Marine Biology (2006) 148: 479–488
DOI 10.1007/s00227-005-0097-z
R ES E AR C H A RT I C L E
E. E. G. Clavico Æ G. Muricy Æ B. A. P. da Gama
D. Batista Æ C. R. R. Ventura Æ R. C. Pereira
Ecological roles of natural products from the marine sponge
Geodia corticostylifera
Received: 17 December 2004 / Accepted: 3 August 2005 / Published online: 29 September 2005

Springer-Verlag 2005
Abstract In the Brazilian coast, high numbers of the
small brittle star Ophiactis savignyi usually live associ-
ated with the sponge Geodia corticostylifera (Demo-
spongiae, Geodidae), but not with other sympatric
sponge species. In order to check whether this associa-
tion was related only with the physical shelter provided
by the sponge body or was chemically mediated, the
crude organic extract of G. corticostylifera was added to
sponge mimics made of phytagel and spongin skeleton.
Control and treated mimics were simultaneously offered
to previously sponge-associated O. savignyi in both
static seawater and flow-through laboratory experi-
ments. Ophiuroids were allowed to move towards the
preferred mimic. The defensive properties of the sponge
extract against fish predation and fouling were also
evaluated. Chemotaxis assays showed that symbiotic
ophiuroids were able to chemically recognize its host
sponge, moving significantly more towards mimics
containing G. corticostylifera extract. Chemical deter-
rence assays showed that the natural concentration of
the extract of this sponge was also able to inhibit gen-
eralist fish predation on field experiments and the
attachment of the common mussel Perna perna in lab-
oratory assays. These results indicate that the crude
extract of G. corticostylifera plays multiple functions in
Communicated by P.W. Sammarco, Chauvin
E. E. G. Clavico Æ B. A. P. da Gama Æ D. Batista
R. C. Pereira (&)
Pós-Graduação em Biologia Marinha,
Universidade Federal Fluminense,
PO Box 100.644, 24001-970
Niterói, Rio de Janeiro, Brazil
E-mail: egbrecp@vm.uff.br
Tel.: 55 21 26292311
Fax: 55 21 26292292
G. Muricy Æ C. R. R. Ventura
Museu Nacional, Departamento de Invertebrados,
Universidade Federal do Rio de Janeiro,
20940-040 Rio de Janeiro, Brazil
the marine environment, presumably being responsible
for a closer association of this sponge with O. savignyi,
providing protection for this ophiuroid and inhibition of
epibionts on itself.
Introduction
Marine sponges are conspicuous members of the marine
benthos, occurring worldwide from polar and temperate
to tropical seas, from the intertidal to deep-sea envi-
ronments (Sarà and Vacelet 1973; Bergquist 1978;
Hooper and van Soest 2002). These benthic organisms
are particularly prominent, together with scleractinian
and alcyonarian coelenterates, in coral reef environ-
ments, which are commonly characterized by high levels
of herbivory and predation (e.g., Rützler 1978; Huston
1985; Hay 1991). In Caribbean coral reefs, sponges are
one of the most abundant and diverse groups of sessile
invertebrates, and they represent an important group for
ecological studies (Alcolado 1990; Diaz and Rützler
2001; Wulff 2001). As in the Caribbean, sponge species
are very abundant along the Brazilian coast, although
the majority of the species from the broad coastline of
this southwestern Atlantic region remain still unknown
(Hadju et al. 1999).
Sponges are the major source of unusual secondary
metabolites from marine organisms (Munro et al. 1999;
Faulkner 2002 and previous reviews from this author).
These compounds are known to exhibit a broad spectrum
of biological activities, such as antitumoral, antifungal,
antiviral and antibacterial (e.g., Green et al. 1990; Mur-
icy et al. 1993; Garson 1994; Munro et al. 1994; Almeida
et al. 1996). Despite this, only recently ecological exper-
iments have cast some light upon the true ecological roles
of secondary metabolites from marine sponges.
