Morphological and molecular studies of the life-cycle stages of the monorchiid Monorchis parvus (Looss, 1902) (Digenea) from the Southern Mediterranean coast (Tunisia).

The elucidation of life-cycles of digeneans, with their successive larval stages, is facilitated by the use of molecular markers. Samples of sporocysts containing cercariae and metacercariae belonging to Monorchis Monticelli, 1893 were collected from naturally infected bivalves, Cerastoderma glaucum (Bruguière, 1789), and adult forms of Monorchis spp. were collected from sparid fishes of the genus Diplodus. All specimens were collected in the Gulf of Gabès, southern Tunisia. The identities of the examined molluscs and fishes were determined via molecular barcoding of their COI gene. Sequences of COI and ITS1 genes were also obtained for both larval and adult stages of collected parasite specimens. Genetic sequence data generated for the collected larval specimens only differed minimally from the sequence data of adults identified as Monorchis parvus; we attribute the difference to intraspecific variation. The morpho-anatomical study showed that the different stages of M. parvus collected from the Tunisian coasts had the same morphology as those reported in European waters with a lag in maturity and lower measurements. The species is recorded and molecularly characterised for the first time off the Tunisian coasts.

A world of taxonomic pain: cryptic species, inexplicable host-specificity, and host-induced morphological variation among species of Bivesicula Yamaguti, 1934 (Trematoda: Bivesiculidae) from Indo-Pacific Holocentridae, Muraenidae and Serranidae.

The taxonomy of species of Bivesicula Yamaguti, 1934 is analysed for samples from holocentrid, muraenid and serranid fishes from Japan, Ningaloo Reef (Western Australia), the Great Barrier Reef (Queensland), New Caledonia and French Polynesia. Analysis of three genetic markers (cox1 mtDNA, ITS2 and 28S rDNA) identifies three strongly supported clades of species and suggests that Bivesicula as presently recognized is not monophyletic. On the basis of combined morphological, molecular and biological data, 10 species are distinguished of which five are proposed as new. Bivesicula Clade 1 comprises seven species of which three are effectively morphologically cryptic relative to each other; all seven infect serranids and four also infect holocentrids. Bivesicula Clade 2 comprises three species of which two are effectively morphologically cryptic relative to each other; all three infect serranids and one also infects a muraenid. Bivesicula Clade 3 comprises two known species from apogonids and a pomacentrid, and forms a clade with species of Paucivitellosus Coil, Reid & Kuntz, 1965 to the exclusion of other Bivesicula species. Taxonomy in this genus is made challenging by the combination of low resolving power of ribosomal markers, the existence of regional cox1 mtDNA populations, exceptional and unpredictable host-specificity and geographical distribution, and significant host-induced morphological variation.

Redescription and molecular characterization of two species of Pauciconfibula (Monogenea, Microcotylidae) from trachinid fishes in the Mediterranean Sea.

Many Pauciconfibula spp. have a long and complicated taxonomic history. The remaining unsolved taxonomic confusion in this genus is impelled by the host range and status of Pauciconfibula spp. from trachinid fishes: Pauciconfibula trachini and Pauciconfibula draconis, from Trachinus radiatus and Trachinus draco (Trachinidae), respectively. Pauciconfibula trachini was reported on Trachinus draco, type host of Pauciconfibula draconis suggesting thus a stenoxenic specificity for the former monogenean and the occurrence of two congeneric polyopisthocotyleans on a single host. Moreover, the validity of Pauciconfibula draconis was repeatedly questioned by several authors, unjustified synonymy between the two species was proposed, and the delimitations between the two species remained unsolved. Original descriptions were also incomplete and poorly illustrated. In this study, we provide a detailed illustrated redescription of both species based on newly collected specimens of Pauciconfibula trachini and Pauciconfibula draconis collected from their type hosts from off three Mediterranean localities: Algeria, Tunisia, and Italy. Integrative taxonomy using COI sequences was applied to resolve the delimitation between Pauciconfibula trachini and P. draconis. This study provides the first DNA barcoding for members of this genus.

Some adult and larval nematodes from fishes off New Caledonia.

