An in vitro system is described in which over 85% of a population of Leishmania mexicana mexicana amastigotes transforms to promastigotes within 48 h. The differentiation process involves 3 morphologically and biochemically distinct intermediates, including a division stage. Cell division is shown to be necessary for complete development to promastigotes. Foetal calf serum (FCS) has been found to be an essential component of the medium for high percentage transformation to be achieved. One of the important components of the FCS has been identified as non-esterified fatty acids, and these support a relatively high percentage of amastigotes through transformation in the absence of FCS, possibly due to their use as energy substrates. Only small numbers of amastigotes transform to promastigotes if glucose or amino acids are the only energy substrates available. Transformation is inhibited by a number of metabolic inhibitors including anti-leishmanial and other anti-protozoal drugs. The stage at which inhibition is apparent varies with the inhibitor. It is suggested that the system described for the transformation in vitro of L. m. mexicana amastigotes to promastigotes may be the best method available at present for studying the metabolism and drug sensitivity of amastigotes free from possible interference by host macrophage components.

Rainbow trout were infected simultaneously with cystacanths of the two acanthocephalan species Pomphorhynchus laevis and Acanthocephalus anguillae in the laboratory. Infections consisted of varying proportions of the two species (from 0 to 100%) at low (20 cystacanths) and high (60 cystacanths) levels. Fish were sacrificed at 7, 56 and 112 days post-infection (p.i.) and the parasites recovered. At low-level infections there was a high degree of overlap in the range of intestine occupied by the two species (44·12 %) and no evidence for interspecific competition affecting either species. At high-level infections the establishment of both species was unaffected by the presence or number of individuals of the other species. The survivorship and the range of intestine occupied by A. anguillae were found to be reduced in the presence of P. laevis. The interaction was one-sided, as P. laevis remained unaffected by A. anguillae, and so indicated interference competition. The possibility that both intraspecific and interspecific competition are implicated in explaining the distribution of the two species in the British Isles is discussed.

Immune responses of 4 inbred lines of chickens, that differ in resistance to Eimeria maxima and E. tenella, were examined. Significant differences were found in in vitro proliferation of peripheral blood lymphocytes to E. maxima sporozoite antigen, the more resistant lines C and 72 having higher responses than the more susceptible line 151. These differences existed pre-infection and were enhanced following both primary and a second infection. The proportions of lymphocyte subsets in the peripheral blood following primary infection also differed between lines, with significantly higher percentages of CD8 + and TCR1 + lymphocytes circulating in the more resistant birds. In contrast, there were few differences between lines in either resistance or in in vitro proliferation of peripheral blood lymphocytes to E. tenella sporozoite antigen either pre-infection or following a primary infection. However, after a second infection when there were significant differences in resistance between lines, as measured by oocyst excretion, there were also significant differences in lymphoproliferation with the more resistant lines 151 and 62 having higher responses than the more susceptible line C. Thus for E. maxima there is a direct relationship between resistance to infection and lymphoproliferation in response to parasite antigen. This implies that differences in cellular immunity may account for differences in resistance between lines, and since these specific responses are enhanced by infection they may also reflect important immune mechanisms. For the rather less immunogenic E. tenella, the correlation between resistance and lymphoproliferation is not so clear. However, where there were significant differences between lines, i.e. after a second infection, the direct relationship between resistance and lymphoproliferation was upheld.

Despite important differences between infectious diseases and cancers, tumour development (neoplasia) can nonetheless be closely compared to infectious disease because of the similarity of their effects on the body. On this basis, we predict that many of the life-history (LH) responses observed in the context of host–parasite interactions should also be relevant in the context of cancer. Parasites are thought to affect LH traits of their hosts because of strong selective pressures like direct and indirect mortality effects favouring, for example, early maturation and reproduction. Cancer can similarly also affect LH traits by imposing direct costs and/or indirectly by triggering plastic adjustments and evolutionary responses. Here, we discuss how and why a LH focus is a potentially productive but under-exploited research direction for cancer research, by focusing our attention on similarities between infectious disease and cancer with respect to their effects on LH traits and their evolution. We raise the possibility that LH adjustments can occur in response to cancer via maternal/paternal effects and that these changes can be heritable to (adaptively) modify the LH traits of their offspring. We conclude that LH adjustments can potentially influence the transgenerational persistence of inherited oncogenic mutations in populations.

