The male intromittent apparatus of Ergenstrema labrosi and Ligophorus angustus, monogenean parasites of the thick-lipped grey mullet Chelon labrosus, consists of a slender flexible penis tube with an expanded proximal region and whose distal end passes freely through a canal in an accessory sclerite; the tube is surrounded by a sleeve of longitudinal muscle and the apparatus works in a manner generally similar to an engineer's Bowden cable. There is a substantial difference in the form of the accessory sclerite in the two species. In both parasites there is a slender sclerotized vagina which, in L. angustus, is short and straight or curved but in E. labrosi is about 7 times longer and forms a spiral. The special role of the sclerotization of the distal part of the vagina may be to resist perforation of the wall by the exceedingly fine penis tube and the consequent discharge of sperms into the parenchyma.

Rohdella siamensis, a new genus and species of aspidogastrean, is described from freshwater fishes in Thailand. A new subfamily, the Rohdellinae, is erected to accommodate this worm on the grounds that it differs from all known aspidogastreans in that the male and female ducts unite to form an hermaphroditic duct, with the result that the intromittent organ is a sinus-organ and not a cirrus. A new classification of the subclass Aspidogastrea is presented. Two orders are recognized: the Aspidogastrida, containing a single family, the Aspidogastridae, which in turn contains 3 subfamilies, the Aspidogastrinae, the Cotylaspidinae and the Rohdellinae subfam.nov.; and the Stychocotylida ord.nov., containing 3 families, the Stichocotylidae, the Multicalycidae fam.nov. and the Rugogastridae. The systematic position of the Stichocotylida, and especially the Rugogastridae, is commented upon.

A comparison was made between the early events (a sexual stages) in the life-cycle of Eimeria tenella in specifically immunized and control chickens. Particular attention was paid to the quantitative aspects and to the transport of sporozoites within intra-epithelial lymphocytes (IEL) from the enterocytes of the surface epithelium to the enterocytes of the crypts. There was a moderate decrease in the number of parasites initially seen in the mucosa of the immune birds, suggesting that some of the effects of immunity are exerted before penetration of the surface enterocytes, but the reduction in the numbers of developing parasites was more marked. This latter effect was due, at least partly, to failure of transfer of sporozoites from IELs to crypt enterocytes. These findings are discussed in relation to the efficacy of the immunity induced.

Fed adult Argas (Persicargas) arboreus produce a fecundity-reducing pheromone which affects all or most females in crowded conditions. Unfed adults and unfed and fed nymphs do not produce this pheromone. The effect of this pheromone is probably additive, increasing with additional crowding of adults. Mechanical disturbance is excluded as the cause of fecundity reduction. Fed females separated by a perforated barrier from crowded adults also exhibit very low fecundity. Adult excretory materials used 31 days post-feeding do not contain the fecundity-reducing pheromone. Preliminary experiments suggest that this pheromone is spatially limited in action, does not affect gonadotropic hormone synthesis and/or release, and does not induce oviposition deterrence behaviour. This may be considered to be a primer pheromone, causing alteration in normal vitellogenin synthesis and/or uptake and deposition in oocytes. The source and chemical nature of this pheromone should be investigated.

Isospora rastegaievae undergoes its asexual development in the intestinal tract of hedgehogs by endodyogeny. Using light- and electron-microscopy it was demonstrated that daughter organisms were formed by typical endodyogeny as described for different coccidia.

A 230000 molecular weight Plasmodium yoelii protective antigen was characterized. Two monoclonal antibodies against P. yoelii immunoprecipitated the 230000 mol. wt protein and a number of lower molecular weight polypeptides. These polypeptides were shown by peptide mapping and specific antibody binding to be fragments of the large protein. Iodination experiments suggested that the lower molecular weight species may be present on the surface of the merozoite. The protein was found not to be glycosylated. By serology, related antigens were shown to be associated with blood-stage schizonts of P. vinckei subspp., P. chabaudi subspp. and P. falciparum.

