Parasitism of terrestrial gastropods by medically-important nematodes in Brazil.

An ample variety of parasitic associations are found between mollusks and nematodes, in which the mollusks may act as intermediate, paratenic or definitive hosts. Some free-living nematodes, in particular those of the order Rhabditida, are also found frequently in terrestrial mollusks. The present study reviews the results of the parasitological testing on samples of terrestrial mollusks conducted at the Brazilian National Reference Laboratory for Schistosomiasis and Malacology between 2008 and 2021. The samples were supplied primarily by the public health authorities from the different regions of Brazil, but also by research institutions and general population. The mollusks were processed individually and the obtained larvae were identified from their morphology and, whenever necessary, by molecular analysis. A total of 1,919 service orders were registered during the period, including 19,758 mollusk specimens collected from 23 of the 26 Brazilian states, as well as the Federal District, totalizing 145 municipalities. There was a marked predominance of the synanthropic species that are widely distributed in Brazil-Achatina fulica (87.08%), Bulimulus tenuissimus (4.18%), Bradybaena similaris (2.06%), and Sarasinula linguaeformis (1.50%). Of the 16,750 terrestrial mollusks examined, nematodes were recorded in 1,308 service orders, with the predominance of the superfamily Metastrongyloidea, in 616 service orders. They included Angiostrongylus cantonensis, rat lungworm, which was found in 252 samples, and Aelurostrongylus abstrusus in 145 samples. Free-living nematodes were found in 952 samples, Ancylostoma caninum and Cruzia tentaculata (previously identified as Strongyluris sp.) in one and 275 samples, respectively, and other parasites in 210 samples (not identified). The results highlight the diversity of the associations between nematodes and terrestrial mollusks in Brazil, in particular invasive and synanthropic species, with emphasis on the giant African land snail, Achatina fulica. They demonstrate the prominent role of this species of mollusk in the transmission of medically-important nematodes, which affect the health of both humans and animals, in particular eosinophilic meningitis, which is caused by Angiostrongylus cantonensis. This reinforces the need for more studies, and justify the growing demand for information as well as parasitological diagnosis of this mollusk, given its wide distribution in Brazil and its impact as an urban pest.

Diversity of non-marine gastropods of the Fiocruz Atlantic Forest Biological Station and adjacents urban areas, Rio de Janeiro, RJ, Brasil.

Our main objective was to perform a preliminary survey of the non-marine gastropods at the Estação Biológica Fiocruz Mata Atlântica (EFMA) and its adjacent urban areas, also considering samples from other localities of the Parque Estadual da Pedra Branca received as donation, in Rio de Janeiro, RJ, Brazil. Infection by larval helminths of medical and veterinary importance was also investigated in 348 specimens of five freshwater species and two specimens of the terrestrial species Achatina fulica. In all, 584 samples of molluscs from 34 collection sites were obtained. They represented 31 species classified in 16 families. Fifteen species were exclusive to the EFMA area. In the urban area mainly exotic and/or synanthropic species were found. Some of them were found in the forest border as well. The freshwater Biomphalaria tenagophila, an intermediate host of the trematode that causes schistosomiasis mansoni, was found parasitized by Xiphidiocercarie cercariae and the Afro-asiatic Melanoides tuberculata by Pleurolophocercus cercaria. The finding of endemic and exotic species, including transmitters of parasites, demonstrate the importance of fast surveys such as the present study. However, considering the great diversity of endemic molluscs found in a relatively small part of the EFMA, we highlight the necessity of further additional studies.

First report of the nematode Cruzia tentaculata using molluscs as natural intermediate hosts, based on morphology and genetic markers.

The life cycles of many parasitic nematodes include terrestrial gastropods as intermediate hosts. Over the past few decades, a number of cases of parasitism between molluscs and medically-important nematodes have been reported in Brazil, in particular, those involving the invasive giant African gastropod, Achatina fulica, and zoonoses caused by the nematodes Angiostrongylus cantonensis and Angiostrongylus costaricensis, the etiological agents of neuroangiostrongyliasis and abdominal angiostrongyliasis, respectively. In the present study, larvae found infecting A. fulica, Latipes erinaceus, and Thaumastus taunaisii, from two localities in the Brazilian state of Rio de Janeiro were characterized using light and scanning electron microscopy, and sequences of the 18S rRNA and MT-CO1 genes. Genetic markers allowed to identify the larvae collected in the present study as Cruzia tentaculata, whose adults parasitize didelphid marsupials in the Americas. These findings indicate that both native and non-native gastropods may act as intermediate hosts and represent a previously unnoticed heteroxenous life cycle of C. tentaculata.

