Toxocara canis infective larvae are known to produce abundant glycosylated molecules which may be found associated with the surface or secreted into their environment. Using a range of fluorescein-conjugated and gold-conjugated lectins, the localization of particular carbohydrates was defined on the surface of live parasites, and internally at the ultrastructural level. Surface exposure of N-acetyl galactosamine and N-acetyl glucosamine was deduced by binding of FITC-conjugated Helix pomatia (HPA) and wheat-germ agglutinins (WGA). These sugars appear to be associated with a densely staining surface coat as conventional immuno-electron microscopy procedures dissipate this coat and reveal no surface binding site for these lectins. However, by using cryo-immuno-electron microscopical (C-IEM) techniques, the surface coat is retained and can be shown to bind WGA. The fluorescent lectins also revealed strong WGA binding to the secretory and amphidial pores, while the buccal opening and the cuticular alae bound HPA. Corresponding results were obtained at the ultrastructural level. Thus, HPA bound to the electron-dense area of the cuticle, areas of local cuticular thickening such as the alae and buccal labia, as well as to the oesophageal lumen. WGA also bound to the thickened cuticle of the alae and the buccal opening, but showed no reaction to either the electron-dense layer of the cuticle or the oesophageal lumen. Unlike HPA, WGA did bind specifically to the secretory column contents and the electron-dense regions of the lips associated with the chemosensory amphids. The compartmentalization of the sugars N-acetyl galactosamine and N-acetyl glucosamine, their sources and routes of surface expression and the possible association with the TES glycoprotein antigens are discussed.

Genetic influences upon host variation in eosinophilia and resistance to helminth infection, and the relationship between these parameters, were investigated in 7 inbred and 1 hybrid strains of mice infected with Trichinella spiralis. Clear strain-dependent variations were observed in the maximum peripheral blood, bone marrow and spleen eosinophilia attained in infected animals. SWR, NIH and SJL strains of mice all gave high responses to infection; four congenic strains sharing the B10 background (C57BL10 [B10], B10.S, B10.G and B10.BR) were low responders. Some of the genes for high responsiveness appeared to be dominant, as F1 hybrids from high- and low-response phenotype parental strains showed intermediate to high responses to infection. Intestinal eosinophilia showed no correlation with either peripheral blood or bone marrow responses (NIH and B10 strains having similar levels of eosinophil response in gut tissue) and was unrelated to the level of resistance to infection. Whereas NIH were highly resistant, with adult worm burdens at 13 days post-infection and muscle larval burdens at 35 days post-infection significantly lower than all other strains, B10 were quite susceptible, retaining substantial worm burdens at day 13 and harbouring large numbers of muscle larvae. Measurements of the level of the eosinophilopoietic cytokine IL-5 in sera during infection showed that the two strains differed in the kinetics of release but not in their absolute capacity to produce this cytokine. NIH mice released high levels during a primary infection, B10 released high levels during a secondary infection. The relationship between eosinophilia and resistance and the differences seen between responses in different body compartments are discussed in terms of genetically determined influences operating at the levels of T helper cell populations, T cell cytokines and bone-marrow precursor cell populations.

The epidemiology of nematode infections of Soay sheep on the island of St Kilda over a period of 2 years (August 1988–August 1990) spanning a host population crash is described. Infective larvae (L3) levels on pasture were high (2422±365 L3/kg D.M. grass in midsummer 1988) when host population density was high, decreasing after the sheep population declined by 70% in early 1989 (601 ±14 L3/kg D.M. in midsummer 1989). The availability of infective larvae to sheep increased during the winter of 1988–1989, probably as a result of concentration of existing larvae on grass as vegetation was destroyed by bad weather and overgrazing. Increased availability of pre-parasitic stages was accompanied by a marked increased in faecal egg counts from sheep of all ages and both sexes. Prevalence and intensity of infection (faecal egg counts) were higher in males than females throughout the 2-year study (χ2 = 208.3, P < 0.005 and F1.2000 = 304, P < 0.001 respectively), except during the lambing periods, and decreased with age in both sexes. Changes in prevalence and intensity of strongyle infections were associated with changes in host population density. Prevalence and intensity of Dictyocaulus filaria larvae in faeces increased during the host population crash. Infection intensity decreased with age (F1.203 = 44.02, P < 0.001) and was higher in males than females (F1.203 = 13.45, P < 0.001).

