Resistance in Fasciola hepatica to triclabendazole (‘Fasinex’) has emerged in several countries. Benzimidazole resistance in parasitic nematodes has been linked to a single amino acid substitution (phenylalanine to tyrosine) at position 200 on the β-tubulin molecule. Sequencing of β-tubulin cDNAs from triclabendazole-susceptible and triclabendazole-resistant flukes revealed no amino acid differences between their respective primary amino acid sequences. In order to investigate the mechanism of triclabendazole resistance, triclabendazole-susceptible and triclabendazole-resistant flukes were incubated in vitro with triclabendazole sulphoxide (50 μg/ml). Scanning and transmission electron microscopy revealed extensive damage to the tegument of triclabendazole-susceptible F. hepatica, whereas triclabendazole-resistant flukes showed only localized and relatively minor disruption of the tegument covering the spines. Immunocytochemical studies, using an anti-tubulin antibody, showed that tubulin organization was disrupted in the tegument of triclabendazole-susceptible flukes. No such disruption was evident in triclabendazole-resistant F. hepatica. The significance of these findings is discussed with regard to the mechanism of triclabendazole resistance in F. hepatica.

Data on parasites of rodents, collected over an 18-year period on the Iberian peninsula, were used to find the determinants of parasite species richness. A total of 77 species of helminth parasites (nematodes, cestodes and digeneans) was identified among 16 species of rodents. Parasites were classified into groups according to their specificity towards their host and their life-cycle. A working phylogeny of the rodents was proposed on the basis of molecular and paleontological data and for each host the following parameters were recorded: sample size, weight, geographical range, longevity, and life-style. Two comparative methods were used, the independent comparisons method of Pagel (1992) and the distance matrix method of Legendre, Lapointe & Casgrain (1995). The second method has the advantage of measuring the relative contribution of phylogeny. Both methods gave similar results. Overall parasite species richness correlated only with host sample size. Host body size does not correlate with any subset of parasite species richness. However, host phylogeny is a good predicator of specific parasites and the species richness of digeneans correlates with host geographical range. A phylogenetic reconstruction of host relations was performed using the parasites belonging to subgroups in which richness is correlated with host phylogeny. These parasite species were treated as Dollo characters, i.e. we made the assumption that the loss of a parasite species is irreversible. The consensus tree obtained reflects the major phylogenetic divisions of the host group. Finally, this study illustrates the relative importance of processes acting at different temporal and spatial scales (evolutionary time and actual geographical range of hosts) in determining the structure of helminth parasite fauna.

The Consortium for Anthelmintic Resistance and Susceptibility (CARS) brings together researchers worldwide, with a focus of advancing knowledge of resistance and providing information on detection methods and treatment strategies. Advances in this field suggest mechanisms and features of resistance that are shared among different classes of anthelmintic. Benzimidazole resistance is characterized by specific amino acid substitutions in beta-tubulin. If present, these substitutions increase in frequency upon drug treatment and lead to treatment failure. In the laboratory, sequence substitutions in ion-channels can contribute to macrocyclic lactone resistance, but there is little evidence that they are significant in the field. Changes in gene expression are associated with resistance to several different classes of anthelmintic. Increased P-glycoprotein expression may prevent drug access to its site of action. Decreased expression of ion-channel subunits and the loss of specific receptors may remove the drug target. Tools for the identification and genetic analysis of parasitic nematodes and a new online database will help to coordinate research efforts in this area. Resistance may result from a loss of sensitivity as well as the appearance of resistance. A focus on the presence of anthelmintic susceptibility may be as important as the detection of resistance.

The nuclear DNAs of 71 trypanosome stocks from different African countries, representative of the three Trypanosoma brucei subspecies, and one T. evansi stock, have been analysed by the combined use of restriction endonuclease digestion, gel electrophoresis and molecular hybridization with both trypanosome surface-antigen-specific and undefined genomic DNA probes. In contrast with T. brucei brucei and T. brucei rhodesiense stocks, all the T. b. gambiense stocks are characterized by a conserved, specific DNA band pattern, regardless of the probe. This allows T. b. gambiense to be non-ambiguously identified. On the contrary, T. b. brucei and T. b. rhodesiense, which could not be discriminated by the same criteria, both yield highly variable DNA band patterns. Our data confirm that domestic animals like pig, dog and sheep constitute a potential reservoir for T. b. gambiense. Using a numerical analysis of the DNA hybridization patterns we have measured the degree of similarity between the 72 trypanosome stocks. This investigation shows that all T. b. gambiense stocks are included in the same homogeneous population, while the stocks from the two other subspecies seem to be distributed in several heterogeneous groups, some of these showing correlation with the geographical origin of the trypanosomes. It is concluded that (i) T. b. gambiense stands out as a real subspecies that has undergone a distinct evolution relative to the ‘non-gambiense’ group, (ii) the alleged T. b. rhodesiense subspecies does not fit with any of the groups evidenced by our cladistic analysis and hence does not appear as a distinct subspecies and (iii) ‘non-gambiense’ trypanosomes are probably evolving much more rapidly than T. b. gambiense. Different aspects of trypanosome relationships and evolution are discussed.

