Issues of Decapod Crustacean Biology Developments in Hydrobiology 184 Series editor K. Martens Issues of Decapod Crustacean Biology Edited by M. Thessalou-Legaki University of Athens, Greece Reprinted from Hydrobiologia, Volume 557 (2006) 123 Library of Congress Cataloging-in-Publication Data A C.I.P. Catalogue record for this book is available from the Library of Congress. ISBN 1-4020-4599-9 Published by Springer, P.O. Box 17, 3300 AA Dordrecht, The Netherlands Cover illustration: Logo of the Conference Printed on acid-free paper All Rights reserved Ó 2006 Springer No part of this material protected by this copyright notice may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording or by any information storage and retrieval system, without written permission from the copyright owner. Printed in the Netherlands TABLE OF CONTENTS Preface M. Thessalou-Legaki vii Discrimination between the Mediterranean spiny lobsters Palinurus elephas and P. mauritanicus (Crustacea: Decapoda) by mitochondrial sequence analysis R. Cannas, A. Cau, A.M. Deiana, S. Salvadori, J. Tagliavini 1–4 Short Research Note Study of the nucleolar organizer regions in Palinurus elephas (Crustacea: Decapoda) E. Coluccia, A.M. Deiana, R. Cannas, S. Salvadori 5–8 The constructional properties of the exoskeleton of homarid, palinurid, and scyllarid lobsters S.F. Tarsitano, K.L. Lavalli, F. Horne, E. Spanier 9–20 Biological aspects of Medorippe lanata (Linnaeus, 1767) (Brachyura: Dorippidae) from the eastern Ligurian Sea (western Mediterranean) I. Rossetti, P. Sartor, B. Francesconi, M. Mori, P. Belcari 21–29 Review Paper Reproductive investment in Brachyura R.G. Hartnoll 31–40 An ultrastructural and histochemical study of the germinal cells contained in hemispermatophores of males of the Aristaeomorpha foliacea (Risso, 1827) S. Desantis, M. Labate, G.M. Labate, P. Maiorano, A. Tursi 41–49 Reproduction strategy of the deep-sea hermit crabs Pagurus alatus and Pagurus excavatus of the Central-Western Mediterranean Sea M. Mura, F. Orru` , A. Cau 51–57 Diel and seasonal changes in the structure of a Decapod (Crustacea: Decapoda) community of Cymodocea nodosa from Southeastern Spain (West Mediterranean Sea) J.E.G. Raso, M.J. Mart´ın, V. D´ıaz, V. Cobos, M.E. Manjo´n-Cabeza 59–68 The use of artificial benthic collectors for assessment of spatial patterns of settlement of megalopae of Carcinus maenas (L.) (Brachyura: Portunidae) in the lower Mira Estuary, Portugal J. Paula, I.C. Silva, S.M. Francisco, A.A.V. Flores 69–77 Feeding habits of Percnon gibbesi (H. Milne Edwards, 1853) in the Sicily Strait V. Puccio, M. Relini, E. Azzurro, L.O. Relini 79–84 The decapod crustacean community of the Guadalquivir Estuary (SW Spain): seasonal and inter-year changes in community structure J.A. Cuesta, E. Gonza´ lez-Ortego´n, A. Rodr´ıguez, F. Baldo´, C. Vilas, P. Drake 85–95 vi Preliminary estimation of the factors influencing the distribution of the midlittoral crab Portumnus lysianassa (Herbst, 1796) N. Chartosia, A. Koukouras, M. Mavidis, M.-S. Kitsos 97–106 Chromatic patterns of the hermit crab Calcinus tubularis related to the occupied shell D. Pessani, T. Tirelli 107–112 Trawl selectivity trials on the deep-water rose shrimp (Parapenaeus longirostris) in Sicilian waters S. Ragonese, M. L. Bianchini 113–119 Catch composition and damage incurred to crabs discarded from the eastern Ligurian Sea ‘‘rapido’’ trawl fishery P. Sartor, B. Francesconi, I. Rossetti, S. De Ranieri 121–133 Fishery of the deep-water rose shrimp Parapenaeus longirostris (Lucas, 1846) (Crustacea: Decapoda) in the northern Tyrrhenian Sea (western Mediterranean) M. Sbrana, C. Viva, P. Belcari 135–144 Selectivity parameters and size at first maturity in deep-water shrimps, Aristaeomorpha foliacea (Risso, 1827) and Aristeus antennatus (Risso, 1816), from the North-Western Ionian Sea (Mediterranean Sea) R. Carlucci, G. DÕOnghia, L. Sion, P. Maiorano, A. Tursi 145–154 Catch composition on red shrimps’ (Aristaeomorpha foliacea and Aristeus antennatus) grounds in the Eastern Ionian Sea Ch. Mytilineou, S. Kavadas, C.