The glycosyl-phosphatidylinositol (GPI) protein-membrane anchors are ubiquitous among the eukaryotes. However, while mammalian cells typically express in the order of 100 thousand copies of GPI-anchor per cell, the parasitic protozoa, particularly the kinetoplastids, express up to 10–20 million copies of GPI-anchor and/or GPI-related glycolipids per cell. Thus GPI-family members dominate the cell surface molecular architecture of these organisms. In several cases, GPI-anchored proteins, such as the variant surface glycoprotein (VSG) of the African trypanosomes, or GPI-related glycolipids, such as the lipophosphoglycan (LPG) of the Leishmania, are known to be essential for parasite survival and infectivity. The highly elevated levels and specialised nature of GPI metabolism in the kinetoplastid parasites suggest that the GPI biosynthetic pathways might be good targets for the development of chemotherapeutic agents. This article introduces the range of GPI structures found in protozoan parasites, and their mammalian hosts, and discusses some aspects of GPI biosynthesis.

Muscle fibres isolated from adult Schistosoma mansoni contracted in a dose-dependent manner when exposed to elevated K+ with a maximum response obtained with 25 mM K+. These contractions were dependent on extracellular Ca2+ since Co2+ (5 mM) or nicardipine (1 μM) blocked the high K+ contractions. Serotonin (300 nM or higher) was required for maintenance of high K+ contractions. With concentrations of serotonin less than 300 nM the response was dose dependent. 5-Methoxytryptamine or α-methylserotonin at 1 μM as well as 10 μM tryptamine were able to substitute for serotonin, but 1 μM 5-carboxyamidotryptamine was ineffective. The order of potency for antagonists (10 μM) was: methiothepin > metergoline > Ly-278,584 = ketanserin. This pattern of responsiveness does not fit well with any known mammalian serotonin receptor subtype. Since forskolin, an adenylate cyclase activator, is able to mimic the action of serotonin and H89, a protein kinase inhibitor, is able to block the effect of serotonin, the effect of serotonin on contractility of the muscle may be via a cAMP-dependent pathway.

The developmental expression of the antigenic fucosylated glycan motifs Fucα1-3GalNAcβ1-4GlcNAc (F-LDN), Fucα1-3GalNAcβ1-4(Fucα1-3)GlcNAc (F-LDN-F), GalNAcβ1-4(Fucα1-3)GlcNAc (LDN-F), Galβ1-4(Fucα1-3)GlcNAc (Lewis X), and GalNAcβ1-4(Fucα1-2Fucα1-3)GlcNAc (LDN-DF) in Schistosoma mansoni cercariae, adult worms and eggs, was surveyed using previously defined anti-carbohydrate monoclonal antibodies (mAbs). Lewis X was found both on glycolipids and glycoproteins, yet with completely different expression patterns during the life-cycle: on glycolipids, Lewis X was mainly found in the cercarial stage, while protein-conjugated Lewis X was mainly present in the egg stage. Also protein-conjugated LDN-F and LDN-DF were most highly expressed in the egg-stage. On glycolipids LDN-DF was found in all three examined stages, whereas LDN-F containing glycolipids were restricted to adult worms and eggs. The motifs F-LDN and F-LDN-F were found both on glycoproteins and glycolipids of the cercarial and egg stage, while in the adult stage, they appeared to occur predominantly on glycolipids. Immunofluorescence assays (IFA) showed that these F-LDN and F-LDN-F containing glycolipids were localized in a yet undefined duct or excretory system of adult worms. Murine infection serum showed major reactivity with this adult worm duct-system, which could be fully inhibited by pre-incubation with keyhole limpet haemocyanin (KLH). Clearly, the use of defined mAbs provides a quick and convenient way to map expression profiles of carbohydrate epitopes.

Isolates of Cryptosporidium muris and C. serpentis were characterized from different hosts using nucleotide sequence analysis of the rDNA 18S and ITS1 regions, and the heat-shock (HSP-70) gene. Phylogenetic analysis confirmed preliminary evidence that C. muris is not a uniform species. Two distinct genotypes were identified within C. muris; (1) C. muris genotype A; comprising bovine and camel isolates of C. muris from different geographical locations, and (2) C. muris genotype B comprising C. muris isolates from mice, a hamster, a rock hyrax and a camel from the same enclosure. These 2 genotypes may represent separate species but further biological and molecular studies are required for confirmation.

