Raquel de Oliveira Simões

The giant African snail Achatina fulica as natural intermediate host of Angiostrongylus cantonensis in Pernambuco, northeast Brazil

The human cases of eosinophilic meningitis recently reported from Brazil have focused the attention of the public health agencies on the role the introduced snail Achatina fulica plays as hosts of the metastrongylid nematodes. Determining the potential of this snail to host and develop infective larval stages of metastrongylids in the wild and identify the species harbored by them is crucial for designing effective control measures. Here we assess if A. fulica may act as intermediate host of A. cantonensis at the peridomiciliary areas of a patients house from state of Pernambuco (PE), who was diagnosed with eosinophilic meningitis and a history of ingesting raw molluscs. Larvae obtained from naturally infected A. fulica were orally administered to Rattus norvegicus. The worms were collected from the pulmonary artery and brain, and were morphologically characterized and compared to the Japan isolate of A. cantonensis. Adult worms and infective L(3) larvae (PE isolate) recovered from A. fulica specimens were also analyzed by polymerase chain reaction and restriction fragment length polymorphism of ITS2 region from rDNA and compared to A. cantonensis (ES isolate), A. vasorum (MG isolate) and A. costaricensis (RS isolate). The large size of the spicules (greater than those observed in other species of Angiostrongylus) and the pattern of the bursal rays agree with the original species description by Chen (1935). Furthermore, the morphology of the PE isolate was similar to that of Japan isolate. The PCR-RFLP profiles obtained were distinctive among species and no variation in patterns was detected among adult individuals from A. cantonensis isolates from PE and ES. The importance of A. fulica as an intermediate host of eosinophilic menigoencepahlitis in Brazil is emphasized.

First report of Angiostrongylus cantonensis (Nematoda: Metastrongylidae) in Achatina fulica (Mollusca: Gastropoda) from Southeast and South Brazil

The rat lungworm Angiostrongylus cantonensis is a worldwide-distributed zoonotic nematode that can cause human eosinophilic meningoencephalitis. Here, for the first time, we report the isolation of A. cantonensis from Achatina fulica from two Brazilian states: Rio de Janeiro (specifically the municipalities of Barra do Piraí, situated at the Paraiba River Valley region and São Gonçalo, situated at the edge of Guanabara Bay) and Santa Catarina (in municipality of Joinville). The lungworms were identified by comparing morphological and morphometrical data obtained from adult worms to values obtained from experimental infections of A. cantonensis from Pernambuco, Brazil, and Akita, Japan. Only a few minor morphological differences that were determined to represent intra-specific variation were observed. This report of A. cantonensis in South and Southeast Brazil, together with the recent report of the zoonosis and parasite-infected molluscs in Northeast Brazil, provide evidence of the wide distribution of A. cantonensis in the country. The need for efforts to better understand the role of A. fulica in the transmission of meningoencephalitis in Brazil and the surveillance of molluscs and rodents, particularly in ports, is emphasized.

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

BackgroundThe 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.MethodsThe 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.ResultsThe 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.ConclusionThe 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.

Endemic angiostrongyliasis in the Brazilian Amazon: Natural parasitism of Angiostrongylus cantonensis in Rattus rattus and R. norvegicus, and sympatric giant African land snails, Achatina fulica

Angiostrongylus cantonensis, the rat lungworm, is one etiological agent of eosinophilic meningoencephalitis in humans. This zoonosis is frequently found in Asia and, more recently, in North America, Caribbean Island and northeastern of South America. Until now, research of A. cantonensis in southern, southeastern and northeastern regions of Brazil has been found natural infections only terrestrial and freshwater intermediate snail hosts (Achatina fulica, Sarasinula marginata, Subulina octona, Bradybaena similaris and Pomacea lineate). In this study, we examined the occurrence of helminthes in the synantropic rodents Rattus rattus and Rattus norvegicus in northern Brazil, focusing on the role of these species as vertebrate hosts of A. cantonensis and A. fulica as intermediate host have found natural. Thirty specimens of R. rattus and twelve of R. norvegicus were collected in the Guamá and Jurunas neighborhoods of the city of Belém, in the Brazilian state of Pará, of which almost 10% harbored adult worms in their pulmonary arteries. Sympatric A. fulica were found to be infected by L(3) larvae, which experimental infection confirmed to be A. cantonensis. Natural infection of snails and rodents with A. cantonensis was confirmed through morphological and morphometrical analyses of adults and larvae using light microscopy, scanning electron microscopy and molecular sequences of partial Cytochrome Oxidase subunit I. Phylogenetic analyses showed that A. cantonensis isolated from Pará, Brazil is similar to Japan isolate; once these specimens produced a single haplotype with high bootstrap support with Rio de Janeiro isolate. This study confirms that A. cantonensis is now endemic in northern Brazil, and that R. rattus and R. norvegicus act as natural definitive hosts, and A. fulica as the intermediate host of the parasite in this region.

Angiostrongyliasis in the Americas

The helminth Angiostrongylus costaricensis Morera & Cespedes, 1971 is endemic to the Americas and is responsible for a pathological abdominal syndrome, caused by the presence of the adult helminth in the mesenteric arteries. This is microscopically characterized by eosinophilic infiltration, vascular abnormality and a granulomatous reaction (GraeffTeixeira et al., 1987). The disease was first reported in humans by Cespedes et al. (1967) and Morera (1967) in Costa Rica. The adult worm was subsequently described from specimens recovered during surgical procedures by Morera & Cespedes (1971). The parasite’s current distribution ranges from the southern United States to northern Argentina (Morera, 1988), with human cases having been reported in Costa Rica, Honduras, Colombia, Martinique, Dominican Republic, Puerto Rico, Nicaragua, Mexico, Venezuela, Guadalupe, El Salvador, Panama and Brazil (Kaminsky, 1995).

