Light and transmission electron microscopy were used to study different stages of Perkinsus atlanticus (Apicomplexa) during induced zoosporulation, with fluid thioglycollate medium and seawater. Cytokinesis and nucleokinesis of different developmental stages were studied every 12 h during the incubation period of 72 h, at which time the zoospores became free. Uninucleated and flagellated zoospores present the apical complex formed by conoid, polar ring, micronemes, rhoptries and subpellicular microtubules observed at different sections. Ultrastructural details were compared with the other two species of the genus Perkinsus.

Using cloned lines of Plasmodium berghei producing mixed asexual and sexual (clone 234L) and purely asexual (clone 233L) parasitaemias, the courses of parasitaemia, gametocytogenesis, exflagellation, ookinete production in vitro and mosquito infectivity have been followed. For clone 234L mosquito infectivity is maximal at day 3 and has ceased by day 6 post-infection. Conversely, gametocytogenesis, exflagellation and ookinete production are at minimal levels at day 3 and rise to peaks between days 10 and 15 of infection (in TO mice infected with blood at mechanical blood passage 3). Sexual potential declines progressively with sustained mechanical passage (up to P14). Gametocyte conversion is highest early in infection and declines exponentially; however, upon each mechanical passage conversion is again raised but decreases more rapidly with succeeding passages. For practical mosquito transmission in the laboratory we consider P8 to be the useful limit for mechanical transmission for this parasite clone. Asexual parasite growth (virulence) is more rapid with increased mechanical passage inducing a rapid fall in haematocrit. By P14 the course of infection closely parallels that of the purely asexual clone 233L.

Malaria and filaria infection rates were determined for anopheline mosquitoes collected whilst biting and resting in village houses in Papua New Guinea. The number of anophelines infected with both parasites was greater than expected from the infection rates of each parasite and this difference was significant in resting collections. The excess of multiply infected mosquitoes is probably a result of a vector population composed of individuals with differing numbers of opportunities to become infected. Malaria-positive, Anopheles punctulatus from resting catches had a significantly greater number of Stage 3 Wuchereria bancroftilarvae than malaria-negative mosquitoes. However, multiply infected mosquitoes appear to suffer greater mortality than non-infected or singly infected mosquitoes when the filarial worm reaches the third stage. Any potential increase in transmission resulting from multiple infections is thereby offset by a greater mortality rate in these mosquitoes.

Subtherapeutic doses of chloroquine have been reported to enhance infectivity of drug-resistant Plasmodium species to their vectors. In this investigation, Plasmodium yoelii nigeriensis N67 strain showed enhanced infectivity to mosquitoes when stimulated by chloroquine. Both sensitive and resistant clones were derived from the N67 strain by dilution, showing that this strain is polymorphic for the resistant trait. A resistant subline was derived by selection under drug pressure from a chloroquine-sensitive clone, but neither the sensitive nor the resistant clones or sublines showed enhancement of infectivity in the presence of chloroquine. This suggests that the enhancement of infectivity shown by the N67 strain is a response to chloroquine stimulation shown only by certain of the genotypes within it, and that this response to chloroquine is not a trait causally connected with the genes coding for chloroquine resistance.

An oligonucleotide mixture based on the 21 base pair repeat sequence of Plasmodium falciparum was covalently coupled to biotin and used as a probe to detect P. falciparum DNA. The limit of detection was 10 ng. This method was further developed as a fingerprint assay for parasite strain typing. After restrictian enzyme digestion, blotting and hybridization, distinct banding patterns were obtained for the strains tested and these were reproducible. In addition, discrete differences were found between PLF-3 S + /S-strains which may implicate genetic reorganization in the switching mechanism which occurs when parasites are passed from an intact to a splenectomized animal.

Brain cysts from three strains of Toxoplasma gondii were purified and analysed by SDS-PAGE and immunoblotting. Western blot analysis revealed the presence of major cyst/bradyzoite antigens at Mr 25000, 36000 and 67000 common to all strains and recognized by many individual sera. There was also evidence of strain variation in cyst antigens with, for example, antigens at Mr 88000, 28000 and 26000 unique to the 17025 strain. There was some overlap in the molecular weight of cyst and tachyzoite antigens, but the major tachyzoite surface antigen (P30) could not be detected in brain cysts.

