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Sponges as substrata and early life history of thetubulariid Zyzzyzus warreni (Cnidaria: Hydrozoa) inthe São Sebastião Channel, BrazilCarlos José Alexandre De Campos a , Alvaro Esteves Migotto b c , Ulisses Pinheiro d &Antonio Carlos Marques ca Centre for Environment, Fisheries & Aquaculture Science (Cefas), Weymouth, UKb Centro de Biologia Marinha, Universidade de São Paulo, São Sebastião, Brazilc Departamento de Zoologia, Instituto de Biociências, Universidade de São Paulo, SãoPaulo, Brazild Departamento de Zoologia, Centro de Ciências Biológicas, Universidade Federal dePernambuco, Recife, Brazil

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To cite this article: Carlos José Alexandre De Campos, Alvaro Esteves Migotto, Ulisses Pinheiro & Antonio Carlos Marques(2012): Sponges as substrata and early life history of the tubulariid Zyzzyzus warreni (Cnidaria: Hydrozoa) in the SãoSebastião Channel, Brazil, Marine Biology Research, 8:7, 573-583

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ORIGINAL ARTICLE

Sponges as substrata and early life history of the tubulariid Zyzzyzus

warreni (Cnidaria: Hydrozoa) in the Sao Sebastiao Channel, Brazil

CARLOS JOSE ALEXANDRE DE CAMPOS1*, ALVARO ESTEVES MIGOTTO2,3,

ULISSES PINHEIRO4 & ANTONIO CARLOS MARQUES3

1Centre for Environment, Fisheries & Aquaculture Science (Cefas), Weymouth, UK, 2Centro de Biologia Marinha,

Universidade de Sao Paulo, Sao Sebastiao, Brazil, 3Departamento de Zoologia, Instituto de Biociencias, Universidade de

Sao Paulo, Sao Paulo, Brazil, and 4Departamento de Zoologia, Centro de Ciencias Biologicas, Universidade Federal de

Pernambuco, Recife, Brazil

AbstractThe hydroid Zyzzyzus warreni is usually found in shallow coastal waters forming aggregations of solitary polyps embeddedin demosponges. Early life history transformations and settlement responses by the actinulae of this hydroid were studied inthe laboratory using 13 species of sponges and 2 species of algae collected in the Sao Sebastiao Channel (Brazil) assubstrata. The absence of oral tentacles and mouth in the actinulae and early events of metamorphosis suggest that theselarvae are unable to spend long periods in the plankton and attach quickly near conspecifics when a preferred substratum isencountered. The time required for settlement and the number of elicited settlings indicated four settlement responses: (a)frequent and short-time settlement, in actinulae exposed to Halichondria cebimarensis, Mycale angulosa, M. aff. americana,M. laxissima (skeleton) and Tedania ignis; (b) moderate and delayed settlement, in actinulae exposed to Aplysina caissara,A. fulva, Haliclona melana and M. microsigmatosa; (c) no settlement, in actinulae exposed to Suberites aurantiacus and to thealgae Hypnea musciformis and Sargassum cymosum; and (d) lethal response, in actinulae exposed to Amphimedon viridis,Aplysilla rosea, Dragmacidon reticulatum and M. laxissima. These responses indicate a considerable degree of speciesdiscrimination by the actinulae and are consistent with substrata used by the hydroid in the natural environment.

Key words: Porifera, Hydrozoa, Zyzzyzus warreni, settlement, life history, feeding behaviour

Introduction

Most hydroids have complex metagenetic life his-

tories which include the benthic polyp and the

pelagic medusa. In these animals, the metamorpho-

sis to the developed polyp involves a series of

morpho-physiological transformations, which may

occur by means of different strategies, including the

development of different larval forms (see Berking

1998; Muller & Leitz 2002). The life history of

tubulariid hydroids includes the development of a

lecithotrophic larva called actinula, which contains

tentacles formed inside the reproductive structure �the gonophore � just before release into the water

column (Berking 1998).

Aspects of the life history of tubulariids have been

described in the literature (e.g. Pyefinch & Downing

1949; Berrill 1952). However, very little is known

about the role of the actinulae on dispersal, selection

of substrata and demography of tubulariids.

