A total of 100 specimens of the European shag Phalacrocorax aristotelis aristotelis were examined for anisakid infections in the south-east North Atlantic. The taxonomic status of the anisakid nematodes was studied by structural analysis using light and scanning electron microscopy. Two species, Contracaecum septentrionale and C. rudolphii A (Nematoda: Ascaridoidea), occurred in the European shag with high values of prevalence and mean intensities of infection. These constitute new host records for both parasites, widening their known geographical distribution. Although the histopathology associated with the infections indicated that the anisakids did not have the potential to cause bird death, the parasites are capable of contributing to and hastening avian mortality in the case of heavily-parasitized cormorants and when stressed by xenobiotics and/or through a synergetic effect by other bioagressors.

The status of Schistosoma sinensium (samples from Thailand and from Sichuan, China) relative to other species of the genus Schistosoma was investigated using DNA sequences from the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene (partial) and the nuclear ribosomal DNA second internal transcribed spacer 2 (ITS2). Trees inferred from these sequences place S. sinensium as sister to the S. japonicum group and suggest a basal position in the clade utilizing snails of the family Pomatiopsidae. The sequence differences between specimens of S. sinensium from China and Thailand are at least as great as between S. malayensis and S. mekongi. Schistosoma sinensium is probably best regarded as a species complex.

The validity and reliability of haematuria when used as screening criteria in community diagnosis of urinary schistosomiasis is presented. Between May and July, 1998, 1173 persons were screened for haematuria and examined for the presence of eggs of Schistosoma haematobium in their urine from all participating households in the Ozitem area of Bende Local Government Area, Abia State, Nigeria. Haematuria showed a sensitivity rate of 41.0% and specificity of 82.0% when used to identify cases of urinary schistosomiasis. Some factors that influenced the validity of haematuria as a diagnostic criterion are discussed. The use of haematuria amongst subjects in the first twenty years of their life is recommended.

Mucus samples from rainbow trout skin with or without infections by Gyrodactylus derjavini were tested for the presence of lectins reacting with mannose, galactose and lactose. The samples inhibited the binding of biotinylated lectins (from Canavalia ensiformis, Artocarpus integrifolia and Erythrina corallodendron, respectively) to microtitre plates with covalently bound carbohydrates (mannopyranoside, galactopyranoside and lactose, respectively). However, the inhibition of C. ensiformis and A. integrifolia lectins was slightly greater when mucus from infected (but recovering) fish was used, suggesting an increase of mannose and galactose binding lectins in fish skin exposed to parasites. As mannose, galactose and lactose are present on the glycocalyx of Gyrodactylus derjavini, it is suggested that lectins could play a dual role in interactions between fish hosts and their monogenean parasites. Thus, recognition between parasite and host and also host responses towards parasite infections could both, at least partly, involve carbohydrate-lectin binding.

The helminth fauna of the horned leaf frog, Proceratophrys appendiculata (Anura: Leptodactylidae), was studied from 17 adult individuals from an island in Rio de Janeiro state, southeastern Brazil. A total of five nematode species (Aplectana delirae, Cosmocerca brasiliense, Schulzia travassosi, Rhabdias androgyna and Physaloptera sp.) and an unidentified cestode were recovered from the digestive tract, lungs and gall bladder of P. appendiculata and all the nematode species are new host records. Aplectana delirae had the highest values for the prevalence and intensity of infection and can thus be considered a core species in the helminth community of P. appendiculata. Aplectana crossodactyli is synonymized with Aplectana delirae (). The helminth fauna of P. appendiculata presents characteristics typical of those in amphibians.

Collections of flounder, Platichthys flesus, at two sites on the tidal River Thames in 1994 and 1995 have, for the first time, revealed the sympatric occurrence of the freshwater and marine/estuarine strains of the acanthocephalan parasite Pomphorhynchus laevis. This natural co-occurrence of the strains has been employed to compare infection levels and a range of parasite attributes of the two strains under conditions of sympatry. At both Lots Road (Chelsea) and Tilbury the marine/estuarine strain was present at far higher infection levels than the freshwater form. In a detailed comparison of worms from Tilbury flounder, a range of differences was revealed between the two strains. In single strain infections in individual fish, freshwater and marine/estuarine worms had distinct but overlapping gut microhabitat use patterns, with the former having a central intestinal bias and the latter a bias for the posterior region of the gut. In mixed strain infections, niche contraction occurred so that no overlap occurred. Freshwater worms were larger and had more eggs, more ovarian balls, and a higher percentage of fully developed eggs than the marine/estuarine worms. These differences are thought to reflect intrinsic, presumably genetically determined, differences between the two strains as they occurred in the same fish host species collected at the same place and time. Apparent differences in strain reproductive potential in flounder in the tidal Thames are discussed in the context of previous studies and the intermediate host segment of the parasite life cycle.