Cell cloning techniques and schizodeme analysis were used to detect mixtures of subpopulations in Leishmania parasites, isolated from humans and a reservoir host. Clones were obtained by plating promastigotes, at limited dilution, on solid medium. The resultant colonies were analysed by the restriction profile of kinetoplast DNA minicircles (schizodeme analysis). The efficiency of plating was around 40–60%, and subpopulations of different schizodemes, in stocks of Leishmania isolated from 2 cases of human cutaneous leishmaniasis were detected. The presence of different schizodemes in one isolate from a sylvatic animal suggested the possibility of a mixed natural infection.

Reinfection pattern among 6- to 20-year-old subjects was studied over 24 months in two Mauritanian villages of intenseSchistosoma haematobium infection after a targeted chemotherapy with praziquantel involving the whole community. Subjects received treatment according to the presence of haematuria/proteinuria and this indirect screening technique was able to identify 98–100% of the heavily infected subjects (50 + eggs/10 ml). The two villages differed with respect to their characteristics, quality of follow-up and reinfection pattern. The post-treatment 6-month cumulative incidence during the two transmission periods following the chemotherapy, estimated from a subset of 116 subjects, was 18·0% and 20·5%. Reinfection rates were higher among males (Cox-Mantel: P = 0·0015), among children 6–10 years of age than older (P = 0·0078) and among subjects with more than 50 eggs/10 ml of urine before treatment than subjects with a lower egg output (P = 0·009). A Cox proportional hazard regression model was fitted and confirmed that gender, age and pretreatment level of infection were predictors of the rate of reinfection but that there was no interaction between these predictors.

The toxicity of the active oxygen species hydrogen peroxide, superoxide radical, hydroxyl radical and singlet oxygen to microfilariae (mf) has been studied in vitro, using active oxygen-generating systems and scavengers/inhibitors. Mf viability was monitored by uptake of the radiolabel, [3H]2-deoxy-D-glouse. Hydrogen peroxide and singlet oxygen, but not superoxide radical or hydroxyl radical, are toxic for mf. Hydrogen peroxide was toxic for mf within 2 h at concentrations as low as 5 ¼, an amount eosinophils have been shown to release in vitro (Weiss et al. 1986). Catalase and thiourea, but not inactivated catalase, superoxide dismutase (SOD), singlet oxygen scavengers, or hydroxyl radical scavengers, protected mf. Mf have relatively high levels of endogenous SOD but no measurable glutathione peroxidase and low levels of catalase when compared with other parasites (Callahan, Crouch & James, 1988). The low levels of hydrogen peroxide-scavenging enzymes correlate well with mf sensitivity to hydrogen peroxide and the protective effect of exogenous catalase.

The short-lived infective copepodid stages of the copepod Salmincola edwardsii, ectoparasitic on brook trout, Salvelinus fontinalis, are under strong selective pressure to evolve efficient host-finding mechanisms. In laboratory experiments, we quantified the effects of visual and mechanical stimulation on the activity of the copepodids. We found that shadows passing above the copepodids and shock waves passing through the water generated marked increases in the rate and length of upward swimming bursts, which resulted in more time spent suspended in the water column and greater distances travelled. However, we found no strong evidence for a relationship between the magnitude of the copepodids' response and the intensity of stimulation. Although these responses are not host-specific, they allow the parasites to conserve their limited energy stores by increasing their host-finding activity only when potential hosts are nearby.

Infectivity of Pseudechinoparyphium echinatum cercariae to 11 species of gastropod was examined experimentally. Broad specificity and differential host—parasite compatibility were exhibited. Nine gastropod species functioned as second intermediate hosts. Planorbarius corneus, Physa fontinalis, Lymnaea peregra and Biomphalaria alexandrina showed high levels of compatibility with the parasite. In single-species exposures over 90% of cercariae encysted in each of these hosts. Low compatibility with the first intermediate host species Lymnaea stagnalis may be a mechanism preventing super-infection of emitting snails. Cercariae did not infect the prosobranchs Bithynia tentaculata and Viviparus viviparus. Experimental infection of a host community comprised of 8 European gastropod species revealed an order of host utilization similar to that shown in single-species exposures. However, cercarial transmission success in P. fontinalis and L. peregra (compared to that in P. corneus) was significantly reduced. This may have been due to the marked preference to cercariae for P. corneus compared to the other two highly suitable hosts for whom cercariae showed equal preference.

