A group of monoclonal antibodies (MAb) was used to immunochemically characterize Babesia bigemina surface components. Two surface reactive MAb by an IFAT test were subsequently shown to bind to epitopes on the external surface of the parasite plasma membrane as evidenced by immunoelectron microscopy. Parasite components with relative sizes of 68, 62, 60, 58, 56, 54, 51, 49, 48, 47, 43, 36 kDa were identified with the group of MAb in at least 6 geographically different B. bigemina isolates. The antigenic components were demonstrated to be species-specific and apparently predominant. Both polyclonal antibodies (immune bovine serum) and highly diluted MAb specifically reacted with such bands producing a strong signal in Western blots.

Hybrid proteins containing selected regions of the major surface antigens of the sporozoite and merozoite stages of Plasmodium falciparum were expressed in insect cells using baculovirus vectors. A recombinant protein containing the signal peptide from the precursor to the major merozoite surface antigens (PMMSA) fused to a fragment from the carboxy (C) terminus of the same gene was recognized by monoclonal antibodies specific for reduction-sensitive conformational epitopes within the C-terminal fragment, suggesting that correct disulphide cross-linking of cysteine residues within this region had occurred. Addition of 26 copies of the tetrapeptide repeat from the circumsporozoite protein (CSP) resulted in a protein recognized by anti-CSP antiserum as well as the conformation specific inonoclonal antibodies. Deletion of the C-terminal putative anchor sequence from both proteins resulted in secretion of protein in a fully soluble form antigenically indistinguishable from the anchor containing products. Correct conformation was not observed when the proteins were expressed as polyhedrin fusions without the signal peptide. These data indicate that the PMMSA signal peptide is recognized in insect cells and that correct assembly of disulphide cross-links is dependent upon targeting the protein to the endoplasmic reticulum.

It was observed that uninfected red cells resuspended in supernatant from Plasmodium falciparum cultures, then examined between a glass slide and cover-slip, assumed varying morphologies. A series of experiments suggested that P. falciparum releases molecules which cause red cells to become stomatocytic (cupped). These molecules, some of which are heat- stable, have an apparent molecular weight < 12 kDa, are released at or about schizogony, and do not bind tightly to erythrocytes.

Blood-stage malaria parasites in the vertebrate host can develop either into the asexual, multiplying forms, called schizonts, or into gametocytes, the sexual stages of the parasite. In the present work we studied the differentiation into asexual parasites or gametocytes of the progeny of single, isolated schizonts of the clone 3D7A of Plasinodium falciparum, using monoclonal antibodies specific for the sexual or asexual stages of the parasite. We observed that schizonts obtained from a continuous culture undergoing serial cycles of growth and dilution with fresh red blood cells produced either only gametocytes or only asexual parasites, showing a high degree of commitment to one or the other developmental pathway.

The relative proportion of schizonts which produced gametocytes was very low at low parasite densities in culture, while at high parasite densities a much greater proportion of schizonts produced gametocytes. Nevertheless, at both low and high parasite densities individual schizonts were almost always fully committed to producing only gametocytes or only asexual parasites.

L-Amino acid esters, such as L-Leu-OMe, kill Leishmania amazonensis amastigotes by a mechanism which appears to involve ester hydrolysis by cysteine proteinases located in the parasite megasomes. We have examined the killing of isolated amastigotes by L-dipeptide esters and derived some structure-activity correlations. Toxicity of the compounds for the parasites was measured by a tetrazolium (MTT) reduction assay. The results show that active dipeptide esters contained at least I hydrophobic amino acid (Leu, Ile, Val, Phe or Trp). The activity of homodipeptide methyl esters depended on the nature of the amino acid, as indicated by the following series: Phe-Phe-OMe < Val-Val-OMe < Leu-Leu-OMe < Trp-Trp-OMe < Ile-Ile-OMe. The nature of the amino acids in Leu-X-OMe and X-Leu-OMe was relatively unimportant when X was Phe, Trp or Val. However, when X was Ala or Gly, Leu-X-OMe was several-fold more active than X-Leu-OMe.