Sponges are used as substrata by many species of

hydroids, which may thrive on or inside the sponges

(Puce et al. 2005). The potential benefits to both

groups of animals presumed to exist in this associa-

tion include trophic and reproductive aspects, such

as refuge from predators and protection against

water movement (Bacescu 1971; Klitgaard 1995;

Duarte & Nalesso 1996; Ribeiro et al. 2003). For

instance, anthoathecate tubulariids (e.g. species of

*Correspondence: Carlos Jose Alexandre de Campos, Centre for Environment, Fisheries & Aquaculture Science (Cefas), Weymouth

Laboratory, Barrack Road, The Nothe, Weymouth, Dorset DT4 8UB, UK. E-mail: carlos.campos@cefas.co.uk

Published in collaboration with the University of Bergen and the Institute of Marine Research, Norway, and the Marine Biological Laboratory,

University of Copenhagen, Denmark

Marine Biology Research, 2012; 8: 573�583

(Accepted 4 October 2011; Published online 25 May 2012)

ISSN 1745-1000 print/ISSN 1745-1019 online # 2012 Taylor & Francis

http://dx.doi.org/10.1080/17451000.2011.638641

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Hybocodon and Zyzzyzus) are commonly found on

demosponges (see Warren 1906; Hirohito 1988;

Petersen 1990).

Zyzzyzus warreni Calder, 1988 has been observed

in sheltered shallow coastal waters in Bermuda

(Calder 1993), on rocky shores in southeastern

Brazil (Migotto & Silveira 1987; Migotto 1996)

and French Lesser Antilles (Galea 2008), mangrove

roots in northeastern Brazil (Calder & Mayal 1998)

and Belize (Calder 1991), bays in South Africa and

Mozambique (Millard 1975) and the Netherlands

Antilles (de Kruijf 1977), and in protected areas in

Japan (Hirohito 1988), Puerto Rico (Wedler &

Larson 1986) and Colombia (Bandel & Wedler

1987), forming aggregations of polyps on a number

of different species of sponges (Warren 1906; Mi-

gotto & Silveira, 1987; Calder 1988).

The objective of this study was to investigate the

degree of substrate selection by the actinulae of

Z. warreni and describe the early life history of this

hydroid. This investigation was undertaken with the

eventual intention of providing new evidence to-

wards clarification of the taxonomic position of this

species, which has been much debated (Petersen

1979, 1990; Marques & Migotto 2001; Campos

et al. 2007).

Materials and methods

Life history studies and settlement experiments were

undertaken between February and April 2002, when

many polyps of Zyzzyzus warreni were releasing

actinulae. Actinulae and polyps were reared in Petri

dishes containing filtered sea water (FSW) and fed

with Artemia nauplii. Polyps were measured under a

Zeiss Axiophot microscope equipped with Zeiss KS

300, after being anaesthetized in a 1:1 solution of

7.5% MgCl2 and seawater and preserved in 4%

formaldehyde. The cnidome (nematocyst composi-

tion and measurements) was studied in actinulae

and juvenile polyps (3, 12 and 18 h old). Only

capsules of undischarged nematocysts were mea-

sured. The nematocyst nomenclature adopted is that

of Mariscal (1974) and Millard (1975).

Samples of sponges, polyps and algae were

hand-picked from three sites in the Sao Sebastiao

Channel (southeast Brazil). The sponges Mycale

angulosa (Duchassaing & Michelotti, 1864), Mycale

microsigmatosa Arndt, 1927, and Tedania ignis

(Duchassaing & Michelotti, 1864) containing polyps

were sampled from an iron frame deployed at depth

of 3 m in the shallow bay of Praia do Segredo

(23849?42??S; 45825?16??W). In addition, 10 other

species of sponges were collected from an adjacent

rocky shore at Ponta do Jaroba (23849?68??S;

45825?28??W): Amphimedon viridis Duchassaing &

Michelotti, 1864, Aplysina caissara Pinheiro &

Hajdu, 2001, Aplysina fulva Pallas, 1766, Aplysilla

rosea (Barrois, 1876), Dragmacidon reticulatum

(Ridley & Dendy, 1886), Haliclona melana (Muricy

& Ribeiro, 1999), Mycale aff. americana (Van Soest,

1984), Mycale laxissima (Duchassaing & Michelotti,

1864), Halichondria cebimarensis Carvalho & Hajdu,

2001, and Suberites aurantiacus (Duchassaing &

Michelotti, 1864) (Custodio & Hajdu 2011). The

red alga Hypnea musciformis (Wulfen) J. V. Lamour-

oux and the brown alga Sargassum cymosum C.

