On the structural organization of the bacillary band of Trichuris muris under cryopreparation protocols and three-dimensional electron microscopy.

Whipworms of the genus Trichuris are nematode parasites that infect mammals and can lead to various intestinal diseases of human and veterinary interest. The most intimate interaction between the parasite and the host intestine occurs through the anterior region of the nematode body, inserted into the intestinal mucosa during infection. One of the most prominent structures of the nematode surface found at the infection site is the bacillary band, a surface domain formed by a number of cells, mostly stichocytes and bacillary glands, whose structure and function are still under debate. Here, we used confocal microscopy, field emission scanning electron microscopy, helium ion microscopy, transmission electron microscopy and FIB-SEM tomography to unveil the functional role of the bacillary gland cell. We analyzed the surface organization as well as the intracellular milieu of the bacillary glands of Trichuris muris in high pressure frozen/freeze-substituted samples. Results showed that the secretory content is preserved in all gland openings, presenting a projected pattern. FIB-SEM analysis showed that the lamellar zone within the bacillary gland chamber is formed by a set of lacunar structures that may exhibit secretory or absorptive functions. In addition, incubation of parasites with the fluid phase endocytosis marker sulforhodamine B showed a time-dependent uptake by the parasite mouth, followed by perfusion through different tissues with ultimate secretion through the bacillary gland. Taken together, the results show that the bacillary gland possess structural characteristics of secretory and absorptive cells and unequivocally demonstrate that the bacillary gland cell functions as a secretory structure.

Boehmiella wilsoni (Nematoda, Heligmosomoidea, Boehmiellidae fam. nov.), found in Amazonian rodents.

The genus Boehmiella was initially described as a member of the family Trichostrongylidae. Subsequently, it was assigned to the subfamily Haemonchinae in the family Haemonchidae. We analyzed parasites of spiny tree-rats, Mesomys hispidus, collected in the Amazon rainforest, which were identified as B. wilsoni based on integrative taxonomy. Using morphology, morphometry, and scanning electron microscopy (SEM), we added new data to the original description of the species. We also inferred phylogenetic hypotheses for its relationships within the Trichostrongylina, based on partial nuclear 18S and 28S rRNA genes, through Maximum Likelihood and Bayesian analyses. In conclusion, B. wilsoni does not belong to the family Haemonchidae, nor is it closely related to any other trichostrongylin family, and therefore, we propose the establishment of a new family, Boehmiellidae fam. nov., to which the genus Boehmiella is allocated.

Eurytrema coelomaticum: updated morphology of adult worms using advanced microscopy experiments.

Eurytrema coelomaticum is a digenean flatworm of ruminants that is the causative agent of eurytrematosis, a disease of veterinary health concern. Although modern techniques of morphological analysis have provided new insights about the morphology and anatomy of parasitic helminths, most studies on E. coelomaticum adults are based on conventional light microscopy. In the present study, a combined approach using brightfield, fluorescence, confocal and scanning electron microscopies (SEMs), together with the cryofracture technique, have updated morphological data on E. coelomaticum recovered from cattle in Rio de Janeiro State, Brazil. Light microscopy confirmed the presence of several structures present in the current description, such as suckers, pharynx, oesophagus, intestinal bifurcation and the cirrus-sac. Fluorescence stereomicroscopy revealed for the first time the cubic crystal protein inclusions in the forebody, which were further detailed by confocal and SEMs. Confocal microscopy provided detailed information of the muscular architecture associated with the attachment structures (suckers), digestive system (pharynx and oesophagus), egg-forming complex (ovary, Mehlis' gland and Laurer's canal) and male reproductive system, which are similar to those found in other digenean flukes. SEM images of cryofractured parasites showed mucus and developing eggs within uterine loops. It was demonstrated that the combination of advanced tools generated complementary information, confirming the importance of experimental morphology in parasitology. Therefore, the knowledge of the adult structural organization of E. coelomaticum was improved and this work has contributed to propose new morphological criteria to evaluate the effects of antiparasitic drugs on flukes of medical and veterinary importance.

Xiphidiocercariae from naturally infected Lymnaea columella (Mollusca, Gastropoda) in urban area: morphology and ultrastructure of the larvae and histological changes in the mollusc host / Xifidiocercárias obtidas em Lymnaea columella (Mollusca, Gastropoda) naturalmente infectadas em área urbana: morfologia e ultraestrutura da larva e alterações histológicas do molusco hospedeiro

Abstract Urban agriculture is very common in the cities, as Rio de Janeiro and adjacencies, being important as economic activity. However, this practice can create appropriate conditions for establishment and maintenance of life cycle of many parasites. Lymneid snails can act as intermediate hosts of many trematodes species, including those that infect humans and wild and domestic. In the present study, the snail Lymnaea columella was collected from an urban watercress plantations and the morphology of cercariae released by them was analyzed by light and scanning electron microscopy. The specimens were identified as xiphidiocercariae, belonging to the Cercariae Ornatae group, of the order Plagiorchiiformes. Histological examination of the digestive gland of the infected snails revealed the presence of developing sporocysts, with hemocitary infiltration and metaplasia in the epithelial tissue.