1. The seasonal incidence of Nematodirus infection in sheep was followed by faecal egg counts, post-mortem examination and pasture larval studies.

2. It was shown that only one generation of adult worms was produced each year and that the life history required a prolonged resting stage on the pasture, which took 12 months to complete.

3. The influence of this type of life cycle on the parasitic disease caused by Nematodirus, and on its control, are discussed.

Parasitism by the flea Synosternus cleopatrae does not affect the body mass of its principal rodent host, Gerbillus andersoni under natural infestation levels. We hypothesized that the lack of negative effects of flea parasitism on rodent body mass could be related either to the low level of natural infestation or to the differential susceptibility of rodent age cohorts to flea parasitism. We tested these hypotheses by measuring body mass change under flea parasitism in (a) adult rodents infested with fleas above the natural infestation level (the first hypothesis) and (b) juvenile rodents infested with fleas at natural infestation levels (the second hypothesis). Adult individuals parasitized by a number of fleas higher than in nature lost body mass at higher rates than non-parasitized control individuals. Parasitism significantly affected daily body mass change of juvenile gerbils. Juvenile rodents parasitized by fleas at the natural level of infestation lost body mass faster and gained body mass slower than control animals. We suggest that some regulating mechanisms may limit natural flea densities at a point at which the negative effect on hosts is below the accuracy of our measurements. However, natural flea densities are sufficiently high to harm the more susceptible, juvenile cohort.

Cathelicidins are antimicrobial peptides produced by humans and animals in response to various pathogenic microbes. Crotalicidin (Ctn), a cathelicidin-related vipericidin from the South American Crotalus durissus terrificus rattlesnake's venom gland, and its fragments have demonstrated antimicrobial and antifungal activity, similarly to human cathelicidin LL-37. In order to provide templates for the development of modern trypanocidal agents, the present study evaluated the antichagasic effect of these four peptides (Ctn, Ctn[1-14], Ctn[15-34] and LL-37). Herein, Ctn and short derived peptides were tested against the epimastigote, trypomastigote and amastigote forms of Trypanosoma cruzi Y strain (benznidazole-resistant strain) and cytotoxicity in mammalian cells was evaluated against LLC-MK2 lineage cells. Ctn inhibited all T. cruzi developmental forms, including amastigotes, which is implicated in the burden of infection in the chronic phase of Chagas disease. Moreover, Ctn showed a high selective index against trypomastigote forms (>200). Ctn induced cell death in T. cruzi through necrosis, as determined by flow cytometry analyses with specific molecular probes and morphological alterations, such as loss of membrane integrity and cell shrinkage, as observed through scanning electron microscopy. Overall, Ctn seems to be a promising template for the development of antichagasic agents.

Guidelines recommend praziquantel (PZQ) for the treatment and control of schistosomiasis, with no real alternative. Metrifonate was still widely used against Schistosoma haematobium in the 1990s, and then withdrawn. Experimental studies and clinical trials suggest that artemisinin compounds are active against S. haematobium. In a Cochrane systematic review assessing the efficacy and safety of drugs for treating urinary schistosomiasis, 24 randomized controlled trials (n=6315 individuals) met our inclusion criteria. These trials compared a variety of single agent and combination regimens with PZQ, metrifonate or artemisinin derivatives. The review confirmed that both the standard recommended doses of PZQ (single 40 mg/kg oral dose) and metrifonate (3×7·5–10 mg/kg oral doses administered fortnightly) are efficacious and safe in treating urinary schistosomiasis, but there is no study comparing these two regimens head-to-head. There is currently not enough evidence to evaluate artemisinin compounds. Most of the studies included in the Cochrane systematic review were insufficiently powered, lacked standardization in assessing and reporting outcomes, and had a number of methodological limitations. In this paper we discuss the implications of these findings with respect to public health and research methodology and propose priority research needs.

