Attenuated lines of Eimeria acervulina were isolated between 62 and 72 h post-infection from the Houghton (H) strain. The inoculation of small numbers of oocysts of precocious (HP) lines gave substantial protection to Light Sussex chicks kept on litter against challenge with the virulent H strain. The precocious trait of the 72 h HP line was shown to be stable because the kinetics of oocyst production remained unaltered after 9 consecutive passages through birds in which only late developing oocysts were used for passage. The precocious 62 h HP line was subjected to further selection for 5 consecutive passages. The resultant (62 A HP) line was shown to be more attenuated than the 72 h HP line but remained capable of immunizing chicks against challenge with the H strain.

By selection for early development of oocysts during serial passage through chickens the pre-patent period of the Houghton (H) strain of Eimeria mivati ( = mitis) was reduced by over 20 h. The precocious parasite is less pathogenic than the H strain and has a reduced reproductive potential. Chicks inoculated with the precocious parasite were protected against challenge with the H strain. The pathogenicity, immunogenicity and reproduction of precocious E. mivati and an attenuated egg-adapted line of E. mivati were compared. Although the precocious parasite produced more oocysts than the egg-adapted parasite in chickens, neither parasite induced loss of weight and both were immunogenic.

Eimeria mitis and E. acervulina affect different areas of the intestine. Young chicks received E. mitis with or without E. acervulina in doses ranging from 2 to 128 × 104 oocysts. Mixed infections produced significantly more severe effects on growth than single infections of either species. The ratio of villus height to total mucosal thickness was reduced throughout the intestine in birds which received the mixed infection. E. mitis and E. brunetti affect the same area of the intestine. Single infections of E. brunetti in a range of doses from 1.25 to 20 × 103 oocysts produced a similar reduction in growth. When 20 × 103 oocysts of E. mitis were inoculated simultaneously, the effect on growth was not so severe and the oocyst output of both species was reduced. E. brunetti, superimposed 1, 2 or 3 days after an E. mitis infection, produced no greater effect on growth than E. mitis alone. Similarly, infections of E. mitis, superimposed on E. brunetti had no greater effect on growth than the latter species alone. Oocyst output of E. brunetti in superimposed infections was not reduced but peak output was delayed.

The development of second generation schizonts of Eimeria necatrix and E. tenella was studied with the electron microscope. Invasion of the crypt epithelial cells by merozoites of the first generation schizonts caused changes in the morphology of the infected cells and stimulated their migration into the lamina propria through breaks which appeared in the basement membrane of the crypts. Second generation schizonts developed in the lamina propria within these crypt cells whose epithelial origin was confirmed by their interconnection by desmosomes and tight junctions and by their possession of characteristic microvilli. A comparison is made between this invasion of the lamina propria by parasitized cells and invasion of connective tissue by malignant epithelial cells; the possible mechanisms involved are discussed.

The microfibrillate component of the laminated layer of Echinococcus granulosus equinus contains, except for the zone adjacent to the germinal layer, aggregates of electron-dense bodies displaying a sub-structure of electron-lucent spheres. The tegumentary syncytial cytoplasm contains randomly distributed electron-dense granules, many occurring near the apical plasmalemma, although exocytosis was rarely seen. Granules, similar in size and sub-structure to the bodies of the aggregates, also occur in the internuncial connexions and tegumentary cytons, suggesting that they may be produced in the cytons and released into the laminated layer via the internuncial connexions and tegumentary cytoplasm. Cysts incubated for 0·5–2·5 h in serum- and non-serum-containing media showed differences from non-incubated cysts. The distal half of the syncytium contained a progressive increase in the number of granules (distal:proximal 1·9:1 at 2·5 h; cf. 1:1 in non-incubated cysts), and exocytosis of granules into the laminated layer adjacent to the cyst had occurred. This cannot be attributed wholly to serum proteins. Cysts incubated for 21 h appeared ‘normal’, suggesting re-establishment of an equilibrium. Since the matrix of the laminated layer is considered homologous to the glycocalyx of other cestodes, the possible protective role played by the granules/bodies, characteristic of Echinococcus spp., is discussed.

