A description is given of some features of the embryonic development, the structure of the oncomiracidium and the postlarval development of R. emarginata, a hexabothriid parasite on the gills of Raia clavata.

There are three phases in the larval development of the haptor of Rajonchocotyle: an oncomiracidial or marginal hook stage, a hamulus stage and a sucker stage. Neither the embryo nor the oncomiracidium ever possesses more than five pairs of marginal hooks and marginal hooks I (the posterior-most pair) are considered to be missing. The marginal hooks develop within distinct binucleate oncoblasts. During the early stages of post-oncomiracidial growth the secondary attachment organs of the haptor are formed and it is only after the completion of the development of the haptor that the reproductive organs begin to appear. Firstly, a pair of hamuli is acquired, followed by four pairs of suckers, which form in posterior-anterior succession at the site of marginal hooks III-VI. The first pair of suckers remains unarmed while the other three pairs acquire hooked sclerites and become the main functional attachment organs of the adult. As the first pair of suckers appears the posterior part of the haptor lengthens to form a caudal appendage. Hexabothriids are considered to show closer affinity to the 8-suckered chimaericolids and diclidophorids than to the 6-suckered polysto-matids, the unarmed suckers representing a simplification, by loss of the sclerite and reduction in sucker size, of an originally armed sucker.

I would like to express my thanks to Dr J. Llewellyn for his interest and for much helpful discussion.

Thanks are also due to the Director and Staff of the Marine Biological Association at Plymouth, in particular Mr J. E. Green for material and assistance, and to Dr A.j Brinkmann, of the Department of Zoology, University of Bergen, for supplying specimens of Squalonchocotyle borealis.

Carbohydrates associated with the tegumental surface of Schistosoma mansoni primary sporocyst may serve as potential receptors for mediating recognition by the internal defence system of the molluscan host, Biomphalaria glabrata. Therefore, a combination of SDS-PAGE and lectin probe analyses were carried out on biotin-labelled tegumental glycopeptides as a first step to defining the carbohydrates expressed at the sporocyst surface. The majority of surface polypeptides, ranging in relative molecular masses from 27 to 113 kDa, reacted with horseradish peroxidase-labelled Canavalia ensiformis (Con A), Erythrina corallodendron (ECA), Glycine max (SBA) and Triticum vulgaris (WGA) lectins indicating that most, if not all, tegumental proteins are glycosylated. However, differences in the binding of some lectins to individual glycopeptides suggest a degree of heterogeneity in the structure/composition of sugar moieties comprising these surface glycoconjugates. This notion is supported by the finding that the fucose-specific Tetragonolobus purpureas (TPA) lectin only reacted with approximately 50% of glycopeptides identified at the tegumental surface. Experiments employing biotin-labelled plasma (cell-free haemolymph) from S. mansoni-susceptible and -resistant B. glabrata snails as probes, further demonstrated that many of the identified surface glycoproteins also serve as plasma-binding sites for both snail strains. Binding interactions between plasma and sporocyst surface glycoproteins appeared to be, at least in part, mediated by carbohydrates since periodate treatment of sporocyst proteins or pre-incubation of plasma with the glycoproteins, fetuin or mucin, resulted in a decrease in plasma reactivity to blotted larval proteins.

Surface aminopeptidase (APase) in the acanthocephalan, Moniliformis dubius, was studied using leucylleucine, leucylglycylglycine, glycylglycine, alanylalanine, tri-alanine and tetra-alanine as substrates. In the presence of intact worms all peptides, except glycylglycine, were hydrolysed liberating amino acids and/or peptide subunits into the incubation medium. In 5 min incubations with 5mM leucylleucine, 93% of the liberated leucine was absorbed by the worms indicating a kinetic advantage for absorption of the leucine. In media with leucylleucine plus 10 mM methionine as an inhibitor of leucine uptake, the liberated leucine was not absorbed by the worms, but accumulated in the incubation medium.The inhibitory effects of leucylleucine and alanylalanine (or products of hydrolysis) on the uptake of 0·1 mM [14C]leucine in 2 min incubations were examined. Inhibition of [14C]leucine uptake by these peptides was non-linear up to 5 mM inhibitor concentration. Concentrations of either peptide greater than 5 mM did not produce further inhibition. A significant portion of [14C]leucine transport was not inhibited by these peptides. Inclusion of 0·5 mM Pb2+, an APase inhibitor, blocked the inhibition of [14C]leucine uptake by leucylleucine indicating that the inhibition is caused by the liberated leucine alone, and that interaction of the intact dipeptide and the leucine transport system is negligible.It is concluded that the surface membrane of adult worms has APase which can hydrolyse peptides in the ambient medium. The spatial arrangement of the APase and the leucine transport system is such that the hydrolysis of a peptide confers a kinetic advantage for absorption of the liberated amino acids.Encysted or mechanically excysted cystacanth larvae from the haemocoele of the cockroach showed no APase activity. Activation of the APase required a 30 min treatment in lipase, [Na+]taurocholate or other surface active agents. Since the APase in adult worms was unaffected by washing, the data strongly suggest that this enzyme is of parasitic origin.