-Y. Politou, K. Kapiris, A. Tursi, P. Maiorano 155–160 Hydrobiologia (2006) 557:vii Ó Springer 2006 M. Thessalou-Legaki (ed.), Issues of Decapod Crustacean Biology DOI 10.1007/s10750-005-1300-4 Preface Maria Thessalou-Legaki University of Athens, Greece This volume includes the proceedings of the 8th P. Y. Noe¨ l (France), J. Paula (Portugal), K. Sakai Colloquium Crustacea Decapoda Mediterranea, (Japan), F. Sarda` (Spain), R. Serjidi (Algeria), E. which was held at the Ionian University on Corfu Spanier (Israel), Zd. Sˇ tevcˇ ic´ (Croatia), M. Thes- Isl., Greece, from 2 to 6 September 2002 and was salou-Legaki (Greece), M. Tu¨ rkay (Germany) and organized by the National and Kapodistrian M. Vannini (Italy). University of Athens. The meeting is the oldest The Organizing Committee, based at the European carcinological event held regularly at a Department of Zoology–Marine Biology of the 3-year interval. Since 1972, institutions from University of Athens, was composed of M. Apos- Mediterranean countries organized Colloquia in tolopoulou, A. Chilari, K. Kapiris, K. Kevrekidis, Rovinj, Ancona, Barcelona, Thessaloniki, Paris, A. Merakou, S. Papaspyrou, J. Paula and Zd. Florence and Lisbon. Although the meeting is Sˇ tevcˇ ic´ and M. Thessalou-Legaki as coordinator. devoted to the Mediterranean area, contributions Many thanks are due to all the institutions and worldwide are mostly welcome, providing, thus, a persons who helped in the organization of the forum for fruitful contact and scientific exchange. meeting and the publication of the proceedings: In Corfu, 120 delegates from 21 countries rep- the Ionian University, especially Prof. A. Verga- resenting all continents met and presented 8 ple- nelakis and Mrs. E. Kourkoulou, for providing the nary talks, 46 oral and 74 poster presentations. facilities in Corfu; the University of Athens, the The themes included covered all issues of decapod Greek Ministry of Education, the Region of biology: systematics, phylogeny, genetics, ethol- the Ionian Islands and the Municipality of Corfu ogy, morphology, biogeography, reproduction, for supporting the meeting financially or providing ecology, physiology and fisheries. A special session us with facilities and services. I would like to ex- was devoted to deep-water shrimps and related press my gratitude to Hydrobiologia as well as its research projects. previous and present Editors-in-Chief, Profs. H. The Scientific Committee of the 8th Collo- Dumont and K. Martens, for the collaboration, to quium Crustacea Decapoda Mediterranea was the referees for their thorough reviews and to Mrs. composed of K. Anger (Germany), G. Charman- A. Chilari for the support in the preparation of the tier (France), P. Dworschak (Austria), A. Elef- proceedings. Thanks are also due to the members theriou (Greece), W. Emmerson (South Africa), D. of the Organizing Committee, students, family and L. Felder (USA), C. H. J. M. Fransen (The friends for their effective efforts, to the members of Netherlands), C. Froglia (Italy), B. Galil (Israel), the Scientific Committee for their valuable sug- J. E. Garcı´ a Raso (Spain), R. G. Hartnoll (UK), gestions and to all participants and authors of the A. Koukouras (Greece), P. K. L. Ng (Singapore), present volume. Hydrobiologia (2006) 557:1–4 Ó Springer 2006 M. Thessalou-Legaki (ed.), Issues of Decapod Crustacean Biology DOI 10.1007/s10750-005-9301-x Discrimination between the Mediterranean spiny lobsters Palinurus elephas and P. mauritanicus (Crustacea: Decapoda) by mitochondrial sequence analysis 1, 1 1 1 2 R. Cannas *, A. Cau , A.M. Deiana , S. Salvadori & J. Tagliavini 1 Dipartimento di Biologia Animale ed Ecologia, Universita` di Cagliari, Viale Poetto 1, 09126 , Cagliari, Italia 2 Dipartimento di Biologia Evolutiva e Funzionale, Universita` di Parma, Italia (*Author for correspondence: Tel.: +390706758004; Fax: +390706758022; E-mail: 2 species. In order to examine the genetic variability obtained by the CEQ Dye Terminator Cycle at both the intra- and interspecific level, spiny Sequencing Kit (Beckman Instruments Inc., Ful- lobster DNA samples from different Mediterra- lerton, CA, USA) and LCO and HCOI primers, nean localities were analysed. both directions for all sequences. The sequences, In addition, the same mitochondrial region was typically one almost complete strand for each used to examine the phylogenetic relationships reaction, were obtained by an automatic between the two species of Palinurus and other sequencer, CEQ 2000 sequencer (Beckman species of two genera belonging to the family Instrument Inc.). The sequences were aligned with Palinuridae, Panulirus and Jasus. ClustalX (Thompson et al., 1997). Homologous mtCOI sequences from Panulirus and Jasus spe- cies, obtained by Ptacek et al. (2001) and Ovenden Materials and methods et al. (1997) respectively (see GenBank accession numbers therein), were also included in the phy- Adult specimens of P. elephas and P. mauritanicus logenetic analysis. Pagurus longicarpus Say, 1817 were collected by trammel nets and trawls from (Crustacea, Decapoda, Brachyura) sequence was diverse locations of their distribution area during used as outgroup (Genbank accession number NC 1997–2002. In particular, for the common spiny 003058). A homologous mtCOI region (469 bp) of lobster we analysed 10 individuals from the Sar- 25 taxa, including 217 variable sites and 187 par- dinian Sea (both from the western and the south- simony-informative sites, was analysed by neigh- ern coasts), five individuals from the Ligurian Sea bour joining, maximum parsimony and maximum and two from the Ionian Sea. In the case of the likelihood. Neighbour joining trees derived from pink spiny lobster, we examined five specimens the Kimura two-parameter distances were calcu- that were collected off the Sardinian coasts and lated with the Molecular Evolutionary Genetics two specimens from the Atlantic (off the Portu- Analysis (MEGA2) software package (Kumar guese coasts). et al., 2001). Support for nodes was assessed by For each sample, the leg muscle was dissected bootstrap values (1000 replicates). and stored in an excess of 90% ethanol. Total For parsimony analysis the min-mini heuristic DNA was isolated using the standard phenol/ search algorithm in MEGA2 was employed, with chloroform extraction techniques (Sambrook confidence levels evaluated using 100 bootstrap et al., 1989). replicates. All parsimony-uninformative sites were A region of the COI gene (750 bp, size esti- ignored. Maximum likelihood analyses were per- mated by agarose gel electrophoresis) was ampli- formed using Tree-Puzzle 5.0 software package fied by the polymerase chain reaction (PCR) using (Schmidt et al., 2002) using the fast tree search a pair of primers (LCO 5¢-GGTCAACRAAT- algorithm quartet puzzling (QP) that allows anal- CAYAARGATATTGG-3¢, HCOI 5¢-TAAAC- ysis of large data sets (more than 10–15 sequences) TTCAGGGTGRCCAAAAAATCA-3¢) modified and automatically assigns estimations of support from those given in Folmer et al., (1994). A MJ to each internal branch. We tested all the three Research PTC-100 thermal cycler was used with substitution models implemented in Tree-Puzzle: the following cycle profiles according to the the Tamura-Nei model (Tamura & Nei, 1993), the TOUCHDOWN-PCR approach (Don et al., F84 model (Felsenstein, 1984) and the HKY85 1991): 5 min at 95 °C; 3 cycles of 30 s at 95 °C, model (Hasegawa et al., 1985), obtaining concor- 30 s at 52 °C, 1 min at 72 °C; 3 cycles of 30 s at dant results. In this paper only the tree obtained 95 °C, 30 s at 50 °C, 1 min at 72 °C; 3 cycles of with the HKY85 model is shown. 