Peritoneal hydatids of Echinococcus granulosus equinus of 9 months standing in BALB/c mice occurred as free cysts or cysts within cyst masses. Both showed wide variation in size and in host tissue response, and all had a well-developed laminated layer separating the host tissue response from the germinal layer. In the smallest cyst-mass cysts the host tissue response was present as remnants of the initial cellular attack involving eosinophils. Slightly larger cyst-mass cysts possessed a primary macrophage invasion which phagocytosed the remnants of the initial attack and also, though to little effect, the laminated layer material. In the largest cyst-mass cysts a second macrophage invasion, of monocyte origin, had commenced and transformation stages of these cells to macrophages were observed. No fibroblasts surrounded individual cyst-mass cysts but they were present around the cyst mass, encapsulating it and possibly preventing further host cell invasion. Thus, the host tissue response around individual cyst-mass cysts remained ‘preserved’ at an early stage such as existed at the time of encapsulation. Small free cysts showed a primary macrophage invasion and transformation stages of cells of a secondary infiltration of peritoneal origin. Peripheral to the macrophages were fibroblasts demonstrating limited fibrinogenesis, and each cyst was surrounded by a layer of mesothelial cells. Large free cysts, also delimited by a mesothelial layer, possessed peripheral connective tissue, a deep fibrous layer and a monolayer of very compressed macrophages lying adjacent to the laminated layer. It is emphasized that an understanding of the host tissue response in cysts of different sizes and from different locations is an essential pre-requisite for the design of experimental studies.

Female Dermacentor andersoni were induced to feed on defibrinated bovine blood through fixed mouse skin membranes. Their feeding behaviour was recorded electronically, the tick being incorporated into the circuit and acting as a variable resistor during the periods of sucking, salivation and rest. Recordings of the feeding behaviour of these ticks were similar to those of ticks feeding on rabbits. Known concentrations of histamine, serotonin, dopamine, prostaglandin E1 and prostaglandin F2α were added singly or in combination to the feeding medium of ticks attached to the mouse skin membrane. Addition of both histamine and serotonin was followed by a significant reduction in the amplitude or recordings associated with sucking and salivation. The concentration of mediators producing this effect was less than 10 mM, a concentration comparable to that of histamine found locally in the skin of tick-resistant guinea-pigs. Comparisons are made between the effects produced by these mediators in ticks and the effects of serotonin and other amines known to produce modulation of complex behaviour patterns in other invertebrates.

Previous observations, which showed that treatment of mice with hydrocortisone acetate around the time of their infection with Schistosoma mansoni reduced the mature worm burden, have been confirmed. The number of eggs produced by the surviving worms during early patency was also significantly reduced. Cyclophosphamide and another steroid immunosuppressant, betamethasone, also caused a reduction in fecundity of S. mansoni when given at the time of infection, as did T-cell deprivation of the mice by adult thymectomy and injection of anti-thymocyte serum 1 month before infection. There was no effect of these three treatments on worm numbers. In contrast, injection of mice with anti-thymocyte serum at the time of infection marginally increased the size of the mature worm burden. The deleterious effects of hydrocortisone acetate on S. mansoni worm numbers and fecundity were only apparent if the steroid was given within 1 week of infection. Indomethacin, a compound which inhibits tissue inflammatory reactions by inhibiting prostaglandin synthesis, and which therefore mimics one of the actions of corticosteroids, had no effect on S. mansoni worm maturation and fecundity.

Changes during development in the elemental contents (Ca, P, K, Mg and S) of male and female Schistosoma mansoni from both mixed and single sex infections have been measured for the first time. In mixed infections, there is a significant difference between the calcium content of male and female worms; female worms having between 2 and 3 times more calcium. In mature female worms from mixed infections, after an initial rise in the calcium content corresponding to the time when vitelline development occurs, the amount of calcium ranged from 0.36 to 0.65 mg/g dry wt, whereas the amounts in similarly aged male worms ranged between 0.19 and 0.32 mg/g dry wt. While the type of infection does not affect the calcium content of male worms, a significant difference in the calcium levels in female worms from mixed and single sex infections was found. The amount of calcium in females from single sex infections ranged from 0.21 to 0.39 mg/g dry wt and this is considerably less than the levels recorded in females from mixed infections. This difference in calcium content between mixed and single sex infections can be related to the differences in degree of development of the vitelline gland in the two types of female worm. The amounts of phosphorus, potassium, magnesium and sulphur in both male and female worms from both single and mixed infections are similar, although females from mixed infections contained slightly more phosphorus than either females from single sex infections or either type of male worm.

