The surface architecture of adult male Schistosoma mansoni was explored using a range of lectins with differing carbohydrate specificities. Highest specific binding was achieved with concanavalin A and the agglutinin of molecular weight 60000 from Ricinus communis; the binding of wheat germ agglutinin was mostly non-specific. Small amounts of peanut agglutinin and soybean agglutinin binding were observed and the binding of these lectins was increased by pre-treating the parasite with neuraminidase. The fucose binding protein of Lotus tetragonolobus failed to bind. These results indicate that mannose and/or glucose, galactose, N-acetyl-glucosamine, N-acetyl-galactosamine and sialic acid are exposed on the surface of the adult male schistosome.

Lactating and nulliparous outbred Swiss (CF-1 strain) mice were infected at 12–16 weeks of age with Nippostrongylus brasiliensis or Trichinella spiralis. Lysophospholipase B levels in the intestinal tissue and faecal pellets were greatly depressed in infected lactating mice in contrast to infected nulliparous mice. Correlated with these depressions in lysophospholipase levels were markedly reduced numbers of bonemarrow eosinophils in infected lactating mice. Although the peak levels of lysophospholipase in the intestinal tissues occurred in both nulliparous and lactating mice by days 9 and 14 after infection with N. brasiliensis and T. spiralis, respectively, lactating mice had significantly lower lysophospholipase peak levels than nulliparous mice. The peak of luminal levels of the enzyme coincided with peak levels of the enzyme in the intestinal tissues and with theexpulsion of the majority of the worms from the small intestines of nulliparous mice. However, lactation delayed the temporal relation between the peak of lysophospholipase levels in the intestinal lumen and worm expulsion. These results suggest that lactation depressed the levels of the enzyme and interfered with its release into the intestinal lumen.

The structure of the intestine of adult Nematodirus battus is described. It is similar tothe intestine of some other trichostrongyles but with a few minor differences. Changes in the structure of the intestinal cells are described in N. battus taken from lambs during the later stages of an infection (days 20 to 32) with 60000 infected larvae. The mostnoticeable changes were a decrease in the amounts of glycogen and smooth endoplasmic reticulum, a reduction in the numbers of secretory-type granules and the appearance of vacuoles within the cells. Some of these vacuoles contained small crystals, and crystals, some of them large, were also found in the intestinal lumen. These changes, with the exception of the appearance of the crystals, are similar to changes described for Nippostrongylus brasiliensis affected by the immune response of the rat host, and it is concluded that the changes in N. battus could have been initiated by an immune response to this nematode in the lamb. Lambs given 2000 larvae of N. battus do not reject the nematodes in 32 days, and no changes were observed in the structure of these nematodes.

An in vitro culture system incorporating a cell monolayer was used to induce most activated oncospheres of Taenia ovis to enter post-oncospheral development. Seven distinct developmental phases were recognized during the 23 days required for development to reach a stage similar to that achieved after 14 days in vivo. Secretions or tegumental substance were released during each developmental phase.

A simple in vitro technique was devised to culture chick embryos in Petri dishes from the 4th to the 21st day of incubation. Leucochloridiomorpha constantiae (Trematoda) metacercariae were placed either singly or multiply (5/embryo) on the chorio-allantois of in vitro grown embryos on day 7 and were removed on day 14. Growth and development studies were also made on worms grown singly or multiply (5/chick) in the bursa of Fabricius of the domestic chick. Worms grown singly or multiply in embryos were sexually mature, although eggs from these worms were abnormal when compared with eggs from worms recovered from chicks. The mean body area of worms from chicks was 2–3 times greater than that of worms from embryos. The mean body area of single worms from embryos was significantly larger than that of worms grown multiply in this site. However, the mean body area of multiple worms from the chick was significantly larger than that of single worms from this site. Worm pairs or clusters were seen in all embryos with the multiple infections.

The formation of the cyst wall surrounding Bucephalus haimeanus and the related changes in the parasite tegument during its metamorphosis from the cercarial to the metacercarial stage have been investigated by means of experimental infections in Gobiusculus flavescens. The initial fibrous cyst wall is formed from secretions produced by both the anterior gland cells and the tegument of the parasite. These secretions gradually become compacted against the surrounding hepatic cells until by the 20–30th day post-infection the 3-zoned inner layer, characteristic of the fully developed cyst wall, is formed. Hepatic cells immediately adjacent to this inner cyst wall layer are disrupted by the arrival of the metacercaria and form the middle vacuolated layer. As the metacercaria grows, the cyst increases in size causing still further hepatic cells to become flattened and incorporated into the cyst structure forming the outer nucleated layer. The structure of the cercarial tegument is described. Development of the metacercarial tegument is accomplished by the sequential movement of secretory vesicles from tegumental cell bodies into the outer cytoplasmic tegument. Vesicles of the types V. 1–5 are released from the outer cytoplasmic tegument, resulting in an almost complete breakdown of this layer prior to its replacement by the tegument of the fully developed metacercaria. The latter is characterized by the V. 6–9 type vesicles, dorso-ventrally flattened spines which terminate in 5–7 digits and by sensory cilia.