Haliotrema balisticus attaches itself between the secondary gill lamellae of its host, Balistes caprisctis (= carolinensis), by means of a pair of ventral hamuli, which point ventrolaterally, and a pair of dorsal hamuli which point in a dorsal direction. The ventral hamuli, each of which pivots at the end of a ventral connecting bar, are operated by a pair of extrinsic muscles. The tendinous regions of these muscles pass through fibrous loops attached to the ventral hamuli and then join each other in the mid-line. Such an arrangement has a high mechanical advantage which may reduce the expenditure of energy required to maintain the hamuli in position in the gill tissue in the face of a strong gillventilating current and may also preserve the mobility of the free region of the body.

The dorsal hamuli are not operated by the muscles and tendons associated with the ventral hamuli.

Marginal hooklets and glands play subsidiary roles in attachment.

I wish to express my thanks to Commander J. Parreira, Director of the Aquario Vasco da Gama in Lisbon, and to the Staff of the Aquario, in particular to Mr A. Ferreira, for their kind assistance and hospitality. I am also grateful to Dr Maria Monteiro and to Dr Maria Baptista. I am indebted to the Calouste Gulbenkian Foundation for a grant which made my visit to Portugal possible.

An analysis of the rostellar hooks of Australian isolates of Echinococcus granulosus revealed that there was less variation in larval (metacestode) than adult characters and that metacestode characters could be measured directly from adult worms. A factor analysis indicated that two factors, one representing a contrast between number of hooks and their length, and the other representing blade lengths, were sufficient to account for 87.5% of the variance in metacestode hook measurements. These results indicate that rostellar hook morphology is not useful for discriminating strains of E. granulosus in Australia. The Tasmanian and mainland domestic strains were found to be indistinguishable using rostellar morphology. Although many of the isolates from sylvatic hosts differed from those from domestic hosts, there was not a clear separation as would be expected if two distinct strains existed. Evidence was presented to show that the morphological differences seen in sylvatic hosts could be attributed to host-induced effects, and that the previously accepted existence of two mainland strains should be investigated further.

Immunity to Hymenolepis nana was adoptively transferred to recipient mice of 4 strains using mesenteric lymph node cells (MLNC), spleen cells (SC), and/or serum taken from infected donor mice. Donor cells capable of transferring immunity appeared in the mesenteric lymph nodes and spleen at an early stage (5 days) after infection. Immunity was most effectively transferred with MLNC and immune serum did not enhance this effect. It is suggested that effective immunity to H. nana involves both antibody and T cell-mediated inflammatory events.

The effect of different relative humidities on the survival of individual larvae and adults of Ditylenchus dispasci and D. myceliophagus has been examined; experiments were carried out at 5 and 18°C with D. myceliophagusand at 18°C with D. dipsaci. Survival of both species was enhanced when they were dried at high humidities and, for D. myceliophagus, by the lower temperature. The survival of all stages of D. myceliophagus was poor. Generally it was expressed in minutes only; 4th-and 3rd-stage larvae, however, were marginally better at surviving drying than adults and much better than 2nd-stage larvae.

Although the 4th-stage larva of D. dipsaci was overwhelmingly superior to all other stages, the 3rd stage, and to a lesser extent, the adult of this species also showed remarkable powers of survival to desiccation. In general, survival of 4th-, 3rd- and 2nd-stage larvae could be expressed in weeks, days and minutes respectively; for adults, survival was expressed in hours at relative humidites under 50% but in days at higher relative humidites.

The results are discussed in relation to the survival value of the aggregations formed by these species; the validity of the term ‘resistantstage’ as applied to the 4th-stage larva of D. dipsaci is assessed.

There are about seven species of the genus Ascodipteron in the tropical parts of the Old World. Their endoparasitic females, which live embedded in the skin of bats, are not uncommon on certain species of the genera Emballonura, Hipposideros and especially of Rhinolophus and Miniopterus. The males, on the other hand, are extremely rare, and have never been taken on bats before. There is a description of the male of Ascodipteron speiserianum only, by Muir (1912), who bred this species in Amboina, Dutch East Indies.

1. The following new species were encountered in this investigation:

Haemogregarina hamata in Lacerta jacksoni;

Hepatozoon minchini in Crotaphopeltis degeni

Pirhaemocyton granosa in Agama colonorum

Atoxoplasma argyae n.gen. n.sp. in Argya rubi-ginosa

Spirochaeta harveyi in Cercopithecus aethiops.

