The rediscovery of Elleipsisoma thomsoni Franca 1912 from Talpa europaea is reported. This organism, whose identity as a parasite was initially doubted, is shown to be an intra-erythrocytic protozoan parasite. Light microscope observation confirmed the original description and 22/53 (42·5%) moles examined from a locality near Blackpool were positive. Organ impression smears demonstrated that E. thomsoni was concentrated in the heart and lungs, occasionally in the kidneys, liver and spleen. Electron microscope studies conclusively showed that the parasite was a member of the phylum Apicomplexa. A typical apical complex was present with rhoptries, micronemes, polar ring and microtubules but, in contrast to the genera Plasmodium, Babesia and Theileria, a conoid was also present. The parasitized red cells were larger than normal mature erythrocytes and the remaining cytoplasm of the erythrocyte was less dense than that of non-infected red cells. The problem of studying this enigmatic parasite and its life-cycle are described.

Mice pre-treated with Corynebacterium parvum and later challenged with Plasmodium vinckei become infected but do not die whereas control mice do. When pre-treated mice were challenged with 1, 10, 1 × 102, 1 × 104, 1 × 105 or 1 × 106 parasites, the pre-patent periods correlated directly with the number of parasites injected, but the subsequent parasitaemias reached similar levels. This suggests that parasite killing, resulting from pre-treatment with C. parvum, is not triggered until the parasite load has reached a particular threshold. The injection of alloxan monohydrate, which brings about the release of toxic oxygen inter mediates thought to be involved in non-specific immunity, has little effect on P. vinckei infections until the parasitaemia is relatively high. This indicates that oxygen-mediated parasite killing also does not occur until the parasitaemia has reached a particular threshold. It is suggested that it is only at relatively high parasitaemias that the factors involved in parasite killing are able to enter the infected red blood cells.

The sheath and cuticle of microfilariae of Brugia pahangi were examined by electron microscopy and the presence of various proteins, carbohydrate and enzymes sought. The epicuticle of microfilariae consists of a pentalaminate structure (24·0 ± 1·4 nm), a cortex (13·7 ± 3·6 nm) and a basal zone (27·8 ±4·8 nm) which is often banded in appearance. The pentalaminate layers are not continuous at the base of the interannular grooves. The sheath and the epicuticle of B. pahangi stained positively with concanavalin A and saccharated iron oxide. The sheath of approximately 50% of microfilariae showed activity for acid phosphatase, 5′ nucleotidase and peroxidase, but not for ATPases, alkaline phosphatase or esterase. No enzymes were detected in the epicuticle although the cortex and basal layers of the cuticle did show enzymic activity. Structures beneath the cuticle in the main body of the worms contained considerable enzymic activity. Microfilariae directly isolated from the blood of infected cats were found by immunochemical means to carry serum proteins on their sheaths but not on their cuticles. These studies extend the definition of the outer structures of microfilariae and confirm that they significantly differ in morphology and enzyme content from typical mammalian cell membranes.

In vitro studies of the behaviour of the trypanosomatid flagellates Trypanosoma brucei and Leishmania hertigi in the presence of cell-free haemolymph of locusts, Schistocerca gregaria and cockroaches, Periplaneta americana revealed the presence of parasite agglutinins. The range of normal values of agglutination titres was 2−4 to 2−13. Physico-chemical treatment of haemolymph indicated that these agglutinins are protein or glycoprotein in nature and are only partially affected by heat treatment below 65°C, at which temperature incubation of haemolymph for 30 min abrogated all agglutination. Agglutination was not dependent on the presence of Ca2+ or Mg2+. Prior infection of locusts and cockroaches with T. brucei and L. hertigi significantly increased agglutinin titres between Days 4 and 6 in cockroaches (P < 0·05) and from Days 2 to 4 when L. hertigi was inoculated into locusts. The induced differences in titres observed in locusts infected with T. brucei were not significant. Lysozyme levels were significantly increased after inoculation of T. brucei into cockroaches compared with placebo-inoculated and uninoculated controls. L. hertigi inoculation produced significant increases in lysozyme levels compared with controls between Days 1 and 7 in locusts and 3 to 6 in cockroaches. These studies indicate that, at least in easily manipulated model systems, induced responses to intrahaemocoelic inoculation to trypanosomes and Leishmania can occur. As far as we are aware this is the first report of an induced response of an insect to such important parasites. The possibility that induced responses in natural vectors to these parasites occurs requires investigation.

SDS-PAGE analysis revealed that the protein composition of mechanically transformed schistosomula (ms) of a Japanese strain of Schistosoma japonicum was essentially similar to that of a Philippine strain of S. japonicum. However, the protein components of S. mansoni ms were remarkably different from those of S. japonicum. Specific proteins of Mr 60–65.and 30 kDa were found in ms of S. japonicum and S. mansoni, respectively. In skin-penetrated schistosomula (ss) of the two species, a common protein of 67 kDa was detected. Using two-dimensional electrophoresis, strain-specific polypeptides (9 in a Japanese and 11 in a Philippine strain) were identified in ms of S. japonicum. Mechanically transformed and skin-penetrated schistosomula of S. japonicum (Japanese) had, respectively, 7 and 10 specific proteins. Four and 1 specific polypeptides were also detected in two-dimensional profiles of ms and ss of S. mansoni, respectively.

