Scanning and transmission electron microscopic studies of the surface of Vallisia indica (Monogenea,Polyopisthocotylea)

Ultrastructural studies were carried out on the surface of Vallisia indica, found on the gills of Scomberoides commersonianus in the Indian ocean. Tegumental serrations present on the hump and on the dorsal and ventro-lateral surfaces of the hindbody lie in the same plane as the secondary gill lamellae and may provide anchorage against displacement forces. Elsewhere the tegument is highly folded forming a reticulum of lamellae and microvilli. The surface syncytium of the tegument in the region of the serrations possesses fibrous electron-dense bars, and a continuous fibrous electron-dense terminal web is present beneath the apical plasma membrane in all regions. The haptor consists of two asymmetrical rows of pincer-type clamps and a terminal lappet with a pair of hooks. Each clamp may be withdrawn into a cushion-like pad. Uniciliated sensory structures are found around the mouth and on the body surface, whereas non-ciliated papillae-like structures are present on the haptor and hindbody. A pair of cone-like sensilla is present on the lappet.     

Morphology,microhabitats and geographical variation of Kuhnia spp. (Monogenea: Polyopisthocotylea)

Kuhnia scombri and K. sprostonae are the only species of the genus recognized as valid. Hosts are scombrid fishes and possibly the genus Scomber only. Examination of many populations from localities in the Atlantic and Pacific showed much geographical variation. Variation is greatest in K. sprostonae, with a distinctly different population on S. scombrus in the North Atlantic. There are intermediate stages in all characters between different populations of both species (except possibly K. sprostonae in the North Atlantic). Some populations within each species have different microhabitats on the gills and pseudobranchs. Differences between parasite populations correspond to some degree to those of host populations, but at least some of the differences are non-adaptive.     

Cholinergic, serotoninergic and peptidergic components of the nervous system of Discocotyle sagittata (Monogenea: Polyopisthocotylea)

Cholinergic, serotoninergic (5-HT) and peptidergic neuronal pathways have been demonstrated in both central and peripheral nervous systems of adult Discocotyle sagittata, using enzyme histochemistry and indirect immunocytochemistry in conjunction with confocal scanning laser microscopy. Antisera to 2 native flatworm neuropeptides, neuropeptide F and the FMRFamide-related peptide (FaRP), GNFFRFamide, were employed to detect peptide immunoreactivity. The CNS is composed of paired cerebral ganglia and connecting dorsal commissure, together with several paired longitudinal nerve cords. The main longitudinal nerve cords (Iateral, ventral and dorsal) are interconnected at intervals by a series of annular cross-connectives, producing a ladder-like arrangement typical of the platyhelminth nervous system. At the level of the haptor, the ventral cords provide nerve roots which innervate each of the 8 clamps. Cholinergic and peptidergic neuronal organisation was similar, but distinct from that of the serotoninergic components. The PNS and reproductive system are predominantly innervated by peptidergic neurones.     

Chaetotaxy of members of the Gyrodactylidae (Monogenea), with comments upon their systematic relationships with the Monopisthocotylea and Polyopisthocotylea

This study investigates intra- and inter-generic variation in the chaetotaxy pattern of members of the Gyrodactylidae and compares these data with those from other monogenean groups elucidated by other workers. The staining of surface sensillary structures has revealed areas of homology between the two genera Gyrodactylus and Ooegyrodactylus, which have markedly contrasting modes of reproduction. Although species specific maps have been elucidated for members of the genus Gyrodactylus, no evolutionary trends in their sensillary patterns could be determined. Members of the family Gyrodactylidae are shown to possess a cluster of 3 × 2 sensilla, similar in their size and arrangement to the post-ocular (spo) cluster of sensilla in polyopisthocotyleans, although on the opposite surface. A comparable cluster of sensilla is lacking in monopisthocotyleans. This information gathered from chaetotaxy supports the proposal that the order Gyrodactylidea is close to the polyopisthocotylean line of monogenean evolution.     

The aflagellate spermatozoon of Diplozoon (Platyhelminthes: Monogenea: Polyopisthocotylea): a demonstrative case of relationship between sperm ultrastructure and biology of reproduction

Diplozoon is known to display an exceptional biology of reproduction: the hermaphroditic adults are permanently fused together and their genital ducts communicate. In contrast to all other polyopisthocotylean monogeneans in which the spermatozoa show an homogeneous biflagellate structure, the spermatozoon of Diplozoon is aflagellate. It is filiform, and composed of a cytoplasmic region and a nuclear region. The cytoplasmic region exhibits mitochondria, a well-developed smooth endoplasmic reticulum, and up to 450 longitudinal singlet microtubules. The microtubules show links between them; seen in cross section, they are arranged as rows or polygons. The spermatozoon nuclear region contains the nucleus surrounded by cortical longitudinal microtubules. The spermiogenesis shows no zone of differentiation, a typical structure found in all other parasitic Platyhelminthes. Diplozoon is the first case of aflagellate spermatozoon found in the parasitic Platyhelminthes. The atypical sperm structure is not linked with phylogeny, but is well correlated with the atypical biology of reproduction.     

