We examined a host system (Peromyscus maniculatus) in which a single autosomal gene controls susceptibility or resistance to infection by the cestode parasite Hymenolepis citelli. Parasite deaths of both primary and secondary (challenge) infections were examined, using a probabilistic model, to see if deaths were random and uncorrelated within each genotype. Within susceptible hosts, post-patent survival of primary worms was not random and heterogeneity was due to among-host effects rather than parasite-age effects, suggesting a second genetic or immunological process. Secondary infections in susceptible hosts appear to be lost randomly, independent of primary infection age or burden. The loss rate is similar to that for primary worms. The lack of correlation or alteration in (susceptible) host response to primary and secondary infections suggests that the latter are eliminated by a process that differs from that acting on primary worms. In resistant hosts, parasite survival rates suggest that the immunological process elicited by the primary infection also acts to eliminate the secondary infection more rapidly. Suggestions are made for improving experimental methods when dealing with mixed genotype populations. Experiments should permit direct estimation of genotype frequency and of parasite death rates within each genotype. This separation of host type is particularly important when studying correlation of successive worm burdens since any host or treatment mixture (genotype, age, sex, size or infection dose) may induce correlations that could be mistakenly interpreted.

This study describes experiments which demonstrate the presence of a parasite-induced or derived factor in the haemocoel of Onchocerca lienalis-infected simuliids. Haemolymph, when transferred from previously infected donor flies to previously untreated recipient flies, using fine glass needles, confers protection against a subsequent challenge to the recipients. In three trials using Simulium ornatum sl. or S. linealum, significant levels of protection in ‘immunized’ compared to ‘challenge control’ groups were achieved (84% and 81% with S. ornatum sl. and 85% with S. lineaturm). This is despite the fact that the worm burdens of the infected donor groups in the three trials were very different (means of 33·1 and 3·4 larvae/fly with S. ornatum sl. and 1·6 larvae/fly with S. linealum). The use of sham-operated and bentonite-inoculated control groups demonstrate that this effect is not merely a non-specific reaction to trauma, but is probably parasitic in its derivation. The use of double infections separated by 4 days shows that the donor flies have themselves probably acquired resistance to O. lienalis. Frequency distributions of O. lienalis burdens in individual flies show that there is a shift froni a normal to a skew distribution when comparing ‘challenge control’ flies with ‘immunized’. This possible acquired resistance could have important implications in the transmission of forest and savanna human onchocerciasis by the Simulium damnosum complex.

Eimerian oocysts were found in 98% of 422 faeces samples from goats in SE England. It was found that 74% of the samples contained thousands or tens of thousands of oocysts/g. Kids less than 1 year old had higher oocyst counts than yearling or adult goats. Nine species of Eimeria were identified, and 65% of the samples contained 3–5 species. The most prevalent species were E. arloingi (94%), E. hirci (69%), E. christenseni (64%) and E. caprina (55%). E. ninakohlyakimovae (48%), E. alijevi (42%) and E. apsheronica (23%) were common, and E. joichijevi (8%) and E. caprovina (4%) were comparatively rare. Developmental stages of E. caprina destroyed the crypt epithelium in the mucosa of the large intestine, causing severe haemorrhagic enteritis similar to that produced by E. ninakohlyakimovae. The literature on Eimeria species in sheep and goats is reviewed and, on the grounds of priority, E. bakuensis is the correct name for E. ovina in sheep.

In merozoites of Plasmodium knowlesi, rhoptries have a dense substructure of fine (2·5 nm diameter) granules and short rods. These are not altered by lipid extraction, and stain with ethanolic phosphotungstate indicating a proteinaceous composition. Various types of fixation also show multilamellar whorls with a periodicity of 5–7 rim in the tips of rhoptries or extruded at the merozoite apex. In merozoites fixed during invasions of red cells, membrane continuity typically occurs between the rim of the rhoptry canal and the red cell membrane, but where this contact has apparently been lost, extensive membranous whorls and blebs are often found at the apex of the parasite. Similar structures occur at the spices of merozoites within late-stage schizonts. It is suggested that the same mechanism which generates these lamellae forms the parasitophorous vacuole by inserting membranous elements formed by the parasite into the red cell membrane, so causing its invagination. A similar mechanism may be responsible for the release of merozoites from the late-stage schizont.

Electrophoresis of extracts of schizonts of Plasmodium knowlesi in non-dissociating polyacrylamide gels, separates several bands of acid endopeptidase activity. A polyclonal antiserum, produced by immunization with purified merozoites, failed to distinguish between different bands of the parasite enzyme, indicating that they are serologically related. Apart from the loss of one minor peak, extraction in Triton X-100 did not reduce the enzyme's electrophoretic heterogeneity. The antiserum did not react with red cell acid proteases.