The histology and ultrastructure of the salivary gland in Dermacentor andersoni are presented with particular emphasis on those aspects relating to fluid secretion. We suggest that the group III acinus contributes most of the fluid portion of the saliva (i.e. water and small molecules) and that the main cell-type involved is what we name the ‘water-cell’. The granule-cells possibly secrete the cement by which the tick secures its mouthparts to the host, and the ‘vacuolar cell’ possibly produces a protein-rich secretion. The function of the group I acinus remains obscure.

The genetic basis of virulence in a line (YM) of Plasmodium yoelii yoelii was investigated in a cross with a mild line (A/C). The blood forms of the virulent line developed extensively in mature erythrocytes of mice, causing death of the host within 7 days; infections with the mild line were normally restricted to reticulocytes, infected animals recovering after three weeks. Lines YM and A/C differed additionally in enzyme and drug-sensitivity markers. Studies on infections established from each line alone from sporozoite mixtures of the two lines and from the cross between the lines showed that the appearance of virulence had been caused by a genetic change in the parasite, and not by other factors such as a concurrent infection with another organism. An analysis of the characters of 56 clones derived from the cross showed that the virulence character had undergone recombination with the other markers, and appeared to be inherited in Mendelian fashion. Three clones exhibited atypical virulence, although it was not clear whether this had been produced by genetic recombination.

Classical studies of non-immune individuals infected with Plasmodium falciparum reveal that the infection may be regulated for long periods at a relatively stable parasite density, despite the enormous growth potential of a parasite that continually replicates within host erythrocytes. This suggests that the parasite population may be controlled by density-dependent mechanisms, and in theory the most obvious of these is competition between parasites for host erythrocytes. Here we evaluate the role of this mechanism in the regulation of parasitaemia, by modelling the basic population interaction between parasites and erythrocytes in a form that allows all the essential parameters to be estimated from clinical data. Our results show that competition cannot account for the total regulation of P. falciparum, but when combined with immune mechanisms it may play a more important role than is generally supposed. Further analysis of the model indicates that in the long term, parasite replication at low parasite densities can contribute significantly to the high degree of anaemia observed in natural infection, a conclusion which is not obvious from simple clinical observation.

An instrument for measuring the motility of larval and adult nematodes is described along with an analysis of its use as a tool to analyse drug action on these parasites. Motility was detected from larval parasites of Nippostrongylus brasiliensis and Ascaris suum in the absence and presence of various anti-nematodal drugs. These agents produced, within 48 h, a significant decrease in larval parasite motility. The instrument was also capable of detecting the motility of Caenorhabditis elegans, adult female Brugia pahangi and their response to anti-nematodal drugs. The design of the instrument allows us to accurately measure motility in a single sample within 60 sec.

A segment of the ribosomal RNA gene of Schistosoma mansoni and a DNA fragment specific to Echinococcus granulosus, cloned in plasmids, have been used as DNA probes to assess the extent of genetic variability within E. granulosus and some distinct strains have been identified. The DNA analysis, involving restriction endonuclease digestion and Southern blot hybridization with the probes, did not demonstrate any significant genetic variation within the U.K. horse/dog or sheep/dog strains but confirmed the distinctiveness of the two strains shown in previous studies. The sheep/dog strain was shown to be cosmopolitan in its distribution and fertile bovine material originating from the United Kingdom, Kenya, Spain and India conformed to this strain by DNA hybridization. In contrast, cattle isolates from Holland produced markedly different DNA hybridization banding profiles indicating that cattle can harbour more than one strain of E. granulosus. Similarly, it was shown that goats can harbour two different strains of E. granulosus, the sheep/dog strain and a form which infects camels. The strain of E. granulosus infecting equines in Spain and Ireland is genetically identical to that infecting horses in the United Kingdom. There is also a different strain infecting pigs in Poland and Yugoslavia. This pig/dog strain appears to be very similar genetically to the forms of E. granulosus which use camels and goats as intermediate hosts and is similar, though not identical, to the variant infecting Dutch cattle. It has been shown that E. granulosus material, fixed for a prolonged period in ethanol, or lyophilized, is amenable to DNA analysis and that it is possible to characterize the DNA of a single adult worm.

