Fine Structure of Spermiogenesis in Eastern Pacific Species of Hagfish (Myxinidae)

Abstract Spermiogenesis in Eptatretus stoutii, E. deani and E. sp. display great similarities. Minor differences appear in the acrosomal structure. The spermatozoan has a long middle piece in which the axonema is surrounded by a sheath, consisting of a very few mitochondria, a feature usually associated with internal fertilization.     

Hagfish phylogeny and taxonomy,with description of the new genus Rubicundus (Craniata,Myxinidae)

A recent phylogenetic analysis of the Myxinidae based on the 16S rRNA gene resulted in synonymization of Paramyxine with Eptatretus. This created homonymy of Paramyxine fernholmi with Eptatretus fernholmi and Paramyxine wisneri with Eptatretus wisneri. In order to resolve this nomenclatural dilemma, we made a more extensive phylogenetic assessment of the Myxinidae and examined the nomenclature of the family. We used 75 sequences (37 of which new for this study) of a 561 bp fragment of the 16S rRNA gene, representing 33 species, and 72 sequences (37 of which new for this study) of a 687 bp fragment of the cytochrome c oxidase subunit I (COI) gene, representing 23 species, to reconstruct the phylogeny of Myxinidae. The monophyly of the subfamily Myxininae, traditionally characterized by having a single pair of external gill openings, was rejected (0.50 Bayesian posterior probability) by the 16S analysis, but supported by the COI and combined COI+16S analyses (0.99 and 0.81 Bpp, respectively). The monophyly of the subfamily Eptatretinae, characterized by having several pairs of external gill openings, was not supported by the 16S analysis and rejected by the COI and combined COI+16S analysis due to the placement of Eptatretus lopheliae as the earliest branch of Myxinidae (0.71 and 0.57 Bpp, respectively). Eptatretus lopheliae and Eptatretus rubicundus formed a monophyletic group and were allocated to a new genus, Rubicundus, characterized by the presence of an elongated tubular nostril and reddish coloration. A new monotypic subfamily, Rubicundinae, was proposed for Rubicundus. The synonymy of the genera Paramyxine and Quadratus with Eptatretus was confirmed. E. fernholmi is renamed Eptatretus luzonicus. Eptatretus wisneri was renamed Eptatretus bobwisneri. Petromyzon cirrhatus Forster, 1801, Homea banksii Fleming, 1822, and Bdellostoma forsteri Müller, 1836 are synonyms, but no type specimens are known to exist. Petromyzon cirrhatus was designated as type species of Eptatretus, conserving present usage. Gastrobranchus dombeyi Shaw, 1804 has priority over other names for Chilean myxinids. Bdellostoma stoutii was designated as type species of Polistotrema Gill. The validity of the Western Atlantic Myxine limosa as distinct from the Eastern Atlantic Myxine glutinosa was confirmed.     

Redescription of the hagfishes (Myxinidae: Eptatretus) from southern Africa

The hagfishes of the genus Eptatretus (Myxinidae) from southern Africa are known from three poorly studied species: Eptatretus hexatrema, a common species from Namibia and South Africa; Eptatretus profundus, known only from the holotype collected off Cape Point (South Africa); and Eptatretus octatrema, known from two syntypes from the Agulhas Bank (South Africa). Taxonomic, morphological and distributional information about these three species are reviewed and updated based on the examination of additional specimens collected in South African waters. Eptatretus hexatrema differs from all congeners by having six pairs (rarely seven) of gill apertures arranged in a straight line, 3/2 multicusp pattern of teeth, total cusps 44–49, trunk pores 53–60, total pores 93–107, preventral length 45.1–57.4% TL, tail length 11.6–14.3% TL, tail depth 5.7–8.1% TL, and two bilaterally symmetrical nasal-sinus papillae. Eptatretus octatrema differs from all congeners by having usually eight (some specimens with seven) pairs of gill apertures arranged in a straight line, 3/2 multicusp pattern of teeth, 42–46 total cusps, 22–26 prebranchial pores, 63–68 trunk pores, 104–117 total pores, and two bilaterally symmetrical nasal-sinus papillae. Eptatretus profundus differs from all congeners by having five pairs of gill apertures arranged in a straight line, 3/2 multicusp pattern of teeth, total cusps 42–46, prebranchial pores 12–15, branchial pores 4–5, trunk pores 48–52, tail pores 15–17, total pores 81–86, and body depth at PCD 7.0–9.7% TL. An identification key for the hagfishes from southern Africa is provided and the conservation status of E. octatrema, a species considered to be Critically Endangered, is discussed in light of the new findings.     

