The nature of parasitism at the population level is defined in terms of the parasite's influence on the natural intrinsic growth rate of its host population. It is suggested that the influence on this rate is related to the average parasite burden/host and hence to the statistical distribution of parasites within the host population.

Theoretical models of host–parasite associations are used to assess the regulatory influence of parasitic species on host population growth. Model predictions suggest that three specific groups of population processes are of particular importance: over-dispersion of parasite numbers/host, density dependence in parasite mortality or reproduction and parasite-induced host mortality that increases faster than linearly with the parasite burden. Other population mechanisms are shown to have a destabilizing influence, namely: parasite-induced reduction in host reproductive potential, direct parasite reproduction within the host and time delays in the development of transmission stages of the parasite.

These regulatory and destabilizing processes are shown to be commonly observed features of natural host-parasite associations. It is argued that interactions in the real world are characterized by a degree of tension between these regulatory and destabilizing forces and that population rate parameter values in parasite life-cycles are very far from being a haphazard selection of all numerically possible values. It is suggested that evolutionary pressures in observed associations will tend to counteract a strong destabilizing force by an equally strong regulatory influence. Empirical evidence is shown to support this suggestion in, for example, associations between larval digeneans and molluscan hosts (parasite-induced reduction in host reproductive potential counteracted by tight density-dependent constraints on parasite population growth), and interactions between protozoan parasites and mammalian hosts (direct parasite reproduction counteracted by a well-developed immunological response by the host).

The type of laboratory and field data required to improve our understanding of the dynamical properties of host–parasite population associations is discussed and it is suggested that quantitative measurement of rates of parasite-induced host mortality, degrees of over-dispersion, transmission rates and reproductive and mortality rates of both host and parasite would provide an important first step. The value of laboratory work in this area is demonstrated by reference to studies which highlight the regulatory influence of parasitic species on host population growth.

Eimeria grenieri was isolated from intensively reared guinea fowl in Britain. The oocysts had average dimensions of 21·09 × 15·48 μm and a small micropyle. Three generations of schizonts were found in the small intestine. The first mature schizonts were found in the upper intestine 30 h after inoculation. Second-generation schizonts were seen 48–80 h and third-generation schizonts were present 80–96 h after inoculation. These schizonts were found in the middle and lower small intestine. It was confirmed that gametogony of this species occurs only in the caeca. The pre-patent time was 112 h and the reproductive index from a dose of 6 × 102 oocysts was 8 × 104. Doses of between 1 × 104 and 1 × 106 caused severe depression of body weight gain. Immunity to re-infection developed rapidly. Sulphaquinoxaline at a dose of 0·04% in the drinking water given after inoculation was effective in reducing the pathogenic effects of the disease. Robenidine given in the food at a concentration of 16·5 ppm (50% of the recommended concentration for chickens) was effective in preventing disease caused by E. grenieri.

The endogenous life-cycle of E. grenieri was completed in the chorioallantoic membrane of developing chicken embryos.

Electrophoretic variation of the enzymes glucose phosphate isomerase, 6-phosphogluconate dehydrogenase, lactate dehydrogenase and glutamate dehydrogenase (NADP-dependent) has been studied in the African murine malaria parasites Plasmodium berghei, P. yoelii, P. vinckei and P. chabaudi and their subspecies. Horizontal starch gel electrophoresis was used throughout. The number of isolates examined in each subspecies varied from 1 (P. y. nigeriensis) to 24 (P. c. chabaudi). Extensive enzyme variation was found among isolates of most of the subspecies from which more than two such isolates were available for study. It is clear that the phenomenon of enzyme polymorphism is of common occurrence among malaria parasites. With the exception of P. berghei and P. yoelii, of which all isolates share an identical electrophoretic form of lactate dehydrogenase, no enzyme forms are shared between any of the 4 species of murine plasmodia. By contrast, within each species common enzyme forms are shared among each of the subspecies. The subspecies are, nevertheless, distinguished from each other by the electrophoretic forms of at least one enzyme.

The distribution and reassortment of enzyme variation among isolates of a single subspecies is in accordance with the concept of malaria parasites as sexually reproducing organisms. The study of variation among parasites present in individual wild-caught rodent hosts demonstrates that natural malarial infections usually comprise genetically heterogeneous populations of parasites. Nevertheless, the number of genetically distinct types of parasite of any one species present in a single infected host appears to be small. Generally not more than 2 or 3 clones of parasite of distinct genetic constitution are present in a single infected animal.

