Research Article |
Corresponding author: Giuseppe Montesanto ( giuseppe.montesanto@unipi.it ) Academic editor: Stefano Taiti
© 2018 Giuseppe Montesanto.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Montesanto G (2018) Presence of a stridulatory apparatus in the manca stages of isopods (Crustacea, Isopoda, Oniscidea). In: Hornung E, Taiti S, Szlavecz K (Eds) Isopods in a Changing World. ZooKeys 801: 501-518. https://doi.org/10.3897/zookeys.801.23018
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Armadillo officinalis Duméril, 1816 (Armadillidae) is a widespread terrestrial isopod species in the Mediterranean basin and on the western coasts of the Black Sea. The species is adapted to live in xeric environments and has mainly nocturnal habits. This species is capable of producing stridulations, which is nowadays recognized as a synapomorphy of the genus. In both sexes, these vibrations are produced by a line of scales on the propodus of pereopod 4 and 5. The main goals of this study are: to describe the manca stages of Armadillo officinalis; to detect the presence of the stridulatory apparatus in the manca stages; to evaluate the differences of such apparatus in the various manca stages. The manca stages (I, II, III) of Armadillo officinalis are described for the first time showing: i, the shortest duration (known in literature) of the manca stage I (approximately 30 minutes); ii, the presence of a rudimental stridulatory organ that may be of great importance in terms of evolutionary aspects and adaptation to terrestrial life. Notes on the reproductive biology are also reported. Furthermore, some considerations on future perspectives for A. officinalis as a model species in biotremology are also discussed.
Armadillo officinalis , biotremology, crustaceans, manca stages, terrestrial isopods
Armadillo officinalis Duméril, 1816 is a species of terrestrial isopod (Crustacea, Isopoda, Oniscidea) belonging to the family Armadillidae. The genus Armadillo Latreille, 1802 is restricted to the Mediterranean basin and western Asia (
In the last years, A. officinalis has been the object of several studies in different fields of ethology, ecology, and reproductive biology (
The feeding preferences and the duration of the stages and substages of the moult cycle of this species was recently studied in detail by
Although many of its biological features are well known, the post marsupial manca stages have never been described. The aims of this study are: to describe the manca stages of Armadillo officinalis; to detect the presence of the stridulatory apparatus in the manca stages and to describe it in these different stages.
With the aid of forceps, numerous specimens of Armadillo officinalis were collected, under the stones of Catania University campus, eastern Sicily (DMS: 37°31'39"N 15°04'20"E); they were then bred in Pisa (western Tuscany), in a climate room at 20°C, with a natural photoperiod. Ovigerous females were separated from the main livestock and bred separately. Each ovigerous female was kept in a periodically moistened Petri dish (Falcon® 351029, 100×15mm, with plaster of Paris substrate), and fed with slices of potatoes and plane-tree leaves (as in
Once the mancas were released from the marsupium they were counted, separated from the female, and raised individually in Petri dishes. The ovigerous females were daily monitored, in order to record the time of manca release. In the same way the postmarsupial mancas were observed every day, so that the moulting process and the time of each manca stage could be monitored and recorded. Twenty-five ovigerous females were dissected in order to study the first manca stage in its intramarsupial development.
Throughout postmarsupial development, twenty individuals, representing each postmarsupial manca stage, were fixed in 70% ethanol for a later analysis. The manca stages were described in accordance with the previously described procedures (
In order to evaluate the number of mancas released from the marsupium, thirty-two ovigerous females of Armadillo officinalis were considered. Fig.
Armadillo officinalis showed three postmarsupial stages (called manca stages M I, M II, and M III), separated by ecdysis and generally characterized by the absence of the first pleopods and non-fuctional seventh pereopods. The following three main characteristics can be used to distinguish each stage: the length of the antennal flagellum articles, the number of ommatidia, the developmental level of the pereopod 7, the presence of the epimera of pereonite 7 (Fig.
Armadillo officinalis Duméril, 1816. A manca stage M I, antennal flagellum B manca stage M II, antennal flagellum C manca stage M II, aesthetascs on the second article of antennal flagellum D manca stage M III, antennal flagellum E manca stage M II, eye F manca stage M III, eye G manca stage M I, p7: pereonite 7 H manca stage M II, p7: pereonite 7 I manca stage M III, p7: pereonite 7, e: epimeron. Scale bars: 10 μm (C, E); 50 μm (A, B, D, F); 100 μm (G, H); 500 μm (I).