Agardh were sampled from an adjacent sandy beach,

Praia de Guarapocaia (Ilhabela) (23844?80??S;

45820?94??W). Samples were transported to the

laboratory in plastic containers (1500 ml) containing

seawater. In the laboratory, samples were transferred

to large plastic trays receiving continuous trickle flow

of FSW. Actinulae released from adult polyps were

collected from the water column on a daily basis

using a pipette and maintained in glass bowls

containing FSW (salinity�34 psu) kept under

constant room temperature (258C) with a 12 h

light/12 h dark photoperiod.

Settlement experiments were performed in 24-well

Cell Culture Clusters (Corning). Each well contain-

ing 3 ml of FSW received:0.25 cm3 of sponge or

algae as substrata and one actinula. Two sets of

experiments were conducted. In the first, all species

of sponges and algae were used. Each experiment

consisted of 15 actinulae per substratum and a

further 15 actinulae without substratum as control.

Actinulae exposed to M. laxissima died immediately

(see Results), probably due to the deleterious effect

of mucus released from the sponge (see Hajdu &

Rutzler 1998). To confirm this, fragments of

M. laxissima were rinsed with FSW and the clean

skeleton of the sponge was used in a further

experiment.

Six species of sponges (A. fulva, A. viridis,

H. melana, M. aff. americana, M. angulosa and

T. ignis) were selected for a second experiment.

This consisted of 45 actinulae per substratum and

15 actinulae without substratum as control. The

number of settled actinulae was recorded every 20

min during the first experiment (total duration�17

h) and every 10 min during the second experiment

(total duration�08:20 h). Seawater in culture cells

was replaced hourly. The emergence of a protrusion

in the aboral pole of the actinulae was the criterion

for considering a successful settlement. This juve-

nile-specific structure has been described in

Acharadria spp. (see Pyefinch & Downing 1949;

Berrill 1952).

The differences in the number of settled actinulae

between substrata in the second experiment were

analysed by one-way ANOVA, followed by Tukey’s

574 C. J. A. de Campos et al.

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family error test (a�0.05) for post-hoc comparisons

of means. Confidence intervals were determined for

pairwise differences between means. The first ana-

lysis was performed for the total number of settled

actinulae during the experiment and the second

analysis for the total number of settled actinulae at

60 min of settlement.

Representatives of sampled sponges were sub-

mitted to the Porifera collection of the Museu

Nacional, Universidade Federal do Rio de Janeiro

(MNRJ), Rio de Janeiro: MNRJ 6749 and 6758: A.

fulva; MNRJ 6750: M. microsigmatosa; MNRJ 6752

and 6753: H. melana; MNRJ 6754: S. aurantiacus;

MNRJ 6756: D. reticulatum; MNRJ 6757: A. rosea;

MNRJ 6759: A. viridis; MNRJ 6760: A. caissara;

MNRJ 6761: T. ignis. Representatives of collected

polyps were submitted to the Cnidaria collection of

the Museu de Zoologia da Universidade de Sao

Paulo: MZUSP 1983.

Results

Morphology and behaviour of actinulae

The morphology of polyps of Zyzzyzus warreni was

described previously (Campos et al. 2007). Fully

developed female gonophores contained between

one and four actinulae (Figure 1a). Actinulae were

released from the gonophore in a series of 2�19

Figure 1. Life history stages of Zyzzyzus warreni. Actinulae (a, b, c); newly metamorphosed polyps with 2 h (d), 10 h (e), and 30 h (f) on

the sponge Mycale angulosa; adult polyp with two developing male gonophores (g); female gonophore with developing actinulae (h). Scale

bars �250 mm.

Selection of substrata by Zyzzyus warreni 575

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(n�25) pulsations per minute and per event.

A small population of 25 adult polyps released 70

actinulae during a single night. Just before release

into the water column, actinulae projected their

aboral poles towards the circular apical orifice of

the gonophore.

At this stage, each actinula consisted of a cone-

shaped body 316�474 mm long and 330�587 mm

wide at the aboral pole, with one whorl of 6�16

aboral, concentrically disposed filiform tentacles,

764�1432 mm long and 27�97 mm wide at their

bases (Figure 1b,c). Tentacles were flattened basally

and rounded distally in cross-section. The distance

between adjacent tentacular bases was equivalent to

the diameter of tentacles (Figure 1B). Clusters of

four different types of nematocysts (small stenoteles,

heterotrichous microbasic euryteles, homotrichous

microbasic euryteles and desmonemes) were ob-

served along the tentacles and on the terminal

swollen tip. None of the:104 actinulae had devel-

oped a mouth at this stage.

