
humidity registered in the Acaraú River mangrove in Study sites∗ sampling 368.3 23 16 1.82 0.010∗
October 2008, and an 82% maximum humidity recorded in months
Error 23034.7 2612 8.8
Arpoeiras Beach mangrove in March 2007. The total accumu-
lated rainfall ranged between 0 and 477.5 mm during this Shell aperture height
study (recorded in April 2008). According to data provided Study sites 1036.4 1 1036.4 136.78 ,0.00001∗
Sampling months 1902.9 23 82.7 10.92 ,0.00001∗
by FUNCEME, three rainy periods occurred: from March to
Study sites∗ sampling 544.8 23 23.7 3.13 0.00001∗
April 2007, from January to May 2008, and from January
months
to February 2009. Two dry periods were observed from May Error 19792.6 2612 7.6
to December 2007 and from June to December 2008. The ana-
lysis indicated a significant correlation between salinity and ns
No significant variation; ∗ Significant variation (P , 0.05).

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 1194 rafaela camargo maia and ricardo coutinho

Fig. 1. (A) Density (individual m22, mean + SD) and (B) Shell height (mm) of M. coffeus sampled between March 2007 and January 2009 at the Arpoeiras Beach
and Acaraú mangrove forests, Ceará state, Brazil.

February and April 2008. At Acaraú River mangrove, accord-
ing to Tukey’s multiple comparison test, the following groups
of months were similar among themselves and distinct from
the others: (1) March 2007, November 2008 and February
2009; and (2) July, August, September and October 2007.
Temporal patterns in shell size at the Arpoeiras Beach
mangrove and the Acaraú River mangrove are similar
(Figure 1B). In periods of drought, M. coffeus populations
tended to have a low density and were composed of large
snails. When rainfall increased and salinity levels fell, the
populations became denser and were dominated by small
individuals. Even though the change in shell size over
months was similar in both study areas, the average shell
size in the Arpoeiras Beach and in the Acaraú River
mangroves was significantly different (Table 1). Thus, we
observed that animals collected on the Arpoeiras Beach man-
grove had significantly larger shells than those sampled in the
Acaraú River mangrove (Figure 1B).
The results, in both study areas, of the correlation analysis
between environmental variables (temperature, rainfall, salin-
ity and relative humidity), densities and shell height of M.
coffeus confirmed the patterns observed in the graphs, i.e. sig-
nificant correlation between these factors and salinity (P ,
0.05) (Table 2). At Arpoeiras Beach and Acaraú River man-
groves, the analysis indicates a positive correlation between
M. coffeus densities and rainfall (Arpoeiras Beach mangrove:
R ¼ 0.583; Acaraú River mangrove: R ¼ 0.617); and a nega-
tive correlation with salinity (Arpoeiras Beach mangrove:

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 effects of salinity on a mangrove gastropod 1195

