The in vitro transformation of the miracidium to the mother sporocyst of Schistosoma margrebowiei was initiated by placing the miracidium in mammalian physiological saline. The transformation occurs in stages: the cilia cease beating; the ciliated plates become detached from the intercellular ridges and underlying muscle layers; the intercellular ridges spread over the body surface eventually forming a new tegument; the sporocyst changes from an ovoid to a tubular shape in about 48 h at room temperature. The surfaces of the miracidium, sporocyst and cercaria of S. margrebowiei display stage-specific carbohydrates on their surfaces as indicated by lectin staining. Ricin120 stains the cilia alone of the miracidium whereas peanut agglutinin stains the larval surface except for the cilia. The intercellular ridges of the miracidium stain with concanavalin A and wheat germ agglutinin, and these lectins stain the entire surface of the mature mother sporocyst. The cercaria is the only larval stage which stains positively with asparagus pea lectin. Bulinus nasutus is incompatible with Schistosoma margrebowiei; the haemolymph of this snail contains an agglutinin which agglutinates a wide variety of mammalian erythrocytes including those of human ABO blood groups. The haemagglutinin titre of B. nasutus plasma is reduced after incubation with miracidia of S. margrebowiei indicating that the agglutinin is absorbed onto the surface of this larval stage but not that of the mother sporocyst or cercaria. The possible roles of agglutinins in host–parasite interactions together with the significance of the differences in the surface carbohydrates of the larval stages are discussed.

Onchocerca ochengi is a common parasite of cattle in savanna areas of West Africa and its adult stages inhabit intradermal nodules. We have exploited the accessibility of the macrofilariae to examine quantitative and qualitative differences in both the macro- and microfilariae (mff) in relation to age in naturally infected cattle. Autochthonous cattle at a site on the Adamawa plateau, North Cameroon, situated near a perennial Simulium damnosum s.l. breeding site, were examined in three age groups (1·5–2·5, n = 34; 3–5, n = 39 and ≥ 8 years old, n = 21). Skin mff densities were assessed from ventral skin biopsies, total body counts of nodules were done by palpation and 1–4 nodules were excised from positive animals, collagenase digested and the numbers and condition of macrofilariae determined. Embryogenesis profiles (embryograms) were done on a representative number of female worms in each age group. The overall prevalence of infection in the study population was 71% for mff and 85% for nodules. Nodule prevalence increased significantly from the 1·5–2·5 to 3–5 years old groups, but no other inter-age prevalences were significantly different. While the mean number of nodules per animal increased with age, geometric means (gm) 3·7, 15·5 and 23·1, the skin mff density maximized in the middle age group and decreased significantly in the old animals (gm 15·8, 44·7 and 11·4 mff/g, difference between latter two parameters significant, P < 0·05). Moreover, there was a significant correlation (P < 0·01) between individual nodule counts and skin mff densities in the mid-aged group but not in the oldest group. Examination of nodules showed that nodule counts equated to numbers of female worms, since almost all of 133 nodules examined contained only 1 live female worm. Although the proportion of old female (and male) worms increased with the age groups – 2 (0), 30 (28) and 54 (32)% – the proportion of gravid females (52·8, 48·8 and 58·0%) and the mean number of viable embryos in these females (82·5 × 103, 76·0 × 103 and 102·1 × 103) did not change with age. Thus, due to the higher nodule counts, a calculated productivity index reflecting the potential mff production per animal was 8 times higher in the ≥ 8-year-old animals than in the youngest and 3 times higher than in the middle group, which sharply contrasts with the decline in skin mff density. Although further research is required, these findings may indicate acquired immunity directed against microfilariae. They demonstrate the potential of O. ochengi infections in cattle to investigate a number of aspects of the host–parasite relationship which may have relevance to human onchocerciasis.

