The pattern of recovery of schistosomula from the lungs of mice, hamsters and rats is described following normal percutaneous infection via the abdominal skin. Peak numbers were found in hamsters and rats on day 5 and in mice on day 6 post-exposure. Schistosomula were recovered from the lungs up to approximately day 20 post-exposure in all 3 species. None were found in pleural washings and only 2 were recovered following mincing and incubation of host diaphragms. The pattern of recovery of schistosomula from the lungs of mice was also described following the injection of a pulse of parasites into the tail vein. Approximately 54% of these injected schistosomula eventually reached maturity in the hepatic portal system. The ability of schistosomula to migrate and mature following their injection into unusual locations was tested. Small numbers reached maturity in the hepatic portal system following injection into the pleural cavity or subcutaneously, more when injected intra-peritoneally and largest numbers following intravenous injection. Small numbers of schistosomula were recovered by mincing and incubation of systemic organs such as the kidney and spleen. Schistosomula injected into the left ventricle of hamsters were able to migrate via the systemic organs to the lungs and 47% eventually matured in the hepatic portal system. The pattern of schistosomulum accumulation in the hepatic portal system of mice, hamsters and rats is described. Following normal percutaneous infection, schistosomula were first detected in this site from day 6 to day 9 depending on the host species. In mice, migration was complete around day 20 but continued in hamsters up to day 40 post-exposure. Following injection of schistosomula into the tail veins of mice, the first arrivals in the hepatic portal system were detected 12 h later and were found to lose their ability to migrate shortly after arrival. However, a proportion of lung schistosomula injected directly into the hepatic portal system were able to traverse hepatic sinusoids and reach the lungs. It was concluded that the route of migration of schistosomula from lungs to liver was entirely intravascular, with potentially several passages round the pulmonary-systemic circulation, before chance entry into arteries leading to the hepatic portal system. The proportion of schistosomula exiting from the pulmonary-systemic circuit was estimated as 0·14/day. A computer simulation produced values of 11 h and 5 h for the duration of migration through pulmonary and systemic capillary beds. The time for 1 complete circuit would thus be 16 h and the proportion of schistosomula exiting per circuit would be 0·095. This approximates to the proportion of cardiac output going to splanchnic organs in the resting rat (0·128). The narrowness of hepatic sinusoids may be one factor contributing to the sequestration of schistosomula in the hepatic portal system.

Homologous and heterologous resistance in Echinostoma revolutum infections was studied in mice. A high level of homologous resistance was demonstrated in mice harbouring a 13-day-old primary E. revolutum infection with 9–10 and 11–15 worms, corresponding to a 70·0 and 66·7% reduction in the size of the established worm burden as compared with that of the challenge control group. A 14- and 20-day-old primary infection with 3–4 worms induced a level of resistance of 61·7 and 81·8% respectively, while higher worm levels of 9–10 and 11–15 induced almost complete resistance corresponding to a 95·1–100% reduction in the size of the established worm burden. Complete resistance was also demonstrated in mice challenged 8 days after elimination of a 20-day-old primary infection with 11–15 worms by anthelmintic treatment. A primary 43-day-old Schistosoma mansoni infection induced a 73·1% reduction in the size of the established E. revolutum challenge infection while infections of an age of 79 and 99 days conferred complete resistance to heterologous challenge with E. revolutum. Primary pre-patent S. mansoni infections and a patent S. bovis infection of an age of 56 days did not induce any resistance to challenge with E. revolutum. A primary 14- and 21-day-old infection of E. revolutum did not stimulate any significant level of resistance to heterologous challenge with S. mansoni.

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.

Intraperitoneal injection of cercariae into pristane (2, 6, 10, 14 tetramethyl pentadecane)-primed Balb/c mice led to greatly diminished numbers of portal and peritoneal worms compared with untreated mice. Schistosomula taken from the peritoneal cavity of pristane-primed mice carried globules of pristane on their surfaces, were contracted and were permeable to Trypan blue. Pristane globules bound also to adult worms in vitro and in vivo causing rapid damage to the surface membrane. Hydrophobic compounds other than hydrocarbons either bound without causing gross damage, or did not bind to the adult worms. 51Cr release studies showed that pristane had no effect on the permeability of human erythrocytes, while causing significant release from both schistosomula and adult worms. The binding of hydrocarbon globules to a variety of other parasites did not occur. The binding of n-[1-14C]hexadecane to adult Schistosoma mansoni was significantly decreased by extraction of the parasite with organic solvents or treatment with staphylococcal δ toxin, which interacts with phospholipids in the membrane. Possible mechanisms of damage of the parasite by the hydrocarbons are discussed.