Some previous studies performed feeding assays
employing appropriate methodology in order to evalu-
ate the effectiveness of crude extracts or pure com-
pounds from sponges as defenses against predators
480
mainly in the tropical and temperate Pacific, Mediter-
ranean and Caribbean (Cimino et al. 1982; Thompson
et al. 1985; Pawlik et al. 1988; Herb et al. 1990; Rogers
and Paul 1991; Duffy and Paul 1992; McClintock et al.
1994; Pennings et al. 1994), and from a single species in
the Southwestern Atlantic (Epifanio et al. 1999). A wide
study accomplished by Pawlik et al. (1995) demon-
strated that the majority (69%) of the 71 species of
Caribbean sponges yielded crude extracts deterrent
against the reef fish Thalassoma bifasciatum (Bloch
1791), but also observed a considerable inter- and
intraspecific variability in deterrency. Although chemical
defense was considered an important strategy for most
Caribbean sponges (Pawlik et al. 1995), some common
species appear to use other tactics such as spicules both
in the Caribbean and Red Sea (Burns and Ilan 2003).
Although the most common ecological function
attributed to the secondary metabolites of sponges seems
to be predation deterrence, other interesting roles are also
known. For example, an allelopathic interaction was
verified in Guam, where the sponge Dysidea sp. over-
grows Cacospongia sp. after causing necrosis on it
(Thacker et al. 1998). Recently, a pioneer field study
provided evidence that the secondary metabolites of
sponges may really act as allomones by preventing over-
growth, or, on the other hand, as kairomones by
enhancing overgrowth by others (Engel and Pawlik 2000).
In addition, many studies have shown that secondary
metabolites extracted from a diversity of marine sponges
may inhibit fouling in the laboratory (e.g., Hirota et al.
1998; Tomono et al. 1998; Tsukamoto et al. 1998) or in
field assays (e.g., Bakus et al. 1990; Pereira et al. 2002),
thus suggesting that sponges remain devoid of epibionts
through the production of allelochemicals. However,
most sponges form symbiotic relationships with various
heterotrophic and autotrophic bacteria, protozoa, mic-
roalgae and invertebrates, and these interdependencies
develop to the point that for some individuals more of
the biomass is contributed by the symbionts than by the
sponge (Bertrand and Vacelet 1971).
Sponges harbour a rich associated fauna on their
surfaces, canals and inner tissues (e.g., Pearse 1932,
1950; Forester 1979; Koukouras et al. 1985; Klitgaard
1995; Neves and Omena 2003; Ribeiro et al. 2003;
Henkel and Pawlik 2005), that they are considered as
‘‘live hotels’’ (Pearse 1932) and one of the richest marine
benthic biotopes (Klitgaard 1995). The composition of
the sponge-associated fauna may be affected by different
characteristics of the sponges, such as size (Pearse 1932;
Frith 1976), morphology and anatomy (Koukouras
et al. 1985; Klitgaard 1995; Neves and Omena 2003), or
by the geographic distribution (Pearse 1950; Ribeiro
et al. 2003), depth (Peattie and Hoare 1981) and sea-
sonality (Cuartas and Excoffon 1993). Nonetheless, few
detailed surveys have been made on the production of
chemical cues by sponges that may attract or repel
particular animal species. Antibiotic substances from the
sponge Halichondria panicea (Pallas 1766) can be de-
tected by amphipods (Jakowska and Nigrelli 1960). The
associated amphipods, Caprella linearis (Linnaeus 1767)
and Jassa falcata (Montagu 1818) were attracted to H.
panicea in laboratory experiments (Peattie and Hoare
1981). In the presence of the same sponge species, there
was inhibition of the swimming ability of females of
another amphipod, Tritaeta gibbosa (Bate 1862) (Pea-
cock 1971). The amphipods Microdeutopus damnoniensis
(Bate 1856), M. anomalus (Rathke 1843) and Corophium
sextoni (Crawford 1937), as well as the crab Carcinus
maenas (Linnaeus, 1758), were attracted to H. panicea
and Hymeniacidon perleve (Montagu 1818) (Frith 1977).
