Specific sequences derived from the gene for the precursor to the major merozoite surface antigens (PMMSA) of Plasmodium falciparum have been expressed in Escherichia coli and the products have been used to produce antibodies. These antibodies, together with monoclonal antibodies, have been used to investigate the form of the PMMSA protein associated with merozoites. Polypeptide fragments derived by processing from the PMMSA protein have been detected in extracts of merozoites and assigned to locations within the PMMSA coding sequence.

An enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies to Encephalitozoon cuniculi in man, using spores derived from tissue culture as antigen. Negligible cross-reactions were found with other microsporidia of vertebrate or of invertebrate origin and there was no cross-reaction with Toxoplasma gondii, using ELISA, immunoperoxidase or immunofluorescence staining. A high prevalence of antibodies to E. cuniculi was found in patients suffering from schistosomiasis, malaria and neurological and psychiatric disorders, but not in healthy individuals.

The first two phases in the growth of Maritrema linguilla in Ligia oceanica, which occur in unencysted specimens in the pleopod sinuses, from day 0 to day 28 post-infection (p.i.), include a 3-fold increase in size, an enormous increase in the relative size of the excretory bladder, metamorphosis of papilliform extensions of the tegument into microvilli and development of single pointed spines and cystogenous gland cells. The third phase, from day 29 to day 38 p.i., following migration to the dorsal haemocoel, includes production of 3 cyst wall layers, the loss of single pointed spines and a considerable decrease in the relative size of the excretory bladder. The fourth phase from day 39 to day 42 p.i., involves a further increase in body size and the formation of digitiform spines. The fifth, sixth and final phases from day 43 to day 120 p.i. first include the completion of the last two cyst wall layers and then an increase in body size and development of the digestive, excretory and reproductive systems to almost adult dimensions but sperm, ova, vitelline globules and eggs are produced only in the final host.

The tegumental membrane potential (–63±2·9 mV) of adult male Schistosoma mansoni in RPMI-1640 is significantly depolarized (–26±7·3 mV) when the parasite is incubated in inorganic media (Hank's Balanced Saline or RPMI-1640 without organic constituents). Of 9 amino acids (L-glutamine, D-glutamine, L-arginine, L-proline, L-aspartate, L-glutamate, L-asparagine, L-isoleucine and L-methionine) L-glutamine alone is sufficient to repolarize the membrane potential to a value (–56±4·5 mV) not significantly different from that found in RPMI-1640. Repolarization by glutamine is dose-dependent, with significant effects obtained as low as 0·10 mM. The concentration of phosphate in the medium also significantly alters the membrane potential. Physiological levels of phosphate (5·6 mM) are necessary in conjunction with L-glutamine to obtain the full repolarization of the membrane potential. In the absence of organic constituents, the membrane potential is strongly dependent on the external medium pH. When L-glutamine is present in the medium, the membrane potential becomes virtually independent of the external pH.

The long-term, in vivo effects of a single, subcurative dose (200 mg/kg body weight of mouse) of praziquantel on the structure of adult Schistosoma mansoni and on the process and speed of tegumental repair are described. In both male and female worms praziquantel caused often extensive damage to the tegument, in the form of surface blebbings, swellings and lesions, and vacuolization and disruption of the subtegumental tissues. Repair of the drug-induced tegumental damage occurred slowly with partial and, more rarely, complete repair only being seen after 65 days post-treatment (p.t.), although signs of damage were still observed, particularly in male worms, at 100 days p.t. In contrast, repair of damage to the subtegumental/parenchymal tissues including the tegumental perikarya occurred relatively quickly, with the majority of worms examined appearing normal by 8–12 days p.t. The possible role(s) of the host immune response in relation to the speed of tegumental repair in vivo is discussed.

Antisera were produced by immunizing rabbits with either a trichloroacetic acid-soluble fraction, or a high molecular weight (Mr) fraction of Schistosoma mansoni SEA (a salinesoluble fraction of homogenized egg). Both of these sera reacted monospecifically in immunoelectrophoresis against unfractionated SEA, recognizing a cathodally migrating antigen. This antigen had been identified previously as being responded to by S. mansoni-infected mouse sera, and has been designated K3 (Kappa 3). The rabbit antisera were used to partially characterize antigen K3 as having Mr in the range > 750 – 70 K, and being resistant to boiling, resistant to the action of proteases, but sensitive to periodate. It partially binds to Concanavalin A. In addition to SEA, the antigen was present in homogenized cercariae and schistosomula, but not adult worms, and it was also present in detergent extracts of intact cercariae and schistosomula. Using an antibody-dependent cell adherence assay, anti-K3 serum was found to react with the surface of live cercariae and with the surface of schistosomula recovered from the skin of mice infected up to 48 h previously. Anti-K3 serum also reacted with the surface of S. bovis, S. haematobium and to a lesser extent S. japonicum schistosomula.