Actinulae did not show active swimming ability

inside culture cells. However, their tentacles exhib-

ited slow movements around the edges of these

compartments with no permanent attachment ob-

served in any of the experiments (see below). Once

in contact with the preferred biogenic substrata,

most tentacles became orientated towards the sub-

stratum. This behaviour was not observed in acti-

nulae exposed to lethal sponges (see below).

Actinulae remained viable for at least 3 days if not

in contact with biogenic substrata.

Settlement

Actinulae of Zyzzyzus warreni exhibited different

settlement responses according to the species used

as substrata. No settlement or death of actinulae was

recorded in any of the controls.

There were no settlements on the algae Hypnea

musciformis and Sargassum cymosum. In the first

experiment, 8 of the 13 species of sponges elicited

settlement. There were no settlements on the sponge

Suberites aurantiacus, and the actinulae died in

contact with four species of sponges. Of these,

Dragmacidon reticulatum and Amphimedon viridis

were the most lethal, i.e. 100% of actinulae died

during the first 20 min of exposure. All actinulae

died within 2 h in the experiments with Aplysilla rosea

and Mycale laxissima. In all cases of contact with

lethal sponges, body maceration and tentacle resorp-

tion were observed during the first minutes of

exposure, leading to death of actinulae. The highest

rates of settlement were observed in Halichondria

cebimarensis (100% of actinulae settled within 40 min

in the first experiment).

In the second experiment, the time required for

settlement and metamorphosis ranged between 10

min (in actinulae exposed to Mycale angulosa and

Tedania ignis) and 6 h (with the exception of a single

outlier settlement at:17 h on Aplysina caissara).

Actinulae used in controls remained alive in all

experiments and during the whole duration of the

experiment (17 h).

The sponges Mycale aff. americana, M. laxissima

(skeleton), and T. ignis elicited settlement and

metamorphosis in more than 50% of the actinulae

within the first 60 min (Figure 2). High settlement

rates were observed in each of these sponges over the

following 4 h, on which 100% (M. aff. americana),

93% (M. laxissima) and 80% (T. ignis) of the

actinulae settled.

The sponges Aplysina caissara, A. fulva, Haliclona

melana, M. angulosa and M. microsigmatosa elicited a

lower number of successful settlements than those

observed in the first group of sponges (Figure 2).

None of these sponges elicited more than 30% of

successful settlements during the first hour of the

experiment. The ratio of settled actinulae

was �50% in the tests with A. caissara, A. fulva

and M. microsigmatosa (maximum after 4 h). Settle-

ment was B50% in the tests with H. melana (max-

imum of 13% at 2:40 h) and M. angulosa (maximum

of 47% at 3 h) (Figure 2).

The second experiment corroborated the inter-

species differences in settlement rates observed

during the first experiment, with the exception of

M. angulosa (Figure 3), on which settlement was

quicker (from 47% actinulae settled at 3 h to 98%

actinulae settled within the first hour). ANOVA

revealed statistically significant (F�93.28;

p�0.000) differences between the total number of

actinulae settled. The Tukey’s test suggested the

following three groups of sponges for 99% con-

fidence intervals: M. aff. americana, T. ignis and

M. angulosa; A. fulva; H. melana and A. viridis.

When considering 60 min as the reference settle-

ment time, T. ignis elicited a higher number of

successful settlements in the second experiment

(from 67 to 84%), whereas M. aff. americana

elicited a lower number of successful settlements

in the second experiment (from 60 to 40%). T. ignis

elicited higher numbers of successful settlements in

the second experiment (from 80 to 91%), whereas

M. aff. americana elicited lower numbers (from 100

to 96%) (Figure 3). Aplysina fulva elicited later

successful settlements than the remaining sponges

of the group (except H. melana): between 70 and 80

min during the first experiment (Figure 2), and

from about 5 h (53%) to 8 h (40%) during the

second experiment (Figure 3). Similar results were

obtained in the two experiments with Haliclona

576 C. J. A. de Campos et al.

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melana, which elicited settlements after:3 h and

13% of actinulae settled in the first experiment

(Figure 2) and 7% in the second experiment

(Figure 3). Amphimedon viridis confirmed high

lethal activity (100% of the actinulae died within

10 min of exposure).