After demonstrating that rats were capable of discriminating between the same diet treated with either flavour 1 or flavour 2 and that the 2 flavours of diet were equipreferred, an experiment was carried out to see whether learned taste aversion might play a role in the reduction of food intake that is commonly observed during the course of a parasitic infection. The results showed that rats, given a subcutaneous inoculation of approximately 6000 third-stage larvae of Nippostrongylus brasiliensis (Nematoda) while feeding on a diet containing flavour 2, strongly preferred to eat diet containing flavour 1 when given simultaneous choice conditions. Uninfected rats showed no preference and ate equal amounts of both flavoured diets. This effect is interpreted as the first experimental demonstration of learned taste aversion using a eukaryotic parasite as the inductive agent.

The synergistic relationship between host nutrition and the effects of Nippostrongylus was investigated experimentally in rats fed a diet containing 1% (w/w) protein. Rats were infected with 10 doses of N. brasiliensis varying from 0.35 to 99 larvae/g and their daily food intake and weekly body mass were compared with those of uninfected rats fed the same diet for a period of 9 weeks. The deterioration of the uninfected rats observed as a result of protein-energy malnutrition was found to be accelerated in a dose-dependent manner in the rats infected with N. brasiliensis. Analysis of food intake and body mass measurements demonstrated that this acceleration was related to a decrease in food intake and indicated a likely alteration in nutrient utilization, both of which were established by the end of the first week of the infection. Remission from the combined effects of malnutrition and infection was achieved by offering the rats a high-protein diet (20% w/w), whereupon rehabilitation was found to be complete within a period of 9 days. The implications of the results and the possible mechanisms involved in their generation are discussed in relation to the frequent coexistence of helminthiasis and malnutrition.

Plasma etching has been used to strip away the cortical layers of the cuticle of adult Nippostrongylus brasiliensis to reveal the struts, with their supporting fibres, which are found in the fluid-filled middle layer of the cuticle, and the basal fibre layers. The etched specimens were studied by means of scanning electron microscopy. The results support earlier work, obtained by transmission electron microscopy, on the cuticle of this nematode. Plasma etching has been shown to have potential in studying the structure of nematodes.

Males of Nematodirus battus have a body wall composed of a longitudinally ridged cuticle made up of 8 layers, a hypodermis and a layer of longitudinal muscle cells which are divided into 4 sectors by the hypodermal cords. There is a characteristic cephalic inflation of the cuticle in both sexes. The structure of the body wall in the anterior half of the female is similar to that of the male. In the posterior half of the female the cuticle is not ridged and has 7 layers; the longitudinal muscles are reduced in number and the lateral hypodermal cords increasingly extend dorsally and ventrally along the length of the posterior part of the nematode and come to occupy most of the lateral sectors of the body. The possible significance of these modifications in the structure of the female nematode are discussed.

Individual females of Gyrinicola batrachiensis produce 2 types of eggs: thin-shelled auto-infective eggs are produced in the ventral horn of the reproductive tract and thick-shelled eggs (transmission agents) are produced in the dorsal horn. Fine structure of oogenesis and egg-shell formation in the 2 horns of the reproductive tract were studied and compared. Early stages of oogenesis were similar in both horns but mature oocytes differed considerably. Those in the dorsal horn were larger than those in the ventral horn; they contained large numbers of lipid droplets, peripheral patches of glycogen and several types of cytoplasmic granules presumably acting as yolk or playing a role in shell formation. Mature oocytes in the ventral horn contained large amounts of glycogen, relatively few lipid droplets and large multivesicular bodies. Four shell layers formed around ova in the dorsal horn: a vitelline layer, a lipid layer, a chitinous layer and an outer protein coat similar to that described in other oxyurids. Only the vitelline layer formed around thin-shelled eggs. Thick-shelled eggs did not embryonate in utero but thin-shelled eggs nearest the vagina contained larvae. The first moult in eggs of G. batrachiensis was described in a previous communication and it is suggested here that the thin fibrous layer loosely applied to the cuticle of infective larvae in thin-shelled eggs is the moulted 2nd-stage cuticle.