Feulgen-pararosaniline(SO2) staining was performed on stages in the life-cycle of Isospora (Toxoplasma) gondii, Eimeria tenella, Sarcocystis cruzi and Plasmodium berghei. The fluorescence emission of the stained DNA in nuclei of these stages was examined and compared with absorption microscopy measurements at 560 nm (green light) of the same specimens. Accurate identification of single cells, and especially discrimination between young schizonts and young gamonts was difficult after Feulgen staining. To overcome this problem preparations were first stained with Giemsa and the cells of interest precisely located with the aid of an England finder. The same preparations were then hydrolysed and stained with Feulgen—pararosaniline(SO2), after which the previously identified cells were investigated. The DNA distribution after Feulgen staining corresponded with the shape of nuclei after Giemsa staining. DNA was present in all investigated stages of the four parasites, including macrogamonts of I. (T.) gondii and E. tenella and peripheral blood gamonts of P. berghei. Macrogamonts could be recognized, even at a stage at which they can hardly be differentiated from young schizonts in Giemsa-stained preparations, by their typical distribution pattern of Feulgen fluorescence. Feulgen fluorescence was more granular and confined to the peripheral region of the nucleus, leaving a distinct nucleolus unstained. The horseshoe-shaped nuclei typical of macrogamonts could also be observed in some second generation merozoites of E. tenella, indicating that these merozoites are already sexually differentiated. The relationship between the present cytochemical observations and the ultrastructure of the four investigated parasites is discussed.

Mature asexual stages of the malaria parasite Plasmodium knowlesi synthesize proteins of Mr 180 000–225 000 that are expressed on the outer membrane of infected erythrocytes and which vary antigenically such that different parasite clones are specifically agglutinated with homologous antibody. Other non-agglutinable clones have been prepared which fail to express variant antigen on infected cells. Two agglutinable clones of different variant antigen phenotypes and a non-agglutinable clone were examined to determine the proportion of total malarial proteins represented by variant antigens. Malarial proteins were labelled with various radioactive amino acids and the sodium dodecyl sulphate—polyacrylamide gel patterns for the three clones compared by fluorography. The patterns were indistinguishable, the variant antigens being undetectable in analyses of total malarial proteins. Furthermore, these antigens were not detected by Coomassie Blue-staining of total cellular proteins after electrophoresis. Sodium dodecyl sulphate and Triton X-100 extracts of labelled cells were immunoprecipitated using a panel of sera of defined agglutination specificity. The variant antigens could not be detected in the fluorographic patterns of total malarial antigens immuno-precipitated by these sera. In contrast, after lactoperoxidase catalysed radio-iodination of intact schizont-infected cells, the 125 I-variant antigens on the cell surface were identified by demonstrating their accessibility both to antibody and to trypsin with intact cells. Thus, the variant antigens are quantitatively very minor malarial proteins that can only be detected by methods which selectively analyse the subset of proteins on the erythrocyte surface.

Borrelia crocidurae infection rates in different organs of adult Ornithodoros (Pavlovskyella) erraticus and O. (Ornithodoros) savignyi were studied at intervals during a 60-day period following an infective meal. The spirochaetes persisted in tick guts for 1 month, were first observed in other organs 1–7 days post-feeding, and persisted for 20–60 days in different organs. The ovaries, Gene's organ, testes and male accessory glands of only O. erraticus were infected. The infection rates in adult and immature O. erraticus and O. savignyi post-feeding on infected hamsters were 26·7–60·0% and 26·7–80·0%, respectively. Spirochaete trans-stadial transfer from originally infected larval O. erraticus to first-instar nymphs (N1) was 33·3%; to subsequent N2–N5 it was 40·0–60·0%. Infection rates in the resulting adults were 26·7% (males) and 46·7% (females). In O. savignyi, trans-stadial transfer from originally infected N1 to N2 was 26·7% and to subsequent N3–N7 13·3–33·3%; of the resulting males and females, 20·0% were infected. Transovarial transmission from female O. erraticus infected as adults occurred after the second gonotrophic cycle to 33·3–53·3% of the F1 larvae, and to 26·7 and 46·7% of the F1 males and females, respectively. All infected F1 females transmitted the spirochaetes to 26·7–73·3% of the F2 larval progeny after the first gonotrophic cycle. Transovarial transmission was not observed in O. savignyi.