A New Metastrongilid Species (Nematoda: Metastrongylidae): A Lungworm from Akodon montensis (Rodentia: Sigmodontinae) in Brazil

Abstract Angiostrongylus lenzii n. sp. (Nematoda: Metastrongyloidea) is described and illustrated from the pulmonary artery of the wild rodent Akodon montensis Thomas, 1913, collected in the municipality of Teresopólis, Rio de Janeiro State, Brazil. It represents the 17th species of the genus and the 2nd in South America as parasites in rodents. This new species is the most similar morphologically to Angiostrongylus vasorum, Angiostrongylus gubernaculatus, Angiostrongylus schmidti, and Angiostrongylus morerai. It can be distinguished from them by the morphology of the caudal bursa, ventral rays 2 and 3 with conspicuous knobs, differences in width and length of lateral and dorsal rays, presence of rays 8, and length of spicules.

Biochemical and histological responses of Rattus novergicus (Wistar) infected by Echinostoma paraensei (Trematoda: Echinostomatidae)

Tests were performed to evaluate the biochemical alterations in Rattus norvegicus after infection by the intestinal trematode Echinostoma paraensei. The rodents received 150 metacercariae each, serum samples were collected and the parasite load was quantified weekly until the fifth week of infection. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALKP), gamma-glutamyl transferase (GGT), bilirubin, glucose, total proteins and fractions and hepatic glycogen were determined. All the animals exposed to the metacercariae were infected in the first week and worms were recovered up to the third week after infection. The levels of AST, ALT, GGT, bilirubin and globulin rose in the first and/or second week and declined thereafter to levels near those of the control group. In contrast, the level of total proteins in the plasma fell significantly in the first week while the ALKP activity went down only in the fourth and fifth weeks in relation to the control group. There was no significant difference in the levels of albumin, glycogen and glucose. Infection by E. paraensei in R. norvegicus causes changes in the hepatic function, possibly resulting from the cholestasis produced by the partial obstruction of the bile duct by the helminths.

A New Heligmonellid Species (Nematoda) from Oligoryzomys nigripes (Rodentia: Sigmodontinae) in the Atlantic Forest, Brazil

Abstract Stilestrongylus lanfrediae n. sp. is described from the small intestine of Oligoryzomys nigripes (Cricetidae: Sigmodontinae) collected in the Atlantic Forest (Rio de Janeiro State, Teresópolis, Brazil). The new species shows some similarities to Stilestrongylus stilesi, Stilestrongylus freitasi, Stilestrongylus inexpectatus, Stilestrongylus moreli, and Stilestrongylus andalgala, but it can be distinguished from these species by the following combination of characters: 26 ridges in males and 25 in females at the mid-body, asymmetrical caudal bursa with a pattern of type 2-2-1, rays 6 markedly shorter than other lateral rays, rays 8 inserted asymmetrically on dorsal trunk and shorter than other species, and a proportion of spicule length in relation to body length (SpL/BL) of 25–29%. The new species is also distinguished from other species of Stilestrongylus by the asymmetry of the branches of the dorsal ray and by having the longest spicules.

Haematological alterations in Rattus norvegicus (Wistar) experimentally infected with Echinostoma paraensei (Trematoda: Echinostomatidae)

Laboratory rats (Rattus norvegicus) were infected with Echinostoma paraensei (Trematoda: Echinostomatidae). The rodents received 150 metacercariae each and blood samples were collected weekly until the fifth week of infection. The blood samples were analyzed for determination of haematocrit, total red blood cells with their dimensions, haemoglobin and haematimetric index (mean corpuscular volume, MCV; mean corpuscular haemoglobin, MCH; and mean corpuscular haemoglobin concentration, MCHC) and platelets. Red blood cells, haematocrit and haemoglobin in the first week had significantly lower levels than those of uninfected (control) rats, suggesting the development of normocytic and normocromic anaemia with anisocytic alteration. The number of eosinophils did not increase significantly among the groups. We concluded that E. paraensei produces haematological alterations in R. norvegicus, causing regenerative anaemia. This system can therefore be a useful model to study the direct and indirect effects of gastrointestinal infections.

A new heligmonellid (Nematoda: Heligmonellidae) from Oecomys mamorae (Rodentia: Sigmodontinae) in the Pantanal and new data on the synlophe of Guerrerostrongylus zetta from the Atlantic Forest, Brazil

Abstract: A new species of Heligmonellidae nematode, Guerrerostrongylus gomesae n. sp., is described from specimens collected from the small intestine of the rodent Oecomys mamorae Thomas, 1906 in the Brazilian Pantanal. It differs from the 2 other species of the genus by the number of ridges in the synlophe 35–46 and 40–48 at the midbody in the male and female, respectively, by rays 8 arising at midlength of the dorsal trunk, ending near the margin of the caudal bursa, and the dorsal ray divided at the second–third part into 2 branches, each branch divided into 2 subequal subbranches. In addition, specimens of Guerrerostrongylus zetta Travassos, 1937 collected in Oligoryzomys nigripes Olfers, 1818 contain synlophe with 36–42 cuticular ridges in males and 38–42 in females at the midbody. By the characteristics of the synlophe and caudal bursa, Gerrerostrongylus gomesae n. sp. is considered a new species. We suggest that the number of cuticular ridges for this genus broadens the range of the cuticular ridges to at least 35 at midbody.