The neuropeptide story began in 1928 with the description by Ernst Scharrer of gland-like nerve cells in the hypothalamus of the minnow, Phoxinus laevis. Because these nerve cells were overwhelmingly specialized for secretory activity, overshadowing other neuronal properties, Scharrer termed them ‘neurosecretory neurons’. What was even more remarkable about the cells was that their products were released into the bloodstream to act as hormones, specifically neurohormones. Neurosecretory cells were identified largely on morphological grounds. That is, they could be stained with special techniques, such as chrome-haematoxylin and paraldehyde-fuchsin, although the techniques are far from specific, staining non-neurosecretory cells as well. However, the basis for the ‘special’ neurosecretory techniques is the demonstration of sulphur-containing proteins – so they are indicative of peptide-producing neurones. An alternative characteristic of neurosecretory cells is the presence of large (> 100 nm), dense-cored vesicles at the electron microscope level; these are the so-called elementary granules of neurosecretion, or ENGs. However, implicit in the concept of neurosecretion is that the prime function of the neurosecretory cell is in endocrine regulation, exerting a hormone-like control over some aspect of the organism's metabolism, by controlling endocrine glands and other effector organs. To satisfy this criterion, evidence had to be obtained of cycles of secretory activity within the cell that could be correlated with a change in the physiological condition of the organism.

Predator-prey models are often applied to the interactions between host immunity and parasite growth. A key component of these models is the immune system's functional response, the relationship between immune activity and parasite load. Typically, models assume a simple, linear functional response. However, based on the mechanistic interactions between parasites and immunity we argue that alternative forms are more likely, resulting in very different predictions, ranging from parasite exclusion to chronic infection. By extending this framework to consider multiple infections we show that combinations of parasites eliciting different functional responses greatly affect community stability. Indeed, some parasites may stabilize other species that would be unstable if infecting alone. Therefore hosts' immune systems may have adapted to tolerate certain parasites, rather than clear them and risk erratic parasite dynamics. We urge for more detailed empirical information relating immune activity to parasite load to enable better predictions of the dynamic consequences of immune-mediated interspecific interactions within parasite communities.

Cyathostomins are the most prevalent equine intestinal parasites and resistance has been reported in these nematodes against all 3 licensed anthelmintic classes. Strategies need to be developed that are less dependent upon drugs and more reliant on management-based control. To develop these we need to understand natural transmission patterns better. Here, we analysed longitudinal fecal egg count (FEC) data from 5 pony populations used for conservation purposes. We tested how egg excretion varied amongst populations and individuals, and how this was affected by age and climate. There was evidence for consistency in FECs over time at the individual level; this was generally weak and accounted for <10% of the total variance. Animals <5 years old had higher FECs and there was profound seasonal variation in FECs, with highest levels recorded in spring/summer. Effects of monthly temperature and rainfall explained most, but not all, of the observed seasonal variation and associations between climate measures and FECs were stronger in younger versus adult animals. One population was occasionally treated with anthelmintics and analysis of this population suggested that treatment substantially altered the seasonal dynamics. This paper highlights the variability in strongyle egg excretion amongst individuals and the factors involved in this variation.

A detailed description of the musculature in the different regions of the alimentary canal of Calliphora erythrocephala is given, and an account of the structure and function of the crop, proventriculus, ducts of the Malpighian tubes, rectal valve and rectal papillae. It has been shown by dissections and experiments that a system of channels exists in the rectal papillae through which the body fluid probably circulates, and it is suggested that the very large cells may have functions resembling those of liver cells.

The number of schistosomula recovered from abdominal skin declined exponentially with time after infection. However, this decline was shown to be an untrue reflection of the numbers present because of changes in recoverability on days subsequent to infection. Digestion of skin by the enzymes collagenase and hyaluronidase produced a small increase in the number of schistosomula recovered. The number of schistosomula present in abdominal skin and capable of reaching maturity was estimated by exciting the area to which cercariae had been applied, from groups of hosts at daily intervals after exposure. The number of worms maturing in each group of hosts was then determined. The mean duration of stay of schistosomula in mouse, hamster and rat abdominal skin was estimated at 88, 64·5 and 70 h, respectively. The fluid content of abdominal skin to which cercariae had been applied, increased following infection but it was concluded that this alone was not the predominant factor controlling exit of worms. The stimulation of a delayed hypersensitivity response not directed against schistosomes (in mice previously sensitized with 1-chloro,2–4-dinitrobenzene (DNB)) 1 day after exposure to cercariae, produced a significant increase in the number of schistosomula leaving the skin and reaching maturity. Schistosomula can be recovered from the axial lymph nodes of mice, hamsters and rats. If the schistosomula from mice are injected into the tail vein of uninfected mouse hosts, they have the same chance of maturing as comparable skin schistosomula. They are therefore apparently unharmed by their passage through the nodes. Interference with lymphatic drainage by surgical removal of lymph nodes has no effect on the number of schistosomula recovered from the lungs, but the peak recovery is delayed by 24 h. The hamster cheek-pouch, a site with no afferent lymphatic drainage, was also used to investigate the importance of the lymphatic exit route. There was no reduction in the number of adult worms recovered following infection at this site compared with the normal abdominal skin site. The migration of schistosomula to deeper tissues and their entry into blood vessels was followed using the cheek-pouch preparation. It was concluded that the major route of exit from the skin is the blood vascular system.