Molecular characterization of the Plasmodium falciparum genome has led to identification of polymorphic loci and the mechanisms generating genetic diversity in this parasite. This information has resulted in the development of molecular methods to type parasite diversity in the field. Consequently, we are now in a position to describe the population genetics and dynamics of P. falciparum. The limited number of field studies that have been conducted to date have revealed an extraordinary degree of genetic diversity in natural parasite populations. Heterozygous recombination which occurs during meiosis appears to be one mechanism for generating genetic diversity. The rate at which such recombination occurs in natural parasite populations defines the genetic structure of the parasite population and can influence the ability of the parasite to respond to selection pressure. The high frequency of single genotype infections and the female-biased gametocyte sex ratios found in hyperendemic malaria areas suggest that self-fertilization occurs frequently. Population- wide surveys of allele frequencies in endemic areas have, however, shown no evidence of linkage disequilibrium and are consistent with a panmictic population structure. We argue that these studies have only sampled symptomatic infections, within which rare or recombinant genotypes may be disproportionately represented. They also take no account of the spatial structure of P. falciparum populations. Systematic investigations of the amount of heterozygosity in small areas as part of population-wide surveys are required to define the genetic structure of P. falciparum populations. Population dynamic studies which consider genetic heterogeneity of P. falciparum have shown fluctuations of different serotypes in space and time. The host immune response appears to play an important role in generating these dynamics. Integrated field and laboratory studies, which consider the interaction between population genetics and dynamics, will be necessary to describe the population biology of P. falciparum.

The cDNA probe, Lmet2, was labelled with digoxigenin and used in a chemiluminescent system to detect fewer than 100 membrane-immobilized Leishmania parasites. The probe was found to hybridize primarily with members of the L. donovani complex but a slight cross-reaction was also observed with greater than 5 x 104L. major. This cross-reaction was reduced by hybridizations in 50% formamide at 37 °C. Formamide also significantly reduced non-specific binding of the digoxigenin-labelled probe to the membrane support which, in hybridizations without formamide, masked the specific hybridization signal. This background was not observed with the corresponding radio-isotope labelled probe. With hybridizations in formamide the sensitivity achieved by the chemiluminescent system after exposure to film for 3 h was greater than that achieved by the isotopic system even after autoradiography for 24 h.

The eggs and oncomiracidia of two species of the capsalid genus Encotyllabe are described. These parasites were identified tentatively as E. caballeroi Velasquez, 1977 and E. caranxi Lebedev, 1967. If these identifications are correct, then E. caballeroi is recorded from two new hosts, Gymnocranius audleyi and the nemipterid Scolopsis monogramma, and E. caranxi from a new host, Pseudocaranx dentex, and a new locality, Heron Island, Queensland, Australia. Encotyllabines have not previously been recorded from fishes of the family Nemipteridae. The eggs of the two parasites failed to hatch spontaneously and did not hatch when exposed to a variety of potential hatching stimuli, but the oncomiracidia within survived for many weeks. Oncomiracidia expelled from eggs by cover-slip pressure are unciliated and possess a saucer-shaped haptor like that of other capsalids with three pairs of median sclerites and 14 marginal hooklets. The paths of tendons associated with the accessory sclerites and the presence of haptoral loculi suggest that encotyllabines are related to the trochopodines. Observations on a single juvenile specimen of E. caballeroi show that the accessory sclerites and the tendons are lost early in development and that one pair of hamuli (probably the posterior pair) ceases to grow early in post-oncomiracidial life. The loculi persist a little longer but also disappear before full sexual maturity is reached.

In situations where malaria eradication is not an option in the foreseeable future the emphasis must be on the control of morbidity and mortality due to malaria. Under such circumstances drawing a distinction between malarial parasitization and malarial disease may be important for workers in both field and laboratory. This concept is explored from the points of view of the epidemiological picture of malaria in endemic populations, the factors which may influence progression to disease and the processes which mediate disease.