Field Application of NIR Spectroscopy for the Discrimination of the Biomphalaria Species That Are Intermediate Hosts of Schistosoma mansoni in Brazil.

Near Infrared Spectroscopy (NIRS) is a spectroscopic technique that evaluates the vibrational energy levels of the chemical bonds of molecules within a wavelength range of 750-2,500 nm. This simple method acquires spectra that provide qualitative and quantitative data on the chemical components of the biomass of living organisms through the interaction between the electromagnetic waves and the sample. NIRS is an innovative, rapid, and non-destructive technique that can contribute to the differentiation of species based on their chemical phenotypes. Chemical profiles were obtained by NIRS from three snail species (Biomphalaria glabrata, Biomphalaria straminea, and Biomphalaria tenagophila) that are intermediate hosts of Schistosoma mansoni in Brazil. The correct identification of these species is important from an epidemiological viewpoint, given that each species has distinct biological and physiological characteristics. The present study aimed to develop a chemometric model for the interspecific and intra-specific classification of the three species, focusing on laboratory and field populations. The data were obtained from 271 live animals, including 150 snails recently collected from the field, with the remainder being raised in the laboratory. Populations were sampled at three localities in the Brazilian state of Rio de Janeiro, in the municipalities of Sumidouro (B. glabrata) and Paracambi (B. straminea), and the borough of Jacarepaguá in the Rio de Janeiro city (B. tenagophila). The chemometric analysis was run in the Unscrambler® software. The intra-specific classification of the field and laboratory populations obtained accuracy rates of 72.5% (B. tenagophila), 77.5% (B. straminea), and 85.0% (B. glabrata). The interspecific differentiation had a hit rate of 75% for the field populations and 80% for the laboratory populations. The results indicate chemical and metabolic differences between populations of the same species from the field and the laboratory. The chemical phenotype, which is closely related to the metabolic profile of the snails, varied between environments. Overall, the NIRS technique proved to be a potentially valuable tool for medical malacology, enabling the systematic discrimination of the Biomphalaria snails that are the intermediate hosts of S. mansoni in Brazil.

An integrative study of the invasive jumping-snail Ovachlamys fulgens (Gastropoda, Helicarionidae) in Rio de Janeiro and its fast spreading in Southeastern and Southern Brazil.

The Japanese invasive jumping snail Ovachlamys fulgens is a pest of ornamental plants and an intermediate host of a nematode that causes eosinophilic meningitis. We expand its distribution to eight municipalities from Rio de Janeiro State, and one locality from the Paraná State, and generated for the first time partial sequences of the cytochrome c oxidase subunit I (COI) gene for Brazilian populations. External morphology, reproductive system, shell, radula, and jaw were also analyzed and described. Twenty-one lots were collected from Rio de Janeiro, Niterói, Magé, Miguel Pereira, Petrópolis, Teresópolis, Nova Friburgo, Bom Jardim and Paraty, in Rio de Janeiro State, and from Foz do Iguaçu, Paraná State. External morphology, shell and reproductive system were typical of O. fulgens, with some peculiarities found in the shell and radula. A single haplotype was found, which was 100% similar to sequences of COI available in GenBank for specimens from Japan and Argentina. The species seems to be adapted to many habitats and be rapidly expanding its distribution in Southeastern and Southern Brazil, and other South America countries. We highlight the importance of monitoring O. fulgens, considering its potential to compete with native mollusks, attack several plants, and be a transmitter of diseases.

Physiology and immunity of the invasive giant African snail, Achatina (Lissachatina) fulica, intermediate host of Angiostrongylus cantonensis.