The course of infection with a local strain of Schistosoma mansoni was investigated in Kenyan baboons (Papio anubis). A single exposure to 5000 cercariae per baboon (cpb) killed five 4–8 kg baboons; a sixth survived the full 30-week post-exposure period, with suppressed egg production after week 11. Lower doses were not lethal. A single exposure to 1000 cpb gave satisfactory adult worm recoveries, high and steady faecal egg excretion, and a high tissue egg recovery, predominantly from the large intestine. A similar but less consistent picture was obtained with a single exposure to 200 cpb. Baboons receiving 5 monthly exposures of 200 cpb (1000 cpb total) showed some evidence of the development of protective immunity. Three animals showed patterns resembling those for 200 and 1000 cpb. The other 3 had suppressed egg production (faecal and tissue) indicating, perhaps, some immunological process. They also had an abnormally high percentage of eggs in the liver.

The morphology and life cycle of an amoeba producing amoebiasis in reptiles is described. The organism is assigned to the species Entamoeba invadens Rodham 1934, because of the pathogenicity of that species.

Material for the study was derived from 30 individuals of 7 species of reptiles, either dying of spontaneous amoebiasis or experimentally infected. (Material from 3 additional species of reptiles dying of amoebiasis was not used in this study.)

With the exception of a few minor morphological differences, E. invadens was found to be strikingly similar to E. histolytica in morphology of trophic and cystic stages, and in the processes of encystation, excystation, and metacystic development.

Metacystic development was studied in stained preparations made from the intestines of snakes 1–24 hours after the inoculation by mouth of mature cysts. Excystation usually takes place in the jejunum and ileum, a single quadrinucleate amoeba hatching from each cyst. Each quadrinucleate metacystic amoeba ultimately forms 8 amoebulae as a result of one division of each cystic nucleus, cytoplasmic division, and parcelling out of the cystic (N) and daughter (n) nuclei. Nineteen of the 24 possible combinations of N and n nuclei were observed.

The culture phenomena of multiple fission without encystment is suggested, and cannibalism of cysts by the trophozoites is described.

The pattern of recovery of schistosomula from the lungs of mice, hamsters and rats is described following normal percutaneous infection via the abdominal skin. Peak numbers were found in hamsters and rats on day 5 and in mice on day 6 post-exposure. Schistosomula were recovered from the lungs up to approximately day 20 post-exposure in all 3 species. None were found in pleural washings and only 2 were recovered following mincing and incubation of host diaphragms. The pattern of recovery of schistosomula from the lungs of mice was also described following the injection of a pulse of parasites into the tail vein. Approximately 54% of these injected schistosomula eventually reached maturity in the hepatic portal system. The ability of schistosomula to migrate and mature following their injection into unusual locations was tested. Small numbers reached maturity in the hepatic portal system following injection into the pleural cavity or subcutaneously, more when injected intra-peritoneally and largest numbers following intravenous injection. Small numbers of schistosomula were recovered by mincing and incubation of systemic organs such as the kidney and spleen. Schistosomula injected into the left ventricle of hamsters were able to migrate via the systemic organs to the lungs and 47% eventually matured in the hepatic portal system. The pattern of schistosomulum accumulation in the hepatic portal system of mice, hamsters and rats is described. Following normal percutaneous infection, schistosomula were first detected in this site from day 6 to day 9 depending on the host species. In mice, migration was complete around day 20 but continued in hamsters up to day 40 post-exposure. Following injection of schistosomula into the tail veins of mice, the first arrivals in the hepatic portal system were detected 12 h later and were found to lose their ability to migrate shortly after arrival. However, a proportion of lung schistosomula injected directly into the hepatic portal system were able to traverse hepatic sinusoids and reach the lungs. It was concluded that the route of migration of schistosomula from lungs to liver was entirely intravascular, with potentially several passages round the pulmonary-systemic circulation, before chance entry into arteries leading to the hepatic portal system. The proportion of schistosomula exiting from the pulmonary-systemic circuit was estimated as 0·14/day. A computer simulation produced values of 11 h and 5 h for the duration of migration through pulmonary and systemic capillary beds. The time for 1 complete circuit would thus be 16 h and the proportion of schistosomula exiting per circuit would be 0·095. This approximates to the proportion of cardiac output going to splanchnic organs in the resting rat (0·128). The narrowness of hepatic sinusoids may be one factor contributing to the sequestration of schistosomula in the hepatic portal system.