The localization of adult Schistosoma mansoni originating from Guadeloupe (West Indies) was analysed in the natural host Rattus rattus in experimental and natural infections. A transfer of schistosomes from the porto-mesenteric system to the lungs occurs between the 4th and the 20th weeks post-infection, with a peak between 4 and 8 weeks; it should be noted that the worms start laying eggs at 4 weeks post-infection. In both experimentally and naturally infected R. rattus the relocation of schistosomes in the lungs is correlated with the total worm burden. Between 6 and 12 weeks post-infection some adult worms and numerous eggs are trapped in the liver. A high mortality in the worm population coincides with the migration of the parasites from the porto-mesenteric veins to the lungs. In re-infected R. rattus a relationship appears between the presence of schistosomes in the lungs and resistance to reinfection. It is concluded that, as already postulated previously in mice, resistance to re-infection in the natural host R. rattus is at least partly correlated with the hepatic pathology caused by the schistosomes of the primary infection.

Naturally occurring immunoglobulin M antibodies to Toxoplasma gondii in human sera were examined for their ability to enhance toxoplasmacidal activities of human neutrophils to show a role of natural antibodies in immunity to the early stage of Toxoplasma infection in humans. Neutrophils were mixed with tachyzoites at a ratio of 1:5 in the presence or absence of antibodies and were cultured for 1–18 h for microscopical examination. The count of tachyzoites phagocytosed in 200 neutrophils within 1 h was significantly higher in the presence of sera with natural IgM antibody levels of > 0.8 than those of < 0.1, with 2.4- to 2.9-fold differences (P < 0.02). The total tachyzoite counts at 18 h decreased to 26–39% of those at 1 h in the presence of natural IgM antibody levels of > 0.8 (P < 0.01), while the counts at 1 h consistently increased by 18 h in the absence of natural IgM antibodies. These results indicate that relatively high levels of natural IgM antibodies enhanced phagocytic and microbicidal activities of neutrophils against Toxoplasma. The enhancement was dose dependent, and was also weaker than that obtained with the same dilution of sera from individuals with chronic or acute infections. Live and dead tachyzoite counts in infected neutrophils suggested a quicker effect of natural IgM antibodies than IgG antibodies elicited by infection.

The mechanism whereby praziquantel produces a contraction and subsequent flaccid paralysis (a loss of sensitivity to subsequent stimuli) of Schistosoma mansoni in a medium containing an elevated Mg2+:Ca2+ ratio was investigated. In RPMI, praziquantel produced a concentration-dependent tonic contraction of the parasite with an EC50 of 200 nM. Magnesium inhibited the contraction in such a manner as to convert the tonic contraction to a phasic one without altering the peak force generated. The Mg2+-dependent block was non-competitive with praziquantel but was competitive with extracellular Ca2+, ratios of 7·5:1; Mg2+: Ca2+ being needed to inhibit the tonic contraction and to induce flaccid paralysis. Flaccid paralysis was associated with a reduced ability of the parasite to take up 45Ca2+ from the bath compared to parasites that had not entered into flaccid paralysis and flaccid paralysis was reversible. Recovery from flaccid paralysis was accelerated by treatments that are expected to increase Ca2+ uptake by the parasite. At a concentration of 500 nM, praziquantel produced 2 distinct phasic contractions in intact parasites incubated in an elevated [Mg2+] medium but only 1 phasic contraction in parasites lacking their surface tegumental membranes. In zero Ca2+ I-RPMI, 10 μM praziquantel produced a phasic contraction of intact parasites but did not stimulate contraction of detegumented parasites until Ca2+ was reintroduced into the bath. These results indicate that praziquantel interacts with specific Ca2+-permeable sites in the tegumental and sarcoplasmic membranes of the parasite and that under these conditions of elevated Mg2+:Ca2+ ratios, these sites become blocked by Mg2+, leading to flaccid paralysis of the parasite.

