A sensitive and specific nitrocellulose (NC) membrane-based dot–ELISA, utilizing a panel of monoclonal antibodies (mAbs), was developed for differentiation between in vitro-derived procyclic forms of Trypanosoma brucei, T. congolense and T. simiae, and epimastigotes of T. vivax. Trypanosomes in suspension were applied onto NC membrane in dots and probed with unlabelled trypanosome species-specific mAbs. Bound mAb was revealed by enzyme labelled anti-mouse IgG and precipitable chromogenic substrate. The assay detected the aforementioned trypanosome species in both single and artificially mixed preparations. Ten T. brucei, 4 T. vivax, 7 T. congolense and 3 T. simiae procyclic stocks and clones from different geographical areas were tested and identified using the specific mAbs in the dot–ELISA which had a specificity of 100%. Some of the T. brucei, T. congolense and Nannomonas-specific mAbs could detect as few as 10 trypanosomes/dot, whilst 1 T. vivax mAb was able to detect a minimum of 100 trypanosomes/dot in monospecies preparations. A concentration of 1 × 104 trypanosomes/μ/dot was eventually determined as ideal for testing in the dot–ELISA. Antigen dots stored at 4 °C under desiccated conditions did not show any loss in activity for up to 90 days. However, when stored under similar conditions at room temperature (17–26 °C), the T. congolense-specific antigen remained unaffected up to 60 days, and then showed decreased activity when tested on day 90.

Pasteuria penetrans is a bacterium with the potential to control plant-parasitic nematode populations; the mechanism of spore adhesion, however, is poorly understood. Attachment assays were performed in media supplemented with various concentrations of fibronectin and in the presence and absence of KSCN which modulates hydrophobic interactions. A reduction in the strength of the hydrophobic effect prevented spores from binding to the cuticle as did 20 μg/ml fibronectin. It was also shown directly utilizing a newly-developed technique which utilizes 3-hexadecanoyl-7-hydro-coumarin as an indicator of the fibronectin binding to the spore surface that the presence of KSCN prohibited binding. This effect was interpreted to indicate that the reduction of binding was the direct result of the influence of hydrophobic interactions between the fibronectin and the spore surface. Western blot analysis of cuticle extracts of Meloidogyne incognita and Caenorhabditis elegans revealed small amounts of fibronectin to be present. Fibronectin, or a similar receptor, present in the cuticle could be responsible for the adhesion of P. penetrans by hydrophobic interactions.

Infection of intermediate hosts with eggs of taeniid parasites results in a larval infestation known as cysticercosis. A number of studies have indicated that cysticercosis is associated with immunosuppression, although little is known about the mechanisms involved. In the present study, mice infected with the larvae of Taenia crassiceps were found to exhibit a pronounced anergy, which preferentially affected T-cells located anatomically close to the parasite. This anergy was linked to late events in the T cell activation pathway; that is, stimulation through the T cell receptor(TCR)/CD complex by Concanavalin-A, or plate-bound monoclonal antibodies (mAb) to TCRαβ or CD3ε, or combinations of phorbol ester and ionomycin (all of which can bypass early membrane-related events), failed to fully activate T lymphocytes. The relative proximity of T cells to the parasite was directly related to upregulation of IL-4 and downregulation of IL-2 production. In addition, the profiles of parasite-specific Abs showed an exclusive increase of serum IgG1 during infection. Taken together, the data suggest that infection of mice with larvae of T. crassiceps alters the balance of CD4+ Th cells by upregulating Th2 and downregulating Th1 cells located in close proximity to the parasite.

A cross-sectional parasitological study of a population of wild olive baboons (Papio cynocephalus anubis), consisting of 5 troops, was conducted in Gombe Stream National Park. Baboons were individually recognizable. Information on age, sex, troop membership, reproductive status, social rank and life-history of each individual baboon could be related to parasite infection. Seven helminth taxa and 2 protozoan taxa were found. All baboons were parasitized by at least 1 taxon. Distributions of helminths were aggregated among hosts. There were significant differences among troops in the prevalence of all but 2 of the recorded helminths. Age had a significant impact on the prevalence and intensity of Strongyloides sp. No significant effect of sex on the prevalence of infection could be detected. There was some indication that female reproductive status was related to Trichuris egg output. In contrast to a previous study, no significant correlations between parasite infection and social rank could be found. Troop membership constituted the predominant factor contributing to heterogeneity of prevalence of infection. This suggests that spatial location and/or genetics may be important in determining levels of parasite infection.

