INTRODUCTION
Marine nematodes are one of the most abundant animal groups, discovered in various marine habitats, from beach sediment to deep-sea environments (Nicholas 1984). In addition, many species have been reported from marine algal habitat as well as on the surface of other invertebrates (Porifera, Cnidaria, Bryozoa and Mollusca and so on) (Kito and Hope 1999). Free-living marine nematodes are used as environmental bio-indicator in marine benthic ecosystems because they show high biomass, species richness, and shorter generation period (Heip et al. 1985;Sandulli and De Nicola-Giudici 1991;Schratzberger et al. 2000;Balsamo et al. 2010). They also play an important role in the marine food chain because they are predators that feed on other invertebrate such as unicellular algae, bacteria, other nematodes, and further prey on fish and other large invertebrates (Bouwman 1983;Heip et al. 1985;Giere 2008).
The first taxonomic study of free-living marine nematodes in Korea was began with the description of Tenuidraconema koreensis by Rho and Kim (2004). Since then, 70 species of free-living marine nematodes have been recorded by various taxonomists (Rho and Kim 2004, 2005;Lim and Chang 2006;Rho et al. 2010;Rho and Min 2011;Barnes et al. 2012;Hong et al. 2016;Jeong et al. 2019a, b, 2020;Lee and Rho 2019;Rho et al. 2020;Tchesunov et al. 2020, 2021, 2022;Lee et al. 2021a, b, 2022). Among them, 33 species were reported in the East Sea, 8 in the southern coast, 5 in the Yellow Sea, and 24 in Jejudo Island of Korea.
As a result of continuous biodiversity study of free-living marine nematodes around Korea, three nematode species were discovered from shallow intertidal and subtidal sediments in the benthic environment. Present paper deals with morphological study of three unrecorded free-living marine nematodes using a differential interference contrast (DIC) microscope.
MATERIALS AND METHODS
Specimen collecting. Specimens were collected from upper surface of the intertidal sandy beach of southern and western coasts of Korea using a hand scoop, and also obtained from shallow subtidal zone around the East Sea of Korea using Smith-McIntyre grab. Meiobenthic animals were roughly separated from the sediment the freshwater shock technique, and then quickly filtered through a 63 μm mesh-sieve and fixed in 5% buffered formalin in seawater (Kristensen and Higgins 1989). Afterwards, the meiobenthos was extracted by flotation in Ludox (DuPont, USA) HS 40 in three centrifugation cycles (Burgess 2001).
Morphological observation. Nematodes were picked out from the mixed meiobenthos under high magnification of dissecting microscope (LEICA 205C; Wetzlar, Germany). Target nematodes were transferred to 5% glycerin solution, and then dehydrated through a graded series of glycerine. Nematodes were mounted on between two coverslips using the HS slide (Shirayama et al. 1993). Mounted nematodes were measured and examined using an Olympus BX 53 microscope equipped with Nomarski differential interference contrast and photographed with an Olympus DP 26 digital camera. Measurements were carried out with an Olympus Cellsens corresponding imaging software (Olympus, Tokyo, Japan). Line drawings for the mounted specimens were developed by an Olympus BX 53 microscope with drawing tube, and were adjusted in Adobe illustrator CC program. All the measurements are given in μm.
SYSTEMATIC ACCOUNTS
Phylum Nematoda Potts, 1932
Class Chromadorea Inglis, 1983
Order Demoscolecida Filipjev, 1929
Family Desmoscolecidae Shipley, 1896
Genus Desmoscolex Claparède, 1863
Subgenus Desmoscolex Claparède, 1863
Desmoscolex (Desmoscolex) maxTimm, 1970 (Figs. 1, 2)
Desmoscolex (Desmoscolex) maxTimm, 1970, p. 26, figs. 26-28.
Desmoscolex max: Timm, 1978, p. 229, fig. 2E, F;Decraemer, 1984, p. 311, fig. 3C, D.
Material examined. The specimen was collected from the subtidal zone of Wangdolcho, Hupo-myeon, Uljin-gun, Gyeongsangbuk-do, Korea, 26 Sep. 2019 (leg. H Lee). One female (MABIK NA00157826), in glycerin on a HS slide, was deposited in the nematode collection at the specimen conservation room of the Marine Biodiversity Institute of Korea (MABIK), Seocheon, Korea.
