Asymptomatic malaria parasitaemia is prevalent among schoolchildren in many countries. The relationship between asymptomatic parasitaemia and children's cognitive functions was examined in a case control study and then in a natural experiment. A group (n = 445) of asymptomatic parasitaemic boys were compared with a group of non-parasitaemic boys (n = 142) matched for grade and school on their performance on a battery of cognitive tests. Two weeks later the parasitaemic children were re-screened and 150 children of those who remained parasitaemic were matched for grade and school with 150 children who were no longer parasitaemic. These children were then re-tested and their cognitive function compared. Initially, after controlling for age, socio-economic background and nutritional status the parasitaemic children performed worse than the non-parasitaemic children in fine motor function tests. There was no difference in change in cognitive test scores between those who became non-parasitaemic and those who remained parasitaemic. However, children who initially had the highest parasite density improved the most in 2 fine motor tests and a picture memory test. We were unable to show a benefit from losing parasitaemia over a 2 week period, but it remains possible that parasitaemia may affect cognition and longer term trials should be conducted.

The parasite, Neospora caninum is an important cause of abortion in cattle. It is transmitted vertically or horizontally and infection may result in abortion or the birth of a live, healthy but infected calf at full-term. Only a proportion of infected cattle abort and the pathogenesis of abortion is not understood. Groups of cattle were infected with 107N. caninum tachyzoites intravenously at different times relative to gestation. Intravenous inoculation was chosen to reproduce the putative haematogenous spread of N. caninum following either recrudescence of endogenous infection or de novo infection. In all cattle, infection was accompanied by high γ-interferon and lymphoproliferative responses, and a biased IgG2 response indicating that N. caninum infection is accompanied by a profound Th1 helper T cell-like response. Infection at 10 weeks gestation resulted in foetopathy and resorption of foetal tissues 3 weeks after infection in 5 out of 6 cows. Infection at 30 weeks gestation resulted in the birth of asymptomatic, congenitally-infected calves at full term in all 6 cows, whereas the 6 cows infected before artificial insemination gave birth to live, uninfected calves. These results suggest that the reason some cows abort is related to the time during gestation when they become infected or an existing infection recrudesces.

We have studied the activity of the polyether ionophorous antibiotic monensin against the cyst form (bradyzoite) of Toxoplasma gondii. The experiments were conducted in vitro using 2 methods; cysts produced either in mice or in cell culture were exposed to monensin in vitro, and the infectivity of the parasites was then assessed in vivo or in vitro. The data obtained from these 2 systems of evaluation showed that monensin inhibits the infectivity and the viability of the bradyzoites. Its activity was time and concentration dependent. The first effects were observed at very low drug concentrations (i.e. 0·0001 μg/ml). Immunofluorescence and electron microscopy analysis showed significant cytological alterations of the monensin-treated bradyzoites: they were swollen, had a large number of vacuoles in their cytoplasm and were found lysed at higher concentrations in ionophore.

Promastigotes of Leishmania mexicana mutants lacking the multicopy CPB cysteine proteinase genes (ΔCPB) are markedly less able than wild-type parasites to infect macrophages in vitro. ΔCPB promastigotes invade macrophages in large numbers but are unable to survive in the majority of the cells. In contrast, ΔCPB amastigotes invade and survive within macrophages in vitro. This extreme in vitro stage-specific difference was not mimicked in vivo; both promastigotes and amastigotes of ΔCPB produced lesions in BALB/c mice, but in each case the lesions grew considerably more slowly than those caused by wild-type parasites and only small lesions resulted. Inhibition of CPB in situ using cell-permeant peptidyldiazomethylketones had no measurable effect on parasite growth or differentiation axenically in vitro. In contrast, N-benzoyloxycarbonyl-phe-ala-diazomethylketone reduced the infectivity of wild-type parasites to macrophages by 80%. Time-course experiments demonstrated that application of the inhibitor caused effects not seen with ΔCPB, suggesting that CPB may not be the prime target of this inhibitor. The data show that the CPB genes of L. mexicana encode enzymes that have important roles in intracellular survival of the parasite and more generally in its interaction with its mammalian host.

The sex ratio of adult worms has been observed biased towards males in Schistosoma mansoni under natural conditions. The origin of this bias is unknown. This paper determines whether males are more infective than females under controlled experimental bisexual conditions, and hence if the sex ratio is male-biased as a consequence of this. The male and female cercarial infectivities in uni- and bisexual vertebrate host infections using a range of controlled cercarial sex ratios were studied. The results showed that, in experimental unisexual infections, male cercariae were more infective than females, and that in experimental bisexual infections, male cercarial infectivity was similar to that of female, irrespective of cercarial sex ratio. Furthermore, cumulative male and female cercarial infectivity was maximal when sex ratio was equilibrated. The unbiased sex ratios obtained in our experimental bisexual infections are discussed in terms of behavioural and/or biochemical male–female interaction. Alternative explanations of the natural biased sex ratio are proposed.