Polyclonal antisera raised against Plasmodium knowlesi reacted with (1) NADP-specific glutamate dehydrogenase (GLDH) of P. knowlesi, (2) GLDH of P. falciparum and (3) GLDH of Proteus spp. The antisera did not react with NAD(P) GLDH from bovine liver. Polyclonal antisera raised against the GLDH of Proteus spp. cross-reacted with GLDH from P. falciparum. Monoclonal antibodies (McAbs) obtained from mice immunized with Proteus GLDH were either specific for the bacterial enzyme or cross-reacted with P. falciparum GLDH. The selected McAbs did not react with GLDI-1 from P. knowlesi, P. chabaudi or P. berghei. The GLDH of P. falciparum was shown to be a cytosolic protein (by FAT) with a subunit molecular weight of approximately 49000 Da (by immunoprecipitation) having a pre dominantly hexameric form (by sucrose density gradient). Implications of the conserved sequences of GLDHs and other enzymes are discussed.

The induction of schistosomulicidal activity of peritoneal macrophages by concanavalin A-stimulated supernatants from long-term T-cell clones and by interferon-gamma (IFN-γ) was investigated in detail. Optimal conditions of in vitro macrophage activation by T-cell clone supernatants were established. Macrophages from 13-week S. mansoni-infected mice responded to lymphokine activation as well as resident mnacrophages from uninfecteci mice. IFN-γ was shown to play an essential role in induction of schistosomulicidal macrophage activity: recombinant IFN-γ at high concentration could induce schistosomula killing, and an anti-IFN-γ antiserum inhibited the induction ofschistosomulicidal activity by T-cell clone supernatants. Our data also indicate that macrophage activation could be obtained by IFN-γ in synergy with other lymphokines in the supernatant of long-term T-cell clones. Macrophages from mice injected with T-cell clone supernatants were primed in vivo and triggered to kill schistosomula in vitro in the presence of lipopolysaccharide (LPS). The data demonstrate that lymphokines produced by T-cell clones and, in particular, IFN-γ can participate in the activation of schistosomulicidal macrophages.

Daughter sporocysts (SpII) of Schistosoma bovis produced by the mother sporocyst (SpI) can direct their production along three different pathways during intramolluscal development. (i) Direct sporocystogenesis–tertiary sporocysts (SpIII) are directly produced by non-cercariae producing daugher sporocysts (SpIII). (ii) Simultaneous sporocystogenesis and cercariogenesis–SpIII are produced by Spil in which cercariae are being produced. (iii) Sporocystogenesis after cercariogenesis-Spill are produced by SpIl in which cercariae are undergoing degeneration. These mechanisms can be interpreted in terms of demographic strategy leading swiftly to the taking over and optimum exploitation of the host's hiotic capacity.

The migration of schistosomula through the lungs of mice has been investigated at the ultrastructural level. On Days 4 and 7 post-infection schistosomula were located only within the vasculature. Newly arrived Day 4 schistosomula were highly convoluted and completely occluded vessels measuring approximately 15–25μm in diameter. By Day 7 parasites were located in pulmonary capillaries, often causing considerable distension. The subtegumentary fibrous interstitial layer was absent in elongated lung schistosomula. The minimum diameter of such parasites was 8μm, and their tegument was usually unridged and flattened against the vascular endothelium, thus resulting in close apposition of endothelial and tegumental membranes. The tight fit of schistosomula in the pulmonary capillaries implies that migration through them is both a strenuous and slow process. The migratory delay in the lungs after arrival from the skin occurs because dimensional changes are necessary for successful capillary migration.

Previously quoted figures for the frequency of antigen switching in Trypanosoma brucei are based on incorrect assumptions. In order to determine the correct switching frequency, an equation was derived that takes the growth rates of the newly expressed antigen types into consideration as well as the proportion of switched trypanosomes and the number of generations since the population was antigenically homogeneous. When this equation was applied to published in vitro data, variable values were obtained for the switching frequency in clonal populations originally expressing one antigen type. The calculated most likely switching frequencies ranged from 1·4×10−7 to 3·5×10−6. This variation was probably caused by differences in the growth rates of the new antigen types in the population and failure to detect slow growing variants. To overcome these problems, an experimental procedure was developed to analyse the switching frequency in vitro. Trypanosomes were cloned and grown in parallel cultures. After an appropriate number of generations, cells expressing the original antigen type were destroyed and, from the proportion of cultures that contained new antigen types, the switching frequency was calculated. The technique minimized subculturing or other procedures that could distort the results. Although the method was optimized for analysing switching frequency, the values differed between experiments, ranging from 2·2×10−7 to 2·6×10−6 for one variant. Possible causes for the variations in switching frequency are discussed.

