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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.
Classification and identification of muscle-parasitizing didymozoids found in marine fish is difficult because of their novel parasitism and morphology. Recent sequence analysis has helped, but only seven sequences are available. Therefore, the usefulness of molecular methods for differentiation of muscle-parasitizing didymozoids, as well as genetic differences between the muscle and the other site-parasitizing didymozoids are quite unclear. In the present study, six unidentified didymozoid isolates from the trunk muscles of four marine fish species (Diagramma pictum, Plectorhinchus cinctus, Pagrus major and Cypselurus heterurus) were examined genetically using sequence analysis (18S rDNA, 28S rDNA, ITS-2 and coxI). All isolates were placed phylogenetically as a lineage independent of other site-parasitizing didymozoids at 18S rDNA, ITS-2 and coxI. They were grouped into three distinct lineages. The present and the previous unidentified or identified didymozoids from trunk muscles were found to differ clearly for every host species by sequence analysis, suggesting that muscle-parasitizing didymozoids are host-specific. This report is the first describing the molecular characteristics of muscle-parasitizing didymozoids by sequence analysis targeting the nuclear and mitochondrial DNA loci, which is proposed as a superior method for didymozoid differentiation.
The ultrastructure of the uterus proper of the aspidogastrean Aspidogaster limacoides Diesing, 1835 and two digenean species, Phyllodistomum angulatum Linstow, 1907 (Plagiorchiida, Gorgoderidae) and Azygia lucii (Müller, 1776) (Strigeida, Azygiidae), was examined by transmission electron microscopy (TEM). The uterine epithelial lining of these species is thin, except for the perinuclear region of the epithelial cells. Septate junctions occur between adjacent epithelial cells within the uterine wall. The luminal surface of the cells is elevated into microlamellae, which project into the uterine lumen and cover the entire epithelial lining. Basally the uterine epithelium is attached to a basal matrix, and its supporting layers of muscle fibres are weak and composed of scattered circular muscles. Despite the marked similarity in the ultrastructural pattern of the aspidogastrean and two digeneans studied, there is some degree of variation in the secretory activity of their uterine epithelium. The high level of vesicular exocytotic activity in the epithelial cytoplasm of A. lucii may be associated with differences in egg emission and the subsequent life cycle involving a non-ciliated, non-swimming and non-free-living miracidium, as opposed to the free-swimming miracidium of P. angulatum. The similar nature of the uterine epithelium in all three species studied represents an ultrastructural marker possibly supporting a close phylogenetic relationship between the Aspidogastrea and the Digenea.
Seriolella porosa (Silver warehou) is an important fish resource in the Argentine Sea. No previous studies have been carried out to date on its parasite fauna. The present study aims to (i) identify the digenean trematodes that parasitize Silver warehou, (ii) estimate parasite frequency and abundance, and (iii) establish their relationship with fish size and sex. A total of 100 specimens of S. porosa from San Matías Gulf, Argentina were examined between September and October 2006. Host size and sex were determined. Parasites extracted from the stomach lumen were fixed, conserved in 5% formalin, and stained with Langeron’s hydrochloric carmine. The following parameters were also determined: dispersion coefficient, prevalence, intensity, and parasite abundance. Differences in parasitism were assessed using non-parametric tests. Four adult digenean species were found in the stomach, namely Lecithocladium cristatum, Aponurus laguncula, Elytrophalloides oatesi and Gonocerca cf. phycidis. L. cristatum was not only dominant but also the most prevalent and abundant species. None of the digeneans showed interspecific association with each other and therefore they might follow different patterns of colonization (i.e., different intermediate hosts, seasonal and/or spatial distribution of the infective stages). Significant differences were observed among the digeneans analyzed in respect to the sex of the host. Results from this study show that Silver warehou plays a key role as a final host of a significant number of digeneans in the ecosystem. This study is the first step in an attempt to further determine the larval stages in intermediate hosts in San Matías Gulf.
Maritrema portucalensis sp. nov. (Digenea, Microphallidae) from Aveiro estuary, northern Portugal, is described on the basis of excysted metacercariae obtained from the gill lamellae of the shore crab Carcinus maenas (L.). M. portucalensis sp. nov. most closely resembles Maritrema subdolum Jägerskiöld, 1909, a species that also frequently uses C. maenas as second intermediate host. The new species differs from M. subdolum in having a narrower body at the level of testes and a smaller ovary and testes. Also, the digestive caeca presented various folds along their length, thickening in the terminal region. Moreover, the body surface was fully covered by tegumental spines provided with several teeth. To complement our morphological characterization and to identify M. portucalensis metacercariae, the ITS1 rDNA region of the cysts isolated from C. maenas was sequenced and compared with the corresponding available sequences of digenean trematodes belonging to the family Microphallidae. Alignments revealed 28 base-pair differences between the query-ITS1 and that of M. subdolum, and more importantly a 100% similarity with the sequence of Microphallidae sp. no. 15 cercaria from the snail Hydrobia ulvae. The last observation allowed us to establish an unequivocal association between cercaria known as Microphallidae sp. no. 15, and the metacercaria of M. portucalensis sp. nov. characterized in this work, contributing decisively to the clarification of its life cycle.
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