According to the World Health Organization the global estimate of malaria is over 200 million infections, the majority of which are caused by the most life-threatening species, Plasmodium falciparum (Report of the Steering Committees of the Scientific Working Groups on Malaria, World Health Organization, June 1983). The causative agent of the disease, the malarial parasite, requires two hosts: a blood-sucking mosquito and a blood-containing vertebrate. Commonly, infection of the vertebrate begins when an infected mosquito bites a suitable vertebrate and injects minute sporozoites into the bloodstream. Within 30 mm the introduced sporozoites leave the bloodstream and enter parenchymal cells of the liver (mammals) or endothelial cells (birds). In these sites the parasite undergoes asexual multiplication (= exo-erythrocytic schizogony) producing daughter progeny called merozoites. The exo-erythrocytic merozoites are released from the tissues into the circulation where they invade red blood cells. Within an erythrocyte the merozoite undergoes asexual multiplication (= erythrocytic schizogony) producing a substantial number of merozoites. The erythrocyte lyses, merozoites are released, and invasion of another erythrocyte may then take place. The synchronous rupture of the red cell and merozoite release is marked by the periodic fever–chill cycles so characteristic of the malarial infection. Some merozoites continue to reinvade other erythrocytes and multiply by asexual means, whereas others enter erythrocytes and differentiate into sexual stages, male or female gametocytes. When a suitable mosquito feeds on an infected vertebrate gametocytes are ingested and the sexual cycle of development is initiated. In the mosquito stomach the gametocytes transform into gametes, fertilization takes place, the resultant worm-like zygote penetrates the cells of the mosquito gut and comes to lie on the outer surface of the stomach. Here each zygote forms a cyst-like body, the oocyst, within which thousands of sporozoites are produced by asexual multiplication. When the swollen oocysts burst, sporozoites are freed and these make their way to the salivary gland. At the next blood feeding the mosquito injects the infective sporozoites and the life-cycle is completed.

The frequency of recombination between unlike genotypes is central to understanding the generation of genetic diversity in natural populations of malaria. Here we suggest a way of investigating the problem which could complement conventional biochemical approaches to the population genetics of malaria. Sex allocation theory is one of the most successful areas of evolutionary biology. A well-supported prediction is that progressively less female-biased sex ratios are favoured with more outcrossing; equal numbers of males and females being evolutionarily stable in randomly mating outbred populations. We present a simple game theory model to support the idea that outcrossing rates in malaria will be correlated with the sex ratio of gametocytes in the peripheral blood of vertebrate hosts. Blood films from epidemiological surveys and culture-adapted isolates from Madang Province, Papua New Guinea, were used to estimate average gametocyte sex ratio of Plasmodium falciparum in the area. The geometric mean proportion of males in the population was 0.18 (95% confidence limits: 0.15–0.22). From our model, we estimate that, on average, 36% of zygotes are the result of outcrossing. This estimate assumes that most microgametes released following exflagellation are capable of fertilization. If, on average, fewer than about 70% of microgametes are capable of fertilization (as is the case in at least one other species of Plasmodium), the observed sex ratio would be consistent with between zero and 36% of zygotes being the result of outcrossing. These estimates suggest that there is usually a numerically dominant genotype in the gametocyte population in a blood meal, and that a considerable amount of selfing is occurring in P. falciparum populations in the Madang region, even though it is an area of intense year-round transmission.

Recent research suggests that vertical transmission may play an important role in sustaining Toxoplasma gondii infection in some species. We report here that congenital transmission occurs at consistently high levels in pedigree Charollais and outbred sheep flocks sampled over a 3-year period. Overall rates of transmission per pregnancy determined by PCR based diagnosis, were consistent over time in a commercial sheep flock (69%) and in sympatric (60%) and allopatric (41%) populations of Charollais sheep. The result of this was that 53·7% of lambs were acquiring an infection prior to birth: 46·4% of live lambs and 90·0% of dead lambs (in agreement with the association made between T. gondii and abortion). No significant differences were observed between lamb sexes. Although we cannot distinguish between congenital transmission occurring due to primary infection at pregnancy or reactivation of chronic infection during pregnancy, our observations of consistently high levels of congenital transmission over successive lambings favour the latter.

