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.

A panel of immunologically and biochemically defined monoclonal antibody probes has been used in conjunction with immunocytochemical techniques to localize target antigens in sections of different life-cycle stages of Trichinella spiralis. Monoclonals that immunoprecipitate surface components from adult worms, show reactivity with the surface but not with internal tissues of sectioned parasites. Reagents that immunoprecipitate radio-isotope labelled stage-specific surface components of muscle-stage larvae, however, react with the stichosome and gut lining of sectioned larvae, as well as with the surface. Monoclonal antibody probes that do not stain the surfaces of live, intact muscle larvae in immunofluorescence assays, but which immunoprecipitate solubilized surface glycoproteins, also show reactivity with cuticular and stichosomal antigens of sectioned larvae. The more powerful resolution provided by electron microscopy has localized the surface antigens to the epicuticle and the intestinal antigens to the brush-border microvilli. Of particular interest was the finding that antigens of muscle-stage larvae, known to confer protection upon recipient mice, also exist in the stichosome of adult parasites. This observation may shed some light on the fact that mice immunized with antigens from muscle-stage larvae show, in addition to reduced muscle larva burden, accelerated expulsion of adult worms. The implications of these data for stage specificity of immune responses to trichinosis are discussed.

Throughout the world, infection with parasites is extremely common. Prevalence is highest in the warm countries of the tropics and subtropics, but infection occurs in all climatic zones. The figures usually quoted for the prevalence of specific parasites attain values akin to those used to describe astronomical distances. The World Health Organization (1985) estimated that some 2582 million of the earth's inhabitants live in areas where they risk contracting malaria and that many – in Africa south of the Sahara alone probably 200 million – remain chronically and persistently infected. Estimates for other parasites are similarly impressive; 600 million at risk of infection with schistosomiasis, with some 200 million actually infected; 900 million exposed to lymphatic filarial parasites, with some 90 million currently infected; for onchocerciasis, probably some 40 million cases spread throughout Africa, Central and South America and the Eastern mediterranean. Yet, impressive as these estimates may be, they are dwarfed by those for some intestinal helminths. Recent assessments suggest that 1 person in 4 of the world's population carries Ascaris lumbricoides (Schultz, 1982), while the same proportion is believed to be infected with one or other of the hookworms (Gilman, 1982).

The influence of subcutaneous infection with protoscoleces of Echinococcus granulosus on the distribution of lymphocyte subsets in draining lymph nodes has been evaluated by immunocytochemical labelling of lymphocyte surface antigens. These studies reveal marked expansion of paracortical Thy-1 +, Lyt-1 + cells and moderate proliferation of surface immunoglobulin-bearing B-cells immediately after infection. The Lyt-1 +:Lyt-2 + ratio decreases rapidly during the first 21 days post-infection and remains below unity until at least 12 weeks post-infection due to severe depletion of Lyt-1 + cells in draining lymph nodes and a significant increase in the percentage of Lyt-2 + cells. The potential role of these Lyt-1 ∓, 2 + (putative T-suppressor) cells in regulation of the anti-parasite immune response and mediation of generalized immunosuppression is discussed in the light of evidence of inhibition of anti-parasite immunity in infected mice.

Experiments to investigate the metabolism of glycine, L-glutamic acid and L-aspartic acid by Moniliformis moniliformis were carried out by incubating adult worms aerobically for 3 h at 37°C in Tyrode's solution containing either [U-14C]glycine, L-[U-14C]glutamic acid, L-[U-14C]aspartic acid or L-[4-14C]aspartic acid. Much of the glycine and glutamic acid was absorbed by the worms, but little of either was metabolized. Aspartic acid was readily taken up and metabolized. After incubating with L[U-14C]aspartic acid, most radioactivity was found in ethanol and a volatile compound, presumed to be carbon dioxide, with smaller amounts in lactate, alanine, acetate, malate, glucose and succinate. After incubating with L-[4-14C]aspartic acid, most radioactivity was found in lactate and the presumed CO2 with small amounts in alanine, malate and succinate. No radioactivity was found associated with ethanol or acetate. Possible metabolic pathways and suggestions for a relationship between the metabolism of aspartate with that of alanine and serine in this parasite are discussed.

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.

Anti-thymocyte serum (ATS) was administered to CD-1 mice infected with 100 Trichinella spiralis larvae and its effects on intestinal lysophospholipase (EC 3.1.1.5) activity, peripheral blood lymphocytes and bone marrow, peripheral blood and intestinal eosinophilia were assayed in the same experimental animal. The ATS caused a significant suppression of both the tissue lysophospholipase response and eosinophilia, in all three compartments, when compared to the values found in infected mice that were either treated with normal rabbit serum (NRS) or untreated. The suppressed eosinophil response and reduced lysophospholipase activity demonstrated a close temporal relation throughout the experiment. These findings support the hypothesis that helminth parasite-induced eosinophilia is the cause of increased lysophospholipase activity present in parasitized tissue and that the responses are thymus cell dependent.

Good wine needs no bush, and good papers need no third-party summary. The papers presented today at this symposium on the pathophysiology of parasitic disease have been splendid, and I shall not be so foolish as to attempt the impossible task of summarizing them. This stage in the meeting does, however, provide an opportunity to look forward in the light of what has been presented today, and there are some points that are of particular interest and which will bear repetition or comment in the closing minutes of this symposium.

