Nosema bombycis, a pathogen of silkworm pebrine, is an obligate unicellular eukaryotic parasite. It is reported that the spore wall proteins have essential functions in the adherence and infection process of microsporidia. To date, the information related to spore wall proteins from microsporidia is still limited. Here, a 44 kDa spore wall protein NbSWP16 was characterized in N. bombycis. In NbSWP16, a 25 amino acids signal peptide and 3 heparin binding motifs were predicted. Interestingly, a region that contains 3 proline-rich tandem repeats lacking homology to any known protein was also present in this protein. The immunofluorescence analysis (IFA) demonstrated that distinct fluorescent signals were detected both on the surface of mature spores and the germinated spore coats. Immunolocation by electron microscopy revealed that NbSWP16 localized on the exospore regions. Finally, spore adherence analysis indicated that spore adherence to host cell was decreased more than 20% by anti-NbSWP16 blocking compared with the negative control in vitro. In contrast with anti-NbSWP16, no remarkable decrement inhibition was detected when antibodies of NbSWP16 and NbSWP5 were used simultaneously. Collectively, these results suggest that NbSWP16 is a new exospore protein and probably be involved in spore adherence of N. bombycis.

Studies using antibodies to immunolocalize the Toxoplasma gondii dense granule protein GRA3, have shown that this protein associates strongly with the parasitophorous vacuole membrane (PVM). However, as there was no predicted membrane-spanning domain this highlighted an unanswered paradox. We demonstrate that the previously published sequence for GRA3 is actually an artificial chimera of 2 proteins. One protein, of molecular weight 65 kDa, shares the C-terminus with published GRA3 and possesses no significant sequence similarity with any protein thus far deposited in Genbank. The second, with a predicted molecular weight of 24 kDa shares the N-terminal region, is recognized by the monoclonal antibody 2H11 known to react with the dense granules of T. gondii and is therefore the authentic GRA3. The corrected GRA3 has an N-terminal secretory signal sequence and a transmembrane domain consistent with its insertion into the PVM. Antibodies to recombinant GRA3 recognize a protein of 24 kDa in T. gondii excretory–secretory antigen preparations. The signal peptide is necessary and sufficient to target GFP to the dense granules and parasitophorous vacuole. A homologue was identified in Neospora caninum. Finally, GRA3 possesses a dilysine ‘KKXX’ endoplasmic reticulum (ER) retrieval motif that rationalizes its association with PVM and possibly the host cell ER.

The invasive blood stage of malaria parasites, merozoites, are complex entities specialized for the capture and entry of red blood cells. Their potential for vaccination and other anti-malaria strategies have attracted much research attention over the last 40 years, and there is now a considerable body of data relating to their biology. In this article some of the major advances over this period and remaining challenges are reviewed.

Haemonchus contortus is one of the most pathogenic parasites of small ruminants (e.g. sheep and goat). The treatment of haemonchosis is complicated because of recurrent resistance of H. contortus to common anthelmintics. The aim of this study was to compare the metabolism of the anthelmintic drug flubendazole (FLU) and the activities of selected biotransformation enzymes towards model xenobiotics in 4 different strains of H. contortus: the ISE strain (susceptible to common anthelmintics), ISE-S (resistant to ivermectin), the BR strain (resistant to benzimidazole anthelmintics) and the WR strain (resistant to all common anthelmintics). H. contortus adults were collected from the abomasums from experimentally infected lambs. The in vitro as well as ex vivo experiments were performed and analysed using HPLC with spectrofluorimetric and mass-spectrometric detection. In all H. contortus strains, 4 different FLU metabolites were detected: FLU with a reduced carbonyl group (FLU-R), glucose conjugate of FLU-R and 2 glucose conjugates of FLU. In the resistant strains, the ex vivo formation of all FLU metabolites was significantly higher than in the susceptible ISE strain. The multi-resistant WR strain formed approximately 5 times more conjugates of FLU than the susceptible ISE strain. The in vitro data also showed significant differences in FLU metabolism, in the activities of UDP-glucosyltransferase and several carbonyl-reducing enzymes between the susceptible and resistant H. contortus strains. The altered activities of certain detoxifying enzymes might protect the parasites against the toxic effect of the drugs as well as contribute to drug-resistance in these parasites.

