Bull. Eur. Ass. Fish Pathol., 30(6) 2010, 220

Larval anisakid nematodes of Histioteuthis reversa (Verril, 1880) and H. bonnellii (Férussac, 1835) (Cephalopoda: Teuthoidea) from Sardinian Channel (western Mediterranean)

J. Culurgioni, D. Cuccu, M. Mereu and V. Figus*

Dipartimento di Biologia animale ed Ecologia, Viale Poeo 1, 09126 Cagliari, Italy

Abstract Larvae of two Anisakidae nematodes from the Histioteuthidae caught in Sardinian Channel (western Mediterranean) are described. Third-stage larvae of Lappetascaris sp. Type A (sensu Nagasawa and Moravec, 2002) measured about 16-28 mm long and occurred mainly encysted, not encapsuled, in the mantle musculature or free in the mantle cavity of the 4.26% of Histioteuthis bonnellii (Férussac, 1835) and of the 3.05% of H. reversa (Verril, 1880) examined. Larvae of Anisakis Type II (sensu Berland, 1961) were about 28-29 mm long and were found only in H. bonnellii (P%=1.83), free in the mantle cavity or adhered to the stomach wall. These findings represent new host reports for both Lappetascaris sp. Type A and Anisakis Type II larvae in Histioteuthidae, and new geographical record for the genus Lappetascaris Rasheed, 1965 in Mediterranean waters.

Introduction The Histioteuthidae (Cephalopoda: allowing the transmission of parasites in the Teuthoidea) is a mesopelagic family inhabiting marine ecosystem. In fact, as cephalopods, all the world oceans composed of 1 genus with they work as intermediate, paratenic or 13 species. (Voss et al., 1998). reservoir hosts in the nematode life cycle (Oshima, 1972; Hochberg, 1983, 1990; Pascual In the Mediterranean Sea, as reported by Voss et al., 1995; Pascual et al., 1996). et al. (1998) this family is represented by 2 species, the reverse jewel squid Histioteuthis To date, reports on nematode parasites of reversa (Verril, 1880) and the umbrella squid Mediterranean Histioteuthidae are restricted H. bonnellii (Férussac, 1835) inedible for to an unidentified larva found in H. bonnellii humans. Although this, their preference from France by Dollfus (1958). for high productive waters ascribes them a key role in the marine food chain both as During a study concerning the biology and predators (Oshima et al., 1969; Hochberg, distribution of cephalopod stocks in Sardinian 1983) and preys (Bello, 1991, 1996; Voss et al., waters (Cuccu et al., 2007), parasitological 1998; Santos et al., 1999; Xavier et al., 2007), examinations were carried out.

* Corresponding author’s e-mail: vfi[email protected] Bull. Eur. Ass. Fish Pathol., 30(6) 2010, 221

This paper provides an important integration and sex and gonad maturity stage were in the knowledge of parasites of Mediterranean assessed (Voss et al., 1998). Then, the squids cephalopods indicating a new host for the were examined for parasites using standard nematode Anisakis sp. Type II (sensu Berland, diagnostic techniques. 1961) and new hosts and a new geographical record for the genus Lappetascaris Rasheed, The larval nematodes detected were collected 1965. and counted, then washed in physiological saline and observed as fresh mounts, fixed Materials and methods in 70° ethanol and cleared in lactophenol. Samples of 141 H. reversa and of 164 H. bonnelli Figures were made with a drawing tube were collected between April 2005 and April and a Nikon digital camera connected to an 2009 in the Sardinian Channel (western Olympus microscope. All measurements are Mediterranean). Trawlings were conducted presented in micrometers (μm). during commercial bottom fishing, in 7 different hauls comprised between 38°36’ and The parasitological indices as Prevalence (P%), 39°07’ N and 8°13’ and 9°31’ E (Figure 1), at a Mean Intensity (MI), and Intensity range were depth range of 550-700 m. calculated according to Bush et al. (1997).

Immediately aVer landing, in the laboratory, Results dorsal mantle length (DML) and total body Six out 141 H. reversa examined (P%=4.26) weight (TBW) for each fresh specimen were were found infected by third-stage anisakid taken according to Roper and Voss (1983), larvae of the genus Lappetascaris Rasheed,

Figure 1. Map of the Sardinian Channel (western Mediterranean). a-f, hauls in the fishing area for H. reversa and H. bonnellii. Bull. Eur. Ass. Fish Pathol., 30(6) 2010, 222

1965 (Raphidascardiinae). The infected squids reversa and in H. bonnellii, and by Anisakis sp. harboured 1 to 9 worms with a mean intensity Type II in H. bonnellii, occurred only in sex- of 3.8. The larvae were detected free in the determined specimens, without sex specificity, mantle cavity or encysted (not encapsuled) in while no infection were observed in young, the mantle musculature (Figure 2). sex-undetermined squids.