A collection of nematodes from the Muséum National d'Histoire Naturelle (National Museum of Natural History), Paris, France, was studied. The nematodes had previously been collected from a range of marine fish hosts. The aim of this study was to investigate the identity of these nematodes. Detailed body measurements of the nematodes were taken via light microscopy and where possible first and second internal transcribed spacers (ITS-1 and ITS-2) of ribosomal DNA were subjected to PCR, purified and sequenced. Six species and three larval types were found from a range of fish species and included 13 new host records and two new geographical records. Of the taxa found, Hysterothylacium kajikiae is the only species which may have zoonotic potential. The majority of host fish in this study are edible species, commercially and recreationally fished and frequently incorporated into the cuisine of New Caledonia as raw fish dishes. This study will allow better understanding of the diversity, life cycles, distribution and host-parasite relationships in the New Caledonia area.

Paracapillaria (Paracapillaria) gastrica n. sp. (Nematoda: Capillariidae) from the marine fish Synodus variegatus Lacépède (Synodontidae, Aulopiformes) off New Caledonia.

Based on light and scanning electron microscopical studies, a new nematode parasite, Paracapillaria (Paracapillaria) gastrica n. sp. (Capillariidae), is described from the stomach of the marine fish (variegated lizardfish) Synodus variegatus (Lacépède) (Aulopiformes: Synodontidae) from off New Caledonia. This species is mainly characterised by the length of the spicule (267 µm), the proximal spicule end expanded to form a conspicuous folded, lobular rim, the presence of a well-developed caudal bursa supported by two lateral projections (rays) not adhering to the posterior border of body and by the size of eggs (54-60 × 24-27 µm) with non-protruding polar plugs. Paracapillaria gastrica n. sp. is the first known capillariid species parasitic in a host belonging to the fish order Aulopiformes and the first species of this genus reported from fishes in New Caledonian waters and the South Pacific Ocean.

Redescription of Flexophora ophidii Prost & Euzet, 1962 (Monogenea: Diclidophoridae) from Ophidion barbatum (Ophidiidae) off the Algerian coast, Mediterranean Sea.

The diclidophorid Flexophora ophidii Prost & Euzet, 1962, the type- and only species of the genus Flexophora Prost & Euzet, 1962, has never been redescribed neither reported since its original description. The latest lacked a diagnosis of the genus that remains unavailable to date. Flexophora ophidii is redescribed and illustrated based on examination of new material from the type-host Ophidion barbatum L. (Ophidiiformes: Ophidiidae) collected off the Algerian coast (new geographical record), southern Mediterranean. A diagnosis of the genus is provided.

Towards the resolution of the Microcotyle erythrini species complex: description of Microcotyle isyebi n. sp. (Monogenea, Microcotylidae) from Boops boops (Teleostei, Sparidae) off the Algerian coast.

The monogenean Microcotyle erythrini is atypical because it has been recorded from several fish host species in the Mediterranean Sea and Atlantic Ocean, in contrast to many species which are considered strictly specific. This could indicate a true lack of specificity or that several cryptic species are involved. This paper is a partial attempt to solve this problem. Specimens of a monogenean resembling M. erythrini were collected from bogues, Boops boops, caught off Algeria. A comparison with published descriptions and with museum specimens of M. erythrini did not yield any clear morphological difference. However, sequences of cytochrome c oxidase subunit I (COI) differed by 16.3% from that of M. erythrini (from GenBank, material collected from the type-host Pagellus erythrinus), indicating that the species was different. The species from B. boops is therefore described here as Microcotyle isyebi n. sp. and differential diagnoses with Microcotyle species from the Mediterranean and from sparids are provided. These results suggest that a molecular re-evaluation of other M. erythrini-like specimens from various fish hosts could reveal the existence of additional parasite biodiversity.

Dichelyne (Cucullanellus) bodiani n. sp. and Cucullanus hansoni Olsen, 1952 (Nematoda: Cucullanidae) from fishes off New Caledonia.

