The motility assay is the current gold standard for evaluating drug effects on hookworm larvae and adults, however, among other drawbacks the assay is time consuming, and prone to individual subjectivity. We evaluated six alternative in vitro assays, namely the feeding inhibition assay, the colourimetric AlamarBlue®, MTT formazan and acid phosphatase activity assays, as well as isothermal calorimetry and the xCELLigence System using Ancylostoma ceylanicum third-stage larvae, stimulated third-stage larvae and adults. The performances of the assays were compared to the motility assay using three standard drugs: albendazole, levamisole and ivermectin (100–1 μg/ml). None of the assays investigated offered an advantage over the motility assay, because they were all inapplicable to third-stage larvae, which were presumably metabolically and physically too inactive. Among all assays tested the xCELLigence System performed best on adult worms as the test was accurate, simple, required a minimal number of worms and offered the possibility for conducting a medium-throughput screening.

Class Rudimicrosporea Sprague 1977, with its single family Metchnikovellidae, comprises hyperparasites of gregarines from the guts of marine invertebrates. Metchnikovellids remain poorly studied in spite of their significance to the evolutionary history of microsporidia; their ultrastructure and life cycles require further investigation. Here we present results of the light- and electron-microscopy study of Metchnikovella incurvata Caulleri and Mesnil 1914, isolated from lecudinid gregarines, parasitizing polychaetes Pygospio elegans in the White Sea littoral zone, and yet described only on the light-microscopic level. The life cycle of this microsporidium includes 2 sporogonies: free (FS) and sac-bound (SBS). In FS, sporonts develop into multinuclear cells (sporogonial plasmodia), which generate sporoblasts and free spores residing in direct contact with the host cytoplasm. Electron microscopy revealed their metchnikovellidean structure: a horseshoe-shaped nucleus, short manubrium perpendicular to the long axis of the spore, and a polar cap in a separate membrane container. Merogony was not observed. The earliest stages of SBS were chains of binucleate cells. They underwent a series of nuclear and cell divisions, produced extracellular envelopes, and split into boomerang-shaped spore sacs, containing up to 16 spores each. Ultrastructure and sizes of sac-bounded spores were similar to those of free-living ones. An amended diagnosis of M. incurvata is provided.

Heparin-binding proteins (HBPs) have been demonstrated in both infective forms of Trypanosoma cruzi and are involved in the recognition and invasion of mammalian cells. In this study, we evaluated the potential biological function of these proteins during the parasite-vector interaction. HBPs, with molecular masses of 65·8 kDa and 59 kDa, were isolated from epimastigotes by heparin affinity chromatography and identified by biotin-conjugated sulfated glycosaminoglycans (GAGs). Surface plasmon resonance biosensor analysis demonstrated stable receptor-ligand binding based on the association and dissociation values. Pre-incubation of epimastigotes with GAGs led to an inhibition of parasite binding to immobilized heparin. Competition assays were performed to evaluate the role of the HBP-GAG interaction in the recognition and adhesion of epimastigotes to midgut epithelial cells of Rhodnius prolixus. Epithelial cells pre-incubated with HBPs yielded a 3·8-fold inhibition in the adhesion of epimastigotes. The pre-treatment of epimastigotes with heparin, heparan sulfate and chondroitin sulfate significantly inhibited parasite adhesion to midgut epithelial cells, which was confirmed by scanning electron microscopy. We provide evidence that heparin-binding proteins are found on the surface of T. cruzi epimastigotes and demonstrate their key role in the recognition of sulfated GAGs on the surface of midgut epithelial cells of the insect vector.