The daughter sporocyst of Cercaria vaullegeardi has hitherto been regarded as occurring within the haemocoel of the digestive gland of Gibbula umbilicalis. A re-examination has shown that an important part of the sporocyst, previously overlooked, extends from the digestive gland, through the afferent renal vein, the right kidney haemocoel and the transverse pallial vessel into the blood channel within a gill filament, thus providing a means of exit from the host directly into the mantle cavity. The sporocyst is described at light and electron microscope levels, and the emergence of the cystophorous cercariae is discussed.

The behaviour of oncomiracidia of the monogenean skin parasite Entobdella soleae, after hatching without chemical stimulation, has been studied in vertical, 4 cm high chambers illuminated from above. Most freshly hatched larvae are photo-positive and swim to the top of the vessel, where they remain actively swimming. Occasionally larvae become photo-negative and make a brief excursion to the bottom of the vessel. As the larvae grow older, these photo-negative excursions become more frequent and larvae spend progressively longer in the photo-negative phase of behaviour, but, even after many hours, photo-positive vertical excursions are still occurring. If this pattern of alternating photo-positive and photo-negative vertical movements occurs in the natural environment, these vertical movements, coupled with horizontal transport of larvae by water currents, would provide a search pattern for the host, the diurnally-inactive, bottom-living flatfish Solea solea. When eggs are stimulated to hatch in the chambers with urea, large numbers of larvae emerge within a few min, and it was observed that during the period of about 30 min after stimulation, many larvae are photo-negative. When eggs on the sea bottom are stimulated to hatch by mucus or urine from a nearby host, such behaviour would greatly enhance chances of contact with the fish. When eggs are stimulated to hatch by means of urea in the absence of light, there is evidence that the oncomiracidia are geo-negative. It has been shown that larvae are able to extricate themselves in total darkness from sediment containing buried eggs and that, in an experimental situation illuminated only by an infra-red beam, larvae readily attach themselves to the lower skin of soles within 2 min of hatching. There is evidence that, in aquaria, some invasion of the host takes place from below.

The reproduction and survival of the ectoparasitic digenean Transversotrema patialense on the fish host Brachydanio rerio are shown to be temperature-dependent. Survival is reduced by any deviation from an optimum temperature of approximately 23°C. With increasing temperature the rate of egg production per surviving fluke rises progressively faster to a higher peak up to 29°C but falls to zero by 35°C. The span of egg production varies closely with the life-span of the parasite at each temperature. The optimum temperature for the total number of eggs produced by each cohort of parasites is also 23°C. This number is a function of both adult parasite survival and the rate of egg output.

The egg-shell of Hammerschmidtiella diesingi and Aspiculuris tetraptera reduces the rate of water loss from the egg when exposed to desiccation. Staining of the enclosed larva by acid fuchsin and the collapse of eggs exposed to desiccation indicate that the permeability barrier is heat labile. This suggests that the lipid layer is the main permeability barrie of the shell. The complex structure of the uterine layers has a similar morphology to the respiratory structures of the chorion of some insect eggs. Reflections from the shell, the rate of embryonic development under various conditions and the penetration of lead ions indicate that the shell does not trap a layer of air when immersed in water and the uterine layers cannot, therefore, function as a plastron network. Calculations indicate that if diffusion is limited to the pores in the external uterine layer, the area available for gaseous exchange is reduced by 85·5% in H. diesingi, 95·6% in A. tetraptera and 96·8% in Syphacia obvelata. It is suggested that the uterine layers of oxyurid egg-shells function in a similar fashion to the pores in bird egg-shells and the aeropyles in the chorion of insect egg-shells which do not possess plastron networks. These structures reduce water loss from the egg whilst ensuring an adequate supply of oxygen for embryonic development.

Oocyst wall formation in Eimeria maxima was studied during the macrogamete stage in intestinal cells of the chick and in unsporulated oocysts isolated from faeces. The outer of the 2 membranes bounding the mature macrogamete separated from the surface but remained as a veil surrounding the developing oocyst throughout the whole intracellular process. Wall-forming bodies Type I were initially applied to the limiting membrane of the zygote cytoplasm; a layer of material similar to their contents was then formed around the zygote. As this occurred a new double membrane was formed surrounding the oocyst cytoplasm. The outer wall layer was initially homogenous in appearance but later developed into 2 zones, an outer amorphous region and an inner osmiophilic region. The inner layer of the oocyst wall was formed from the contents of the wallforming bodies Type II which dispersed between the outer wall and the limiting membranes of the oocyst cytoplasm. There was evidence of an additional membrane formed external to the outer wall. The outer membranes were not present around the wall of oocysts passed in the faeces of chicks, but the same wall zonation was evident, although the inner osmiophilic zone of the outer wall layer was markedly thinner in comparison with the same zone seen in the tissues.