We previously reported that blood forms of Trypanosoma cruzi express alpha- and beta-adrenergic receptors and that binding of specific agonists to these receptors modifies the infective capacity of the parasite in vitro. The present study has revealed that the inhibitory effect of the beta-adrenergic agonist L-isoproterenol and the stimulatory effect of the alpha-adrenergic agonist L-phenylephrine are not produced when the parasite is subjected to prolonged exposure to otherwise effective doses of these agonists or when supraoptimal doses of these agonists are used. We refer to these phenomena as ‘desensitization’ because of their analogy with vertebrate cells becoming desensitized by prolonged exposure to, or relatively high concentrations of, adrenergic agonists. At a constant agonist concentration, T. cruzi desensitization was time-dependent and, when the time of parasite treatment with the agonists was not changed, the higher concentrations of the agonist tested were the most effective in producing desensitization. The reduced infectivity resulting from treatment with optimal doses of L-isoproterenol was accompanied by elevated levels of cyclic adenosine mono- phosphate (cAMP) which were not detectable when L-isoproterenol concentrations producing desensitization were used. This finding implicated cAMP as a likely second signal in the inhibitory mechanisms of this agonist. No significant change in cAMP was detectable in parasites treated with L-phenylephrine, leaving open the question about how optimal doses of this alpha-adrenergic agonist enhance T. cruzi infectivity. Parasite responsiveness to alpha- and beta-adrenergic agonists as well as the desensitization effects define a system which regulates infectivity and could be modified at the host tissue level by naturally occurring agonists.

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.

Metacestodes of Hymenolepis diminuta cause a perturbance of vitellogenesis in the intermediate host Tenebrio molitor. The reduction in host reproductive output associated with infection may be due to this pathophysiology. Many of these events are regulated by host juvenile hormone (JH). A comparison of the titre of JH and its rate of degradation in female control and parasitized 15-day-old insects has been made. Haemolymph from female beetles contained 1·27 pMol JH equivalents/100 µl. No significant difference was associated with infection. However, topical application of a JH analogue, methoprene, at the time of infecion or 8 days post-infection reduced the significant accumulation of vitellogenin usually found in the haemolymph of females 12 days or more post-infection. These findings indicate that parasite-induced alteration of host vitellogenesis is not mediated via alteration in JH titres, although observations made after hormone supplementation suggest some form of interaction between the parasite and the host endocrine system.

Vesiculated protoscoleces (VP) were produced by culturing freshly collected protoscoleces from Echinococcus granulosus horse liver hydatids in RPMI 1640 monophasic medium at 37°C for 18 days. Half of the VP were used as such, the other half used after killing them by freeze—thawing. Nine-day-old chicken heart fragments (CHF) were cultured in MEM at 37°C for 72 h. Subsequently, CHF were put together with live and dead VP, respectively, for up to 53 days, on a semisolid medium consisting of agar, Ringer's and MEM. Time-dependent histological observations revealed that dead VP were surrounded by CHF cells. Dead VP tissue was eventually internalized and disintegrated in about 1 week. Live VP penetrated into the CHF tissue and further developed into small hydatid cysts, located within the boundaries of the experimental ‘host’ tissue. The amorphous-looking contact region PAP-stained positively only with anti-E. granulosus serum and not with anti-CHF serum; it was considered identical to the normal laminated layer. The invasion of VP in CHF tissue proved to be different from a tumour or a bacterial invasion: it was concluded that the confrontation of VP and CHF had resulted in an ‘in vitro cohabitation’ rather than in an ‘in vitro infecion’.

A size selected genomic DNA library was constructed using DNA extracted from Taenia saginata. The DNA was digested using the restriction enzyme EcoR1 under star conditions and the 2–4 kbase fraction, selected following sucrose density-gradient separation, was cloned in the bacteriophage λgt 10. A panel of cestode DNAs including Taenia saginata, Taenia solium, Taenia taeniaeformis, Taenia crassiceps, Echinococcus granulosus and DNAs of bovine, porcine and human origin were used in conjunction with hybridization analysis to identify two recombinant bacteriophages. The first probe, designated HDP1, reacted specifically with T. saginata DNA. The second, designated HDP2, reacted with DNA from both T. saginata and T. solium but not the other DNA samples and thus provided a general reagent for positive identification of fragments of Taenia spp. proglottides of human faecal origin. If used in conjunction the two DNA probes allow positive identification of T. saginata. In the clinical situation it is important to be able to distinguish T. saginata and T. solium infections and DNA probes such as these may be useful in such differentiation.

