Schistosoma mansoni cercariae in water were shown to possess a largely aerobic energy metabolism, the Krebs cycle being the main terminal of carbohydrate breakdown. A metabolic transition towards a more anaerobic breakdown of carbo-hydrate could be achieved by incubation conditions which also stimulated biological transformation. Incubation of cercariae in a simple salt medium containing 5 mM glucose induced such a metabolic transition: beside carbon dioxide large amounts of lactate and pyruvate were excreted. The results indicate that the production of pyruvate was coupled to electron transfer in the respiratory chain. Some aspects of this unusual pyruvate production are discussed. The observed change in the end-product pattern of carbohydrate breakdown is very rapid: most of the switch occurred within 2 h. Our results show that the metabolic transition was triggered by the biological transformation itself, or by the same event that induces the biological transformation. The metabolic and the biological changes proceeded synchronously.

Ten colostrum-deprived, coccidia-free lambs were reared artificially in indoor cages. At 4 weeks of age they were allocated to 5 groups of 2: Groups 1–3 received 104 105 and 106 sporulated oocysts of Eimeria erandallis/lamb respectively, each as a single inoculum. Group 4 lambs received daily doses increasing over a 4-week period. Groups 1–3 showed diarrhoea and weight loss from about day 14, followed by slow recovery from about day 30. The diarrhoea was accompanied by a wave of oocyst output and a reduction in serum albumin and plasma alkaline phosphatase activity. One lamb in group 4 died of an unrelated condition on day 15; the other showed no clinical effect of the oocyst inoculations and its oocyst output was related only to the first week or two of inoculation. Lambs in Group I showed no clinical effect or oocyst output following challenge with 106 oocysts on day 28. E. crandallis showed high pathogenicity and immunogenicity. Its pathogenicity was not closely related to infecting dose within the range 104–106.

Two new microsporidia, Anostracospora rigaudi n. g., n. sp., and Enterocytospora artemiae n. g., n. sp. infecting the intestinal epithelium of Artemia parthenogenetica Bowen and Sterling, 1978 and Artemia franciscana Kellogg, 1906 in southern France are described. Molecular analyses revealed the two species belong to a clade of microsporidian parasites that preferentially infect the intestinal epithelium of insect and crustacean hosts. These parasites are morphologically distinguishable from other gut microsporidia infecting Artemia. All life cycle stages have isolated nuclei. Fixed spores measure 1·3×0·7 μm with 5–6 polar tube coils for A. rigaudi and 1·2×0·9 μm with 4 polar tube coils for E. artemiae. Transmission of both species is horizontal, most likely through the ingestion of spores released with the faeces of infected hosts. The minute size of these species, together with their intestinal localization, makes their detection and identification difficult. We developed two species-specific molecular markers allowing each type of infection to be detected within 3–6 days post-inoculation. Using these markers, we show that the prevalence of these microsporidia ranges from 20% to 75% in natural populations. Hence, this study illustrates the usefulness of molecular approaches to study prevalent, but cryptic, infections involving microsporidian parasites of gut tissues.

A short historical account is given of the writings on flea larvae since the seventeenth century. The larvae have been known since Leeuwenhoek tried to breed them in 1683.

The life-history of Ceratophyllus wickhami is described. The fleas were bred in the laboratory on a grey squirrel. Three larval instars occur, and the life-history occupies, on an average, 6 weeks under experimental conditions.

The external anatomy of the third instar larva of C. wickhami is described. The mouth parts are generalised and suggest the condition of a primitive insect. The tracheal system is composed of a double longitudinal trunk on each side, with spiracles on the prothorax, metathorax and first eight abdominal segments.

The first and second instar of C. wickhami are similar to the third instar larvae, except in size and the presence of a hatching spine in the first stage.

The larva of Ceratophyllus fasciatus is mentioned. The importance of the species lies in the presence of completely separated galea and lacinia.

Larvae of Xenopsylla cheopis and X. astia are briefly described. The larvae of the two species are practically identical except for the shape of the mandibles.