The transmission of Hammondia heydorni among sheep, goats, cattle, dogs and coyotes is described. Oocysts of H. heydorni, obtained from dog faeces by feeding naturally infected goat meat, were fed to an experimental goat. After 312 days tissues from this animal were fed to 2 dogs. One dog, which received goat muscle, shed oocysts, whereas the other dog which received brain, spleen, kidneys, lungs and liver did not. Oocysts from the dog fed goat muscle were fed to an experimental sheep. After 73 days sheep tissues were fed to a dog and a coyote, both of which shed oocysts 5–8 days later. A second isolate of H. heydorni was obtained by feeding naturally infected beef to a coyote. These oocysts were fed to a single goat and sheep and after 71 days, muscle from these 2 animals was fed to 2 dogs and 2 coyotes, all of which shed oocysts 5–8 days later. None of the animals inoculated with H. heydorni became ill and no Hammondia-like organisms were found in tissue sections from the herbivores. Another dog shed H. heydorni-like occysts between 7 and 10 days after ingesting naturally infected muscles from a moose. These results indicate that H. heydorni infects goats, sheep, moose and coyotes in addition to cattle and dogs.

The sexual cycle of Theileria taurotragi was identified in the gut lumen of replete Rhipicephalus appendiculatus nymphs maintained at 28°C which had fed on eland with rising parasitaemias. Macro- and microgametes developed from ring-form piroplasms within 24 h after repletion. The microgamonts were elongate with 2 or more lateral projections. The nuclei of the microgamonts divided into 4 and the microgamonts differentiated into 4 thread-like microgametes each with a central nucleus. Round macrogametes developed at the same time and stages indicative of the fusion of the macro- and microgametes were observed after 48 h. The resultant zygotes were detected in the gut wall cells by day 4, many being rounded and vacuolated with a peripheral nucleus. By day 7 the zygote cytoplasm became dense and they lay in clusters still within the cells. Binucleate zygotes were observed at this stage. The zygotes increased in size and by day 12 began to transform into kinetes by invagination. By the time the nymphs moulted into adults (about day 14), the kinete straightened to a broad anterior end and a tapering posterior with a mean length of 22·1 μm. Kinetes were detected in the haemolymph by day 16. By day 20 the kinetes had penetrated the salivary gland acinar cells where they underwent schizogony until the infected acinar cells were filled with multinucleated sporoblasts. The nuclei of the infected acinar cells became greatly enlarged soon after penetration of the kinete. Division of the sporoblast nuclei was stimulated by feeding of the adult ticks. From the 2nd day of attachment of the ticks to rabbits the sporoblasts underwent a process of schizogony to produce cytomeres. Each nucleus of the cytomere divided to produce several small nuclei which differentiated into uninucleated sporozoites.

Strains of Babesia divergens of human and bovine origin were isolated from the same locality in Scotland. A morphometric comparison was then made between Babesia divergens parasites in blood smears made from groups of 3 splenectomized calves reacting to the inoculation of these strains. The various forms taken by the parasites of each strain were seen to be divisible into 7 categories and representatives of each category were found for each strain. More amoeboid forms were found in blood smears from calves infected with the strain of bovine origin but otherwise the frequencies of occurrence of different forms coincided. The dimensions of 3 forms of the parasites of each strain were compared and theparasites of human origin were found to be significantly larger. The reactions of the groups of calves to inoculation of the 2 strains of B. divergens were seen to be very similar, although inoculation of the bovine strain had a slightly greater effect on the animals' haematology as indicated by an increase in erythrocyte mean corpuscular volume.

A novel tetrahydroquinolinyl ester, quinfamide, administered orally in multiple doses for 3 days had an ED50, of 0·25 mg/kg/day (total dose 0·75 mg/kg) for eradicating Entamoeba criceti in hamsters in several tests. It was significantly more active by direct comparison than 3 commercially available amoebicides and at least as active as 2 other esters of the parent compound, 1-(dichloroacetyl)-1,2,3,4-tetrahydro-6-quinonnol. After administration of a single dose, ED50, calculations for quinfamide averaged 0·9 mg/kg. Quinfamide was considerably more active than the other tetrahydroquinolinols, diloxanide furoate and teclozan, and it was approximately 1·5 times more active than etofamide; a statistical significance between the latter 2 drugs could be demonstrated in one of 4 tests. Administered prophylactically, quinfamide was shown to protect hamsters from re-infection with E. criceti. It also inhibited propagation of E. histolytica in vitro at a concentration of 20 /tg/ml. No adverse effects were noted in rodents after a single dose as high as 10 g/kg. Daily administration to monkeys of doses up to 500 mg/kg for as long as 37 days produced no pharmacological aberrations during or after medication; haematological studies and urine analyses were normal and no gross or microscopical tissue changes attributable to quinfamide were observed. No toxicity was revealed following acute (2 g/kg) and chronic (500 mg/kg/day × 31 days) administration of the drug to dogs and rats, respectively.

