Abdoulaye J.S. Bakhoum

Spermatological characters of the digenean Rubenstrema exasperatum (Rudolphi, 1819) (Plagiorchioidea, Omphalometridae)

We present the first ultrastructural description of spermiogenesis and of the spermatozoon of Rubenstrema exasperatum (Omphalometridae), an intestinal parasite of Sorex araneus (Soricidae). Spermiogenesis begins with the formation of the differentiation zone delimited at the base by the ring of the arched membranes and bordered by cortical microtubules. This area contains two centrioles associated with striated rootlets and with an intercentriolar body. It also contains the nucleus and numerous mitochondria. The intercentriolar body is made up of seven electron-dense layers. The two centrioles give rise to two free flagella that grow orthogonally to the median cytoplasmic process. Additionally, flagellar rotation is followed by the proximodistal fusion of the flagella with the median cytoplasmic process, while the nucleus and mitochondria migrate along the spermatid. The constriction of the ring of arched membranes gives rise to the young spermatozoon. The mature spermatozoon of R. exasperatum shows several ultrastructural characters found in digenean spermatozoa such as two axonemes, mitochondrion, two bundles of parallel cortical microtubules, nucleus, and granules of glycogen. External ornamentation of the plasma membrane and spinelike bodies are also present in the spermatozoon of this species. In the present study, we produced additional spermatological data concerning a previously unexplored family, and we also compare our data to the existent ultrastructural descriptions within the Digenea in order to assess the use of several characters proposed as phylogenetic tools.

Spermiogenesis and ultrastructure of the spermatozoon of Wardula capitellata (Digenea, Mesometridae), an intestinal parasite of the sparid teleost Sarpa salpa in Senegal.

The spermiogenesis process in Wardula capitellata begins with the formation of a differentiation zone containing two centrioles associated with striated rootlets and an intercentriolar body. Each centriole develops into a free flagellum orthogonal to a median cytoplasmic process. Later these flagella rotate and become parallel to the median cytoplasmic process, which already exhibits two electron-dense areas and spinelike bodies before its proximodistal fusion with the flagella. The final stage of the spermiogenesis is characterized by the constriction of the ring of arched membranes, giving rise to the young spermatozoon, which detaches from the residual cytoplasm. The mature spermatozoon of W. capitellata presents most of the classical characters reported in digenean spermatozoa such as two axonemes of different lengths of the 9 + “1” trepaxonematan pattern, nucleus, mitochondrion, two bundles of parallel cortical microtubules and granules of glycogen. However, some peculiarities such as two lateral expansions accompanied by external ornamentation of the plasma membrane and spinelike bodies characterize the mature sperm. Moreover, a new spermatological character is described for the first time, the so-called cytoplasmic ornamented buttons.

Collyricloides massanae (Digenea, Collyriclidae): spermatozoon ultrastructure and phylogenetic importance.

The spermatological characteristics of Collyricloides massanae (Digenea: Collyriclidae), a parasite of Apodemus sylvaticus caught in France, were studied by means of transmission electron microscopy. The mature sperm of C. massanae presents two axonemes of different lengths with the 9 + “1” pattern of the Trepaxonemata, two bundles of parallel cortical microtubules, external ornamentation of the plasma membrane, spine-like bodies, one mitochondrion, a nucleus and granules of glycogen. An analysis of spermatological organisation emphasised some differences between the mature spermatozoon of C. massanae and those reported in the Gorgoderoidea species studied to date, specially belonging to the families Dicrocoeliidae, Paragonimidae and Troglotrematidae. The ultrastructural criteria described in C. massanae such as the morphology of both anterior and posterior spermatozoon extremities, the association “external ornamentation + cortical microtubules”, the type 2 of external ornamentation and the spine-like bodies would allow us to bring closer the Collyriclidae to Microphalloidea. However, further ultrastructural and molecular studies are needed particularly in the unexplored taxa in order to fully resolve the phylogenetic position of the Collyriclidae.

Ultrastructural study of vitellogenesis in Maritrema feliui (Digenea, Microphallidae)

During vitellogenesis in the microphallid trematode Maritrema feliui, we distinguished four stages: (I) a stem cell stage of the gonial type; (II) an early differentiation stage with the main cell activity concentrated on the initiation of protein synthetic activity and the beginning of shell globule formation; (III) an advanced differentiation stage concentrated on a rapid intensification of protein synthetic activity, the progressive fusion of individual shell globules into large shell globule clusters and the formation of saturated lipid droplets and a small amount of β-glycogen particles in the peripheral cytoplasm, considered as a store of nutritive reserves for the developing embryos; and (IV) the mature vitellocyte. Early vitellocyte maturation is characterised by: (1) an increase in cell volume; (2) extensive development of large, labyrinth-like cisternae of GER that produce proteinaceous granules; (3) the development of Golgi complexes engaged in packaging this material; and (4) a continuous enlargement of proteinaceous granules within vacuoles and their transformation into shell globule clusters composed of the heterogeneous material observed during vitellocyte cytodifferentiation. Mature vitelline cells are very rich in two types of cell inclusions accumulated in large amounts in their cytoplasm: (1) shell globule clusters, which play an important role in eggshell formation; and (2) a few osmiophobic lipid droplets of a saturated nature that undoubtedly represent nutritive reserves for the developing embryos. In addition, there are small numbers of β-glycogen particles in the peripheral cytoplasm of mature vitellocytes of this species. The general pattern and ultrastructure of vitellogenesis in M. feliui greatly resembles those observed in another microphallid trematode, Maritrema linguilla, in other digeneans and in some lower cestodes. Quantitative and qualitative variations in lipids (saturated and unsaturated) and glycogen (α-glycogen rosettes and β-glycogen particles) during platyhelminth vitellogenesis between the different species of trematodes and some lower cestodes are identified and discussed.