The nematode parasite, Nippostrongylus brasiliensis, induces a biphasic anorexia in its rat host. The mechanisms, underlying this anorexia and its possible advantages to the host or parasite are unknown. We have investigated the effect of acute (12–24 h) and chronic (2–17 days) infections on plasma concentrations of leptin, insulin and corticosterone, and on hypothalamic expression of neuropeptide Y, galanin and corticotrophin-releasing factor genes. Plasma leptin was elevated in infected rats relative to uninfected ad libitum-fed controls and pair-fed controls in 12 h infections initiated at dark onset and in infections of 2 days' duration. At other times prior to parasite expulsion, plasma leptin in infected and pair-fed rats was lower than that of uninfected ad libitum-fed controls, reflecting the existing state of negative energy balance. Elevated plasma leptin concentrations in infected rats at day 2 post-infection were accompanied by reduced neuropeptide Y gene expression in the hypothalamic arcuate nucleus compared with both ad libitum control and pair-fed animals, and by lowered corticotrophin-releasing factor gene expression in the paraventricular nucleus relative to pair-feds. Twelve hour infections were characterized by a substantial increase in plasma corticosterone that was independent of reduced food intake, and in 12 h infections initiated at dark onset, where plasma leptin was elevated, there was also increased plasma insulin concentration in infected rats. In longer infections, differences between the groups in plasma insulin and corticosterone concentration were only observed at day 4 post-infection. In summary, perturbations to leptin, insulin and corticosterone signals early in infection may have a causative role and might feed back onto hypothalamic gene expression, whereas subsequent changes in these parameters are more likely to be secondary to negative energy balance.

The effect of insect vectors on avian exposure to infection by pathogens remains poorly studied. Here, we used an insect repellent treatment to reduce the number of blood-sucking flying insects in blue tit Cyanistes caeruleus nests and examined its effect on nestling health status measured as body mass, nestling phytohaemagglutinin (PHA) response and blood parasite prevalence. We found that (i) the insect repellent treatment significantly reduced the number of blood-sucking flying insects in nests and (ii) the number of blood-sucking flying insects had a significant effect on the prevalence of the blood parasite Trypanosoma independently of the treatment. In addition, we found support for an adverse effect of parasite infections on nestling PHA response. Nestlings infected by Trypanosoma mounted a weaker response against PHA than non-parasitized ones. In addition, the number of blowflies in the nest was negatively associated with nestling PHA response. Overall, we found support for the hypothesis that blood-sucking flying insects attacking nestlings increase their exposure to parasite infections. Our results further substantiate the adverse effect of parasites on nestling condition.

Parasites are known to affect the predatory behaviour or diet of their hosts. In relation to biological invasions, parasites may significantly influence the invasiveness of the host population and/or mediate the relationships between the invader and the invaded community. Dikerogammarus villosus, a recently introduced species, has had a major impact in European rivers. Notably, its high position in trophic web and high predatory behaviour, have both facilitated its invasive success, and affected other macroinvertebrate taxa in colonized habitats. The intracellular parasite Cucumispora dikerogammari, specific to D. villosus, has successfully dispersed together with this amphipod. Data presented here have shown that D. villosus infected by this parasite have a reduced predatory behaviour compared with healthy individuals, and are much more active suggesting that the co-invasive parasite may diminish the predatory pressure of D. villosus on newly colonized communities.

The ultrastructure and development of the body wall of Polymorphus minutus is described during the acanthella stages in the intermediate host Gammarus pulex.

The cortex of the early acanthella is simple in structure, consisting of a syncytium containing giant nuclei, mitochondria, Golgi bodies, endoplasmic reticulum and a few lipid bodies. The cuticle is penetrated by pores which open into vesicles in a vesicular region.

The cortex gives rise to the body wall, neck and lemnisci of the adult, and may be considered as divided initially into two and later three functionally different regions: an outer absorptive region, an inner storage region and a layer between which develops to provide firm, resistant skeletal protection.

Absorption is facilitated by the increase in surface area due to the pores which penetrate the surface. This region is functional throughout development and the pores increase in number, possibly by division, as the animal grows. Beneath the pores a pore canal layer develops towards the end of the late acanthella stage.

During development supporting fibres are deposited throughout the cortex. Beneath the vesicular layer a concentration of mitochondria, Golgi bodies and endoplasmic reticulum deposit the fibres of the felt layer. In the anterior meta-soma no felt layer is found, but the body spines originate from this region.

The inner region of the cortex, or radial layer, contains the giant nuclei, which are arranged in the early acanthella in 5 rings of 4, 6, 5 or 6, 6 and 5 nuclei respectively. During development they produce dendritic process and finally fragment. They are considered here to be concerned with the formation of the felt layer and lemnisci rather than the ‘lacunar channels’. The inner cortical region becomes ensely packed with lipid bodies, and it is suggested that these may be responsible, owing to fixation artifact, for the so-called ‘lacunar channels’ previously described in these regions, the extensive cavities seen in some light microscope preparations being filled with a fluid lipid rather than an aqueous solution.

