An electron microscopical examination has been made of the fine structure and disposition of pancreatic polypeptide immunoreactive cells associated with the egg-forming apparatus in Diclidophora merlangi. The cell bodies are positioned in the parenchyma surrounding the ootype and taper to axon-like processes that extend to the ootype wall. The terminal regions of these processes branch and anastomose and, in places, the swollen endings or varicosities form synaptic appositions with the muscle fibres in the ootype wall. The cells are characterized by an extensive GER-Golgi system that is involved in the assembly and packaging of dense-cored vesicles. The vesicles accumulate in the axons and terminal varicosities, and their contents were found to be immunoreactive with antisera raised to the C-terminal hexapeptide amide of pancreatic polypeptide. It is concluded that the cells are neurosecretory in appearance and that, functionally, their secretions may serve to regulate ootype motility and thereby help co-ordinate egg production in the worm.

The activities of glucose-6-phosphatase (G6Pase), frucrose-1,6-bisphosphatase (FBPase), phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate carboxylase (PC) were determined in homogenates of adult Schistosoma mansoni worms and compared with the activities in homogenates of rat liver and rat skeletal muscle, tissues with a high and a low gluconeogenic capacity, respectively. All four gluconeogenic enzymes were present in S. mansoni. The enzymes were less active than in rat liver, but the activities of G6Pase, PEPCK and PC were at least an order of magnitude higher than in rat skeletal muscle whereas FBPase was approximately equally active in S. mansoni and in rat muscle. Experiments with 14C-labelled substrates or [14C]NaHCO3 failed to demonstrate the actual occurrence of gluconeogenesis in S. mansoni. Some possible other functions of the gluconeogenic enzymes were investigated. Experiments with inhibitors of PEPCK gave no indications that this enzyme was involved in the degradation of glucose. This was confirmed by 13C-NMR experiments which indicated that lactate was formed from phosphoenolpyruvate via the actions of pyruvate kinase and lactate dehydrogenase, and that PEPCK did not participate in the formation of lactate. Substrate cycling between fructose-6-phosphate and fructose-1,6-bisphosphate was demonstrated to occur in adult S. mansoni. This shows that FBPase participates in the glucose metabolism of this parasite.

Monoclonal antibodies raised against antigens present in the excretions/secretions (E/S) of larval Trichinella spiralis, polyclonal antibodies raised against E/S and antisera from rabbits and pigs infected with T. spiralis were used in conjunction with immunocytochemical techniques to detect antigens in sections of muscle from mice that had been infected with T. spiralis for 15 or 30 days. The antibodies recognized epitopes in the stichocytes, on the surface of the cuticle, in the lumen of the oesophagus and in the lumen of the intestine of encysted larvae. Monoclonal antibodies 7C2C5 and 1H7 and the polyclonal antibodies recognized epitopes in the cavity occupied by the larva, in the cytoplasm of the nurse cell, and in the hypertrophic nuclei of the nurse cell, but did not recognize material in the smaller nuclei of the nurse cell, in the cyst wall or in the surrounding muscle. Monoclonals 3B2E6 and 1D11G8B2, which recognized epitopes in the stichocytes and on the surface of the cuticle of the larvae, gave negative results with the cytoplasm and nuclei of the nurse cell. A polyclonal antibody raised against Trichuris suis recognized epitopes in the muscle and hypodermis of encysted T. spiralis but gave negative results with material in the nurse cell and nurse cell nuclei. The possibility that the antigen detected in the cytoplasm and nuclei of the nurse cell is produced by the stichocytes of the nematode and that it is controlling genes of the altered muscle fibre, either directly or indirectly, is discussed.

Many liver-invasive parasites cause extensive liver damage which may result in an impaired ability to catabolize endotoxin. The influence of endogenous endotoxin on the progress of liver-invasive parasitic diseases has been investigated in murine Mesocestoides corti infection. Invasion of liver tissue by tetrathyridia resulted in extensive parenchymal destruction with fibrosis. In association with this, undetoxified endotoxin, in potentially biologically active concentration, was found on peritoneal macrophages, 5 months post-M. corti infection. Host susceptibility was influenced by the Lps gene for responsiveness to lipopolysaccharide (LPS). The parasite burden of LPS-responsive (C3H/HeN) mice was significantly increased in the livers of these mice when compared to LPS-resistant (C3H/HeJ) mice. LPS reduced the ability of normal peritoneal macrophages to kill tetrathyridia, when co-cultured in vitro. LPS also abrogated the ability of recombinant interferon-γ (r.IFN-γ) to enhance macrophage larvicidal activity. These in vitro findings were confirmed in vivo. Daily intraperitoneal administration of LPS, at low concentration, caused a 4-fold increase in parasite burden in the liver, while r.IFN-γ at optimal concentration reduced parasite burden by 57%. Post-infection macrophages have previously been shown to be refractory to cytokine-activation for larval killing. In this report, we conclude that (1) this refractoriness may be due to the presence of undetoxified endotoxin on post-infection macrophages and (2) endotoxin may reduce host resistance by abrogating effector macrophage response to IFN-γ.