An analysis of interstrain variation between 12 inbred and 4 congenic rat strains in the expression of immunity against Trichinella spiralis is reported. All rat strains expressed strong rapid expulsion which resulted in the elimination of 88–98% of a challenge infection of muscle larvae. In contrast, substantial interstrain variation in the rate of adult worm expulsion in the primary infection as evident. By day 10 after infection, BUF and YO strains had < 50 worms left in the intestine whereas BI and WKA strain rats had barely begun rejection, with approximately 1000 worms present in the gut for both strains. All other rat strains fell within these extremes in a continuous gradation. There was no clustering of rat strains into phenotypic groups with comparable worm burdens as seen with mice. The number of muscle larvae that established after the primary infection showed less variation than had adult worm burden in the primary infection and there was only a weak correlation of muscle larvae burden with numbers of intestinal adults present at 10 days. Comparison of MHC matched or MHC-disparate rat strains on a PVG background suggested that non-MHC genes determined the principal adult worm rejection characteristics of a given strain. The absence of phenotypic variation in the expression of rapid expulsion in rats reinforces the biological distinction between rat rapid expulsion and the ‘rapid expulsion’ defined for mice

Toxocara canis infective stage larvae continually produce excretory–secretory (TES) glycoproteins in long-term in vitro culture. The kinetics of synthesis and secretion were studied by metabolic labelling with radioactive [35S]methionine, [14C]serine and [14C]threonine. Maximal incorporation rates required overnight pre-incubation of parasites in medium depleted of the appropriate amino acid. Larvae rapidly incorporated isotope into their somatic tissues, but there was a minimum delay of 10 h before secretion of labelled antigens. Labelling with [14C]serine and [14C]threonine demonstrated a relative abundance of these amino acids in the major surface/secreted glycoproteins of this nematode (TES-32 and 120). Pulse-chase experiments suggested that TES-120 may be derived from a 58 kDa precursor, reflecting extensive post-translational glycosylation. Inhibition of N-glycosylation with tunicamycin and digestion with N-glycanase provided evidence of N-glycosylation in the lower molecular weight ES components (TES-32, 55 and 70). These agents had no effect on the higher molecular weight components (TES-120 and 400) implying that for these molecules glycosylation is predominantly O-linked. The largest ES component (TES-400) was unusual, in incorporating serine and threonine but not methionine, and by exhibiting increased apparent molecular weight following pronase digestion; it is suggested that this molecule is a proteoglycan.

Laboratory experiments indicated that the active life-span of the cercariae of Cryptocotyle lingua was temperature-dependent. An increase in water temperature and population age both correlated with a tendency for the organisms to become decaudate. The larvae were strongly photoresponsive to lateral light but with time, horizontal swimming rates (HSRs) from release point to light source progressively declined. When measured over a range of light quantities, HSRs peaked at 30 μM/m2/s. HSRs were also influenced by water temperature. A rapid increase occurred up to 15 °C after which there was a precipitous decline. HSRs to coloured light were negatively correlated with increasing wavelength. When offered a choice between colours, cercariae favoured the shorter wavelengths. The implications of these responses for successful transmission are discussed.

In early 1989, two-thirds of the Soay sheep population on St Kilda died over 12 weeks. Post-mortem examinations revealed emaciated carcasses and considerable nematode burdens, with protein-energy malnutrition as the probable cause of death. Haematological and blood biochemical changes in the sheep, as well as fecundity of gastrointestinal nematodes, suggested the hosts were immunosuppressed. In parallel, laboratory experiments in which Soay sheep on a high plane of nutrition were artificially infected with Ostertagia circumcincta, showed no clinical signs or mortality when supporting worm burdens similar to those recorded in dead sheep on St Kilda. Anthelmintic treatment of a group of animals increased daily survival rates in ewes and male lambs, although treated animals became re-infected as the ‘crash’ progressed. It is suggested that parasites contribute to mortality in malnourished hosts, exacerbating the effects of food shortage.