Plasmodium knowlesi malaria-infected erythrocytes were radio-iodinated and several non-ionic, anionic and zwitterionic detergents were compared in their capacity to extract the labelled membrane proteins. The use of these detergents for antigen identification was tested by immunoprecipitation, after addition of Triton X-100 to some detergent extracts, using hyperimmune monkey antiserum and protein A-Sepharose. 125I-labelled antigens were specifically immunoprecipitated with all detergents tested, including the anionic detergents sodium dodecyl sulphate (SDS), deoxycholate and cholate; the zwitterions Zwittergent-312 and -314, CHAPS and Empigen BB, as well as several non-ionic detergents. The SDS-polyacrylamide gel electrophoresis patterns of 125I-labelled antigens varied after extraction with different detergents, there being no consistent pattern for detergents of a particular class. A total of 14 125I-labelled antigens were identified, 11 of them using Triton X-100. Some minor antigens identified with Triton X-100 were immunoprecipitated in greater amount after extraction in other detergents. Most importantly, two antigens Mr 200000 and 180000 were detected only after extraction with deoxycholate or SDS.

A technique is described for the growth of pure gametocyte cultures of Plasmodium falciparum. Using the classification of Hawking, Wilson & Gammage (1971) these cultures contain gametocytes of stages III, IV and V alone. Routine sexual cultures, varying from 0·35 ml static cultures to 500 ml shaking cultures, are exposed to an inhibitor of DNA synthesis (mitomycin C at 10 μg/ml) on the 11th day of culture, and the culture is harvested on the 14th day when capacitated stage V gametocytes are present. All other stages are killed by the drug and are morphologically degenerate within 2 days of the addition of the inhibitor.

Only a few years ago parasite immunology looked an unattractive subject better left to the dogged specialists. Parasites and hosts had been playing chess together for a million years, and there seemed little prospect of perturbing matters in favour of the host immune system. All that has changed, for three reasons. Firstly, we have learned how to grow at least some parasites in vitro, and prospects of doing so with others are encouraging. Secondly, progress in cellular immunology has revealed the sort of loopholes in the host defence system which parasites are likely to exploit: we are learning the questions which matter about parasites as antigens. Thirdly, and most importantly, molecular genetics is being brought to bear on parasites: we can now see a real, though long-term, prospect of manufacturing practicable vaccines through bio-engineering, and more immediately it gives us the tools needed to probe the host immune responses in the form of cloned antigens.

The behaviour of mice exposed to graded doses of Trichinella pseudospiralis larvae was monitored in an open-field situation. Ambulatory activity declined briefly during the acute phase of the infection (day 11) but regained pre-infection levels 1 week later. Animals receiving heavy doses of T. pseudospiralis exhibited a mild increase in the level of ambulatory activity during the chronic phase of the infection. The infections did not affect the level of exploratory activity. The effects of the T. pseudospiralis infections on the behaviour of the mouse host were much less pronounced than those induced by T. spiralis. It is suggested that these differences may be associated with the small size of T. pseudospiralis and the lack of capsule formation.

The endogenous development of the Houghton (H) strain of Eimeria mitis (= mivati) was compared with the life-cycle of a precocious (HP) line derived from the H strain. In both parasites 4 generations of schizonts which developed in epithelial cells were observed: the 1st and 2nd were found in the crypts and the 3rd and 4th in the villi. Gametocytes and zygotes occupied epithelial cells at the tips of the villi. The onset of gametogony normally coincided with the maturation of 4th-generation schizonts. The infection was confined initially to an area of the gut extending from the jejunum to the ileo-caecal junction but 3rd-generation merozoites and subsequent stages were also found in the caeca and rectum. The life-cycle of the precocious line was shorter than that of the parent strain. Gametocytes appeared to develop from 3rd-generation as well as from 4th-generation merozoites. Also, sporozoites of the precocious line transformed to trophozoites before those of the parent strain. First-generation schizonts of the HP line tended to be smaller and to contain fewer merozoites than those of the H strain. The differences between the life-cycles of the two parasites account for the lower reproductive potential of the precocious line.