Enzyme-linked immunosorbent assays (ELISAs) which detect specific antibodies to fluke cysteine proteinases have provided good sensitivity and specificity for the immunodiagnosis of trematode diseases. To detect specific antibodies without the need for purified proteinase antigens, an immunocapture assay using Protein A was applied for the immunodiagnosis of paragonimiasis and fascioliasis. ELISA plate wells were coated with Protein A, incubated with diluted patient sera, then incubated with a preparation containing fluke cysteine proteinases, excretory–secretory (ES) products of adult Paragonimus westermani or Fasciola sp. The activity of fluke cysteine proteinases bound on the wells was measured by adding fluorogenic peptidyl substrate, Z-Phe-Arg-MCA or Boc-Val-Leu-Lys-MCA. This assay detected specific immunoglobulin G to cysteine proteinases of P. westermani and Fasciola sp. by measuring proteinase activity on the plate wells. Patient sera showed significant high values of proteinase activity when the wells were treated with the respective homologous ES products, whereas the sera had low values after treatment with the heterologous ES products. The sera of patients with other parasitoses and uninfected healthy individuals also showed low values after treatment with the above fluke ES products. Thus, Protein A immunocapture assay, which detected IgG specific for fluke cysteine proteinases, provided a high sensitivity and specificity for immunodiagnosis of paragonimiasis and fascioliasis.

A protoscoleces/vesicles in vitro maintenance test with assessment of viability by eosin exclusion was used to evaluate the quantitative and qualitative activities of isoprinosine, its active component inosine and the dipeptide methylester L-Phe-Phe-OMe on isolated protoscoleces of Echinococcus multilocularis for 24 and 48 h. Isoprinosine and inosine showed dose- and time-dependent activity, the latter displaying a more rapid effect than the former. A high activity was shown with L-Phe-Phe-OMe, when compared to praziquantel. Ultrastructural alterations were much more striking with L-Phe-Phe-OMe, with an effect similar to that of praziquantel, whereas the chemotherapeutic activity of inosine and isoprinosine appeared to be directed against a metabolic target, with a lethal effect not immediately visible at the ultrastructural level. Thus, the previously reported in vivo activities of these drugs result largely from a direct effect on the parasite.

A dichloromethane extract of the seeds of Millettia thonningii (Leguminosae) which contains a mixture of isoflavonoids (predominately robustic acid, alpinumisoflavone and dimethylalpinumisoflavone) is known to have larvicidal activity towards the miracidia and cercariae of schistosomes and to possess significant molluscicidal activity. The present investigation has assayed the effects of this extract on the electron transport systems of isolated rat liver mitochondria. The extract was found to inhibit mitochondrial electron transport at Site I (NADH dehydrogenase) at concentrations of 30–159 mg l-1. Although the extract is not as potent an inhibitor at Site I as rotenone, a known inhibitor of NADH dehydrogenase, such observations could explain the molluscicidal and schistosomicidal activity of dichloromethane extracts of the seeds of M. thonningii.

Aminopeptidase-like activities in crude whole body extracts of the free-living nematode Caenorhabditis elegans and the plant parasitic soybean cyst nematode Heterodera glycines were examined. General characteristics including pH optima, heat lability, and inactivation of enzyme by organic solvent were the same for the two species. All developmental stages of H. glycines exhibited activity. In older females, activity was present primarily in the eggs. Affinity for the substrate L-alanine-4-nitroanilide was the same regardless of the stage examined, and was similar for the two species (Km=2.3±0.3 mM FOR C. ELEGANS AND 2.9±0.2 Mm for H. glycines). Nearly all (>95%) of C. elegans aminopeptidase-like activity was present in the soluble fraction of the extract, while H. glycines activity was distributed between the soluble and membrane fractions. Specific activities of the soluble enzymes were highest in C. elegans and H. glycines juveniles. The C. elegans enzyme was susceptible to a number of aminopeptidase inhibitors, particularly to amastatin and leuhistin, each of which inhibited aminopeptidase-like activity more than 90% at 90 μm. In H. glycines, aminopeptidase-like activity was inhibited 39% by amastatin at 900 μm. The apparent molecular weight of the soluble C. elegans enzyme is 70–80 kDa. Some activity in H. glycines is present in the 70–80 kDa range, but most activity (80–90%) is associated with a very high molecular weight (>240 kDa) component.