Different populations of the whitespotted rabbitfish, Siganus sutor, were examined for ectoparasites: adults from the Mombasa area (sampled in December 1990) and different age classes (adult, subadult and juveniles) from Gazi Bay (sampled in December 1992 and August 1993). The most common gill parasites were: the monogeneans Pseudohaliotrema sp., Tetrancistrum sigani and Microcotyle mouwoi, the copepods Hatschekia sp., Pseudolepeophtheirus sp. and juvenile Caligidae, and prazina larvae of the isopod Gnathia sp. Adult siganids had a higher parasite load than subadults. Juvenile rabbitfish did not harbour any gill parasites. Temporal differences in the parasite load of subadult rabbitfish were observed for M. mouwoi (highest in the December samples) and for juvenile Caligidae (highest in August). The microhabitat of the 5 most common gill parasites was species specific. Most parasite species showed distinct site preferences with respect to both gill arches and gill sectors, within the gill arches. Niche breadth of the different gill parasite species was independent of the abundance of any of the other species present. However, niche breadths of M. mouwoi, Tetrancistrum sp. and Hatschekia sp. increased with their own abundance. This suggests that interspecific competition for space is low and that intraspecific factors could play an important role in the microhabitat choice of these gill parasites. The hypothesis that niche restriction leads to higher intraspecific contact and an enhancement of chances to mate was tested on 2 monogenean species, Pseudohaliotrema sp. and Tetrancistrum sigani. Their highly aggregated distribution over the gill filaments, leading to increased intraspecific contact, is consistent with the hypothesis.

The genetic polymorphism among 271 Danish Cryptosporidium isolates of human and animal origin was studied by partial amplification and sequencing of the Cryptosporidium oocyst wall protein (COWP) gene, the 18S rDNA, and a microsatellite locus.The nucleotide sequence data reported in this paper are available in the GenBank database under the accession number AF469174. Furthermore, the microsatellite locus was studied directly using fragment analysis. A comparative analysis of DNA sequences showed the presence of 3 different subgenotypes (C1, C2 and C3) in C. parvum isolates from Danish cattle, with prevalences of 16.7, 17.2 and 73.1% including 13 (7.0%) mixed infections. Subgenotype C1 was significantly more prevalent (P<0.001) in the southern part of Denmark. In Cryptosporidium isolates of human origin the anthroponotic subgenotype H1 was identified, in addition to the zoonotic subgenotypes C1, C2, and C3. Of 44 human samples, 56.8% were anthroponotic, whereas 40.9% were zoonotic genotypes. One human isolate was characterized as C. meleagridis. The porcine Cryptosporidium isolates (N = 4) revealed a pattern which was genetically distinct from human and bovine isolates. Cryptosporidium in a hedgehog (Erinaceus europaeus L.) was identified for the first time. By microsatellite sequencing the hedgehog isolate showed a subgenotype distinct from the previously detected types. The assignment to subgenotype by microsatellite sequencing and fragment typing was 100% identical in samples where results were achieved by both methods. In addition, the fragment analysis proved more sensitive, easier, faster, and less expensive compared to sequencing.

DAMP (3-(2,4-dinitroanilino)-3′amino-N-methyldipropylamine), which differentially accumulates in acidic compartments, was used to identify such compartments in Toxoplasma gondii tachyzoites at the electron microscope level. In both free tachyzoites and dividing intracellular parasites the only sites of DAMP accumulation were mature and forming rhoptries. No labelling of other secretory organelles (micronemes and dense granules), the ER, Golgi or any other membrane-bounded organelles or anything resembling a lysosomal system was observed. Labelling of the forming rhoptries was higher and more homogenous than in mature rhoptries in which labelling was confined to the expanded ends of each organelle. The acid pH-dependent accumulation of DAMP in the forming and mature rhoptries was blocked by ammonium chloride and monensin, reagents known to abolish intracellular pH gradients. Estimates of rhoptry pH, based on the level of DAMP accumulation, show that the intralumenal pH of forming rhoptries is more acidic (pH 5·5–3·5) than the mature rhoptries (pH 7·0–5·0).