Manca stage M I. The duration of this stage varied from a minimum of 21 to a maximum of 43 min, with a mean value of 28 min. The mean body length was 1.76 mm (SD: ± 0.10), with a range from 1.69 to 1.89 mm. The mean cephalothorax width was 0.40 mm (SD: ± 0.07), with a minimum of 0.39 mm and a maximum of 0.46 mm. All the mancas of the first larval stage emerge from the marsupium during the anterior ecdysis process (number of females observed: N = 16) or during the posterior ecdysis (N = 4). No females were observed while giving birth of mancas M I during premoult or intermoult stages. During the ecdysis the mancas remain under their mother’s body and quickly eat both the posterior and anterior exuviae. These mancas had no pigmentation, except for the ommatidia and little brown spots on the pereonite margins; the calcification of the cuticle seems to be incomplete. Because of their body transparency it was possible to observe the exuviae inside the gut. Dorsal surface without scale-setae (see also Fig.
Armadillo officinalis Duméril, 1816. Manca stage M I. A body, lateral view (scale bar 0.5 mm) B cephalothorax, dorsal view C cephalothorax, frontal view D antennula E antenna F left mandible G right mandible H maxillula I maxilla J maxilliped K pereopod 1 L pereopod 2 M pereopod 3 N pereopod 4 O pereopod 5 P pereopod 6 Q pleopod 2 exopod R pleopod 3 exopod S pleopod 4 exopod T pleopod 5 exopod U left uropod V telson.
Stridulatory apparatus of Manca stage M I. Presence of a line of 28–30 scales (plectrum) of approx. 100 μm on sternal margin of pereopod 4 and 5 propodus (Fig.
Armadillo officinalis Duméril, 1816. Manca stage M I. A pereopod 4 (p4) and pereopod 5 (p5) showing the line of scales on the propodus B pereopod 5 propodus, sternal view C the line of scales on the pereopod 4 propodus D scales of the plectrum on the pereopod 5 propodus. Scale bars: 1 μm (D); 10 μm (C); 50 μm (B); 100 μm (A).
Manca stage M II. The duration of this stage varied from a minimum of six to a maximum of seven days, with a mean value of 6.5 days. The mean body length was 2.01 mm (SD: ± 0.04), with a range from 1.92 to 2.11 mm. The mean cephalothorax width was 0.43 mm (SD: ± 0.06), with a minimum of 0.42 mm and a maximum of 0.45 mm. These larvae showed pigmentation on the cephalothorax and the posterior margins of pereonites and pleonites; the calcification of the cuticle seemed to be complete after the previous ecdysis. Even if the body was not completely transparent, a food presence could be observed in the gut as a dark area. At this stage, the mancas left the mother in search of food. Presence of dorsal scale-setae on the body surface (see also Figs
Armadillo officinalis Duméril, 1816. Manca stage M II. A body, lateral view (scale bar 0.5 mm) B dorsal scale-seta C cephalothorax, dorsal view D cephalothorax, frontal view E antennula F antenna G left mandible H right mandible I maxillula J maxilla K maxilliped L pereopod 1 M pereopod 2 N pereopod 3 O pereopod 4 P pereopod 5 Q pereopod 6 R pleopod 2 exopod S pleopod 3 exopod T pleopod 4 exopod U pleopod 5 exopod V left uropod W telson.
Stridulatory apparatus of Manca stage M II. The presence of a line of 38–40 scales (plectrum) of approx. 165 μm on sternal margin of pereopod 4 and 5 propodus could be observed (Fig.
Armadillo officinalis Duméril, 1816. Manca stage M II. A pereopod 4 (p4) and pereopod 5 (p5) showing the line of scales on the propodus B epimera of pereonite 4 (e4), pereonite 5 (e5), and pereonite 6 (e6), ventral view C epimera of pereonite 5, ventral view D epimera of pereonite 6, ventral view. Scale bars: 50 μm (A, C, D); 100 μm (B).
Manca stage M III. The duration of this stage varied from a minimum of 20 to a maximum of 24 days, with a mean value of 22 days. The mean body length was 2.35 mm (SD: ± 0.09), with a range from 2.16 to 2.60 mm. The mean cephalothorax width was 0.56 mm (SD: ± 0.08), with a minimum of 0.51 mm and a maximum of 0.59 mm. These larval stage showed more pigmentation than in the previous stage, on the cephalothorax, pereonites, and pleonites; the calcification of the cuticle is complete after the previous ecdysis. Higher presence of dorsal scale-setae on the body surface (see also Fig.
Armadillo officinalis Duméril, 1816. Manca stage M III. A body, lateral view (scale bar 0.5 mm) B cephalothorax, dorsal view C cephalothorax, frontal view D antennula E antenna F left mandible G right mandible H maxillula I maxilla J maxilliped K pereopod 1 L pereopod 2 M pereopod 3 N pereopod 4 O pereopod 5 P pereopod 6 Q pleopod 2 exopod R pleopod 3 exopod S pleopod 4 exopod T pleopod 5 exopod U left uropod V telson.