2. Further information is given in regard to the following known species, including new host records:

Plasmodium pitmani in Mabuia striata and M. maculilabris

Trypanosoma martini in Mabuia maculilabris

Plasmodium vaughani in Othyphantes reichenowi

Plasmodium relictum in Othyphantes reichenowi and Passer iagoensis

Haemoproteus wenyoni in Passer griseus

Leucocytozoon sp. in nine species of birds

Hepatocystes epomophori in Epomophorus labiatus

Hepatocystes kochi in Cercopithecus, etc.

Babesia pitheci in Cercopithecus mitis.

3. Exo-erythrocytic schizogony is described in Plasmodium pitmani of the skink (from spleen and heart preparations).

Confocal scanning laser microscopy has been employed with immunocytochemical techniques to map the distribution of serotoninergic and peptidergic components in the nervous system of the monogenean gill-parasite, Diclidophora merlangi; results are compared with the distribution of cholinergic components, following histochemical staining for cholinesterase activity. While all three neurochemical elements are present in the central and peripheral nervous systems, the cholinergic and peptidergic systems dominate the CNS, whereas the PNS has a majority of serotoninergic nerve fibres. The cholinergic and peptidergic neuronal pathways overlap extensively in staining patterns, suggesting possible co-localization of acetylcholine and neuropeptides. Within the peptidergic nervous system, immunoreactivity to the pancreatic polypeptide family of peptides and FMRFamide were the most prevalent.

In dibacterial cultures of E. invadens the size of the cysts was significantly influenced by different species of concomitant bacteria, and by the composition of the media. These factors appear to influence cyst size as a result of their effect on the growth rate of the amoebae; a higher multiplication rate was associated with a decrease in the size of the cysts.

I wish to thank Dr A. Bishop for criticizing the manuscript, and Mr W. McConnachie, jun., for assistance with the statistical methods. The work has been financed by the Medical Research Council.

We have isolated a cDNA (SKvl. 1) encoding a Shaker-related K+ channel from an adult cDNA library of the human parasitic trematode Schistosoma mansoni. The deduced amino acid sequence (512 aa, 56·5 kDa) contains 6 putative membrane-spanning domains (S1–S6) and a pore-forming domain (H5). SKv1.1 is grouped in the Shaker family, but forms a unique branch within this family, on the basis of dendrogram analysis. SKv1.1 shows significant sequence identity with most other Shaker channels, with 64—74% identity in the core region (S1–S6). It has the highest sequence identity with the K+ channel (Ak01a) from Aplysia. Northern blot analysis detected a single primary transcript of 2·8 kb. Southern blot analysis indicated that SKv1.1 is present as a single copy in the genomic DNA of S. mansoni. Expression of SKv1. 1 in Xenopus oocytes produced a rapidly activating and inactivating outward K+ current which is highly sensitive to 4-aminopyridine, but is insensitive to tetraethylammonium, mast cell degranulating peptide, dendrotoxin and charybdo-toxin. The presence of a Shaker homologue in Schistosoma suggests that Sh subfamilies may exist in other lower invertebrates as well as platyhelminths.

The uptake of cycloleucine, L-proline, L-alanine and L-threonine by secondary hydatid cysts of Echinococcus granulosus (U.K. horse strain 3–8 mm in diameter, derived from Balb/c mice infected 300–400 days previously) occurs by passive diffusion into the cyst wall (laminated layer plus germinal layer) and by mediated mechanisms into the fluid-filled interior. The maximal concentrations of these compounds are achieved after incubation for 2 h in vitro and approach those in vivo. Kt and Vmax values describing the uptake of these compounds are given. The flux rates for these compounds are extremely slow compared to those obtained with the protoscolex. A rationale for standardizing the experimental method for uptake studies with hydatid cysts is described.

The freeze fracture technique has been used to study the internal cyto-architecture of the surface membranes of the parasite and erythrocyte in Plasmodium knowlesi infections. Six fracture faces, derived from the plasma membrane and 2 pellicular membranes, have been identified at the surface of the free merozoite. The apposed leaflets of the 2 pellicular membranes show the characteristic features of E fracture faces, a result compatible with the view that the pellicular membranes line a potential cisterna. There is evidence to suggest that there may be changes in the distribution and density of the integral proteins in the merozoite plasma membrane at invasion. Furthermore, vesicles consisting of stacked membranes occur within and around the erythrocyte invagination at invasion; it is suggested that these vesicles are released from the merozoite rhoptries. Formation of the parasitophorous vacuole is accompanied by dramatic changes in the density and distribution of intra-membraneous particles (IMP) in the vacuolar membrane. Initially there is a great reduction in particle numbers, but subsequently the particles reappear and show reversed polarity. The possible causes and implications of these changes are discussed. The intra-erythrocytic parasite synthesizes new transmembrane proteins as development proceeds, and the trophozoite and schizont stages of development are characterized by the appearance of circular, particle-free regions in the parasite plasmalemma. There is a decrease in the density of transmembrane proteins in the erythrocyte plasma membrane during parasite maturation, and the P face IMP show the characteristic features of aggregation.