Three IgM monoclonal antibodies, M22G11P, M7B7 and M22B3G, which reacted with the surface of Schistosoma mansoni schistosomula in an indirect fluorescent antibody assay, were found to recognize a polysaccharide-containing egg antigen previously designated K3. The monoclonal antibodies and a monospecific rabbit anti-K3 serum also recognized a crossreacting antigen in a crude cercarial antigen preparation. In an eosinophil-mediated schistosomulum killing assay, all three monoclonal antibodies significantly inhibited the level of killing produced by human infection serum. An IgG3 monoclonal antibody, M22C1C, which also recognized the egg antigen K3, did not inhibit eosinophil-mediated killing. However, when lower concentrations of human serum were used in the assay, this monoclonal antibody significantly enhanced the level of killing, despite having no capacity to induce eosinophil-mediated damage in the absence of human infection serum. On the basis of these and other results we suggest the possibility that antibodies to S. mansoni egg antigens which cross-react with the surface of the early post-penetration schistosomulum may influence the effective expression of antibody-dependent, eosinophil-mediated effector mechanisms in human infections.

A total of 129 children were treated for Schistosoma mansoni infections, and followed for intensity of reinfection at 3-monthly intervals over a 21-month period. Blood samples were taken before treatment and at 5 weeks and 6, 12 and 18 months after treatment. This paper presents a statistical analysis of the relationship between various immune responses and subsequent reinfection. Responses analysed were: blood eosinophil levels; IgE antibodies against schistosomulum antigens; IgG antibodies mediating eosinophil-dependent killing of schistosomula; antibodies inhibiting the binding to schistosomulum antigens of two rat monoclonal antibodies that also recognize egg antigens; the levels of anti-adult worm and of anti-egg (total, IgM and IgG) antibodies; and IgM anti-schistosomulum antibodies. Results for each assay were well correlated for each of the five separate blood samples. None of the assays were predictive of resistance to reinfection, but susceptibility to reinfection was strongly correlated with results in the preceding blood samples for total anti-egg antibodies and the inhibition of binding of one of the two monoclonal antibodies. Further analysis also revealed a correlation between reinfection intensities and both IgM anti-schistosomulum antibodies and IgM and IgG anti-egg antibodies. These results are consistent with the hypothesis that early infections elicit the development, in response to egg antigens, of antibodies that block immune mechanisms directed against schistosomula. Blocking antibodies may be IgM, but might also be of an ineffective IgG isotype. The existence of such antibodies in young children would explain the slow development of immunity in the face of a range of detectable, potentially protective immune responses.

Two isolates of Schistosoma mansoni from Puerto Rico and Egypt were examined to determine if there were differences in surface antigens of the schistosomulum and to assess the ability of the two isolates to induce protection against one another in vivo. Immune mouse and human patient antisera recognized the same antigens on the schistosomulum surface of both isolates. However, mice immunized with schistosomula-released products from the Egyptian isolate recognized an additional antigen of Mr 13K on the Egyptian schistosomulum surface which was not present in the Puerto Rican isolate. In quantitative radioimmunoassay, sera from mice vaccinated with irradiated Egyptian cercariae bound more strongly to Egyptian schistosomula than to Puerto Rican parasites. Both isolates cross-protected against each other, but mice were less immune to challenge with Egyptian cercariae after being immunized with Puerto Rican irradiated cercariae. There was no difference in immunity to challenge when Egyptian irradiated cercariae were used to immunize. Although this evidence suggested some heterogeneity within the Egyptian isolate, cloned cercariae of the Egyptian isolate did not vary in their ability to cross-protect against each other. Furthermore, antisera from mice immunized with clones of Egyptian cercariae recognized the same schistosomulum surface antigens. The results reported here indicate that although there were small differences between the two isolates the major surface antigens are conserved.

