Larval trematodes infecting the South-American intertidal mud snail Heleobia australis (Rissooidea: Cochliopidae)

Larval trematodes infecting the snail Heleobia australis (Cochliopidae) from the Bahía Blanca estuary, Argentina were surveyed for two years. A total of 7,504 snail specimens was dissected and the larval stages of 15 different trematodes were recovered and examined morphologically. These larvae included four species that had previously been reported from H. australis in Brazil, Uruguay, and Argentina: the heterophyid Ascocotyle (Phagicola) longa and the microphallids, Maritrema bonaerense, Maritrema orensense, and Microphallus simillimus. Three other species, the echinostomatid Stephanoprora uruguayensis, the microphallid Levinseniella cruzi, and the psilostomid Psilochasmus oxyurus are reported here for the first time as parasites of H. australis. Eight other trematodes found in H. australis are described and placed in the appropriate superfamilies, families or genera (Cryptogonimidae, Apocreadiidae, Aporocotylidae, Notocotylidae, Haploporidae, Renicolidae, Himasthla, and Renicola). The prevalence of the trematode taxa infecting H. australis in the Bahía Blanca estuary was low (20%). Microphallidae were the richest and the most prevalent family, probably because of the high abundance of crabs — the second-intermediate hosts of certain microphallid species — and the considerable diversity of gulls. Here we compare the parasite assemblage found in the H. australis from Bahía Blanca estuary with other parasite assemblages infecting Heleobia spp. and other rissooids from the rest of the world.Fil: Alda, Maria del Pilar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Martorelli, Sergio Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; Argentin

Proteomic analysis of skin invasion by blood fluke larvae.

BackgroundDuring invasion of human skin by schistosome blood fluke larvae (cercariae), a multicellular organism breaches the epidermis, basement membrane, and dermal barriers of skin. To better understand the pathobiology of this initial event in schistosome infection, a proteome analysis of human skin was carried out following invasion by cercariae of Schistosoma mansoni.Methodology and resultsHuman skin samples were exposed to cercariae for one-half hour to two hours. Controls were exposed to water used to collect cercariae in an identical manner, and punctured to simulate cercarial tunnels. Fluid from both control and experimental samples was analyzed by LC/MS/MS using a linear ion trap in "triple play" mode. The coexistence of proteins released by cercariae and host skin proteins from epidermis and basement membrane confirmed that cercarial tunnels in skin were sampled. Among the abundant proteins secreted by cercariae was the cercarial protease that has been implicated in degradation of host proteins, secreted proteins proposed to mediate immune invasion by larvae, and proteins implicated in protection of parasites against oxidative stress. Components of the schistosome surface tegument, previously identified with immune serum, were also released. Both lysis and apoptosis of epidermal cells took place during cercarial invasion of the epidermis. Components of lysed epidermal cells, including desmosome proteins which link cells in the stratum granulosum and stratum spinosum, were identified. While macrophage-derived proteins were present, no mast cell or lymphocyte cytokines were identified. There were, however, abundant immunoglobulins, complement factors, and serine protease inhibitors in skin. Control skin samples incubated with water for the same period as experimental samples ensured that invasion-related proteins and host protein fragments were not due to nonspecific degeneration of the skin samples.ConclusionsThis analysis identified secreted proteins from invasive larvae that are released during invasion of human skin. Analysis of specific host proteins in skin invaded by cercariae served to highlight both the histolytic events facilitating cercarial invasion, and the host defenses that attempt to arrest or retard invasion. Proteins abundant in psoriatic skin or UV and heat-stressed skin were not abundant in skin invaded by cercariae, suggesting that results did not reflect general stress in the surgically removed skin specimen. Abundant immunoglobulins, complement factors, and serine protease inhibitors in skin form a biochemical barrier that complements the structural barrier of the epidermis, basement membrane, and dermis. The fragmentation of some of these host proteins suggests that breaching of host defenses by cercariae includes specific degradation of immunoglobulins and complement, and either degradation of, or overwhelming the host protease inhibitor repertoire

Cercariae (Digenea: Strigeidae, Diplostomidae) in Biomphalaria straminea (Planorbidae) from a rice field in Northeastern Argentina