Phosphorylcholine (PC), an immunodominant component of the cell wall of certain bacteria, fungi and nematodes, is known to induce low anti-PC antibody levels during natural infection by Trichinella spiralis. This article reports a study in which spleen cells from BCF1 mice infected with Trichinella sp. larvae were found to produce large numbers of direct haemolytic plaques in response to PC conjugated to sheep red blood cells (SRBC) after muscle-encysted larvae had been killed by treatment with mebendazole. Inhibition of the response by PC-chloride, immunodiffusion and immunoelectrophoretic studies with the anti-PC IgA (TEPC-15) and anti-idiotype T15 serum assays showed the plaque-forming cell (PFC) response to be specific for PC. The absence of haemolytic plaques when unconjugated SRBC or TNP-SRBC were used as indicator cells ruled out involvement of a polyclonal response. Greatest anti-PC PFC response was found to be associated with a microsomal fraction designated FCpl, a particulate fraction behaving as a thymus-dependent antigen. The FCpl fractions from all four strains of Trichinella employed induced anti-PC PFC responses when injected into mice. These results suggest that FCpl is a suitable antigen for use in detailed studies of immune responses to Trichinella and related parasites.

The general structure and aspects of the development of the ovaries of the palaeacanthocephalan Corynosoma semerme were studied by transmission electron microscopy using worms varying in age from 18 to 90 h obtained from experimental primary infections in hydrocortisone-treated rats instead of seals, which serve as the natural definitive hosts. The observations can be interpreted to show that the immature ovaries become transformed relatively rapidly from cellular spheres to the more complex mature ovaries consisting of the supporting and oogonial syncytia and the germ-line cells. The supporting syncytium developed before the oogonial syncytium. The cytological appearance of ovaries from worms from rats was found to be similar to that of ovaries from mature worms of unknown age from naturally infected seals. Ecological evidence is considered to suggest that the rapid maturation of C. semerme in rats may also occur when the parasite becomes established in seals.

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.

The electrophysiological properties of the bag region of the somatic muscle cells of Ascaris suum and Ascaridia galli were studied using intracellular techniques. For Ascaris muscle cells, the mean resting membrane potentials at 20 and 37°C were −29·9 and −33·8 mV respectively, and the average input conductance was 2·12 μS. For the muscle cells of A. galli similar values were obtained. For example, the mean input conductance of these cells was 2·84 μS at 20°C. Healthy Ascaris muscle cells at near physiological temperatures show both spontaneous depolarizing and hyperpolarizing activity and, in cells close to the nerve cords, rhythmic large amplitude (approximately 30 mV) action potentials are observed. Such action potentials, which are very sensitive to temperature variations, originate in the muscle cells. In contrast the muscle cells of Ascaridia are quiescent. The rhythmic action potentials of Ascaris are resistant to tetrodotoxin (TTX) (≤ 10−6 M), verapamil (10−4 M) and cinnarizine (10−4 M), but are blocked irreversibly by 22, 23 dihydroavermectin B1a (10−7 to 5 × 10−6 M). GABA, and the GABAA receptor agonists, muscimol and isoguvacine, hyperpolarize and increase the input conductance of both Ascaris and Ascaridia muscle cells. The antagonists+bicuculline and picrotoxin were not effective in modulating the spontaneous hyper polarizations of Ascaris muscle cells, and picrotoxin (10−4 M) was not effective in altering the response to GABA (5 × 10−6 M). The significance of the results is discussed briefly.

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.

The effects of the phenothiazine, Stelazine, on Hymenolepis diminuta were investigated. The cestode was incubated for 10 min at 37 °C with 1 mM trifluoperazine, in the presence and absence of Ca2+. Assay of brush border enzymes showed that drug treatment lowered the activities of alkaline phosphatase, Ca2+-ATP'ase, 5′-nucleotidase and type 1 phosphodiesterase. This occurred in parallel with a significant reduction in tegumental protein. Under these conditions gross changes in ultrastructural appearance and cellular organization were observed. There was a lack of ordered microtriches and the distal cytoplasm was absent. Glycogen granules were scattered throughout the cytoplasm within the subtegumental layer. The connective tissue also appeared to be in some disarray. The effects of Stelazine appeared to be dependent on time and were significantly increased when Ca2+ was included in the incubation medium. Incubation with the less hydrophobic phenothiazine trifluoperazine sulphoxide had minimal effect on the integrity of the cestode. The results reported here support the premise that certain phenothiazines may be considered as potential cestocidal agents.

Rats harbouring 35-day-old primary infections of Moniliformis moniliformis and Hymenolepis diminuta were inoculated with equal doses of Nippostrongylus brasiliensis and were autopsied 10 days later. Significant reductions were found in the dry weight of Moniliformis and Hymenolepis and in the numbers of Nippostrongylus recovered compared with single infections. Similarly, in infections with two parasites, the numbers of Nippostrongylus were reduced when concurrent with Moniliformis and the dry weights of Moniliformis and Hymenolepis decreased in the presence of each other. A comparison of the concurrent infections themselves revealed that Moniliformis weighed significantly less in the presence of Hymenolepis than in the three parasite infections. Parasite interactions and their possible mechanism are discussed and comparisons are made with the relevant single infection for each parasite.

The cestodes Echinococcus granulosus, Taenia multiceps, T. pisiformis, T. hydatigena, Hymenolepis diminuta, Moniezia expansa and Anoplocephala perfoliata all produced substances that stimulated thymidine incorporation by whole blood lymphocyte cultures. This mitogenic activity was demonstrated in metacestode cyst fluids, live protoscoleces and scoleces, parasite culture supernatants, and extracts of adult parasites. T. multiceps metacestode cyst fluid mitogen adhered to, but would not pass through, cellulose dialysis tubing. This adherence was reduced or prevented by D-glucose and by proteins. The mitogen was weakly anionic. With Sephadex G75 gel filtration, its elution volume was greatly decreased when the elution buffer contained 0·5 M D-glucose. Mitogenic activity was protease resistant and could be separated by gel filtration from all the cyst fluid proteins.

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.