In the present study ultrastructural immunocytochemical techniques have been used to localize the cells producing two soluble egg antigens present in the hatching fluid of Schistosoma mansoni, using two recently described monoclonal antibodies (MAbs). In the miracidium of S. mansoni, the one apical and the two lateral unicellular glands showed immunoreactivity for the egg antigens, using a post-embedding immunogold technique. Immunoreactivity was associated with the contents of vesicles for both the MAbs, although the intravesicular distribution was not identical. This difference in ultrastructural staining pattern provides additional evidence that the two MAbs are binding with different epitopes of the same antigen or with different antigens altogether. The finding of free unbound label around the embedded miracidium is consistent with secretion having been released from the glands.

Immunochemical techniques were used to determine the distribution, chemical characteristics and relative abundance of immunoreactivity (IR) to two native platyhelminth neuropeptides, neuropeptide F (NPF) (Moniezia expansa) and the FMRFamide-related peptide (FaRP), GNFFRFamide, in the trematodes, Fasciola hepatica and Schistosoma mansoni; the larger S. margrebowiei was used in the chemical analysis. Extensive immunostaining for the two peptides was demonstrated throughout the nervous systems of both F. hepatica and S. mansoni, with strong IR also in the innervation of muscular structures, including those associated with the egg-forming apparatus. The patterns of immunostaining were similar to those previously described for the vertebrate neuropeptide Y superfamily of peptides and for FMRFamide. Ultra-structurally, gold labelling of NPF- and GNFFRFamide-IRs was localized exclusively to the contents of secretory vesicles in the axons and somatic cytoplasm of neurones. Double-labelling experiments showed an apparent homogeneity of antigenic sites, in all probability due to the demonstrated cross-reactivity of the FaRP antiserum with NPF. Radio-immunoassay of acid-ethanol extracts of the worms detected 8·3 pmol/g and 4·7 pmol/g equivalents of NPF- and FMRFamide-IRs, respectively, for F. hepatica, and corresponding values of 4·9 pmol/g and 4·3 pmol/g equivalents for S. margrebowiei. Gel-permeation chromatography resolved IR to both peptides in discrete peaks and these eluted in similar positions to synthetic NPF (M. expansa) and GNFFRFamide, respectively.

Adult parasites of Schistosoma mansoni reside within vertebrate mesenteric veins where they consume immense quantities of host glucose after transporting the sugar through their surface syncytium or tegument. Previously we obtained cDNA clones encoding two functional facilitated diffusion glucose transporter proteins expressed by S. mansoni adult worms (Skelly et al. 1994). Antibodies specific for one transporter (SGTP1) have been generated against an extrafacial and an internal domain of the protein and used to localize the protein by light and electron microscopy. By light microscopy both antibodies stain a linear structure approximately 1–5 μm from the surface of the tegument of adult male and female schistosomes. Electron microscopic examination of frozen thin sections show binding of the antibodies to membranes in the base of the tegument and not to the membranes covering the outer surface or their invaginations. Analysis of the gold distribution suggests that the extrafacial domain is disposed toward the interstitial space beneath the tegument and the internal domain faces the syncytial plasm. The localization suggests that SGTP1 may function to transport free glucose from within the tegument and into the interstitial fluids that bathe the internal organs of these parasites.

In a population of copepods (Cyclops strenuus abyssorum), with a naturally high prevalence of infection with procercoids of Diphyllobothrium spp., no difference in body size was found between infected and uninfected hosts. However, a significant reduction in the reproductive capacity of infected females was observed, 87% of uninfected females having developed eggs in their gonads and sacs compared with 21% of infected females. The feeding rate of infected copepods was relatively high soon after infection occurred, but gradually decreased to less than half that of uninfected animals. Respiration rate was also lower in infected copepods. Infected copepods showed reduced motility and impaired escape responses, which is likely to make them more susceptible to predation.