The susceptibility of Brugia malayi and Onchocerca lienalis microfilariae to H2O2 and NO either in cell-free culture or from IFNγ-activated macrophages was examined. In cell-free culture, O. lienalis microfilariae were highly susceptible to H2O2 induced toxicity, exhibiting rapid reductions in motility and viability. The addition of exogenous catalase abrogated H2O2-induced killing. In contrast, B. malayi microfilariae were relatively resistant to H2O2, with concentrations as high as 50 μM having no effect on motility or viability. On exposure to NO, both species showed reductions in motility within 5–30 min, but longer was required to see effects on the viability of microfilariae. Parasites incubated with IFNγ-activated macrophages also exhibited marked reductions in motility and viability. In cultures with B. malayi and activated macrophages, inhibition of these effects was achieved by the addition of either L-NMMA, to abolish NO production, or neutralizing anti-TNFα antibodies. Attempts to inhibit parasite killing by the addition of catalase to macrophage cultures were ineffective. The results of this study show that B. malayi and O. lienalis microfilariae have different susceptibility to H2O2, but are equally affected by exposure to NO. Moreover both species are killed by IFNγ-activated macrophages and in the case of B. malayi, killing is dependent on the generation of NO via TNFα.

A rapid, visually read, dot-ELISA developed for the detection and differentiation of trypanosome species in tsetse flies (Glossina spp.), was field tested alongside the standard fly dissection method on a ranch in south eastern Kenya. Of 104 G. pallidipes dissected, 2 were found to be infected with trypanosomes in their midguts. By the dissection method the infecting trypanosome species could not be identified, as both flies had no salivary gland infections. However, using the dot-ELISA, the 2 flies were shown to be infected with Trypanosoma congolense in their midguts. The midguts of an additional 6 (5·8%) of the 104 G. pallidipes tested positive for T. congolense in the dot-ELISA, even though no trypanosomes were seen on dissection. The infection rate for this fly species as determined using the dot-ELISA, therefore, was 7·7% for T. congolense in midgut infections compared to 1·9% identified by fly dissection. The salivary glands and mouthparts of the 6 additional tsetse flies identified by dot-ELISA were all negative as determined by the 2 techniques. None of 390 G. longipennis flies dissected and examined for trypanosomes in the midgut, salivary glands and mouthparts was shown, by this method, to be infected. Using the dot-ELISA, however, 17 (4·4%) of the flies tested positive for T. congolense in the midgut, whilst the salivary glands and mouthparts of the same flies were negative. Thus, the dot-ELISA appears to be more sensitive than the fly dissection method under field conditions. Moreover, the dot-ELISA can be performed in the field without electricity. It is simple to perform, and was not affected by high ambient temperatures (22–32°C), or by contamination of reactants with dust.

This paper estimates the global burden of lymphatic filariasis based on a review of the published literature on infection and disease surveys. A method for aggregating and projecting prevalence data from individual studies to national, regional and global levels, which also facilitates the estimation of gender and age-specific burdens, is presented. The method weights in favour of the larger, and hence presumbably more reliable, studies and relies on estimated empirical relationships between gender, age, infection and disease in order to correct studies with incomplete data. The results presented here suggest that although the overall prevalence of filariasis cases is 2·0% globally (approximately totalling 119 million cases), the disease continues to be of considerable local importance, particularly in India and Sub-Saharan Africa. Estimates by age and gender clearly show that, unlike other helminth infections, filariasis is mainly a disease of the adult and older age-classes and appears to be more prevalent in males. This work suggests that the derivation of more accurate estimates of the burden of filariasis will require a better understanding of both the epidemiology and the spatial aspects of infection and disease. It also suggests that filariasis is preventable based on a geographically targeted strategy for control.