Eptatretus eos: A new species of hagfish (Myxinidae) from the Tasman Sea

A new species of hagfish (Myxinidae),Eptatretus eos, is described on the basis of one specimen from the Tasman Sea, west of New Zealand. This five-gilled, deep-sea species differs from previously described hagfish in having the unpaired nostril prolonged into a tube-like snout and in being uniformly pink.     

Spermiogenesis in the Hagfish Eptatretus burgeri (Agnatha)

The fine structure of spermatid differentiation in a primitive vertebrate, the hagfish, whose spermatozoa bear acrosomes, was investigated. In early round spermatids, the acrosomal vesicles were spherical and located in a shallow nuclear indentation, flanked by the plasma and the nuclear membranes. The vesicle underwent a transition through lens-shaped and cap-shaped stages until it attained the shape of a bell in mature spermatozoa. Electron-dense acrosomal material that appeared as deposits in three portions of the vesicle finally joined in the center region at a late stage. Condensation of chromatin occurred in the anterior region of the nucleus. During transformation of the spermatids, many regularly spaced microtubules appeared beneath the plasma membrane except in the anteriormost region of the cell. The microtubules in a single alignment lay parallel to one another and encased the nucleus diagonally. During an early stage, the centrioles changed their orientation from perpendicular to longitudinal and rotated to become parallel to the long axis of the nucleus. Thus, the flagellum lay nearly straight along the cell axis. A cytoplasmic canal appeared transiently during the early stage. A droplet of cytoplasm was eliminated after descending along the flagella. The features of spermiogenesis in hagfish, which lies between invertebrates and vertebrates, are compared with those of other animals.     

Fine Structure of the Surface of Small Mucous Cells in the Epidermis of the Hagfish Myxine glutinosa (Cyclostomata)

Scanning and transmission electron microscopic investigations of the epidermal surface cells, the small mucous cells, of Myxine glutinosa reveal specific structural differentiations in the form of vertical apical channels with microvilli and supporting tonofilament skeleton. The functional significance of these surface differentiations with regard to the living environment of Myxine is discussed and a comparison with the surface characteristics of other aquatic animals is drawn.     

A new species of hagfish (Myxinidae: Eptatretus) from Papua New Guinea

A new species of seven‐gilled hagfish Eptatretus astrolabium (Myxinidae) is described from a 400 mm total length female trapped 1 km east from Planet Rock, Astrolabe Bay, Papua New Guinea, at c. 500 m depth. This is the first hagfish species reported from the waters around New Guinea. It can be distinguished from other hagfishes by a combination of characters including seven pairs of gill apertures, three‐cusp multicusps on the anterior and posterior rows of cusps, 10 posterior unicusps, 52 total cusps, 18–19 prebranchial pores, five branchial pores, 48–49 trunk pores, 83–84 total pores and no nasal‐sinus papillae.     

Estrogens in Plasma of the Hagfish,Myxine glutinosa (Cyclostomata)

Concentrations of estrogens in the plasma of Myxine glutinosa were measured in an attempt to determine whether different stages of reproduction are connected with certain levels of steroid concentration. After extraction with diethylether and column chromatography on Sephadex LH-20, the plasma was radioimmunoassayed for estrogens. Samples were subdivided into 9 groups according to the gonadal conditions of the animals. There appears to be a positive correlation in females between plasma estrogen concentration and egg size (taken as an indication of reproductive stage). After ovulation the level of plasma estrogens fell appreciably. Estrogen levels of most of the samples from male animals were also determined.     