Habitat. Subtidal coarse sediment, 68 m depth (36°42ʹ 26.52″N, 129°43′07.22″E).
Measurements. Female (n=1): L=419; hd=20; cs=34; sd1=28; sd3=30; sd5=29; sd7=27; sd9=25; sd11=26; sd13=36; sd16=40; sv1=19; sv2=18; sv4=19; sv6=17; sv8=20; sv12=16; sv14=14; sv15=18; sv17=20; ph=82; t=89; tmr=46; t/tmr=1.9; tmrw=20; tmr/ tmrw=2.3; mbd=67; mbde=42; a=6.3; b=5.1; c=4.7.
Description. Female. Body small, ventrally curved, slightly tapered anteriorly and posteriorly; tail end ring strongly curved dorsally. Body cuticle with 17 main rings of slightly different widths and lengths; interzone very wide with uncovered annules. Each main ring covered with layer of secretion and different sized foreign material (Figs. 1A, 2A).
Somatic setae arranged according to untypical desmoscolecid setal pattern: eight pairs of subdorsal somatic setae on main rings 1, 3, 5, 7, 9, 11, 13, 16; nine pairs of subventral somatic setae on main rings 1, 2, 4, 6, 8, 12, 14, 15, 17. Subdorsal somatic setae gradually becoming finer to tip, without mid-central canal, longer than subventral somatic setae, and not inserted on peduncle. Pairs of subdorsal somatic setae on main rings 13 and 16 longer than rest of subdorsal somatic setae. Subventral somatic setae smaller than subdorsal somatic setae and relatively slender, not inserted on peduncle; cylindrical basal part broad, without mid-central canal, tapered to tip. Subventral somatic setae all about same length, except for those on main rings 15 and 17, which are slightly longer (Fig. 1A-C).
Head triangle-shaped, obviously elongated, longer than wide, anteriorly very tapered (Figs. 1A, 2A). Cephalic cuticle slightly covered with thin layer of fine granular desmos, except in amphideal zone. Long hairy cephalic setae situated at almost middle of head, 34 μm long; not jointed and not inserted on peduncles. Amphideal fovea very long (34 μm) and elongated, largely covering head laterally, reached at anterior margin of main ring 1.
Digestive system typical of Desmoscolex. Buccal opening with minute buccal cavity. Oesophagus 82 μm long, posteriorly extending to level of anterior margin of main ring 4 (Fig. 1B). A pair of ocelli situated at level of posterior margin of main ring 4. Vulva not observed. Annus situated at between main rings 15 and 16.
Tail with two main rings (Figs. 1C, 2C). End ring 46 μm long, about 2.3 times longer than wide, consisting of conical anterior part (32 μm long) and well-developed naked spinneret (14 μm long). Terminal somatic setae situated at level of anterior margin of end ring subventrally. End ring, with exception of terminal spinneret, totally covered with desmos. Caudal glands not observed.
Remarks.Desmoscolex (D.) maxTimm, 1970 belongs to the species group with 17 main rings and the untypical setal pattern. Desmoscolex (D.) max is mainly distinguished from its congeners in having the following characteristics: (1) the presence of untypical setae arrangement (eight subdorsal somatic setae and nine subventral somatic setae), (2) the presence of obviously elongated triangle-shaped head, (3) the presence of long hairy cephalic setae, (4) the absence of peduncle at the base of somatic setae, and (5) end ring has a long naked spinneret. The present Korean specimen of D. (D.) max agrees well with Timm’s (1970) original description, especially in having the elongated, triangle shaped head and a long naked spinneret in the end ring. However, the Korean specimens of D. (D.) max are not slightly accorded with Timm’s (1970) original description in the number of subventral somatic setae as follows: compared to 10 of the original description, there are 9 of Korean specimen, and main ring 10 do not have subventral somatic setae in the Korean specimen.
Distribution. Galapagos Islands (Timm 1970, 1978), Moçambique Channel (Decraemer, 1984) and Korea (Present study).