Traditionally, parasitology has been concerned with the harmful effects of parasitic organisms; it is basically an applied science. Since its founding in the field of medicine, and later in veterinary medicine, parasitology has been mainly devoted to generating knowledge, which is applicable to parasite control and management, and eventually to their eradication. However, the complexity of parasitism, as revealed over recent decades by workers in various specialisms of parasitology, makes the application of management and control measures very difficult in natural environments. This is particularly true in the marine realm, where however some applied aspects of parasitology, other than those devoted to control and management of parasites, have been shown to be of great importance for fisheries, human health, biological control of introduced species and environmental sciences (Rohde, 2002). The relationship of parasitology with mariculture practices, which are carried out mostly on artificial and controlled systems, follows a more classical approach, focusing on the development of strategies of prevention, management and control of pathogens.

1. The growth and development of normal and sterile (i.e. symbiont-free) Rhodnius prolixus are compared. The last three instars of sterile bugs weigh less than normal ones. Sterile insects completed the first three stages of their life-histories in less time than normal insects, and at the fourth stage, nutritional deficiencies had profound effects on their further development.

2. The sensitivity of the Rhodnius symbionts, Nocardia rhodnii, to five antibiotic substances is determined. Terramycin, aureomycin and chloramphenicol are effective inhibitors.

3. The symbiotic condition in Rhodnius and the related bug, Triatoma infestans, is discussed.

The immunological differences known to exist between laboratory strains of Eimeria maxima was confirmed. Protection against challenge with different strains or field isolates of the species could be achieved by including small numbers (25 oocysts) of each in the immunizing inoculum. Similar protection was obtained when 4 distinct populations which were allowed to interbreed were used in the immunizing inoculum. This hybrid mixture of E. maxima was used to immunize chickens against challenge with 7 new isolates of E. maxima from poultry houses in different parts of England. The results show that although immunological differences exist within E. maxima good protection against many strains of this species may be achieved by initial infection with the hybrid mixture of E. maxima.

Infective larvae, adults and newborn larvae of Trichinella spiralis were surface labelled with radioactive iodine, and the surface material was solubilized in the mild detergent sodium deoxycholate. The radio-isotope labelled products were stage-specific glycoproteins that were few in number (2–4 components) and antigenic in infected mice and rats. Antibodies synthesized in infected animals against these biochemically defined surface antigens may or may not interact with the surface of the living worm. The latter type of antibody is unlikely to be involved in the initial phase of parasite rejection and is therefore another example of a non-protective host antibody response. The stimulus for its synthesis must be the observed release of surface antigen. A monoclonal antibody to a surface glycoprotein of newborn larvae protected against infection, and also promoted eosinophil killing in vitro. This observation emphasizes the importance of surface antigens in protection against infection, suggests a role for granulocytes in vivo, and provides encouragement for the possible use of nematode surface antigens in protection. An example of regional specialization of the nematode cuticle was given by a monoclonal antibody reactive with only the surface of the male intromittent organ and not the female or remainder of the male. The same stages were labelled in vitro with radioactive methionine, and the secreted proteins were also found to be stage-specific. Some, but not all, were antigenic in infected mice. The total concanavalin A-binding somatic glycoproteins of each stage exhibited considerable individuality, and hence stage specificity, when resolved by two-dimensional gel electrophoresis. These results extend our knowledge of stage-specific components of T. spiralis, and allow a rational approach towards protection and the construction of diagnostic procedures.

Human populations are often infected with more than one species of parasite, especially in developing countries where overall rates of parasitism are high. Infections with multiple parasite species may not necessarily be independent within an individual as physiological, immunological or ecological factors may result in positive or negative associations between infections with different parasite species. A general framework for estimation of these associations is presented. Data from over 215000 individuals are analysed and the associations between geohelminth (Ascaris lumbricoides, Trichuris trichiura and hookworm) and malaria species are investigated. A method is presented for analysing data from multiple communities and testing whether the associations in different communities are equal. Overall estimates of the associations between species are obtained for each country and continent where data were available. Associations between geohelminth species were, in general, found to be positive whilst both positive and negative associations were found between the different Plasmodium species. There was evidence for significant geographical heterogeneity between the associations. A method for using these parameter estimates to predict the distribution of multiple infections when only marginal prevalence data are available is described and demonstrated.