Recombinant protein was produced from the cDNA library of Trichinella pseudospiralis, which seemed to form part of the excretory–secretory (ES) products. The library was constructed from cDNA of muscle larvae at 1 month post-infection, and immunoscreened with antibody against T. pseudospiralis ES products. A clone, designated Tp21-3, contained a cDNA transcript of 657 bp in length with a single open reading frame, which encoded 172 amino acids (19617 Da in the estimated molecular mass). The predicted amino acid sequence of clone Tp21-3 had a similarity of 76% to that of clone ORF 17.20 (GenBank under accession number U88239) from T. spiralis. The recombinant fusion proteins encoded by clone Tp21-3 were produced in an Escherichia coli expression system and affinity purified. On Western blotting analysis, Tp21-3 recombinant proteins migrated at 40 kDa and reacted to antibody against T. pseudospiralis ES products and T. pseudospiralis-infected sera. Sera were developed against Tp 21-3 recombinant proteins, which reacted to a single band migrating at 21 kDa in crude worm extract and ES products from T. pseudospiralis on Western blotting analysis, and reacted with stichocytes of T. pseudospiralis on immunohistochemical staining.

In this study, the excretory–secretory products (ESP) of the daughter sporocysts of Vesicocoelium solenophagum (Trematoda) and plasma proteins of its host, Sinonovacula constricta were characterized by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and gelatin-substrate gel analyses, and the relationships between them were analysed using immunoblotting. Proteinase activity was detected in the ESP from daughter sporocysts of V. solenophagum. Some polypeptides of the ESP were found to be recognized by antiserum, raised against plasma from non-infected S. constricta, suggesting that the ESP may mimic host molecules (molecular mimicry). In contrast, neither the obvious proteinase activity nor the binding to the antisera was observed for the soluble proteins of daughter sporocyst, indicating that the ESP may play a important role in the parasite–host relationship. Although the plasma of infected S. constricta contained polypeptides that were similar to the plasma of non-infected bivalves, increased quantities of proteins at >170 kDa, 15 kDa and decreased quantities at 60 kDa were observed in the plasma of infected bivalves. Immunoblotting analysis revealed that the plasma of infected bivalves had a faint reaction with both anti-non-infected plasma antisera and anti-sporocyst antisera. These results indicated that the structure and quantity of some polypeptides from the plasma of infected bivalves had changed because of the infection with V. solenophagum. The polypeptides between the plasma of bivalves from a non-epidemic area and that from an epidemic area were similar, but the former had more polypeptides of 170–220 kDa and much greater proteinase activity than the latter, suggesting that the increased polypeptides of 170–220 kDa and the high proteinase activity in plasma may be favourable for protecting the host from being invaded by the parasites.

Extraintestinal migration patterns of Pharyngostomum cordatum (Digenea: Neodiplostomidae) were studied in experimental rodents such as mice, rats, and hamsters. When metacercariae isolated from grass snakes were infected orally to rodents, they penetrated the intestinal wall at days 2–3 post-infection (p.i.) and were discovered mainly in the diaphragm, intercostal muscles, and vital organ such as the lungs at days 7–28 p.i., without morphological changes. Interestingly, from several rodents which died suddenly at days 2–9 p.i., small to considerable numbers of metacercariae were found, not only in the lungs, but also in the heart and brain. Within the tissues, worms were freely motile until day 7 p.i., but later they were surrounded by host cells, and finally tissue cysts were formed. When metacercariae harvested from the snakes and intercostal muscles of rodents were infected orally to cats, they developed into adult flukes in the small intestine. The results show that P. cordatum undergoes considerable extraintestinal migration including the vital organs of its rodent hosts.

Cortisol production in fry of rainbow trout (367–456 mg body weight) infected with the ectoparasitic monogenean Gyrodactylus derjavini (mean intensities 4.7 and 4.9 parasites per fish) was studied at two temperature levels, 4–6°C and 11–12°C, respectively. Due to difficulties in obtaining an adequate amount of plasma from the small sized fish, the corticosteroid concentration was measured in the body fluid recovered (as supernatant) after sonication and centrifugation of whole fry. Infected fry at 11–12°C showed an elevated level of cortisol compared to uninfected fry. However, the cortisol concentration was lower than in fish exposed to handling stress. At 4°C, the cortisol level in infected fish compared to uninfected was insignificantly increased. The findings are discussed in relation to the role of monogeneans as inducers of secondary infections.

Analysis of mitotic and meiotic chromosome spreads of Aspidogaster conchicola (Trematoda: Aspidogastrea) reveals a diploid number of 10 for this species. Haploid sets containing 5 bivalents confirm the diploid number. The karyotype consists of one pair of comparatively long subtelocentric chromosomes and four pairs of shorter acrocentric elements. The results are discussed in comparison with existing data on chromosomes of aspidogastrean species and other related helminth groups.