Endotoxin levels were measured in the blood and cerebrospinal fluid (CSF) of control individuals and 2 groups of patients with African sleeping sickness. Endotoxin levels were markedly elevated in the blood (infected groups mean endotoxin values 40·2 pg/ml and 53·8 pg/ml, compared to control 11·6 pg/ml, P < 0·0001 for both increases) and CSF (infected groups mean endotoxin values 45·8 pg/ml and 50·1 pg/ml compared to control 6·3 pg/ml, P < 0·0001 for both increases) of the patients. The levels were reduced 6 weeks following different drug treatments in the 2 groups (blood levels to mean 33·8·pg/ml and 28·5 pg/ml; CSF levels to 37·4 pg/ml and 27·0 pg/ml). The blood endotoxin values correlated with the CSF values before treatment (r = 0·74 and 0·57 for the 2 groups; P < 0·0001 for both) and after treatment (r = 0·57 and 0·56 for the 2 groups; P < 0·0001 for both). It is concluded that raised endotoxin equilibrates in the blood and CSF compartments, and may contribute significantly to the pathology of sleeping sickness.

There is substantial evidence that host genetic factors play a major role in determining the outcome of infection with many pathogens. Detailed analysis of malaria has identified twelve genes that affect susceptibility in various human populations. However, less attention has been paid to other major infectious diseases where twin studies have identified an important host genetic component to susceptibility. Recent progress in the analysis of the human genome offers exciting prospects for the mapping and identification of new susceptibility and resistance genes for common infectious diseases. Screening of the whole genome in affected sibling pair studies is now feasible by employing highly informative microsatellite markers. In addition, many polymorphic candidate genes have become available for analysis in case-control studies. It is proposed that these new genetic tools offer a powerful approach to the epidemiological analysis of many infectious diseases in humans and supersede traditional genetic approaches to identifying susceptibility genes in mouse models. Progress in characterizing the role of major histocompatibility genes in susceptibility to malaria and other infectious diseases is reviewed before outlining the methodologies for and progress in identifying non-MHC susceptibility genes.

Vitellogenin synthesis by the fat body has been monitored using in vitro culture and immunoprecipitation. This system was found to be efficient for measuring vitellogenin production in both non-infected Tenebrio molitor and those infected with Hymenolepis diminuta. In fat bodies from infected beetles, vitellogenin production was decreased by up to 75% (day 24 post-infection) and, at all times investigated, vitellogenin synthesis was significantly below control levels (days 3–30 post-infection). Incubating fat bodies from control insects with isolated metacestodes indicated that this may be a direct effect by the parasite which is developmental stage-specific. Stage II, but not stage III–IV, nor heat-killed parasites could bring about this decrease in vitellogenin. In addition, these effects may be density dependent within the range of 2–20 parasites per fat body; only 2 metacestodes were necessary to cause a significant decrease. Since metacestodes do not take up vitellogenin, nor limit the amount of [14C] leucine available to the fat body for vitellogenin production, it is conceivable that the parasite produces a potent inhibitor of vitellogenin synthesis, or a molecule which induces cells within the fat body to do so.

The prevalence of various species and subgroups of trypanosomes in the Sinfra area of C⊚te d'Ivoire was determined using the polymerase chain reaction (PCR). Using this technique to amplify specific satellite DNA targets, it was possible to identify developmental-stage trypanosomes in the midguts and the proboscides of tsetse without expansion of parasite populations. The predominant tsetse species in the area was Glossina palpalis, while G. pallicera and G. nigrofusca were also present. Microscopical examination of 811 non-teneral flies revealed an infection rate of 14% in midguts and/or proboscides. Three subgroups of Trypanosoma congolense (Savannah, Forest & Kilifi), T. simiae, T.godfreyi, West African T. vivax and T. brucei ssp. were identified using PCR. T. congolense Forest was the most abundant of the Nannomonas trypanosomes. Approximately 40% of all infections were mixed, and there was a significantly higher prevalence of apparently mature T. brucei ssp. trypanosomes than has previously been reported. The present study demonstrates that PCR facilitates the easy identification of mature trypanosome infections in tsetse, providing a reliable estimation of trypanosomiasis challenge.

Models have predicted that directly transmitted macroparasites may influence the abundance of forage plants in herbivore grazing systems by reducing the food intake of their host. Evidence of parasite-induced alterations in host food intake is, however, limited mainly to sheep, cattle and laboratory rodents. We estimated the effect of naturally acquired parasite infections on the appetite of reindeer. Food intake was significantly lower in infected reindeer compared to animals in which the parasites had been experimentally removed. Among the infected animals there was a significant negative relationship between intensity of the directly transmitted macroparasites (i. e. gastrointestinal nematodes) and mean food intake, indicating that the lower food intake was caused by these parasites. The time-specific onset of depression in food intake is also consistent with seasonality in the pathogenic effect from gastrointestinal nematodes. This shows that parasite-induced changes in herbivore food intake is not restricted to agricultural systems, and implies that parasites may have impact on the dynamics of a wide range of herbivore plant communities.