To detect the expression of pro-fibrotic molecules, such as heat shock protein 47 (Hsp47), transforming growth factor-beta 1 (TGF-β1) and connective tissue growth factor (CTGF) in liver specimens, and analyse their correlations with the progression of schistosomal hepatic fibrosis, liver biopsy was performed in 42 chronic schistosomiasis (CS) patients, 16 chronic hepatitis B (CHB) patients and five healthy individuals (HI). Immunohistochemistry (IHC) analyses displayed that the expression of Hsp47, TGF-β1 and CTGF was increased in CS and CHB patients compared with HI. Using real-time PCR, the mRNA levels of Hsp47, TGF-β1 and CTGF were higher in CS patients compared with HI. In CS patients, the mRNA levels of these genes were correlated with the stage of fibrosis, and TGF-β1 mRNA expression was associated with the grade of inflammation. Additional analyses indicated that the mRNA levels of Hsp47 and CTGF were highly correlated with liver stiffness value and spleen thickness diameter, both of which represented the severity of fibrosis. In conclusion, the three molecules are involved in the pathogenesis of hepatic fibrosis infected by Schistosoma japonicum. TGF-β1 participates not only in the inflammatory process, but also in the fibrotic process in which Hsp47 and CTGF probably play a key role.

The genetic diversity of haematozoan parasites in island avifauna has only recently begun to be explored, despite the potential insight that these data can provide into the history of association between hosts and parasites and the possible threat posed to island endemics. We used mitochondrial DNA sequencing to characterize the diversity of 2 genera of vector-mediated parasites (Plasmodium and Haemoproteus) in avian blood samples from the western Indian Ocean region and explored their relationship with parasites from continental Africa. We detected infections in 68 out of 150 (45 3%) individuals and cytochrome b sequences identified 9 genetically distinct lineages of Plasmodium spp. and 7 lineages of Haemoproteus spp. We found considerable heterogeneity in parasite lineage composition across islands, although limited sampling may, in part, be responsible for perceived differences. Two lineages of Plasmodium spp. and 2 lineages of Haemoproteus spp. were shared by hosts in the Indian Ocean and also on mainland Africa, suggesting that these lineages may have arrived relatively recently. Polyphyly of island parasites indicated that these parasites were unlikely to constitute an endemic radiation and instead probably represent multiple colonization events. This study represents the first molecular survey of vector-mediated parasites in the western Indian Ocean, and has uncovered a diversity of parasites. Full understanding of parasite community composition and possible threats to endemic avian hosts will require comprehensive surveys across the avifauna of this region.

We have attempted to provide a rational basis for improving the protocols for chemotherapy of malaria. We model the regression of parasitaemia by Plasmodium falciparum, its subsequent elimination from the body, or recrudescence, for populations of cells treated with chloroquine. Our model assumes that the drug forms a complex with some receptor in the parasite and that parasites possessing this complex die at a defined rate. We take into account that chloroquine is eliminated exponentially from the body. We show how the parameters of the model can be derived from observations in the field. The model correctly predicts the effects of drug dose, degree of initial parasitaemia, rate of parasite multiplication and degree of drug resistance to chloroquine chemotherapy. The level of parasitaemia will reduce to a minimum at sufficiently high concentrations of chloroquine, but only if the parasitaemia is reduced to below that of 1 parasite per infected person will a cure of malaria be obtained. Otherwise, recrudescence will, sooner or later, occur. We show that, even for drug-resistant malaria, if 2 doses of chloroquine are given to a patient with an interval of some 10 days between them, parasites can be eliminated from the body without toxic levels of chloroquine being reached.

Amylopectin is used for carbohydrate storage in different life-stages of a number of apicomplexan parasites. We have performed an ultrastructural analysis of amylopectin granules from the oocyst residual body and sporozoites of Cryptosporidium parvum. Amylopectin granules were studied in situ and after isolation from ‘French’ press disrupted parasites, by conventional transmission electron microscopy (TEM) of sectioned oocysts and various negative staining and cryoelectron microscopy techniques. Within the membrane-enclosed oocyst residuum large amylopectin granules (0·1–0·3 μm) can be found besides a characteristic large lipid body and a crystalline protein inclusion. Smaller granules were detected in sectioned sporozoites. Negative staining of isolated amylopectin granules revealed some ultrastructural features not readily visible in sectioned material. The large amylopectin granules had a smooth surface with a ‘ball of string’-like inner structure. Granules isolated from sporozoites were more irregularly shaped and showed a rod-like particulate composition. With the exception of α-amylase, which led to some degree of damage of the surface of the particles, treatment of amylopectin granules with other glycohydrolases had little effect on the overall structure. However, granules adhered to one another. Only when the granules were boiled did the ‘ball of string’ structure gradually dissolve.