The course of the migration of infective larvae of Brugiapahangi within the mosquito, Aedes aegypti, is described. The presence of blood or eggs in the abdomens of mosquitoes at a time when the infective larvae are beginning their migration out of the thorax leads to a significant increase in the proportion of larvae accumulating in the abdomen. The evidence supports the hypothesis that infective larvae may accumulate in the labium as a direct consequence of the insect's internal anatomy i.e. infective larvae on moving into a confined space tend to remain there. In addition, the rate at which infective larvae migrate into the labium is shown to be density dependent.

A variety of morphological forms developed during monophasic culture of ruptured brood capsules of equine Echinococcus granulosus. Attached to some protoscoleces were small posterior bladders, with a surface ultrastructure of truncated microtriches, and these never developed a laminated layer. In contrast, small free vesicles of similar size and ultrastructure, but devoid of an attached protoscolex, produced a layer by 20–30 days. The surface ultrastructure of both these forms is similar to that of the protoscolex attachment stalk and the brood capsule lining, and posterior bladders and small free vesicles are considered to be derived from portions of the stalk±part of the brood capsule lining retained at protoscolex release. Also present were large vesicles interpreted from their size and surface anatomy as ruptured, everted brood capsules. When devoid of attached protosoleces these formed cysts each with a laminated layer, whereas similar vesicles with several attached protoscoleces did not develop a layer, or only very occasionally after 70 or more days. In contrast, a few large vesicles with a single attached protoscolex produced a laminated layer. The possibility that protoscolex attachment to stalk remnants or brood capsules inhibits the cystic development of these structures is discussed.

The reaction of Balb/c mice to primary and secondary subcutaneous infection with chinoeoccu.s granulosus protoscoleces (PSC) is described. From 3 to 14 days following primary exposure to PSC, draining lymph nodes increase in weight and there is expansion of T and B lymphocyte populations, enhancement of in vitro lymphocyte transformational responses and production of PSC-specific 1gM and IgE antibodies. Despite the persistence of viable PSC in host tissues, lymphocyte responses decline to pre-infection values over the period 3–8 weeks post-infection. Secondary exposure to PSC immediately induces lympho proliferation, enhancement of transformational responses, production of IgE antibody and encapsulation of PSC by inflammatory cells. Although specific antibody levels remain high until at least 8 weeks after challenge infection, lymphocyte activity begins to decline after 4 days and is profoundly suppressed by 10 days. Parasite viability appears to be significantly reduced in secondary, as opposed to primary, infection and is associated with the accumulation of large numbers of eosinophils, mast cells and macrophages in infected tissues.

Some effects of Hymenolepis diminuta on the fecundity of Tribolium confusum are described. Host fecundity is observed to be reduced exponentially with increasing parasite burden/host, although there are differences in the ability of individual hosts to respond to parasitism. Of particular interest is the finding that host fecundity is greatly reduced in young beetles on or by day 14 post-infection (p.i.). This age-related reduction in host fecundity is discussed in relation to the population dynamics of this hostr-parasite relationship.

The role of serum in the adherence of roach (Rutilus rutilus) leucocytes to the plerocercoid of Ligula intestinalis has been investigated in vitro. Roach plerocercoids, either untreated, cultured or killed (fixation in 0·5 % buffered glutaraldehyde or heat at 50°C for 45 mm), were exposed to serum and 4×106 leucocytes obtained from the pronephros of non-infected roach using a discontinuous Percoll gradient. At normal roach serum (NRoS) dilutions of 1:5000 to 1:100, leucocyte adherence was observed on all living parasites but was negligible in killed parasites or in complement-depleted NRoS assays. The number of adherent cells increased with increasing NRoS concentration. Leucocytes bound more avidly in the presence of heat-inactivated immune serum (hi IRoS); at a serum dilution of 1:5000 the percentage of the parasite surface covered by leucocytes was approximately 3 times greater in hi IRoS than in NRoS. Ultrastructural observations on parasites revealed that plerocercoids exposed to cells and hi IRoS had, in some areas, abnormal inclusions in their tegument and lacked a microthrix border and distal cytoplasm. In addition, the efflux of radioactivity from [14C] cycloleucine-labelled worms was greater in parasites incubated in the presence of hi IRos.