Two species of cucullanid nematodes collected from the intestine of marine fishes off New Caledonia were studied with the use of light and scanning electron microscopy (SEM): Dichelyne (Cucullanellus) bodiani n. sp. from Bodianus perditio (Quoy & Gaimard) (Perciformes: Labridae), characterised mainly by the small size of the body (males and gravid females 2.26-3.13 mm and 2.46-3.32 mm long, respectively), the presence of very long spicules (1.53-1.66 mm in length), the remarkably large seminal vesicle and the arrangement of caudal papillae, is the second known species of Dichelyne Jägerskiöld, 1902 parasitising fishes of the Labridae and the second nominal species of this genus recorded from fishes in New Caledonian waters. Cucullanus hansoni Olsen, 1952, originally described from Hawaii, is now, after 67 years, again reported from its type-host, Sufflamen fraenatum (Latreille) (Tetraodontiformes: Balistidae), from off New Caledonia. The SEM examination of C. hansoni, used in this species for the first time, revealed some new morphological details, such as the presence of a median precloacal cuticular elevation or the shape of deirids and distal tips of spicules, as well as the exact location of caudal papillae and phasmids in the male. This is the seventh species of Cucullanus Müller, 1777 recorded from fishes off New Caledonia.

Redescription and molecular characterisation of Allogastrocotyle bivaginalis Nasir & Fuentes Zambrano, 1983 (Monogenea: Gastrocotylidae) from Trachurus picturatus (Bowdich) (Perciformes: Carangidae) off the Algerian coast, Mediterranean Sea.

Allogastrocotyle bivaginalis Nasir & Fuentes Zambrano, 1983, the sole species of Allogastrocotyle Nasir & Fuentes Zambrano, 1983, was described from Trachurus lathami Nichols off Venezuela and never recorded since. We found monogeneans on Trachurus picturatus (Bowdich) (Carangidae) off the Algerian coast, Mediterranean Sea, which had the characteristics of the genus, including, especially, paired vaginae. We compared them to the single available specimen of A. bivaginalis, the holotype. Unfortunately, this holotype does not show clamp structure and soft internal anatomy. Our specimens were similar to A. bivaginalis in most characteristics such as body shape, vaginal openings, and number of testes, clamps and hooks in genital atrium. We detected minor differences in clamp structure but could not ascertain if these were the result of incomplete observations in the original description of real morphological differences. The host fish of our specimens were barcoded (cox1) confirming their specific identity. A phylogenetic analysis of cox1 sequences showed that our sequences of A. bivaginalis were distinct from those of Pseudaxine trachuri Parona & Perugia, 1889 (distance > 15%) and of several other gastrocotylids. Finally, we could not distinguish our Mediterranean specimens from A. bivaginalis, neither on the base of morphology (because the original description is incomplete) nor on molecules (because molecular information is lacking on A. bivaginalis from Venezuela) and ascribe them to A. bivaginalis. However, hosts are different, and localities are widely separated, so it is likely that future study will show that the species from the Mediterranean is a distinct, new, species.

Microcotyle visa n. sp. (Monogenea: Microcotylidae), a gill parasite of Pagrus caeruleostictus (Valenciennes) (Teleostei: Sparidae) off the Algerian coast, Western Mediterranean.

Parasite biodiversity of fish of the southern part of the Mediterranean sea is still incompletely explored. We describe here Microcotyle visa n. sp. from the gill filaments of the bluespotted seabream Pagrus caeruleostictus (Valenciennes) (Sparidae) collected off the Algerian coast. The identity of fish hosts was confirmed by barcoding. Microcotyle visa n. sp. is herein described and illustrated. Analysis of the cox1 gene of the monogeneans revealed minor intraspecific variation (1.4%), an order of magnitude lower than the distance between this species and other Microcotyle species (10-15 %). Microcotyle visa n. sp. is distinguished from Microcotyle erythrini van Beneden & Hesse, 1863, a congener infesting sparids, on the basis of morphological (size of clamps, number of testes) and molecular (cox1) differences. This is the fourth member of the genus known to parasitise a sparid host. A species of Paramicrocotyle sp. included in the molecular analysis was nested within a robust Microcotyle + Paramicrocotyle clade; in the absence of demonstrated molecular and morphological differences, we consider that Paramicrocotyle Caballero & Bravo-Hollis, 1972 is a junior synonym of Microcotyle van Beneden & Hesse, 1863 and transfer two species of Paramicrocotyle as Microcotyle danielcarrioni (Martinez & Barrantes, 1977) n. comb. and Microcotyle moyanoi (Villalba & Fernandes, 1986) n. comb.

Occurrence of anisakid parasites in marine fishes and whales off New Caledonia.