A simple model of human and animal trypanosomiasis is proposed in which the Ross equation for disease transmission is supplemented by a differential equation describing the inheritance of susceptibility in the vector. The model predicts an equilibrium state of balanced polymorphism for the fraction, θ, of susceptible tsetse and the occurrence of periodic epidemics at roughly the observed intervals. A loss of infectivity to tsetse of mechanically transmitted strains of trypano some would seem to be a good evolutionary strategy for the trypanosome. The main implication for disease control is that measures initially reducing trypanosomiasis incidence could trigger off subsequent epidemics. Since θ leads incidence, monitoring θ could give several years advance warning of major epidemics.

Numerical taxonomy was used to review isoenzyme variation in isolates of Trypanosoma brucei obtained from cattle, tsetse, humans and wildlife from the Lambwe Valley, Kenya. From isoenzyme information alone, it was possible to classify isolates as to source through the use of linear discriminant functions analysis, with an error rate of only 2% in humans, and 14% over all groups. Differentiation was mostly dependent on patterns in the enzymes ASAT, PEPI, and ICD. Parasites from non-human sources, especially tsetse, were characterized by high isoenzyme diversity, and many unique zymodemes. Observed frequencies of genotypes for ICD, ALAT, and ASAT did not agree with expected frequencies based on random mating of a diploid organism. Deviations were particularly large for tsetse isolates, and were mostly due to a deficiency of one homozygote. Cluster analysis revealed complex relationships among isolates, with patterns evolving through time. Major human zymodemes from the 1970s clustered together with most wildlife isolates from East Africa. This chronic human-wildlife transmission cycle was characterized by ASAT pattern I. Other, minor human zymodemes were associated with a human-cattle transmission cycle characterized by ASAT pattern VII. These original chronic transmission cycles appeared to change in 1980 with the appearance of two new zymodemes in humans. These zymodemes involved changes in ALAT and/or PGM to patterns typical of tsetse and cattle isolates. A resultant epidemic was halted with repeated aerial spraying of endosulfan in 1981. Since then, a variety of new zymodemes of unknown human infectivity have appeared. The origins of these changes are discussed in terms of genetic exchange in tsetse, adaptation to human and cattle transmission cycles, and selection resulting from chronic use of insecticides.

Sera from rabbits, rats and mice multiply-vaccinated with attenuated cercariae of Schistosoma japonicum conferred high levels of resistance against challenge to naive recipient mice (up to 97, 64 and 60% respectively). Vaccinated rabbit and rat sera were given before challenge and vaccinated mouse serum 5 days after challenge. To show that the protective factors in these sera were antibodies, vaccinated rabbit and mouse sera were fractionated by protein A-Sepharose and the fractions precipitated by 50% ammonium sulphate. The protein A-Sepharose binding or non-binding fractions in vaccinated rabbit serum transferred approximately equal levels of significant resistance to mice, suggesting that both the IgG and non-IgG components of vaccinated rabbit serum are protective. The major part of the protective activity in vaccinated mouse serum was transferred to recipients by the protein A-Sepharose binding fraction, i.e. the IgG antibodies. Heat inactivation of sera at 56 °C for 3 h affected the protective capacity of vaccinated rat sera, but not that of vaccinated rabbit or mouse sera.

Deletion and/or amplification of DNA sequences in Schistosoma mansoni were demonstrated by Southern blot analysis. Total cellular DNAs and genomic clones derived from S. mansoni miracidia, adult males and females were used as probes. EndonucleaseBamHI-restricted DNAs from miracidia, adult males and females of both S. mansoni and S. japonicum were reacted to each probe. Hybridization with a total cellular DNA from S. mansoni miracidia as a probe showed elimination of signals in S. mansoni adults. On the other hand, blot analysis using a total cellular DNA from S. mansoni adult males as a probe revealed elimination of hybridization signals in S. mansoni miracidia.Hybridization with a clone SmE15 DNA from S. mansoni miracidia as a probe showed no signal in the DNAs from 5. mansoni adults, indicating these sequences deleted in adults. Hybridization experiments using the probes SmF25 and SmM51 which are 13 and 2-2 kb fragments cloned from S. mansoni adult females and males respectively, demonstrated no signal to DNA from S. mansoni miracidia.Our data suggested the existence of stage-specific DNA sequences in 5. mansoni. We propose a model for multiple-step rearrangement of DNA sequences in S. mansoni during the course of development.

