First report of neurotoxic effect of the cyanobacterium Cylindrospermopsis raciborskii on the motility of trematode metacercariae.

Cylindrospermopsis raciborskii (Woloszynska) is a photosynthetic cyanobacterium that can produce cytotoxic (cylindrospermopsin) and neurotoxic cyanotoxins (saxitoxins). In Brazil the strains of C. raciborskii are reported to produce only saxitoxins (STX) and their effect on fish parasites has not been tested to date. The fish Poecilia vivipara Bloch and Schneider is a common host for the trematode Pygidiopsis macrostomum Travassos off the coast of Rio de Janeiro, and this fish-parasite interaction is a model for behavioural and ecotoxicological studies. The aim of this work was to evaluate the motility of metacercariae of P. macrostomum from P. vivipara exposed to 40 mg l-1 and 400 mg l-1 of crude lyophilized extract of the cyanobacterium C. raciborskii (CYRF-01) for 48 h. The fish were separated into groups of ten individuals and, after exposure, five fish from each group were dissected for counting and checking the motility of metacercariae. The other five fish were dissected after 48 h in clean water. The detection and quantification of STX in the solutions of cyanobacteria, and the gills and guts of fish, were performed by an enzyme-linked immunosorbent assay. The crude extract of C. raciborskii caused temporary paralysis in metacercariae of P. macrostomum after exposure of fish to both concentrations, and the motility recovered after the fish were kept for 48 h in clean water. STX was detected in the guts and gills of all fish analysed, suggesting that this toxin is involved in the paralysis of metacercariae. This is the first report on the action of neurotoxins in metacercariae of fish.

Molecular characterization and confocal laser scanning microscopic study of Pygidiopsis macrostomum (Trematoda: Heterophyidae) parasites of guppies Poecilia vivipara.

Pygidiopsis macrostomum and Ascocotyle (Phagicola) pindoramensis (Digenea: Heterophyidae) parasitize guppies as intermediate hosts and, respectively, fish-eating mammals or birds as definitive hosts. Heterophyids have zoonotic potential, and molecular studies associated with morphological and ecological aspects have helped to clarify their taxonomy and phylogeny. Poecilia vivipara naturally parasitized by metacercariae of both species (100% prevalence) exhibit no external signs of parasitism. In this work, four new sequences of P. macrostomum (18S rDNA, 28S rDNA and ITS2 rDNA) and one new sequence of A. (P.) pindoramensis (mtDNA cox-1) are presented. Phylogeny reconstructions linked P. macrostomum to other heterophyids, but the separation of the Heterophyidae and Opisthorchiidae remains unclear. Additionally, we used indirect immunocytochemistry and the phalloidin-fluorescence techniques allied with confocal laser scanning microscopy to describe muscular and neuronal structures of P. macrostomum. A complex arrangement of muscular fibres is associated with the tegument, suckers, gut and reproductive system. Radial fibres around the ventral sucker are thick, branched and extend to the body wall. High-resolution confocal imaging revealed a typical digenean muscular arrangement and important heterophyid morphological traits. These data will support future control measures to reduce the parasitism in guppies reared in fish farming systems, especially for aquarium and experimental purposes.

Morphology and molecular analysis of Oncicola venezuelensis (Acanthocephala: Oligacanthorhynchidae) from the ocelot Leopardus pardalis in Brazil.

Oncicola venezuelensis Marteau, 1977 was found parasitizing adults of Leopardus pardalis (Linnaeus) found dead in Serra da Capivara National Park, Piauí state, Brazil, a new geographical locality reported for the species. The diversity of Oncicola Travassos, 1916 species is large, but genetic data are scarce. This article presents the results of genetic, morphological and ultrastructural studies carried out for taxonomic purposes. The first ultrastructural view showed a globular, short proboscis with 36 hooks, divided into six longitudinal rows of six hooks each. Hooks differ in size and shape: hooks I, II and III have a 'chisel-shaped' tip. The genetic profile included new sequences of ribosomal DNA ITS1, 5.8S and ITS2, and partial 28S rRNA regions. The results of maximum-likelihood tree analyses for each region showed Oligacanthorhynchidae Southwell et Macfie, 1925 close to Gigantorhynchidae Hamann, 1892 (supported >91%). Both use mammals and birds as definitive hosts. Morphological and ultrastructural studies combined with genetic analysis shed more light on the diversity of Oncicola species.

Parasite-induced and parasite development-dependent alteration of the swimming behavior of fish hosts.

Parasites with complex life cycles have the ability to change the behavior of their intermediate host in a way that increases their transmission rate to the next host. However, the level of behavioral changes can vary considerably, depending on the stage of parasite development and parasite intensity. To investigate the influence of such parameters, we evaluated the locomotory activity of the fish Poecilia vivipara prior to experimental infections, 7 days post-infection (dpi) and 14dpi with cercariae of the digenean Ascocotyle (Phagicola) pindoramensis. The locomotory activity was monitored using an image system, Videomex(®), linked to with a video camera able to record the swimming behavior of the fishes. At the end of the experiments, fishes were dissected and all metacercariae from the gills and mesenteries, the specific sites utilized by A. (P.) pindoramensis, were recovered and counted. There was a significant decrease in the swimming behavior of fishes after 14dpi. Similarly, we found a significant correlation between the swimming behavior of the fishes and parasite intensity in both sites of infection. It is surmised that the decrease in locomotory activity of P. vivipara caused by A. (P.) pindoramensis can disturb its predator-prey relationship in natural environment.