Ultrastructural examination of the parasite-host interface of three tetraphyllidean cestodes of Raja naevus (sectioned attached to the intestine) indicates that while the strobilar tegument of each closely follows the typical cestode pattern that of the bothridia has several modifications probably associated with attachment. Microtriches similar to those of the strobilum are absent, although small spines on Echeneibothrium sp. and Phyllobothrium piriei and wart-like structures on Acanthobothrium quadripartitum bothridia are considered to represent the osmiophilic tips of specialized microtriches. The bothridia of the three species have no apparent function in the absorption of food but evidence suggests that the apical pad of the myzorhynchus of Echeneibothrium is specialized for the uptake of nutrients directly from the submucosa of the host intestine to which it is applied. Attachment and sensory roles by the myzorhynchus are also indicated. Echeneibothrium and P. piriei bothridia severely damage the host mucosal surface, while those of A. quadripartitum do little harm. This difference is probably associated with the size, mobility and mode of attachment of the cestodes. There are indications that the bothridia of Echeneibothrium upset the metabolism of mucosal cells in the attachment region. The tips of the strobilar microtriches of A. quadripartitum are shown to function in the attachment of the strobilum to the host mucosa, and in the protection of the microthrix proximal surfaces by a filtering action.

1. Crepidostomum farionis inhabits the gall bladder, as well as the intestine and pyloric caeca, of the trout and grayling.

2. The life history of Crepidostomum farionis has been worked out and is based on the similarity of organisation of the cercaria, encysted larval trematode and the adult.

3. The first intermediate hosts in the life history of this trematode are Pisidium amnicum (Müll.) and Sphaerium corneum (L.) though the latter is unusual.

4. The second intermediate host is the larva of the mayfly, Ephemera danica (Müll.).

5. There are two generations of rediae, the first gives rise to daughter rediae, which in turn produce cercariae.

6. The rediae are characterised by the absence of ambulatory processes and a functional intestine.

7. The cercaria (n.sp.) possesses “eye spots,” stylet and gland cells (salivary?), and the excretory vesicle is tube-shaped.

8. The excretory system of the redia and the cercaria has been worked out in detail.

9. The relation of the parasites to their respective hosts is discussed. On account of the need for further observations definite conclusions are held over for a later paper.

1. The pseudosuckers and holdfast organ of the metacercaria and adult stages of Diplostomum phoxini do not secrete acid mucopolysaccharide.

2. Alkaline phosphatase is present in the holdfast organ and in the cuticle lining the holdfast organ but not in the forebody glands or pseudosuckers.

3. Non-specific esterase has been shown to be present in the metacercaria and adult, especially in the pseudosuckers, the forebody glands which supply the pseudosuckers and in the holdfast organ. There is very little non-specific esterase in the posterior lobe of the adult.

4. Results of tests with enzyme inhibitors have shown that there are probably both A-esterase and B-esterase in the pseudosuckers, forebody glands and holdfast organ and that there is a cholinesterase present in the nervous system.

5. Cystine is present in most of the cuticle of the metacercaria but is absent from the cuticle lining the holdfast organ and the pseudosuckers.

6. The pseudosuckers and forebody glands as well as the holdfast organ appear to play a part in extracorporeal digestion.