Interpretation of anthelmintic activity using in vitro screens has, until now, relied on the detection of drug-induced effects on nematode development, viability and motility. A novel biochemical parameter dependent upon the spectrophotometric assay of acetyicholinesterase (AChE), an enzyme secreted in large quantities by certain trichostrongylid nematodes, has been developed to replace these often subjective indices of activity. Using Nippostrongylus brasiliensis, a worm frequently employed for primary screening, the secretion of this enzyme in the presence or absence of a large number of drugs in vitro was determined. During a 4-day incubation period in a complex undefined medium without serum, AChE was secreted by normal 4th larval and immature adult stages of the worm in a linear fashion. All modern broad-spectrum veterinary anthelmintics, regardless of their mode of action, dramatically reduced the amount of enzyme secreted. Correlation between the biochemical and observa tional parameters was excellent and the selectivity of the assay when based solely on enzyme secretion was not lost. Other advantages were that the time required for the activity of certain slow-acting compounds to be detected was reduced from 7 to 4 days and that close microscopical examination of the worms was not necessary.

Environmental sex determination in the mermithid nematode Romanomermis culicivorax is examined in the context of parasite reproductive success and population regulation. Experimental results show that the sex ratio of the nematode within its mosquito host (Culex quinquefasciatus) is dependent on parasite density. Sex ratios are biased to females at low parasite burdens and to males at high parasite burdens. Low temperature further enhances female-biased ratios. The net effect of density-dependent sex determination on parasite and host population growth is shown to be critically dependent on the frequency distribution of parasite numbers/host. Mermithid parasite distributions within natural host populations show low degrees of aggregation relative to other helminth species. The population regulation of the parasite is examined with respect to environmental sex determination and parasite-induced host mortalities by means of simple mathematical models of the dynamics of parasite transmission via its life-cycle. The significance of regulatory constraints on population growth are discussed in relation to the use of mermithids as biological control agents of insect pests or disease vectors.

The in vitro uptake of gamma-emitting radionuclides by microfilariae of Dirofilaria immitis was investigated. Radionuclides tested were 133Ba, 207Bi, 82Br, 109Cd, 51Cr, 60Co, 59Fe, 203Hg, 125I, 54Mn, 32P, 125Sb, 75Se, and 65Zn. Only 207Bi, 59Fe, 203Hg, 54Mn, and 46Sc showed more than 2% of the available radioactivity to bind to the microfilariae. When tested for retention in vitro only 203Hg showed retention levels approaching 90%. Moreover, when dimethyl suiphoxide was incorporated into the medium at levels of 1% (v/v) the uptake of 203Hg could be increased by 3–5 times; no other radio-isotope tested responded in this manner. The uptake of 203Hg was directly related to temperature and time of incubation. Mercury, as mercuric chloride, was toxic to the microfilariae and respresents an impediment to the incorporation of high levels of 203Hg in microfilariae.

Microfilariae of Dirofliaria immitis were labelled with 203Hg 2+in vitro and injected into irradiated mice and Beagle dogs. With irradiated mice it was possible to demonstrate microfilariae present in the blood and to detect 203Hg by external counting as long as 28 days after dosing. The 203Hg 2+ label had a half-time of 4–5 days; the amount of stable mercury in the labelling medium strongly influenced the survival of microfilariae in vivo. In dogs, external counting showed the lungs to be a major location of the microfilariae soon after reinjection into the host. Evidence was obtained that labelled microfilariae can circulate; however, the detection of dispersed microfilariae is difficult because of the relative insensitivity of the detecting system. For radiomercury the accumulation of the inorganic form in the liver and kidneys limits the long-term usefulness of 203Hg 2+ as a label if the organism being studied also accumulates in these organs.

A deterministic model of the population biology of the non-infective free-living phase of Ostertagia ostertagi is described and tested. A comparison of field and laboratory studies indicates that the pre-infective larvae of O. ostertagi spend about twice as long in this phase in the field as in laboratory culture (13 days and 7 days respectively at 15 °C), and that the subsequent transition (i.e. second moult) to the infective phase also proceeds at a much slower rate (about 10 times less at 15 °C) in the field. Because of this discrepancy, estimates of model parameters are based entirely on measurements from field studies. A two-parameter model of pre-infective development in the field is derived. It describes egg and larval development as a direct non-linear function of air temperature. We also show that the mortality of the eggs and pre.infective larvae increases with temperature (although this relationship probably also subsumes a dependence on faecal moisture content). The model is shown to be a good mimic of the population dynamics of the non-infective free-living phase when tested against an independent data set, and is the basis of a larger model of the epidemiology of ostertagiasis.