We investigated the timing of seasonal activity of the seabird tick Ixodes uriae by examining engorgement rates of off-host ticks and the prevalence of infested common guillemots (Uria aalge) and kittiwakes (Rissa tridactyla) on the Isle of May, Scotland. More than 99% of I. uriae fed during the period late April to early August which coincided with the seabird breeding season. No ticks fed in the autumn and winter, and none over-wintered in the engorged state. Peak tick feeding activity was during the hosts' incubation period in May and early June, when over 70% of birds were parasitized. Few ticks were active when guillemot chicks were being brooded, or during the early part of kittiwake chick-rearing. Adult female tick feeding peaked in the first week of June, while nymphal and larval activity peaked in the third week of May. Adult female ticks fed later than nymphs on guillemots and larvae on both guillemots and kittiwakes; however, there was considerable temporal overlap in the feeding periods of the 3 stages on both hosts. All larvae and nymphs fed in May and early June, but up to 18% of adult females remained unengorged at this time, and probably deferred feeding by 1 year. A small proportion of individuals which fed as larvae in May appeared to feed again as nymphs in July of the same season, thereby shortening their life-cycle by 1 year. The duration of the life-cycle of I. uriae on the Isle of May is typically 3 years but varies from 2 to 4 years. Ticks feeding on guillemots did so significantly earlier than those feeding on kittiwakes, possibly because guillemots start breeding earlier in the year. Breeding of both guillemots and kittiwakes was later in 1994 than in 1993. This delayed the activity of all tick stages on kittiwakes and adult female ticks on guillemots, but did not affect the timing of activity of nymphs or larvae feeding on guillemots.

Mice inoculated with microfilariae of the filarial nematode Onchocerca lienalis clear their parasites over a period of 3–4 months and are highly resistant to re-infection. We have investigated the comparative roles of the eosinophil, macrophage and neutrophil in effecting this parasite clearance, employing agents specifically to perturb cell function in vivo. Using the anti-IL-5 monoclonal antibody TRFK-5, we show that eosinophils are of primary importance in effecting resistance to re-infection. Ablation of macrophages (with carbon) and neutrophils (with the monoclonal antibody NIMP-R14) had no effect on parasite clearance following re-infection. Neutralization of these 3 cell types during a primary infection showed that while the removal of both eosinophils and macrophages caused a small but significant delay in parasite clearance, the depletion of neutrophils had no effect. This report describes the first direct evidence for eosinophil-mediated killing of microfilariae in vivo, and is consistent with Th-2 cell responses previously described in this model.

A condition with a weak genetic component, but a strong correlation with age, is the summarizing of conferences. It is an art form often resembling the expression of gratitude by a bride on the receipt of her sixth electric kettle. However, our gratefulness now is genuine, because we have been given a wonderfully interesting series of papers. Their authors were chosen with skill to show the manifold variety of genetic interactions between parasites and hosts. The range of these interactions would daunt even the strongest-hearted summarizer, and my task can only be managed by a ruthless subjectivity.

Roach (n = 81) caught on 23 May, 1992 from oligotrophic, unpolluted Lake Peurunka were kept in cages over a 2-week period before moving half of the fish to nearby Lake Vatia, which is influenced by pulp mill effluents. Before moving the fish gill parasites were examined from 9 fish; 5 Dactylogyrus species, Gyrodactylus sp. and Paradiplozoon homoion were found, the main components of the infracommunities being dactylogyrids. Afterwards, 5 fish from each lake were studied weekly between 1 July and 17 August. Metazoan parasites were recorded from gill arches divided into 4 sections along the dorso-ventral axis. D. crucifer, D. nanus, D. micracanthus and D. suecicus occurred in both lakes throughout the study. Gyrodactylus sp., Ergasilus briani and P. homoion occurred in low numbers in both lakes. Differences between the lakes were seen in the prevalence of D. micracanthus which was higher in Lake Vatia and significantly increased abundances of D. crucifer, D. nanus and D. micracanthus in Lake Vatia. Abundances of other Dactylogyrus species remained, on average, at the same level in both lakes. No change during the experiment was recorded in the location of the two most common species, D. crucifer and D. nanus, on the host gills in either of the lakes, both species favouring the 2nd and 3rd gill arches and especially the inner parts of the gills. The apparent lack of competition between the 2 most common Dactylogyrus species indicates that no limitation of resources on roach gills occurred. This was also confirmed by the increased overlapping indices with increasing abundances between the two most common species in Lake Vatia.