Table 2. Correlations between Melampus coffeus density, shell height and and therefore, more size classes were represented in those
environmental variables at the Arpoeiras Beach and Acaraú River man- months (from January to June). However, a few animals
grove forests, Ceará state, Brazil. reached sizes less than 7.6 mm. In the dry months, the
Variable Arpoeiras Beach Acaraú River number of juveniles was lower and the number of adults
mangrove mangrove increased in the population, thus, a reduction in the number
of size classes was observed. Intermediate sized individuals
Density Shell Density Shell were observed throughout the period, with those ranging
height height from 13.6 to 21.6 mm being most frequent. Individuals
Density (ind m22) – 20.230ns – 20.694∗ larger than 23.6 mm were observed only in September 2007,
Shell height (mm) 20.200ns – 20.694∗ – and February, March and October 2008. Only one individual
Air temperature (8C) 20.127ns 0.054ns 0.389ns 20.228ns was found in each of these months.
Water temperature (8C) 0.154ns 20.013ns 0.416∗ 20.405∗ Compared with the Arpoeiras Beach mangrove, the Acaraú
Sediment temperature 0.562ns 20.176ns 0.131ns 20.236ns River mangrove had smaller individuals, with the first size
(8C) class ranging from 4.9 to 6.4 mm. Few animals reached sizes
Humidity (%) 0.375ns 20.072ns 0.068ns 0.068ns less than 6.4 mm or larger than 21.4 mm. Intermediate
Salinity (ppt) 20.584∗ 0.326ns 20.532∗
0.665∗ size classes, ranging from 10.9 to 18.4 mm, were more fre-
Rainfall (mm) 0.583∗ 20.203ns 0.617∗ 20.595∗
quent than other classes. In general, the size distribution pat-
ns
No significant variation; ∗ Significant variation (df: 22; P , 0.05). terns in the different months were similar in both habitats
with larger individuals in the dry season and smaller indivi-
duals in the rainy season when salinity decreases.
R ¼20.584; Acaraú River mangrove: R ¼ 20.532). The shell
height of M. coffeus were correlated with the same factors at
Acaraú River mangrove (Shell height and salinity: R ¼ Salinity experiment
0.665; Shell height and rainfall: R ¼ 20.595).
experiment i -- effects of salinity on
m. coffeus
size-frequency distribution of m. coffeus The survival of individuals (shell height ¼ 15 + 1 mm) col-
Melampus coffeus shell height frequency distributions at lected at Arpoeiras Beach mangrove and subjected to treat-
Arpoeiras Beach mangrove, indicate that individuals smaller ments with different salinities varied over time (Table 3).
than 9.6 mm were more frequent during the rainy season No deaths were observed until the fourth week in any treat-
ment, after which survival began to decrease in the higher sal-
inity treatment after 4 weeks. In intermediate salinity
Table 3. Results of two-way Analysis of Variance comparing survival (%) conditions, only one individual died in the eighth week; all
of Melampus coffeus in salinity treatments (0, 32, 48) over time (weeks) animals survived at zero salinity (Table 4). Moreover, signifi-
between Arpoeiras Beach and Acaraú River mangrove forests (Ceará cant differences in survival rates were observed in animals col-
state, Brazil) and between small (shell height ¼ 12 + 1 mm) and large lected in the Acaraú River mangrove subjected to treatments
(shell height ¼ 20 + 1 mm) snails. with different salinities over time (Table 3). In this case,
deaths were seen in all treatments after the sixth week; the
Experiment I – Effects of salinity on M. coffeus survival
lowest rates of survival were observed in the 48 ppt salinity
Effect SQ GL QM F P concentration treatment (Table 4). Nevertheless, no signifi-
cant differences in survival rates were observed between the
Arpoeiras Beach mangrove
two study areas (F1,46 ¼ 1.1329, P ¼ 0.2927).
Test weeks 4264.3 7 609.2 1.94 0.137ns
Treatment 1341.1 2 670.6 2.14 0.155ns No significant differences were observed in shell growth in
Test weeks∗ Treatment 208600.3 1 208600.3 665.54 ,0.001∗ different salinity treatments (Table 5) when comparing indivi-
Error 4388.0 14 313.4 duals from the Arpoeiras Beach mangrove and the Acaraú
Acaraú River mangrove River mangrove. However, these same animals showed a sig-
Test weeks 17994.8 7 2570.7 4.645 0.007∗ nificant treatment- and site-dependent weight loss (Table 5).
Treatments 2460.9 2 1230.5 2.224 0.145ns Weight loss was lower in snails collected in the Acaraú
Test weeks∗ Treatment 170859.4 1 170859.4 308.75 ,0.001∗ River mangrove and greatest in animals subjected to
Error 7747.4 14 553.4 Treatment #3 (salinity ¼ 48 ppt) (Figure 2A). Weight loss
was similar in the other two treatments (salinity ¼ 0 and
Experiment II – Effects of salinity on M. coffeus in different size classes
32 ppt) (Tukey’s test, Figure 2A).
Large size
Test weeks 11744.8 2 2349.8 3.068 0.0618ns experiment ii -- effects of salinity on
Treatment 3489.6 2 1744.8 2.278 0.1529ns m. coffeus of different sizes
Test weeks∗ Treatment 11828.0 1 11828.0 155.2 ,0.001∗ The second experiment using different size classes of
Error 7656.3 10 765.6 M. coffeus lasted 6 weeks because all animals were dead after
Small size this time. There were no significant differences in survival
Test weeks 9001.7 5 1800.3 2.1649 0.1397ns
rates between small (shell size ¼ 12 + 1 mm) and large
Treatment 3871.5 2 1935.8 2.3278 0.1479ns
Test weeks∗ Treatment 127092.0 1 127092.0 152.82 ,0.001∗
(20 + 1 mm) M. coffeus snails among treatments or over
Error 8316.0 10 831.6 time (Table 3). No significant differences in survival were
observed between size classes (F1,34 ¼ 0.05356, P ¼ 0.81836).
ns
No significant variation; ∗ Significant variation (P , 0.05). No deaths were recorded in any of the size classes in