The understanding of the mechanisms whereby intracellular parasites counteract the microbicidal processes of macrophages has progressed considerably in recent years. Various factors contribute to intracellular parasite destruction; from a biochemical standpoint, particularly important is the oxidative burst triggered by phagocytosis and by macrophage ‘activation’, that leads to the generation of toxic metabolites of oxygen. At the ultrastructural level, fusion of the parasitophorous vacuole with surrounding lysosomes appears to be a pre-requisite for the final digestion and elimination of the infecting microorganisms. The counter-measures evolved by microorganisms to escape intracellular destruction are best illustrated by studies in vitro on the interaction of parasites of the Leishmania, Toxoplasma and Trypanosoma spp. with mononuclear phagocytes. Some microbes are able to inhibit the fusion of phagosomes with lysosomes, thus avoiding the potentially harmful action of lysosomal hydrolases. Other microorganisms are able to resist the effects of such enzymes, perhaps by secreting inhibitory substances. Others still avoid lysosomes by leaving the phagocytic vacuole, to reach the cytoplasmic matrix where their development is unhindered. Particularly critical is the capacity of certain parasites to subvert the lethal effects of the oxidative burst. This can be achieved either by failing to evoke this metabolic response, or by producing scavengers that can detoxify harmful oxygen metabolites. Intracellular death or survival will thus depend on a delicate balance between the potency of macrophage cidal mechanisms, and the efficacy of the protective measures evolved by the infecting agents.

The migration in mice of 20, 50 and 90 krad. 60Co-irradiated Schistosoma mansoni larvae, biosynthetically radio-isotope labelled with [75Se]-selenomethionine, was evaluated by autoradiography of compressed tissues and compared to the migration of non-irradiated 75Se-labelled larvae. The migration of 20 krad. -irradiated schistosomula between skin and lungs was slightly delayed but otherwise paralleled the migration of normal, non-irradiated schistosomula during the first 8 days following exposure. By day 8 over 90% of both non-irradiated and 20 krad. -irradiated organises were located in the lungs. In contrast to non-irradiated organisms, however, only a small proportion of 20 krad. organisms migrated to the liver. The delay in migration between skin and lungs was more pronounced with 50 krad. -irradiated schistosomula. Nevertheless, 45–93% of 50 krad. -irradiated organisms migrated to the lungs by 8 days post-exposure. Over 90% of the 50 krad. larvae detected in the mouse on day 21 were in the lungs; no more than an occasional 50 krad.-irradiated organism was ever detected in the liver. In three experiments, over 85% of the 90 krad. -irradiated organisms were retained in the skin; in a fourth experiment about half of the 90 krad. -irradiated organisms migrated as far as the lungs. As with 50 krad. organisms, only an occasional 90 krad. organism was ever detected in the liver. Removal of the skin exposure site within the first 4 days of immunization with either 50 or 90 krad. -irradiated cercariae completely blocked the induction of resistance. Removal between the 4th and 6th days gave variable results. Mice had to be in contact with the irradiated larvae for a minimum of 8–11 days to stimulate a level of resistance comparable to that of mice whose immunization site was not removed.

The effects of the direction of gut flow, of injections of glucose and saline into different regions of the small intestine and of surgical re-routing or ligature of gastric, biliary and pancreatic secretions into the small intestine have been correlated with changes in the migratory response of the rat tapeworm Hymenolepis diminuta. Reversing the normal anterior to distal flow of luminal contents in the small intestine did not affect worm migration following feeding. Injections of a glucose-saline solution into the duodenum did not initiate a migratory response; similar injections into the mid- and posterior regions of the small intestine resulted in migrational responses similar to those following intragastric glucose feeding. Re-routing gastric secretions to the distal duodenum inhibited anterior migration of the worms beyond the new point of entry of gastric juices. Results following re-routing and ligation of the biliary and pancreatic secretions suggest that there is a potent cue to anteriad migration in the pancreatic secretions. Biliary secretions also appear to contain an additional migratory cue to worm migration. In order of importance the factors stimulating/inhibiting worm migration are pancreatic > gastric > biliary > glucose. The results support the hypothesis that the factors affecting worm distribution in the small intestine are interactive and synergistic, involve other luminal factors, such as 5- hydroxytryptamine and the physico-chemical gradients, and are of a regional nature such that the migratory response of a particular worm is directly related to its position in the small intestine when the cues to relocation are received.