As one of the most successful invasive land snail species, Achatina (Lissachatina) fulica Bowdich, 1822 has achieved wide global distribution, particularly in (sub)tropical regions, with further dispersal likely due to climate change. This species of giant African snails (up to 17 cm shell length) is a pest that has extensive negative impact on agriculture and can serve as vector for several parasites, including Angiostrongylus cantonensis, a nematode parasite that causes (human) eosinophilic meningitis, an emergent disease. Investigation showed that A. cantonensis infection negatively impacts the metabolism of A. fulica by depleting polysaccharide stores of the intermediate host, compromising the energy balance of the snail. A review of the literature indicates that A. fulica possesses potent innate type immune defenses to counter infection, including phagocytic hemocytes capable of deploying reactive oxygen species and lectins for non-self recognition, a serine protease-dependent coagulation response (not observed in other taxa of gastropods), as well as antimicrobial proteins including achacin, an antimicrobial protein. A recent chromosome level genome assembly will facilitate progressively detailed characterization of these immune features of A. fulica. We strongly encourage further immunological studies of A. fulica, ranging from organismal level to molecular biology to gain better understanding of the A. fulica internal defense response to nematode pathogens like A. cantonensis and the contribution of immune function to the invasiveness of (snail) species. Characterization of immunity of A. fulica, representing the understudied Stylommatophora (panpulmonate landsnails) will also broaden the comparative immunology of Gastropoda.

Compatibility Polymorphism Based on Long-Term Host-Parasite Relationships: Cross Talking Between Biomphalaria glabrata and the Trematode Schistosoma mansoni From Endemic Areas in Brazil.

Sympatric snail populations have been kept in the laboratory since the isolation of the parasite from the field. To evaluate the influence of the intermediate host in the infectivity of S. mansoni, this allopatric strain was compared to two sympatric strains, from different geographical origins, and with different time of maintenance in the laboratory. Snail-trematode compatibility was accessed for a total of nine possible combinations (three snail populations, three schistosome strains), using different charges of parasite: 1, 5, 10, and 15 miracidia/snail. Each S. mansoni strain was characterized according to its infectivity phenotype that reflects the efficiency of their infection mechanism and all B. glabrata populations were characterized according to its (in)compatible phenotype that reflects the level of (un)susceptibility they display. For all host-parasite combinations tested the dose-response relation indicated a trend for an increase in the infectivity of S. mansoni when higher miracidial doses were used. SmRES-2 presented the highest overall infectivity rate, especially in the SmRES-2/BgRES interaction with 15 miracidia/snail. However, SmRES was more infective to BgBAR than SmRES-2, indicating that SmRES strain was more infective at the first contact with this new host than after 2 years of interaction (SmRES-2). BgBAR presented the highest susceptibility to infection. SmRES and SmRES-2 are the same parasite strains. It seems that during these 2 years of interaction, BgBAR acted like a filter and shifted the compatibility polymorphism of the strain SmRES. SmRES-2 became more infective to BgRES (sympatric) than to BgBAR (allopatric), and conversely, SmRES was more infective to BgBAR (allopatric) than to BgRES (sympatric). This interplay suggests that epigenetic mechanisms are prompting these changes. This study concerns with infection of B. glabrata snails from different Brazilian localities with S. mansoni in allopatric and sympatric associations that will partially help in understanding the natural epidemiology of schistosomiasis within natural snail populations in watercourses. This work demonstrates that there is a shift on the compatibility polymorphism profile resulting from sympatric and allopatric interactions of B. glabrata and S. mansoni that constantly change during the time of interaction.

Effects of experimental Angiostrongylus cantonensis infection on the reproductive biology of Biomphalaria straminea and Biomphalaria tenagophila.