Both histological and histochemical studies have been made of the food, mode of feeding, gut cell structure and digestive sequences in a selected number of digenetic trematodes.

The species investigated range from gut-dwelling trematodes feeding on the superficial epithelial tissues and associated mucoid secretions of the host to those forms living within the respiratory and circulatory system and feeding exclusively on blood.

The mode of feeding is suctorial, brought about by the muscular pharynx and normal attachment process of the oral sucker. In one case there is evidence that this purely mechanical process is supplemented with enzymic secretions produced by the trematode which have a histolytic effect upon host tissues.

On the basis of gut cell structure the species investigated are divided into those with cells and associated microvilli that vary in size and appearance so that the gastrodermal border is irregular in outline, and those with a gastrodermis comprising regular cuboidal or columnar cells bearing microvilli organized into a striated border. There is evidence to suggest that this difference in gut cell structure reflects differences in digestion. In all cases, gland cells are absent from the gastrodermis which is capable of both secretory and absorptive functions.

Digestion is predominantly an extracellular process, but the exact sequence varies according to the nature of the food ingested and different degrees of adaptation are shown by the trematodes to the blood-feeding habit. The tissue-feeding species show less modification.

I wish to thank Dr J. B. Jennings for helpful discussion and advice throughout the course of this work. The study was undertaken during the tenure of a Studentship from the Department of Scientific and Industrial Research.

The amastigote form of the human pathogen Leishmania mexicana contains high proteinase activity, some 20 times greater than that in the promastigote form and macrophages and appreciably higher than the activity in other flagellate protozoa. The main amastigote enzymes are soluble, whereas those of the promastigote are particulate, and have inhibitor sensitivities characteristic of cysteine proteinases. The very high soluble proteinase activity of L. mexicana amastigotes may be a primary factor in the survival and growth of this mammalian stage in its potentially degradative intracellular habitat.

Life history of Ixodes ricinus.

1. Under laboratory conditions, at 60–70° F. the pre-oviposition period was usually found to last 15–22 days.

2. Eggs began to hatch after about 4 weeks at 60–70° F. and after 37 weeks at 50–60° F. in the laboratory.

3. Unfed larvae placed on a hedgehog mostly dropped off gorged in 88–120 hours (shortest feeding period 67½ hours). Larvae gorged on sheep in 65 hours.

4. From larvae gorged in August and kept in the laboratory, nymphs emerged after 6–7 weeks. From larvae gorged in September and October, nymphs emerged after 28–37 weeks.

5. Unfed nymphs gorged in 4–7 days, mostly in 5 days.

6. From nymphs gorged on sheep in spring and summer, and kept in the laboratory at 60–70° F., adults emerged after 10–19 weeks.

7. Females gorged on sheep in 7–11 days, mostly in 8–9 days.

8. Sex ratio: about 5: 9 (basis: 362 males and 671 females emerged from nymphs in the laboratory).

9. Longevity unfed: males and females survived 21 months; one male survived for 31 months in the laboratory. Larvae survived 2 years; some were able to feed after fasting 15 months, and nymphs after fasting 13 months.

Bionomics.

10. Hosts: one or more stages of I. ricinus have been found on the following hosts. Mammals: horse, dog, fox, hedgehog, hare, rabbit, stoat. Birds: hoodie-crow, rook, jackdaw, starling, golden plover, barn owl, kestrel hawk, and merlin hawk.

11. When Blackface and Cheviot sheep were kept on the same pastures in spring, the former became more heavily infested with ticks than the latter. It is said by shepherds that ewes weakened by lambing are more liable to infestation than strong barren ewes. Comparative counts of the infestations of a barren ewe and two ewes with lamb appear to confirm the shepherds' view. Non-acclimatised (experimental) sheep, when exposed on a tick infested farm, rapidly. lost condition at first, and subsequently began to recover. When these sheep, whose health and general condition were low, were compared with native sheep their average infestation with ticks was found to be higher than that of the natives. As the health and weight of the experimental sheep improved, the average infestations of these and of the native stock approximated.