The immune-dependent elimination of a challenge infection in rats vaccinated with highly-irradiated cercarriae of Schistosoma mansoni was analysed by passive transfer of serum, recovery of the challenge from the lungs and liver and by transferring lung-stage schistosomula. Recipients of serum from rats immunized with either unirradiated, 20 or 40 krad.irradiated cercariae, most of which die in the liver, lungs and skin, respectively, were equally resistant if the serum was injected on the day of infection or 5–7 days after infection. In addition, vaccinated rat serum transferred to mice and vaccinated rabbit serum transferred to rats conferred comparable protection when injected on day 0 or 5 days after infection of the recipents. This apparent susceptibility of the lung schistosomula to immune attack was confirmed by challenging 20 or 40 krad.-irradiated cercariae vaccinated rats with lung-stage schistosomula derived either from mice or rats. All the detectable attrition of a cercarial challenge in vaccinated rats occurred between 7 and 10 days post-challenge, before the parasites reached the liver. Since there was no evidence of damage or attrition in the skin or lungs before day 7 it was concluted that immune-dependent elimination occurred rapidly following a ‘window of sensitivity’ coinciding with the migration of the parasites from the lungs to the liver.

Intact surface membrane vesicles were obtained and purified from the schistosomula of Schistosoma mansoni. The method of preparation of the vesicles involved treating the schistosomula in a buffered high salt medium for 1 h at 4°C (Step 1), and then for 1 h at 25–37°C (Step 2). Vesicles were formed from mechanically- and skin-transformed schistosomula, and their size and number depended on the temperature of Step 2. The majority of the vesicles expressed surface membrane concanavalin A receptors and parasite antigens on their outermost surfaces. When incubated with sensitized peripheral blood mononuclear cell preparations from humans exposed to schistosomal preparations, the vesicles stimulated lymphocyte transformation as effectively as soluble egg and adult worm antigens. Although both preparations contained identical proteins, small vesicles were less effective than large vesicles in stimulating lymphocyte transformation. Hence, vesicles prepared under a variety of conditions might be used to study those factors which influence the presentation of membrane-bound surface antigens to the immune system of the host.

The outer tegument membranes of Schistosoma mansoni schistosomula have been removed by mechanical disruption in a hypotonic salt solution and partially purified by differential and sucrose density gradient centrifugation. Fractionation was monitored by measuring increase in specific activity of bound [125I] wheat-germ agglutinin (WGA), alkaline phosphatase and calcium-stimulated ATPase. Two-dimensional IEF/SDS polyacrylamide gel electrophoresis was used to analyse the peptide composition of the isolated membranes and to compare and contrast with lactoperoxidase/glucose oxidase surface labelled peptides. At least 35 surface-labelled peptides were resolved on the two-dimensional maps: all were also present in the membrane material recovered from the sucrose gradient. Western blot analysis demonstrated a marked heterogeneity in the antibody response of infected human patients to individual membrane antigens. The antigenic profile of membranes isolated from cercariae, 18 and 96 h schistosomula were compared using Western blots: some minor differences were observed between the three preparations.

The tegument of adult Schistosoma mekongi has been studied by both scanning and transmission electron microscopy. The gross surface topography of the parasite resembles that of S. japonicum, in that branched ridges, microvilli and sensory papillae predominate; such characteristics distinguish these two species from the non-oriental schistosomes. A unique feature of S. mekongi, however, is the numerous pleomorphic protruberances which are concentrated particularly on the middle three-fourths of the body surface. Transmission electron microscopy has revealed that these protruberances enclose bundles of microfilaments which appear to insert into the tegumental outer membrane. The microfilaments are suggested to have a supportive or stabilizing function, and may compensate for the absence of more typical crystalline spines. The tegumental outer membrane is typically heptalaminate in section, while the basal membrane infoldings are surrounded by concentrations of mitochondria. Three types of tegumental inclusion bodies have been recognized. Discoid bodies and membraneous bodies are morphologically identical to those described in all other schistosome species, except that the latter inclusions have been seen connected to each other and to the tegumental outer membrane by unique channels lined with trilaminate membrane. The third inclusion takes the form of spherical, lucent vesicles containing membrane fragments; these may represent the remains of spent membraneous bodies.

A stereological study was undertaken on the length, diameter and numbers of microtriches and on the structural and functional amplification factors of the adult cestodes, Hymenolepis diminuta, Hymenolepis nana, Eubothrium crassium, Proteocephalus pollanicoli, the plerocercoid of Schistocephalus solidus and the cysticercus of Taenia crassiceps. The study showed wide variations in these stereological parameters between species, stages and regionally in the same species. A comparison of the present results with previous studies has shown a large measure of agreement but some differences are evident and possible explanations of these differences are proposed.

