Oocyst prevalence and intensity have been recorded in 349 laboratory infections of Anopheles stephensi with Plasmodium berghei. Intensity and prevalence of infection are shown to be predictably related. The structure and heterogeneity in the infections has been analysed with the objective of describing the biological mechanisms by which the observed negative binomial oocyst distributions are generated. The analysis has revealed that the most likely processes lie within the population dynamic events of malaria within the mosquito, namely gametogenesis, fertilization and mortality. The distribution is similar in all Plasmodium – mosquito combinations examined so far, whether they are of laboratory (P. gallinaceum in Aedes aegypti) or field (P. vivax in An. albimanus and P. falciparum in An. gambiae s.l. and An. funestus) origin. Further we conclude that there is competition between parasites in the vector. Oocyst frequency distribution analysis shows that under natural conditions of transmission intensity, and even under the best laboratory conditions, significant numbers (> 10%) of fully susceptible mosquitoes will not be infected under conditions where the mean infection is as high as 250 oocysts. Failure to infect is not therefore an absolute indicator of refractoriness. In assessing transmission data it is shown that sample sizes should not be less than 50, and ideally 100 mosquitoes, if reliable data are to be obtained. In the field it is suggested that difficulties in determining the low natural intensity of oocyst infections indicate that prevalence estimates are a useful and accessible parameter to measure. In determining the impact of transmission blocking mechanisms we predict that under conditions where high oocyst intensities prevail, large reductions in intensity will be required before a reduction in prevalence can be expected i.e. here it will be necessary to measure intensity of infection. Conversely, under conditions where low oocyst intensities prevail, a rapid reduction in prevalence will occur with little concurrent reduction in intensity i.e. prevalence determination will be the more sensitive estimate.

A recombinant baculovirus-expressed hybrid protein containing epitopes for the C-terminal fragment of the Plasmodium falciparum precursor to the major merozoite surface antigens (PMMSA) and the tetrapeptide repeats of the circumsporozoite protein (CSP) was assessed for its immunogenicity. Murine MHC-II restriction of the antibody response to the CSP repeats was not overcome by the PMMSA component, the response to which showed no restriction. In an adjuvant trial the highest antibody titres in rabbits to both components of the hybrid were obtained using Freund's adjuvant. Lack of a boosting antibody response to the CSP repeats appeared to be linked to the conformation of the PMMSA component. Formulation of the hybrid protein into Iscoms gave antibody titres of only short duration to both components.

Since pigs are important in the zoonotic transmission of schistosomiasis japonica in China, a veterinary vaccine might contribute to the control of the disease in humans. Pigs were immunized with three doses each of 10000 cercariae of Schistosoma japonicum attenuated with ultraviolet light (400 μWatt·min/cm2). The experiment was performed with portable irradiation equipment in a rural area of the Hubei Province (P.R. China). A challenge infection of 1000 untreated cercariae was given 2·5 or 6 months after the last immunization, and age-matched naive pigs were challenged as a control. Immunized pigs developed about 90% resistance against the challenge. The liver egg load of these animals was reduced by over 90%. Less than 0·01 % of the immunizing cercariae developed to adult parasites and the vaccination had no apparent adverse influence on the pigs' health.

Sera from rabbits vaccinated with irradiated cercariae mediated cell (P388D1 or mouse peritoneal macrophage) adherence to lung-stage schistosomula (LS) but such antibody-mediated cell adherence was short-lived in contrast to cell adherence to mechanically transformed schistosomula (MS). Thus LS lost 50% of their adherent cells within 3–6 h in culture and up to 90% by 24 h, whereas adherence to MS was undiminished during this time. Rapid loss of adherent cells was unique to schistosomula that had developed to the lung stage because schistosomula recovered from the skin up to 3 days post-infection did not exhibit the rapid cell loss shown by 3-day LS. To determine whether cell loss was caused by loss of surface antigenicity during culture LS were cultured on their own for up to 24 h and at various intervals samples of schistosomula were tested for antigenicity by addition of immune serum and cells. Levels of adherence to both MS and LS were maintained throughout the incubation period. When antibody-opsonized schistosomula were washed and indicator cells added at progressive intervals, persistence of adherence was again demonstrated, showing that antibody binding to LS had not promoted surface antigen loss or degradation of bound antibody. It was then shown, by adding fresh macrophages to cultures up to 24 h old that LS which had lost their adherent cells nevertheless retained bound antibody, and comparison of adherence of ‘used’ and ‘fresh’ cells to MS and LS showed that the cytoadherence properties of macrophages were not significantly reduced during their culture with LS from which cells had been lost. Cell loss was shown to be dependent upon protein synthesis by LS since cell loss was significantly reduced when the schistosomula were pre-incubated in puromycin or actinomycin D. Transient cell adherence to LS may comprise an important immune evasion stratagem by schistosomula.