The Regent Honeyeater (Xanthomyza phyrigia) is an endangered Australian bird species. Breeding populations have been established at Australian zoos in support of re-introduction programmes. This species is the host of a new species of Isospora (Apicomplexa). Oocysts are spherical, 25·8 (22·5–28·75) by 23·8 (20–26·25) μm with a colourless to pale yellow smooth wall undergoing rapid exogenous sporulation, 90% sporulated oocysts in 8 h at 20°C. Each oocyst contains 1 polar granule. Sporocysts are ovoid, 18·67 (17–19) by 9·49 (9–10) μm with a flat Stieda body and spherical substieda body devoid of a hyaline body. The asexual stages and sexual phase is within the enterocytes of the duodenum and jejunum. Faeces collected in the morning (AM, n=84) and in the afternoon (PM, n=90) revealed significant diurnal periodicity in oocyst shedding; 21% (18 of 84) of the AM were positive with the mean of 499 oocysts.g−1 compared to the PM with 91% (82 of 90) bird faeces positive with the mean of 129 723 oocysts.g−1. Therefore, parasite checks for these birds should be carried out in the afternoon to obtain an accurate result. The ecological significance of the high parasite burden in captive birds requires further investigation and comparison to the wild counterparts.

The aims of the present work were to test the effect of tamoxifen administered topically and the therapeutic efficacy of tamoxifen and pentavalent antimonial combinations in an experimental model of cutaneous leishmaniasis. BALB/c mice infected with a luciferase expressing line of Leishmania amazonensis were treated with topical tamoxifen in two different formulations (ethanol or oil-free cream) as monotherapy or in co-administration with pentavalent antimonial. Treatment efficacy was evaluated by lesion size and parasite burden, quantified through luminescence, at the end of treatment and 4 weeks later. Topical tamoxifen, formulated in ethanol or as a cream, was shown to be effective. The interaction between tamoxifen and pentavalent antimonial was additive in vitro. Treatment with combined schemes containing tamoxifen and pentavalent antimonial was effective in reducing lesion size and parasite burden. Co-administration of tamoxifen and pentavalent antimonial was superior to monotherapy with antimonial.

Two new myxosporean species in the gallbladders of frogs have recently spread across eastern Australia and cause disease. Cystodiscus axonis sp. n. and Cystodiscus australis sp. n. are species of Myxosporea (Myxozoa) identified from a range of Australian frogs and tadpoles including the introduced Cane toad (Rhinella marina). The new species are defined by their distinct genetic lineage, myxospore morphology and ultrastructure of the pre-sporogonic development. Spores of both species are produced in the gallbladder. Spores of C. axonis sp. n. possess distinct filiform polar appendages (FPA). The pre-sporogonic development of C. axonis sp. n. is within myelinated axons in the central nervous system of hosts, as well as bile ducts of tadpoles. Pre-sporogonic and sporogonic development of C. australis sp. n. is confined to tadpole bile ducts and myxospores of C. australis sp. n. are devoid of FPA. The genus Cystodiscus Lutz, 1889 introduced for Cystodiscus immersus Lutz, 1889 is emended to accompany myxosporean parasites affecting amphibians previously classified in the genus Myxidium sensu lato. A synopsis of described species within Cystodiscus is provided.

The planorbid snail Biomphalaria glabrata responded to exposure to either the compatible digenetic trematode Echinostoma paraensei or the incompatible species Echinostoma trivolvis by producing increased amounts of several distinctive plasma polypeptides. These polypeptides characteristically precipitated from plasma when mixed with secreted–excreted products (SEP) of sporocysts or rediae from either digenean species. In contrast, control snails, or snails that had been wounded or infected with bacteria (Serratia marcesens or Staphylococcus epidermidis) showed no obvious plasma alterations and no precipitates formed when their plasma was mixed with SEP. Another planorbid species, Helisoma trivolvis, which displays reverse compatibility for the echinostome species used, also responded to exposure to both echinostomes by increased production of plasma polypeptides that precipitated in the presence of SEP. With some individual variation, these 2 snail species synthesized SEP-reactive plasma polypeptides forming diffuse bands centred at 53, 65, 80–120 and 200 kDa (the latter absent in Helisoma trivolvis). The 53 kDa polypeptides had not been observed before, whereas the others have been noted from B. glabrata. The diffuse 65 kDa band was strongly bound by anti-fibrinogen antibodies, supportive of earlier studies indicating it contains fibrinogen-related domains. The other specified polypeptides were also bound by these antibodies raising the possibility that they too contain fibrinogen domains. The results are suggestive of a general ability of these 2 planorbid snails to detect the presence of echinostomes even if the latter are subsequently incapable of development. The complex response they then mount, one not evoked by other challenges such as wounding or bacterial infection, may represent a dedicated response to a frequently encountered group of pathogenic parasites, the digeneans (echinostomes).