Vitamin K (as menaphthone sodium bisulphite) added to a deficient diet reduced mortality due to Eimeria tenella or E. necatrix, had a slight effect on haematocrit, but had no obvious effect on weight gain or faecal blood; 0·1 ppm gave a maximal response. The effect of vitamin K on mortality was not absolute; the magnitude of the effect depended on the size of the challenge dose of oocysts. Likewise, the response of an infection to anticoccidial drugs, particularly monensin, depended on the severity of challenge. The effect of adding vitamin K in the presence of drug was to effectively reduce the coccidial challenge; no other interaction of vitamin K and drug has been found. No effects with vitamin K deficiency or supplementation were seen in cases of infections with E. acervulina, E. brunetti or E. maxima. The use of a deficient diet for experimental work is therefore quite justified – particularly as it results in a 4-fold saving of oocysts for inoculation purposes in the case of the haemorrhagic species.

Isolation of blood and intracellular forms of Trypanosoma cruzi was made mainly from rats (90–110 g) which had received 580 rad of whole-body γ-irradiation not more than 24 h before subcutaneous inoculation with 107 trypomastigotes of the Sonya strain of T. cruzi. Unirradiated chinchillas (250–350 g) were, however, used for some experiments. Blood forms were isolated using a technique involving differential centrifugation to remove most of the erythrocytes and DEAE–cellulose chromatography to remove the remaining blood cells. Overall recoveries were usually in the range 30–70%. Parasites were mainly (approximately 98%) broad forms and were motile, metabolically active (as judged by respiratory and radio-tracer incorporation studies) and had lost none of their infectivity for mice. Intracellular forms were isolated from hind-limb muscle tissue. This was disrupted in an MSE tissue homogenizer and the homogenate incubated with DNase, collagenase and trypsin. Parasites, contaminated only by a few blood cells, were then obtained by differential centrifugation. For purer preparations, a terminal sucrose gradient step was used. Recoveries ranged between 40 and 70%. About 1–3% of the parasites isolated were epimastigotes and trypomastigotes; the remainder are probably best collectively termed ‘amastigotes’, though they were pointed and most had a short, free flagellum. They were undamaged as judged by light and electron microscopy and metabolically active as judged by respiratory and radio-tracer incorporation studies. However, the infectivity for mice of both these purified preparations and the initial cell homogenates could be accounted for by the epimastigotes and trypomastigotes present in them. Preliminary biochemical studies with isolated parasites have shown that blood, intracellular and culture forms of T. cruzi have a respiratory system which is in part sensitive to CN- and that all forms synthesize nucleic acids and proteins when incubated in vitro. There appears, however, to be a lack of DNA synthesis in blood stages, and thus it is not surprising that these forms do not divide.

The succinate dehydrogenase of adult Fasciola hepatica was found to exist in active and inactive forms. The enzyme was inactivated by 1 μM oxaloacetate and activated by incubation with compounds which bind to the active site (succinate, fumarate, malonate) or by incubation with anions and certain nucleotides. The activation of the enzyme by succinate followed first-order kinetics. The extent of activation of F. hepatica succinate dehydrogenase depended on the nature and concentration of the activator and on the pH. The rate of activation of the enzyme depended on the temperature. In contrast, the fumarate reductase activity of F. hepatica was not activated by incubation with substrate or anions and was not inhibited by oxaloacetate (100 μM). The significance of these results in the regulation of the tricarboxylic acid cycle in parasitic helminths is discussed.

Sheep infected with adult Fasciola hepatica were drenched with rafoxanide. At 4, 8, 16 and 24 h after drenching the sheep were killed and the flukes removed, washed and rapidly frozen in liquid nitrogen. The content of key metabolites in the fermentation pathway were determined and compared with those in control F. hepatica, whose hosts were not treated with rafoxanide.

Rafoxanide decreased glycogen, malate, NADH and ATP levels. The level of other metabolites in the pathway increased for the first 8–16 h after rafoxanide treatment. The marked decrease in ATP and glycogen, and the increase in total [NAD+]/[NADH] and [oxaloacetate]/[malate], together with the changed content of other metabolites, led to the conclusion that the mode of action of rafoxanide against F. hepatica in vivo is by uncoupling oxidative phosphorylation.

The development of resistance by the Houghton strain of Eimeria tenella to the anticoccidial drugs amprolium, clopidol and methyl benzoquate has been studied. Resistance to amprolium and clopidol developed more readily in experiments where a large number of coccidia were exposed to the drug, either by increasing the number of oocysts in the inoculum or by increasing the number of birds in the group. When 45 birds were given 2·0 × 106 oocysts, resistance to amprolium and clopidol appeared after 6 and 7 passages respectively. In previous experiments, under similar conditions, resistance to robenidine developed after 6 passages, suggesting little difference between these three drugs. Resistance to amprolium and clopidol arose gradually as the concentration of drug was increased, but resistance to methyl benzoquate appeared in a single step from sensitivity to high-level resistance. Both amprolium and clopidol-resistant lines showed an 8-fold reduction in drug sensitivity. Attempts to measure the degree of resistance by calculation of the ED50 were unsuccessful.