Myxine formosana, a new species of hagfish (Myxiniformes: Myxinidae) from the southwestern waters of Taiwan

Myxine formosana, a new hagfish species, is described on the basis of the specimens from the Pacific Ocean southwest of Taiwan at depths 588–1500 m. It is five-gilled and white-headed with a three-cusp multicusp on the anterior set of cusps and a two-cusp multicusp on the posterior set. Myxine formosana, M. circifrons, M. mccoskeri, and M. robinsi are superficially rather similar, but M. formosana has low to vestigal caudal finfolds. It is the first record of the genus Myxine from Taiwan. Received: October 25, 2000 / Revised: March 17, 2001 / Accepted: April 21, 2001     

Panbiogeographical analysis of distribution patterns in hagfishes (Craniata: Myxinidae)

Aim To analyse the worldwide distribution patterns of hagfishes using panbiogeographical track analysis, and to attempt to correlate these patterns with the tectonic history of the ocean basins. Location Atlantic and Pacific oceans. Method The distributions of 47 out of 70 species of hagfish (in the genera Eptatretus, Myxine, Nemamyxine, Neomyxine, and Paramyxine) were studied by the panbiogeographical method of track analysis. The analysis was performed using distributional data obtained from the collections included in the Ocean Biogeographic Information System (OBIS, http://www.iobis.org ) and FishBase ( http://www.fishbase.org ), with additional records from the literature. Individual tracks were obtained for each species by plotting localities and connecting them by minimum‐spanning trees. Generalized tracks were determined from the spatial overlap between individual tracks. Results Six generalized tracks were found: in the Gulf of Mexico, Caribbean Sea, South‐eastern Atlantic, Western Pacific, North‐eastern Pacific and South‐eastern Pacific. Main conclusions The distribution patterns of myxinids are marked by a high degree of endemism and vicariance, and are correlated with the tectonic features involved in many of the events that led to the development of oceanic basins. The main massing of the group is around the Pacific Basin. In the Atlantic Ocean, the distribution of Myxine glutinosa seems to correspond to a classic trans‐oceanic track and vicariance resulting from the opening of the Atlantic Ocean during the Cretaceous. In the Pacific Ocean, the distribution of the Eptatretus and Paramyxine species is clearly associated with the margins of the Pacific tectonic plate. The generalized tracks of hagfishes are shared by several other groups of marine organisms, including many from shallow tropical waters, implying a common history for this marine biota. Overall, vicariance is a major feature of hagfish distribution, suggesting vicariant differentiation of widespread ancestors as a result of sea‐floor spreading between continents in connection with ocean formation.     

Cyclical Changes in the Testis of the Hagfish Eptatretus burgeri (Cyclostomata)

The development of the spermatogenetic stages in the testis of the hagfish Eptatretus burgeri was studied by means of histological techniques. These animals have an annual reproductive cycle with a spawning season in October. Spermatocytes begin to appear in January and reach a maximum in March. The development of spermatids of stage I and stage II starts in April and June, respectively. In August about 35% of the testicular follicles contain spermatids of stage II. The gonadosomatic index shows a more or less constant increase from November until August. During the spawning season, no animals with well developed testis could be trapped. About 10% of male hagfish bear undeveloped gonads.     

Cyclical Changes in the Ovary of the Hagfish Eptatretus burgeri (Cyclostomata)

Abstract The development of eggs and the degeneration of ovulated follicles in Eptatretus burgeri was studied over a period of one year. It was found that small resting eggs with a diameter of about 1.5 mm continue to grow in November. By the spawning season in October of the following year, the follicles attain a length of about 24 mm. Only one fertilized egg containing an embryo of 7.5 mm length was obtained. The ratio of males to females was found to vary throughout the year. The annual increase of body length in correlation to ovarian development in the hagfish is calculated to be 4 to 5 cm.     