The lemnisci are similar in appearance to the cortex except for regions of concentrated endoplasmic reticulum.

The mitochondria in the cortex of the acanthella appear to be aerobic rather than anaerobic, as they possess many cristae and are larger than the mitochondria of the adults.

My thanks are due to Dr D. W. T. Crompton for advice and encouragement, and to Dr D. H. Northcote and Dr F. B. P. Wooding for time on the Phillips Electron Microscope 100. This work was carried out during the tenure of an S.R.C. studentship.

The in vitro absorption of glucose and l-methionine in the intestine of broiler chickens was measured 7, 14 and 21 days post-inoculation (p.i.) with sporulated oocysts of Eimeria acervulina, E. mivati, E. maxima or E. brunetti. The small intestine of each bird was divided into 8 regions of equal length and absorption was measured on 3 tissue disks of equal size from each region. The absorption rate of each substrate with each coccidial species was measured based on (1) an equal area from each region, (2) an equal weight from each region, (3) the total absorption in each region and (4) the total potential absorption in the intestine. Comparisons of absorption rate of equal areas in each intestinal region demonstrated that infected birds at 7 days p.i. absorbed significantly less substrate per unit area in the regions of maximum infection than uninfected controls. Malabsorption was less apparent when the weight of the region was used as the unit of measurement. Compensatory absorption was seen in some uninfected regions with E. acervulina. The total potential intestinal absorption at 7 days p.i. was reduced with E. mivati, E. maxima and E. brunetti but not with E. acervulina. At 14 days p.i., total l-methionine and glucose absorption in some regions of the intestine was significantly increased with E. acervulina but not with E. mivati, E. maxima or E. brunetti. No absorption differences were seen at 21 days p.i. with any species.

Iron-59 (as a ferric chloride solution or as a component of haemoglobin) was introduced into the lumen of the gut of Fasciola hepatica. It was detected by autoradiography in high concentrations in the intestinal epithelium, the cuticular epithelium, the myoblasts and the cells forming the walls of the excretory tubules. Carbon-14, introduced as l-phenylalanine-C-14, was concentrated in the same tissues, with the exception of the intestinal epithelium. These findings are discussed in relation to the function of the respective tissues.

Strains of mice poorly (B10) or non-responsive (B10.BR) to a primary infection with Trichuris muris were protected against infection by vaccination with excretory/secretory (E/S) antigen in Complete Freund's Adjuvant (CFA). Protection in these mice was slow to be expressed compared to that in good responder strains. Vaccination boosted the IgG and IgG1 antibody responses to E/S antigen and altered the antigen recognition profiles, three high molecular weight antigens (80–85, 90–95, 105–110 kDa) being recognized by antibodies in sera from vaccinated but not control mice. B10. BR mice which had experienced a patent primary infection could not be protected against challenge infections by vaccination and this was correlated with depressed levels of IgG1, but not total IgG, to E'S antigen early post-challenge compared with vaccinated infected mice which had not seen an adult primary infection. There was also lack of recognition of the three high molecular weight antigens recognized by antibodies in sera from mice infected after vaccination. It is suggested that the rapid development of high levels of IgG1 antibodies, and the recognition of the three high molecular weight antigens, may reflect events that are important in protective immunity. Immunomodulation of host immunity by T. muris may therefore be achieved, at least in part, by the suppression of specific IgG1 levels, the production of an irrelevant IgG isotype and prevention of the recognition of critical antigens.

Light and transmission electron microscopy were used to study different stages of Perkinsus atlanticus (Apicomplexa) during induced zoosporulation, with fluid thioglycollate medium and seawater. Cytokinesis and nucleokinesis of different developmental stages were studied every 12 h during the incubation period of 72 h, at which time the zoospores became free. Uninucleated and flagellated zoospores present the apical complex formed by conoid, polar ring, micronemes, rhoptries and subpellicular microtubules observed at different sections. Ultrastructural details were compared with the other two species of the genus Perkinsus.