Anisakid nematodes are the most infamous parasites occurring in seafood with ability to infect humans. In the present study, the infective stages of five anisakid larval types, including Anisakis types I and III, Terranova types I and II and Contracaecum larval type, as well as adult Anisakis paggiae are reported from 16 host species from New Caledonian waters. The specific identity of the larval types was investigated using ITS sequence data. Anisakis larval types I and III were identified as Anisakis typica and Anisakis brevispiculata, respectively, based on identical ITS sequences. However, the specific identity of the Terranova larval types and Contracaecum larval type remains unknown until a matching ITS sequence from a well-identified adult is available. Several fish host species are reported for the first time for anisakid larval types found in this study. Considering that third-stage larvae of anisakids are known to be the infective stage of the parasite for humans and the popularity of seafood in New Caledonia, presence of these parasites in New Caledonian fish is of high importance in terms of public health and raising awareness among various stakeholders. Although adult nematodes in the present study were identified as Anisakis paggiae, the spicule length is shorter in our specimens and falls within the range reported for Anisakis oceanicus previously reported in Pacific waters from black fish (genus Globicephala) and later synonymised with Anisakis physeteris. However, our specimens are different from A. physeteris in morphology of ventriculus. Anisakis paggiae has been reported from whales in southern hemisphere and this is the first report from the Pacific regions.

Two new species of nematode parasites, Cucullanus epinepheli sp. n. (Cucullanidae) and Procamallanus (Spirocamallanus) sinespinis sp. n. (Camallanidae), from marine serranid and haemulid fishes off New Caledonia.

Based on light and scanning electron microscopical studies, two new species of parasitic nematodes are described from marine perciform fishes off New Caledonia: Cucullanus epinepheli sp. n. (Cucullanidae) from the intestine of the brownspotted grouper Epinephelus chlorostigma (Valenciennes) (Serranidae) and Procamallanus (Spirocamallanus) sinespinis sp. n. from the intestine of the silver grunt Pomadasys argenteus (Forsskål) (Haemulidae). Cucullanus epinepheli sp. n. differs from its congeners mainly in possessing a unique structure of the anterior, elevated cloacal lip with a large posterior outgrowth covering the cloacal aperture and in the presence of cervical alae and two small preanal papillae on the median dome-shaped precloacal elevation. This is the second known nominal species of this genus parasitising fishes of the family Serranidae and the second representative of Cucullanus Müller, 1777 recorded from fishes in New Caledonian waters. Procamallanus (Spirocamallanus) sinespinis sp. n. is mainly characterised by 10-12 spiral ridges in the buccal capsule, the presence of wide caudal alae, three pairs of pedunculate preanal papillae, two unequally long spicules (465-525 µm and 218-231 µm) and by the tail tip with a knob-like structure in the male, and the broad, rounded tail with a terminal digit-like protrusion without cuticular spikes in the female. This is the fifth nominal species of the subgenus Spirocamallanus Olsen, 1952 reported from fishes in New Caledonian waters.

Redescription of Philometra filiformis (Stossich, 1896) (Nematoda: Philometridae), a gonad-infecting parasite of the common pandora Pagellus erythrinus (Linnaeus) (Sparidae) in the Mediterranean Sea, including new taxonomic features revealed by SEM.

The insufficiently known nematode species Philometra filiformis (Stossich, 1896) (Philometridae) is redescribed based on light and scanning electron microscopical (SEM) examinations of specimens collected from the ovaries of the type-host, the common pandora Pagellus erythrinus (Linnaeus) (Perciformes, Sparidae), from off the Mediterranean coast of Tunisia. The male of this species was studied with SEM for the first time, which revealed some new, taxonomically important morphological features. The male posterior end has a caudal mound consisting of two lateral parts widely separated dorsally from each other, four pairs of small adanal papillae, a pair of large but moderately developed papillae located posterior to the cloacal aperture and a pair of small phasmids. The distal end of the gubernaculum is unique among all but one gonad-infecting species of Philometra Costa, 1845 by the chevron-shaped dorsal lamellar structures forming a median longitudinal range. Philometra filiformis is for the first time reported from Tunisian coastal waters, being the seventh philometrid species so far recorded from marine fishes off Tunisia.

Hamacreadium cribbi n. sp. (Digenea: Opecoelidae) from Lethrinus miniatus (Forster) (Perciformes: Lethrinidae) from New Caledonian waters.