Cardiac Function in the Hagfish,Myxine (Myxinoidea:Cyclostomata)

The electrocardiogram of Myxine glutinosa is described and its events related to systole in the sinus venosus, atrium and ventricle. Intraatrial, ventricular, ventral aortic and dorsal aortic pressures are presented. Ventricular systolic pressure was 10.3 ± 2.0 cm H₂O; average dorsal aortic pressure was 7.0 ± 1.7 cm H₂O. The resistance of the gill vessels caused a loss of 28% of the ventral aortic pressure. The conclusion is drawn that the basic mechanism of the heart of Myxine is like that of fish and higher vertebrates, and that the low pressure it develops is to be ascribed to structural and functional features of the myocardium.     

Eptatretus strickrotti n. sp. (Myxinidae): first hagfish captured from a hydrothermal vent

A single hagfish (Myxinidae, Eptatretus) specimen was recently captured at a hydrothermal vent site on the East Pacific Rise (38 degrees S). This is the first capture of a member of the jawless fishes (agnathans) from a hydrothermal vent site. The specimen differs from all congeners by the very slender body (depth 2.9% of total length), the paired and median ventral nasal sinus papillae, and the presence of 10 afferent branchial arteries on the medial ventral aorta. It is further unique because of a combination of the following features: slime pore counts; paired dorsal nasal sinus papillae; 12 gill pouches and gill apertures; posterior left side of body widely separated from pharyngocutaneous duct; 3/2 multicusp configuration; ventral aorta bifurcated anteriorly between 2nd and 3rd gill pouches (counted from the snout toward the heart); and pink coloration. The specimen is here described as a new species named Eptatretus strickrotti. Molecular 16S rRNA data places this new species as the basal-most species of Eptatretus, providing important new insight to the evolution of hagfishes as a whole.     

Light- and Electronmicroscopic Observations on the Blood Cells of the Atlantic Hagfish,Myxine glutinosa (L.)

In the blood of Myxine glutinosa three cell lines may be distinguished: erythrocytes, granulocytes and agranulocytes. The erythrocytes are remarkably large, oval and nucleated. They are well defined in all their developmental stages by a characteristic micropinocytosis. They originate from blast cells which proliferate in the circulating blood. The blast cells probably form from agranulate stem cells of the intestinal myeloid tissue. The granulocytes constitute about half of the leucocytes. They are neutrophilic with a lobated nucleus. The granulocytopoiesis takes place in the intestinal myeloid tissue. The agranulocytes mainly include two cell types, termed spindle cells and lymphocyte-like cells. These cell types, however, transform into each other. Macrophages occur essentially in the peritoneal cavity rarely in the blood. Transition forms between macrophages and granulocytes may exist. The blood also contains cells which on morphological grounds have been termed thrombocytes. Whether these cells are identical with those necessary for clotting of the blood remains to be proved. With the exception of erythroblasts, the different lines of blast cells are difficult to identify and distinguish from each other. Possibly all lines of blood cells originate from agranulate lymphocyte-like stem cells, most of which are produced by the intestinal myeloid tissue.     

A new species of chondracanthid copepod parasitic in the pharynx of hagfishes (Myxiniformes: Myxinidae) from off Taiwan

A new species of Acanthochondria Oakley, 1927 (Copepoda, Poecilostomatoida, Chondracanthidae), parasitic in the pharynx of hagfishes Eptatretus burgeri (Girard), E. sheni (Kuo, Huang & Mok) and E. yangi (Teng), from off Taiwan is described and illustrated. The new species can be distinguished from its congeners by having the neck region longer than wide (about 2–3 times as long as wide), the moderately long protopod of leg 2 (Type E), the endopod of leg 2 as large as exopod, the cephalosome wider than long, the armature formula of antennule (1, 1, 2, 2, 9), the relatively long trunk (6.7 mm) and egg-sacs (14.4 mm). This is the first record of a parasitic chondracanthid from hagfishes. The findings may provide insights into the phylogenetic relationships and interactions among chondracanthids, hagfishes and demersal fishes, but call for further molecular studies.     