A herd of 20 cattle was introduced on a ranch in Kilifi, Coast Province of Kenya, where they were in contact with Glossina austeni for 6 months. In total, 65 trypanosome isolates were made from these animals. Examination of the isolates revealed that 61 were Trypanosoma congolense and 4 were T. theileri. Out of the 61 T. congolense isolates, 55 were successfully passaged and cloned in mice to provide trypanosome populations for further analyses. The stocks and their clones were inoculated into goats on which teneral G. morsitans centralis were later fed in order to provide metacyclics for use in serodeme analysis. Identification of serodemes was carried out by indirect immunofluorescence and neutralization using antimetacyclic hyperimmune sera prepared in mice against metacyclics of cloned trypanosome populations. So far 4 serodemes have been identified in 8 stocks and 7 clones. Each of the 9 stocks contained a mixture of at least 2 of the 4 serodemes identified. Furthermore, stocks isolated sequentially from individual animals contained the same serodemes despite repeated treatment with a curative dose (6 mg/kg body weight) of Berenil between isolations. From the latter finding, it can be inferred that the 4 serodemes were present on the ranch throughout the study period.

Experiments are described on the inactivation by gamma-rays, X-rays, and alpha-rays of the viruses of tomato bushy stunt, tobacco necrosis, tobacco ringspot, tobacco mosaic and potato virus X. Within the errors of the experiment the inactivation curves appear to be exponential, and the inactivation doses increase in the order gamma-rays, X-rays, of wave-length 1·5 A., X-rays of wave-length 8·3 A., and alpha-rays.

A theory is given explaining these results and correlating the inactivation dose with the virus size. Estimates of the sizes of the viruses obtained from the radiation experiments he within the range of the sizes given by other methods, but are somewhat lower than the most probable sizes. Possible explanations of the discrepancy which are discussed are (a) the virus particle is not the molecule, in the sense of the smallest infective unit, or (b) certain structural changes in the virus molecule produced by the radiation may still leave it infective. Some of these may perhaps show themselves as mutations.

While I was in Egypt conducting investigations into the etiology of dysentery with. Captain F. W. O'Connor at Alexandria in 1916 I was able to make some observations on the intestinal protozoa of three species of lizard. The occurrence of an Entamoeba and a Chilomastix (Tetramitus) in one of these, Agama stellio, was mentioned in our publication on the Human Intestinal Protozoa in the Near East, p. 147 (1917). The other lizards investigated were Chamaeleon vulgaris and Lacerta agilis. I will describe the observations on the protozoa under their respective hosts.

On three occasions, antibody positive blood from wild red deer produced overt infections with Babesia when inoculated into splenecto-mized red deer. One of the deer also became infected with Eperythrozoon sp. Babesia divergens, B. capreoli and the Babesia of red deer are morphologically similar and the marginal position of the parasites in the host cell is characteristic. Babesia were not seen and no antibody was formed in five out of six splenectomized bovine calves which were injected with parasitaemic red deer blood. Two of these calves when challenged with B. divergens were fully susceptible. A transient infection with the deer Babesia may have occurred in the sixth calf since antibody was detected and the animal resisted challenge with B. divergens.In indirect fluorescent antibody tests there was little or no difference in the titre of sera from naturally or experimentally infected cattle and deer when reacted with B. divergens or the red deer Babesia antigens. Despite their similarities, specific status for B. divergens and the red deer Babesia is probably justified; at present there is insufficient evidence to justify separation of the red deer Babesia from B. capreoli.

1. The life-history of Phaenoserphus viator is described.

Four larval instars are found, endoparasitic in the larvae of Pterostichus niger. At thee nd of the last larval instar the parasites, which may number as many as 45 in a single host, emerge, and while still attached, pupate without spinning a cocoon.

Adults may appear in August or September.

The effect of the parasite in inhibiting metamorphosis of the host is discussed.

2. The first observed larva is atracheate and incompletely segmented at first and is of the polypod type bearing paired prolegs on the body segments.

Subsequent instars are apodate.

The tracheal system develops progressively in the several instars, but only becomes functional in the final stage.

3. The anatomy of the larva is briefly described with the exception of the musculature.

Tracheal development is described. Gas only appears in the tracheae after the development of the tracheole cells puts the tracheae into communication with the body wall and other organs.

In the circulatory system an important accessory organ is the neural sinus, formed by the enclosure of the ventral nerve cord beneath a connective tissue curtain.