Chagas disease is a public health problem, affecting about 7 million people worldwide. Benznidazole (BZN) is the main treatment option, but it has limited effectiveness and can cause severe adverse effects. Drug delivery through nanoparticles has attracted the interest of the scientific community aiming to improve therapeutic options. The aim of this study was to evaluate the cytotoxicity of benznidazole-loaded calcium carbonate nanoparticles (BZN@CaCO3) on Trypanosoma cruzi strain Y. It was observed that BZN@CaCO3 was able to reduce the viability of epimastigote, trypomastigote and amastigote forms of T. cruzi with greater potency when compared with BZN. The amount of BZN necessary to obtain the same effect was up to 25 times smaller when loaded with CaCO3 nanoparticles. Also, it was observed that BZN@CaCO3 enhanced the selectivity index. Furthermore, the cell-death mechanism induced by both BZN and BZN@CaCO3 was evaluated, indicating that both substances caused necrosis and changed mitochondrial membrane potential.

Cattle were inoculated with purified Theileria parva piroplasm and/or schizont antigen. Two similar inoculations were given 10 days apart. Serum samples were regularly collected and the indirect haemagglutination (IHA), indirect immunofluorescence (IFA), complement-fixation (CF) and immunodiffusion (ID) tests were used to detect T. parva antibodies. Selected sera were separated by Sephadex G 200 and fractions examined for specific immunoglobulin activity.

With the IHA test specific antibody first appeared 4 days post inoculation (dpi) and reached high titres by 8 dpi. With the IFA and CF tests specific antibody also appeared 4 dpi and reached high titres by 8–14 and 10–14 dpi respectively. Specific ID activity was detected as early as 6 dpi and persisted for the length of the experiment.

On fractionation of the sera both the IHA and CF tests indicated a sequential production of T. parva IgM and 7S Ig. However, in cattle inoculated with schizont antigen only, a significant 7S Ig component was not detected by the IHA test until 16 dpi, but was demonstrated by the CF test 7 dpi. The IFA test, however, detected anti T. parva activity almost exclusively in the 7S Ig fractions. All cattle were challenged 35–42 dpi with infective T. parva stabilate and all cattle proved fully susceptible. The lack of protection by high titres of specific T. parva immunoglobulin is discussed, together with the data on the differential immunoglobulin response, in relation to previous results.

The presence of living tapeworm increases the rate of hydrolysis of starch by α-amylase in vitro. This effect indicates that ‘membrane digestion’ may be one function of the tegument of tapeworms. The effect varies with the surface area and region of the tapeworm. Fixed tapeworm pieces do not enhance starch hydrolysis. The results may provide evidence both for and against the current explanations of membrane digestion. Some possible mechanisms involved in membrane digestion in tapeworms are considered. The importance of membrane digestion in the physiology of gut parasites and the possibility of its wide occurrence are discussed.

We wish to thank Dr J. Llewellyn for his criticism and advice during the preparation of this manuscript and for providing rats and mice infected with Hymenolepis. We also thank Dr R. A. Thornhill for examining some tapeworm tissue under the electron microscope; also Mr A. Wilson of the Meat Inspection Department of the Birmingham Meat Market for his unfailing co-operation.

This study reports on morphological variability of Eimeria species, which may be given either by drawings or as quantitative data. The drawings may be used to facilitate identification by eye of ‘unknown’ Eimeria specimens, whereas quantitative data may serve as a reference set for identification by multivariate statistical techniques. The morphology of 810 Eimeria specimens was defined in binary (b/w) digital images by pixels of their oocyst outline. A Fourier transform of pixel positions yielded size and shape features. To classify coccidia, the quantitative data were employed in an agglomerative clustering by average linkage algorithm with equal weight assigned to size and shape. An inverse Fourier transform served to reconstruct oocyst outlines, i.e. outlines of average shape and size, from mean values of features in resulting clusters. Clusters were subsequently identified based on their average morphology by comparison with drawings of species in an earlier taxonomical work. Five hundred oocyst outlines were simulated for each cluster representing a species, and shape/size variability was presented in contour diagrams. Differences in species shapes, and correspondence in length and width, were seen after reconstruction by inverse Fourier transform and comparison with earlier studies.