Eosinophilic meningoencephalitis is an endemic zoonosis in Southeast Asia and the Pacific Islands, but in recent years, new cases have been reported in various countries outside these regions, including Brazil, where it is considered an emerging disease. In this study, the effect of infection by the nematode Angiostrongylus cantonensis, one of the main etiologic agent of this disease, on the reproductive biology of the planorbid snails Biomphalaria straminea and B. tenagophila was investigated during the pre-patent period. Alterations in the reproductive biology of B. straminea and B. tenagophila were analyzed in laboratory-reared specimens infected by A. cantonensis during 21days; the number of eggs, number of egg masses, number of eggs/mass, number of eggs/snail, viable eggs/snail, survival and galactogen content in the albumen gland were measured. The results indicated the occurrence of initial compensation in reproductive effort in both snail species, but at different moments in the pre-patent period. More specifically, a reduction of 46.53% in the eggs/egg mass ratio in infected B. straminea was observed, a reflection of a 50% decline in the concentration of galactogen contained in the albumen gland. Changes in this parameter were also noted in B. tenagophila, but only at the end of the study period, with a reduction of 15.49%. Histological analyses indicate that changes observed can be explained by the tissue damages caused by the migration and development of the larvae. These results shed more light on the host-parasite relationship and indicate the importance of studying reproductive aspects for efforts to control infected snails. Considering that terrestrial snails can also transmit eosinophilic meningitis (in addition to aquatic mollusks), the data obtained expand knowledge of this host-parasite relationship and provide support for programs to control this zoonosis.

The influence of Angiostrongylus cantonensis (Nematoda, Metastrongylidae) infection on the aerobic metabolism of Biomphalaria straminea and Biomphalaria tenagophila (Mollusca, Gastropoda).

Angiostrongylus cantonensis is considered the main agent responsible for human eosinophilic meningoencephalitis. This parasite has low specificity for mollusk hosts and it can also use aquatic snails as auxiliary hosts. Studies based on the metabolic profile of Biomphalaria spp. infected by A. cantonensis have been conducted to observe parasite-host interactions. In the present study, the glucose content in the hemolymph and glycogen content in the digestive gland and cephalopedal mass of Biomphalaria tenagophila and Biomphalaria straminea experimentally infected by A. cantonensis were evaluated, along with the activity of LDH. The snails were dissected from 6 to 21days after infection to collect the hemolymph and separate the tissues. Decreases of 96% and 6.4% in the glucose content triggered a transition from aerobic to anaerobic metabolism in the two infected snail species, B. straminea and B. tenagophila, respectively. That finding was confirmed by high-performance liquid chromatography. These results indicate that when infected, these snails are able to change their metabolic profile, suggesting a strategy to maintain their homeostatic balance.

Angiostrongylus cantonensis infection in molluscs in the municipality of São Gonçalo, a metropolitan area of Rio de Janeiro, Brazil: role of the invasive species Achatina fulica in parasite transmission dynamics.

The aim of this study was to analyse the infection dynamics of Angiostrongylus cantonensisin its possible intermediate hosts over two years in an urban area in the state of Rio de Janeiro where the presence ofA. cantonensis had been previously recorded in molluscs. Four of the seven mollusc species found in the study were exotic.Bradybaena similaris was the most abundant, followed by Achatina fulica, Streptaxis sp., Subulina octona, Bulimulus tenuissimus, Sarasinula linguaeformis and Leptinaria unilamellata. Only A. fulica and B. similaris were parasitised by A. cantonensis and both presented co-infection with other helminths. The prevalence of A. cantonensis in A. fulica was more than 50% throughout the study. There was an inverse correlation between the population size ofA. fulica and the prevalence of A. cantonensis and abundance of the latter was negatively related to rainfall. The overall prevalence of A. cantonensis in B. similaris was 24.6%. A. fulica was the most important intermediary host of A. cantonensis in the studied area and B. similaris was secondary in importance for A. cantonensis transmission dynamics.

Angiostrongylus cantonensis infection in molluscs in the municipality of São Gonçalo, a metropolitan area of Rio de Janeiro, Brazil: role of the invasive species Achatina fulica in parasite transmission dynamics

The aim of this study was to analyse the infection dynamics ofAngiostrongylus cantonensisin its possible intermediate hosts over two years in an urban area in the state of Rio de Janeiro where the presence ofA. cantonensis had been previously recorded in molluscs. Four of the seven mollusc species found in the study were exotic.Bradybaena similariswas the most abundant, followed byAchatina fulica, Streptaxissp., Subulina octona, Bulimulus tenuissimus, Sarasinula linguaeformisand Leptinaria unilamellata. Only A. fulicaand B. similariswere parasitised by A. cantonensis and both presented co-infection with other helminths. The prevalence of A. cantonensisin A. fulicawas more than 50% throughout the study. There was an inverse correlation between the population size ofA. fulicaand the prevalence of A. cantonensisand abundance of the latter was negatively related to rainfall. The overall prevalence of A. cantonensisin B. similariswas 24.6%. A. fulicawas the most important intermediary host of A. cantonensisin the studied area andB. similariswas secondary in importance for A. cantonensistransmission dynamics.

