For the first time the development of the alimentary tract of Levinseniella brachysoma metacercaria (Trematoda: Microphallidae) obtained experimentally from Gammarus oceanicus has been described. The foregut primordium in 16-day-old metacercariae is represented by a syncytial cylindrical cord, resulting from the fusion of embryonic cells. Non-fused parts of the plasma membranes of adjacent cells are revealed as gap cavities within the cord. Later (30th day post infection) the lumen of the foregut is formed as a result of both partial vacuolization of the cytoplasm and by a broadening of the gap cavities, resulting from a thinning of the cytoplasmic spaces between them. Besides the usual organelles, the foregut of the mature metacercaria (42nd day p.i.) contains dense secretory granules in the apical cytoplasm region and numerous microtubules in basal areas. The cellular gastrodermis is formed later than the foregut syncytium (on day 30 p.i.); its large cells contain well-developed Golgi complexes, RER and mitochondria. A noteable inclusion of the gastrodermal cells of mature metacercariae are spherical granules of moderate electron density measuring up to 3 μm in diameter. On the basis of an analysis of the ultrastructural data the possible functioning of the metacercarial alimentary tract is discussed.

Experimental completion of the life-cycle of Paragonimus westermani (Trematoda: Troglotrematidae) (diploid type) from egg to adult has been achieved in the laboratory for the first time. A dog was fed metacercariae of P. westermani from naturally infected freshwater crabs, Geothelphusa dehaani. The adult flukes were recovered and their eggs were incubated in water at 28°C, hatching after 20 days. Laboratory reared marsh snails, Semisulcospira libertina, were exposed to miracidia from embryonated eggs. One of 47 snails became infected with a large number of 2nd generation rediae containing fully mature cercariae 245 days after exposure. Paragonimus-free crabs (G. dehaani) were then infected with the intramolluscan larvae. Of the 40 crabs examined 34–70 days after infection, 17 were infected with a total of 84 metacercariae. A cat and 4 dogs were fed with 55 metacercariae obtained. Eighty to 100 days later, a total of 40 flukes was recovered; 39 flukes were adults with eggs in their Uteri.

‘O’ group mullet, Chelon labrosus. were experimentally infected with Cryptocotyle lingua (Heterophyidae) by tail dip in a suspension of cercariae. Metacercariae were excised after 1 and 24 hours and prepared for TEM and post-embedding immunogold labelling. Antisera to cercariae of C. lingua were raised in adult mullet by natural infection via the skin and by intra-peritoneal injection of sonicate. The membrane-bound vesicles within the syncytial lining of the metacercarial excretory vesicle were found to be intensely antigenic with both antisera: the epidermal secretory bodies type 5 within the cystogenousglands gave a positive response. Penetration gland contents were not found to be antigenic with either antiserum. Discharge of the membrane-bound vesicles coinciding with both the reorganization of the lining of the metacercarial excretory vesicle and with cyst wall formation appears to be of significance in the initiation of the host immune response. That the term ‘excretory vesicle’ in Digenea may be a misnomer is discussed in the light of current information regarding the wide range of functions attributed to this structure.

Groups of mice (NMRI) were infected with 0.6 and 25 metacercariae of the intestinal trematode Echinostoma caproni. An enzyme linked immunosorbent assay (ELISA) was developed for measuring the specific IgM, IgG and IgA antibody responses in serum, small intestinal tissue and small intestinal lumen of the mice, by using a crude adult E. caproni antigen. In infected mice, significant levels of IgM were measured in the sera from day 14 after infection and of IgG and IgA from day 28 after infection. Early in the infection, the mean level of serum IgM was higher in 6-worm infections than in 25-worm infections. Late in the infection, higher mean levels of IgA were reached in the serum of mice with high than in those with low dose infections. The onset of appearance of antibodies in the tissue of the small intestine reflected the picture seen in the serum. For IgM, and to a lesser degree for IgG, the highest mean antibody levels appeared in the posterior sections of the small intestinal wall, where also the parasites were located. The mean level of IgA, however, was uniform throughout the length of the small intestinal tissue. High levels of specific IgA were detected in the lumen of the small intestine on day 28 after infection, especially in the anterior sections, where only few parasites were located. No specific IgM or IgG could be detected in the intestinal lumen on this day. The results are related to the intestinal location and the pattern of expulsion of E. caproni in the mouse host.