These examples indicate that the host sponges may
produce chemical attractants that act as cues for the
associated fauna. Recently Henkel and Pawlik (2005)
also suggested that sponge-dwelling brittle stars must
have some mechanism for sponge habitat recognition.
Despite the large literature about ecological roles of
sponge chemicals in the tropical Pacific (e.g., Rogers and
Paul 1991; Becerro et al. 2003), Mediterranean (e.g.,
Uriz et al. 1992; Becerro et al. 2003), Bermuda
(e.g., McClintock et al. 1994) and the Caribbean sea
(e.g., Chanas and Pawlik 1995; Pawlik et al. 1995;
Puyana et al. 2003), these investigations are rare in the
South Atlantic region. Epifanio et al. (1999) demon-
strated that the compound variabilin, isolated from the
sponge Ircinia strobilina (Lamarck 1816) from the Ab-
rolhos archipelago, Brazil, deters fish predation, and
Pereira et al. (2002) demonstrated the antifouling
activity of crude extracts from Mycale microsigmatosa
(Arndt 1927), which acted selectively against barnacles
in field experiments. In addition, few experimental
studies have attempted to demonstrate the involvement
of sponge chemicals in multiple ecological roles.
The sponge Geodia corticostylifera Hajdu et al. 1992
(Demospongiae, Geodidae) possesses undescribed sec-
ondary metabolites exhibiting several pharmacological
properties, such as hemolytic, cytotoxic, neurotoxic
(Rangel et al. 2001), and antimicrobial activities (Mur-
icy et al. 1993). Some brittle star species live associated
with this sponge (Hajdu et al. 1992), in particular Oph-
iactis savignyi (Müller and Troschel 1842), which is
found inside the aquiferous canals of the sponge in
densities as high as four individuals per cm3 of sponge
tissue, or ca. 200 individuals per sponge (Clavico 2002).
We have hypothesized that O. savignyi is attracted to
G. corticostylifera by means of chemical cues, which may
also deter predators and epibionts. Thus, the aim of this
work was to evaluate the functional role of secondary
metabolites of G. corticostylifera as chemical cues to
associated brittle stars, as defense against generalist fish,
and as antifoulant against mussels.
Materials and methods
Organisms
The sponge G. corticostylifera is known to occur at
depths of 3–39 m in the upwelling coast of Arraial do

Cabo (southeastern Brazil), but may be found in all
southeastern Brazilian coast and also in Venezuela.
G. corticostylifera is orange externally and beige inter-
nally; it has a massive-globose, cerebriform shape, with
deep grooves at the surface, and a firm consistency
(Hadju et al. 1992).
The ophiuroid O. savignyi is widely distributed both
in tropical and subtropical regions, throughout the
Indo-Pacific, eastern Pacific and on both sides of the
Atlantic. In the Western Atlantic it is found in South
Carolina, Bermudas, the Caribbean, Gulf of Mexico and
Brazil. It usually inhabits shallow waters, but may also
be found down to 518-m depth (Hendler et al. 1995). At
Arraial do Cabo, this ophiuroid is the major component
of the endobiotic macrofauna of the sponge G. cortico-
stylifera (Hajdu et al. 1992; Clavico 2002).
Collection sites
Arraial do Cabo (Rio de Janeiro state, Brazil—2344’S–
4200’W) is a tropical peninsula projecting from the
Brazilian southeastern coast. This region is character-
ized by a coastal upwelling, created by the prevailing
northeastern and eastern winds (Valentin 1984; Gonz-
alez et al. 1992; Carbonel and Valentin 1999). Specimens
of G. corticostylifera and associated O. savignyi were
manually collected through SCUBA diving at Forno
beach and Saco dos Cardeiros, from 5 to 8-m depth.
Both collection sites are not directly affected by the
upwelling phenomenon. Specimens were enmeshed in
polyethylene bags during collection in order to prevent
the loss of associated fauna, which were separated from
the sponges in the laboratory.