A morphological study of the fine structure of the spores of Minchinia chitonis, a haplosporidian parasite of the chiton, Lepidochitona cinereus, is described. The spores contained a single nucleus, mitochondria, haplosporosomes, smooth endoplasmic reticulum, ribosomes and a large spherule (presumed Golgi apparatus). The spore wall was discontinuous at the spherule end, forming an opening covered by a lid which rested on a circumscribed flange. The flange of the spore wall and the lid were continuous in only one area which served as a hinge. The entire spore was encapsulated by epispore cytoplasm bounded by a strengthened membrane and extended to form 2 long projections, one at either end.

Studies were performed to identify specific parasite proteins synthesized within Plasmodium knowlesi-infected rhesus erythrocytes. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of whole parasites freed from the host erythrocyte by immune lysis, of membranous and cytoplasmic parasite fractions, and of isolated merozoites, detected several parasite-specific components after Coomassie Blue staining of the separated proteins. However, significant contamination with host erythrocyte material generally occurred, particularly in the whole parasite and parasite membrane preparations. Improved identification of plasmodial proteins was subsequently afforded by a radioisotope labelling technique in which parasitized erythrocytes were cultivated in vitro with [3H] isoleucine prior to electrophoretic analysis. Of 11 principal labelled peaks ranging in molecular weight from approximately 17000 to 145000 which were detected upon electrophoresis of whole parasites harvested from culture, all were observed in the cytoplasmic fraction while at least 5 were also associated with the membranous cell fraction. Analysis of different developmental stages of the intra-erythrocytic parasite revealed no significant stage-specific qualitative variations in the electrophoretic profiles. Quantitatively, however, the middle to late trophozoites incorporated more [3H] isoleucine into protein than the other intra-erythrocytic stages. Analysis of merozoites purified from labelled schizonts showed a protein pattern similar to the other stages. This confirmed that host components did not contribute to the labelling pattern and that none of the labelled proteins were specific to the residual cytoplasm remaining after merozoite formation.

Labelled antigens sharing determinants with both membrane and cytoplasmic fractions of Plasmodium knowlesi were detected in culture medium after in vitro incubation of schizont-stage parasites previously pulse-labelled with [3H]isoleucine. Release of antigens occurred only during schizont rupture and merozoite re-invasion. Antigenic material accounted for up to a third of the total trichloroacetic acid-insoluble radioactivity released by the cells. Absorption experiments indicated that approximately two-thirds of this antibody-precipitable material shared determinants with parasite membrane antigens, with a similar quantity sharing determinants with cytoplasmic proteins. Using antisera derived against 2 different antigenic variants of P. knowlesi, no evidence for the release of variant-specific antigens was obtained. Centrifugal analysis revealed that the majority of the radio-isotope labelled antigens were particulate in nature; however, some appeared to exist in true solution.

The expression of the virulence character in the virulent line (YM) of Plasmodium yoelii yoelii was investigated. The level of virulence was measured by counting the parasitaemia of the mature red blood cells. Several sub-clones were isolated from the virulent line YM and each was tested for its level of virulence. Out of 10 sub-clones 1 showed a marked decrease in virulence. However, transmission of this sub-clone through mosquitoes fully restored its virulence. A clone isolated from the progeny of a cross between mild and virulent parents had an intermediate level of virulence. A sub-clone isolated from this intermediate virulent line exhibited greatly reduced virulence. Mosquito transmission of this sub-clone also restored its virulence to a level comparable with the virulent line YM.

There is a need for more applied and experimental research to determine more precisely the relationships between nutrition, particularly childhood malnutrition, and intestinal parasitic infections, particularly Ascaris infection. Nevertheless, it is clear that in certain communities Ascaris infection is associated with poor growth in malnourished children and that deworming improves growth. Periodic deworming of children using a mass treatment approach may be needed to control soil-transmitted helminths in areas where parasites and protein energy malnutrition are highly prevalent. The main aims of treatment should be to reduce parasite loads below the level of clinical significance for the individual child and to reduce future environmental contamination with infective faeces for the sake of the community.