Ultrastructure of the Neurohypophysial Lobe of the Hagfish,Eptatretus stouti (Cyclostomata)

The neurohypophysial lobe is a thin-walled sac that, except for a few blood vessels, lacks any anatomical link with the adenohypophysis. Its wall consists of ependymal, fiber and palisade zones and is surrounded by blood vessels. The lobe is differentiated into distinct dorsal and ventral regions. The dorsal wall is doubly innervated by Gomori-positive axons arising in the anterior hypothalamus and by Gomori-negative fibers of unknown origin. Its surface is covered by an extensive vascular plexus. The ventral wall is innervated only by Gomori-negative fibers and is sparsely supplied with a few fine capillaries. All of the ependymal cells in both regions have the same ultrastructural appearance. The Gomori-positive or Type I axons are identified at the electron microscope level as fibers containing elementary granules with a diameter of 150–230 run. The Gomori-negative or Type II fibers contain dense-cored vesicles that vary from 80–125 nm in diameter. Both Type I and II fibers form synaptic-like complexes with the processes and end-feet of the ependymal cells. Type I axons also abut on the basal lamina bounding the perivascular spaces. It is suggested that the agranular reticulum of the ependymal cells may provide a transport pathway for neural products that are destined for release into the circulation. It is also possible that the ependyma itself is a target of neural activity.     

Review of the Australian hagfishes with description of two new species of Eptatretus (Myxinidae)

This paper revises and updates taxonomic and distributional information about hagfishes (Myxinidae) from Australia. It covers five species of the genus Eptatretus: Eptatretus cirrhatus known from eastern Australia and also distributed around New Zealand, Eptatretus longipinnis endemic to South Australia, Eptatretus strahani originally described from the Philippines and reported here as a new record from Western Australia and two new species described herein as Eptatretus alastairi and Eptatretus gomoni, both from Western Australia. Eptatretus alastairi is distinguished from all congeners by the unique combination of the following characters: six pairs of gill pouches; three‐cusp multicusps on the anterior and posterior rows of cusps; anterior unicusps 9–12; posterior unicusps 8–11; total cusps 48–56; prebranchial pores 13–16; branchial pores 5–6; trunk pores 50–55; tail pores 11–13; total pores 83–88; two bilaterally symmetrical nasal‐sinus papillae in the dorsal surface of the nasal sinus. Eptatretus gomoni is distinguished from all congeners by the unique combination of the following characters: eight pairs of gill pouches; three‐cusp multicusps on the anterior and two‐cusp multicusps on the posterior row of cusps; anterior unicusps 10–11; posterior unicusps 9–10; total cusps 50; prebranchial pores 12–13; branchial pores 7–8; trunk pores 57–58; tail pores 14–15; total pores 91–93; no nasal‐sinus papillae. An identification key for the Australian species of Eptatretus is also provided.     

In Search of the Hagfish Thymus

The Pacific hagfish, Eptatretus stoutii, is capable of a varietyof immunologic responses including allograft rejection and serumantibody production to soluble and cellular antigens. Interesthas revived in the morphology of hagfish lymphoid tissues. Thesearch for a thymus in young specimens has resulted in the discoveryof a phagocytic and antigen-receptive cell population associatedwith the pharyngeal velar muscles. We suggest that a protothymusor precursor of the thymus of higher vertebrates may be containedwithin this velar muscle complex.     

Commensal Nematodes in the Glands,Genitalia, and Brood Cells of Bees (Apoidea)

Seven species of bees from the eastern United States, representing four families in the Apoidea, were dissected and examined for nematode associates. Dufour''s glands in females of Halictus ligatus, Augochlora pura mosieri, and Augochlorella gratiosa (Halictidae) from Florida were infested with dauer juveniles of Aduncospiculum halicti (Diplogasteridae). The Dufour''s glands of Colletes thoracicus (Colletidae) females from Maryland were infested with dauer juveniles of a new species of Koerneria sp. (Diplogasteridae), and abdominal glands of females of Andrena alleghaniensis (Andrenidae) from New York were infested with dauer juveniles of another new species of Koerneria. The lateral and median oviducts, Dufour''s glands, and poison sacs in females of Anthophora abrupta (Anthophoridae) from Maryland and Alabama were infested with dauer juveniles of a new species of Bursaphelenchus sp. (Aphelenchoididae). Cross sections of the nematode-infested poison sacs of A. abrupta revealed two types of humoral host defense reactions.