Three morphologically distinct types of muscle fibres isolated from the platyhelminth Schistosoma mansoni have been studied with whole cell current- and voltage-clamp techniques. Fibres showed a marked time-dependent decrease in membrane resistance in response to depolarizing current injections. Voltage-clamp experiments revealed the presence of two distinct voltage-gated outward currents. The most prominent current is a slowly activating, slowly and incompletely inactivating potassium current similar to delayed rectifier currents which have been described in a variety of cell types from a variety of organisms. Also present is a faster activating, quickly and completely inactivating potassium current that shares functional characteristics with ‘A’-currents. All three of the cell types studied possess a delayed rectifier current, but only two of the three types have ‘A’-currents. Though depolarization with high K+ leads to contraction of the dispersed fibres, no voltage-gated inward currents could be detected by whole cell voltage-clamp under any of our conditions.

The epidemiology of trypanosome infections of Glossina pallidipes was studied at a riverine site in the Zambezi Valley, Zimbabwe for a period of 13 months. Over 9000 flies were captured using a single trap. These flies were dissected, screened for trypanosome infection, sexed, and aged using both wing fray and (for females) ovarian category indices. Midgut infections were identified to species using recently developed DNA probes. The overall prevalence of mature infections was 5·5%, comprising 3·1% Trypanosoma vivax-type and 2·4% T. congolense-type (which included very low prevalences of T. brucei, T. simiae and another Nannomonas species). The prevalence of infection increased with age. For T. vivax-type infections in flies aged by ovarian category this relationship could be described by a simple ‘catalytic’ model assuming a constant per capita rate of infection. For T. congolense-type infections this model tended to over-estimate prevalence in older age classes, implying that the rate of infection decreases with age, and/or that infected flies have higher mortality rates, and/or that a significant fraction of the population is resistant to infection. Prevalences of infection also varied between months. This variation was more marked for T. vivax-type infections and was negatively correlated with both temperature and rainfall. The shape of the age-prevalence relationship, however, did not vary significantly between months. These observations are not fully explained by variation in the age-structure of the tsetse population and are consistent with temporal variation in the rate of infection (rather than in the trypanosome developmental period or in effects of infection on fly mortality). Possible causes of this variation are discussed.

The excreted/secreted proteinases of adult and juvenile Fasciola hepatica maintained in vitro were found to hydrolyse the fluorogenic substrates Cbz-Phe-Arg- and Cbz-Arg-Arg-NHMec. This activity was demonstrated to have a classical cysteine proteinase inhibitor profile, with turn-over of both substrates being blocked by pre-incubation with E64 and peptidyl diazomethanes. The Cbz-Arg-Arg-NHMec hydrolysing activity of the mature fluke exhibited an alkaline stability not characteristic of its mammalian lysosomal counterparts. Further, the biotinylated affinity reagents biotin-Phe-Ala- CHN2 and biotin-Phe-Cys(SBzyl)-CHN2 were used to label and characterize these cysteine proteinases in terms of apparent molecular weight and subsite specificity. Adult fluke media were found to contain four species of molecular weights 66, 58, 50 and 25–26 kDa; juvenile media contained three species of molecular weights 66, 54 and 25–26 kDa. The major 25–26 kDa cysteine proteinase common to both stages was shown to have a subsite specificity similar to that of mammalian cathepsin B.

