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Anisakid Nematodes (Nematoda: Anisakidae) Syst Parasitol (2015) 92:181–195 DOI 10.1007/s11230-015-9597-5 Anisakid nematodes (Nematoda: Anisakidae) from the marine fishes Plectropomus laevis Lace´pe`de (Serranidae) and Sphyraena qenie Klunzinger (Sphyraenidae) off New Caledonia, including two new species of Hysterothylacium Ward & Magath, 1917 Frantisˇek Moravec . Jean-Lou Justine Received: 26 June 2015 / Accepted: 2 August 2015 Ó Springer Science+Business Media Dordrecht 2015 Abstract Based on light and scanning electron presence of narrow lateral alae, a short caecum and a microscopical studies, two new species of Hysterothy- long ventricular appendix, the length (762–830 lm) lacium Ward & Magath, 1917 (Nematoda: Anisaki- and shape of the spicules, the number (37–38 pairs) dae) are described from the digestive tract of and arrangement of the genital papillae, and by the tail perciform fishes off New Caledonia: H. alatum n. sp. tip which lacks any distinct cuticular projections from Plectropomus laevis (Lace´pe`de) (Serranidae) visible under the light microscope. In addition, and and H. sphyraenae n. sp. from Sphyraena qenie unidentifiable at the species level, conspicuously large Klunzinger (Sphyraenidae). The former species (45.71–66.10 mm long) larvae of Contracaecum (H. alatum) is mainly characterised by its large body Railliet & Henry, 1912, were found in the body cavity (male 42.05 mm, gravid females 51.18–87.38 mm of P. laevis, which serves as a paratenic host for this long), the shape of the dorsal lip, conspicuously broad parasite. cervical alae, a short caecum and a long ventricular appendix, the length of the spicules (925 lm), the number (25 pairs) and distribution of the genital papillae and the tail tip bearing numerous minute Introduction cuticular protuberances. The other species (H. sphyraenae) is mainly characterised by the Our knowledge of the species diversity of anisakid nematodes (Anisakidae) which parasitise marine fishes and elasmobranchs in New Caledonian waters remains incomplete, since only four nominal species of these parasites have previously been recorded in this region: Terranova scoliodontis (Baylis, 1931) F. Moravec (&) from the shark Galeocerdo cuvier (Pe´ron & Lesueur) Institute of Parasitology, Biology Centre of the Czech (Carcharhinidae), Hysterothylacium cenaticum (Bruce Academy of Sciences, Branisˇovska´ 31, & Cannon, 1989) from Tetrapturus audax (Philippi) 370 05 Cˇ eske´ Budeˇjovice, Czech Republic e-mail: [email protected] (Istiophoridae), Raphidascaris (Ichthyascaris) ne- mipteri Moravec & Justine, 2005 from Nemipterus J.-L. Justine furcosus (Valenciennes) (Nemipteridae), and ´ Institut Syste´matique, Evolution, Biodiversite´, ISYEB, R.(Ichthyascaris) etelidis Moravec & Justine, 2012 UMR7205 CNRS, EPHE, MNHN, UPMC, Muse´um National d’Histoire Naturelle, Sorbonne Universite´s, from Etelis coruscans Valenciennes and Pristipo- CP51, 57 rue Cuvier, 75231 Paris cedex 05, France moides filamentosus (Valenciennes) (both Lutjanidae) 123 182 Syst Parasitol (2015) 92:181–195 (Moravec & Justine, 2005, 2006, 2012). Anisakid Family Anisakidae Railliet & Henry, 1912 nematodes unidentified to species or genera, mostly as larval stages, have also been reported from fishes belonging to different families by Justine et al. (2010a, Hysterothylacium alatum n. sp. b, 2012) and Shamsi et al. (2015). Recent examinations of anisakid nematodes col- Type-host: Blacksaddled colargrouper Plectropomus lected from two species of perciform fishes, the laevis (Lace´pe`de) (Perciformes: Serranidae); fork blacksaddled coralgrouper Plectropomus laevis length 550 mm. (Lace´pe`de) (Serranidae) and the blackfin barracuda Site in host: Digestive tract (pyloric caecum). Sphyraena qenie Klunzinger (Sphyraenidae), revealed Type-locality: Off New Caledonia, Passe de Teramba that they represent two new species of Hysterothy- off La Foa (collected 27.iii.2003). lacium Ward & Magath, 1917 plus one species of Prevalence and intensity: 1 of 6 fish infected; 6 larval Contracaecum Railliet & Henry, 1912, which nematode specimens. are described herein. Type-material: Holotype and allotype (anterior and The blacksaddled coralgrouper P. laevis (maximum posterior body ends mounted on SEM stub and body length 125 cm, weight 24.2 kg) and the blackfin remaining parts of bodies preserved in 70 % ethanol) barracuda S. qenie (maximum body length 200 cm, plus 2 paratypes in the Helminthological Collection of weight up to 50 kg) are both tropical marine, reef- the Institute of Parasitology, Biology Centre of the associated fishes. The former has an Indo-Pacific Czech Academy of Sciences, Cˇ eske´ Budeˇjovice (Cat. distribution, whereas the latter is distributed in the No. N–1090); 2 paratypes in the Muse´um National Indo-Pacific from the Red Sea and East Africa to the d’Histoire Naturelle, Paris (MNHN JNC357A). central Indian Ocean, French Polynesia and the eastern Etymology: The specific name alatum (=winged) is a Pacific (Mexico, Panama). Both species are targeted Latin adjective relating to a characteristic feature of by commercial and recreational fishermen (Froese & this nematode, i.e. the presence of