The prevalence of patent schistosome infections of intermediate host snails often shows seasonal variations. For schistosome, mainlySchistosoma haematobium, infections of Bulinus globosus field data from Zimbabwe show annual ranges in prevalence from 2% to 30%. In this paper a mathematical model is developed for B. globosus population dynamics and the epidemiology of schistosome infection. The model is a discrete-time analogue of a catalytic model, with an added recruitment term. Snail fecundity and survival are functions of age, infection, and temperature. The pre-patent period of infection is dependent on temperature.Parameter values, and relationships with temperature, are taken from earlier field and laboratory studies. The force-of-infection is estimated from analysis of age-prevalence data. Model predictions, making use of temperature data recorded in the field, are in good agreement with observations over a 2-year period at a study site in Zimbabwe. The key features of interest are a fall in the prevalence of patent infections during the cold season and a rapid rise to a peak early in the hot season. This pattern results from the accumulation of pre-patent infections which fail to develop at low temperatures, but mature en masse, together with new infections, after a few weeks at higher temperatures. Model analysis demonstrates that seasonal patterns in the prevalence of patent infections may be due largely to the influence of temperature on the pre-patent period. Seasonal and year-to-year variations in temperature may therefore have significant epidemiological effects.

Confocal scanning laser microscopy has been employed with immunocytochemical techniques to map the distribution of serotoninergic and peptidergic components in the nervous system of the monogenean gill-parasite, Diclidophora merlangi; results are compared with the distribution of cholinergic components, following histochemical staining for cholinesterase activity. While all three neurochemical elements are present in the central and peripheral nervous systems, the cholinergic and peptidergic systems dominate the CNS, whereas the PNS has a majority of serotoninergic nerve fibres. The cholinergic and peptidergic neuronal pathways overlap extensively in staining patterns, suggesting possible co-localization of acetylcholine and neuropeptides. Within the peptidergic nervous system, immunoreactivity to the pancreatic polypeptide family of peptides and FMRFamide were the most prevalent.

The time taken for larvae of Nippostrongylus brasiliensis and a homogonic strain of Strongyloides ratti to complete their migration from the skin surface to the small intestine of the rat was estimated by a new method in which a mirror image of arrival times was created by counting mature parasites at day 8 (N. brasiliensis) or day 5 (S. ratti) in the intestines of rats that had received a single pulse of morantel tartrate by stomach tube at different times starting 10 h after skin application of exact doses of infective larvae. It was confirmed that the effect of the drug was confined to parasites in the gut, so that the kinetics of migration were unperturbed.

Rigorous proofs applicable to the routes of migration of Strongyloides ratti and Nippostrongylus brasiliensis skin-penetrating juveniles inside the rat are extended. By applying the inequality principle (Tindall & Wilson, 1988) it was confirmed with a probability of error of I in 1010 that N. brasiliensis larvae applied to the skin passed through the lungs on their way to the intestine. Taking the analysis further, migrating larvae of S. ratti or N. brasiliensis were extracted from the nose or lungs, respectively, of donor rats and transferred to recipients by stomach tube to assay their ability to colonize the intestine. Results showed that (a) changes undergone by each parasite in its proven, specific transit site were essential before larvae could establish in the intestines of recipients, (b) these changes were not completed until larvae had been in the nose or lung for a significant period.

The presence of entomopathogenic nematodes in soil from 15 sites on 10 sampling occasions over a period of 28 months was documented. Sites were chosen to represent habitats classified as heathland, hedgerow, roadside verge, pasture or woodland. Bioassays utilized Galleria larvae and most of the nematodes obtained were identified asSteinernema (= Neoaplectana) bibionis (Bovien). A sibling species which fails to interbreed with S. bibionis was also found at 2 of the sites. Bioassays were at 15 and 20°C and there was no significant difference in the results at these temperatures. There was no evidence for seasonality in the presence of the nematodes and at 2 sites they were found on every sampling occasion. Otherwise, sites converted unpredictably from positive to negative or negative to positive over the 28-month sampling period.