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 1196 rafaela camargo maia and ricardo coutinho

Table 4. Survival (%) of small (shell height ¼ 12 + 1 mm) and large (shell height ¼ 20 + 1 mm) Melampus coffeus from mangrove forests at Arpoeiras
Beach and Acaraú River, Ceará state, Brazil, subjected to different salinities (0, 32, 48) over 8 weeks.

Week Salinity 0 Salinity 32 Salinity 48

Site Shell size Site Shell size Site Shell size

Arpoeiras Acaraú Large Small Arpoeiras Acaraú Large Small Arpoeiras Acaraú Large Small
Beach River Beach River Beach River

1 100 100 100 100 100 100 100 100 100 100 100 100
2 100 100 100 100 100 100 100 100 100 100 100 100
3 100 100 100 100 100 100 100 100 100 100 100 100
4 100 100 100 100 100 100 87.50 100 100 100 100 87.50
5 100 100 100 100 100 100 62.50 100 87.50 100 0 0
6 100 100 100 100 100 100 12.50 25 87.50 100 0 0
7 100 100 100 100 100 37.50 0 0 87.50 0 0 0
8 100 87.50 100 100 75 0 0 0 0 0 0 0

Treatment #1 (Table 4). No deaths occurred until the third
week in the other two treatments. Treatment #3 had neither
large- nor small-sized survivors in weeks five and six (Table 4).
Shell growth was similar between large and small indivi-
duals in this treatment (Table 5). However, the same
animals presented significant weight loss differences between
different treatments (Table 5). Weight loss was greater in
large animals than in small ones, especially in the intermediate
salinity treatment (Tukey’s test, Figure 2B).
experiment iii -- transplant
The animals collected in the Acaraú River and transplanted to
the Arpoeiras Beach had an average shell height of
15.5 mm + 1.7, whereas the snails sampled at the beach
and taken to the estuary had an average shell height of
14.9 mm + 1.4. It was not possible to consider all the snails

Table 5. Results of two-way Analysis of Variance comparing shell growth

(mm) and weight loss (g) of Melampus coffeus in salinity treatments (0, 32,
48) at Arpoeiras Beach and Acaraú River mangrove forests, Ceará state,
Brazil and between small (shell height ¼ 12 + 1 mm) and large (shell
height 20 + 1 mm) snails.

Experiment I – Effects of salinity on M. coffeus

Effect SQ GL QM F P

Shell growth
Treatment 0.00406 2 0.00203 0.2650 0.7684ns
Study site 0.01576 1 0.01576 1.9527 0.16929ns
Treatment∗ Study site 0.03346 2 0.01673 2.1834 0.12527ns
Error 0.3553 42 0.00807
Weight loss
Treatment 0.02807 2 0.014036 4.1188 ,0.001∗
Study site 0.049075 1 0.049075 14.3771 0.0004∗
Treatment∗ Study site 0.00125 2 0.00062 0.1836 0.023∗
Error 0.144618 42 0.00341