Immunity to Schistosoma mansoni was investigated in congenitally athymic (Nu/Nu) rats, irradiated rats and in mast cell-depleted rats. Nu/Nu rats failed to develop significant resistance following vaccination with irradiated cercariae, although Nu/Nu recipients of serum from vaccinated Fischer rats (VRS) manifested resistance comparable to heterozygous controls, suggesting that T-cells were required in the induction of resistance but were not involved in the efferent arm of antibody-dependent elimination. Radiosensitive cells (including eosinophils, basophils, neutrophils, lymphocytes and mast cells) were apparently not essential for the antibody-dependent elimination of lung or post-lung stages since irradiated (700–750 rad.) recipients of VRS manifested comparable degrees of resistance to unirradiated controls in spite of a greater than 85% reduction in total blood leucocyte counts after irradiation. Depletion of 99 % of tissue mast cells by treatment of rats with Compound 48/80 had no significant effect on the attrition of a challenge infection in rats rendered immune by vaccination with irradiated cercariae or by transfer of VRS. However, there was a significant increase in worm recovery in unimmunized and mast cell-depleted or irradiated rats, indicating that mast cells and perhaps other radio-isotope sensitive cells may be involved in innate resistance.

Previous workers have reported that significant resistance to Schistosoma mansoni cercarial challenge can be induced in mice by immunization with various Fasciola hepatica antigen preparations. We have attempted to reproduce these observations and have also carried out similar experiments in rats. In our hands, however, CBA mice did not develop either antibodies against the schistosomular surface, or significant resistance to challenge after immunization with heterologous antigens. Similar results were obtained in Fischer rats, but rats did develop significant resistance after immunization with homologous (schistosomular) antigens, even without adjuvant, and the homologous ‘protective’ antigens could be separated by antibody-affinity chromatography using serum from rats vaccinated with irradiated cercariae. Thus, we found no evidence for the existence of shared ‘protective’ antigens between S. mansoni and F. hepatica, and suggest that this is further evidence that where heterologous resistance can be demonstrated between these two parasites, it is probably due to immunologically non-specific mechanisms.

Thirty-two isolates from man in known areas of Gambian trypanosomiasis, in the Sudan, Kenya, Zaire, Nigeria, Ivory Coast, Burkina Faso, Liberia and Senegal, were examined by isoenzyme electrophoresis of 11 enzymes. Comparisons were also made with our previously published results on 23 other stocks of similar origins, which had been examined in the same manner. All those stocks of low initial virulence to laboratory rodents, which thus conform to the accepted view of the behaviour of Trypanosoma brucei gambiense can be identified by characteristic combinations of enzyme patterns, especially certain aminotransferase markers. A limited study of superoxide dismutase polymorphism suggested a further marker of value. The isolates of high initial virulence to rodents, which are thus behaviourally akin to T. b. rhodesiense, did not share these characteristics. We conclude that there exists a homogeneous group of trypanosomes of wide dispersion throughout tropical Africa, characterized by certain isoenzyme combinations and low initial virulence to rodents, which corresponds to the classical concept of T. b. gambiense. The features of limited antigenic repertoire, high resistance to normal human serum and restriction fragment length polymorphisms of the genes for certain variant surface glycoproteins also appear to be characteristic of this group.

A herd of 20 cattle was introduced on a ranch in Kilifi, Coast Province of Kenya, where they were in contact with Glossina austeni for 6 months. In total, 65 trypanosome isolates were made from these animals. Examination of the isolates revealed that 61 were Trypanosoma congolense and 4 were T. theileri. Out of the 61 T. congolense isolates, 55 were successfully passaged and cloned in mice to provide trypanosome populations for further analyses. The stocks and their clones were inoculated into goats on which teneral G. morsitans centralis were later fed in order to provide metacyclics for use in serodeme analysis. Identification of serodemes was carried out by indirect immunofluorescence and neutralization using antimetacyclic hyperimmune sera prepared in mice against metacyclics of cloned trypanosome populations. So far 4 serodemes have been identified in 8 stocks and 7 clones. Each of the 9 stocks contained a mixture of at least 2 of the 4 serodemes identified. Furthermore, stocks isolated sequentially from individual animals contained the same serodemes despite repeated treatment with a curative dose (6 mg/kg body weight) of Berenil between isolations. From the latter finding, it can be inferred that the 4 serodemes were present on the ranch throughout the study period.