The life-cycle of Tremiorchis ranarum Mehra and Negi, 1926 (Trematoda: Plagiorchiidae) infecting Rana tigrina and R. cyanophlyctis is elucidated. Life-cycle stages, from egg to egg-producing adults, were successfully established in the laboratory. Life-cycle took more than two months for completion. Cercariae were found in the snail, Pila virens collected from paddy fields at Chelembra in the Malappuram district of Kerala during the monsoon months. Cercariae are of the xiphidiocercous type. Metacercariae occurred beneath the skin and in head, tail and body musculature of tadpoles of R. tigrina and R. cyanophlyctis. Adult frogs are refractory to cercarial infection. Frogs became infected when they fed on tadpoles containing metacercariae. Successive stages in the development to adult from the metacercariae were followed. Chronological synchronization in the reproductive activity of the host and the parasite is briefly discussed.

Three age groups of Bulinus natalensis, immature, mature but not yet egg-laying and mature egg-laying, were infected with miracidia of Schistosoma margrebowiei. The growth of infected, exposed but showing no signs of infection, and uninfected control groups, were examined at weekly intervals for ten weeks post exposure. Snails exposed to infection when immature, even in the group where no patent development ensued, showed a statistically significant reduction in growth and this was evident as early as 2–5 weeks post exposure. When infection occurred at the stage prior to egg-laying a significant reduction in growth was seen but only in the group which developed a patent infection. This was also the case for the egg-laying group although the reduction in growth was only significant in the final three weeks of the experiment. Thus in all three age groups, growth rate was reduced and the infected snails were significantly smaller at the end of ten weeks compared with the controls. The reasons for these effects are discussed.

A survey of larval helminths in freshwater snails of Kinmen was conducted from 1986 to 1987. Parasitological examinations of a total of 726 live snails collected from 25 loci revealed that 20 of 80 Bithynia fuchsiana were infected with metacercariae of Echinostoma gotoi and 36 with metacercariae of other echinostomes. Among 57 Radix auricularia swinhoei snails, 27 were infected with echinostomes and with metacercariae of other flukes. Of 20 Cipangopaludina chinensis, 18 were found with larvae of echinostomes. Larval trematodes were also found in three of 37 Austropeplea ollula and two of 87 Gyraulus spirillus. Third-stage larvae of Parastrongylus cantonensis were found in Ampullarius canaliculatus (5/103), Sinotaia quadrata (20/141), Hippeutis umbilicalis cantori (1/70) and Gyraulus spirillus (2/87). Segmentina hemisphaerula were not infected. Cercariae of Centrocestus formosanus, Haplorchis pumilio and xiphidiocercaria were found in three, two and two specimens, respectively, of 37 Thiara tuberculata.

Mouse (C3H) mucosal glycoconjugates were examined in normal small intestines and intestines infected with Echinostoma caproni, or E. trivolvis using six different fluorescein-conjugated lectins: Triticum, vulgaris agglutinin (WGA), Ulex europaeus agglutinin I (UEA-I), Ricinus communis agglutinin I (RCA-I). Glycine max soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), and Arachis hypogaeu peanut agglutinin (PNA). The expression of lectin-binding sites and the intensity of the binding of lectins in the mouse small intestines were changed by infection with the echinostomes. Specific differences in the reaction to glycoproteins were clearly observed between the mouse intestines infected with E. caproni and those infected with E. trivolvis. In E. caproni infection, binding of most of the lectins to the villi was remarkably reduced in accord with the villous atrophy and loss of goblet cells. In contrast, in E. trivolvis infection, the binding of WGA, RCA-I and DBA was reduced in the microvillar surfaces, but binding of UEA-I and SBA were unchanged compared to the control intestines. The lectin binding to goblet cells in E. trivolvis-infected mice mostly increased. These observations may reflect the marked increase in goblet cells and the less severe damage in the villi of E. trivolvis infection compared to E. caproni infection. Most of the glycoconjugates were slightly reduced in the hyperplastic crypts except for N-acetyl glucosamine. It is possible that glucose metabolism in the host intestines infected with E. trivolvis was activated. resulting in an increase in the rate of mucin synthesis as well as qualitative changes in mucus, thereby mediating the expulsion of the worms.