30 s at 95 °C, 30 s at 48 °C, 1 min at 72 °C; 30 cycles of 30 s at 95 °C, 30 s at 46 °C, 1 min at 72 °C; 1 min at 72 °C. Results COI PCR-amplified fragments were purified from TAE agarose gel electrophoresis by the Nucleotide sequences for 659 bp of the mt-COI QIAquick gel extraction kit (Promega, Wisconsin, gene were determined through direct sequencing of USA). Sequences of the Palinurus specimens were amplified DNA for both species (EMBL 3 Nucleotide Sequence Database Accession Num- the NJ and ML trees, but with lower values of bers AJ889577, AJ889578). bootstrap for the clade of Jasus and Panulirus No intraspecific variation was found even when groups III–IV (Fig. 1). comparing sequences obtained from samples from different locations. However, the alignment of the COI sequence Discussion and conclusions permitted to determine several differences between the two species. The P. elephas and P. mauritanicus The taxonomic status within the family Palinuri- sequences present 67 single-base-pair substitutions dae has been recently questioned (Baisre, 1994) (10% of the nucleotide positions); there is no evi- stressing the importance of acquiring genetic dence of insertions or deletions. Nucleotide tran- information on these species in order to address sitions (A to G and C to T=56) greatly outnumber such issues. transvertions (A to T and A to C=11). Third In this paper, DNA nucleotide sequence anal- positions in codons were the most variable (83%) ysis from a region of the mitochondrial genome followed by first positions (13.8%) and only two (the cytochrome oxidase subunit I) was used to changes were observed in second positions. The determine the extent of genetic differentiation COI region was 59% A+T rich, which is similar within and between the two species P. elephas and to the 59.5% and the 57% reported among Jasus P. mauritanicus. and Panulirus species respectively (Ovenden et al., Comparing nucleotide sequences, no intraspe- 1997; Ptacek et al., 2001). Nucleotide content cific differences were observed within our sample averaged 27.1% A, 31.9% T, 18.2% G, 22.8% C. size, not even among individuals collected from The sequence divergence between the two Medi- separate geographic locations. Absence of within terranean species was comparatively low (10%), species variation of the mt-COI gene in Palinurus lower than that reported for Panulirus species species would not exclude the presence of partially (12.4–31.8%) by Ptacek et al. (2001), similar to that calculated between J. caveorum and J. fron- talis, but higher than that calculated for other recognised Jasus species by Ovenden et al. (1997). To investigate the relationships of the Medi- terranean species with other spiny lobster species, phylogenies were produced from mitochondrial sequence variations using our original data and published sequence data (Ovenden et al., 1997; Ptacek et al., 2001). Maximum likelihood (QP) and neighbour-joining (NJ) analyses produced trees with the same overall topology: four major lineages are clearly found; they correspond to the three genera of Jasus, Palinurus and Panulirus (this genus with two sub-lineages corresponding to morphological groups I–II and III–IV of George & Main, 1967). All these four internal branches were supported by moderate to high bootstrap values (73–100%) in NJ analysis and well sup- ported in ML quartet puzzling analysis. Un- Figure 1. Neighbour joining consensus tree based on COI weighted maximum parsimony analysis (MP), partial sequences for the three genera of spiny lobsters: Pal- inurus, Panulirus and Jasus. NJ bootstrap values, in bold, are after 100 bootstrap replicates, yielded two most indicated only for nodes >70%. The same branching pattern parsimonious trees with a length of 1070 steps was obtained with MP and ML quartet puzzling (QP). In circles (consistency index =0.347664, retention index at the principal nodes, the bootstrap support for NJ (bold, 1000 =0.488645, rescaled consistency index=0.16 replicates) and MP (underlined, 100 replicates), as well as the 0689). The consensus parsimony tree is similar to ML QP support values (italic) are given.