Eight taxa have recently been proposed as being encompassed by the genus Trichinella on the basis of allozyme and biological data. In this paper we show that an analogous 8 taxon structure for this genus results from the random amplified polymorphic DNAs (RAPDs). Five 10-mer or 20-mer primers were used under different polymerase chain reaction (PCR) conditions to produce multiband RAPD fingerprints from muscle larvae of 40 isolates of Trichinella spp. The resulting RAPD data were analysed following the numerical taxonomic approach, and the resulting classification was compared to that derived from allozyme data. The agreement found between allozymes and RAPDs, while supporting the polyspecific structure of the genus Trichinella, confirms the potential of RAPDs as a tool for the detection of cryptic species. The selected primers were tested on individual muscle larvae in an attempt to standardize a RAPD assay for the routine identification of the 8 taxa of Trichinella. Only 1 of the 5 primers yielded reproducible fingerprints from the single larvae. Using this primer, the 5 species and the 3 other taxa of the genus Trichinella can be identified in a single assay without the need for massive in vivo parasite production.

A previous paper reported that the intake of Onchocerca volvulus microfilariae (mff) by different species of Simulium is essentially proportional to the parasite load in the skin of infected carriers. This paper examines the fate of the ingested mff in susceptible vectors to assess the relationship between parasite intake and infective larval output in blackfly species with and without well-developed cibarial armatures. Analysis is based on data from 3 onchocerciasis endemic areas: Guatemala (S. ochraceum s.l.), West Africa (S. damnosum s.l./S. sirbanum) and the Amazonian focus between South Venezuela and Northern Brazil (S. guianense and S. oyapockense s.l.). The data, which include published and unedited information collected in the field, record experimental studies of parasite uptake by wild flies maintained in captivity until the completion of the extrinsic incubation period. The relationship between L3 output (measured as the mean number of successful larvae/fly or, as the proportion of flies with infective larvae) and average microfilarial intake, was strongly non-linear. This non-linearity was best represented by a sigmoid function in case of armed simuliids (S. ochraceum s.l., S. oyapockense s.l.), or by a hyperbolic expression in that of unarmed flies (S. damnosum s.l., S. guianense). These results are compatible, respectively, with the patterns of ‘initial facilitation’ and ‘limitation’ described in culicid vectors of lymphatic filariases. A maximum mean number of 1–3 L3/fly was observed in all 4 vectors. It is concluded that O. volvulus larval development to the infective stage is regulated by density-dependent mechanisms acting at the early phase of microfilarial migration out of the blackfly's bloodmeal. Damage by the bucco-pharyngeal armature may also be density dependent. A hypothesis, based on this density dependence is forwarded to explain initial facilitation, so far only recorded in vectors with well-developed cibarial teeth. Our results provide quantitative support for the conjecture that chemotherapy alone is likely to have a greater impact on reducing onchocerciasis transmission in endemic areas where the main vector has a toothed fore-gut than in foci where the vectors have unarmed cibaria.

Snails, Elimia semicarinata, infected with Proterometra macrostoma were collected monthly in 1990 and 1991 from North Elkhorn Creek near Lexington, Kentucky, and kept on a 12:12 h light–dark cycle for 2 weeks. Cercariae emerging from snails were classified into 8 strains (I–VIII) based on differences in number and distribution of spined and spineless papillae on the tail. Cercariae also had unique patterns of emergence, swimming behaviour and infectivity in 4 species of sunfish. Of 513 infected snails collected in May, 339 had pure infections with the strain frequencies (% of 339): I, 46·6; II, 7·7; III, 12·1; IV, 8·8; V, 0·6; VI, 2·7; VII, 11·8; VIII, 9·7. In the multiple infections, 159 snails shed 2 strains, 14 shed 3, and 1 snail shed 4 strains simultaneously. A comparison of sunfish and parasite populations in Kentucky, Ohio and Michigan indicated that strain frequency in P. macrostoma is regulated by the species composition of the sunfish population.