Order Monhysterida Filipjev, 1929
Family Xyalidae Chitwood, 1951
Genus Daptonema Cobb, 1920
Daptonema longiapophysisHuang and Zhang, 2010 (Figs. 3, 4; Table 1)
Daptonema longiapophysisHuang and Zhang, 2010, p. 391-394, figs. 1, 2.
Material examined. Specimens were collected from Ttangkkeutsongho Beach, Songji-myeon, Haenam-gun, Jeollanam-do, Korea, 27 May 2020 (leg. HS Rho and H Lee). One male (MABIK NA00157825), in glycerin on a HS slide, was deposited in the nematode collection at the specimen conservation room of the Marine Biodiversity Institute of Korea (MABIK), Seocheon, Korea. Three males (KIOST NEM-1-2673, KIOST NEM-1-2674, KIOST NEM-1-2675) in glycerin on HS slide, were deposited in the nematode collection at the specimen conservation room of the Bio-Resources Bank of Marine Nematodes (BRBMN), East Sea Research Institute, Korea Institute of Ocean Science & Technology (KIOST), Korea.
Habitat. Intertidal fine sand beach (34°18′54.00″N, 126° 31′06.00″E).
Measurements. See Table 1 for detailed measurements and morphometric ratios.
Description. Males. Body cylindrical and slender (1,421- 1,565 μm long), gradually tapering towards both extremities (Fig. 4A). Cuticle with coarse annulations, which begins in buccal cavity and extends to tail tip (Fig. 4C). Maximum body diameter at mid body level 39-44 μm wide. Head end spherical (18-21 μm wide), with six large and raised lips, each with six setiform labial sensilla. Six relatively short outer labial setae (11-15 μm long) and six longer cephalic setae (16-18 μm long) arranged in one circle (Fig. 3C). Buccal cavity spacious and conical (11-14 μm wide) without teeth (Figs. 3B, 4B). Somatic setae thin filamentous, scattered throughout body. Amphideal fovea not visible. Pharynx cylindrical, 357-391 μm long, about 24-26% of total body length. Pharyngo-intestinal junction with cardia. Nerve ring encircling pharynx, situated at 31- 34% of pharynx length from anterior end (Fig. 3A). Tail with anteriorly conical, tapering with distal third cylindrical, 181-195 μm long. Tail tip bearing two terminal setae. Three caudal glands with common opening (Figs. 3E, 4F).
Spicules paired, slightly curved, and 23-28 μm long arc, 21-25 μm long chord. Both spicules’ proximal end with capitulum, and distal end with bifid tip. Spicules with a projection in middle part each of dorsal and ventral side. Gubernaculum loop shaped (25-26 μm long), parallel to spicule, with long dorso-caudal apophysis (Figs. 3D, 4D, E). One pre-cloacal and one post-cloacal setae present.
Remarks.Datonema longiapophysis was firstly described from intertidal sandy sediments on the Rizhao coast of Shandong province and the Huangdao coast of Qingdao from the Yellow Sea, China. Daptonema longiapophysis is characterized by the presence of setiform labial sensilla, spicules with a projection on both sides, and gubernaculum with dorso-caudal apophysis longer than spicules length.
The present Korean specimens of D. longiapophysis are very similar to D. deconinckiSharma, 1985 in having the spicules with bifid distal tip and gubernaculum with long dorso-caudal apophysis (Sharma 1985). However, D. longi- apophysis can be easily distinguished from D. deconincki by the relatively longer body length (1,421-1,565 μm vs. 942- 1,157 μm in males), shorter spicules length (23-28 μm vs. 45-51 μm), and lower ratio of gubernaculum apophysis to spicules length (0.9-1.1 vs. 1.8). The present Korean specimens of D. longiapophysis are most like the original description by Huang and Zhang (2010), especially in the body size and structure, the shape of head and tail, the presence of spicules with a projection in the middle part, and the presence of long dorsal-caudal apophysis. However, the Korean specimens of D. longiapophysis are not accorded with the original description by having a bifid disal tip in the spicules.
Distribution. China (Huang and Zhang 2010) and Korea (Present study).
Order Monhysterida Filipjev, 1929
Family Xyalidae Chitwood, 1951
Genus PseudosteineriaWieser, 1956
Pseudosteineria sinicaHuang and Li, 2010 (Figs. 5, 6; Table 2)
Pseudosteineria sinicaHuang and Li, 2010, p. 2454-2458, figs. 1, 2.
