Haem uptake is essential for egg production in the haematophagous blood fluke of humans, Schistosoma mansoni

Schistosomes ingest host erythrocytes, liberating large quantities of haem. Despite its toxicity, haem is an essential factor for numerous biological reactions, and may be an important iron source for these helminths. We used a fluorescence haem analogue, palladium mesoporphyrin, to investigate pathways of haem acquisition, and showed that palladium mesoporphyrin accumulates in the vitellaria (eggshell precursor glands) and ovary of female Schistosoma mansoni. Furthermore, incubation of adult females in 10-100 μm cyclosporin A (IC50 = 2.3 μm) inhibits the uptake of palladium mesoporphyrin to these tissues, with tenfold reductions in fluorescence intensity of the ovary. In vitro exposure to cyclosporin A resulted in significant perturbation of egg production, reducing egg output from 34 eggs per female to 5.7 eggs per female over the incubation period, and retardation of egg development. We characterized a S. mansoni homologue of the haem-responsive genes of Caenorhabditis elegans. The gene (Smhrg-1) encodes a protein with a molecular weight of approximately 17 kDa. SmHRG-1 was able to rescue growth in haem transport-deficient HEM1Δ yeast. Transcriptional suppression of Smhrg-1 in adult S. mansoni worms resulted in significant delay in egg maturation, with 47% of eggs from transcriptionally suppressed worms being identified as immature compared with only 27% of eggs laid by control worms treated with firefly luciferase. Our findings indicate the presence of transmembrane haem transporters in schistosomes, with a high abundance of these molecules being present in tissues involved in oogenesis.

Characterisation of a secretory serine protease inhibitor (SjB6) from Schistosoma japonicum

BACKGROUND: Proteins belonging to the serine protease inhibitor (serpin) superfamily play essential physiological roles in many organisms. In pathogens, serpins are thought to have evolved specifically to limit host immune responses by interfering with the host immune-stimulatory signals. Serpins are less well characterised in parasitic helminths, although some are thought to be involved in mechanisms associated with host immune modulation. In this study, we cloned and partially characterised a secretory serpin from Schistosoma japonicum termed SjB6, these findings provide the basis for possible functional roles.

METHODS: SjB6 gene was identified through database mining of our previously published microarray data, cloned and detailed sequence and structural analysis and comparative modelling carried out using various bioinformatics and proteomics tools. Gene transcriptional profiling was determined by real-time PCR and the expression of native protein determined by immunoblotting. An immunological profile of the recombinant protein produced in insect cells was determined by ELISA.

RESULTS: SjB6 contains an open reading frame of 1160 base pairs that encodes a protein of 387 amino acid residues. Detailed sequence analysis, comparative modelling and structural-based alignment revealed that SjB6 contains the essential structural motifs and consensus secondary structures typical of inhibitory serpins. The presence of an N-terminal signal sequence indicated that SjB6 is a secretory protein. Real-time data indicated that SjB6 is expressed exclusively in the intra-mammalian stage of the parasite life cycle with its highest expression levels in the egg stage (p < 0.0001). The native protein is approximately 60 kDa in size and recombinant SjB6 (rSjB6) was recognised strongly by sera from rats experimentally infected with S. japonicum.

CONCLUSIONS: The significantly high expression of SjB6 in schistosome eggs, when compared to other life cycle stages, suggests a possible association with disease pathology, while the strong reactivity of sera from experimentally infected rats against rSjB6 suggests that native SjB6 is released into host tissue and induces an immune response. This study presents a comprehensive demonstration of sequence and structural-based analysis of a secretory serpin from a trematode and suggests SjB6 may be associated with important functional roles in S. japonicum, particularly in parasite modulation of the host microenvironment.

Functional characterization of SjB10, an intracellular serpin from Schistosoma japonicum

Serine protease inhibitors (serpin) play essential roles in many organisms. Mammalian serpins regulate the blood coagulation, fibrinolysis, inflammation and complement activation pathways. In parasitic helminths, serpins are less well characterized, but may also be involved in evasion of the host immune response. In this study, a Schistosoma japonicum serpin (SjB10), containing a 1212 bp open reading frame (ORF), was cloned, expressed and functionally characterized. Sequence analysis, comparative modelling and structural-based alignment revealed that SjB10 contains the essential structural motifs and consensus secondary structures of inhibitory serpins. Transcriptional profiling demonstrated that SjB10 is expressed in adult males, schistosomula and eggs but particularly in the cercariae, suggesting a possible role in cercarial penetration of mammalian host skin. Recombinant SjB10 (rSjB10) inhibited pancreatic elastase (PE) in a dose-dependent manner. rSjB10 was recognized strongly by experimentally infected rat sera indicating that native SjB10 is released into host tissue and induces an immune response. By immunochemistry, SjB10 localized in the S. japonicum adult foregut and extra-embryonic layer of the egg. This study provides a comprehensive demonstration of sequence and structural-based analysis of a functional S. japonicum serpin. Furthermore, our findings suggest that SjB10 may be associated with important functional roles in S. japonicum particularly in host-parasite interactions.