The rice fields can provide habitats for many species of aquatic invertebrates, as insects, molluscs, crustaceans; and vertebrates, as fish, amphibians, reptiles and birds, which may act as hosts in the life cycles of digenean parasites. In this context, the goal of the present study was to describe the cercariae found in Biomphalaria straminea from a rice field in Northeastern Argentina. This freshwater snail inhabits a wide variety of environments, favoring shallow, temporary and standing or slow-flowing freshwater bodies. For this study, snails were collected from the cultivated parcels and irrigated channels during the flooding periods (from the time of sowing to soon after harvesting of the rice) between December 2010 and May 2011 and December 2011 and April 2012, in a rice field from Corrientes province, one of the main rice producers of Argentina. A total of 5 510 snails were examined of which 26 were infected with three different larval trematodes belonging to Strigeidae, Furcocercaria V (0.40 %) and Furcocercaria XX (0.04 %), and Diplostomidae, Furcocercaria XIX (0.04 %). Furcocercaria XX and Furcocercaria XIX were present in one rice cultivation cycle, while Furcocercaria V was present in both rice cultivation cycles. The prevalence of the different furcocercariae was somewhat higher in the second rice cultivation cycle. The cercariae described are new records for Argentina that added to 53 cercariae previously reported for Biomphalaria spp. in different aquatic environments of country. The study of the digenean larval in rice fields is important because contribute to the knowledge of the biodiversity of these environments.Fil: Fernández, María Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Centro de Ecología Aplicada del Litoral. Universidad Nacional del Nordeste. Centro de Ecología Aplicada del Litoral; ArgentinaFil: Hamann, Mónika Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Centro de Ecología Aplicada del Litoral. Universidad Nacional del Nordeste. Centro de Ecología Aplicada del Litoral; Argentin

Uptake of macromolecules by cercariae during skin penetration and transformation to schistosomula (Schistosoma mansoni)

Here, we observed the uptake of membrane-impermeant molecules by cercariae as they penetrate the skin and are transformed into schistosomula. We propose that membrane-impermeant molecules, Lucifer Yellow, Propidium iodide and Hoechst 33258 enter the parasite through both thenephridiopore and the surface membrane and then diffuse throughout the body of the parasite. We present a hypothesis that the internal cells of the body of the schistosomulum represent a new host-parasite interface, at which skin-derived growth factors may stimulate receptors on internal membranes during transformation of the cercariae into the schistosomulum

Reconnaissance of the Distribution and Abundance of Schistosomatium Douthitti, a Possible Human Disease Agent in Surface Waters in Alaska

Studies during the summer and early fall of 1967 show that Schistosomatium douthitti, a blood fluke which may pose a health hazard to man, is well established in the surface waters and surrounding terrestrial environments in the Fairbanks area. It is almost certain that this situation exists throughout Interior Alaska. Ecologically and geologically, the lakes and ponds in which it has been found are the most abundant types in the Interior and both the specific lakes and the types which they represent are abundantly used by man. The life cycle of the worm in this area is probably sustained mostly in small mammals, especially in Microtus pennsvlvanicus but also in Clethrionomys rutilus. The infection certainly over-winters in the mammal host but probably also survives in the snail host under the ice. Although the fluke was only found in two of the nine mammalian species examined, it is probable that it occurs in other than Microtus pennsvlvanicus and Clethrionomys rutilus.The work upon which this report is based was supported in part by funds provided by the U. S. Department of the Interior, Office of Water Resources Research, as authorized under the Water Resources Research Act of 1964. Project Number: A-O20- ALAS Agreement Number: 14-01-0001-89

Can the studies at a spatial scale of 100s meters detect the spatiotemporal fluctuations of a parasite assemblage?

The importance of small-scale heterogeneity in local factors which overrides that of larger-scales factors, suggest that local factors play a major role in determining the richness and prevalence of larval digeneans in intertidal gastropods. The restricted distribution of the snail Heleobia australis along a 500 m transect in Cangrejo creek (Mar Chiquita, Argentina) provides a good opportunity to test the assumption that a study at spatial scale of 100s meters can detect spatiotemporal fluctuations of a larval digenean assemblage. To analyze that, 3600 specimens of H. australis were collected seasonally during the year 2011. A quantitative variation and a space-time interaction between sampling points and seasons of the year for the total prevalence of larval digeneans and snail’s densities were found, as well as a positive correlation with abiotic factors. These results revealed that the fluctuations in the community of larval digeneans of the snail H. australis can be detected at small spatial scale, using its natural distribution of 500 m. This study also highlights the importance of seasonality as a factor that must be considered in studies focused on the search for patterns structuring the communities of larval digeneans, at medium and large scales.Fil: Parietti, Manuela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Merlo, Matias Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Etchegoin, Jorge Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; Argentin