The patch clamp technique was used to investigate the action of the anthelmintic drug, oxantel, on nicotinic acetylcholine receptor (nAChR) currents recorded from vesicles of the somatic muscle cells of the nematode parasite Ascaris suum. The amplitudes of the currents were analysed at different membrane potentials to determine the single channel conductance. Also the open and closed durations were measured to determine the kinetic properties of the activated channel. Oxantel activated single nAChR currents throughout a concentration range 10–100 μM, these currents were not observed with oxantel-free pipette solutions. The mean open time of the activated channels at a membrane potential of –75 mV and a concentration of 10 μM was 1·34 ms. At higher concentrations the open times were shorter and voltage sensitive, decreasing in duration on hyperpolarization, thus suggesting open channel block. The kinetics were analysed using a simple channel block model. The forward block rate, K + B, increased with increasing oxantel concentration but showed little increase as the membrane was hyperpolarized. K + B was 2·41×107 M−1s−1 – 50 mV and 2·64 × 107 M−1s−1 at – 100mV. The unblocking rate constant, K – B, did exhibit voltage sensitivity being 443·6 s−1 at – 50 mV and 86·8 s−1 at –100 mV. Thus the blocking dissociation constant KB (= K – B/K + B) was 18·5 μM at –50 mV and 3·3 μM at –100 mV. The simple channel block scheme was found to be insufficient to explain fully the observations made; reasons for this are discussed.

The action of two peptides isolated from the nematode Panagrellus redivivus, PF1 (SDPNFLRFamide) and PF2 (SADPNFLRFamide) have been studied on synaptic transmission in the motornervous system of the parasitic nematode Ascaris suum. Intracellular recordings were made from Ascaris somatic muscle cells and excitatory junction potentials (EJPs) elicited by stimulation of the ventral nerve cord. The EJPs were cholinergic as they were blocked by the Ascaris nicotinic receptor antagonist, benzoquinonium. PF1 caused a slow hyperpolarization, similar to the action of this peptide first reported by Bowman, Geary & Thompson (1990) and further characterized by Franks et al. (1994). The hyper-polarization was accompanied by a marked decrease in the amplitude of the EJPs with an EC50 of 311 ± 30 nM (n = 5). This inhibition is unlikely to be due to a post-synaptic site of action of the peptide as the muscle cell input conductance was not significantly altered by PF1 and furthermore the response to bath-applied acetylcholine was not inhibited by PF1 at concentrations up to 10μM (n = 6). PF2 also inhibited the EJPs in a similar manner to PF1. These studies indicate that both of the peptides isolated from the free-living nematode Panagrellus redivivus have biological activity in the parasitic nematode Ascaris suum. PF1 and PF2 have inhibitory actions in contrast to the predominantly excitatory actions of the Ascaris endogenous peptides AF1 (KNEFIRFamide) and AF2 (KHEYLRFamide). The potent actions of the Panagrellus neuropeptides PF1 and PF2 in Ascaris suggest that peptides with a similar or identical sequence may also occur in Ascarisand have an inhibitory role in the motornervous system.

The effects of mild to moderate infections of Trichuris trichiura on cognitive functions were investigated in Jamaican children aged 7 to 10 years. In all, 189 infected children and 100 uninfected classmates were studied. The infected children were randomly assigned to receive treatment (albendazole) or a placebo. All children were given cognitive tests on enrolment and 14 weeks later. These included verbal fluency (generation of ideas), digit span (working memory), number choice (speed of processing of visual stimuli), visual search (sustained attention) and a French Vocabulary test (paired-associate learning). At baseline, the infected children had lower scores than the uninfected ones in fluency (P = 0·01), search (P = 0·02) and French (P = 0·01). Treatment effects were examined among infected children and there was no significant treatment effect for any of the tests. However, there was a significant treatment by weight-for-age interaction in fluency (P < 0·05). The children with low weight-for-age (Z-score < – 1) improved with treatment while there was no improvement with treatment among the other children. We concluded that treatment of children with mild to moderate T. trichiura infections using albendazole produces little benefit in cognition if they are adequately nourished; however, undernourished children are more likely to benefit.

Proteinases released during in vitro maintenance of third (L3) and fourth larval stage (L4) and adult Teladorsagia circumcincta (formerly Ostertagia circumcincta), an ovine abomasal nematode parasite, were characterized on the basis of pH optima, molecular size and specific proteinase inhibitor sensitivity. Enzyme activity was maximal at alkaline pH and stage-specific release was demonstrated. Proteinases released by the adult parasite degraded a variety of protein substrates including plasminogen, albumin and haemoglobin, in a pH-dependent manner. At alkaline pH fibrinogen degradation was restricted to the α and β peptide chains although the γ peptide chain was also degraded at acidic pH. Inhibitor sensitivity studies indicated that degradation was predominantly due to metalloproteinases although aspartyl proteinase activity was indicated at acidic pH.