Patients infected with Schistosoma mansoni showed an abnormal response to a test dose of tryptophan, with little increase in the urinary excretion of kynurenine, hydroxykynurenine, xanthurenic and kynurenic acids, N1-methyl nicotinamide, methyl pyridone carboxamide, 5-hydroxytryptamine or 5-hydroxyindoleacetic acid. In contrast to previous reports, this is different from the pattern of tryptophan metabolism seen in vitamin B6 deficiency. Furthermore, the patients' plasma concentrations of pyridoxal phosphate were within the reference range, and supplementation for 5 days with 20 mg vitamin B6/day did not affect tryptophan metabolism. Treatment with a single dose of Praziquantel resulted in a substantial restoration of normal tryptophan metabolism. In mice infected with S. mansoni there was a similar impairment of tryptophan metabolism, as shown by considerably reduced formation of 14CO2 after the administration of a tracer dose of [14C]tryptophan. Again, the administration of vitamin B6 supplements did not correct tryptophan metabolism in the mice. Treatment with Praziquantel resulted in substantial restoration of the production of 14CO2 from [14C]tryptophan. There was no evidence of vitamin B6 deficiency (as determined by erythrocyte aspartate aminotransferase activation coefficient) associated with infection in the mice, although there was a redistribution of pyridoxal phosphate between tissues, with a reduction in the concentration of liver, spleen and kidney, and an increase in skeletal muscle.

Dactylogyrus (Monogenea) species were studied on the gills of roach (Rutilus rutilus) from a total of 293 fish in 3 interconnected lakes in Central Finland. One of the lakes is eutrophic and polluted by a paper and pulp mill, one is eutrophic and one is oligotrophic. Nine Dactylogyrus species were found. The structure of the monogenean communities was analysed at the component and infracommunity level and the Dactylogyrus fauna of the lakes was compared. Although it was found that the component communities tended to be very similar there were also differences. The abundance and diversity were highest in the polluted lake; the infracommunities of this lake were considerably more often dominated by Dactylogyrus micracanthus and the similarities between the infracommunities were higher within the lakes than between the lakes. It is suggested that these differences are due to the differing water quality of the lakes. The nature of the monogenean community, its possible interactivity, is also discussed.

The basic tenet of the immunological perspective of fuiarial disease is that differential immune responsiveness among individuals exposed to infection results in the different clinical manifestations that develop. The mechanisms involved in this differential responsiveness appear to reflect different T-cell cytokine response patterns. Asymptomatic patients with the clinically silent presentation of ‘asymptomatic microfilaraemia’, who have been previously described as being ‘immunosuppressed’ with respect to their generating pro-inflammatory (Th1-type) immune responses to parasite antigen, are now recognized to be fully responsive to parasite antigen but to produce cytokines and mediators that have primarily anti-inflammatory (Th2-like) effects. Studies with immunodeficient mice have indicated the existence of two alternative pathways to the development of lymphatic pathology: one dependent on the induction of inflammatory reactions by the host immune response, the other entirely independent of the immune system and reflecting the direct actions of the parasite or its products on the lymphatics. As histopathology of affected human lymphatics is consistent with this hypothesis, it may be that the lymphatic pathology seen normally in the amicrofilaraemic, highly immunoresponsive infected patients derives from inflammation induced by immune responses to parasite antigen, whereas the lymphatic pathology sometimes seen coexisting with the ‘immunosuppressed’ state of asymptomatic microfilaraemia actually reflects lymphatic damage that is not immunologically mediated. Though little information exists about the ‘natural history’ of lymphatic filariasis, there is no evidence for an inevitable progression from one clinical form to another. Instead, there appears to be a definite plasticity in the response that depends on prior (? pre-natal) and current exposure to the parasite as well as on the immunomodulatory effects it induces. This plasticity does not appear to be complete, however, as there is no evidence that a chronically infected host who has developed strong pro-inflammatory immune responses can subsequently become sufficiently ‘down-regulated’ to support an asymptomatic microfilaraemia type of infection. Another possible constraint to the plasticity of the clinical and immunological responses may be the genetic determination of certain unusual syndromes, such as tropical pulmonary eosinophilia or TPE, though this hypothesis remains to be proven.