The imaginal discs of the hypodermis are briefly described, these being clearly defined in the head, thorax, and posterior abdominal segments.

The nervous system consists of a brain, suboesophageal ganglion and 11 ventral ganglia, the most posterior being tripartite. This system is connected with the sympathetic, by nerves passing from the cerebral commissures to a frontal ganglion which lies above the oesophagus and behind the labrum.

The reduction in worm burden obtained by treatment of Schistosoma mansoni with praziquantel and oxamniquine was greater in mice with heavy infections than in relatively lightly infected animals. The reduction in worm burden achieved by each drug correlated with the size of the pre-treatment worm burden (r2 = 0·82 and 0·81 for praziquantel and oxamniquine, respectively). Intensity of infection did not affect the degree of tegumental damage and drug-induced antigen exposure on worms recovered soon after treatment with praziquantel. However, praziquantel-treated worms from mice with heavy infections had significantly more murine antibody attached to the treated-worm surface than worms from praziquantel-treated lightly infected mice. Heavily infected mice had greater levels of circulating anti-worm antibodies than lighter infected mice. The correlation between infection intensity and cure rates achieved by praziquantel and oxamniquine may thus be a reflection of the higher litres of relevant antibody in heavily infected mice mediating death of drug-treated worms.

Since its use as a model to study metazoan parasite culture and in vitro development, the plerocercoid of the tapeworm, Ligula intestinalis, has served as a useful scientific tool to study a range of biological factors, particularly within its fish intermediate host. From the extensive long-term ecological studies on the interactions between the parasite and cyprinid hosts, to the recent advances made using molecular technology on parasite diversity and speciation, studies on the parasite have, over the last 60 years, led to significant advances in knowledge on host-parasite interactions. The parasite has served as a useful model to study pollution, immunology and parasite ecology and genetics, as well has being the archetypal endocrine disruptor.

This study examined whether Ascaridia galli infection can be controlled by dietary non-starch polysaccharides (NSP) in chickens. One-day-old chicks were fed either a basal diet (CON) or CON plus insoluble NSP (I-NSP), or CON plus soluble NSP (S-NSP) for 11 weeks. Three weeks later, birds from half of each feeding group were inoculated with 250 embryonated eggs of A. galli, and slaughtered 8 weeks post-infection to determine worm counts. Both NSP diets, particularly S-NSP, increased prevalence of infection (P<0·05) and worm burden (roughly +50%) of the birds (P<0·001). A. galli infection caused a less efficient (P=0·013) feed utilization for body weight gain (BWG) resulting in lower body weights (P<0·001) irrespective of type of diet consumed. NSP-fed birds, particularly those on I-NSP, consumed more (+8%) feed per unit BWG and showed retarded (P<0·001) BW development compared to CON-fed birds. Intracaecal pH was lowered by S-NSP (P<0·05). Both NSP diets increased the volatile fatty acids pool size in caeca (P<0·001) with S-NSP exerting a greater effect (+46%) than I-NSP (+24%). It is concluded that both NSPs supplemented diets alter gastrointestinal environment in favour of the nematode establishment, and thus have no potential for controlling A. galli infection in chickens.

Studies of the use of parasites as biological tags for stock identification and to follow migrations of marine fish, mammals and invertebrates in European Atlantic waters are critically reviewed and evaluated. The region covered includes the North, Baltic, Barents and White Seas plus Icelandic waters, but excludes the Mediterranean and Black Seas. Each fish species or ecological group of species is treated separately. More parasite tag studies have been carried out on Atlantic herring Clupea harengus than on any other species, while cod Gadus morhua have also been the subject of many studies. Other species that have been the subjects of more than one study are: blue whiting Micromesistius poutassou, whiting Merlangius merlangus, haddock Melanogrammus aeglefinus, Norway pout Trisopterus esmarkii, horse mackerel Trachurus trachurus and mackerel Scomber scombrus. Other species are dealt with under the general headings redfishes, flatfish, tunas, anadromous fish, elasmobranchs, marine mammals and invertebrates. A final section highlights how parasites can be, and have been, misused as biological tags, and how this can be avoided. It also reviews recent developments in methodology and parasite genetics, considers the potential effects of climate change on the distributions of both hosts and parasites, and suggests host-parasite systems that should reward further research.