Although the folate metabolic pathway in malaria parasites is a major chemotherapeutic target, resistance to currently available antifolate drugs is an increasing problem. This pathway, however, includes a number of enzymes that, to date, have not been characterized despite their potential for clinical exploitation. As a step towards evaluation of additional targets in this pathway, we report the isolation and characterization of 3 new genes that encode homologues of GTP cyclohydrolase I (GTP-CH), dihydrofolate synthase/folylpolyglutamate synthase (DHFS/FPGS) and serine hydroxymethyltransferase (SHMT). The genes encoding GTP-CH and SHMT are unambiguously assigned to chromosome 12, while that for DHFS/FPGS is tentatively assigned to chromosome 13. All 3 genes are expressed in blood-stage parasites, yielding transcripts of which only ca 60–70% is accounted for by coding sequence. All 3 of the proteins predicted to be encoded by these genes display sequence differences compared to the human host homologues that may be of functional significance. These data bring the complement of cloned genes that encode activities in the pathway to seven, leaving only the gene encoding dihydroneopterin aldolase (DHNA) to be identified in the route from GTP to folate synthesis and folate turnover in the thymidylate cycle.

This study aimed to identify putative quantitative trait loci (QTL) that significantly affect internal parasite resistance in a backcross sheep population. A Romney×Merino backcross (to Merino) flock was challenged in 3 separate infections with Trichostrongylus colubriformis (primary and secondary) and Haemonchus contortus (tertiary). Haematological parameters were measured and faecal worm egg counts (FWEC) were established to estimate parasite burden. QTL mapping was conducted for FWEC and for the changes in haematocrit following H. contortus challenge and in eosinophil numbers following T. colubriformis challenge. Animals were genotyped for 55 microsatellite markers on selected chromosomes 2, 3, 6, 11, 13, 15, 21, and 22. Four putative quantitative trait loci were found; these being for eosinophil change in the primary infection (OAR 21), for FWEC in the first infection and eosinophil change in the secondary infection (OAR 3) and for FWEC in the secondary infection (OAR 22). No significant quantitative trait loci were detected for FWEC or haematocrit change during the Haemonchus contortus infection. The position of the putative quantitative trait loci for eosinophil change on OAR 3 is consistent with other reports of parasite resistance quantitative trait loci, implying some commonality between studies.

Evolutionary and closer structural relationships are demonstrated by phylogenetic analysis, peptide prediction and molecular modelling between Solanum tuberosum apyrase, Schistosoma mansoni SmATPase 2 and Leishmania braziliensis NDPase. Specific protein domains are suggested to be potentially involved in the immune response, and also seem to be conserved during host and parasite co-evolution. Significant IgG antibody reactivity was observed in sera from patients with American cutaneous leishmaniasis (ACL) and schistosomiasis using potato apyrase as antigen in ELISA. S. mansoni adult worm or egg, L. braziliensis promastigote (Lb) and Trypanosoma cruzi epimastigote (EPI) have ATP diphosphohydrolases, and antigenic preparations of them were evaluated. In ACL patients, IgG seropositivity was about 43% and 90% for Lb and potato apyrase, respectively, while IgM was lower (<19%) for both. In schistosomiasis patients IgM (>40%) or IgG (100%) seropositivity for both soluble egg (SEA) and adult worm (SWAP) antigens was higher than that found for potato apyrase (IgM=10%; IgG=39%). In Chagas disease, IgG seropositivity for EPI and potato apyrase was 97% and 17%, respectively, while the IgM was low (3%) for both antigens. The study of the conserved domains from both parasite proteins and potato apyrase could lead to the development of new drug targets or molecular markers.

Schistosome infections in humans are characterized by the development of chronic disease and high re-infection rates after treatment due to the slow development of immunity. It appears that anti-schistosome antibodies are at least partially mediating protective mechanisms. Efforts to develop a vaccine based on immunization with surface-exposed or secreted larval or worm proteins are ongoing. Schistosomes also express a large number of glycans as part of their glycoprotein and glycolipid repertoire, and antibody responses to those glycans are mounted by the infected host. This observation raises the question if glycans might also form novel vaccine targets for immune intervention in schistosomiasis. This review summarizes current knowledge of antibody responses and immunity in experimental and natural infections with Schistosoma, the expression profiles of schistosome glycans (the glycome), and antibody responses to individual antigenic glycan motifs. Future directions to study anti-glycan responses in schistosomiasis in more detail in order to address more precisely the possible role of glycans in antibody-mediated immunity are discussed.