Soil from 403 hedgerow, roadside verge, woodland, heathland or field sites was assessed for presence of entomopathogenic rhabditid nematodes with aGalleria larva baiting technique. Steinernematids were recovered from 48 % of the sites but only one site yielded Heterorhabditis sp. The species recovered most frequently was Steinernema bibionis (Bovien). Another unidentified Steinernema sp. was also isolated. Prevalence of steinernematids in the different types of habitat formed continuum so that roadside verges harboured them most often while heathland sites yielded them least often. Mean soil temperatures at 5 cm were about 15 °C in fields and verges, 13 °C in hedgerows and heathland and 12 °C in woodland. Prevalence of the nematodes varied in different parts of Britain and was influenced by soil type. They were associated with calcareous soils, especially those with a calcareous subsoil horizon. Soils which are periodically or seasonally water-logged were suitable. Soils enriched by humus and hence high in organic matter frequently harboured the nematodes and some suitable soils had significant clay contents or subsoils high in clay. It appears that the two steinernematids are relatively unspecialized species, occurring in diverse habitats and soil types and are adapted to cooler temperatures.

Intensity of Ascaris lumbricoides infection was measured in terms of egg counts and worm burden in children 2–10 years of age. The expulsion of A. lumbricoides with a 3-day treatment of mebendazole occurred over 8 days, beginning on the second day of treatment. Ninety-seven percent of the worms were expelled between the second and seventh days. A rapid means of estimating eggs per gram (epg) by the Kato Katz technique correlated well with the method described by Martin & Beaver (1968). In spite of apparent density dependence in egg production, A. lumbricoides egg counts correlated well with worm burdens. It is concluded that, within the context of community surveys, epg is a reasonable means of identifying heavily infected individuals and that epg can be estimated rapidly by a slight modification to the standard Kato Katz technique.

Killing of metacestodes by normal or post-infection macrophages and the regulation of this activity by cytokines were studied in vitro. The protoscolecidal activity of normal macrophages against Echinococcus granulosus was inhibited by a product of naive T-enriched lymphocytes co-cultured with protoscoleces (PSC). By contrast, supernates from co-cultures of Mesocestoides corti tetrathyridia (MCT) and T-enriched or B-enriched normal lymphocytes increased killing of MCT by normal macrophages. Larvicidal activity (against both PSC and MCT) was enhanced by high concentrations of macrophage-activating factors produced by Con A-stimulated rat lymphocytes (Con A-LK), but was reduced by low concentrations of these factors.

Baseline data from an immuno-epidemiological study of hookworm infection in a rural village in Madang Province, Papua New Guinea are reported. Necator americanus was found to be the commonest helminth infection, with a prevalence of near 100% and intensity of 40 worms per host in adults. Enterobius vermicularis, Ascaris lumbricoides and Trichuris trichiura were also present, at prevalences of 53, 10 and 3% respectively; Ancylostoma duodenale was absent. The frequency distribution of N. americanus was highly over-dispersed, and was well described by a negative binomial distribution with aggregation parameter, k, of 0·370. Intensity of infection was significantly related to host age, but did not differ between the sexes. Haemoglobin levels and haematocrit values were indicative of anaemia in the community, but were unrelated to hookworm infection. Levels of antibodies (IgG, IgA and 1gM combined) against adult Necator cuticular collagen and excretory-secretory (ES) products were determined.

A Strain ofHaemonchus contortusshowing moderate levels of resistance to thiabendazole (TBZ) (egg hatch assay LC50 2.1 μM) was subjected, in alternating sequence, to 5 in vitro and 4 in vivo selections with TBZ. This resulted in a significant increase in resistance (egg hatch assay LC50 2.8 μM). The isolates obtained after each combined in vitro and in vivo selection were compared in terms of egg hatch assay, establishment, net egg output, per capita egg output and degree of pathology caused in infected hosts. The increase in resistance was accompanied by a significant increase in larval establishment sheep (22 %) and was associated with more severe pathology, higher egg count and worm recovery 3 months post- infection.

Antineoplastic cyclophosphamide (CY) significantly reduced the survival and the weight of Hymenolepis microstoma in mice. Corticosteroid dexamethasone (DEX), on the other hand, did not influence the worm survival and the worms in DEX-treated mice became significantly heavier. Both immunosuppressants abolished H. microstoma-induced bile duct hypertrophy and submucous enteritis and lowered the number of mast cells (MC) in these regions. CY appeared to be more effective in depleting the bile duct MC whereas DEX seemed to affect more the duodenal MC population.