The Apple Snail Pomacea maculata (Caenogastropoda: Ampullariidae) as the intermediate host of Stomylotrema gratiosus (Trematoda: Stomylotrematidae) in Brazil: the first report of a mollusc host of a Stomylotrematid Trematode.

Trematodes belonging to the family Stomylotrematidae are intestinal parasites of birds. Despite the worldwide distribution and diversity of host species, the first intermediate host remains unknown. For a survey of parasites of Pomacea maculata , snails were collected from the municipality of São Vicente Férrer, state of Maranhão, northeastern Brazil. In the present study, the xiphidiocercariae shed from these snails were used in the experimental infection of the water bug Belostoma plebejum. The insect mortality was observed 30 days post-infection, and the metacercariae recovered in the body cavity of B. plebejum were identified as Stomylotrema gratiosus. This is the first report of an ampullariid snail as intermediate host of stomylotrematid trematodes.

Workshop on research priorities for management and treatment of angiostrongyliasis(1).

In a concluding session of the workshop, the participants developed a list of 115 research and outreach needs, outlining the top 5-7 needs in each of 8 areas (Table). For complete information, including presenter details and abstracts, visit the workshop website at www.hawaii.edu/cowielab/Angio%20website%20home.htm.

Phylogenetic relationship of the Brazilian isolates of the rat lungworm Angiostrongylus cantonensis (Nematoda: Metastrongylidae) employing mitochondrial COI gene sequence data.

BACKGROUND: The rat lungworm Angiostrongylus cantonensis can cause eosinophilic meningoencephalitis in humans. This nematode's main definitive hosts are rodents and its intermediate hosts are snails. This parasite was first described in China and currently is dispersed across several Pacific islands, Asia, Australia, Africa, some Caribbean islands and most recently in the Americas. Here, we report the genetic variability among A. cantonensis isolates from different geographical locations in Brazil using mitochondrial cytochrome c oxidase subunit I (COI) gene sequences. METHODS: The isolates of A. cantonensis were obtained from distinct geographical locations of Brazil. Genomic DNAs were extracted, amplified by polymerase reaction, purified and sequenced. A partial sequence of COI gene was determined to assess their phylogenetic relationship. RESULTS: The sequences of A. cantonensis were monophyletic. We identified a distinct clade that included all isolates of A. cantonensis from Brazil and Asia based on eight distinct haplotypes (ac1, ac2, ac3, ac4, ac5, ac6, ac7 and ac8) from a previous study. Interestingly, the Brazilian haplotype ac5 is clustered with isolates from Japan, and the Brazilian haplotype ac8 from Rio de Janeiro, São Paulo, Pará and Pernambuco states formed a distinct clade. There is a divergent Brazilian haplotype, which we named ac9, closely related to Chinese haplotype ac6 and Japanese haplotype ac7. CONCLUSION: The genetic variation observed among Brazilian isolates supports the hypothesis that the appearance of A. cantonensis in Brazil is likely a result of multiple introductions of parasite-carrying rats, transported on ships due to active commerce with Africa and Asia during the European colonization period. The rapid spread of the intermediate host, Achatina fulica, also seems to have contributed to the dispersion of this parasite and the infection of the definitive host in different Brazilian regions.

First record of a nematode Metastrongyloidea (Aelurostrongylus abstrusus larvae) in Achatina (Lissachatina) fulica (Mollusca, Achatinidae) in Brazil.