For the antifouling assay, juvenile mussels (Perna
perna Linnaeus 1758) were collected during low tide
from the rocky coastal area of Itaipu (Niterói city, Rio
de Janeiro, Brazil—2354’30’’S and e 4302’00’’W) and
kept in a 230 l recirculating laboratory aquarium
(equipped with biological and activated carbon filtering,
and protein skimming) at constant temperature (20C),
salinity (ca. 35) and aeration for 12 h. Individuals were
then disaggregated by carefully cutting the byssus
threads, and divided into size groups according to total
shell length, ranging from 0.8 to 4.1 cm, in a plastic tray
with seawater.
Ophiuroid density estimation
A number of sympatric sponge species was sampled
(n=1–12, depending on the natural abundance of each
species), enmeshed in polyethylene bags and the density
of associated O. savigny (ind. 100 ml 1) was estimated
in the laboratory. Sponge volume was measured
according to Rützler (1978), this is, by water displace-
ment in a graduated cylinder.
481
Extraction procedures
Soon after collection, samples of G. corticostylifera were
rinsed thoroughly in seawater to remove any associated
animals, which may cause the undesired interference of
substances other than secondary metabolites produced
by the sponge or their endobiotic bacteria. Sponge tis-
sues were cut in small pieces (ca. 1 cm) and successively
extracted with organic solvents. The first extraction was
done by homogenizing sponge pieces in methanol
(MeOH) prior to exposition to ultrasonic waves for 15
min to increase the effectiveness of extraction. A second
extraction was made with a mixture of MeOH and di-
chloromethane (DCM) at the proportion of 1:1. The
extract was filtered and all solvents were eliminated in a
rotatory evaporator under reduced pressure.
Chemotaxis
Control and treatment sponge mimics were prepared
using pieces of commercial marine sponges (dry spongin
skeleton) to which an extract-containing gel (treatments)
or gel containing only solvent (controls) were added as
follows. The natural volumetric concentration of the
extract was obtained by using previously prepared crude
organic extract corresponding to 20 ml of G. cortico-
stylifera to prepare 20 ml of gel. A total of 1.01 g of
phytagel (Sigma Chemical Co.) was dissolved in 20 ml
of deionized water and vigorously stirred prior to heat-
ing to the boiling point. After cooling down to ca. 60C
the extract aliquot was added (dissolved in DCM) and
the mixture was then poured into one spongin piece
(treatment). Control mimics were made in the same
manner, but adding DCM only to control for solvent
effects. Phytagel was used so as to allow the controlled
diffusion of organic extract from the sponge mimic to
seawater in a manner similar to that supposed to occur
under natural circumstances (Henrikson and Pawlik
1995; da Gama et al. 2002).
Two kinds of experiments were carried out to verify
the effectiveness of chemicals from G. corticostylifera as
a taxis signal to O. savignyi, static and flow-through
experiments.
Static seawater experiment
In the first experimental trials, control and treatment
mimics were placed in a rectangular plastic container
together with one specimen of O. savignyi. All recipients
(perforated plastic containers; n=30) were placed in a 2-
m diameter fiberglass pool with a water depth of 1.0 m,
without water flow. One brittle star was placed equi-
distantly from control and treated mimics in each con-
tainer (Fig. 1a). All ophiuroids that moved toward
control or treatment and remained there after 12 h were
counted as having been attracted to one of these options.
482
Flow-through experiment
As static seawater experiments are unrealistic in nature, a
second set of experiments was performed employing a
seawater flow-through design in order to simulate more
realistic conditions of flow and dilution of possible
chemical cues. A similar assay was performed, but con-
trol and treatment mimics were arranged in opposite
angles of a rectangular plastic tray (2 l) with an input
and output of seawater. Control and treated mimics were
positioned at both sides of the water input, so that the
water could flow to the ophiuroid (Fig. 1b). At the end of
each assay (ca. 10 min), ophiuroids sheltering in control
or treatment mimics were counted (n=39). Brittle stars
that did not move to one of the choices or evaded the
containers were not counted in both experiments.