Figure 2. Cumulative percentage of actinulae of Zyzzyzus warreni settled on sponges during the first experiment.

Selection of substrata by Zyzzyus warreni 577

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Based on these observations, four settlement re-

sponses were recognized: (a) high and short-time

larval settlement, in actinulae exposed to H.

cebimarensis, M. angulosa, M. aff. americana, M.

laxissima and T. ignis; (b) moderate and delayed larval

settlement, in actinulae exposed to A. caissara,

A. fulva, H. melana and M. microsigmatosa; (c) no

settlement, in actinulae exposed to the algae

H. musciformis and S. cymosum, and to the sponge S.

aurantiacus; and (d) lethal response, in actinulae

exposed to A. rosea, A. viridis and D. reticulatum.

Metamorphosis

During the first minutes of permanent attachment to

preferred substrata, the tentacle tips of the actinulae

adhered temporarily to the surface of the sponges

(Figure 1d). After this adhesion, the aboral protru-

sion emerged from the actinular body, enlarged

proximally presumably via endodermic thickening

and penetrated the sponge tissue. From this enlarged

portion, the hydrocaulus extended and differentiated

upwards, whereas the hydrorhiza extended down-

wards (Figure 1e,f). Three-hour polyps had two

Figure 3. Cumulative percentage of actinulae of Zyzzyzus warreni settled on sponges during the second experiment.

578 C. J. A. de Campos et al.

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classes of stenoteles, two classes of euryteles, and

desmonemes.

During the first 20 h of settlement, the hydro-

caulus increased from 307�356 mm long at 3 h to

1007�1680 mm long at 12 h (Figure 4). An inverse

trend was observed in the hydranth, which de-

creased in length from 479�610 mm at 3 h to 248�337 mm at 12 h. During this period, the aboral

tentacles decreased from 46�89 mm wide at 1 h to

35�55 mm wide at 20 h. Oral tentacles became

externally visible in the hypostome region of the

hydranth:10 h after the beginning of metamor-

phosis (Figure 1f), initially as four small dermic

evaginations that continued to grow out, reaching

98�157 mm long and 29�53 mm wide in 20 h.

Within 20 h, the aboral tentacles reached 246�646

mm long. Most of the differentiation occurred

during this time, the hydranth becoming gradually

vasiform near the base and narrowed in the region

of the hypostome. At:27 h after the beginning of

metamorphosis, the whorl of slightly capitate oral

tentacles and the whorl of filiform aboral tentacles

were observed; meanwhile, endodermal canals were

visible in the hydrocaulus. At:30 h after the

beginning of metamorphosis, the mouth opened

and the polyp was able to search for prey

(Figure 1g). The aboral tentacles were resorbed,

underwent restructuring and lost terminal knobs.

Newly settled polyps occasionally developed new

aboral tentacles.

When sponges were removed from Petri dishes,

primary polyps continued development of a tubular,

thin-walled hydrorhiza covered by filmy perisarc,

often longer than the hydrocaulus itself. Hydrorhi-

zal structures developed until 36 h after metamor-

phosis.

In the laboratory, polyps did not survive until the

development of reproductive structures.

Feeding behaviour

Two-day-old polyps had the ability to search actively

for prey. Fragments of nauplii were caught by aboral

tentacles and taken into the mouth through the oral

tentacles. Foraging for prey consisted of two main

behavioural patterns: (1) a passive pattern, charac-

terized by non-moving, upright posture of the polyp

and slow upward/downward movements of aboral

tentacles, by means of which prey and particles were

trapped, retained or intercepted; and (2) an active,

most predominant pattern, characterized by twisting

and bending of the polyp in different directions, by

means of which prey were transported to the feeding

area, caught by aboral tentacles and delivered to the

mouth. In both patterns, aboral tentacles joined with

oral tentacles to form a ‘bridge’ by which prey was

conducted into the mouth. Actinulae were never

observed to use oral tentacles to capture prey.

Swallowing of an Artemia nauplius by adult polyps

(Figure 1h) took approximately:1 min.

Discussion

Most studies on life history of species of Acharadria

and Tubularia have mentioned the existence of one

whorl of oral tentacles (e.g. Pyefinch & Downing

1949; Hawes 1955; Nagao 1960, 1965; Widersten

1968; Yamashita et al. 1997) and a mouth (Zamponi

& Correa 1988) in newly liberated actinulae. The

actinulae of Zyzzyzus warreni do not contain any of

these structures at this stage. They are developed at a

later stage when successful settlement near conspe-

cifics is achieved. In addition, the actinulae of

Z. warreni are larger than those of other tubulariids

(cf. Zamponi & Correa 1988; Yamashita et al. 2003).