Ultrastructural study of sperm cells in Acanthocolpidae: the case of Stephanostomum murielae and Stephanostomoides tenuis (Digenea)

The mature spermatozoa of Stephanostomum murielae and Stephanostomoides tenuis are described by transmission electron microscopy. They present several ultrastructural features previously reported in other digeneans. Their spermatozoa possess two axonemes of different length showing the 9 + ‘1’ trepaxonematan pattern, four attachment zones, two mitochondria (with an anterior moniliform one in S. murielae), a nucleus, two bundles of parallel cortical microtubules, external ornamentation of the plasma membrane, spine-like bodies and granules of glycogen. The main differences between the mature spermatozoon of S. murielae and S. tenuis are the maximum number of cortical microtubules, the morphology of the anterior spermatozoon extremity and the anterior mitochondrion. This study is the first concerning members of the family Acanthocolpidae. The main ultrastructural characteristics discussed are the morphology of the anterior and posterior spermatozoon extremities, antero-lateral electron dense material, external ornamentations, spine-like bodies and number and morphology of mitochondria. In addition, the phylogenetic significance of all these ultrastructural features is discussed and compared to molecular results in order to highlight the complex relationships in the Digenea.

Ultrastructural studies on egg envelopes surrounding the miracidia of Mediogonimus jourdanei Mas-Coma et Rocamora, 1978 (Digenea, Microphalloidea, Prosthogonimidae)

The intrauterine, mature and fully embryonated eggs of the prosthogonimid trematode Mediogonimus jourdanei Mas-Coma et Rocamora, 1978 were examined by means of transmission electron microscopy (TEM), using high pressure freezing, freeze substitution and infiltration with resin techniques. Each embryonated egg is composed of a miracidium surrounded by three envelopes: (1) the egg shell, (2) the outer and (3) inner envelopes. Egg envelopes play an important role in the protection, metabolism, storage of nutritive reserves and the general biology of the M. jourdanei egg. The inner envelope is characterised by large, flattened nuclei, and its syncytial cytoplasm contains a heavy accumulation of glycogen, lipid droplets, mitochondria and large vesicles. These traits indicate that this layer has the features of a metabolically-active syncytial layer with an energy storage capability. In the infective eggs observed before the hatching of the miracidium, areas of so-called “focal cytoplasmic degradation” were frequently observed, which may be involved in the autolytic process of all components of this envelopes.

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.

Parasite fauna of rodents (Murinae) from El Hierro (Canary Islands, Spain): a multidisciplinary approach.

The parasite fauna (protozoa, helminths and insects) of the two most widespread Murinae rodents in El Hierro (Canary Islands, Spain), the black rat (Rattus rattus) and the house mouse (Mus musculus domesticus) was studied. Faunistic, ecological, ecotoxicological data, as well as information on the biology of some nematode parasites of R. rattus are provided. The present work is unprecedented in the Canary Islands, and provides the first data on the parasite biodiversity in Murinae from the archipelago. Concerning to parasitofaunas stands out: a) impoverishment of biodiversity of helminths respect of which have the same hosts in other islands; b) increasing the number of species of Siphonaptera, even compared with flea species that parasitize the same hosts from continental biotopes.

First spermatological study in the Atractotrematidae (Digenea, Haploporoidea): the case of Atractotrema sigani, intestinal parasite of Siganus lineatus

The ultrastructural organization of the mature spermatozoon of the digenean Atractotrema sigani (from Siganus lineatus off New Caledonia) was investigated by transmission electron microscopy. The male gamete of A. sigani exhibits the general morphology described in digeneans with the presence of two axonemes of different lengths showing the 9 + “1” pattern of the Trepaxonemata, a nucleus, two mitochondria, two bundles of parallel cortical microtubules, external ornamentation, spine-like bodies and granules of glycogen. However, the mature spermatozoon of A. sigani has some specific characters such as the morphology of its anterior region and the submembranous electron-dense material. Although similar structures have been reported in some digenean species, the presence of a submembranous electron-dense material describing a complete ring is reported here for the first time in the mature spermatozoon of A. sigani. In addition, sperm characteristics are compared between the Haploporoidea and their supposed close superfamilies, and possible phylogenetic implications of these findings for the Digenea are discussed.

Spermatozoon ultrastructure in two monorchiid digeneans

Spermatological characteristics of species from two monorchiid genera, Opisthomonorchis and Paramonorcheides, have been investigated, for the first time, by means of transmission electron microscopy. The ultrastructural study reveals that the mature spermatozoon of Opisthomonorchis dinema and Paramonorcheides selaris share several characters such as the presence of two axonemes of different lengths showing the 9+“1” pattern of the Trepaxonemata, a nucleus, two mitochondria, two bundles of parallel cortical microtubules, external ornamentation of the plasma membrane, spine-like bodies, granules of glycogen and similar morphologies of the anterior and posterior extremities. The slight differences between the male gamete of O. dinema and P. selaris are the length of the first axoneme and the position of the second mitochondrion. This study also elucidates the general morphology of the spermatozoon in all monorchiid species described so far, which corresponds to a unique spermatozoon type. Other interesting finds concern the spermatological similarities between monorchiid spermatozoa and the mature spermatozoon reported in the apocreadiid Neoapocreadium chabaudi. These similarities allow us to suggest a close phylogenetical relationship between the Monorchiidae and the Apocreadiidae, although more studies are needed, especially in the unexplored taxa.