The complete mitochondrial genome of Pseudochauhanea macrorchis (Monogenea: Chauhaneidae) revealed a highly repetitive region and a gene rearrangement hot spot in Polyopisthocotylea

The complete mitochondrial genome of Pseudochauhanea macrorchis was determined and compared with other monogenean mitochondrial genomes from GenBank. The circular genome was 15,031 bp in length and encoded 36 genes (12 protein-coding genes, two ribosomal RNAs, and 22 transfer RNAs) typically found in flatworms. Structures of the mitochondrial genome were mostly concordant with that known for Microcotyle sebastis and Polylabris halichoeres, but also contained two noted features-a gene rearrangement hot spot and the highly repetitive region (HRR) in major non-coding region (NCR). The gene rearrangement hot spot located between the cox3 and nad5 genes, including a cluster of tRNA genes, nad6 gene and one major NCR. The HRR seemed to be a unique feature of the polyopisthocotylean mitochondrial genomes. In conclusion, the present study provided new molecular data for future studies of the comparative mitochondrial genomics and also served as a resource of markers for the studies of species populations and monogenean phylogenetics.     

Ultrastructure of spermiogenesis, spermatozoa and the tegument in Atriaster sp. (Platyhelminthes, Monogenea, Polyopisthocotylea, Microcotylidae)

Atriaster (Atriaster) sp., a gill parasite in the fish Diplodus cervinus caught near Dakar, Senegal, was studied by transmission electron microscopy. Spermiogenesis is similar to the general scheme in the Cercomeridae and particularly the polyopisthocotylean monogeneans, but shows two remarkable characteristics: (a) the zone of differentiation for each spermatid is separated from the common spermatid cytoplasmic mass by a region without longitudinal microtubules but with many mitochondria, herein termed the pre-zone of differentiation; (b) transverse sections of nuclei in late spermatids are polygonal with concave sides. These two characteristics are provisionally interpreted as autapomorphies, since observations in other species are lacking. The outer layer of the tegument is similar to that of other polyopisthocotylean monogeneans, and has microvilli and three kinds of cytoplasmic inclusions: granules with fibrous contents, granules with homogeneous contents, and elongate electron-dense bodies.     

Morphology and geographical variation of Pseudokuhnia minor n.g., n.comb. (Monogenea: Polyopisthocotylea)

A new genus, Pseudokuhnia, is established for Kuhnia minor, family Mazocraeidae. It differs from Kuhnia in the presence of two dorso-lateral vaginae, supported by surface ridges; differently shaped large genital hooks, large hamuli and ovary; an opisthaptor which is ventral to and well separated from the body proper; and a rudimentary antero-supplementarium of the clamps. The Atlantic population of P. minor differs from the Pacific population mainly in the larger size of the hamuli and the smaller size of the large and small genital hooks.     

Size differences in hamuli of Kuhnia scombri (Monogenea: Polyopisthocotylea) from different geographical areas not due to differences in host size

Hamuli of Kuhnia scombri from Scomber australasicus and S. japonicus from several localities in the Pacific Ocean show marked differences in length. Hamuli from large fish are slightly longer than those from small fish caught in the same geographical area (northern New Zealand). Differences correlated with host size are much less than those associated with geographical area and may be due to infections acquired in different localities. Taxonomic studies of Monogenea must take geographical variation and, to a lesser degree and in at least some species, effects of host size into consideration.     

Treatment of Microcotyle sebastis (Monogenea: Polyopisthocotylea) infestation with praziquantel in an experimental cage simulating commercial rockfish Sebastes schlegeli culture conditions

The antiparasitic efficacy of praziquantel against the blood-sucking polyopisthocotylean Microcotyle sebastis was tested in an experimental cage simulating commercial rockfish-culture conditions. Juvenile rockfish Sebastes schlegeli were separated into 4 pilot net-pens, and the fish in the groups were either fed a control diet (Group C), fed a praziquantel-adsorbed diet (Group F), bathed in 100 ppm praziquantel for 4 min (Group B), or bathed in 100 ppm praziquantel for 4 min and then fed a praziquantel-adsorbed diet (Group BF). The results of the present study indicate that feeding a praziquantel-adsorbed diet significantly reduces the abundance of M. sebastis infestation, and bathing in 100 ppm praziquantel for 4 min is effective for controlling M. sebastis infestation in practical rockfish culture systems.     