We regard these traits as pre-requisites for a short

planktonic life span and maintenance of larval

competence during metamorphosis. This condition,

Figure 4. Morphometrics of Zyzzyzus warreni settled on the sponge Mycale angulosa during the first 20 h of metamorphosis (n�30).

Selection of substrata by Zyzzyus warreni 579

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named as metamorphic competence by Hadfield

et al. (2001), is characteristic of many marine

invertebrate larvae (e.g. cnidarian planulae, mollus-

can veligers, barnacle cyprids).

The morphological changes that take place during

settlement and early metamorphosis have been

described in some detail for some tubulariid species

(Pyefinch & Downing 1949; Berrill 1952). These are

physiologically mediated by changes in nematocyst

composition and dynamics (Kawaii et al. 1997;

Yamashita et al. 2003). In Z. warreni, these processes

involve the development of juvenile structures (mor-

phological changes in the hydranth, elongation of the

hydrocaulus and, at a later stage, development of

oral tentacles) and increase in size of stenoteles.

These changes represent adaptations of this species

to enhance settlement in the preferred substrata

during the period of larval competence.

The means of attachment to substrata are a

character traditionally used for genera distinction

in the Tubulariidae (see Petersen 1979, 1990;

Marques & Migotto 2001). The morphological

diversity of hydrorhizal processes in polyps of Z.

warreni described by Campos et al. (2007) and the

development of a tubular hydrorhiza observed in this

study demonstrate that this phenotype is determined

by the substratum-derived factors discussed above.

Tubulariids have developed a variety of associa-

tions with animals and plants, namely gorgonians

(Silveira & Migotto 1984; Puce et al. 2008), poly-

chaetes (Tardent 1980), barnacles and mussels

(Migotto & Marques 1999) and seagrasses (Nagao

1960). Acharadria japonica, Hybocodon prolifer and

species of Zyzzyzus have been found almost exclu-

sively on sponges (see Hirohito 1988; Petersen

1990). However, many of these associations are not

obligatory. For instance, A. larynx may settle on

species of Enteromorpha, Ectocarpus and Laminaria,

this preference often dependent on the local density

of these substrata (Pyefinch & Downing 1949).

Under artificial conditions, however, A. crocea and

A. larynx are able to settle on surfaces free from

biogenic substrata after delayed settlement (see

Pyefinch & Downing 1949; Kawaii et al. 1997;

Yamashita et al. 1997). In contrast, Z. warreni occurs

only on demosponges in Brazil and other areas, e.g.

in Bermuda on Tedania ignis (Calder 1988); in the

Santa Marta coast of Colombia, on Tedania sp.

(Bandel & Wedler 1987); in Sagami Bay Japan, on

Ceraochalina differentiata, Callyspongia fibrosa,

Haliclona clathrata, H. permollis and Clathria

fasciculata (Hirohito 1988). In our studies, none of

the actinulae of Z. warreni settled on algae or on

surfaces free of biogenic substrata. Most sponges

elicited either settlement or death of the actinulae. In

experiments with the sponge Suberites aurantiacus

and the algae Sargassum cymosum and Hypnea

musciformis there were neither settlement nor death

of the actinulae. Although S. cymosum is often found

in association with the sponges Mycale angulosa

(Duarte & Nalesso 1996) and M. laxissima (pers.

obs.), Z. warreni is never found on the algae.

There are approximately 120 species of demos-

ponges recorded in the Sao Sebastiao Channel

(Hajdu et al. 2002). The sponges used in this study

belong to the group of the 25 most frequently found

sponges in the channel (see Santos & Hajdu 2001).

