Cristiane Lafetá Furtado Mendonça

First record of molluscs naturally infected with Angiostrongylus cantonensis (Chen, 1935) (Nematoda: Metastrongylidae) in Brazil

Seeking the identification of Angiostrongylus cantonensis as a potential etiological agent of three clinical cases of eosinophilic meningitis, mollusc specimens were collected in the state of Espírito Santo, Brazil. The snails were identified as Sarasinula marginata (45 specimens), Subulina octona (157), Achatina fulica (45) and Bradybaena similaris (23). Larvae obtained were submitted to polymerase chain reaction and restriction fragment length polymorphism diagnosis. Their genetic profile were corresponded to A. cantonensis. Rattus norvegicus experimentally infected with third-stage larvae, developed menigoencephalitis, and parasites became sexually mature in the lungs. Additionally, larvae obtained from A. fulica snails, from São Vicente, state of São Paulo, also showed genetic profiles of this nematode. This is the first record of Brazilian molluscs infected with this nematode species.

Potentiality of Achatina fulica Bowdich, 1822 (Mollusca: Gastropoda) as intermediate host of the Angiostrongylus costaricensis Morera & Céspedes 1971

Samples of Achatina fulica were experimentally infected with Angiostrongylus costaricensis larvae, etiological agent of abdominal angiostrongyliasis, showing that A. fulica is susceptible to the parasite. Achatina fulica may be a risk to urbanization of abdominal angiostrongyliasis presumably due to its high proliferation, continuous dispersion and remarkable adaptation in several Brazilian towns.

Penetration Sites and Migratory Routes of Angiostrongylus costaricensis in the Experimental Intermediate Host (Sarasinula marginata)

The intermediate hosts of Angiostrongylus costaricensis are terrestrian molluscs, mostly of the family Veronicellidae. The present work aimed at clarifying more accurately the sites of penetration and the migratory routes of A. costaricensis in the tissue slugs and at verifying the pattern of the perilarval reaction at different times of infection. Slugs were individually infected with 5,000 L1, and killed from 30 min to 30 days after infection. From 30 min up to 2 hr after infection, L1 were found within the lumen of different segments of the digestive tube having their number diminished in more advanced times after exposition until complete disappearance. After 30 min of exposition, percutaneous infection occurred, simultaneously to oral infection. Perilarval reaction was observed from 2 hr of infection around larvae in fibromuscular layer, appearing later (after 6 hr) around larvae located in the viscera. A pre-granulomatous reaction was characterized by gradative concentration of amebocytes around larvae, evolving two well-organized granulomas. In this work we confirmed the simultaneous occurrence of oral and percutaneous infections. Perilarval reaction, when very well developed, defined typical granulomatous structure, including epithelioid cell transformation. The infection also caused a systemic mobilization of amebocytes and provoked amebocyte-endothelium interactions.

Alteration in the endogenous intestinal flora of swiss webster mice by experimental Angiostrongylus costaricensis infection

The association between worm infections and bacterial diseases has only recently been emphasized. This study examined the effect of experimental Angiostrongylus costaricensis infection on endogenous intestinal flora of Swiss Webster mice. Eight mice aging six weeks were selected for this experiment. Four were infected with A. costaricensis and the other four were used as controls. Twenty eight days after the worm infection, all mice in both groups were sacrificed and samples of the contents of the ileum and colon were obtained and cultured for aerobic and anaerobic bacteria. In the mice infected with A. costaricensis there was a significant increase in the number of bacteria of the endogenous intestinal flora, accompanied by a decrease in the number of Peptostreptococcus spp. This alteration in the intestinal flora of mice infected by the nematode may help to understand some bacterial infections described in humans.

Angiostrongylus costaricensis and experimental infection of Sarasinula marginata II: elimination routes

Angiostrongylus costaricensis intermediate hosts are terrestrial mollusks mostly belonging to the Veronicellidae family. In the present investigation we focused on the mechanisms of larval expulsion from Sarasinula marginata infected with A. costaricensis. Twenty-five mollusks were individually infected with 5000 L1 and sacrificed at 30 min and 1, 2, 4, 6, and 8 h post-infection and at days 1, 2, 4, 5, 6, 8, 10, 11, 12, 14, 15, 16, 20, 21, 22, 25, 26, 28, and 30 post-infection; the mollusks were then fixed and stained. Diverse organs involved throughout the course of the migratory routes of larvae from oral penetration on were specified and the mechanisms of larval access to the fibromuscular layer through the kidney, rectum, and vascular system were defined. The elimination of L3, derived from oral and/or cutaneous infections, appears to depend on granulomas located close to the excretory ducts of mucous cells.

Macro- and microscopic morphology of the reproductive system of Leptinaria unilamellata (d’Orbigny, 1835) (Mollusca: Pulmonata: Subulinidae)

Many tropical terrestrial gastropods, such as Subulinidae, are recognized and classified by their shells, as is the case for Leptinaria unilamellata (d’Orbigny, 1835), a hermaphroditic pulmonate snail restricted to tropical America. We aimed to characterize the morpho-anatomy and histology of the reproductive system of L. unilamellata. We compared the results obtained from L. unilamellata with the available data on the reproductive system of other subulinid species. The main distinctive characters are those of the penial complex, such as the proportion of the length of the penial complex and the length of the free oviduct, presence of a flagellum, site of insertion of the bursa copulatrix duct relative to the site of insertion of the penial complex and presence and extension of the penial sheath.