A schistosome infection is initiated when the parasite penetrates the skin of a susceptible host. Relatively large quantities of protein are released by transforming cercariae compared to later larval stages. This represents the first parasite material to which the host's immune system is exposed, yet little is known about the proteins which are released during the first few hours post-transformation. We have shown that antiserum raised against such molecules was capable of imparting protection against a schistosome challenge infection upon passive transfer to naïve mice. By screening a cercarial cDNA library with this serum, 38 positive clones were identified. Sequence analysis showed these to represent 8 different molecules which included Schistosoma mansoni 21·7 kDa antigen, calcium-binding-protein and the vaccine candidate glutathione S-transferase (Sm28GST). In addition, 5 clones were isolated, 1 of which had significant homology to many cytochrome C proteins, another with leukocyte elastase inhibitors and 3 which represented novel molecules. Four clones were expressed in a prokaryotic high-level expression vector, sera produced against each purified recombinant protein and used subsequently to probe Western blots and parasite sections. The leukocyte elastase inhibitor homologue and 2 unknowns induced significant proliferation by lymph node cells recovered from mice vaccinated with irradiated cercariae. More strikingly, the 2 novel proteins stimulated very high levels of interferon γ (IFNγ) secretion both by lymph node cells and those recovered by broncho-alveolar lavage from the lungs of vaccinated mice. Such results will be discussed in the context of vaccine development.

Parasites should be better at infecting hosts from sympatric populations than allopatric populations most of the time (parasite local adaptation). In a previous study of a population of snail parasites (Microphallus sp.) from Lake Alexandrina, New Zealand, we found that Microphallus was more infective to snails (Potamopyrgus antipodarum) in shallow water but not in deep water. Here, we repeated the original study and also monitored the development of the parasite. We found that parasites from shallow water were more infective to hosts from shallow water and developed more rapidly in these hosts. In contrast, parasites from deep water were not more infective to hosts from deep water and did not develop more rapidly in them. These results suggest clinal variation in the susceptibility of these snails, with shallow-water snails more susceptible than deep-water snails. We offer 2 possible explanations for these results. First, gene flow in the Microphallus population is primarily from shallow to deep water, leading to an asymmetric pattern of local adaptation. Alternatively, snails from shallow water may be more susceptible for reasons independent of gene flow, perhaps due to differences in host condition between habitats.

Isozyme analysis (12 enzymes: 14 loci) was conducted on 99 isolates of Trypanosoma cruzi: 77 from Guatemala, 5 from Mexico and 17 from South American countries. Analyses of 4 population-genetic indices were undertaken to assess the possibility of genetic exchange occurring among Guatemalan isolates. The results provide evidence for a degree of genetic exchange occurring among isolates from this relatively small geographical area. Previous studies of population genetics on T. cruzi might have failed to detect this phenomenon because they tended to use isolates originating far from one another, rendering gene exchange unlikely for geographical reasons. Phylogenetic data, presented here, show considerable differences in genetic structure between Central and South American isolates, suggesting that different biological and clinical properties might be expected. For example, there are differences in clinical syndromes between Central and South America, a situation discussed further here.

The present study describes a novel heat-stable, water-stress-related protein with a molecular mass of 47 kDa (designated Desc47) in the entomopathogenic nematode Steinernema feltiae (IS-6). The protein was accumulated about 10-fold (from 7·84±1·85 to 74·09±4·35% relative content level [RCL]) in dehydrated clumps of infective juveniles (IJs), which had lost 34·4% of their initial water content (from 65·1±1·7% to 42·7±0·72%) in a desiccation-tolerance-inducing treatment (97% relative humidity [RH] for 3 days). The appearance of Desc47 was accompanied by trehalose accumulation (from 300 to 600 mg trehalose/g protein) during the process of inducing the IJs into a quiescent anhydrobiotic state. A second cycle of IJ dehydration did not alter the RCL of Desc47 (79·3% for the first cycle and 73·3% for the second cycle). Desc47 retained its high RCL (69·7%) in rehydrated active IJs for 3 days, reaching 51·2% of its initial RCL only after a week. No homology to other known proteins was found by mass-spectrometry electrospray-ion-trap analysis. However, of the 5 sequences obtained from the protein (ranging from 11 to 21 amino acids), the 21-amino-acid peptide NVASDAVETVGNAAGQAG(D/T)AV showed excellent homology (74% identity in 19 amino acids) to the cold-responsive protein COR14b (g6564861) from Triticum aestivum. In the Caenorhabditis elegans predicted proteome database search, the N21 yielded the first-best identity score (59% identity in 17 amino acids) to the CE-LEA homologue protein (g2353333). In plants, COR and LEA are related proteins, heat-stable, which are expressed in response to both dehydration and cold acclimation. The implication of the involvement of Desc47 and the osmoprotectant trehalose in the desiccation-tolerance mechanisms of S. feltiae is discussed.