Changes in the tegumentary projections of protoscoleces of Echinococcus granulosus, developing within brood capsules, were examined at the ultrastructural level. The original scant covering of spineless, truncated microtriches was supplemented by a population of microvilli covering the entire surface of protoscoleces in the early stages of formation. As differentiation continued, rostellar hooks were formed by enlargement of single large (T 1) microtriches, and normal spined microtriches were produced on the sucker region. These were formed in two ways: by addition of spine material to the truncated microtriches, and de novo followed by up-lifting from the tegumentary surface. A second type of large microthrix (T2) was present in a band immediately below the developing hooks. These microtriches appeared to be formed from a fusion of existing microtriches, but were subsequently lost prior to scolex invagination. The microvilli were also shed from both the sucker and soma regions, to be replaced by microtriches and small blunt elevations respectively.

A small, size selected (0·5–5·0Kbp) genomic DNA library has been constructed in the bacterial plasmid pAT153 using DNA extracted from a human isolate (Kenyan origin) of the hydatid disease organism, Echinococcus granulosus. A panel of taeniid cestode DNAs has been used in conjunction with hybridization and restriction-enzyme analysis to identify in the library two recombinant plasmids with Echinococcus-specific inserts and a single recombinant plasmid (coded pEG18) with a DNA fragment unique for E. granulosus. These, and other recombinant plasmids with E. granulosus DNA inserts, have been used in restriction endonuclease, Southern transfer and hybridization analysis to independently and repro-ducibly discriminate between the UK horse and sheep strains of E. granulosus; these probes may well prove of value for characterizing isolates from other endemic areas. The feasibility of using a cloned DNA fragment as the basis of a simple field test for distinguishing eggs of E. granulosus from those of other taeniid cestodes is discussed. The recombinant insert (approximately 2·3 Kbp in size) of pEG18 has a low copy number - estimated at approximately 26 - and it may not be sufficiently sensitive for practical use as a DNA probe in the identification of small numbers of E. granulosus eggs by molecular hybridization.

Lambs aged 48–150 days which had been injected with extracts enriched in a functional antigen (contortin) obtained from adult Haemonchus contortus developed specific circulating antibodies and were less susceptible to haemonchosis when challenged 1 month later with a single dose of 20000–25000 infective 3rd-stage larvae. Sera from 18 out of a total of 19 lambs injected with the extract contained precipitating antibodies to 2–5 components of the extract. None of these lambs died. The one extract-injected lamb which did not develop antibodies and 9 of the 13 lambs used as controls died of haemonchosis. The average weight of worms recovered was 1·45 g from the immune lambs and 5·72 g from the non-immune lambs.

The Jamaican iguanid lizard Anolis lineatopus is the host of 4 species of helminth parasite: an acanthocephalan (Centrorhynchus spinosus), a digenean (Mesocoelium danforthi), and 2 nematodes (Cyrtosumum scelopori and Thelandros (?cubensis). Prevalance of C. spinosus was unrelated to host age but was considerably higher in males (43%) than in females (10%). The intensity increased with host age: (range: 1–23, median: 3). Prevalence of C. scelopori increased with host age in both sexes: juveniles (<3 months) were uninfected while infection occurred in almost all lizards older than 9 months. Intensity was higher in males (range: 1–403, median: 158) than in females (range: 1–297, median: 86). The infection patterns of C. spinosus and C. scelopori differed significantly from each other and were not consistent with a simplistic direct relationship between time of exposure and infection prevalence. Acanthocephalan and digenean infections occurred at low prevalence and intensity although prevalence in older, larger lizards tended to be higher than in younger, smaller ones.

The caecal mucosa of wild young and adult grouse infected naturally with Trichostrongylus tenuis was examined by means of scanning electron microscopy and compared with adult grouse which had been treated with an anthelmintic. The caecal mucosa of young red grouse with low worm burdens possessed longitudinal plicae and exhibited little damage. The caeca from adult grouse, most of which carried high worm burdens, showed a localized depression of plicae and atrophy and cell disruption in areas of nematode aggregation. Caeca from adult birds treated with an anthelmintic showed a similar caecal structure to lightly infected caeca from young birds. It is concluded that the normal functioning of the caeca is probably affected by heavy infections of T. tenuis.