The behaviour of cercariae was recorded on video at 10, 20 and 30 °C and a combination of population and individual behavioural patterns studied. Initially cercarial populations were extremely active, individuals alternating between swimming and sinking behaviour. As they aged the percentage sinking and resting increased resulting in the establishment of a vertical distribution in the water column. An increased environmental temperature reduced the active life-span of the parasites. Over time cercarial swimming rates declined. Swimming rates increased with environmental temperature and as a result of a shadow stimulus. As cercariae aged, the duration of swimming events decreased. Post-stimulation swimming periods were significantly longer than pre-stimulation periods. Pre- and post-stimulation swimming distances also declined with age. Post-stimulation swimming distances were significantly longer than pre-stimulation distances. The sinking rates of cercariae decreased with age while sinking periods and distances increased. Data for proportional time spent swimming and the ratio of sinking to swimming events indicated that as cercariae aged, less time was spent swimming. The frequency of swimming/sinking events declined with age. The implications of this behaviour for dispersal and transmission are discussed.

Naive CBA/Ca mice and CBA/Ca mice infected 12 weeks previously with 20 normal cercariae of Schistosoma mansoni were challenged percutaneously with isotopically labelled parasites. Challenge worm migration was followed, in 5 separate experiments, by means of compressed organ autoradiography. In three experiments, 43.1 % of challenge parasites did not arrive in the lungs of infected mice on day 6 as compared to 7.7 % in naive controls, thereby indicating that pre-lung sites constitute the first barrier in resistance to reinfection. A further 15% of the challenge worm burden was lost in the lungs or en route to the liver in the immune animals, and portal perfusion revealed that 25.4% of the challenge was lost in the liver. Two other experiments revealed no comparable phases of pre-liver attrition however; instead resistance was only evident at final perfusion on days 28 or 35. These data reveal the variable generation of specific acquired immunity in mice harbouring a chronic schistosome infection and thus clarify current discrepancies in the literature. The results are discussed in relation to documented evidence for the nature of specific and non-specific immune mechanisms reported to operate at different sites in the infection model of schistosomiasis mansoni.

The relationship of survival and reproduction of Gyrodactylus salaris Malmberg on the Atlantic salmon (Salmo salar) to water temperature (2·5–19·0 °C), was studied on the basis of temporal sequence of births and age at death of individual parasites on isolated salmon, and of infrapopulation growth on isolated and grouped salmon. Mean life-span of the parasite was negatively correlated with water temperature: 33·7 days at 2·5 °C and 4·5 days at 19·0 °C. The average number of offspring per parasite peaked between 6·5 and 13·0 °C, and was approximately 2·4 at these two temperatures. Both the period between the successive births of the offspring (max 4) and the estimated generation time were negatively correlated with temperature. The innate capacity for increase (rm) was positively correlated with temperature: from 0·02 (/parasite/day) at 2·5 °C to 0·22 (/parasite/day) at 19·0 °C. Growth of the infrapopulations was positively correlated with water temperature and was higher on isolated fish than on grouped fish, though less than the potential parasite population growth estimated from rm. In the infrapopulations the mean intensity of parasites continued to increase throughout all the experiments on both isolated fish and on grouped fish.

It has been shown in an earlier paper that if the eggs of parasites are not deposited in unparasitised hosts alone, but distributed according to the laws of chance among unparasitised hosts and hosts already attacked by individuals of the same species, and if all the parasites develop successfully, no matter how many enter a single host, then the effect upon the time required for the extermination of the host, as compared to that which is required if only unparasitised hosts are chosen, is relatively slight.

The hypothesis examined in this paper is that when several parasites enter a single host, one, and only one, will develop successfully. It is shown that in such a case, provided the reproductive rate is decidedly greater than that of the host (e.g. 1·5 times as great), the effect on the time required for the extermination of the host by the parasite will be very little if at all greater than when superparasitism causes merely a loss in efficiency, as in the hypothesis previously examined. When the reproductive rate of the host is practically or actually equal to that of the parasite, the process of control will be seriously retarded, the delay being very much greater than in the preceding hypothesis. Control will, however, occur, even in this case.

Superparasitism of the type here examined will therefore not necessarily produce any marked retardation of the process of control in cases where control within reasonable time would otherwise be probable.