An investigation of the carbohydrate energy metabolism of Rhabdias bufonis, the lung-dwelling nematode parasite of the African toad, Bufo regularis, indicates that the nematode stores very little glycogen (0·137 ± 0·003% on a fresh weight basis) but does utilize oxygen in vitro. The intracellular distribution and high levels of activity observed for the enzymes phosphoenolpyruvate carboxykinase, pyruvate kinase, lactate dehydrogenase, malate dehydrogenase, malic enzyme and fumarate reductase suggest two alternative pathways of carbohydrate energy metabolism.

The fine structure and development of the endogenous stages of an embryo-adapted strain of Eimeria tenella in chicken embryos is described. First-generation merozoites were formed by endodyogeny in uninucleate developmental stages as well as schizogony and the endoplasmic reticulum of merozoites of the second-generation schizonts appeared as a circular structure, often enclosing other organelles. In contrast to the parent strain, development of the second-generation schizonts was restricted to cells of epithelial origin. These observations confirm that the complete endogenous cycle of the embryo-adapted strain of E. tenella is restricted to epithelial cells of the chorio-allantoic membrane (CAM) and that no major ultrastructural changes have occurred as a result of repeated embryo passage.

Rats were treated with betamethasone at different stages of infection with Strongyloides ratti. Treatment caused a diminution in the number of adult worms recovered but this residual population was not expelled as were worms from untreated rats. If treatment was started after expulsion had already begun adult worm numbers increased – suggesting that auto-infection occurred. Treatment throughout a primary infection prevented the development of an immune response which expelled, or killed, a challenge infection. Treatment, just before and just after challenge, prevented the expulsion of the challenge infection. Worm expulsion up to day 35 was prevented by treating rats with betamethasone 4, 6, 8 and 10 days after infection.

An in vitro investigation has been made of the relative roles of the gut and tegument in the absorption of the neutral amino acids L-alanine and L-leucine by a marine fish-gill parasite, Diclidophora merlangi. The use of ligatures to preclude oral ingestion of trace-labelled medium has proved inadequate, invariably damaging the tegument, as revealed by stereoscan electron microscopy, and resulting in artifactual levels of absorption. Three alternative procedures have given consistently reliable data on the route of entry of low molecular weight substrates. (1) Ultrastructural examination of worms previously incubated in electron-dense cationic tracers has shown that, in vitro, there is no oral intake of sea water. (2) The suspending of worms in trace-labelled medium with the mouth out of the medium and comparing amino acid uptake with that of worms totally immersed in medium has revealed no statistically significant difference in the absorption levels. (3) Application of section (freeze-dried) auto-radiography to detect diffusible isotope has demonstrated directly transtegumental absorption of a neutral amino acid. It is concluded from these experiments that Diclidophora has a tegumental transport system for absorbing certain neutral amino acids, and whilst, clearly, the worm is sanguinivorous and digests blood in a well-developed gut, it may also be capable of supplementing this diet with low molecular weight organic nutrient absorbed directly from sea water via the tegument.

The cytotoxicity of normal rat spleen cells to antibody-coated Trypanosoma cruzi epimastigotes has been studied by assaying the release of [3H]-labelled macromolecules from the parasites. The release of thymidine (DNA) is slower than the release of uridine (RNA), suggesting that the nucleus is broken down more slowly than the cytoplasmic membrane. Less than 50% of the leucine (protein) is released when the parasites are lysed, whereas uridine (RNA) is almost totally released. In practical terms these results show that the release of incorporated radioisotope-labelled uridine can be used as a sensitive assay for cytotoxicity of T. cruzi. Cytotoxicity by normal rat spleen cells is antibody dependent and proportional to the logarithm of effector cell number. The lag phase and the rate of RNA release is not altered by centrifuging the parasites and effector cells to enhance contacts between them.

Gametogenesis in Plasmodium gallinaceum involves bicarbonate-dependent processes and requires a continuous supply of glucose (presumably as an energy source). Emergence and exflagellation of gametocytes, in vitro, occur independently of the CO2 tension but are rigidly correlated with the pH of the external medium. In bicarbonate-saline gametogenesis is initiated only if the pH exceeds 7·7. Our results suggest that gamete development of malaria parasites is stimulated when infected blood is exposed to air because the decrease in the CO2 tension of the blood causes its pH to rise.

Rediae of Cryptocotyle lingua were studied using transmission and stereoscan electron microscopy. The mouth is surrounded by numerous ciliated sensory structures. Posteriorally, the body wall is concentrically folded and its surface area is greatly increased by the closely ridged configuration of the outer membrane of the tegument. Attached to the surface, especially at the anterior end, are numerous, small spherical vesicles. At the posterior end there is a retractable papilliform process. The appearance of the surface of the birth papilla is variable. The birth pore is lined by an epithelium which is specialized to withstand extreme distortion. A single cilium is frequently visible in the birth pore lumen. The release of each cercaria is preceded by the emission of a tiny drop of fluid from the birth pore. It is suggested that this fluid acts as a lubricant to the passage of the cercaria through the pore.