Establishment, survival and distribution of Leptorhynchoides thecatus (Acanthocephala) were investigated in largemouth bass, Micropterus salmoides, fed 10, 25, or 40 cystacanths and examined at 1, 3 or 5 weeks post-infection. Worms established widely in the alimentary tracts of bass but by 5 weeks post-infection had localized in the pyloric caeca and intercaecal region. Other individuals moved to parenteral sites where they remained immature, though viable. In the 10- and 25-level exposures, establishment and survivorship in the alimentary tract were roughly proportional to the dose of cystacanths. After 1 week post-infection in the 40-level exposure class, numbers of worms in the alimentary tract decreased significantly and parenteral occurrence increased significantly. Total survival of L. thecatus appeared to be density-independent. Maturation of worms was retarded temporarily as intensity of infection increased, but by 5 weeks post-infection worms from all doses were at roughly the same stage of development within sex. The caeca and intercaecal area apparently did not differ in their suitability for maturation.

The infection dynamics of Cyclops vicinus by the larvae of Bothriocephalus claviceps were studied experimentally. The mean number of procercoids/copepod rose to a plateau of approximately 14 as the density of eggs increased, whilst the number of copepods surviving decreased correspondingly. Copepods exposed to eggs of B. claviceps survived less well than controls, and those exposed to a higher density of eggs survived less well than those to a lower density. The mean number of parasites/copepod and the mortality of copepods were found to increase with the increasing time of exposure to eggs even when egg density was constant. These two parameters were also found to differ between gravid female copepods and unsexed young copepods. Young copepods suffered a higher parasite burden and thus a higher mortality, indicating that there are age or developmental-stage related differences in susceptibility of copepods to infection.

Nine vervet monkeys and nine baboons were infected with eggs of Echinococcus granulosus per as. Six of the vervets and one of the baboons possessed hydatid cysts at autopsy, 15–28 months post-infection. The sequential IgG response to hydatid fluid and protoscolex antigens showed considerable inter-animal variation. Infected vervets and baboons became seropositive after an average of 8 months post-infection. Considerable fluctuation in the IgG response was observed, particularly to the hydatid fluid antigen which, in humans, may contribute to the existence of a significant proportion of seronegative individuals. Vervets, in particular, may be useful to study immunological events associated with exposure, development and resolution of hydatid disease in outbred human populations.

The helminth community in the digestive tract of the sedentary predatory fish, Sciaena umbra, near the Scandola Nature Reserve, Corsica, consists of 5 species of digeneans. Anoiktostoma coronatum (in the rectum) and Metadena pauli (in the intestine) were found in almost all the fish, and in high numbers; they are considered core species. Distomum aloysiae, Pleorchis polyorchis (both in the intestine), and Stephanostomum bicoronatum (in the rectum) were relatively uncommon, usually found in small numbers, and are considered satellite species. The two core species were very abundant in June, when the size of their populations was strongly correlated with the size of the host fish. At this time, M. pauli was found mainly in the duodenum, but in high populations was spread throughout the intestine (s. strict.). Both species were much less abundant in October, their populations were less dependent on host size, and M. pauli was more limited to the anterior intestine. Pleorchis polyorchis and Distomum aloysiae were more frequent in October, were found predominately in fish with few M. pauli, and were found posterior to M. pauli, suggesting some interactions between these species. Stephano-stomum bicoronatum showed no seasonal patterns, and no apparent interactions with A. coronatum.

Density-dependent constraints on parasite growth, survival or reproduction are thought to be important in preventing the unchecked increase in parasite numbers within individual hosts or host populations. While it is important to know where, and with what severity, density dependence is acting within the parasite life-cycle, interpretation of data from natural infections is difficult. In this paper, we present a Monte Carlo simulation technique for examining such data for evidence of density dependence. We also describe how this technique may be used to distinguish among mechanisms hypothesized to generate density-dependent phenomena.

Parasitology, 105, 177–182

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The picture used for Fig. 2 represents isoelectrofocusing of B. divergens lysates.

The picture used for Fig. 3 represents isoelectrofocusing of B. hylomysci lysates.