conspicuously Pauly, 2015). broad cervical alae. Description (Figs. 1–3) Materials and methods General. Body large, elongate, yellowish in colour. Fish were caught off New Caledonia by various Anterior part of body slender. Broad asymmetrical means. The nematodes intended for morphological cervical alae, widest in region of c. 1/3 of oesophageal studies were fixed in hot 70 % ethanol. For light length, extend posteriorly to about level of middle of microscopical examination (LM), the specimens were oesophagus (Figs. 1D, 2A); from there they continue cleared with glycerine. Drawings were made with the posteriorly to tail as narrow cuticular ridges (Figs. 2E, aid of a Zeiss drawing attachment. Specimens used for F, 3A). Deirids not observed. Anterior end with 3 large scanning electron microscopical examination (SEM) lips provided with broad lateral flanges; each lip has were postfixed in 1 % osmium tetroxide (in phosphate narrow base. Posterior part of dorsal lip roughly buffer), dehydrated through a graded acetone series, rectangular, broad, with margins of lateral flanges critical-point-dried and sputter-coated with gold; they almost parallel, bears 2 lateral double papillae; were examined using a JEOL JSM-7401F scanning anterior part of dorsal lip much narrower, anteriorly electron microscope at an accelerating voltage of 4 kV rounded, separated from posterior part of lip by 2 (GB low mode). All measurements are in micrometres fairly deep lateral depressions (Figs. 1B, 2B–D). unless otherwise indicated. The classification system Subventral lips with 1 double, 1 single papilla and of the Ascaridoidea adopted follows Keys to the amphid. Interlabia well developed, conical, reaching Nematode Parasites of Vertebrates (Anderson et al., anteriorly to about level of labial papillae (Figs. 1C, 2009; Gibbons, 2010). The fish nomenclature follows 2B–D). Oesophagus cylindrical, long, ending in small FishBase (Froese & Pauly, 2015). spherical ventriculus; ventricular appendix narrow, 123 Syst Parasitol (2015) 92:181–195 183 Fig. 1 Hysterothylacium alatum n. sp. ex Plectropomus laevis. A, Anterior end of young female, lateral view; B, Dorsal lip; C, Cephalic end of gravid female, sublateral view; D, Anterior body end of largest female, ventral view; E, Tail of largest female, lateral view; F, Tail tip of largest female, ventral view; G, Tail of male, ventral view; H, Egg; I, Vulva, lateral view; J, Posterior end of male, ventral view; K, Tail of male, lateral view; L, Posterior end of male, lateral view 123 184 Syst Parasitol (2015) 92:181–195 Fig. 2 Hysterothylacium alatum n. sp., SEM micrographs. A, Anterior end of female, ventral view; B–D, Cephalic end, apical, dorsal and subdorsal views, respectively; E, F, Posterior end of male, lateral and ventral views, respectively. Abbreviations: a, double cephalic papilla; b, single cephalic papilla; c, amphid; d, dorsal lip; i, interlabium very long. Anterior intestinal caecum short, slightly maximum width 190. Nerve-ring and excretory pore longer than length of ventriculus (Fig. 1A, D). Cae- 653 and 680, respectively, from anterior extremity. cum to ventricular appendix length ratio 1:8–18. Ventriculus 109 9 136; ventricular appendix Nerve-ring encircles oesophagus at about 1/6 of its 5.30 mm long, maximum width 204. Intestinal cae- length. Excretory pore at level of nerve-ring or just cum 299 long, with maximum width 82. Caecum to posterior to it (Fig. 1A, D). Tail conical; its tip with ventricular appendix length ratio 1:18. Posterior end of numerous minute but distinct protuberances. body curved ventrally. Spicules equal, alate, 925 mm long, representing 2.2 % of body length (Fig. 1L). Male [Based on 1 specimen, holotype.] Length of Total of 25 pairs of genital papillae present, 17 being body 42.05 mm, maximum width 558. Lips 105 long; pre-anals, 1 adanal, and 7 postanals; papillae of several length of interlabia 45. Length of oesophagus posteriormost pre-anal pairs, adanal pair and of 1.90 mm, representing 4.5 % of body length; postanal pairs very small; no double papillae present 123 Syst Parasitol (2015) 92:181–195 185 appendix 4.64–5.85 (4.64) mm long, maximum width 136 (136). Intestinal caecum 320–571 (571) long, maximum width 82–95 (95). Caecum to ventricular appendix length ratio 1:8–18 (1:8). Vulva pre-equato- rial, 26.38–46.44 (46.44) mm from anterior end of body, at 52–53 % (53 %) of body length; posterior vulval lip protrudes distinctly (Fig. 1I). Vagina long (in allotype 3.40 mm long, 163 wide), directed posteriorly from vulva. Uterus forms numerous coils, contains numerous almost spherical eggs 48–51 in diameter in allotype (Fig. 1H); only immature eggs present in paratype. Tail conical, 231–680 long (Fig. 1E); tip rounded, bears numerous small cuticular protuberances (Fig. 1E, F). Non-gravid female [Based on 2 specimens.] Length of body 11.48–14.21 mm, maximum width 258–299. Maximum width of cervical alae 27–41. Lips 75–78 long; length of interlabia 33–36. Length of oesophagus 1.71–1.90 mm, representing 13.4–14.9 % of body length; maximum width 82–109. Nerve-ring and excretory pore 354–367 and 405–408, respectively, from anterior extremity.
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