12. Reports on the geographical distribution of I. ricinus in various districts in Scotland, received from sheep farmers and others, are recorded.

13. There is a distinct seasonal periodicity in the activity of I. ricinus. It appeared on sheep early in March, and became increasingly numerous during April-May. In June the numbers began to decrease, and steadily diminished until late August, when there occurred a slight increase in average infestation of sheep. This increase was of short duration and by the middle of November no ticks were found on sheep. The same periodicity was observed in the following spring and summer. An occasional tick was found on sheep during the winter, in mild warm weather. Adult ticks first appeared in early March. Nymphs appeared by the middle of March. Larvae were apparently most numerous in May–June.

14. The seasonal incidence of I. ricinus on sheep in 1930 bore no apparent relation to rainfall.

15. When the variations in average weekly air temperature for 1930–31 are charted in relation to the seasonal incidence of ticks on sheep for that period and the curves compared, there seems to be a correlation between temperature and tick activity. The data indicate that the tick infests sheep in the greatest numbers at 45–60° F. The lower critical temperature for tick activity in nature appears to be 40–45° F., and the higher 60–70° F.

16. Over a thousand gorged nymphs and many larvae have been collected in the field and bred out in the laboratory; in none of them has there been any evidence of hyperparasitism.

Circumstantial evidence suggests that the earlier detachment of Boophilus microplus larvae from highly resistant cattle follows the release of histamine at the attachment site. In vivo and in vitro experiments show that a proportion of the larvae will detach following injection or infusion of histamine. Other mediators such as bradykinin, prostaglandin E2, 5-hydroxytryptamine and dopamine have little or no effect on tick behaviour in vivo. Sensitivity to histamine declines as larval attachments stabilize, and repeated injections have no effect on the weight of larvae after 3 days on the host. Response to histamine is discussed in relation to host resistance, histology of the feeding lesion and larval behaviour.

Planning of the control of Plasmodium falciparum malaria leads to a need for models of malaria epidemiology that provide realistic quantitative prediction of likely epidemiological outcomes of a wide range of control strategies. Predictions of the effects of control often ignore medium- and long-term dynamics. The complexities of the Plasmodium life-cycle, and of within-host dynamics, limit the applicability of conventional deterministic malaria models. We use individual-based stochastic simulations of malaria epidemiology to predict the impacts of interventions on infection, morbidity, mortality, health services use and costs. Individual infections are simulated by stochastic series of parasite densities, and naturally acquired immunity acts by reducing densities. Morbidity and mortality risks, and infectiousness to vectors, depend on parasite densities. The simulated infections are nested within simulations of individuals in human populations, and linked to models of interventions and health systems. We use numerous field datasets to optimise parameter estimates. By using a volunteer computing system we obtain the enormous computational power required for model fitting, sensitivity analysis, and exploration of many different intervention strategies. The project thus provides a general platform for comparing, fitting, and evaluating different model structures, and for quantitative prediction of effects of different interventions and integrated control programmes.

Bloodstream forms of Trypanosoma congolense (2 clones: ILNat3·1 and IL3000, and 4 stocks: IL2079, IL2466, IL3266 and CP-81) were continuously cultivated in vitro at 34–36 °C in the absence of feeder cell layers, using HMI-93 medium which was modified from Iscove's modified Dulbecco's MEM (Flow Laboratories, Irvine, Scotland). The modification was done by supplementing the medium with 0·05 mM bathocuproine sulphonate, 1·5 mM L-cysteine, 0·5 mM hypoxanthine, 0·12 mM 2-mercaptoethanol, 1 mM sodium pyruvate, 0·16 mM thymidine, 20% (v/v) heat-inactivated young goat serum and 5% (v/v) Serum Plus™ (Hazleton Biologics, Lenaxa, KS, USA). Trypanosomes obtained from two different sources were used to initiate primary cultures: (1) metacyclic forms which were produced in vitro at 27 °C, and (2) bloodstream forms obtained from Balb/c mice which had been infected with the bloodstream forms transformed in vitro from the metacyclic forms. Metacyclic forms placed in 25 cm2 T-type (T-25) flasks rapidly attached to the bottom of the flasks and transformed to bloodstream forms during the initial 24 h and continued to proliferate. The bloodstream forms isolated from the infected mouse blood by means of diethylaminoethyl cellulose (DE52) column chromatography also continued to proliferate in the flasks. Cultures were maintained by replacing the medium every 24 h. Every 4–5 days, the attached bloodstream forms were resuspended in fresh medium by gentle pipetting and then were subcultured. The method was further simplified by initiating primary cultures directly with 10 μl of the tail blood of infected mice in 24-well culture plates and then by subcultivating either in wells or in T-25 flasks. The shortest population doubling time, 9 h, was achieved by seeding subcultures with 106 bloodstream forms/ml. The bloodstream forms propagated in this system were morphologically similar to those seen in infected mouse blood, they were covered with a surface coat as examined by electron microscopy and they were infective to mice.