Stunting of the penis in Heleobia parchappii (Mollusca: Cochliopidae) and its relationship with parasitism

Penis anatomy is used to discriminate species of gastropods belonging to the family Cochliopidae; however, this characteristic may be affected by the presence of parasites. To evaluate the possible effect of parasites on penis length and number of papillae in Heleobia parchappii, 195 males were collected from the Nahuel Rucá Lagoon, Argentina. Male snails were only infected by trematode digeneans (total prevalence 45.13%). Three out of 9 species of digeneans registered showed prevalence values higher than 10%: Microphallus szidati, M. simillimus, and Notocotylidae sp. 1. The penis length of non-parasitized males and those parasitized by M. szidati and M. similimus increased with increased snail length; however, this increase was lower in infected snails. In the case of snails infected with Notocotylidae sp. 1, no relationship between shell length and penis length was apparent. Differences in the life cycles of these 3 digeneans could explain the null or lower penis growth rate in relation to host body growth. In contrast, no change was observed in the number of penial papillae of H. parchappii when these snails were infected by larval digeneans compared to those that were not infected. This indicates that penial papillae may be a more stable characteristic than penis length to discriminate between species within the Cochliopidae. The study of penial papillae should be central in the taxonomy and identification of new species within the Cochliopidae, as well as in previously described species.Fil: Merlo, Matias Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología. Laboratorio de Parasitología; ArgentinaFil: Parietti, Manuela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología. Laboratorio de Parasitología; ArgentinaFil: Etchegoin, Jorge Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología. Laboratorio de Parasitología; Argentin

<i>Schistosoma mansoni</i> cercariae experience influx of macromolecules during skin penetration

We have observed that when cercariae penetrate the skin of mice, there is influx into their tissues of Lucifer Yellow and certain labelled molecules of up to 20 kDa molecular weight. This observation was made using a variety of fluorescent membrane-impermeant compounds injected into the skin before the application of cercariae. This unexpected phenomenon was investigated further by transforming cercariae in vitro in the presence of the membrane-impermeant compounds and examining the distribution by microscopy. In schistosomula derived from this procedure, the nephridiopore and surface membrane were labelled while the pre- and post-acetabular glands were not labelled. The region associated with the oesophagus within the pharyngeal muscle clearly contained the fluorescent molecules, as did the region adjacent to the excretory tubules and the germinal mass. We used cercariae stained with carmine to aid identification of regions labelled with Lucifer Yellow. Although the mechanism of this influx is unclear, the observation is significant. From it, we can suggest an hypothesis that, during skin penetration, exposure of internal tissues of the parasite to external macromolecules represents a novel host-parasite interfac

Characterization, cloning and immunogenicity of antigens released by transforming cercariae of Schistosoma mansoni

A schistosome infection is initiated when the parasite penetrates the skin of a susceptible host. Relatively large quantities of protein are released by transforming cercariae compared to later larval stages. This represents the first parasite material to which the host's immune system is exposed, yet little is known about the proteins which are released during the first few hours post-transformation. We have shown that antiserum raised against such molecules was capable of imparting protection against a schistosome challenge infection upon passive transfer to naïve mice. By screening a cercarial cDNA library with this serum, 38 positive clones were identified. Sequence analysis showed these to represent 8 different molecules which included Schistosoma mansoni 21·7 kDa antigen, calcium-binding-protein and the vaccine candidate glutathione S-transferase (Sm28GST). In addition, 5 clones were isolated, 1 of which had significant homology to many cytochrome C proteins, another with leukocyte elastase inhibitors and 3 which represented novel molecules. Four clones were expressed in a prokaryotic high-level expression vector, sera produced against each purified recombinant protein and used subsequently to probe Western blots and parasite sections. The leukocyte elastase inhibitor homologue and 2 unknowns induced significant proliferation by lymph node cells recovered from mice vaccinated with irradiated cercariae. More strikingly, the 2 novel proteins stimulated very high levels of interferon [gamma] (IFN[gamma]) secretion both by lymph node cells and those recovered by broncho-alveolar lavage from the lungs of vaccinated mice. Such results will be discussed in the context of vaccine development