Classical transmitters are present in all phyla that have been studied; however, our detailed understanding of the process of neurotransmission in these phyla is patchy and has centred on those neurotransmitter receptor mechanisms which are amenable to study with the tools available at the time, for example, high-affinity ligands, tissues with high density of receptor protein, suitable electrophysio-logical recording systems. Studies also clearly show that many neurones exhibit co-localization of classical transmitters and neuropeptides. However, the physiological implications of this co-localization have yet to be elucidated in the vast majority of examples.

The application of molecular biological techniques to the study of neurotransmitter receptors (to date mainly in vertebrates) is contributing to our understanding of the evolution of these proteins. Striking similarities in the structure of ligand-gated receptors have been revealed. Thus, although ligand-gated receptors differ markedly in terms of the endogenous ligands they recognize and the ion channels that they gate, the structural similarities suggest a strong evolutionary relationship. Pharmacological differences also exist between receptors that recognize the same neurotransmitter but in different phyla, and this may also be exploited to further the understanding of structure-function relationships for receptors. Thus, for instance, some invertebrate GABA receptors are similar to mammalian GABAa receptors but lack a modulatory site operated by benzodiazepines. Knowledge of the structure and subunit composition of these receptors and comparison with those that have already been elucidated for the mammalian nervous system might indicate the functional importance of certain amino acid residues or receptor subunits. These differences could also be exploited in the development of new agents to control agrochemical pests and parasites of medical importance.

The study of the pharmacology of receptor proteins for neurotransmitters in invertebrates, together with the application of biochemical and molecular biological techniques to elucidate the structure of these molecules, is now gathering momentum. For certain receptors, e.g. the nicotinic receptor, we can expect to have fundamental information on the function of this receptor at the molecular level in both invertebrates and vertebrates in the near future.

The course of primary infection was studied in BALB and B10 H-2 congenic mouse strains. The duration of infection, as assessed with regular faecal egg counts and worm burdens, was shorter in mice carrying the H-2s, H-2d or H-2q haplotypes when compared to mice with H-2b. Strains with H-2k were intermediate. An experiment was carried out to test the hypothesis proposed by Wassom, Krco & David (1987) predicting that the progeny of I–E+ve mouse strains crossed with I–E-ve strains, would show susceptibility rather than resistance to infection. This hypothesis was not substantiated by our data and we conclude that it does not apply to primary infections with Heligmosomoides polygyrus. It is proposed that the gene products of at least two loci within the H-2 (associated with the H-2b and H-2k haplotypes) are crucial in determining the response phenotype of mice to primary infection with H. polygyrus. One allele, (associated with the H-2b haplotype) may be preferentially affected by parasite-mediated immunosuppression.

The host–parasite relationships of Schistosoma mansoni in the albino rat have been studied recently by Ritchie, Garson & Knight (1963) and by Sadun & Bruce (1964). The general findings of these and other workers are that, in rats, the worms are always stunted, although they may pair and non-viable eggs may be found in the liver. The worms remain in the liver and seldom, if ever, migrate to the mesenteries. Ritchie et al. (1963) found that the majority of the worms were eliminated between the 4th and 8th week after infection; this is much sooner than in other experimental hosts. Sadun & Bruce (1964) suggest that rats are particularly suitable and convenient for studies on acquired resistance to S. mansoni; this suggestion is based mainly on the grounds that acquired resistance is more easily demonstrated in animals which show a relatively great degree of innate resistance.

The present work has been undertaken in order to establish whether the immune response in the albino rat is at all similar to that in the rhesus monkey (Smithers & Terry, 1965b). Two features of the host–parasite system have been studied in detail; the elimination of the worms which follows an initial infection and a comparison of the development of acquired resistance following exposure to X-irradiated cercariae with that following exposure to normal cercariae.

A Puerto-Rican strain of S. mansoni, and Sprague–Dawley rats weighing 100–200 g, were used in all experiments. The methods of exposing the rats percutaneously to infection and the recovery of the worms by perfusion have been fully described (Smithers & Terry, 1965 a).