A new species of Hamacreadium Linton, 1910, H. cribbi n. sp. is described from Lethrinus miniatus (Forster) from the waters off New Caledonia. It is compared with the other species of Hamacreadium reported from lethrinids and is characterised by the size of its eggs which tend to be larger [72-93 (84) vs 54-81 (56) µm long] than those of other species. Other characteristics, such as body size and shape and internal ratios, differentiate H. cribbi from other species; these differences are discussed in detail.

Descriptions of Philometra aenei n. sp. and P. tunisiensis n. sp. (Nematoda: Philometridae) from Epinephelus spp. off Tunisia confirm a high degree of host specificity of gonad-infecting species of Philometra Costa, 1845 in groupers (Serranidae).

Based on light and electron microscopical studies of males and mature females, two new gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) are described from the ovary of groupers, Epinephelus spp. (Perciformes; Serranidae), in the Mediterranean Sea off Tunisia (near Sfax): Philometra aenei n. sp. from the white grouper E. aeneus (Geoffroy Saint-Hilaire) and P. tunisiensis n. sp. from the goldblotch grouper E. costae (Steindachner). Identification of both fish hosts was confirmed by barcoding of the infected fish specimens. Philometra aenei is mainly characterised by the length of conspicuously distended spicules (108-123 µm), the presence of a distinct dorsal barb at the middle region of the gubernaculum and a distinct protuberance consisting of two dorsolateral lamellar parts separated from each other by a smooth median field at its distal tip, a V-shaped mound on the male caudal extremity and by the body length of the males (2.34-3.05 mm). The male of this species was found to possess minute deirids in the cervical region, which is quite exceptional within the Philometridae. Philometra tunisiensis is distinguished from other gonad-infecting congeneric species parasitising serranids by the length of the needle-like spicules and gubernaculum (201-219 and 78-87 µm, respectively), spicule length representing 9-11% of body length, the gubernaculum/spicules length ratio of 1:2.52-2.77, the length of oesophagus in the male comprising 15-16% of the body length, the absence of a dorsal protuberance on the distal lamellar part of the gubernaculum and a pair of large papillae posterior to the cloaca, a dorsally interrupted mound on the male caudal extremity and the body length of the male (2.01-2.42 mm). The presence of three morphologically very different species of Philometra in congeneric hosts in the Mediterranean Sea confirms a high degree of host specificity of these gonad-infecting nematodes parasitising groupers.

Neomultitestis aspidogastriformis Bray and Cribb, 2003 (Digenea, Lepocreadiidae): mature spermatozoon and sperm morphologies in the Lepocreadioidea.

Spermatological characteristics are described for the first time in the genus Neomultitestis Machida, 1982, belonging to the family Lepocreadiidae Odhner, 1905. Adult Neomultitestis aspidogastriformis were collected from the digestive tract of Platax teira, caught off New Caledonia. The mature spermatozoon of N. aspidogastriformis is described by transmission electron microscopy (TEM). It includes two axonemes with 9 + "1" pattern of Trepaxonemata, an antero-lateral electron-dense material, attachment zones, two mitochondria, a nucleus, two bundles of parallel cortical microtubules, external ornamentation of the plasma membrane, spine-like bodies, and granules of glycogen. Certain ultrastructural characteristics of the spermatozoon, especially the antero-lateral electron-dense material, are shared by N. aspidogastriformis and all species of the superfamily Lepocreadioidea studied so far. The antero-lateral electron-dense material, present in different families of lepocreadioids, supports the monophyletic status of the superfamily. We also list several ultrastructural characteristics which characterize the mature spermatozoon of the Lepocreadioidea.

New records of species of Philometra (Nematoda: Philometridae) from marine fishes off New Caledonia, including P. cephalopholidis sp. n. from Cephalopholis sonnerati (Serranidae).

Two different species of Philometra Costa, 1845 were collected from marine perciform fishes, the tomato hind Cephalopholis sonnerati (Valenciennes) (Serranidae) and the painted sweetlips Diagramma pictum (Thunberg) (Haemulidae), from off New Caledonia, South Pacific. Nematodes (only males) from the gonad of C. sonnerati represent a new taxon, P. cephalopholidis sp. n., which is mainly characterized by almost equally long spicules (length 186-228 µm), the shape and structure of the gubernaculum with a dorsally lamellate distal tip, and the structure of the caudal end. The nematodes (only gravid females) from abdominal tissues of D. pictum may represent an undescribed species, but, because of the absence of conspecific males, they could not be specifically identified. Philometra cephalopholidis is the sixth nominal species of this genus recorded from fishes off New Caledonia and the thirteenth species of these parasites described from fishes of the family Serranidae.