Adult Corrigia vitta (Trematoda: Dicrocoelidea) inhabit the pancreatic duct of the fieldmouse, Apodemus sylvaticus, where, in numbers, they may occlude the duct lumen and prevent the flow of pancreatic secretions. Enzyme histochemical and immunocytochemical techniques, in conjunction with confocal scanning laser microscopy, have been used to examine the localization and distribution of cholinergic. serotoninergic (5-HT, serotonin) and peptidergic components of the nervous system of the adult worm. All three classes of neuronal mediator showed a common pattern of staining, occurring throughout the central and peripheral nervous systems. Of the four peptide immunoreactivities (IR) demonstrated (pancreatic polypeptide (PP), peptide YY (PYY), substance P (SP), FMRFamide), PP-IR was the most predominant, occurring not only within the central ganglia and longitudinal nerve cords, but also in subtegumental plexuses and in fibres associated with the egg-forming apparatus. PYY and FMRFamide IRs were evident throughout the central and peripheral nervous systems; FMRFamide immunostaining, in particular, highlighted innervation of the ootype and immunoreactive cell bodies around the Mehlis' gland. Both SP- and 5-HT-IRs were restricted to the cerebral ganglia, ventral nerve cords and associated cell bodies. The distribution pattems of these peptides and 5-HT within the nervous system of C. vitta suggest they are likely to function as neuronal mediators. PP, PYY and FMRFamide may also serve in regulating egg production.

Acanthocheilonema viteae, Litomosoides carinii and Setaria cervi were found to actively synthesize proteins in vitro. Different centrifugation fractions and their TCA-precipitable fractions were assessed for the distribution of newly synthesized proteins. Penicillin and streptomycin inhibited the process in A. viteae. The synthesis in S. cervi was susceptible to puromycin, chloramphenicol, cycloheximide, neomycin and polymyxin B. The process in L. carinii was strongly blocked by puromycin while chloramphenicol had no significant effect.

The aim of this study was to examine whether there was an increase of suppressor T cells, relative to helper T cells, in the intestinal lamina propria of Hymenolepis diminuta-infected rats, a condition which might allow the parasite to survive for the life-span of the host. Lamina propria cells were isolated by enzymatic procedure. All lymphocytes were passed over nylon wool columns in order to remove B cells; then the T cells were purified by a panning technique using monoclonal anti-T cell antibodies. Changes in the mitogen-stimulated synthesis of 1gM, IgG and IgA by normal peripheral blood indicator lymphocytes, as measured by sandwich ELISA, was used as an index of help and/or suppression. No significant suppressor T cell activity was observed in the cultures, either from control or infected intestine.

In order to determine the prevalence of intestinal parasites among labourers from Thailand in Taiwan, the stool samples from 1364 Thai labourers were examined by the formalin-ether concentration method and 18·0% were found to be infected with one or more parasites. The infection rates of hookworm, Opisthorchis viverrini, Strongyloides stercoralis, Enterobius vermicularis, Ascaris lumbricoides, Hymenolepis nana, Capillaria philippinensis, Schistosoma japonicum, Echinostoma sp., Entamoeba coli, Giardia lamblia, and Entamoeba histolytica, were 7·6%, 7·0%, 3·2%, 0·5%, 0·3%, 0·1%, 0·1%, 0·l%,0·l%, 0·4%, 0·2%, and 0·1% respectively. The prevalence was highest among the 21–25 age group (24·8%). Among the 245 infected persons, 91% were infected with one parasite, 8% with two parasites and 1% with three parasites. The finding that parasitic infections are prevalent among Thai labourers demonstrates the need for control measures in foreign labourers in Taiwan and further studies to determine the susceptibility of O. viverrini to the snail hosts of Clonorchis sinensis.

Proteolytic enzymes present in extracts of third (L3) and fourth (L4) stage larvae and adults of the cattle nematode Ostertagia ostertagi were defined on the basis of pH optima and proteinase inhibitor sensitivity in spectrophotometric assays using azocasein and elastin-orcein as protein substrates. Evidence that different classes of proteinases are expressed in a stage specific manner was provided by the contrasting pH optima and inhibitor sensitivities shown by the enzymes in the different parasite stages. Stage specificity was confirmed by gelatin-substrate analysis. In addition, proteolytic activity was sought in the excretory/secretory products (ES) of the L4 following simple in vitro culture. Contrasting pH and inhibitor sensitivities as well as gelatin-substrate analysis showed that different proteinases were present in somatic L4 extracts and L4 ES products. The secreted proteinases may be useful targets for serodiagnosis or vaccination.