A technique is described for the isolation of a pure suspension of second-generation schizonts of Eimeria necatrix and E. tenella. Most of the schizonts appeared viable as judged by their ability to exclude Trypan blue and the use of this criterion is examined. They were able to metabolize radio-isotope labelled glucose and accumulate thiamine against a concentration gradient. A formula allowing detailed consideration of the latter process is introduced. Crude extracts of these schizonts exhibited serine hydroxymethyl transferase activity which is significantly different from that of the host tissue in terms of optimal pH (8·3 for the parasite and 7·8 for the avian intestinal cells) and Michaelis constant (approximately 2·6 × 10−3m and 0·6 × 10−3m respectively). Differences in response to potential inhibitors are also shown.

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.

Incorporation of labelled amino acid into tegumental proteins and acquisition of protection by schistosomula against antibody-mediated killing in vitro were simultaneously stimulated by serum factors and inhibited by puromycin. Comparison of polyacrylamide gel electrophoresis patterns with fluorographic autoradiography indicates that the majority of proteins in the parasite tegument were labelled with the isotope after incubation for 3 h. No new, clearly defined band was observed in the autoradiography pattern. During this period a decreasing susceptibility of the schistosomula to antibody plus complement was observed. Quantitative fluorescence assay shows that schistosomula insensitive to antibody plus complement were still able to bind the same amount of antibody as the unprotected parasites. Pre-culture of schistosomula in the presence of inactivated normal rabbit serum also decreased the susceptibility of the parasites to the in vivo killing mechanism.

Amastigotes from Leishmania braziliensis and L. mexicana were isolated from cutaneous lesions in infected animals using the plant lectin Concanavalin A as a specific agglutination agent. Amastigotes were collected in preparations of up to 95% purity as determined by cell count. The parasites obtained by this method showed no apparent loss of viability as measured by growth rates and DNA replication, or pathogenicity as measured by routine passages in hamsters or mice.

The uptake of thiamine by isolated second-generation schizonts of Eimeria tenella and by host intestinal cells was found to consist of two components. One was passive and the other apparently an active process. The kinetic constants of the latter were compared in host (Km = 0·36 μm) and parasite (Km = 0·07 μm) and found to be significantly different. Both systems were competitively inhibited by amprolium but showed different affinities for the drug. (Host Ki = 326 μm; parasite Ki = 7·6 μm). Further differences were observed in schizonts of a drug-resistant line of E. tenella. These findings are discussed in terms of the inhibition of thiamine uptake being the basis of the anticoccidial activity of amprolium.

The influence of Meloidogyne javanica on cytokinins in the host Lycopersicon esculentum has been studied at different stages of the nematode's life-cycle. Marked differences were detected in cytokinin content of root homogenates between infected and control plants, particularly at the 3rd (32 day), 4th (39 day) and 5th (55 day) harvests. Most of the cytokinin detected appeared to be associated with root homogenates in which the nematode was in the rapid post-moult growth stage. The influence of these nematodes on cytokinins in the host's xylem exudate was not nearly so pronounced. The freshly hatched 2nd-stage infective larvae of M. javanica were themselves capable of exuding cytokinin-like substances.

Extracts of rhesus monkey erythrocytes infected with Plasmodium knowlesi were fractionated by polyacrylamide gel electrophoresis (PAGE) and several zones of endopeptidase activity were demonstrated by an imprint-digest method. The enzymes were active only under acid conditions; activity was detected at pH 3·2 but not between pH 6·4 and 8·9 using haemoglobin, albumin or erythrocyte lysate as the substrate. Optimized PAGE conditions separated highly active parasite enzymes with Rf values of 73, 63 and 53 (± 7%), as well as a red cell endopeptidase, Rf44. Of two other minor bands of activity, one was associated with platelets.