Ultrastructural characters of the spermatozoa in Digeneans of the genus Bianium Stunkard, 1930 (Digenea, Lepocreadiidae) parasites of fishes: a comparative study of Bianium plicitum and Bianium arabicum.

We present in this paper the first ultrastructural study of the spermatozoon of species of the genus Bianium Stunkard, 1930, namely Bianium plicitum and Bianium arabicum belonging to the family Lepocreadiidae. Specimens are parasites of the tetraodontid teleosts Lagocephalus laevigatus caught in the Atlantic Ocean and Lagocephalus sceleratus caught in Pacific Ocean, respectively. Live worms were fixed and processed for transmission electron microscopy. Observations of numerous cross and longitudinal sections of the spermatozoa of B. plicitum and B. arabicum demonstrated the general pattern described in all previously studied Lepocreadioidea, namely two axonemes with 9+ "1" pattern of Trepaxonemata, two mitochondria, a nucleus, parallel cortical microtubules, and an external ornamentation of the plasma membrane associated with spine-like bodies. The maximum number of cortical microtubules and the external ornamentation were not located at the anterior extremity of the spermatozoon but at a more posterior level, as in all the species of Lepocreadioidea studied to date. Differences observed between spermatozoa of B. plicitum and B. arabicum were observed in: (1) the location of the first mitochondrion, (2) size of cortical microtubules which were longer in B. arabicum than in B. plicitum, and (3) the order of the disappearance of the axonemes, mitochondrion, and cortical microtubules toward the posterior region of the spermatozoon. Only the nucleus reaches into the posterior extremity of the spermatozoon in these two species. The general pattern of the spermatozoon described in these two species exhibit the general model in the genus Bianium.

Anisakid nematodes (Nematoda: Anisakidae) from the marine fishes Plectropomus laevis Lacépède (Serranidae) and Sphyraena qenie Klunzinger (Sphyraenidae) off New Caledonia, including two new species of Hysterothylacium Ward & Magath, 1917.

Based on light and scanning electron microscopical studies, two new species of Hysterothylacium Ward & Magath, 1917 (Nematoda: Anisakidae) are described from the digestive tract of perciform fishes off New Caledonia: H. alatum n. sp. from Plectropomus laevis (Lacépède) (Serranidae) and H. sphyraenae n. sp. from Sphyraena qenie Klunzinger (Sphyraenidae). The former species (H. alatum) is mainly characterised by its large body (male 42.05 mm, gravid females 51.18-87.38 mm long), the shape of the dorsal lip, conspicuously broad cervical alae, a short caecum and a long ventricular appendix, the length of the spicules (925 µm), the number (25 pairs) and distribution of the genital papillae and the tail tip bearing numerous minute cuticular protuberances. The other species (H. sphyraenae) is mainly characterised by the presence of narrow lateral alae, a short caecum and a long ventricular appendix, the length (762-830 µm) and shape of the spicules, the number (37-38 pairs) and arrangement of the genital papillae, and by the tail tip which lacks any distinct cuticular projections visible under the light microscope. In addition, and unidentifiable at the species level, conspicuously large (45.71-66.10 mm long) larvae of Contracaecum Railliet & Henry, 1912, were found in the body cavity of P. laevis, which serves as a paratenic host for this parasite.

Huffmanela hamo sp. n. (Nematoda: Trichosomoididae: Huffmanelinae) from the dagger-tooth pike conger Muraenesox cinereus off Japan.

Huffmanela hamo sp. n. is described from eggs only, which were found in black spots in the somatic musculature of a dagger-tooth pike conger, Muraenesox cinereus (Forsskål), caught off Japan. The eggs are 66-77 microm (mean 72 microm) in length and 33-38 microm (mean 35 microm) in width. The surface of the eggs is smooth and bears neither envelope nor filaments. The species is distinguished from other members of the genus by the dimensions of its eggs and the characteristics of their surface. This is the first species of Huffmanela Moravec, 1987 to be described from an anguilliform fish, and the twentieth nominal species in the genus. Similar black spots with eggs were reported four times in ten years from this fish caught off Japan; although eggs could not be examined, it is likely that the same species was involved in all cases.