Resumo Agricultura urbana é muito comum nas cidades, como Rio de Janeiro e adjacências, sendo uma importante atividade econômica de subsistência. No entanto, esta prática pode criar condições adequadas para o estabelecimento e manutenção do ciclo de vida de muitos parasitos. Os moluscos limneídeos podem atuar como hospedeiros intermédios de muitas espécies de trematódeos, incluindo aqueles que infectam humanos e animais selvagens e domésticos. No presente estudo, o molusco Lymnaea columella foi coletado de plantações urbanas de agrião e a morfologia das cercárias liberadas por eles foi analisada por microscopia eletrônica de varredura leve e de varredura. As larvas foram identificadas como xifidiocercárias, pertencentes ao grupo das Cercariae Ornatae, Ordem Plagiorchiiformes. Análises histológicas da glândula digestiva dos moluscos infectados evidenciaram esporocistos, com formação de granuloma e metaplasia do tecido epitelial.

Taxonomy of Physaloptera mirandai (Nematoda: Physalopteroidea) based in three-dimensional microscopy and phylogenetic positioning.

Gastrointestinal nematodes are important ecological assets for the maintenance of the biodiversity in the Atlantic Forest in Brazil. They parasitize a number of animals of the local fauna, in which some species can promote serious injuries in the stomach wall of their hosts, which may lead to death. Among these nematodes, parasites of the genus Physaloptera are known to parasitize mammals (particularly carnivores and small rodents), birds and reptiles, being important for the local biodiversity. In this work, three hundred and sixty-two nematodes were recovered from the stomach of twenty-one Metachirus nudicaudatus (Didelphimorphia: Didelphidae) collected in Duas Bocas Biological Reserve, State of Espírito Santo, one of the largest Atlantic Forest remnants and important wildlife refuge of the Atlantic Forest in Brazil. Analysis using fluorescence and scanning electron microscopy as well as phylogenetic assessment using the mitochondrial cytochrome c oxidase subunit I gene showed that the parasites belong to the Physaloptera. Our results show details of the nematode morphology including the cloacal papillae distribution, cuticular topography details, 2D and 3D measurements of the structures with taxonomic importance. Molecular data confirmed the validity of P. mirandai and the phylogeny supported the monophyly of the assemblage formed by Physaloptera and Turgida. The use of a combination of quantitative and multidimensional microscopy tools, such as 3D reconstruction and modeling, allied to phylogenetic analysis may provide grounds for a new approach on helminth taxonomy and structural characterization.

Xiphidiocercariae from naturally infected Lymnaea columella (Mollusca, Gastropoda) in urban area: morphology and ultrastructure of the larvae and histological changes in the mollusc host.

Urban agriculture is very common in the cities, as Rio de Janeiro and adjacencies, being important as economic activity. However, this practice can create appropriate conditions for establishment and maintenance of life cycle of many parasites. Lymneid snails can act as intermediate hosts of many trematodes species, including those that infect humans and wild and domestic. In the present study, the snail Lymnaea columella was collected from an urban watercress plantations and the morphology of cercariae released by them was analyzed by light and scanning electron microscopy. The specimens were identified as xiphidiocercariae, belonging to the Cercariae Ornatae group, of the order Plagiorchiiformes. Histological examination of the digestive gland of the infected snails revealed the presence of developing sporocysts, with hemocitary infiltration and metaplasia in the epithelial tissue.

Spirurids from Gracilinanus agilis (Marsupialia: Didelphidae) in Brazilian Pantanal wetlands with a new species of Physaloptera (Nematoda: Spirurida).

Gastrointestinal nematodes were recovered from thirty four Gracilinanus agilis from forty four collected in Pantanal, Mato Grosso do Sul State, Brazil. Two hundred seventy four spirurids were recovered from the esophagus, stomach and intestines, comprising three species from three different genera. These were identified as Pterygodermatites (Paucipectines) jägerskiöldi, Spirura guianensis and Physaloptera herthameyerae n. sp. is first described. This is the first record of nematodes of the genera Physaloptera and Spirura in hosts of the genus Gracilinanus. The high prevalence of spirurids in 72.3% of the G. agilis collected probably is influenced by the arboreal and diet behaviors.