Chemical defense against fish predation
Field experiments and artificial food preparation were
performed according to Pawlik et al. (1995). An extract
equivalent to 60 ml of G. corticostylifera fresh tissue
was added to 0.3 g of alginic acid, tuna fish and water to
yield a final volume of 60 ml, in order to reproduce the
natural concentration of substances found in G. corti-
costylifera. The mixture was vigorously stirred and then
poured on a rectangular mold for pellet preparation
Fig. 1 a Scheme of
experimental unit used in the
a
static seawater experiment.
b Scheme of one experimental
unit of the flow through
chemotaxis experiment
evidencing the seawater flow
from the mimics to the brittle
star
water
input
(about 1.0·0.5·5.0 cm). Control pellets were made in
the same way, but without addition of extract (solvent
only). Field assays were performed at Forno beach,
Arraial do Cabo, at 3–6-m depth. Ropes containing a
pair of pellets (one control and one treatment, n=20)
were attached to the sea bottom, and exposed to gen-
eralist fish during a period of time sufficient to obtain a
measurable consumption (ca. 3 h). After this period the
remaining pellet was measured with a ruler. Weight of
pellets was not used since controls and treatments con-
taining extracts may differ in their absortion of dissolved
ions from seawater. The main generalist fish in Arraial
do Cabo, which might have been able to consume the
pellets, are Abudefduf saxatilis (Linaeus 1758) (Poma-
centridae), Haemulon aurolineatum (Cuvier 1830) (Hae-
mulidae), H. steindacneri (Jordan and Gilbert 1882)
(Haemulidae), Halichoeres poeyi (Steindachner 1867)
(Labridae), Chaetodon striatus (Linnaeus 1758) (Chae-
todontidae) and Kyphosus sectatrix (Linnaeus 1766)
(Kyphosidae) (Floeter et al. 2003).
Chemical defense against fouling
After collection of the mussels, individuals exhibiting
substrate-exploring behavior (actively exposing their
foot and crawling) were selected for experiments. Anti-
fouling activity was measured by the method described
Treatment Control
N=30
water
output

by da Gama et al. (2003), a modification of the proce-
dure first used by Ina et al. (1989) and Goto et al.
(1992). This laboratory experiment has been shown to
correlate well with field results (da Gama et al. 2003).
Water-resistant filter paper was cut into 9-cm diameter
circles and soaked in solvent (control filter). Another 9-
cm diameter set of filter papers (treatment filters) was cut
in a chess board pattern (1-cm squares) and soaked in a
natural concentration of G. corticostylifera crude ex-
tract. Natural concentration was determined as the ex-
tract equivalent to the dry weight (DW) of sponge tissue
= filter paper, this is, 0.0835 g of extract, taken from
0.3094 g of sponge DW were embedded in 0.3094 g of
filter paper. Sponge volume was not used to reproduce
natural concentrations in this case, since filter paper is a
two-dimensional object. All filter paper circles were al-
lowed to air dry. Entire filter circles were then placed in
the bottom of sterile polystyrene Petri dishes, over which
treated chess board filters were placed. Dishes were filled
with 80 ml of seawater and three mussel specimens (2.0–
3.0-cm length) were added. Thus, mussels would have
the same area of treated (superior, squared) and control
(inferior, entire) filter paper to which they attach. Ten
replicates of each treatment (blank control and G. cor-
ticostylifera extract) were used. Experimental dishes
were kept in total darkness, as mussels have been shown
to produce more byssal threads when held in the dark
(Davis and Moreno 1995). Experiments were allowed to
run for 12 h. Mussel activities were recorded immedi-
ately after the start of the experiment, after 2 h, and
then after 12 h. The activities recorded were substra-
tum-exploring behavior, gamete release (as an indicator
of possible stress or positive cues) and the number of
byssal threads attached to each substratum (control or
treated filter paper, shell of another mussel or border of
Petri dish). After the 12-h period, all records of attach-
ment were checked, and the mussels were placed in mesh
bags tagged according to treatment and suspended in a
sea aquarium for 24 h to check for possible mortality
due to exposure to the test substances.