A major question in the study of any parasitic disease is the relationship between infection and clinical disease. The public health importance of lymphatic filariasis has generated a large body of research in this area, both in laboratory studies (Ottesen, 1984,1989; Maizels & Lawrence, 1991) and in the field (Hayashi, 1962; Hairston & Jachowski, 1968; Denham & McGreevy, 1977; Vanamail et al. 1989 b; Bundy, Grenfell & Rajagopalan, 1991; Srividya et al. 1991 b). Despite this, there is still no conclusive explanation for the apparently complex relationship between infection and clinical disease observed in human communities. At least part of the problem may lie in the current impossibility of measuring adult worm burden in vivo (Pichon et al. 1980; Denham & Fletcher, 1987; Das et al. 1990; Grenfell et al. 1990). Although there has recently been significant progress in the development of immuno logical markers for infection status in humans (Ottesen, 1989; Day et al. 1991 a), microfilaraemia is still the most reliable measure of current infection in the field. Studies in endemic areas indicate that, far from there being any simple direct relationship between microfilaraemia and disease status, it is possible to find some individuals with microfilariae in their blood but no disease, and indeed with all other combinations of infection and disease status (Hairston & de-Meillon, 1968; Hairston & Jachowski, 1968; Beaver, 1970; Bryan & Southgate, 1976; Denham & McGreevy, 1977; Pani et al. 1991). Furthermore, the proportions of people in different categories are often observed to vary between endemic areas (Denham & McGreevy, 1977; Day et al. 1991 a).

The Dactylogyrus fauna was studied from the gills of 293 roach between February 1988 and April 1989. Roach were caught from three interconnected lakes in Central Finland. Nine Dactylogyrus species were found. Of these the seven most abundant were used for studying the abundance of species on different gill-arches, niche breadth and niche overlap between the species. It was found that all species had a species-specific preference for certain gill-arches. The total abundances of all the species went through distinct seasonal changes, peaking during the spring and early summer. Some seasonal variation was also exhibited in gill-arch preferences. Niche breadth and species overlap were mostly higher during the period of peak occurrence of the various species, but in the case of some species they appeared to be unrelated to abundance, indicating possible niche restriction by other species. Most significant was the restriction of D. micracanthus to the fourth gill-arch during the period of peak occurrence of D. similis. Experimental work is needed to verify the reasons for variations in the microhabitat distribution of certain Dactylogyrus species.

Two clones from separate isolates of Trypanosoma evansi in Indonesia were found by polymerase chain reaction (PCR) analyses to contain 3 different repeated nuclear DNA sequences of Trypanosoma brucei spp: the consensus sequence for a highly repetitive 177 base pairs and the gene repeats encoding procyclin and the spliced leader. In addition, the 994 bp minicircle sequence of one of the clones was determined, and PCR amplification primers specific for minicircles of T. evansi were identified that do not amplify minicircle sequences in the T. brucei spp. clones tested.

The prevalence of Schistosoma mansoni infections of Biomphalaria pfeifferi shows seasonal variation. Field data from Zimbabwe show annual ranges from 0 to 7%. In this paper a mathematical model of B. pfeifferi population dynamics and S. mansoni epidemiology is used as a framework for analysis of these patterns. Snail fecundity is a function of snail age and of temperature, and is apparently affected by other seasonal factors. The pre-patent period is dependent on temperature. Infection affects snail fecundity and mortality. Parameter values are derived from previous field and laboratory studies. The force-of-infection is estimated from the analysis of size-prevalence data. Using observed temperatures, model output agrees well with field data on snail abundance and prevalence of infection over a 14-month period. Seasonal variation in prevalence largely reflects variation in the pre-patent period and in snail population age structure. The possible role of seasonality in the force-of-infection is discussed. Prevalence patterns are not greatly affected by year-to-year differences in temperature. Significant seasonal variation in snail–man transmission rates is expected.