Background/Objective. Multiple factors affect schistosomiasis transmission in distributed meta-population systems including age, behaviour, and environment. The traditional approach to modelling macroparasite transmission often exploits the ‘mean worm burden’ (MWB) formulation for human hosts. However, typical worm distribution in humans is overdispersed, and classic models either ignore this characteristic or make ad hoc assumptions about its pattern (e.g., by assuming a negative binomial distribution). Such oversimplifications can give wrong predictions for the impact of control interventions. Methods. We propose a new modelling approach to macro-parasite transmission by stratifying human populations according to worm burden, and replacing MWB dynamics with that of ‘population strata’. We developed proper calibration procedures for such multi-component systems, based on typical epidemiological and demographic field data, and implemented them using Wolfram Mathematica. Results. Model programming and calibration proved to be straightforward. Our calibrated system provided good agreement with the individual level field data from the Msambweni region of eastern Kenya. Conclusion. The Stratified Worm Burden (SWB) approach offers many advantages, in that it accounts naturally for overdispersion and accommodates other important factors and measures of human infection and demographics. Future work will apply this model and methodology to evaluate innovative control intervention strategies, including expanded drug treatment programmes proposed by the World Health Organization and its partners.

Secretions were induced from second (invasive) stage juveniles (J2s) of the potato cyst nematode Globodera rostochiensis by exposing them to 5-methoxy-N,N-dimethyl tryptamine oxalate (DMT). Secretions were collected from J2s in sufficient quantity to allow direct analysis. Gel electrophoresis followed by monochromatic silver staining demonstrated the presence of at least 10 proteins. The presence of several enzymes, including superoxide dismutase and proteases, was demonstrated using Western blots and activity assays. Antisera raised against the secretions recognized bands on Western blots consistent in molecular mass with those identified on silver stained gels. The antisera recognized structures implicated in the production of secretions including the subventral gland cells and surface of J2s.

Eimeria flavescens and E. irresidua from the domestic rabbit are redescribed. The relatively smaller ovoidal oocysts of E. flavescens which measure on average 31·7 × 21·4 μm, possess a wide micropyle at the broad end. First-generation schizonts of this species develop deep in the glands of the lower small intestine. Merozoites migrate to the caecum and colon where second, third and fourth-generation schizonts develop in the superficial epithelium followed by the fifth-generation schizonts and gametocytes which form in the glands. In young Dutch rabbits E. flavescens is very pathogenic; low doses of oocysts produce a severe enteritis with high mortality and morbidity. The larger, broadly ellipsoidal oocysts of E. irresidua measure on average 38·4 × 23·2 μm and often possess a very small cryptic oocyst residuum. The endogenous stages develop in the small intestine only; first-generation schizonts in the glands and second-generation schizonts in the lamina propria whilst third and fourth-generation schizonts and gametocytes develop in the villous epithelium. E. irresidua is not pathogenic in young Dutch rabbits; even heavy infections produce only a transient pause in weight gain.

Acetylcholinesterase (AChE) is present in all stages of the life-cycle of schistosomes and is located in muscle and on the surface of the parasite. Metrifonate is a drug that inhibits AChE. We compared the AChEs from three schistosome species (Schistosoma mansoni, Schistosoma haematobium and Schistosoma bovis) that have different susceptibilities to metrifonate in vivo. Sensitivities to AChE inhibitors were similar. The subunits of AChE were 110 kDa and 76 kDa and the dominant molecular form of AChE was a G2 form in all three species. This was the major form on the tegument while additional molecular forms were associated with the internal tissues. Differences in relative amounts of AChE activity between these species were found in the adults but not in the schistosomula. At the adult stage the major difference between species lay in the relative amounts of AChE activity in their teguments. S. haematobium teguments carried 20 times and S. bovis 6·9 times the activity present on S. mansoni teguments. These quantitative differences associate with the relative sensitivities of these species to metrifonate.