An in vitro assay was used to determine the effects of Sanguinicola inermis adults and cercariae on the polarization responses of pronephric leucocytes of carp. Leucocytes were isolated and exposed to live adult flukes or cercariae for up to 48 h. Differences in polarization responses were related to the presence of the parasite, the presence or absence of carp serum and the time of incubation. The mean proportions of cells exhibiting polarization in unstimulated controls ranged from 5 to 30% over the experimental period. Within 15 min of exposure to adults or cercariae, significant increases in mean polarization responses were observed of up to 75% and levels remained higher than control values for over 24 h. Overall, the presence of normal carp serum, either untreated or heat inactivated, did not enhance the polarization responses of leucocytes incubated with only cercariae or adults. However, between 0·25 and 3 h, the presence of carp serum with cercariae significantly enhanced polarization responses when compared with cells incubated with cercariae alone.

The presence of retinol-binding protein (RBP) activity in Onchocerca cervicalis adult worms and interaction with ivermectin has been studied using high pressure size exclusion chromatography (HPSEC). Four distinct peaks of [3H]-retinol incorporation were obtained corresponding to approximate molecular weights of 150, 67, 19·7 and 4–6 kDa, the 2 smaller Mr peaks accounting for most of the binding activity. Competition for binding using non-labelled retinol at 200-fold molar excess indicated that specific binding of retinol occurred only to the 19–7 kDa fraction. Competition by ivermectin also inhibited binding of [3H]-retinol to the third peak. Following incubation with [3H]-ivermectin & peaks of similar molecular weights were also detected by HPSEC in soluble adult worm homogenate, However, in this case the 150 kDa fraction was most prominent. Both non-labelled ivermectin and non-labelled retinol at 200-fold molar excess reduced binding of [3H]-ivermectin to all & fractions. These data indicate that the putative Onchocerca RBP has an approximate molecular weight of 19·7 kDa, that retinol also binds to 3 additional fractions non-specifically, that the pattern of binding of ivermectin to adult worm material is quantitatively and qualitatively different from the binding exhibited by retinol, and that ivermectin interferes with the binding of retinol to the 19·7 kDa Onchocerca protein.

Dirofilaria immitis is an important filarial parasite of dogs and cats, and a useful model for human filariasis. Current diagnostic tests for heartworm infection in animals rely on the presence of fecund female worms (usually found 6·5 months post-infection or later) and therefore fail to detect pre-patent infections. Putative pepsin inhibitors from 2 filarial parasites of humans namely Onchocerca volvulus (Ov33, Oc3.6, OvDSB) and Brugia malayi (Bm33), have been shown to be useful in diagnosis of onchocerciasis and lymphatic filariasis, respectively. Previous studies have suggested that a homologue exists in D. immitis (DiT33), which may have potential in diagnosis of heartworm infection. In this study, the isolation and characterization of a cDNA clone encoding DiT33 is described.‡ This cDNA contains 12 bases of the nematode-specific 22 nucleotide spliced leader sequence and encodes a 26·4 kDa-protein with a high level of similarity (87–89%) to other filarial members of the family. DJT33 was over-expressed in E. coli as a fusion with the maltose-binding protein and serological analysis was performed using a panel of clinically defined dog sera. The findings of this study indicate that DiT33 is a promising antigen for the early detection of D. immitis and may be a valuable tool in the control and management of heartworm infection.

Parasites are generally found aggregated among hosts. In this paper we propose a method for measuring aggregation at different scales in the host population. We use the method to characterize the pattern of aggregation of the tick Ixodes uriae on chicks of its seabird host, the Kittiwake Rissa tridactyla. We found evidence of aggregation at the among-nest scale, but not among chicks within nests. This shows that the processes leading to aggregation occur at a higher scale than the nest. The methods we develop provide a way to compare parasite aggregation at different scales in a quantitative fashion and can be applied in a large number of epidemiological studies.