The relationship between faecal examination for egg output and worm burden of Opisthorchis viverrini in man of 181 autopsy cases from Northeast Thailand is described. Diagnosis of the parasite infection by stool examination for the presence of eggs was less sensitive than the worm recovery technique. Using Stoll's dilution and formalin–ether technique, no eggs were detected in the faeces of 20 cases harbouring low worm burdens (< 20 worms). The quantitative faecal egg count by Stoll's dilution technique showed a strikingly close positive correlation with the number of worms recovered (r = 0·96, P < 0·001) indicating a strong linear association between eggs per gram of faeces (epg) and worm burden. The number of epg per worm was inversely correlated to the worm burden (P < 0·001), suggesting that density-dependent constraints on fecundity could operate to restrict egg output in heavy infections. The accuracy of egg counts for estimating worm burden and its relevance to parasite epidemiological research are discussed.

Antigens were extracted from the epicuticle/cuticle of intact female Onchocerca volvulus using 2% 2-β-mercaptoethanol and 1% SDS. In Western blot analysis a human infection serum selected for its high antibody titre against whole worm homogenates did not recognize any component solubilized by 1% SDS. However, the same serum did bind at least 7 antigens among the material extracted with 2-β-mercaptoethanol. These antigens have apparent molecular weights (Mr) of: 15 000, 18 000, 28 000, 78 000, 98 000, 120 000 and 200 000. In ELISA using this preparation as target antigen, 151 out of 153 human infection sera gave positive results. An Onchocerca-specific IgG1 monoclonal antibody, designated Cam1, recognized the 28 000 Mr antigen, which is the most prominent antigen detected by Western blot analysis using human infection sera. In ELISA, using material affinity-purified with Cam1 as target antigen, 149 out of 153 human infection sera gave a positive IgG response. From a cDNA library three expressing clones were isolated with a rabbit serum raised against 2-β-mercaptoethanol solubilized material. One of these clones was recognized by the monoclonal antibody Cam1.

Three techniques for estimating the intensity of Opisthorchis viverrini infection in individuals from a Northeast That community are compared. Egg counts were determined using a quantitative formalin/ethyl acetate technique, worm burdens were estimated by expulsion chemotherapy and antibody levels were measured by ELISA. Log-transformed worm and egg counts were closely correlated (r = 0·80), suggesting that both measurements provide good assessments of relative intensity of infection. However, no Opisthorchis worms were recovered from 34 people with high egg counts; probably due to problems with the expulsion technique in some individuals. Examination of egg production per fluke indicated that each fluke contributed an average of 180 eggs per gram (epg) of faeces and fecundity was negatively associated with total worm burden. Serum IgG levels correlated significantly with Opisthorchis egg count (r = 0·61) at two independent assessments. Although significant associations were observed between antibody levels and echinostome infection, analysis suggested that these reflected independent associations between these two variables and Opisthorchis infection and age. We conclude that all three measurements are useful for epidemiological studies.

This study was designed to test the accessory function of macrophages after activation with products of Taenia multiceps coenuri. Activation was carried out by intraperitoneal injection of mice with coenurus fluid or protoscolex culture supernatant, and function was assessed by adding these macrophages in progressively increasing numbers to macrophage-depleted lymphocyte cultures transforming under the influence of plant mitogens or coenurus-fluid mitogen. In contrast to normal macrophages, which have a progressively enhancing action on the above reactions, parasite-activated macro-phages at similar concentrations were progressively inhibitory. However, low concentrations of the activated macrophages enhanced mitosis as well as, or better than, normal. Lymph node cells from injected mice showed abnormal response to macrophage-derived signals. In particular there was subnormal reaction to macrophages in the presence of coenurus mitogen. These results suggest that T. multiceps coenuri may survive in the host because of their ability to reduce effective interaction between lymphocytes and accessory cells.

The metabolism of 5-hydroxytryptamine (5-HT), octopamine (OA) and dopamine (DA) by adult male Ascaridia galli was investigated by incubating worms cut into 5 mm lengths with radio-isotope labelled amine. [14C]5-hydroxytryptamine and [14C]octopamine were taken up by the tissues and were metabolized to apolar products that co-chromatographed using both high-performance liquid chromatography and thin-layer chromatography with N-acetyl 5-hydroxytryptamine (NA5-HT) and N-acetyloctopamine (NAOA), respectively. N-acetylation was by far the most important reaction detected under these experimental conditions. A brief incubation of cut worms with [3H]dopamine resulted in the formation of a radio-isotope labelled metabolite that co-chromatographed with N-acetyldopamine (NADA) on reversed-phase high-performance liquid chromatography. The N-acetyltransferase activity towards 5-hydroxytryptamine and octopamine was detected in crude homogenates of male worms only when the co-substrate, acetyl CoA, was added to the reaction mixture. This enzyme activity appeared to be mainly localized in a 40 000 g supernatant fraction. The failure of previous studies to detect N-acetylation of biogenic amines in tissue homogenates of nematodes may have been due to the low levels of acetyl CoA present in these enzyme preparations.