Experiment II – Effects of salinity on M. coffeus in different size classes

Shell growth
Treatment 0.0015 2 0.00007 0.0191 0.9810ns
Size classes 0.0002 1 0.0002 0.0517 0.8212ns
Treatment∗ Size classes 0.009 2 0.0045 1.1175 0.33661ns
Error 0.1692 42 0.00402
Weight loss
Treatment 0.03696 2 2.11545 100.777 ,0.001∗
Size classes 0.86328 1 0.86328 41.1257 ,0.001∗
Treatment∗ Size classes 0.30861 2 0.15430 7.3510 0.002∗ Fig. 2. Weight loss (g, mean + SD) of Melampus coffeus in salinity treatments
Error 0.88163 42 0.02099 (0, 32, 48) between (A) Arpoeiras Beach and Acaraú River mangrove forests,
Ceará state, Brazil and (B) between small (12 + 1 mm) and large (20 +
ns
No significant variation; ∗ Significant variation (P , 0.05). 1 mm) snails.

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used in the experiment in the statistical analysis because some
individuals were lost during the 3 months of experimentation.
Significant differences were observed in the average growth of
these animals in the experimental time at both study areas.
Animals collected from the Arpoeiras Beach mangrove and
transplanted to the Acaraú River grew 1.05 mm + 0.6 from
January to February, and 1.71 mm + 0.8, from February to
March (t ¼ 25.49389; gl ¼ 22; P ¼ 0.00016). The animals
taken from the Acaraú River mangrove and transplanted to
the Arpoeiras Beach mangrove grew 0.78 mm + 0.76 from
the first to the second month, and 1.2 mm + 0.9 from the
second to the third month (t ¼ 26.14707; gl ¼ 15; P ¼
0.000019). However, the growth rate of individuals from the
Arpoeiras Beach mangrove and transferred to Acaraú River
mangrove did not differ in comparison to those from the
Acaraú River mangrove and transferred to the Arpoeiras
Beach mangrove (t ¼ 1.6302; gl ¼ 37; P ¼ 0.111).
DISCUSSION
Salinity and rainfall appear to be associated with the density of
M. coffeus populations in both study areas and their shell size
at Arpoeiras Beach mangrove. Populations with lower density
and composed primarily of large adults occurred in the dry
season at both study areas. During the rainy season, when sal-
inity decreases, the populations were denser and small indivi-
duals predominated. Our results agree with those of other
studies that indicate that salinity is one of the main factors
determining the distribution and abundance of species in
the Melampus genus, which is found at higher densities in
areas of high salinity in estuaries (Kerwin, 1972; Fell &
Williams, 1985; Burnham & Fell, 1989; Martins, 2001; Maia
et al., 2012).
We found that seasonal variation in density and shell size
was similar in both study areas indicating that temporal vari-
ation appears more relevant. However, in general, M. coffeus
shell height was greater in the Arpoeiras Beach mangrove
than in the Acaraú River mangrove. Shell size differences in
Melampus snails and other pulmonate molluscs from neigh-
bouring regions have been reported in other studies (Fell
et al., 1991; Spelke et al., 1995; Tablado & Gappa, 2001), sug-
gesting that the differences observed could be the result of
quantitative and qualitative variation in food resources at
the two study areas in the present study, although there is
no difference in the vegetation type and tree density in both
mangroves studied (Maia & Coutinho, 2012). Melampus
coffeus is a macrodetritivore that feeds on plant debris, prefer-
ably mangrove leaves (Proffitt et al., 1993; Proffitt & Devlin,
2005). This feeding habit makes the vegetation an important
component in the population dynamics of this species.
Moreover, salinity stress, in synergy with tides, causes the
loss of leaves, which affects flows of carbon and nutrients in
estuarine mangroves exposing these systems to potential
losses by leaching and transport (Lugo et al., 1988). Thus,
net productivity, litter production and export of organic
matter are stronger in non-estuarine mangroves, which may
lead to differences in food resources available to M. coffeus
(Maia & Coutinho, 2013). and consequently larger body
sizes at Arpoeiras Beach, where the food may be more
abundant.
In the present study, the growth rate of individuals from
the Arpoeiras Beach mangrove that were transferred to the
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effects of salinity on a mangrove gastropod 1197
Acaraú River mangrove did not differ in comparison to
those from the Acaraú River mangrove transferred to the
Arpoeiras Beach mangrove. However, M. coffeus snails
appeared to grow more slowly when transplanted from the