A series of development stages (I–XI) have been devised to describe the development of the cyst wall of the metacestode of Hymenolepis diminuta. The cyst wall possesses tegumentary, muscular, fibrous and inner cyst tissues, the developmental rates and differentiation patterns of which are not identical. The tegumentary tissue differentiates posteriorly. Its microvillus-bearing distal cytoplasm remains simple until scolex retraction, after which rapid increase in depth followed by vacuolation occurs and basal membrane infoldings surround Phase 3 fibrogenesis fibrils. Senescence, which also affects the tegumentary cytons, then ensues. The muscle system development is posteriad and maturation, completed before scolex retraction, is followed by myocyton senescence. Posteriorly differentiated fibroblasts commence Phase 1 fibrogenesis after scolex retraction and the primary fibrous zone is fully established within approximately 6 days. Phase 2 and 3 fibrogenesis develop centrifugally, the fibrils of Phase 2 surrounding the tegumentary cytons and myocytons prior to their senescence, and those of Phase 3 lying more peripherally. The inner cyst tissue, established posteriorly, differentiates anteriorly, centripetally and early, the penultimate stage commencing just before scolex retraction, about 6 days after which the final maturation junctional complexes start development. Neither in vitro excystment nor infectivity of the definitive host can be satisfactorily achieved before the initial development of the primary fibrous zone. This may play a skeletal role during excystment, and is shown to be unaffected by the digestive enzymes which cause loss of cytoplasmic integrity in the outer regions of the cyst.

The physiology of the neuromuscular systems in Hymenolepis diminuta and H. microstoma was studied in vitro using intact, adult worm and strips of worm body wall. Intact worms were insensitive to ionic changes in the in vitro buffering system. However, strips of body wall containing longitudinal muscles were extremely sensitive to ionic manipulation. In intact worms tension generated in the strobila had two components; small brief tension peaks up to 500 mg amplitude are superimposed on larger, longer peaks of up to 1200 mg amplitude. Removal of the scolex and neck region either failed to show significant changes in tension, or showed a reduction in amplitude but not of frequency. Muscle contraction of both H. diminuta and H. microsoma were qualitatively similar. In split-worm preparations the concentration of Ca2+ in the bathing solution significantly affected both spontaneous and evoked contractions in H. diminuta and H. microstoma; the addition of CaCl2 greatly reduced the amplitude and frequency of the contractions. The chloride salts of cobalt, barium, cadmium and manganese elicited prolonged contractions of the longitudinal musculature of both H. diminuta and H. microstoma. While CoCl2 was the most effective in stimulating muscle contraction, the magnitude of the response varied with the concentration of Ca2+ in the bath. The results indicate that peripheral inhibition is extremely important in cestode motor control and that extracellular calcium ions may regulate the peripheral inhibitory mechanisms.

Male CD-1 mice inoculated by gavage with 400 larvae of Trichinella spiralis exhibited an increased latency of the wall-seeking response as well as a decline in running speed and the distance travelled to the first stop when compared with normal controls or animals inoculated with only 100 larvae. Doses of 200 larvae of this parasite affected running speed and distance travelled but not the advent of the running response. Such changes in behaviour may reduce the competitive fitness of the host and render it more susceptible to predation.

An experiment was conducted to examine, under laboratory conditions, the pathophysiology of chronic ovine haemonchosis. In the present study, groups of Merino and Scottish Blackface lambs were maintained on a low plane of nutrition and infected with 50 Haemonchus contortus larvae/kg live weight. The parasitized lambs along with respective breed controls were examined over a 27-week period. In addition to the body weight, haematological and parasitological parameters, erythrokinetic, metabolic, digestibility and nitrogen balance studies were conducted in the early stages of the infection. The results demonstrated that a low level of infection with H. contortus in lambs on a poor plane of nutrition caused the development of a normochromic normocytic anaemia which was associated with a modest but significant increase in abomasal blood loss and slightly elevated erythropoiesis in both breeds relative to the controls. The level of anaemia, hypoproteinaemia and pathophysiological changes were more marked in the Merino lambs and they also had higher faecal egg counts than the Blackface lambs.

During autumn and winter, separate pasture plots were seeded with Ostertagia ostertagi eggs produced by worms which had persisted in the host over summer as inhibited larvae, or which had developed directly from larvae acquired from pasture in late summer. Successive groups of parasite-free calves grazed the plots in spring for 14 days at 4-week intervals and were killed for worm counts 14 days after removal from pasture. The proportion of inhibited early 4th-stage larvae was substantially greater in calves which grazed the plot carrying the progeny of previously inhibited worms. This finding is more easily reconciled with differential survival on pasture over spring and summer of inhibiting and non-inhibiting morphs, than with the concept of a physiological switch from inhibiting to non-inhibiting behaviour in individual larvae.