Statistical analyses
The statistical analyses were applied according to the
experimental design of each assay. The data from anti-
fouling assays were analyzed as the number of attached
byssal threads, and field predation data were analyzed as
the amount (in mm) of the pellet consumed (not as
percentages as shown in graphs). Yates’ corrected chi-
square (v2) was used to analyze chemotaxis results, un-
der the null hypothesis of an equal proportion of brittle
stars moving to each treatment. Antifouling and pre-
dation results were analyzed by the Wilcoxon signed
ranks test, a nonparametric equivalent to the t test for
dependent samples, as variables did not follow the
assumption of normal distribution (Zar 1999). Differ-
ences among treatments were considered significant only
when p<0.05 (a=5%).
483
Results
Ophiuroid density
A total of 27 specimens of six sponge species were
sampled: Amphimedon viridis (Duchassaing and Mi-
chelotti 1864); Aplysina fulva (Pallas 1766), Dysidea ro-
busta (Vilanova and Muricy 2001), G. corticostylifera
(Hajdu et al. 1992) and Paraleucilla magna (Klautau
et al. 2004). The density of O. savigny ranged from 0 to
385 ind. 100 ml 1 of sponge (Table 1). The highest
densities were found in G. corticostylifera (mean=76
ind. 100 ml 1) and the lowest density in A. fulva
(mean=1 ind. 100 ml 1). As an unequal number of
individuals were collected from each species, we did not
perform statistical analyses with these data.
Chemotaxis
O. savignyi showed a greater tendency to move towards
G. corticostylifera than towards the control in static
seawater assays, but this trend was not statistically sig-
nificant (v2, p<0.37; Fig. 2a). In contrast, significantly
higher numbers of O. savignyi moved towards sponge
mimics containing G. corticostylifera crude extract in
flowing seawater trials (v2, p<0.03). This result indicates
the presence of substances capable of inducing positive
taxis behavior on the associated ophiuroid O. savignyi in
G. corticostylifera.
Chemical defense against fish predation
The assays for chemical defensive role against predation
by generalist fish showed that artificial food containing
the crude extract of G. corticostylifera (treatments) was
significantly less consumed than the control (Wilcoxon,
p<0.001; Fig. 2b). This result demonstrated that the
crude extract of this sponge species is active as a defense
against a diverse array of generalist fish, including
Abudefduf saxatilis, Haemulon aurolineatum, H. stein-
dacneri, and Kyphosus sectatrix, the most common spe-
cies in the area studied.
Chemical defense against fouling
The crude extract of G. corticostylifera significantly
inhibited byssal thread attachment of juvenile P. perna
relative to the control (Wilcoxon, p<0.01), showing a
very strong antifouling activity (Fig. 2c). After the 24 h
recovery period, a high mortality rate (66%) of the
mussels previously exposed to the sponge extract was
recorded, whereas none from the controls died (Fig. 2d).
Therefore, the mechanism of attachment inhibition in
the antifouling assays is probably related to the toxicity
of the crude extract of G. corticostylifera to the mussels.
484

Table 1 Number (N), dominance (%) and richness of ophiuroid species in each sponge species

Ophiuroids Ophiactis Ophiactis Ophiothrix Amphipholis

savignyi lymani angulata squamata

Sponges N Percentage N Percentage N Percentage N Percentage N per species Species richness

Aplysina fulva 24 100 – – – – – – 24 1

Dysidea robusta 15 75 3 15 1 5 1 5 20 4
Arenosclera brasiliensis 7 70 2 20 1 10 – – 10 3
Amphimedon viridis 249 98 – – 5 2 254 2
Paraleucilla magna 6 100 – – – – – 6 1
Geodia corticostylifera 1,477 99.8 2 0.1 2 0.1 – – 1,479 3

Fig. 2 a Movement of the brittle star O. savignyi towards control or treatment sponge mimic in static seawater and flow through
chemotaxis assays. b Percentual mass eaten of the control and treatment pellets after the field assay by generalist fish. c. Attachment of the
mussel P. perna (mean number of byssal threads) in control and treatment after 12 h. d. Percent survivorship of the mussel P. perna 24 h
after the end of the experiments. Error bars are standard deviations. N S = not significant

G. corticostylifera, but not in other sympatric sponges. A

Discussion
The brittle star O. savignyi is commonly found in a
number of habitats (Hendler et al. 1995) including living
organisms, such as algae (Mladenov and Emson 1988),
polychaete mats (Nalesso et al. 1995; Britayev et al.