Search behaviour of two entomopathogenic nematode species with different foraging strategies was compared by measuring parameters of unrewarded search after contact with host cues. Steinernema glaseri cruises in search of hosts. Steinernema carpocapsae ambushes hosts. Nematodes should respond to contact with relevant host cues by shifting their search from ranging to localized after contact with them. We predicted that cruising foragers rely on chemical cues more heavily than ambushers. These species were also tested for host affinities. Nematodes were tracked by image analysis after exposure to faeces, cuticle or food of either Popillia japonica or Spodoptera exiqua. Steinernema glaseri responded to selected host cues by shifting from ranging to localized search, characterized by decreased locomotory rate, distance travelled, search area and the proportion of the test period spent moving. Steinernema carpocapsae did not respond to host cues. Steinernema glaseri responds to selected chemical host cues for host location, whereas S. carpocapsae does not.

The influence of host dietary protein on acquired immunity and intestinal helminth population dynamics during repeated infection was studied using the mouse–Trichuris muris experimental model. CBA/Ca mice fed a 2% (by mass) protein diet ad libitum maintained body weight during the experiment, but when fed diets containing either 4% or 16% (by mass) protein gained weight steadily. Infection with T. muris did not affect the growth of the latter mice but significantly reduced the growth of animals fed on the 2% protein diet. When repeatedly infected with either 5 or 50 eggs every 10 days, the mice fed the 2% or 4% protein diet accumulated adults in proportion to infection dose. The results show that this is due to both the establishment of larvae at each repeated infection and the survival of established adults. In contrast, very few worms were recovered from animals fed the 16% protein diet, principally as a result of the development of strong acquired immunity to reinfection. T. muris egg output/mouse increased with infection dose in animals fed the low protein diets, but no parasite eggs were detected in the faeces of hosts fed the 16% protein diet. Mouse antibody reponses to adult worm excretory/secretory antigen were time- and infection dose-dependent in all 3 dietary groups. The major finding was that the specific antibody response was more intense, both quantitatively (serum OD levels) and qualitatively (antigen recognition by IgG1), in mice fed the low protein diets, even though they remained susceptible to infection. This study shows that host dietary protein deficiency, even at levels irrelevant to normal growth, can markedly potentiate the transmission of T. muris via alterations in host resistance. The high levels of antibody in susceptible animals suggest that this defect in resistance is unlikely to be due to nutrient deficiency-associated defects in humoral immunity.

Immunocytochemical techniques used in conjunction with confocal scanning laser microscopy (CSLM) and electron microscopy have been used to demonstrate, for the first time, the distribution of the parasitic platyhelminth neuropeptide, neuropeptide F (NPF) in the cestode, Moniezia expansa. Antisera were raised to intact NPF(1–39) and to the C-terminal decapeptide of NPF(30–39). These antisera were characterized and validated for use in both immunocytochemistry and radioimmunoassay (RIA). NPF immunoreactivity (IR) was detected using both antisera throughout all of the major components of the central and peripheral nervous systems of the worm. The pattern of NPF-IR was found to mirror the IR obtained using a C-terminally directed pancreatic polypeptide (PP) antiserum and FMRFamide antisera; blocking studies using these antisera revealed that FMRFamide and PP antisera cross-react with NPF(M. expansa). RIA of acid-alcohol extracts of the worm measured 114 ng/g using the C-terminal NPF antiserum and 56 ng/g using the whole-molecule-directed antiserum. While the C-terminally-directed NPF antiserum cross-reacts with NPF-related peptides from other invertebrates, the whole-molecule-directed NPF antiserum is specific for NPF(M. expansa). The C-terminal NPF antiserum has potential for use in the identification and purification of NPF analogues from other platyhelminth parasites.

Genomic DNA extracted from entomopathogenic nematodes isolated from 89 soil samples taken throughout the United Kingdom was hybridized with the ribosomal DNA clone from Caenorhabditis elegans (pCe7). When the DNA was digested with EcoR I and Hind III in a double digest, 5 distinct RFLP (restriction fragment length polymorphism) types were observed. While the prevalence of the 5 types was not equal, no correlation with geographical location, soil type or habitat could be detected. Subsequent hybridizations of total genomic DNA from the various RFLP types divided them into 2 groups. The most prevalent group, identified as Steinernema feltiae ( = bibionis), contained 2 of the RFLP types (Al and A2). The other group contained the remaining 3 RFLP types (B1, B2 and B3). Although similar to S. feltiae ( = bibionis), the members of the B-types can be distinguished from this species on morphological grounds and preliminary crossbreeding experiments have demonstrated that the 2 groups are reproductively isolated.