Glycoproteins of Trypanosoma congolense have been detected on SDS-polyacrylamide gels using the Concanavalin A peroxidase technique. Using [35S]diazoniobenzenesulphonate as a marker for cell surface proteins it was possible to distinguish between internal glycoproteins and the surface coat proteins. On SDS-polyacrylamide gels Con A reacted with the surface coat proteins. Results obtained from Con A-induced agglutination of living trypanosomes indicated that sugars of the surface coat proteins were accessible to Con A. This was reinforced by the cytochemical visualization of Con A binding to the trypanosome surface. The results suggested that the surface coat protein contained α-linked d-mannosyl, d-glucosyl, or N-acetyl-d-glucosaminoyl residues, which are exposed exteriorly on the surface coat.

A procedure is described for the isolation of sub-cellular fractions from bloodstream forms of Trypanosoma brucei. The method leaves intact most of the nuclei, mitochondria and microbodies. All the fractions have been chemically characterized and tested for 10 enzymatic markers. About 5% of total cell protein was isolated as a microsomal fraction containing mostly plasma membranes and endoplasmic reticulum vesicles. Plasma membranes were purified by high-speed centrifugation on magnesium-containing Dextran, and on linear sucrose-density gradients. The yield of membranes was approximately 0·3% of the total cell protein. The purified material had a sucrose density of 1·14 g/cm3 and consisted of smooth vesicles. Specific activity of the membrane markers Na+, K+, ouabain-sensitive ATPase and adenylate cyclase were 26-and 20-fold higher, respectively, than in total cells. Neither DNA nor RNA was detected. The sum of the cholesterol and phospholipid content was 0·99 mg/mg protein. The cholesterol/phospholipid molar ratio was 1 : 2.

A model has been developed to simulate the surface of an antibody-coated schistosomulum. It consists of a layer of agar, containing antigen (tetanus toxoid) and a chemotactic factor (ECF). Some layers were coated with human anti-tetanus immunoglobulin. The mode of adherence of human eosinophils and neutrophils to these agar layers and the subsequent degranulation of the cells exactly paralleled the interaction of these cell types with antibody-coated schistosomula of Schistosoma mansoni. In particular, eosinophils made much more intimate contact than did neutrophils, and lysosomal enzymes were secreted extracellularly by direct fusion of granules with the plasma membrane of the cell. Biochemical evidence was also obtained for the secretion of enzymes during degranulation and the rate of enzyme release was found to be enhanced in the presence of specific antibody. This model, non-phagocytosable surface has the potential to provide basic information on the mode of action of effector cells in cell-mediated cytotoxic reactions against a wide range of parasites by incorporation of different factors into the agar layers.

Cyclical transmission of different variable antigen types of Trypanosoma congolense STIB 228 resulted in the development of metacyclic trypanosome populations which were similar in their variable antigen composition as judged by immunofluorescence and neutralization assays. The variable antigen types present in the ingested bloodstream populations were not found in the metacyclic populations. The bloodstream populations which were obtained from cyclically infected, irradiated (900 rad.) mice contained variable antigen types which were not present in the corresponding metacyclic populations. When derivatives of 2 other stocks of T. congolense, isolated in a different area of Tanzania, underwent cyclical development in the tsetse fly, the metacyclic populations of each stock also had a characteristic variable antigen composition. The metacyclic populations of the 3 stocks were, however, completely dissimilar in variable antigen composition. Simultaneous infection of tsetse flies with a mixture of different stocks resulted in the concurrent production of metacyclic trypanosomes which contained the characteristic variable antigen types of each stock. The effect of cyclical transmission on the process of antigenic variation in T. congolense infections is therefore similar to that in T. brucei infections.

A set of n-(9-anthroyloxy) fatty acids (n = 2, 6, 9, 12, 16) have been used as fluorescent probes to examine the lipid environment at different depths in the outer membrane of normal mouse erythrocytes and red blood cells from Plasmodium berghei-infected blood. Fluorescent polarization experiments with normal mouse erythrocytes have demonstrated a typical gradient in microviscosity from the surface to the centre of the bilayer as a consequence of the motional properties of the C-atoms of the phospholipid acyl chains. The fluorescent probes rotate faster in the membrane of purified pluriparasitized cells (> 90% purity) than with the remaining fraction of red blood cells from infected blood (20–40% immature, infected red cells, and uninfected red cells), or normal mouse erythrocytes. This increase in fluidity with heavily infected cells occurs predominantly at the centre of the lipid bilayer, rather than at the membrane surface. A comparison of the polarization values of intact and lysed infected cells indicates that the fluorescent fatty acids preferentially label the plasma membrane rather than the internal membranes of infected cells. The results suggest that P. berghei infection causes a change in the composition and/or organization of the outer membrane of pluriparasitized cells which produces a decrease in membrane microviscosity.