The timing of the energetic consequences of a developing, single-dose infection of Trichostrongylus tenuis larvae was investigated in captive red grouse Lagopus lagopus scoticus. At 12 days post-infection (p.i.), infected birds had a resting metabolic rate 16% greater than controls and thenceforth lost weight at a faster rate than controls. At 16 days p.i. infected birds consumed 38% less energy and excreted 33% less energy than controls. The estimated total daily energy expenditure and energy expended on activity for infected birds at 16 days p.i. were 36% and 83% lower, respectively, than for controls. Infected birds lost condition from 16 days p.i. onwards. The period of energy imbalance at 12–16 days p.i. coincided with development of late 4th-stage larvae into adult worms and the onset of patency. After this, the effects on energy balance diminished. Synchronous development of previously arrested T. tenuis larvae in wild birds in spring probably has similar effects to those reported here and places grouse under conditions of energy imbalance. The observed effects on energy balance provide a possible mechanism by which the parasite can reduce fecundity and survival of infected grouse.

The 4 species of ruminants (dromedary, zebu cattle, sheep and goat) in arid areas of Mauritania harboured Haemonchus spp. as the most frequent internal parasite. This was a rare situation where the 3 putative species, H. longistipes (dromedary), H. placet (zebu cattle) and H. contortus (sheep and goat) occurred sympatrically. The study was undertaken on hosts slaughtered at the Nouakchott abattoir, on the basis of monthly collection of worms. The environment was very unfavourable to H. placei and unfavourable to H. contortus, as intensity of infection remained low throughout the year, whereas infection in the dromedary was 10 to 20-fold higher. The survival strategies during the long, dry season were different: the surviving stages were either 4th-stage larvae in digesta (dromedaries), 4th-stage larvae either in digesta or mucosae (cattle), or 4th-stage larvae in mucosae and few adults (sheep and goats). The prolificacy of female worms, indicative of the potential to contaminate pastures, was similar for all Haemonchus spp. in the rainy season. H. longistipes behave differently during the pre-rainy season as no increase of prolificacy could be demonstrated as observed in the other species. Traits of vulvar morphology are considered as markers of ecological adaptation and were studied. The knobbed and smooth female morphs (in equal proportions) were the most frequent in H. longistipes, the knobbed morph out-numbered the other morphs in H. placei, and all 3 morphs were present in sheep and goats with the linguiform form being predominant. Genetic characterization of the 3 species was performed by means of Random Amplified Polymorphic DNA (RAPD). Three groups were obtained from analysis of these data: 1 group with individuals of H. contortus, 1 group with individuals of H. placei, and 1 group with individuals of H. longistipes. This indicated that, although the 3 species were valid, H. contortus and H. placei were more similar. Intraspecific variability was 2-fold higher in H. contortus than in the 2 other species. The ecological, morphological and genetical studies showed that H. longistipes, H. placei and H. contortus could be arranged in increasing order of variability.

In this paper the dynamics and control of nematode parasites of farmed ruminants are discussed via a qualitative analysis of a differential equation model. To achieve this a quantity, ‘the basic reproduction quotient’ (Q0), whose definition coincides with previous definitions of R0 for macroparasites, but extends to models with periodic time-varying transition rates between parasite stages or management interventions, is introduced. This quantity has the usual threshold property: if Q0 is less than one the parasite population cannot maintain itself in the host population, and in the long term becomes extinct; but if Q0 is greater than one the parasite can invade the host population. An alternative quantity, R(E), that is often easier to calculate is also introduced, and shown to have the same threshold property. The use of these two quantities in analysing models for the dynamics of nematodes in complex situations is then demonstrated, with reference to the dynamics of mixed parasite species in one host; the effects of breeding host animals for resistance to parasitism; and the development of parasite strains that are resistant to chemotherapy. Five examples are discussed using parameters for the dynamics of nematode infections in sheep, and some statements on control policies are derived.