Acaraú River mangrove to the Arpoeiras Beach mangrove,
thus, environmental conditions in the animals’ native area
may prevail over others. Field experiments involving recipro-
cal transplant discriminating between phenotypic and geno-
typic variability that may lead to morphological differences
have been suggested (Tablado & Gappa, 2001). These
authors proposed that morphometric differences observed in
the different habitats are the result of differential growth
rates in response to environmental pressures as well as food
availability.
The analysis of size-frequency distribution of M. coffeus
showed there were few very small individuals present in
both mangroves. The existence of microhabitats that provide
refuge from predation, and the limiting conditions of the
supralittoral zone may cause high mortality rates in young
individuals (Tablado & Gappa, 2001). Microhabitats were
not sampled in this study. However, we observed that
during low tides, when the snails are found on the mangrove
sediment, some individuals occupied more humid areas than
dry ones, such as mangrove roots encrusted with oysters, crab
burrows, the base of low vegetation, or small depressions on
tree root surfaces. This pattern has also been reported for
other mangrove gastropod species such as Bembicium
auratum Quoy & Gaimard, 1834 (Underwood & Barrett,
1990), Salinator solida Martens 1878 (Roach & Lim, 2000)
and Littoraria angulifera (Tanaka & Maia, 2006).
Herjanto & Thomas (1995) have also observed seasonal
differences in distinct cohorts of M. coffeus in Bermuda man-
groves where the authors report that summer populations
were dominated by juveniles indicating that recruitment
occurred in spring or early summer. Additionally, other pul-
monate gastropods exhibit temporal patterns in their life
cycle and population dynamics. Staikou et al. (1990) and
Staikou (1998) observed increased growth rates in
Bradybaena fruticum Müller, 1774 and Cepaea vindobonensis
Férussac, 1821 during the spring and early summer, respect-
ively, with their reproductive period occurring in early
summer. These authors suggested that the observed pattern
might be due to more favourable conditions during these
periods. In the present study, larger sized shells were clearly
observed during drier periods. We believe that the combin-
ation of low rainfall and high salinity during the dry season
represents a limiting factor to M. coffeus in the Acaraú
region study area, and therefore, adults in this area invest in
growth and interrupt their reproductive cycle, whereas most
young individuals and those belonging to larger size classes,
possibly the oldest, cannot withstand a prolonged dry
period. The results obtained in the experiments confirm this
assumption. Animals at extreme sizes (small and large) sub-
jected to different salinity treatments over a moderate
period showed higher mortality rates than individuals of inter-
mediate size.
Moreover, small individuals have a large surface area to
volume ratio and may lose water rapidly in the dry season
whereas larger (older) individuals may have difficulty regulat-
ing water balance (McMahon & Russell – Hunter, 1981;
Iacarella & Helmuth, 2011, 2012) Thus, younger larger indivi-
duals may have higher survival in the dry season due to a
healthy physiology and lower rates of water loss through
 1198 rafaela camargo maia and ricardo coutinho
evaporation (Britton, 1992; McMahon, 2001; Iacarella &
Helmuth, 2011, 2012).
Further studies are necessary to elucidate the processes
involved in this gastropod’s life cycle and factors that might
influence the population dynamics of this species. However,
the results from this study may contribute to the use of
Melampus coffeus, an important macro-detritivore gastropod,
as an ecological indicator of mangrove areas undergoing
degradation.
ACKNOWLEDGEMENTS
We are grateful to J.N. Maia and P.S. Ribeiro for field assist-
ance, and to A. Bismark Vasconcelos for the English style
review.
FINANCIAL SUPPORT
The Universidade Federal Fluminense and the Conselho
Nacional de Desenvolvimento Cientı́fico e Tecnológico –
CNPq provided a scholarship to the first author.
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Correspondence should be addressed to:
R.C. Maia
Laboratório de Ecologia de Manguezais, Instituto Federal de
Educação, Ciência e Tecnologia do Ceará, Campus Acaraú,
Avenida Desembargador Armando de Sales Louzada, s/n,
CEP 62580-000, Acaraú – CE, Brazil
Email: rafaelamaia@ifce.edu.br