Achatina (Lissachatina) fulica was introduced in Brazil in the 1980s for commercial purposes ("escargot" farming) and nowadays, mainly by human activity, it is widespread in at least 23 out of 26 Brazilian states and Brasília, including the Amazonian region and natural reserves, where besides a general nuisance for people it is a pest and also a public health concern, since it is one of the natural intermediate host of Angiostrongylus cantonensis, ethiological agent of the meningoencephalitis in Asia. As Brazil is experiencing the explosive phase of the invasion, the Laboratório de Malacologia do Instituto Oswaldo Cruz/Fiocruz has been receiving samples of these molluscs for identification and search for Angiostrongylus cantonensis and Angiostrongylus costaricensis larvae. While examining samples of A. fulica different nematode larvae were obtained, including Aelurostrongylus, whose different species are parasites of felids, dogs, primates, and badger. Morphological and morphometric analyses presented herein indicated the species Aelurostrongylus abstrusus, as well as the occurrence of other nematode larvae (Strongyluris-like) found in the interior of the pallial cavity of A. fulica. This is the first report in Brazil of the development of A. abstrusus infective larvae in A. fulica evidencing the veterinary importance of this mollusc in the transmission of A. abstrusus to domestic cats. Since the spread of A. fulica is pointed out in the literature as one of the main causative spread of the meningoencephalitis caused by A. cantonensis the authors emphasize the need of sanitary vigilance of snails and rats from vulnerable areas for A. cantonensis introduction as the port side areas.

Freshwater snails and schistosomiasis mansoni in the state of Rio de Janeiro, Brazil: VI - Noroeste Fluminense Mesoregion

In this paper, the last of a series dealing with the survey of freshwater gastropods of the state of Rio de Janeiro, the results of collections carried out in the Noroeste Fluminense Mesoregion from 2002 to 2005 are presented and revealed the occurrence of 20 species: Antillorbis nordestensis; Biomphalaria glabrata; B. straminea; B. tenagophila; Drepanotrema anatinum; D. cimex; D. depressissimum; D. lucidum; Ferrissia sp.; Gundlachia ticaga; Gundlachia sp.; Heleobia sp.; Idiopyrgus sp.; Lymnaea columella; Melanoides tuberculatus; Physa acuta; P. marmorata; Plesiophysa guadeloupensis; Pomacea lineata; and Pomacea sp. Concerning the snail hosts of schistosomiasis the three natural vectors were identified and, although no specimens were found harbouring larval forms of Schistosoma mansoni, different kinds of cercariae had been observed.

Freshwater snails and schistosomiasis mansoni in the state of Rio de Janeiro, Brazil: VI--Noroeste Fluminense Mesoregion.

In this paper, the last of a series dealing with the survey of freshwater gastropods of the state of Rio de Janeiro, the results of collections carried out in the Noroeste Fluminense Mesoregion from 2002 to 2005 are presented and revealed the occurrence of 20 species: Antillorbis nordestensis; Biomphalaria glabrata; B. straminea; B. tenagophila; Drepanotrema anatinum; D. cimex; D. depressissimum; D. lucidum; Ferrissia sp.; Gundlachia ticaga; Gundlachia sp.; Heleobia sp.; Idiopyrgus sp.; Lymnaea columella; Melanoides tuberculatus; Physa acuta; P. marmorata; Plesiophysa guadeloupensis; Pomacea lineata; and Pomacea sp. Concerning the snail hosts of schistosomiasis the three natural vectors were identified and, although no specimens were found harbouring larval forms of Schistosoma mansoni, different kinds of cercariae had been observed.

Freshwater snails and schistosomiasis mansoni in the state of Rio de Janeiro, Brazil: V -- Norte Fluminense Mesoregion.

In this paper, the fifth of a series dealing with the survey of freshwater gastropods of the state of Rio de Janeiro, the results of collections carried out in the Norte Fluminense Mesoregion from 2002 to 2003 are presented and revealed the occurrence of 19 species: Antillorbis nordestensis; Burnupia sp.; Biomphalaria tenagophila; Drepanotrema anatinum; Drepanotrema cimex; Drepanotrema depressissimum; Drepanotrema lucidum; Ferrissia sp.; Gundlachia ticaga; Gundlachia sp.; Heleobia sp.; Hebetancylus moricandi; Idiopyrgus sp.; Lymnaea columella; Melanoides tuberculatus; Physa acuta; Physa marmorata; Pomacea sordida, and Pomacea sp. Concerning the snail hosts of Schistosoma mansoni only B. tenagophila was found, in contrast with other previuosly studied mesoregions.No specimens were found harbouring larval forms of S. mansoni although different kinds of cercariae had been observed. An account about the current schistosomiasis transmission sites in this Mesoregion is presented as well.