Regarding H. bonnellii, 5 out 164 examined Description specimens (P%=3.05) harboured third-stage Family: Anisakidae Railliet and Henry, 1912 larval nematodes. In detail, Lappetascaris Subfamily: Raphidascaridinae Hartwich, 1954 sp. larvae were observed in 2 H. bonnellii Genus: Lappetascaris Rasheed, 1965 (P%=1.22), respectively infected by 1 and 13 Lappetascaris sp. Type A (sensu Nagasawa and worms, free or encysted within the mantle Moravec, 2002) (Figure 4) (MI =7). Third-stage larvae (based on 11 specimens): In 3 umbrella squids, larvae morphologically body elongated and whitish, 20183 (15688- identified as Anisakis sp. Type II (sensu Berland, 27707) μm long; maximum body width 364 1961) were detected (P%=1.83). These larvae (258-481) μm; cephalic end narrowed and were free in the mantle or strongly adhered to obliquely truncated, so that the ventral side the external viscera wall (Figure 3) with mean protrudes more than dorsal one. Primordia intensity of 1.67 (range 1-3). of lips anteriorly covered by cuticolar coating and laterally supported by a sclerotized Both the infections by Lappetascaris sp. in H. structure supplied with posterior processes,

Figure 2. Lappetascaris sp. larva in the mantle of H. reversa from Sardinian Channel (western Mediterranean). Bull. Eur. Ass. Fish Pathol., 30(6) 2010, 223

Figure 3. Anisakis sp. Type II larvae encysted in the visceral wall of H. bonnellii. ventral side of the whole structure 73 (65-88) reversa, reverse jewel squid (Cephalopoda: μm long. Oesophagus cylindrical, 1883 (1427- Teuthoidea: Histioteuthidae). 2920) μm long; oesophageal gland distinct; Site of infection: mantle musculature, mantle ventriculus oval, 243 (172-373) μm long and cavity. 120 (74-156) μm wide. Intestinal caecum Locality: Mediterranean Sea: Sardinian extending always behind the level of excretory Channel (haul d: 38°55’N and 9°21’E / 39°02’N pore, 1633 (1060-2533) μm long; ventricular and 9°16’E, 9 Dec 2005, 22 Jun and 12 Oct appendix very long, measuring 13129 (8970- 2007, 11 Jan 2008, 4 Feb 2008 ― H. reversa, 18 19951) μm; intestine straight, rectum hyaline Jan 2008 ― H. bonnellii; haul b: 38°36’N and with three drop-shaped rectal glands. Nerve 8°35’E / 38°42’N and 8°51’E, 7 Apr 2009 ― H. ring situated 369 (278-421) μm from anterior bonnellii). extremity; distance between excretory pore and anterior extremity 461 (411-495) μm; tail Comments conical with slightly inflated extremity, 255 The nematodes collected correspond in their (179-377) μm long, bearing a caudal spike morphology to the third-stage larvae of apparently formed by two segments, 7.1 (6.5- Lappetascaris sp. described first by Nagasawa 8.1) μm long. and Moravec (1995) in the common squid Todarodes pacificus (Steenstrup, 1880) from Representative specimens are deposited in the the Sea of Japan, and to Lappetascaris sp. Museo di Zoologia del Dipartimento di Biologia Type A found by the same authors (2002) Animale ed Ecologia (University of Cagliari). in Thysanoteuthis rhombus Troschel, 1857, Ommastrephes bartramii (Lesueur, 1821), Report details Onychoteuthis borealijaponica Okada, 1927, and Host: H. bonnellii, umbrella squid and H. Gonatopus borealis Sasaki, 1923 from the central Bull. Eur. Ass. Fish Pathol., 30(6) 2010, 224