Stridulatory apparatus of Manca III stage. Also at this stage it was possible to observe the presence of a line of approx. 40 scales (plectrum) of 160 μm on sternal margin of pereopod 4 and 5 propodus (Fig.
Armadillo officinalis Duméril, 1816. Manca stage M II. A pereopod 4 (p4) and pereopod 5 (p5) showing the line of scales on the propodus B scales of plectrum of pereopod 4 propodus C epimera of pereonite 5 (e5), pereonite 6 (e6), and pereonite 7 (e7), ventral view. Scale bars: 5 μm (B); 50 μm (A); 100 μm (C).
This detailed study of the morphology of the manca stages of Armadillo officinalis has highlighted their differences. The cephalothorax develops a complete frontal line, passing from M I to manca stage M II. In the antennula the number of apical aesthetascs varies from five on M I to seven on M III. Antennal flagellum changes the proportion of the two articles from M I to M II; the aesthetascs appear on the flagellum in M II and remain the same in M III. Buccal appendages do not modify their main structure in the three manca stages. The number of setae in pereopods increased from M I to M III; pereopod 1 shows a well-defined hairy area on carpus from manca stage M II, pereopods 4 and 5 propodus show a line of scales in the three manca stages, but the number of the scales and their distance on the sternal margin of propodus varies distinctly; pereopods 7 are absent in the first manca stage, there are hints in the second stage, and they are fully developed but ventrally folded in the third manca stage. Pleopods 1 are absent in manca stages M I and M II, but appear in the third manca stage as a hint. Setae on pleopod 2–5 exopods appear in the manca stages M II and they do not considerably increase their number in the next stage. In the three manca stages, the uropod and telson change substantially in shape and proportion.
As for the comparison with other larval stages of terrestrial isopods, the mancas of A. officinalis show some significant differences. First of all, as reported for Atlantoscia floridana (Van Name, 1940) (
The main body development follows the same rules of the already known oniscidean manca stages: the proportion inversion of the antennal flagellum articles, the development of the seventh pereonite, the appearance of pereopod 7 and pleopod 1. Other minor differences are in the number of ommatidia, and in the number of setae on the margins of pereopods and pleopods (
The present study has also shown the presence of the stridulatory apparatus, even in A. officinalis early stages of development, which is an absolutely new issue for the taxon Oniscidea. The presence of similar apparatus in different taxa of Arthropoda (especially in Insecta) was widely known, even in larval or juvenile stages (
The presence of a such stridulatory apparatus definitely is a synapomorphic character of the genus Armadillo. The identification of the typical stridulatory structure of adults in the mancas, even in an early stage development, represents a relevant discovery. Its presence must be, without doubt, the result of a long evolutionary process. It leads to believe that this character, defining the genus Armadillo (sensu
It still remains an open question the purpose of a stridulatory apparatus in larval stages, even in biological circumstances in which it could not be used (e.g. inside the maternal brood pouch). I have observed that adults of A. officinalis produce stridulation only when their body is rolled up in a ball: this should be a further step in an hypothetic line of a defense strategy or aggregation phenomena. Further studies on these aspects are currently underway. The presence of stridulatory apparatus in larvae of other group of arthropods is well known.
Moreover, other fields of research investigate the possibility of A. officinalis to produce substrate-borne vibrations instead of air-borne sounds, as defined in
Science already knows approximately 3,800 species of terrestrial isopod (
With regard to the findings in the present study, the presence of a stridulatory apparatus in a terrestrial crustacean surely constitute an important issue on the terrestrialisation of this taxon (see also
I would like to acknowledge Prof. Franco Verni for his great willingness and Dr. Simone Gabrielli for the assistance in the realization of the SEM photographs (Centro Interdipartimentale di Microscopia Elettronica, Università di Pisa). I am also grateful to Prof. Paula B. Araujo (Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil), Prof. Spyros Sfenthourakis (University of Cyprus, Nicosia, Cyprus), Prof. Jasna Štrus (University of Ljubljana, Ljubljana, Slovenia), and Prof. Ivan H. Tuf (Palacký University, Olomouc, Czech Republic) for expressing their interest in this research project during the 10th International Symposium on the Biology of Terrestrial Isopods (ISTIB), Budapest, Hungary (August, 2017), and for their inspiring words that suggested me many improvements of this research. I am very much thankful to Dr. Stefano Taiti (ISE-CNR, Florence, Italy) for his constant useful advice and suggestions. Finally, I would like to acknowledge the anonymous reviewers for their precious comments and Dr. Annamaria Pulina (University of Pisa) for English language revision.