Some unique features of monogenean biology have emerged from studies on 2 polystomatids which infect spadefoot toads in Arizona, USA: Pseudodiplorchis americanus in Scaphiopus couchii and Neodiplorchis scaphiopodis in S. multiplicatus and S. bombifrons. Transmission is confined to 1–3 nights each summer when the desert-adapted toads enter water to spawn. Encapsulated larvae complete development in utero and hatch within seconds of deposition; their body length (up to 600 μm) is 2–4 times greater than that of most other oncomiracidia, and their swimming life (over 48 h at 25–27 °C) is twice that usual amongst monogeneans. The ciliated oncomiracidia invade the nostrils, a target usually above the water surface and requiring migration over the exposed skin; remarkably they tolerate drying for up to 1 h (at 32 °C, 45% rel. hum.). The larvae migrate via the buccal cavity and glottis to the lungs where juvenile development occurs until early autumn. Migration from the lungs to the urinary bladder (by an as yet undetermined route) is followed by maturation and reproduction during host hibernation, and developing embryos accumulate in preparation for a period of transmission more restricted than that of any other monogenean. No other monogenean is known to exploit the lungs nor to inhabit, successively, anterior and posterior sites of the same adult host individual.

It was found that interference contrast microscopy gave good morphological differentiation of the living and intact salivary glands from adult ixodid ticks, Rhipicephalus appendiculatus and Amblyomrna variegatum, removed by dissection. In many cases the individual cell types of the acini could be recognized. When the salivary glands of infected adult ticks were examined Theileria parasites were detected. Theileria parva, T. taurotragi and T. mutans could be detected in the salivary glands after they reached the mature secondary sporoblast stage achieved even in unfed ticks. Parasites were detected only in type III acini. The morphology of tertiary sporoblasts and sporozoites of these Theileria spp. was clearly recognized in feeding ticks. Sporozoites of T. parva proved to be infective to cattle and to peripheral blood leucocytes of cattle in vitro after interference contrast examination. Interference contrast examination of living salivary glands was used for selection of salivary glands with high infections of T. parva sporozoites and the sporozoites could be separated in large numbers from these salivary glands. Using this technique it appeared possible to clone Theileria parasites from salivary glands with 1 infected acinus.

Sphaerospora molnari sp.nov. is described from the gills, skin and nasal pits of yearling carp (Cyprinus carpio L.) from ponds in the South Bohemian pond region of Czechoslovakia and one infected specimen from the river Danube in Bulgaria. Small vegetative stages, often massively pervading the infected tissue, produce 1 spore each. Spores are sub-spherical, 10·3 × 10·5 μm in size, with almost spherical polar capsules of equal size. The polar filaments are wound in wide coils that are almost parallel with the longitudinal axis of the spore. There are 2 uninucleate sporoplasm cells. Developmental stages and spores are also found in circulating blood. In the gills, S. molnari invades stratified epithelium of gill filaments and secondary lamellae causing branchial lesions and must be regarded as a serious pathogen of yearling carp. A new name, Sphaerospora chinensis sp.nov. is proposed for S. branchialis Lee & Nie in Chen (1973), pre-occupied by S. branchialis Razmashkin and Skrip-chenko, 1967.

Analysis of the seasonal occurrence of Argulus coregoni on Oncorhynchus masou and Salmo gairdneri indicates that the parasites overwinter as eggs and that there are 1 or 2 generations/year. Larvae which hatch from the overwintered eggs are abundant from May through July. Individuals hatched in May and June attain maturity and lay eggs in August. Larvae of the second generation are abundant in September, and they lay eggs in October–November which overwinter. Hatched larvae from overwintered eggs may mature in July and deposit overwintering eggs in September. The sex ratio of A. coregoni is about 1:1. A. coregoni shows an age-specific site preference on O. masou. Small parasites occur on almost all parts of the skin and fins, with slightly heavier concentrations on and around the pectoral and pelvic fins. Large parasites are found mainly on the skin around these fins.