Conjunctivitis due to the trematode Philophthalmus in a 13-year-old Israeli girl is described. A single worm, probably a mature Philophthalmus palpebrarum, was detected on the palpebral conjunctiva of the upper eyelid of the right eye. Removal of the worm resulted in rapid abatement of the ocular symptoms. This is the first documentation of human philophthalmosis in Israel.

The pathomorphological effects of daily and weekly doses of liposomized albendazole on Trichinella spiralis larvae have been examined and compared during the course of muscle infections in mice. Treatment with three and six daily doses of the drug given by intraperitoneal injection to mice resulted in marked pathological alterations to encapsulated larvae, mainly in the walls of the capsules. There was a 5% efficacy against T. spiralis in mice given three daily doses of albendazole, and 38% efficacy in mice given six daily doses of the drug. The same doses of drug, when administered weekly, did not reduce the numbers of parasite larvae, and the application of drug at 6-weekly intervals resulted in a 5% reduction in numbers.

A comparative studs of the spectral, electrophoretic and isoelectric properties of the haemoglobins of three trematodes. Paramphistomum epiclitum. Gigantocotyle explanatum and Gastrothylax erumenifer was carried out. A high absorption in the beta band region indicates that trematode haemoglobins have high oxygen affinities. Electrophoretic mobilities of all trematode and their host haemoglobins were different. The isoelectric points of trematode haemoglobins were found to focus in the acidic range except that of G. crumenifer haemoglobin I. which focused at an alkaline pH.

Three nematode-trapping fungi, one Arthrobotrys oligospora and two Duddingtonia flagrans isolates, were fed to Ostertagia ostertagi-infected calves to test their ability to destroy larvae of this parasite in faeces and consequently to reduce the transmission of infective larvae to herbage. The fungi had previously been selected for their capability to pass the alimentary tract of cattle without losing growth and nematode-trapping potentials. Dung was collected from three calves each fed one of the three fungi and placed as 1-kg cow pats on a parasite-free grass plot together with control cow pats from a calf that was not given fungi. The cow pats contained comparable concentrations of parasite eggs. The two D. flagrans isolates were highly effective in that they reduced herbage larval infectivity by 74–85%. In contrast, A. oligospora did not show any effect in the present experiment. Field experiments will demonstrate if D. flagrans represents a potential organism for biological control of bovine gastrointestinal nematodes under practical agricultural management conditions.

Pyridoxine (vitamin B6) deficiency was induced in cotton rats which were then infected with the filarial parasite Litomosoides carinii. Embryogenesis was assessed microscopically in worms taken from pyridoxine deficient cotton rats and from various categories of control animals. Embryogenesis was retarded in worms from pyridoxine deficient hosts and more abnormal embryos were present in such worms than in those from control animals.

The Enzyme-linked immunosorbent assay (ELISA) was used to detect Ascaris suum antibodies in swine sera with adult body fluid (ABF) as an antigen. The assay was standardized with respect to the antigen concentration and serum and conjugate dilutions. Cross reaction was found between the antigen and the sera from the swine infected with Metastrongylus apri. The ELISA was more sensitive than the complement fixation test. Five protein peaks were obtained from ABF by gel filtration on Sephacryle S-300. Fraction I was the most specific. A 105 kDa protein in the fraction reacted with swine lgG in the serum of infected animals in Western blot analysis.

Distribution of some elements during development of endogenous daughter (hydatid) cysts of Echinococcus granulosus of buffalo origin was determined by atomic absorption spectrophotometry. These analyses showed that the distribution of copper and cobalt was highest in the smallest cysts. These elements gradually decreased in the cysts as they enlarged. The distribution of zinc, iron and manganese was very high in the smallest cysts in comparison to copper and cobalt, but these three elements decreased greatly in larger cysts. Iron and manganese were the only two elements found in very high concentrations in “thin-walled” cysts. Like all other elements both calcium and magnesium decreased as the cysts increased in size. In the thin-walled cysts magnesium increased by four times initially and this was followed by a two-fold “influx” of calcium in the “white-spots” in the above cysts. The significance of these findings is discussed in relation to earlier works.