1999) and sponges (Duarte and Nalesso 1996; Carrera-
Parra and Vargas-Hernandez 1997). High numbers of
individuals of this species were found associated to
similar pattern was found for Ophiothrix fragilis, which
were found abundantly in some sponge species, and
clearing experiments suggested that these brittle stars
can select their habitat after settlement (Turon et al.
2000). Our results showed that G. corticostylifera pro-
duces secondary metabolites, which attract the ophiu-
roid O. savignyi. In nature, chemical cues probably
trigger the association between G. corticostylifera and

O. savignyi, corroborating previous evidences that the
behavior of ophiuroids is largely intermediated by che-
moreception (e.g., Sloan and Campbell 1982).
Probably, O. savignyi seeks the association with
G. corticostylifera for reasons related mainly to feeding,
defense, or both. Organisms associated to sponges may
use the constant water flow through the aquiferous
system of sponges to obtain particulate food (Westinga
and Hoetjes 1981). Klitgaard (1995) suggested that the
large amount of organisms associated to specimens of
Geodia from Faroe Islands is related to the abundance of
detritus accumulated on the sponge surface. Based on its
stomach contents, O. savignyi is considered a detritivo-
rous, whose diet largely consists of detritus, sand grains,
bryozoans, foraminiferans and the remains of small
gastropods (Boffi 1972). According to Emson and
Mladenov (1992), this species may also be a suspension
feeder that uses its arms to catch current-borne particles
and selectively transfer them to the mouth. Harrold and
Reed (1985) suggested that the activity of some echino-
derms like sea urchins and sea stars may be influenced
by nutritional conditions, and this may prove important
in brittle stars, too. Thus, G. corticostylifera could pro-
vide food to O. savignyi by the constant flow of water
containing plankton or organic particles too large to be
used by the sponge. An alternative hypothesis is that
O. savignyi feeds directly on G. corticostylifera tissues,
but there are no convincing evidences supporting it.
Since the crude extract from G. corticostylifera con-
sistently deterred generalist fish in the field, it is rea-
sonable to infer that the sponge may also provide
protection against predation on O. savignyi. Randall
(1967) reported 33 species of coral reef fishes in the
Caribbean that preyed upon ophiuroids. Furthermore,
factors like their cryptic and nocturnal behaviors indi-
cate the incessant predation pressure in these organisms
(Birkeland 1989). These aspects may corroborate the
idea that the shelter provided by G. cortycostilifera may
protect O. savigny against predation. On the other hand,
Henkel and Pawlik (2005) suggested that sponge-asso-
ciated brittle stars do not seem to gain an associational
chemical defense against generalist fish predators.
The shelter provided by the sponges may be one of
the main reasons why these animals possess a rich-
associated fauna. The predation-deterrent activity of the
secondary metabolites of sponge can be an important
factor to determine the composition of sponge-associ-
ated fauna. In general, defensive compounds against
fishes and sea urchins do not affect small and relatively
sedentary herbivores, such as amphipods, polychaetes
and crabs (Hay and Duffy 1990). Mesograzer herbivores
subjected to high predation rates may preferentially feed
or live on chemically-defended plants to minimize their
susceptibility to natural enemies (Hay and Steinberg
1992). It is therefore likely that the ophiuroid O. savignyi
is immune to the antifouling and antipredation com-
pounds of G. corticostylifera, and that it inhabits this
chemically defended sponge to defend itself against
predation. It remains, however, unclear whether
485
G. corticostylifera takes any benefits or disadvantages
from the association with O. savignyi. The ophiuroid
may eat large particles that could otherwise block the
inhalant canals and reduce water flow in the sponge
aquiferous system; on the other hand the body of the
ophiuroid itself is likely to reduce water flow in the
aquiferous system. On average, the net result for the
sponge might be negative, but this remains to be
experimentally demonstrated.