Ultrastructural study of spermiogenesis and the spermatozoon of Microcotyle pancerii (Monogenea: Polyopisthocotylea: Microcotylidae), parasite of meagre Argyrosomus regius (Pisces: Teleostei)

The present work deals with the ultrastructure of spermiogenesis and the spermatozoon of Microcotyle pancerii, a gill parasite of meagre Argyrosomus regius collected in Corsican fish farms. Spermiogenesis was rather similar to that observed in other polyopisthocotylean Monogenea. The intercentriolar body was different from that described in digeneans. The nuclear condensation occurred in 2 successive stages. First, during the nuclear migration in the median cytoplasmic process, the nucleus developed a honeycomb-like appearance. Then, after the flagellar fusion, a discontinuous twisting of the chromatin appeared along the nucleus, with this process ending in total nuclear condensation. The structure of the spermatozoon is characterized by 2 axonemes (9 + 1 pattern), a single and continuous field of cortical microtubules, a mitochondrion, and a nucleus. Our findings were compared with various ultrastructural features in order to highlight variability within the group.     

Evolutionary expansion of the Monogenea

The evolutionary expansion of the monogeneans has taken place in parallel with the diversification of the fish-like vertebrates. In this article the main trends in monogenean evolution are traced from a hypothetical skin-parasitic ancestor on early vertebrates. Special consideration is given to the following topics: early divergence between skin feeders and blood feeders; diversification and specialization of the haptor for attachment to skin; transfer from host to host, viviparity and the success of the gyrodactylids; predation on skin parasites and camouflage; colonization of the buccal and branchial cavities; diversification and specialization of the haptor for attachment to the gills; phoresy in gill parasites; the development of endoparasitism and the origin of the cestodes; the success of dactylogyroidean gill parasites; the uniqueness of the polyopisthocotyleans; ovoviviparity and the colonization of the tetrapods. Host specificity has been the guiding force of coevolution between monogeneans and their vertebrate hosts, but the establishment of monogeneans on unrelated hosts sharing the same environment (host-switching) may have been underestimated. Host-switching has provided significant opportunities for evolutionary change of direction and is probably responsible for the establishment of monogeneans on cephalopod molluscs, on the hippopotamus and possibly on chelonians. There are indications that host-switching may be more common in monogeneans that spread by direct transfer of adults/juveniles from host to host. A limitation on the further expansion of monogeneans is the need for water for the dispersal of the infective larva (oncomiracidium).     

The Parapineal Organ of Teleosts

A parapineal organ was found to be present in 21 teleost fishes belonging to 20 different families, but was absent in poecilids and cyprinodontids. The parapineal organ was situated on the left side of the brain and sent a nerve tract to the left habenular nucleus, except in Gadus, where a “parapineal organ” appeared to send a nerve tract into the pineal stalk. The parapineal organ of adult Gasterosteus consisted of glial elements and parapinealocytes. The latter were small neurons which sent off the unmyelinated axons that formed the parapineal tract. A single photoreceptor cell was found in a stickleback parapineal organ.     

The larva of Oculotrema hippopotami (Monogenea: Polystomatidae)

Oculotrema hippopotami Stunkard parasitizes the eye of the hippopotamus and is the only monogenean known from a mammalian host. Eggs from the uterus of O. hippopotami hatch in water in about 30 days at 24 to 26°C. The ciliated larva resembles the larvae of other polystomatid monogeneans, apart from the absence of hamuli and the unequal lengths of the two intestinal caeca.
Up to 62 eggs are stored within the uterus of the mature worm. They are not expelled when the worm is transferred from the eye of the hippopotamus to a tube of water. Development starts when the eggs are placed in fresh-water. There is no development in saline, and no evidence to suggest that larvae develop in situ around the eye, although very small worms have been collected from this site.     

The mitochondrial genome of Polylabris halichoeres (Monogenea: Microcotylidae)

This study presents the complete mitochondrial (mt) genome of Polylabris halichoeres, which is the largest mt genome sequenced of monogeneans so far and the second complete sequence after Microcotyle sebastis from the Microcotylidae. It is basically similar to that of M. sebastis, with the exception of a high level of gene rearrangement located between trnC and trnL((UUR)), a translocation of trnM and trnH, as well as a highly repetitive region (HRR) in the large non-coding region (NCR). We also find a series of trnI pseudogenes (ΨI) and one unknown short open reading frame (ORF) in the large NCR. Although the ORF cannot be unambiguously regarded as an atp8 gene, we cannot rule out the possibility that it has other functional importance, but it need further study in the future.     

The Infundibular Organ of Branchiostoma lanceolatum

The infundibular organ of adult and larval Branchiostoma was studied by means of the electron microscope. The secretion produced by the infundibular cells is released into the ventricular fluid from secretory vacuoles, fusing with the apical plasmalemma and forming a Reissner's fiber in the ventricle. The basal cell processes reach the external limiting membrane but no basal release of secretory material has been observed. No synapses are in contact with the infundibular cells. The organ seems to function autonomously with neither nervous control from the brain tissue nor chemical regulation from the ventricular fluid.