Here we have studied phospholipase A1 (Plase A1) from Trypanosoma cruzi infective stages and it's possible role regarding the interaction with mammalian host cells. Plase A1 was mainly detected as a membrane-bound activity in the infective amastigote and trypomastigote stages, being remarkably higher with respect to the non-infective epimastigotes. It is noteworthy that only the infective stages secreted Plase A1. Moreover, along the differentiation process from epimastigotes into metacyclic trypomastigotes, the secreted enzyme activity increased simultaneously with the appearance of metacyclic forms, as expected. Since this enzyme is predominantly membrane-associated and secreted by the infective stages, Vero cell lipid profile modifications were analysed after interaction with either intact infective parasites or purified T. cruzi Plase A1. Significant changes in Vero cell lipid composition were observed, with the appearance of free fatty acids, diacylglycerol and lysophosphatidylcholine. Concomitantly with the generation of second lipid messengers, host cell protein kinase C activation was demonstrated. These results indicate that T. cruzi Plase A1 could play a critical role in the early events of parasite-host cell interaction that precede invasion.

A monoclonal antibody (OW-IGO) raised against purified excysted oocysts of Cryptosporidium parvum reacted in an immunofluorescence assay with the oocyst wall. The corresponding antigen was localized by immunoelectron microscopy in fibrillous material present in the parasitophorous vacuole of developing macrogametes and in the wall of both single and double layered sporulating oocysts. Gold particles were also detected over electron-lucent vesicles of the macrogametes by immunoelectron microscopy. On Western blotting of C. parvum oocyst extracts, major bands at 250 and 40 kDa and several minor components were recognized by Mab OW-IGO. Almost complete abolition of Western blot reactivity occurred after periodate oxidation of oocyst antigen, suggesting that monoclonal antibody OW-IGO reacts with a carbohydrate epitope. Taken together, our results suggest that a fibrillous glycoproteinic material is released in the parasitophorous vacuole from electron-lucent vesicles during gametogenesis, and later condensed in the oocyst wall.

Parasitism is often a source of variation in host's fitness components. Understanding and estimating its relative importance for fitness components of hosts is fundamental from physiological, ecological and evolutionary perspectives. Host-parasite studies have often reported parasite-induced reduction of host fecundity, whereas the effect of parasitism on host survival has been largely neglected. Here, we experimentally investigated the effect of infestation by rat fleas (Nosopsyllus fasciatus) on the life span of wild-derived male common voles (Microtus arvalis) bred in captivity. We found that the mean life span of parasitized voles was reduced by 36% compared to control voles. Parasitized voles had a smaller body size, but a relatively larger heart and spleen than control voles. These results indicate an effect of flea infestation on host life span and our findings strongly suggest that ectoparasites should be taken into account in the studies of host population dynamics.

1. Serial sections were examined of thoraces of a selected strain of Aedes aegypti infected with either Brugia pahangi or subperiodic B. malayi and of wild-caught Mansonia uniformis infected with the latter parasite.

2. A few badly damaged flight muscle fibres are seen in the early stages of infection but many more fibres are destroyed at the time when the mature filarial larvae leave the muscle and enter the haemocoel. In the intervening period, when the larvae are growing within the muscle fibres, only minor damage develops, affecting nuclei and/or mitochondria. The nature of such minor damage and the mechanism of complete muscle breakdown differs in the two species of mosquitoes.

3. It was concluded that the muscle damage was sufficiently severe to explain the significant mortality often observed among experimentally infected mosquitoes at the time when the mature filarial larvae enter the haemocoel.

I am very grateful to Dr W. W. Macdonald, of the Liverpool School of Tropical Medicine, who supplied the Aedes aegypti mosquitoes and who has given invaluable help and advice throughout this study. Mr Cheong Weng Hooi and his colleagues in Kuala Lumpur kindly supplied, and partly processed, the Mansonia uniformis mosquitoes. Miss M. A. Johnson assisted with the care of the A. aegypti colony and Mrs J. Oliver helped with some of the histological work.