Potential mechanisms of immunoregulation have been investigated for the capacity to generate heterogeneity in the outcome of infection with helminth parasites. We have developed a mathematical model of the interaction between T cell and parasite populations, based on the assumption that activation of a Th1 CD4+ T cell response is required for host resistance. Antigen dose-dependent inhibition of Th1 cell proliferation generates heterogeneity in the outcome of host response to infection, with relatively low levels of exposure inducing resistance, and high levels of exposure associated with host susceptibility. Heterogeneity is additionally predicted in the duration of infection before individuals of the resistant class clear infection, with infection becoming more prolonged as the level of exposure rises. Similar categories of response are predicted if an alternative regulatory mechanism, that of interferon γ-regulated control of Th1 cell differentiation, is substituted into the model. However, the relationship between level of exposure and duration of infection is reversed. Results are discussed in the context of how these simple models of parasite–immune system interactions might be used to make predictions concerning specific examples of parasitic infection.

Proteinase activities were examined in extracts and excretory–secretory (ES) products of the infective and adult stages of the cattle lungworm, Dictyocaulus viviparus. Multiple enzyme activities were identified from each source, as defined by pH optima, substrate specificities, inhibitor effects and substrate gel electrophoresis. Serine-, cysteine- and metalloproteinases were identified, secreted materials being more active against protein substrates per unit protein than were extracts, and the particular proteinases produced varied with the developmental stage of the parasite. The antigenicity of these parasite proteinases was demonstrated by the inhibition of enzymic activity with Protein G-purified serum IgG antibody from both infected and vaccinated hosts and in the retardation of enzyme migration on electrophoresis of enzyme–antibody complexes. For the adult products, this confirmed that the enzymes concerned were of parasite origin, and not host-derived. These results argue for investigation of D. viviparus proteinases as targets for the antibody response in the limitation of parasite-mediated tissue damage and as the active principle behind the anti-D. viviparus vaccine.

Programmes to control Ascaris lumbricoides, Trichuris trichiura and hookworm infections have often been targeted at each infection separately, but the advent of benign and broad-spectrum anthelmintics suggests that combined control ma be feasible. The extent to which the infections co-occur in communities will determine the need for, and potential benefits of, such a combined approach. This paper examines the comparative distribution of the three geohelminths in different geographical areas and shows that A. lumbricoides and T. trichiura have closely related distributions, while hookworm infection is largely independent of the other two. These results indicate that many communities are at risk of disease from infection by more than one species of helminth. The similar distributions and epidemiological characteristics of A. lumbricoides and T. trichiura suggest that simultaneous control of these two parasites by the same strategy would be feasible and highly beneficial to communities. Multiple species control strategies which aim to control hookworm infection may require a more complicated protocol with more precise locality targeting.

This study describes the effects of testosterone (Te) on the intestinal nematode Heterakis spumosa in mice. The course of Heterakis infections is apparently under Te-control. At high circulating Te-levels as occurring in intact males, Te-treated females, and Te-treated castrated males, the period of release of Heterakis eggs in mouse faeces is greatly extended and the number of eggs released per unit time is markedly elevated in comparison to low Te-levels, as found in untreated females and castrated male mice. Also, the onset of the patent period occurs earlier in Te-treated mice. Testosterone also accelerates development and growth of both female and male worms of Heterakis in mice. Thus, young adult male worms can be observed in the upper colon of Te-treated castrated male mice on day 21 post-infection (p. i.), whereas, at that time, only L4 larvae are present in Te-untreated male castrates. Testosterone also favours the survival of nematodes in hosts. In untreated male castrates, the number of worms present on day 7 p.i. (L2 larvae) is approximately two thirds higher than that found on day 21 p.i. However, such a reduction in the number of worms does not occur in Te-treated castrated mice during the same period of time. The early phases of the life-cycle of Heterakis, i.e. hatching in the small intestine and final settling of L2 larvae in the upper colon are independent of Te. Also, Te does not affect motility and even slightly reduces the fecundity of adult female worms in vitro. Our data suggest that Te and/or Te-metabolites and/or Te-induced host factor(s) accelerate the development and growth of H. spumosa and favour the survival of Heterakis in the colon of mice.