African trypanosomes can undergo antigenic variation and evade the host immune response. Whether the antigenic variants arise in an ordered sequence or randomly has been in dispute but has not been statistically tested. The coefficient of concordance (W), a statistic designed to detect similarities between sequences of objects, was applied to the literature data. The tendency towards a reproducible order of variants was strong, although in several of the studies the number of experimental animals was so low that no conclusions could be drawn. A computer model was used to determine whether this degree of order could arise with random generation of variants followed by selection. The model simulated a trypanosome clone with 90 possible variants, widely differing variant-specific growth rates, random variant origin and variant eradication by an anamnestic host immune response. Parameters varied were maximum parasitaemia, growth rate differential between ‘fast’ and ‘slow’ variants, and parasitologist ability to detect minor variants. Random generation and selection by growth rate alone could not produce the degree of variant orderliness reported in the literature. However, experiments with larger numbers of host animals and direct investigation of variant growth rates and competitive interactions are necessary before the random generation-selection hypothesis can be proven or disproven.

Groups of mice were exposed to multiple bites by tsetse flies (Glossina morsitans morsitans) infected with a clone of Trypanosoma congolense spread over a period of 8 days. The mice were subsequently treated with Berenil 10 days after the first fly bite as were uninfected control mice. The group of mice which received 12–15 infectious fly bites on two occasions, 21 days apart, were subsequently resistant to infection when re-challenged by flies infected with the same clone of T. congolense. These mice were also immune to challenge by flies infected with a different bloodstream variable antigen type derived from this same stock. The immunity was stock-specific and directed against the metacyclic forms of the parasite, but was short-lived.

Plasmodium knowlesi merozoites were prepared by the polycarbonate sieving method of Dennis, Mitchell, Butcher & Cohen (1975). Merozoite function was assayed by their attachment to and invasion of rhesus erythrocytes at 37 °C. The early merozoites from the culture chamber were the most invasive, although maximum numbers of merozoites appeared later. Merozoites were most stable when incubated at room temperature (23 °C). At 37 and 0 °C invasiveness rapidly declined to zero. Attachment was rapidly lost at 37 °C but was retained at 0 °C. Attachment was unchanged in the pH range 6·8–7·9 but invasion was reduced at pH 7·9. The presence of l-fucose, d-galactose, d-glucose, d-mannose, N-acetyl-d-galactosamine or N-acetyl-D-glucosamine did not reduce invasion. Attachment and invasion were greatly reduced or abolished by the presence of 2·5 mm EDTA or EGTA, by lactoper-oxidase-catalysed iodination of the merozoites, or by treatment of the merozoites with trypsin at a concentration of 1 μg/ml or greater for 10 min at 23 °C.

Eighteen clones of variable antigen type 052 of Trypanosoma brucei stock S. 427 were derived and maintained as animal-infective bloodstream forms in vitro for up to 60 days of cultivation. The antigenic composition of such clones was monitored weekly by immunofluorescent analysis of viable trypanosomes, using antisera raised to isolated variant-specific surface glycoproteins of both 052 and a variable antigen type (221) which consistently appeared in the first relapse population of type 052 in vitro. The appearance of new variants was detected in 9 of the 18 clones 18–46 days following initiation of the clone and variable antigen type 221 was found in all 9 clones. On one or more occasions in 8 of such clones, viable trypanosomes were found which did not react with either antiserum but were mouse-infective on the 4 occasions tested and probably represent other variable antigen types. The process of antigen, variation in vitro appears to resemble the process in vivo except that new variant types are detected earlier in vivo. This possibly results from different growth rates of the trypanosomes in vivo and in vitro, together with the fact that elimination of the initial variant population by the host's immune response facilitates the detection of newly arising variable antigen types in vivo.