Material examined. Specimens were collected from Gamami Beach, Hongnong-eup, Yeonggwang-gun, Jeollanam- do, Korea, 19 Jun 2018 (leg. HG Kim and HJ Lee). One male (MABIK NA00157824), in glycerin on a HS slide, was deposited in the nematode collection at the specimen conservation room of the Marine Biodiversity Institute of Korea (MABIK), Seocheon, Korea. Two males (KIOST NEM-1-2676, KIOST NEM-1-2677) in glycerin on HS slide, were deposited in the nematode collection at the specimen conservation room of the Bio-Resources Bank of Marine Nematodes (BRBMN), East Sea Research Institute, Korea Institute of Ocean Science & Technology (KIOST), Korea.
Habitat. Intertidal mud-sand beach (35°23′55.90″N, 126° 24′17.85″E).
Measurements. See Table 2 for detailed measurements and morphometric ratios.
Description. Males. Body slender, 1,297-1,580 μm long, gradually tapering towards both extremities (Fig. 6A). Cuticle transversely annulated, beginning at base of buccal cavity and extends to tail tip (Fig. 6C). Maximum body diameter at mid body level, 59-62 μm long. Head end slightly inflated with six lips. Labial region with six inner labial sensilla papilliform and 12 outer labial sensillae setiform arranged in two circles. Six longer outer labial setae 7-9 μm long, six shorter cephalic setae 5-6 μm long. (Fig. 5C). Buccal cavity funnel-shaped, without teeth. Subcephalic setae arranged in eight longitudinal rows on both sides of body, and located at directly behind cephalic setae. Each group with three to five subcephalic setae, increasing gradually from anterior to posterior setae in every row. Length of shortest subcephalic setae 7-12 μm long and longest one 43-51 μm long (Figs. 5B, 6B). Somatic setae scattered throughout body. Amphideal fovea not visible. Pharynx cylindrical, 249-315 μm long, about 19-20% of total body length. Pharyngo-intestinal junction with cardia. Nerve ring encircling pharynx, situated at 36-40% of pharynx length from anterior end (Fig. 5A). Tail conico-cylindrical, 163-194 μm long with distal fourth cylindrical part. Tail tip bearing three to four terminal setae; longest setae up to 32-42 μm long (Figs. 5E, 6F). Three caudal glands observed.
Spicules paired, slightly curved, and unequal in length. Left spicule longer (57-62 μm long as arc) than right spicule (47-52 μm long as arc), and left spicule with a slight contraction in middle part. Both spicules’ proximal end with large capitulum and distal taper tip. Gubernaculum arcuated, tapered distal part, with dorso-caudal apophysis (Figs. 5D, 6D, E). Precloacal supplements absent.
Remarks. The genus Pseudosteineria was first erected by Wieser (1956), and currently 16 valid species have been reported from all over the world (Wieser 1956;Tchesunov 2000;Huang and Li 2010;Sun et al. 2019;Cidreira et al. 2020). Pseudosteineria sinicaHuang and Li, 2010 was firstly described from intertidal sandy sediment at Rizhao coast of the Yellow Sea, China. Pseudosteineria sinica is characterized by unobservable amphideal fovea, different length of spicules: left spicule divided into two sections jointed in the middle and longer than right spicule, both spicules with proximal capitulum, tapered distally, and gubernaculum with dorso-caudal apophysis.
The present Korean specimens of P. sinica are very similar to P. inaequispiculata (Platonova 1971) by unequal spicules, but P. inaequispiculata are clearly distinguished by having round amphideal fovea and the absence of gubernaculum apophysis (Platonova 1971). The present Korean specimens are most like the original description by Huang and Li (2010) in the following features, such as the absence of amphideal fovea, the presence of different length of spicules, and gubernaculum with dorso-caudal apophysis. However, the Korean specimens of P. sinica are not slightly accorded with the original description by the number of subcephalic setae (three to five vs. three to four), and shorter length of the shortest subcephalic setae (8-12 μm vs. 15-18 μm).
Distribution. China (Huang and Li 2010) and Korea (Present study).