The ‘mussel test’ performed for antifouling activity is
a reliable time and cost-saving screening method, whose
results are generally reproduced by field assays (da
Gama et al. 2003). It is thus reasonable to assume that
the extract of G. corticostylifera could inhibit the fixation
of epibionts in situ. This is corroborated by field
observations that the surface of G. corticostylifera is
often (but not always) clean of macroscopic epibionts
(Hajdu et al. 1992; G. Muricy, personal observation).
Many soft-bodied marine invertebrates have evolved
chemical defense strategies to suppress epibiosis,
including secondary metabolites produced by sponges
(e.g., Hirota et al. 1998; Pereira et al. 2002). The nega-
tive effects of fouling on marine host organisms (mostly
macroalgae) have been extensively documented, and
include a decrease of light and nutrient availability, a
loss of flexibility resulting in increased brittleness,
mechanical damage of host surfaces, changes of surface
pH and changes in drag that can cause breakage during
storms. For example, host plants can be fouled by epi-
bionts that are preferred by herbivores, thus suffering
not only the direct negative effects of being fouled (Davis
et al. 1989; Wahl 1989; Williams and Seed 1992), but
also experiencing increased rates of grazing (Wahl and
Hay 1995; Karez et al. 2000; Pereira et al. 2002). All
these processes could select for characteristics that
minimize colonization by fouling organisms, for exam-
ple, the production of chemical defenses against epibi-
osis.
Our results showed that the crude extract of G. cor-
ticostylifera has multiple ecological roles. It seems to act
as a chemical cue for shelter to O. savignyi, as a chemical
defense against generalist fish, and as an antifouling
agent against the mussel P. perna in laboratory assays. It
also caused high mortality in mussels after the anti-
fouling experiments, corroborating previous observa-
tions that extracts from G. corticostylifera are highly
toxic (Rangel et al. 2001). These extracts also have
antifungal and antibacterial properties, but with a rela-
tively weak and selective activity directed mainly against
Staphylococcus aureus (Rosenbach 1884; Muricy et al.
1993).
Due to the high molecular complexity and relatively
high concentrations of secondary metabolites found in
marine organisms, it has been suggested that defenses
are costly and thus the benefits of chemical defenses
should increase fitness more than the costs reduce fitness
(Cronin 2001). If G. corticostylifera could deter an array
of adverse factors such as predation, epibiosis, and
infections by pathogenic microorganisms with a single
486
compound, it would gain an advantage in fitness over
another species that needs to produce separate com-
pounds for each function. Marine secondary metabolites
with multiple functions have already been demonstrated
to exist (Schmitt et al. 1995; da Gama et al. 2003;
Pereira et al. 2003). A single secondary metabolite can
deter feeding by herbivores (Pereira et al. 2003) and in-
hibit fouling settlement in the field (da Gama et al.
2003). In G. corticostylifera, however, the nature and
number of compounds responsible for the observed
activities still remain to be demonstrated, since no de-
tailed chemical study has yet been done on this species.
Acknowledgements The authors wish to thank the staff of the In-
stituto de Estudos do Mar Alte. Paulo Moreira (IEAPM—Brazil-
ian Navy), in special CEL Ferreira for support during field work.
R.C. Pereira, C.R.R.Ventura and G.Muricy thank CNPq for their
Research Productivity fellowships, and B.A.P.da Gama thanks
CAPES for his ProDoc fellowship. G.Muricy also thanks FAPERJ
for a grant ‘‘Cientistas do Nosso Estado.’’ Anonymous referees
greatly improved the quality of this paper. The experiments de-
scribed here comply with the Brazilian environmental protection
laws.
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