Taenia saginata cyst fluid proteins from 4, 8, 12 and 16-week-old cysticerci were analysed by a combination of direct I radio-isotope labelling, immunoprecipitation using a panel of sera from infected cattle infected with T. saginata and SDS–PAGE. Protein antigens of 12, 14, 16, 20 and 26 kDa were identified in all of the cyst fluids examined. These were immunogenic and were precipitated by serum taken from cattle from 8 weeks after infection onwards and were therefore considered to be of diagnostic potential. A 185 kDa protein antigen found only in the cyst fluid of 4-week-old cysticerci and a 43 kDa protein antigen first detected in cyst fluid from 8-week-old cysticerci were also identified but were considered to be of more limited diagnostic potential due to their restricted presence. An apparently non-immunogenic 67 kDa protein, found in all the cyst fluids examined, may have been host serum albumin.

Neonatal hamsters were exposed to varying doses of Necator americanus larvae and changes in the stability of the resulting worm burdens were monitored over a period of 25 weeks. No change in worm burdens was evident for the first 5 weeks of infection, irrespective of the infection intensity, but the more heavily infected groups subsequently lost worms in a density-dependent manner. Male and female hamsters lost comparable proportions of their established parasite burdens indicating that there was no host sex-linked difference in this respect. By week 15 infections had stabilized and the residual worm burdens, usually a maximum of 30 worms survived for a considerably longer period of time. Initially the percentage of male worms varied from 45% to 50% but as infection progressed male worms comprised a significantly increasing proportion of the total worm burden. By week 25 the percentage of male worms was usually in excess of 60%. The growth of infected animals was not severely affected by N. americanus, even when heavy worm burdens established initially, but a significant effect was detected particularly in week 5, prior to worm loss, when the adult worms would have been feeding on intestinal tissues and causing blood loss for a period of about 2 weeks. The most severe depression in the packed cell volume was also recorded in week 5, indicating that anaemia had been initiated in infected hamsters. Whilst, the regulation of parasite burdens in weeks 5–10 post-infection may have resulted from host immunity, the persistence of the residual worm burdens, the marked density-dependent anaemia and the subtle effect on host weight, all reflected well-documented aspects of chronic human necatoriasis.

Strains of mice poorly (B10) or non-responsive (B10.BR) to a primary infection with Trichuris muris were protected against infection by vaccination with excretory/secretory (E/S) antigen in Complete Freund's Adjuvant (CFA). Protection in these mice was slow to be expressed compared to that in good responder strains. Vaccination boosted the IgG and IgG1 antibody responses to E/S antigen and altered the antigen recognition profiles, three high molecular weight antigens (80–85, 90–95, 105–110 kDa) being recognized by antibodies in sera from vaccinated but not control mice. B10. BR mice which had experienced a patent primary infection could not be protected against challenge infections by vaccination and this was correlated with depressed levels of IgG1, but not total IgG, to E'S antigen early post-challenge compared with vaccinated infected mice which had not seen an adult primary infection. There was also lack of recognition of the three high molecular weight antigens recognized by antibodies in sera from mice infected after vaccination. It is suggested that the rapid development of high levels of IgG1 antibodies, and the recognition of the three high molecular weight antigens, may reflect events that are important in protective immunity. Immunomodulation of host immunity by T. muris may therefore be achieved, at least in part, by the suppression of specific IgG1 levels, the production of an irrelevant IgG isotype and prevention of the recognition of critical antigens.

Comparisons were made of the infectivity and antigenicity of 4 Trichinella spiralis isolates (S, D, Y, W), of quite different geographical origins, and T. pseudospiralis (P) in rapid- and slow-responder inbred mice. Infectivity was measured by the Index of Reproductive Capacity (ICR) expressed as the ratio between the number of muscle larvae recovered on day 30 post-infection (p.i.) and the numbers of larvae given at infection. Antigen recognition was measured by the degree of proliferation of mesenteric lymph node cells (MLNC) to in vitro stimulation with crude muscle larvae antigen (CMLA) and by the total antibody responses to CMLA at day 25 p.i. as measured by ELISA. Regarding infectivity the isolates fell into two groups, high infectivity (S, D and Y) and low infectivity (W and P). Analysis of CMLA, detergent-stripped (CTAB) and I-labelled surface larval proteins was made by SDS—PAGE under reducing conditions. Differences in antigen profiles were seen in all antigen preparations, being most noticeable in CTAB and 125I-labelled proteins from W and P isolates. Antigen recognition by polyclonal infection-derived antisera and by monoclonal antibodies raised against the T. spiralis London strain (L) was studied in the W (Arctic) and S (Spanish) isolates. Polyclonal antisera recognized different antigens in the S and W isolates, as did the monoclonal antibody, although recognition was more restricted. Neither antibody recognized a 64 kDa band in the W isolate which was clearly visible in the others tested.