Ontogeny and morphological variability of shell in populations of Leptinaria unilamellata (d’Orbigny, 1835) (Mollusca, Pulmonata, Subulinidae).

BackgroundRecent studies concerning species of land snails have revealed that the shell morphometrics can provide evidence of the differentiation among populations. In many cases, the morphologic analysis combined with the investigation of molecular variability, can support changes in taxonomy of studied groups. In this sense, the study of shell morphometry during snail development can contribute to the understanding of the structural mechanisms that creates the diversity observed.DescriptionThe morphological and ontogenetic pattern differences were collected among snails from four different populations, kept under the same laboratorial conditions. It was possible to distinguish characteristic shell morphometrics for snails from each population. The snails from Barra Mansa and Floriano, locations with smaller precipitation indexes presented smaller shell aperture values. The results are discussed in terms of the role of the reproductive strategy of this species as a factor determining shell shape.ConclusionsDifferences in growth allometry indicated that the whole shell forming process is different among the populations, not only the final form of the adult’s shell. Some allometry relationships indicated that, during the snails’ development, the increase in shell width is not proportional to the increase of the width and height of the shell aperture. Thus, there is possibly an antagonism between the adoption of K-strategy and protection against desiccation. Since the spire indices of L. unilamellata morphotypes cannot be explained by physical functional aspects, the most likely explanation is the reproductive strategy of this species.

Angiostrongylus costaricensis life cycle in the intermediate host Sarasinula marginata Semper, 1885 (Mollusca: Soleolifera)

1. Laboratorio de Helmintoses Intestinais do Centro de Pesquisas Rene Rachou da Fundacao Oswaldo Cruz, Belo Horizonte, MG, Bra sil. 2. Departamentode Patologia do Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brasil.Apoio financeiro: PAPES 2/FIOCRUZ.Endereco para correspond encia: Dr. Omar S. Carvalho. Centro de Pesquisas Ren e Rachou/FIOCRUZ. Av. Augusto de Lima 1715, Barro Preto, 30190-002Belo Horizonte, MG.Recebido para publica cao em 02/1/2002.

Life history of Subulina octona (Bruguiere) (Gastropoda: Pulmonata: Subulinidae) based on four-year laboratory observations and a comparative histological analysis of egg-retaining and ovoviviparous subulinids

ABSTRACT In the present study, we aimed to characterize the life history of the land snail Subulina octona integrating information on life-history traits and morphology. We also compared the histology of the free-oviduct and spermoviduct of ovoviviparous and egg-retaining species of subulinids. We considered as ovoviviparous the species in which the complete embryonic development as well as egg hatching occurs inside the parent’s body and, at the end of this process, the parent releases juveniles instead of eggs. We considered as egg-retaining the species in which a great part of the embryonic development takes place inside the parent’s body and the eggs laid contain well-developed embryos. The results showed that the free-oviduct of both ovoviviparous and egg-retaining species shows a histological arrangement that confers greater strength to its walls and is probably related to egg retention. The wall of the spermoviduct is formed by pseudostratified columnar epithelium, with cell apical processes (probably cilia), and by underlying secretory cells. In gravid egg-retaining snails, the eggshells appear in close contact with the secretory cells of the spermoviduct. This fact suggests that these cells play a role in eggshell formation. The present study is the first account for histological features of S. octona, Allopeas gracile and Allopeas micra. The life history of S. octona is a combination of long lifespan, early sexual maturity, indeterminate growth and egg retention. Egg retention limits the number of young that can be produced in one reproductive event. However, the reproductive strategy adopted by S. octona, associating egg retention and K-strategism, compensates this cost of retaining eggs, because the reproductive success may be enhanced as a result of the higher survival of juveniles and the possibility of performing several reproductive events during the year.

Evaluation and updating of the Medical Malacology Collection (Fiocruz-CMM) using molecular taxonomy

BackgroundThe Medical Malacology Collection (Coleção de Malacologia Médica, Fiocruz-CMM) is a depository of medically relevant mollusks, especially from the genus Biomphalaria, which includes the hosts of Schistosoma mansoni. Taxonomic studies of these snails have traditionally focused on the morphology of the reproductive system. However, determination of some species is complicated by the similarity shown by these characters. Molecular techniques have been used to try to overcome this problem.DescriptionThe Fiocruz-CMM utilizes morphological and/or molecular method for species’ identification. However, part of the collection has not been identified by molecular techniques and some points were unidentified. The present study employs polymerase chain reaction-based analysis of restriction fragment length polymorphisms (PCR-RFLP) to evaluate the identification of Biomphalaria in the Fiocruz-CMM, correct existing errors, assess the suitability of taxonomic synonyms, and identify unknown specimens. The results indicated that 56.7% of the mollusk specimens were correctly identified, 4.0% were wrongly identified, and 0.4% was identified under taxonomic synonyms. Additionally, the PCR-RFLP analysis identified for the first time 17.6% of the specimens in the Collection. However, 3.1% of the specimens could not be identified because the mollusk tissues were degraded, and 18.2% of the specimens were inconclusively identified, demonstrating the need for new taxonomic studies in this group.ConclusionThe data was utilized to update data of Environmental Information Reference Center (CRIA). These studies demonstrate the importance of using more than one technique in taxonomic confirmation and the good preservation of specimens’ collection.