The mechanism by which small animals such as rodents resist or eliminate nematode parasites requires mucosal in flammation as the final effector of the immune response. The resulting freedom from chronic infection may be worth the price of short-term illness. Putative vaccines which attempt to enhance the natural effect will have to take into account the inflammatory cost to the host. Human helminthiases involve a more stable equilibrium between host and parasite. The medical literature on hookworm disease and clinical ascariasis describes, for the former, some chronic inflammatory effects correlated with worm burden, but for the latter a less quantified or predictable set of detrimental effects. We describe a current, systematic study of the inflammatory response to whipworm infection, in which anaemia, growth retardation and intestinal leakiness are viewed as predictable consequences related to infection intensity. There is evidence for the absence of cell-mediated immunopathology. However, a specific, IgE-mediated local anaphylaxis may, at least partly, mediate the deleterious effects. Increased numbers of mucosal macrophages may also contribute to the chronic, systemic effects through their output of cytokines. Similar attempts to show the mechanisms of pathogenesis and quantify the effects of hookworm disease should be undertaken.

Pseudodiplorchis americanus infects the urinary bladder of a desert toad and feeds exclusively on blood, producing the iron-rich waste product haematin. The host, Scaphiopus couchii, retains dilute urine within the urinary bladder as a water store throughout hibernation and parasite waste will therefore not be eliminated. This study utilized atomic absorption spectrophotometry to quantify the iron within the urinary bladders of infected and uninfected toads after different periods of hibernation. During the first 4 months hibernation, no detectable iron, above that of the controls, was found in the urine and bladder tissue of infected animals, but after 5–6 months hibernation there was a small but not significant increase. The iron contained within most individual parasites was greater than that detected in the urine and bladder tissue of the host, and the total iron in each parasite infrapopulation was 3–83 times greater than the urinary bladder iron. This suggests that the parasites do not regurgitate their gut contents during host hibernation. The amount of iron in individual parasites increased with time after migration to the bladder and this was used to estimate the amount of blood ingested by P. americanus. At a controlled temperature of 25 °C, the estimated rate of ingestion increased from 0·33 μl blood/parasite/week by worms 2 weeks post-migration to a maximum of around 1·6 μl by worms 5 months post-migration. The rate of blood ingestion by older worms was also calculated as a function of the hibernation period (since this did not correspond to worm age) assuming complete gut evacuation at the start of hibernation. This provided maximum estimates of ingestion rate of 1·9–5·3 μl blood/parasite/week.

Numerical analyses of Trypanozoon taxonomy are presented, based on the isoenzyme data of Stevens et al. (1992). The previous study used a reduced range of enzymes compared with earlier work; the analyses indicate the value of this rationalized system. Both recently isolated trypanosome stocks and previously studied populations were included, allowing detailed comparison with earlier studies. Relationships between zymodemes were calculated with an improved similarity coefficient program, using Jaccard's coefficient (1908), and by Nei's method (1972). Dendrograms were constructed from the matrices produced with the group-average method. The groupings produced by both numerical methods were in close agreement, and the clusters of related principal zymodemes largely matched the species, subspecies and strain groups proposed by previous workers. Trypanozoon biochemical taxonomy is reviewed and the groupings reinforced by this study are: the mainly East African strain groups, busoga, zambezi, kakumbi, kiboko and sindo; T.b. gambiense and the bouaflé strain group from West Africa, and T. evansi; an intermediate bouaflé/busoga group was also recognized.

Faecal egg counts are often used to measure Schistosoma mansoni infection, but the considerable variation between successive counts complicates their interpretation. The stochastic model described in this paper gives a description of observed egg counts in a population and can be used as a tool to gain an insight into the underlying worm load distribution. The model distinguishes between two sources of variation in egg counts: (1) variation caused by the difference in worm load between individuals, and (2) the variability of egg counts for an individual with a given worm load. Empirical data, single and repeated measurements, from surveys in five villages in Burundi and Zaire have been used to fit and validate the model. We have discussed possible mechanisms that explain the differences in estimated values between the villages. The model indicates that the expected number of eggs in a stool sample per S. mansoni worm pair is lower than suggested by autopsy data and that, possibly as a consequence of immunity, the inter-individual variation in worm loads decreases with age.