Communities of parasite metazoans in Piaractus brachypomus (Pisces, Serrasalmidae) in the lower Amazon River (Brazil).

The aim of this study was to investigate the component community of parasite metazoans of Piaractus brachypomus in the lower Amazon River, northern Brazil. From 34 necropsied fish, 27,384 metazoan parasites were collected, such as Anacanthorus spathulatus, Mymarothecium viatorum and Notozothecium janauachensis (Monogenoidea); Spectatus spectatus and Contracaecum sp (Nematoda); Clinostomum marginatum and Dadaytrema oxycephala (Digenea); and Argulus carteri and Ergasilus sp. (Crustacea). The dominant species was S. spectatus followed by monogenoidean species, and there was aggregated dispersion of parasites, except for D. oxycephala and Contracaecum sp., which presented random dispersion. Positive correlation among the abundance of the three monogenoideans species were found, thus indicating that there was no competition between the species of these parasites on the gills of hosts. The abundances of some parasite species showed positive correlations with the size of the hosts, but the condition factor of the fish was not affected by the parasitism levels. It showed that this host had a metazoan community characterized by high species richness of metazoans, low evenness and high diversity of parasites, with prevalence of endoparasites, including larval stages. This was the first record of C. marginatum, A. carteri, Ergasilus sp. and Contracaecum sp. for P. brachypomus.

Parasite communities of the predatory fish, Acestrorhynchus falcatus and Acestrorhynchus falcirostris, living in sympatry in Brazilian Amazon.

This study investigated the parasite communities of wild Acestrorhynchus falcatus and Acestrorhynchus falcirostris populations living in sympatry in Brazilian Amazon. In these two hosts, a total of 12 parasite species e 1-9 parasite species were found per fish, and 10 of these species are metazoans. Eight species of parasites were common to both host species and four of them exhibited differences in abundance and/or prevalence. Parasite communities of the hosts were taxonomically similar (83%) and composed of both ectoparasites and endoparasites, and characterized by high prevalence and high abundance of endoparasites and an aggregated dispersion pattern. For A. falcirostris, the dominant parasite was Ichthyophthirius multifiliis, and for A. falcatus, it was Piscinoodinium pillulare. Shannon diversity and Berger-Parker dominance were similar for both hosts, while the parasites species richness and evenness showed differences influenced by the ectoparasites species. These two populations of hosts that inhabited the same geographical area had different sizes, but were exposed to the same infective stages, and acquired qualitatively and quantitatively similar endoparasites community, thus indicating that the amounts and types of prey congeneric that they were eating were similar. Therefore, the overlap in the same occurrence area play an important role in the parasite communities to these phylogenetically related hosts.

Desvendando a resposta dos eosinófilos, dos neutrófilos e dos monócitos nas parasitoses intestinais, na asma e na associação de ambas em crianças

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Medicina, Pós-graduação em Medicina Tropical, 2015.

Eficácia de oxfendazol em nematóides isolados em ovinos no estado do Ceará.

2015

Detecção de resistência aos benzimidazóis em nematoides gastrintestinais no Ceará.

2015

A study on monogenean parasites from the gills of some cichlids(Pisces: Cichlidae) from the Brazilian Amazon.

The aim of this study was to investigate monogenean fauna in gills of Geophagus camopiensis, Pterophyllum scalare, Satanoperca jurupari,and Satanoperca acuticeps in a tributary from the Amazon River system in Brazil. A total of 2,148 monogenean specimens were collected from140 fish examined from March 2012 to March 2013, and 84.3% of these fish were parasitized by 1 or more species. Such monogeneans were:Sciadicleithrum geophagi, Sciadicleithrum juruparii, Gussevia spiralocirra and Gyrodactylus sp. However, only G. camopiensis was parasitizedby more than 1 species of monogenean, while S. jurupari and S. acuticeps were parasitized by the same species. Prevalence, mean intensity andmean abundance varied among host species and the highest levels of infection were by G. spiralocirra followed by S. geophagi, both parasiteswith aggregated dispersion. Abundance of monogeneans was not influenced by the size of the host. In G. camopiensis, the infection levels byS. geophagi did not vary during the rainy or drainage seasons. This is the first study on monogenean infections for G. camopiensis and S. acuticeps.