The four most important forms of malnutrition worldwide (protein-energy malnutrition, iron deficiency and anaemias (IDA), vitamin A deficiency (VAD), and iodine deficiency disorders (IDD)) are examined below in terms of their global and regional prevalences, the age and gender groups most affected, their clinical and public health consequences, and, especially, the recent progress in country and regional quantitation and control. Zinc deficiency, with its accompanying diminished host resistance and increased susceptibility to infections, is also reviewed. WHO estimates that malnutrition (underweight) was associated with over half of all child deaths in developing countries in 1995. The prevalence of stunting in developing countries is expected to decline from 36% in 1995 to 32·5% in 2000; the numbers of children affected (excluding China) are expected to decrease from 196·59millions to 181·92millions. Stunting affects 48% of children in South Central Asia, 48% in Eastern Africa, 38% in South Eastern Asia, and 13–24% in Latin America. IDA affects about 43% of women and 34% of men in developing countries and usually is most serious in pregnant women and children, though non-pregnant women, the elderly, and men in hookworm-endemic areas also comprise groups at risk. Clinical VAD affects at least 2·80million preschool children in over 60 countries, and subclinical VAD is considered a problem for at least 251millions; school-age children and pregnant women are also affected. Globally about 740million people are affected by goitre, and over two billions are considered at risk of IDD. However, mandatory salt iodisation in the last decade in many regions has decreased dramatically the percentage of the population at risk. Two recent major advances in understanding the global importance of malnutrition are (1) the data of 53 countries that links protein-energy malnutrition (assessed by underweight) directly to increased child mortality rates, and (2) the outcome in 6 of 8 large vitamin A supplementation trials showing decreases of 20–50% in child mortality.

The term ‘shotgun metagenomics’ is applied to the direct sequencing of DNA extracted from a sample without culture or target-specific amplification or capture. In diagnostic metagenomics, this approach is applied to clinical samples in the hope of detecting and characterizing pathogens. Here, I provide a conceptual overview, before reviewing several recent promising proof-of-principle applications of metagenomics in virus discovery, analysis of outbreaks and detection of pathogens in contemporary and historical samples. I also evaluate future prospects for diagnostic metagenomics in the light of relentless improvements in sequencing technologies.

An epidemiological survey of intestinal helminthiases was conducted on 766 primary school children aged 5–16 years from Ile-Ife, Nigeria. On the basis of stool examinations, the prevalence of Ascaris lumbricoides, Trichuris trichiura, hookworm and Strongyloides stercoralis was 88·5, 84·5, 33·1 and 3% respectively. Intensity of infection was measured indirectly by egg counts for each species of helminth and also by counting worms passed after chemotherapy in the case of A. lumbricoides. The influence of host age and sex on infection levels was assessed. Relationships between the intensities of A. lumbricoides, T. trichiura and hookworm in individual children were identified. After anthelmintic treatment with levamisole, the frequency distribution of A. lumbricoides per host and the relationship between parasite fecundity and worm burden were investigated.

Host immunoglobulin G (IgG) crossed the gut wall into the haemocoel of adult Rhipicephalus appendiculatus female ticks when they fed on guinea-pigs. Guinea-pig IgG was also found in saliva of the feeding ticks. The concentration and antibody activity of IgG in haemolymph, salivary gland extract (SGE) and saliva at different stages of tick feeding were detected by enzyme-linked immunoassay. Specific activity of the IgG in tick samples was determined by feeding ticks on guinea-pigs which were immunized with killed Escherichia coli: 35–42% of the antibody activity in guinea-pig immune serum remained in the tick samples. The high relative concentration of IgG in tick saliva at later stages of feeding suggests that the tick may have a mechanism for getting rid of foreign proteins via the salivary gland. Such a mechanism could involve lgG binding proteins (1GBPs) which were found in both haemolymph and SGE of female ticks at day 6 of feeding using a guinea-pig IgG–agarose affinity column. In female ticks, the Mr of 1GBPs in SGE (23 and 57 kDa) were less than those in haemolymph (78 and > 100 kDa). The existence of 1GBPs in both the tick salivary gland and haemolymph indicate that haemolymph and salivary gland cooperate to remove foreign proteins, e.g. host immunoglobulin, from the body during feeding. This mechanism may be a part of the tick self-defence system.