In two recent papersm Sambon and Seligmann (1907, 1908) have recorded some observations on the intracellular parasites of snakes, and have described no less than ten new species. The authors, in discussing the life histories of these parasites, have made the startling discovery that, “the life history of the haemogregarines like that of the haemoprotozoa is divided into two cycles: a schizogonic or ‘vegetative’ cycle spent in the blood of vertebrates and characterised by asexual multiplication, and a sporogonic or sexual cycle spent in the digestive organs of blood-sucking invertebrates and characterised by sexual reproduction.” The authors then go on to speak quite familiarly of young merozoites, adult schizonts, adult sporonts, and so on.

A group of monoclonal antibodies (MAb) was used to immunochemically characterize Babesia bigemina surface components. Two surface reactive MAb by an IFAT test were subsequently shown to bind to epitopes on the external surface of the parasite plasma membrane as evidenced by immunoelectron microscopy. Parasite components with relative sizes of 68, 62, 60, 58, 56, 54, 51, 49, 48, 47, 43, 36 kDa were identified with the group of MAb in at least 6 geographically different B. bigemina isolates. The antigenic components were demonstrated to be species-specific and apparently predominant. Both polyclonal antibodies (immune bovine serum) and highly diluted MAb specifically reacted with such bands producing a strong signal in Western blots.

The DNA from a number of free-living and parasitic nematode species was examined to determine the genomic number and distribution of DNA sequences encoding two evolutionary conserved proteins; the major sperm protein (MSP) and nematode actin. Ascaris and Caenorhabditis MSP cDNA sequences and Ascaris genomic actin sequences were used to probe Southern blots of Eco RI and Hin d III digested nematode DNA. The number of MSP genes varied widely between the 1 MSP gene in Ascaris and the 60 MSP genes in Caenorhabditis. Filarial nematodes appeared to have 1—4 MSP genes while the plant and insect parasitic species showed from 5–12 MSP-hybridizing restriction fragments. Mammalian intestinal parasites showed between 1 and 13 bands hybridizing with the MSP probes. Blots probed to estimate the number of actin genes showed that, with the exception of Ascaris which contains more than 20 germ line sequences that encode actin, all of the nematodes tested had between 3 and 9 bands that hybridized to the Ascaris genomic actin probe. The possible use of highly conserved sequences such as MSP and actin to differentiate between nematode species in diagnostic and taxonomic studies is discussed.

The current taxonomy of parasites in the genus Sarcocystis is largely based on morphological characteristics as well as on host specificity and life-cycle structure. Recently, phylogenetic analyses of partial ribosomal RNA (rRNA) sequences provided support for paraphyly of Sarcocystis. We have tested the validity of this hypothesis by sequencing the complete 18S rRNA genes of Sarcocystis arieticanis, Sarcocystis gigantea and Sarcocystis tenella and comparing them with gene sequences derived from other taxa of the phylum Apicomplexa. The results obtained from this study do not reject the hypothesis of monophyly of Sarcocystis species, although the bootstrap data were inconclusive for some species.

This paper describes in vitro antibody dependent cytotoxicity against Trypanosoma cruzi epimastigotes by normal mouse splenic lymphocytes. Cytotoxicity was expressed as the percentage reduction in the number of motile parasites upon incubation with lymphocytes at 37 °C in a denned medium. Failure of the non-motile parasites to regain motility and their ensuring degeneration at 28 °C in liver infusion tryptose (LIT) medium confirmed loss of motility as a criterion of cytotoxicity. Incubation of T. cruzi at 37 °C for 18 h in a defined medium per se did not interfere with motility but was followed by a lag phase of the growth curve in LIT medium at 28 °C. The lag phase was prolonged for T. cruzi which had previously been incubated at 37 °C in the absence of cells.