Mice which have recovered from infections with the avirulent piroplasm Babesia microti are also resistant to challenge with the virulent malaria parasite Plasmodium vinckei. In mice infected with P. vinckei before the peak of the B. microti infection the numbers of malaria parasites in the blood increase until that peak and are then eliminated at the same time as the piroplasms. In mice infected with P. vinckei at or after the peak there is no apparent multiplication and the malaria parasites begin to disappear from the blood immediately. The malaria parasites in doubly infected mice show signs of degeneration similar to those seen in mice pre-treated with Corynebacterium parvum and it is suggested that a common mechanism exists in homologous and heterologous immunity and in immunity following pre-treatment with C. parvum or BCG.

The successful cultivation of a variety of haemoflagellates in three different liquid media is reported. These media include medium 199, Grace's insect tissue-culture medium and Schneider's drosophila medium, each in combination with 30% (v/v) foetal calf serum. These media were used to cultivate Old and New World species of visceral and cutaneous human Leishmania, as well as Leishmania species isolated from sandflies, rodents, and reptiles. Four strains of Trypanosoma cruzi, an isolate of T. rangeli and an isolate of T. lewisi have also been cultivated in these media. One or more of these media have been used to cultivate 121 strains of haemoflagellates, including at least 14 different species (11 Leishmania and 3 Trypanosoma) and many geographic isolates or strains. The Leishmania include L. braziliensis, L. peruviana, L. mexicana, L. tropica, L. donovani, L. chagasi, L. enriettii, L. hertigi, L. hoogstraali, L. adleri, and L. agamae. Using the Schneider's based medium, we have obtained primary isolates of both cutaneous and visceral Leishmania of man and of experimentally infected laboratory rodents and canines. Freeze-dried preparations of the Schneider's based medium that were reconstituted with distilled water after 24 months of storage at ambient temperature have proven to be suitable cultivation media. This feature makes the media valuable field tools.

The various species of human Leishmania cultivated in these media have in our experience demonstrated no differences in growth rate, viability after liquid nitrogen preservation, or infectivity for laboratory animals and tissue-culture cells compared with promastigotes derived from blood-agar cultivation.

Two lines of guinea-pigs, whose susceptibilities to infection with the parasitic nematode Trichostrongylus colubriformis differ significantly, have been established. This difference is probably based on genetically determined differences between the ability of members of each line to bring about immune expulsion. of the parasite.

Fresh (3 h) schistosomula of Schistosoma mansoni were obtained by the skin penetration method and injected intravenously into groups of mice either immediately or after 48 h incubation in tissue culture. Schistosomula were then recovered from the lungs 1 to 2 days after injection. The recovery results showed that the ability of uncultured schistosomula to emerge from lung tissue increased with time, became maximal on the 3rd day and diminished thereafter. In contrast, cultured schistosomula were able to emerge from chopped lung at a much earlier time after injection.

Fresh schistosomula, as well as those recovered from mouse lungs, were assayed by indirect immunofluorescence for the presence of parasite and host antigens. Similarly, fresh schistosomula which had been incubated in foetal calf serum, with or without mouse red blood cells, were assayed for parasite and host antigens after 3–24 h of culture. Fresh schistosomula expressed parasite antigen with greater intensity than those recovered from mouse lungs and showed no trace of host antigen. Lung schistosomula, on the other hand, expressed less parasite antigen but more red blood cell antigen with increasing time in the host. Schistosomula cultured in the absence of mouse red blood cells expressed only parasite antigen throughout the period in culture, while those cultured in the presence of mouse red blood cells also expressed parasite antigen during the entire period in culture but, in addition, by 24 h host antigen could also be detected.

These developmental and adaptational changes may play a role in determining the survival of parasites in the normal or immune host.

Infection of rats with 2000 infective juveniles of Nippostrongylus brasiliensis and of lambs with 60000 infective juveniles of Nematodirus battus results in a well-marked immunity to these nematodes in their respective hosts. There is a fall in the adenylate energy charge value of these nematodes during the course of these infections, reaching values of 0·37 in males and 0·27 in females of N. brasiliensis, and 0·31 in males and 0·23 in females of N. battus towards the end of the infections. In hosts given relatively small numbers of infective juveniles, the values for the nematodes removed from the hosts late in the infection remain at a relatively high level. These results indicate that the immune response of the host may affect the energy status of these nematodes, and this could help to explain their subsequent expulsion from the immune host.