Of these, Mycale angulosa, M. laxissima, M.

microsigmatosa and Tedania ignis are preferred sub-

strata used by the hydroid in the natural environ-

ment. These soft sponges constitute a significant

proportion of the biomass available for actinular

settlement and vary considerably with respect to

their encrusting and massive habits (see Lavrado

2006), which facilitates tissue penetration by newly

settled polyps. The lack of association between

Z. warreni and Halichondria cebimarensis, Aplysina

caissara, A. fulva and M. aff. americana could be

explained by the combined effect of abiotic and

biotic factors. For instance, H. cebimarensis is a rare

species in the Sao Sebastiao Channel; the typical

areas of occurrence of A. caissara are highly energetic

rocky coasts (Pinheiro & Hajdu 2001); A. fulva is

occasionally associated with the zoanthid Palythoa

caribaeorum (Pinheiro & Hajdu 2001) and M. aff.

americana is often found underneath rocks. These

constitute unfavourable conditions for settlement of

Z. warreni. In contrast, the diverse and abundant

fauna of crustaceans, polychaetes, molluscs and

hydroids known to live in association with

M. angulosa and M. microsigmatosa (Duarte &

Nalesso 1996; Ribeiro et al. 2003) do not prevent

colonization of Z. warreni on these sponges. In fact,

ophiuroids and polychaetes associated with M. aff.

americana were observed to prey upon actinulae of

Z. warreni during laboratory experiments. The

association of H. cebimarensis with ophiuroids, tube-

worms, ectoprocts, Sargassum and calcareous algae

(Carvalho & Hajdu 2001) could affect the successful

settlement of Z. warreni.

It was outside the scope of this study to evaluate

the role of physical or chemical stimuli during

settlement and metamorphosis of Z. warreni on

biogenic substrata. We hypothesize, however, that

chemical signalling from sponges plays a significant

role in determining the distribution of this hydroid.

From an evolutionary point of view, Puce et al.

(2005, p. 78) noted that associations between

hydroids and sponges ‘evolved as a form of parasit-

ism, in which some hydroid species succeeded in

overcoming the chemical defences that prevented the

settlement of other organisms on most sponges’.

580 C. J. A. de Campos et al.

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The death of actinulae of Z. warreni exposed to

Amphimedon viridis, Aplysilla rosea and Dragmacidon

reticulatum could be associated with allelopathic

defensive mechanisms exhibited by these species

(see Berlinck et al. 1996; Muricy & Silva 1999).

Furthermore, sponges kept in laboratory conditions

often disintegrate rapidly. It is likely that some of the

toxic reactions observed were due to decaying

sponge tissues resulting from the in-vitro conditions.

High cytotoxic, antifungicidal, antiviral and antibac-

terial activities have been identified in the halitoxin

and amphitoxin complex isolated from A. viridis

from Brazil (Berlinck et al. 1996). Cytotoxic activity

has been also identified in extracts of A. rosea

(Berlinck et al. 2004) and A. fulva (Aiub et al.

2006). Similarly, Nunez et al. (2008) found

dibromotyrosine-derived compounds in A. fulva.

Biotoxic activity has not been found in D.

reticulatum, M. aff. americana and Haliclona melana

from Brazil (e.g. Sole-Cava et al. 1981; Muricy et al.

1993; Rangel et al. 2001). Dresch et al. (2005)

tested 20 species of sponges for the presence of

lectinic and haemolytic activity and did not find any

of these activities in Tedania ignis collected from

Santa Catarina State. However, high cytotoxic

activity has been detected in this species from

Florida Keys (Schmitz et al. 1983). It should be

noted that allelopathic defensive mechanisms may

vary significantly with the geographical location of

individual populations. As noted by Nunez et al.

(2008), the composition of secondary metabolites in

sponges can be influenced by multiple factors, such

as habitat, exposure to predation, association with or

infection by microorganisms, individual biosynthesis

or even laboratory techniques used for the purposes

of extraction and purification of compounds.

In summary, our study shows that polyps of Z.

warreni are not able to maintain an independent

existence from sponges. The establishment of the

hydroid�sponge pairing results from considerable

species discrimination by the actinulae during their

competent stage. Further studies are required to

understand the role of chemical signalling on this

selective advantage.

Acknowledgements

We thank Marcio Custodio (IB � USP) for his

constructive comments during this investigation,

and to Stefania Puce and an anonymous referee for

reviewing and improving an early version of the

manuscript. A. E. Migotto and A. C. Marques

received financial support from the Conselho Na-

cional de Desenvolvimento Cientıfico e Tecnologico

(CNPq; 305608/2006-1, 481309/2007-1, 302596/

2003-8, 304720/2009-7, and 562143/2010-6) and

FAPESP (1998/07090-3, 2001/02626-7, 2003/

02432-3, 2004/09961-4, 2006/05821-9, and 2011/

50242-5). U. Pinheiro received financial support

from Fundacao de Amparo a Ciencia e Tecnologia

do Estado de Pernambuco.

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