American trypanosamiasis occurs in nature as a sylvatic cycle, where Trypanosoma cruzi interacts with wild triatomines and mammalian reservoirs, such as marsupials, rodents, armadillos and other animals. Due to difficulties in trying to isolate T. cruzi stocks from the sylvatic cycle, very few studies have been performed in order to understand the parasite infection in natural environments. Traditionally T. cruzi has been considered to be composed of a highly heterogeneous population of parasites. In contrast, the mini-exon and the 24Sα rRNA gene loci have shown that T. cruzi stocks can be clustered in 2 major phylogenetic groups: lineage 1 and lineage 2. In this report, 68 recently isolated T. cruzi samples from the sylvatic cycle belonging to different geographical areas in Rio de Janeiro, Brazil, have been typed based on a variable spot in the non-transcribed spacer of the mini-exon gene. Eight isolates were from triatomines, 26 stocks were from golden-lion tamarins, 31 from opossums, 2 from rodents and 1 from a three-toed sloth. Thirty (44% – 30/68) isolates were typed as lineage 1, while 36 (53% – 36/68) isolates were typed as lineage 2. Two opossums presented mixed infection. Therefore, 3% (2/68) of the isolates were typed as lineage 1+lineage 2. Using these geographical regions as models of sylvatic environments, it was observed that 96% of the Didelphis marsupialis were infected by lineage 2 isolates, while all 26 golden-lion tamarins were infected by lineage 1. The results show preferential association of the 2 lineages of T. cruzi with different hosts, composing the complexity of the sylvatic cycle.

Rodents are shared intermediate or paratenic hosts for Echinococcus multilocularis, Toxocara spp. and Toxoplasma gondii, and may serve as valuable indicators for assessing the occurrence and the level of environmental contamination and infection pressure with free-living stages of these zoonotic parasites. We investigated 658 non-commensal rodents for parasite infections in the canton of Geneva, Switzerland. The prevalence of infection with E. multilocularis was highest in Arvicola terrestris captured in the north-western area (16·5%, CI: 10·1%–24·8%), possibly reflecting a higher red fox density due to the low incidence of sarcoptic mange in this part of the canton. The exposure rate to Toxocara spp. was highest in the urban area (13·2%, CI: 7·9%–20·3%), and may account for higher densities of domestic carnivore and red fox definitive hosts within the city. Exposure to T. gondii was widespread (5·0%, CI: 3·2–7·4%), indicating a ubiquitous distribution of infected cat definitive hosts. Interestingly, a widespread distribution of Taenia taeniaeformis, a parasite mainly transmitted by cats, was similarly evidenced in A. terrestris. Distinct spatial patterns for the different zoonotic parasites likely reflected differences in distribution, abundance, and habitat use of the respective definitive hosts. These results highlight the potential value of rodents as shared indicators for these pathogens.

New or improved techniques for recovering Schistosoma mansoni from the skin, lungs and liver have enabled us to trace the attrition of a challenge infection in naive (i.e. previously uninfected) and chronically infected mice. Within each experiment, the numbers of schistosomes recovered from the skin of naive mice on day 2 after challenge or from the skin and lungs on days 3, 4 or 5, did not differ significantly from the numbers recovered from the liver on days 14, 21, 28 or 35. Approximately 65% of cercariae which penetrated the skin failed to be recovered from naive mice by any of the assays and it appeared that these schistosomes had already died in the skin in the first 24 h. No further significant loss of the infection was detected in naive mice. In chronically infected mice a further attrition of the challenge infection was demonstrated in two distinct phases. An ‘early phase’ occurred within the first 3 days of exposure and accounted for the death of 30% of the remaining parasites. A ‘late phase’ occurred between days 6 and 14 and accounted for an additional 43% of deaths. Thus, the two phases of attrition accounted for a loss of approximately 73% of the infection that would have survived in naive mice. The late phase of attrition could be demonstrated before the primary infection had matured, in contrast to the early phase of attrition which was seen only after egg laying had commenced. We believe that the early phase of attrition takes place in the skin and the late phase occurs after the schistosomes have left the lungs, either en route for the liver or as soon as they arrive in that organ. The results suggest that there are two distinct mechanisms of immunity against re-infection with S. mansoni in mice.