The frequency distribution of first generation, Steinernema feltiae Filipjev parasitic stages was over-dispersed with the majority of hosts containing few or no parasitic stages, whilst a few hosts contained a great many. Because of high extraction efficiency, the frequency distributions of the parasitic stages and the infective stages in the soil were assumed to be directly related. To explain the frequency distribution of the parasites it was therefore necessary to account for the frequency distribution of the S. feltiae infective stages in the soil. The infective stages were spatially aggregated into 30 cm diameter patches at the site of host death. These patches were randomly distributed approximately 1 m apart. At the 1 m scale, the pooled counts of infective stages were randomly distributed. Thus, in contrast to the frequency distributions, the spatial structuring of S. feltiae changed with the spatial scale of the interaction. This dynamic spatial structuring means that the majority of samples taken would contain few or no infective stages, whilst a few soil samples would contain a great many. Thus, the spatial structuring of the infective stages generates the over-dispersed frequency distribution of the S. feltiae in the soil. Hosts, encountering infective stages from this spatial distribution will, therefore, show an over-dispersed frequency distribution of S. feltiae parasitic stages.

It recently became clear that adult eo- and palaeacanthocephalans parasitizing fish can bioconcentrate several heavy metals to significantly higher concentrations than the tissues of their definitive hosts. Following this discovery the lead accumulation of the archiacanthocephalan Moniliformis moniliformis was investigated using experimentally infected male Wistar rats of the CD-M-strain. The worms were allowed to grow up for 4 weeks post-infection followed by a 3 weeks oral lead exposure of the rats. After the exposure period the rats were killed and the metal levels were determined in muscle, liver, intestine and kidney of the rats as well as in different organs of female and male acanthocephalans. Lead concentrations were found to be highest in female M. moniliformis followed by the kidneys of the rats. Male worms contained approximately the same lead concentration as the hosts' kidneys. Lead analysis of the worms' organs revealed the highest lead concentration in the eggs of female acanthocephalans, followed by the cement gland of male Worms. Whilst the lead burden of the presoma was higher than that detected in the kidneys of the rats, the lead content of the metasoma was even lower than in the kidneys. A lead uptake of M. moniliformis from the intestinal lumen of the host became apparent as the faeces of infected rats contained significantly less lead compared to the uninfected conspecifics. Thus, this study reveals that lead accumulation also occurs in archiacanthocephalans parasitizing mammals. But the degree of metal bioconcentration is considerably lower compared to eo- and palaeacanthocephalans in fish. Anyway, due to a lack of adequate sentinel species in terrestrial biotopes the host–parasite system rat– M. moniliformis appears to be a useful and promising bioindication system especially in urban ecosystems in temperate regions.

The relationship between faecal nematode egg count and the number of adult Telordasagia circumcincta was examined in 508 naturally infected lambs at 6–7 months of age. The relationship was found to be convex and was empirically described by a gamma-type function of the form y = anb e−cn, where n is the number of adult nematodes present and y is the number of nematode eggs per gramme (epg) of faeces. This equation predicted that the peak expected egg count (277 epg) would occur at a worm burden of 2167 adult worms. The absence of a linear relationship between egg counts and worm numbers is consistent with the existence of severe density-dependent constraints on the fecundity of T. circumcincta.

The nervous system of young and adult Amphilina foliacea was studied with immunocytochemical, electron microscopical and spectrofluorometrical methods. The general neuroanatomy is described in detail. New data on the structure and development of the brain were obtained. The 5-HT and GYIRFamide-immunoreactivities occur in separate sets of neurones. The innervation of the reproductive organs is described. The fine structure of 2 types of neurones in the CNS, a sensory neurone, a ‘glial’ cell type, the neuropile and the synapses are described. The level of 5-HT varies between 0·074 and 0·461 μg/g wet weight. This is the first detailed study of the nervous system of A. foliacea. Earlier data on the structure of the nervous system in A. foliacea published in Russian are introduced into the discussion. The study provides data that can be used when considering the phylogenetic position of Amphilinidea.

Nuclear-encoded large-subunit ribosomal DNA sequences were used to infer a phylogenetic hypothesis for 17 taxa (16 nominal species) of the genera Contracaecum and Phocascaris. Phylogenetic trees based on these data have been used to assess the validity of the taxonomic distinction between these genera, which was based on the presence or absence of certain structural features, rather than on explicit hypotheses of evolutionary history. Phylogenetic hypotheses based on parsimony, likelihood, and neighbor-joining analyses of these sequence data strongly support the hypothesis that species of Phocascaris are nested within the clade of Contracaecum species hosted by phocid seals, and are more closely related to species of the Contracaecum osculatum complex than to other Contracaecum species. Alternative tree topologies representing Phocascaris as not nested within the C. osculatum complex were significantly worse interpretations of these sequence data. Phylogenetic analysis also provides strong support for the monophyly of all taxa (Contracaecum and Phocascaris) from phocid seals, which is consistent with Berland's (1964) proposal that such species form a natural group; however, his proposal to recognize all species in phocid seals as Phocascaris, with all species from birds as Contracaecum would result in a paraphyletic Contracaecum, according to the molecular phylogenetic hypothesis.