Chub and eels were experimentally infected via intermediate hosts harbouring cystacanths, with Pomphorhynchus laevis alone, or Acanthocephalus anguillae alone, or simultaneously with mixtures of both species in varying proportions, and sampled at 7, 56 or 112 days post-infection. Examination of chub revealed that both species showed low establishment and growth rates, differing markedly from British field data, where chub is apparently one of the most important hosts, and preventing further meaningful experiments. Both species showed higher establishment rates in eels, but A. anguillae both grew faster and matured to a greater extent than P. laevis, reflecting field data. No evidence of any interactions, competitive or otherwise, was found in the mixed infections in eels at low or high infection levels. The results were interpretated with regard to British field data and current theories of the role of interspecific competition in structuring fish helminth communities.

In the absence of exogenous divalent cations, the isolated brush-border plasma membrane of Hymenolepis diminuta possesses alkaline phosphatase activity (APA). APA is stimulated in the presence of exogenous Mg2+ and inhibited by low concentrations of Zn2+ or high concentrations of Ca2+, and inhibition of APA by Zn2+ is reversed by both Mg2+ and Ca2+. APA is inhibited by ethylenediamine tetraacetic acid (EDTA), ethyleneglycol-bis-(β-aminoethyl ether) N, N′-tetraacetic acid, and 1, 10-phenanthroline in time- and concentration-dependent fashions, with EDTA being the most effective inhibitor. Following treatment with EDTA, APA is restored by Mg2+ and, to a lesser extent, by Ca2+, but not by Zn2+. Thus, APA represents a Mg2+-dependent enzyme that can be partly activated by Ca2+ but only in the absence of Mg2+.

Natural populations of 3rd-stage Nematodirus battus larvae were present on pastures in North Wales throughout the year; highest numbers were present in late spring, with smaller peaks occurring in autumn. Inter-site variation was observed in the timing and magnitude of these peaks. Hatching on experimental plots occurred 2 months to 2 years following deposition of eggs. Intraspecific and inter-site variation occurred in the timing, and inter-site variation occurred in the magnitude, of the mass hatch on upland and lowland experimental plots. Arrested 4th-stage N. battus were recovered from Welsh Mountain lambs. Percentage arrest and number of arrested worms was greatest during winter and early spring. The prevalence and intensity of N. battus infection in 1-, 2- and 3-year-old Welsh Mountain ewes was low. The plasticity exhibited in the parasite's life-history is discussed in relation to potential changes in the epidemiology of nematodiriasis.

Female sex pheromone reception was inhibited in a bioassay by treating males of the beet cyst nematode, Heterodera schachtii, with the sulphydryl reagent, mersalyl acid. This effect was blocked by the sulphydryl group protectant, dithiothreitol, indicating the occurrence of sulphydryl groups in the receptor molecules and a role for sulphydryl groups in pheromone reception.

Pine wilt is the most serious disease of native pines in Japan and potentially the most important nematode disease of conifers in the world. The pinewood nematode Bursaphelenchus xylophilus was found to be the causal agent. Difficulties arose with respect to the precise identity of some isolates of B. xylophilus and of similar species B. mucronatus and B. fraudulentus. Restriction enzyme analyses of repetitive DNA revealed bands specific for the species B. xylophilus, B. mucronatus and B. fraudulentus. Hybridization patterns obtained with unc-22 gene of C. elegans, clearly identified B. xylophilus, B. mucronatus and B. fraudulentus as well as the different geographic isolates of these species. Furthermore, it is possible to define the phylogenetic relationships between the different populations constituting the ‘pine wood nematode’ complex.

Using a C-terminally directed pancreatic polypeptide (PP) antiserum and immunocytochemical methods, PP-immuno-reactivity (IR) was localized throughout the central (CNS) and peripheral nervous systems (PNS) of the cestode, Moniezia expansa. In the CNS, immunostaining was evident in the paired cerebral ganglia (primitive brain), connecting commissure, and the paired longitudinal nerve cords that are cross-linked by numerous regular transverse connectives. The PNS was seen to consist of a fine anastomosing nerve-net of immunoreactive fibres, many of which were closely associated with reproductive structures. Radioimmunoassay of this peptide IR in acid-alcohol extracts of the worm measured 192·8 ng/g of PP–IR. HPLC analyses of the M. expansa PP–IR identified a single molecular form which was purified to homogeneity. Plasma desorption mass spectrometry (PDMS) of purified parasite peptide resolved a single peptide with a molecular mass of 4599±10 Da. Automated gas-phase Edman degradation identified a 39-amino acid peptide with a C-terminal phenyl-alaninamide. Examination of its primary structure shows that it displays significant sequence homology with the vertebrate neuropeptide Y superfamily, suggesting that this platyhelminth-derived peptide is the phylogenetic precursor. Neuropeptide F (M. expansa) is the first regulatory peptide to be fully sequenced from the phylum Platyhelminthes and may represent a member of an important new class of invertebrate neuropeptide.