Leishmania (Leishmania) infantum zymodeme MON-1 is responsible for the majority of visceral leishmaniasis cases around the Mediterranean basin, albeit that it causes also cutaneous forms. The MON classification is based on starch gel multilocus enzyme electrophoresis (MLEE) typing. The aim of this work was to explore further the genetic diversity and phylogenetic status of this zymodeme by alternative typing techniques. Fourteen L. (L.) infantum/L. (L.) chagasi stocks identified as MON-1 by MLEE in reference laboratories, 3 L. infantum stocks attributed to other zymodemes (MON-24, MON-29, MON-33) and reference standard stocks belonging to other species (L. (L.) major, L. (L.) tropica and L. (L.) donovani) were characterized by 2 different markers: MLEE on cellulose acetate plates and Random Amplified Polymorphic DNA (RAPD). We have obtained 10 different genotypes with RAPD and 6 different genotypes with MLEE on cellulose acetate plates for the 14 L. infantum/L. chagasi MON-1 stocks studied. MLEE and RAPD data gave quite congruent phylogenetic results: L. infantum zymodeme MON-1 was shown to be polyphyletic and genetically heterogeneous. This work confirms the necessity of using different markers to build up a robust phylogeny. Finally the epidemiological and clinical implications of these results are discussed.

The sex ratio of adult worms has been observed biased towards males in Schistosoma mansoni under natural conditions. The origin of this bias is unknown. This paper determines whether males are more infective than females under controlled experimental bisexual conditions, and hence if the sex ratio is male-biased as a consequence of this. The male and female cercarial infectivities in uni- and bisexual vertebrate host infections using a range of controlled cercarial sex ratios were studied. The results showed that, in experimental unisexual infections, male cercariae were more infective than females, and that in experimental bisexual infections, male cercarial infectivity was similar to that of female, irrespective of cercarial sex ratio. Furthermore, cumulative male and female cercarial infectivity was maximal when sex ratio was equilibrated. The unbiased sex ratios obtained in our experimental bisexual infections are discussed in terms of behavioural and/or biochemical male–female interaction. Alternative explanations of the natural biased sex ratio are proposed.

1.Studies on the histological changes during digestion in Argas persicus reveal that the ingestion of blood is accompanied by the destruction of the existing gut epithelial cells.

2.The blood remains unlysed for 2–3 days while a new epithelium develops which contains cells that secrete a saliva-fast PAS-positive colloid that causes haemolysis.

3.Other epithelial cells remove the freed erythrocytic nuclei by phagocytosis.

4.Most of the gut cells then absorb protein from the lumen and intracellular digestion takes place leaving pure haematin granules as the waste product of the digestion of haemoglobin.

5.After the initial rapid phase of digestion only relatively few cells show signs of absorptive and digestive activity.

6.Absorption of protein is accompanied by increased alkaline phosphatase activity in the microvilli of the cell border; this activity is lost once absorption finishes and digestion begins.

7.Strong aminopeptidase activity can be demonstrated at the border of some of the protein vacuoles.

8.In vitro tests show that the gut proteinase is active only in the acid range with peaks at pH 2·6 and 3·8 and has a Km of 0·32 % with bovine serum albumin as substrate.

9.In vivo data show that digestion, after haemolysis has occurred, takes place in two phases; the first is rapid and lasts approximately 1–2 weeks and is followed by the second phase which is slow and remains constant until the next blood feed.

10.The proportion of the blood meal which remains after the rapid phase of digestion is determined by the sex and developmental stage of the tick and within each category it is constant and serves, in the absence of significant fat and glycogen reserves, as a food reserve.

The snail host signals releasing host-finding responses in miracidia and cercariae of Echinostoma caproni were analysed by fractionation of snail-conditioned water (SCW). Cercariae responded non-specifically to organic and hydrophilic, low molecular weight components of SCW showing their typical turning response. Hydrolysis of peptides in SCW had no effect on cercarial responses. An artificial mixture of amino acids in concentrations determined from SCW as well as glycine alone in a concentration corresponding to the total concentration of amino acids in SCW showed nearly the same efficacy as SCW itself. Miracidia responded to a high molecular weight glycoprotein fraction, which could be isolated from SCW by ion-exchange and size-exclusion chromatography. In contrast to an Egyptian Schistosoma mansoni strain, the echinostome miracidia were not able to differentiate between different snail species. The results show for the first time that miracidia and cercariae of the same species may use different signals to identify the same snail host species. This indicates an independent evolution of host-finding mechanisms in the two parasite stages.