Figure 4. Third-stage larvae of Lappetascaris sp. from Histioteuthis reversa from Sardinian Channel (western Mediterranean). A, anterior part of body; B, general view; C, cephalic end; D, caudal end. cc, cuticolar coating of the lips primordia; cs, caudal spike; ep, excretory pore; i, intestine; ic, intestinal caecum; nr, nerve ring; oe, oesophagus; pl, primordium of lips; ps, process of sclerotized support of lips primordia; r, rectum; rg, rectal glands; ss, sclerotized support of lips primordia; v, ventriculus; va, ventricular appendix. Bull. Eur. Ass. Fish Pathol., 30(6) 2010, 225 and northern Pacific Ocean. rectum short with three rounded rectal glands. Distance of nerve ring from anterior As reported by Nagasawa and Moravec (1995), extremity 323 (322-324) μm; excretory pore another description of third-stage larvae of the ventral, situated at 48 (48-48) μm from anterior same genus is given by Moravec and Scholz extremity; tail conical, without mucron, 270 in 1991 from the cyprinid fish Paralaubuca cf. (225-314) μm long. typus, but the same authors assert that the two forms belong to different congeneric species. Representative specimens are deposited in the Museo di Zoologia del Dipartimento di Biologia To date, the genus Lappetascaris comprises Animale ed Ecologia (University of Cagliari). only one species, L. lutjanii, first described by Rasheed (1965) from Lutjanus sp. (Bloch, Report details 1790) (Lutjanidae) and Hilsa ilisha (Hamilton, Host: H. bonnellii 1822) (Clupeidae) from Karachi waters Site of infection: stomach wall; mantle cavity. (Pakistan), then detected by other authors in Locality: western Mediterranean: Sardinian marine, brackish and fresh water teleosts from Channel (haul d: 38°55’N and 9°21’E / 39°02’N India, Pakistan (Soota and Dey-Sarkar, 1980; and 9°16’E, 15 Apr 2005, 5 May 2006; haul b: De, 1990) and from Atlantic coasts of Brazil 38°36’N and 8°35’E / 38°42’N and 8°51’E, 3 (Vicente et al., 2002). Apr 2009).

This is the first report of the presence of the Comments genus Lappetascaris both in Mediterranean Sea These larvae differ from those of Anisakis and in the Histioteuthidae. sp. Type I (sensu Berland, 1961) and are characterized by a short ventriculus with Description a horizontal ventriculo-intestinal junction Family: Anisakidae Railliet & Henry, 1912 and a long conical tail without a mucron, Subfamily: Anisakinae Chabaud, 1965 corresponding to Anisakis sp. Type II (sensu Genus Anisakis Dujardin, 1845 Berland, 1961). This larval morphotype was Anisakis sp. Type II (sensu Berland, 1961) reported in Mediterranean Sea from several (Figure 5) species of fish and cephalopods (Maiucci et al., 1986, 2001; Orecchia et al., 1989; Culurgioni Third-stage larvae (based on 2 specimens): et al., 2006). Nowadays, Anisakis sp. Type body length 28552 (28311-28792) μm; II morphotype comprises three genetically maximum body width 831 (828-833) μm; distinct species: A. physeteris Baylis, 1923, A. cuticle transversely finely striated; anterior brevispiculata Dollfus, 1966 (Maiucci et al., extremity rounded, ventral lip with one larval 2001), and A. paggiae (Maiucci et al., 2005). tooth 21.6 (21-22.2) μm long. Oesophagus straight, 1904 (1875-1932) μm long; ventriculus This is the first report of the presence of white, relatively short, 592 (516-667) μm long third-stage larvae of Anisakis sp. Type II in H. and 335 (291-379) μm wide; intestine straight, bonnelli from Mediterranean Sea. Bull. Eur. Ass. Fish Pathol., 30(6) 2010, 226

Discussion Concerning Lappetascaris sp., it is noticeable the first observation of this genus in Mediterranean and, considering the role of cephalopods as intermediate or paratenic hosts in the transmission of Anisakidae, it would be important to deepen the study on the life-cycle of this parasite, identifying its first hosts, probably euphasiids and other shrimps and its definitive hosts. Regarding the laer, since adults of the single species of this genus, L. lutianji, as reported above, has been detected in teleosts both from Indian Ocean and from Atlantic coasts of Brasil (Rasheed, 1965; Soota and Dey-Sarkar 1980; De, 1990; Vicente et al., 2002), it would be interesting to pursue further investigations on histioteuthids-feeding Mediterranean teleosts as Xiphias gladius L. (Bello, 1991, 1996).

It’s known that Anisakis sp. Type II larvae lack morphologically useful characters for an identification to the specific level by light microscopy. On the other hand, geographical data concerning the distribution of A. physeteris, A. brevispiculata and A. paggiae, ascribed to the Anisakis sp. Type II morphotype (Maiucci et al., 2005) suggest that the specimens described in the present work may be A. physeteris, also reported recently by Farjallah et al. (2008) from the Northern Tunisian coasts, i.e. Sardinian Channel. This hypothesis is also supported by the fact that H. bonnellii is one of the most important preys of the sperm whale Physeter Figure 5. Third-stage larvae of Anisakis sp. Type II catodon L. in Atlantic and in Mediterranean from Histioteuthis bonnellii from Sardinian Channel (western Mediterranean). A, anterior part of body; waters (Santos et al., 1999; Roberts, 2003) B, caudal end. i, intestine; nr, nerve ring; oe, well known as the main definitive host of A. oesophagus; r, rectum; rg, rectal glands; vt, ventral tooth; v, ventriculus. physeteris (Maiucci et al., 2001). Bull. Eur. Ass. Fish Pathol., 30(6) 2010, 227

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